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- Nat Med 17(2):137 (2011)
Nature Medicine | Editorial The scientific social network Journal name:Nature MedicineVolume: 17,Page:137Year published:(2011)DOI:doi:10.1038/nm0211-137Published online04 February 2011 A joint statement from 17 funding agencies urges biomedical researchers to openly share data obtained from population-based studies. Although this will foster more collaboration, new web technologies need to be harnessed, and the attribution of credit must change to facilitate this transition. View full text Additional data - Are advance market commitments for drugs a real advance?
- Nat Med 17(2):139 (2011)
Nature Medicine | News Are advance market commitments for drugs a real advance? * Megan Scudellari Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:139Year published:(2011)DOI:doi:10.1038/nm0211-139Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. In December, health leaders in Nicaragua celebrated the integration of a vaccine against pneumococcal disease into their country's routine immunization program. It's no small achievement: pneumococcal disease kills more than 800,000 children under five years of age every year worldwide. What's perhaps more notable is that the change was made possible by an advance market commitment (AMC), a first-of-its-kind initiative devised by the GAVI Alliance in which donors provide funds to incentivize pharmaceutical companies to develop and distribute vaccines for the world's poorest nations. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Analysts see 'tale of two CEOs' unfolding at Pfizer and Merck
- Nat Med 17(2):140 (2011)
Nature Medicine | News Analysts see 'tale of two CEOs' unfolding at Pfizer and Merck * Stu Hutson Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:140Year published:(2011)DOI:doi:10.1038/nm0211-140Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. "It was the best of times, it was the worst of times..." Charles Dickens may not have lived during the age of blockbuster drugs such as Lipitor and Vioxx, but his writing captures the state of the pharmaceutical industry. Although product pipelines are drying up, the emergence of new markets, coupled with the advent of personalized medicine and other advances, offer exciting opportunities for drugmakers. And, given the unpredictable drug-development environment, analysts are keeping a watchful eye as a 'tale of two CEOs' unfolds at two top pharmaceutical firms. This year kicks off with new chief executives at two of the largest drugmakers, New Jersey–based Merck and Pfizer, headquartered in New York. One leadership change seems to show a company sticking to a carefully plotted course. The other, a more unexpected switch at the helm, reveals a pharmaceutical giant with a fluctuating business approach. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Action urged on foreign takeovers of Indian drugmakers
- Nat Med 17(2):141 (2011)
Nature Medicine | News Action urged on foreign takeovers of Indian drugmakers * T V Padma Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:141Year published:(2011)DOI:doi:10.1038/nm0211-141aPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. NEW DELHI — On the face of it, the takeover of six of Indian's key drug firms by major foreign players in the past four years seems to be routine business. But people within the government and industry watchdogs in India have started to worry. In 2008, Japan's Daiichi-Sankyo took control of India's largest drugmaker, Ranbaxy Laboratories, located about 20 miles south of New Delhi. Other Indian firms that have met a similar fate include Dabur Pharma, Shantha Biotech, Piramal Healthcare, Matrix Laboratories and Orchid Chemicals and Pharmaceuticals. Local concern grows out of the fact that these companies are major producers of cheap generic versions of essential medicines and vaccines, with wide market access in India and in other developing countries. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Transparency initiative moves ahead despite official's departure
- Nat Med 17(2):141 (2011)
Nature Medicine | News Transparency initiative moves ahead despite official's departure * Charlotte Schubert Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:141Year published:(2011)DOI:doi:10.1038/nm0211-141bPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. An initiative to increase transparency at the US Food and Drug Administration (FDA) moved forward on 6 January with the release of a report detailing steps the agency says it is undertaking to make its inner workings more evident to the drug industry. The report, FDA Transparency Initiative: Improving Transparency to Regulated Industry, includes 19 so-called 'action items' for implementation this year, ranging from posting presentations by agency employees online to responding faster to questions from industry. The document also lists five draft proposals—now up for public comment—to further improve transparency. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Compromise is in sight for new embryo research rules in France
- Nat Med 17(2):142 (2011)
Nature Medicine | News Compromise is in sight for new embryo research rules in France * Barbara Casassus Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:142Year published:(2011)DOI:doi:10.1038/nm0211-142aPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. PARIS — In 2004, France made a radical reversal by lifting its total ban on human embryonic stem cell research. But the law governing approval to conduct these types of studies remains convoluted, and scientists worry that the current system is dissuading companies from setting up research outfits in France. Proposals to update the French bioethics law will finally be presented to Parliament in early February, and a compromise may be on its way to help satisfy all partisans on this score. Scientists are virtually unanimous in the opinion that the current ban with exemptions should be dropped, but politicians remain divided over the question. "The opposition comes mainly from the Roman Catholics and crosses party lines," says Jean-Sébastien Vialatte, a parliamentarian and vice president of a special all-party National Assembly committee for the new law. Although France has been a staunchly secular country since 1905, "there has been strong pressure from the Church," he adds. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - New regulations urged for UK health research
- Nat Med 17(2):142 (2011)
Nature Medicine | News New regulations urged for UK health research * Bea Perks Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:142Year published:(2011)DOI:doi:10.1038/nm0211-142bPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Complex regulation and governance of clinical research in the UK has held back research with no evidence of improved patient safety, concludes a report by the country's Academy of Medical Sciences (AMS). The AMS was commissioned by the UK government in March 2010 to review the regulation of clinical and health research. The review, released on 10 January, recommends the establishment of an independent Health Research Agency (HRA) to simplify approval processes that currently differ across the UK. The authors also recommend the establishment of a National Research Governance Service within the HRA to accelerate approval of multicenter studies. The branch would take responsibility for certain approvals of multicenter trials; the current system requires multiple approvals carried out by different units within the country's National Health Service. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Animal instinct helps doctors ferret out disease
- Nat Med 17(2):143 (2011)
Nature Medicine | News Animal instinct helps doctors ferret out disease * Michelle Pflumm Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:143Year published:(2011)DOI:doi:10.1038/nm0211-143Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. In 2007, David Dosa, a geriatrician at Brown University in Providence, Rhode Island, reported the curious case of a white-bellied cat that purportedly could predict death among the elderly residents of a local Rhode Island nursing home. Researchers hypothesized that Oscar—the "furry grim reaper" or "four-legged angel of death," as the media called him—could smell ketones, the chemicals released by dying cells, and this drew the feline forecaster toward people in their final days. "I think all animals are able to do things like this," says Dosa, who wrote a book about Oscar that was published last year. But it's not just death that animals can sniff out. A growing number of scientists are now turning to Mother Nature to help with the early detection of a number of ailments, including cancer and tuberculosis. Below are four of the most skilled disease hunters from the animal kingdom. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Drug scandal exposes French regulators to public ire
- Nat Med 17(2):144 (2011)
Nature Medicine | News Drug scandal exposes French regulators to public ire * Barbara Casassus Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:144Year published:(2011)DOI:doi:10.1038/nm0211-144aPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. PARIS — The scandal over the diabetes drug benfluorex, which is alleged to have caused an estimated 500 to 2,000 deaths from valvular heart disease in France, is on course to trigger a major overhaul of the country's drug and health safety system. A devastating 244-page report on the affair, published last month by the governmental General Inspectorate of Social Affairs (IGAS), accused the pharmaceutical company Servier, headquartered in the suburbs of Paris, of misleading authorities about the true nature of benfluorex, which was sold under the brand name Mediator. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - New initiative aims to grow UK life sciences
- Nat Med 17(2):144 (2011)
Nature Medicine | News New initiative aims to grow UK life sciences * Jon Evans Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:144Year published:(2011)DOI:doi:10.1038/nm0211-144bPublished online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Four of the UK's leading healthcare industry associations, encompassing the pharmaceutical, biotechnology, medical device and diagnostic industries, are joining forces under a new initiative called LifeSciencesUK. Launched at the beginning of January, LifeSciencesUK aims to help strengthen and grow the UK life science sector for the benefit of both patients and the UK economy as a whole. The four founding members are the Association of British Healthcare Industries, the Association of the British Pharmaceutical Industry, the BioIndustry Association and the British In Vitro Diagnostics Association. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Genzyme, though unique, could be a bellwether for US biotech
- Nat Med 17(2):145 (2011)
Nature Medicine | News Genzyme, though unique, could be a bellwether for US biotech * Elie Dolgin Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:145Year published:(2011)DOI:doi:10.1038/nm0211-145Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Is Genzyme, a giant among biotechs, lacking a certain je ne sais quoi? Since its launch in 1981, the company has grown from a small, 20-person startup to the biggest biotech corporation in the Boston area, with revenues topping $4 billion annually. But the company has suffered recent setbacks, and last month, as the French drug maker Sanofi-Aventis moved in to take over Genzyme, some analysts saw the possible merger as a sign of how aggressively European pharmaceutical firms are pursuing US biotech companies. Creative Commons View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data * Journal home * Current issue * For authors * Subscribe * E-alert sign up * RSS feed Science jobs from naturejobs * PhD Students in Molecular Medicine * Nordic EMBL Partnership for Molecular Medicine * Helsinki, Finland * Postdoctoral Fellow * Karolinska Institutet * Stockholm, Sweden * PhD Scholarships in Molecular Medicine and Neuroscience * Medical University of Graz * Graz, Austria * Post a free job * More science jobs Open innovation challenges * Delayed Release Formulation for Aqueous Protein Solution Deadline:Apr 02 2011Reward:$40,000 USD A delayed release technology is required for a boosting agent that would allow the simultaneous deli… * Compounds to Combat Citrus Greening Disease Deadline:May 01 2011Reward:$100,000 USD The Seeker, the non-profit Citrus Research and Development Foundation, desires proposals for compoun… * Powered by: * More challenges Top content Emailed * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Detrimental effects of adenosine signaling in sickle cell disease Nature Medicine 19 Dec 2010 * The regulatory subunits of PI3K, p85α and p85β, interact with XBP-1 and increase its nuclear translocation Nature Medicine 28 Mar 2010 * Defying malaria: Arming T cells to halt malaria Nature Medicine 07 Jan 2011 * Putting sleeping sickness to bed Nature Medicine 07 Jan 2011 View all Downloaded * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Personalized investigation Nature Medicine 01 Sep 2010 * The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 Nature Medicine 16 Jan 2011 * The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance Nature Medicine 09 Jan 2011 * Brown adipose tissue activity controls triglyceride clearance Nature Medicine 23 Jan 2011 View all Blogged * A CD8+ T cell transcription signature predicts prognosis in autoimmune disease Nature Medicine 18 Apr 2010 * Is cancer a disease of self-seeding? Nature Medicine 01 Aug 2006 * Limitations of next-generation genome sequence assembly Nature Medicine 21 Nov 2010 * Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes Nature Medicine 05 Dec 2010 * Rapid blue-light–mediated induction of protein interactions in living cells Nature Medicine 31 Oct 2010 View all * Nature Medicine * ISSN: 1078-8956 * EISSN: 1546-170X * About NPG * Contact NPG * RSS web feeds * Help * Privacy policy * Legal notice * Accessibility statement * Terms * Nature News * Naturejobs * Nature Asia * Nature EducationSearch:Go © 2011 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.partner of AGORA, HINARI, OARE, INASP, CrossRef and COUNTER - Vaccines move forward against a range of addictions
- Nat Med 17(2):146 (2011)
Nature Medicine | News Vaccines move forward against a range of addictions * Kelly Rae Chi Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:146Year published:(2011)DOI:doi:10.1038/nm0211-146Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Illicit drug use has reached a near-decade high in the US, so it's perhaps not surprising that public health officials harbor strong hopes for vaccines to treat various addictions. But whereas some experimental vaccines of this sort have shown promise in animal studies and early clinical trials, it seems that some of these shots against addiction—such as immunizations against nicotine—are better positioned to reach the market than others. istockphoto View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data * Journal home * Current issue * For authors * Subscribe * E-alert sign up * RSS feed Science jobs from naturejobs * Stem Cell Biology Scientist * Stanford University * Stanford, CA * PhD Scholarships in Molecular Medicine and Neuroscience * Medical University of Graz * Graz, Austria * PhD Students in Molecular Medicine * Nordic EMBL Partnership for Molecular Medicine * Helsinki, Finland * Post a free job * More science jobs Open innovation challenges * Delayed Release Formulation for Aqueous Protein Solution Deadline:Apr 02 2011Reward:$40,000 USD A delayed release technology is required for a boosting agent that would allow the simultaneous deli… * Low-Volume Liquid Dispersion Mechanism Deadline:Feb 20 2011Reward:$25,000 USD The Seeker is looking for mechanisms to enable dispersion of a fluid sample of 40 nanoliters (nl) ov… * Powered by: * More challenges Top content Emailed * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Detrimental effects of adenosine signaling in sickle cell disease Nature Medicine 19 Dec 2010 * The regulatory subunits of PI3K, p85α and p85β, interact with XBP-1 and increase its nuclear translocation Nature Medicine 28 Mar 2010 * Defying malaria: Arming T cells to halt malaria Nature Medicine 07 Jan 2011 * Putting sleeping sickness to bed Nature Medicine 07 Jan 2011 View all Downloaded * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Personalized investigation Nature Medicine 01 Sep 2010 * The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 Nature Medicine 16 Jan 2011 * The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance Nature Medicine 09 Jan 2011 * Brown adipose tissue activity controls triglyceride clearance Nature Medicine 23 Jan 2011 View all Blogged * A CD8+ T cell transcription signature predicts prognosis in autoimmune disease Nature Medicine 18 Apr 2010 * Is cancer a disease of self-seeding? Nature Medicine 01 Aug 2006 * Limitations of next-generation genome sequence assembly Nature Medicine 21 Nov 2010 * Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes Nature Medicine 05 Dec 2010 * Rapid blue-light–mediated induction of protein interactions in living cells Nature Medicine 31 Oct 2010 View all * Nature Medicine * ISSN: 1078-8956 * EISSN: 1546-170X * About NPG * Contact NPG * RSS web feeds * Help * Privacy policy * Legal notice * Accessibility statement * Terms * Nature News * Naturejobs * Nature Asia * Nature EducationSearch:Go © 2011 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.partner of AGORA, HINARI, OARE, INASP, CrossRef and COUNTER - Straight talk with...Jeremy Berg
- Nat Med 17(2):147 (2011)
Nature Medicine | News Straight talk with...Jeremy Berg * Michelle Pflumm Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:147Year published:(2011)DOI:doi:10.1038/nm0211-147Published online04 February 2011 Abstract Article tools * Full text * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg In December, Jeremy Berg announced plans to step down from the helm of the US National Institute of General and Medical Sciences (NIGMS), the $2 billion branch of the National Institutes of Health charged with funding basic research related to diverse biological processes and diseases. In his seven years as NIGMS director, Berg spearheaded the institute's first formal strategic plan, led efforts to increase workforce diversity and kick-started an open conversation with the scientific community about funding and peer review. In recognition of his many accomplishments, last year Berg was elected to the US Institute of Medicine and awarded a prestigious public service prize from the American Society for Biochemistry and Molecular Biology. At the end of June, Berg will start up a lab studying protein structures at the University of Pittsburgh, as well as serve as the first associate senior vice chancellor for science strategy and planning for Pitt's Schools of the Health Science! s. Ahead of his move, spoke to Berg about his contributions at NIGMS and his plans moving forward. View full text Additional data - News in brief
- Nat Med 17(2):148-149 (2011)
Nature Medicine | News News in brief Journal name:Nature MedicineVolume: 17,Pages:148–149Year published:(2011)DOI:doi:10.1038/nm0211-148Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Policy Playback An analysis of almost 400 worldwide between 1996 and 2009 found that more than half of them occurred in Africa, and about one month elapsed, on average, before the outbreaks there were detected (Proc. Natl. Acad. Sci. USA, 21701–21706, 2010). View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Fecal matters
- Nat Med 17(2):150-152 (2011)
Nature Medicine | News Fecal matters * Roxanne Palmer1 Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Pages:150–152Year published:(2011)DOI:doi:10.1038/nm0211-150Published online04 February 2011 Abstract Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg A dangerous gut disorder is on the rise, and a handful of doctors say that we might be flushing the most effective treatment down the toilet. The procedure they advocate involves transplanting a healthy person's feces into a sick person's colon. Although the approach has shown some promising results, critics worry about unintended consequences—not to mention the 'yuck' factor for patients. reports. View full text Additional data Affiliations * Roxanne Palmer was a news intern with Nature Medicine. She now writes for Law360 in New York. - Review of classification rules represents an opportunity, even for medicine
- Nat Med 17(2):153 (2011)
Nature Medicine | News Review of classification rules represents an opportunity, even for medicine * Steven Aftergood1 Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:153Year published:(2011)DOI:doi:10.1038/nm0211-153Published online04 February 2011 The US system of classifying national security information is bloated and dysfunctional, shielding the government from the scrutiny it needs to avoid error and abusive practices. In the medical field, this secrecy has, in rare cases, enabled unethical, state-sponsored research to proceed without public knowledge. The Fundamental Classification Guidance Review that is now underway could make a difference—if it's done right. View full text Additional data Affiliations * Steven Aftergood directs the Project on Government Secrecy at the Federation of American Scientists and writes the Secrecy News blog. - Corrections
- Nat Med 17(2):149 (2011)
Nature Medicine | Correction Corrections Journal name:Nature MedicineVolume: 17,Page:149Year published:(2011)DOI:doi:10.1038/nm0211-149Published online04 February 2011 Article tools * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg In the January 2011 issue of Nature Medicine, the article entitled "Funds go toward biomedical business incubators in Mexico" (Nat. Med., 7, 2011) specified that UNAM scientists had donated 15 million pesos of their own money into a fund for startup companies; however, the correct amount was 5.5 million pesos. Additionally, the piece incorrectly stated that Biohominis created the H1N1 diagnostic kits, when in fact this was done by scientists at UNAM's Biotechnology Institute. Last, the National Science and Technology Council did not provide funding for the development of the H1N1 kits as stated in the text. The errors have been corrected in the HTML and PDF versions of the article. In the January 2011 issue of Nature Medicine, the article entitled "Scarred by disease" (Nat. Med.17, 18–20, 2011) stated that the STX-100 compound from Stromedix is a small-molecule drug. However, the drug is actually a monoclonal antibody. The error has been corrected in the HTML and PDF versions of the article. Additional data - The tweenage human genome sequence
- Nat Med 17(2):155 (2011)
Nature Medicine | Book Review The tweenage human genome sequence * Ewen Kirkness1 Contact Ewen Kirkness Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:155Year published:(2011)DOI:doi:10.1038/nm0211-155Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Drawing the Map of Life: Inside the Human Genome Project Victor K. McElheny Basic Books, 2010 384 pp., hardcover, $28.00 ISBN: 0465043330 View full text Author information Affiliations * Ewen Kirkness is at The J. Craig Venter Institute, Rockville, Maryland, USA. ekirknes@jcvi.org Competing financial interests The author declares no competing financial interests. Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data * Journal home * Current issue * For authors * Subscribe * E-alert sign up * RSS feed Science jobs from naturejobs * Postdoctoral Fellow * Karolinska Institutet * Stockholm, Sweden * PhD Students in Molecular Medicine * Nordic EMBL Partnership for Molecular Medicine * Helsinki, Finland * Stem Cell Biology Scientist * Stanford University * Stanford, CA * Post a free job * More science jobs Open innovation challenges * Low-Volume Liquid Dispersion Mechanism Deadline:Feb 20 2011Reward:$25,000 USD The Seeker is looking for mechanisms to enable dispersion of a fluid sample of 40 nanoliters (nl) ov… * Derivation of Trophoblast Stem Cells from Human iPS Cells or Human ES Cells Deadline:Mar 13 2011Reward:$50,000 USD The Seeker wishes to derive trophoblast stem (TS) cells from human induced pluripotent stem (iPS) ce… * Powered by: * More challenges Top content Emailed * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Detrimental effects of adenosine signaling in sickle cell disease Nature Medicine 19 Dec 2010 * The regulatory subunits of PI3K, p85α and p85β, interact with XBP-1 and increase its nuclear translocation Nature Medicine 28 Mar 2010 * Defying malaria: Arming T cells to halt malaria Nature Medicine 07 Jan 2011 * Putting sleeping sickness to bed Nature Medicine 07 Jan 2011 View all Downloaded * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Personalized investigation Nature Medicine 01 Sep 2010 * The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 Nature Medicine 16 Jan 2011 * The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance Nature Medicine 09 Jan 2011 * Brown adipose tissue activity controls triglyceride clearance Nature Medicine 23 Jan 2011 View all Blogged * A CD8+ T cell transcription signature predicts prognosis in autoimmune disease Nature Medicine 18 Apr 2010 * Is cancer a disease of self-seeding? Nature Medicine 01 Aug 2006 * Limitations of next-generation genome sequence assembly Nature Medicine 21 Nov 2010 * Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes Nature Medicine 05 Dec 2010 * Rapid blue-light–mediated induction of protein interactions in living cells Nature Medicine 31 Oct 2010 View all - Globular warming: how fat gets to the furnace
- Nat Med 17(2):157-159 (2011)
Nature Medicine | Letter Brown adipose tissue activity controls triglyceride clearance * Alexander Bartelt1 Contact Alexander Bartelt Search for this author in: * NPG journals * PubMed * Google Scholar * Oliver T Bruns2 Search for this author in: * NPG journals * PubMed * Google Scholar * Rudolph Reimer2 Search for this author in: * NPG journals * PubMed * Google Scholar * Heinz Hohenberg2 Search for this author in: * NPG journals * PubMed * Google Scholar * Harald Ittrich3 Search for this author in: * NPG journals * PubMed * Google Scholar * Kersten Peldschus3 Search for this author in: * NPG journals * PubMed * Google Scholar * Michael G Kaul3 Search for this author in: * NPG journals * PubMed * Google Scholar * Ulrich I Tromsdorf4 Search for this author in: * NPG journals * PubMed * Google Scholar * Horst Weller4 Search for this author in: * NPG journals * PubMed * Google Scholar * Christian Waurisch5 Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander Eychmüller5 Search for this author in: * NPG journals * PubMed * Google Scholar * Philip L S M Gordts6 Search for this author in: * NPG journals * PubMed * Google Scholar * Franz Rinninger7 Search for this author in: * NPG journals * PubMed * Google Scholar * Karoline Bruegelmann1 Search for this author in: * NPG journals * PubMed * Google Scholar * Barbara Freund1 Search for this author in: * NPG journals * PubMed * Google Scholar * Peter Nielsen1 Search for this author in: * NPG journals * PubMed * Google Scholar * Martin Merkel1, 8 Search for this author in: * NPG journals * PubMed * Google Scholar * Joerg Heeren1 Contact Joerg Heeren Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorsJournal name:Nature MedicineVolume: 17,Pages:200–205Year published:(2011)DOI:doi:10.1038/nm.2297Received12 October 2010Accepted22 December 2010Published online23 January 2011 Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Brown adipose tissue (BAT) burns fatty acids for heat production to defend the body against cold1, 2 and has recently been shown to be present in humans3, 4, 5. Triglyceride-rich lipoproteins (TRLs) transport lipids in the bloodstream, where the fatty acid moieties are liberated by the action of lipoprotein lipase (LPL)6. Peripheral organs such as muscle and adipose tissue take up the fatty acids, whereas the remaining cholesterol-rich remnant particles are cleared by the liver6. Elevated plasma triglyceride concentrations and prolonged circulation of cholesterol-rich remnants, especially in diabetic dyslipidemia, are risk factors for cardiovascular disease7, 8, 9, 10, 11. However, the precise biological role of BAT for TRL clearance remains unclear. Here we show that increased BAT activity induced by short-term cold exposure controls TRL metabolism in mice. Cold exposure drastically accelerated plasma clearance of triglycerides as a result of increased uptake into BAT, a pr! ocess crucially dependent on local LPL activity and transmembrane receptor CD36. In pathophysiological settings, cold exposure corrected hyperlipidemia and improved deleterious effects of insulin resistance. In conclusion, BAT activity controls vascular lipoprotein homeostasis by inducing a metabolic program that boosts TRL turnover and channels lipids into BAT. Activation of BAT might be a therapeutic approach to reduce elevated triglyceride concentrations and combat obesity in humans. View full text Figures at a glance * Figure 1: Cold exposure modulates fasting and postprandial triglyceride-rich lipoprotein levels. (,) Triglyceride () and cholesterol () FPLC profiles of pooled plasma from fasted FVB mice after 4 h and 24 h cold exposure at 4 °C. () Plasma triglycerides during an oral fat tolerance test in control and cold-exposed FVB mice. () FPLC lipoprotein profiling in control and cold-exposed FVB mice 2 h after an oral fat load. () Organ distribution of triolein-derived 3H-radioactivity in control and cold-exposed FVB mice 2 h after gavage. EpiWAT, epididymal white adipose tissue; SubWAT, subcutaneous white adipose tissue. Mean values ± s.e.m. with n = 12 in and n = 4 in . *P < 0.05; $P < 0.001. * Figure 2: Activated BAT is a central target organ for TRL uptake. (,) Plasma clearance of 59Fe-SPIO () and 3H-triolein-labeled TRLs () in control and cold-exposed C57BL/6 mice. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H-radioactivity () 15 min after intravenous injection. Mean values ± s.e.m. with n ≥ 5. () Representative transversal MRIs of a control and a cold-exposed wild-type FVB mouse before and approximately 10 min after injection of SPIO-labeled TRLs. Arrows in the top images point to BAT, whereas arrows in the bottom images indicate the liver. Scale bar, 1 cm. (,) Coronal MRIs of a representative cold mouse before and 10 min after () and 1 week after () injection of SPIO-TRLs with identical MRI settings. () Representative intravital confocal microscopy images of dissected BAT in a live cold-exposed FVB mouse 2 min (left) and 30 min (right) after QD-TRL (green) injection (arrows indicate QD-TRLs). FITC-dextran (red) to stain blood vessels and DAPI (blue) to label nuclei. Scale bar, 25 μm. () Representative tr! ansmission electron microscopy pictures of high-pressure frozen BAT samples from a SPIO-TRL–injected, cold-exposed FVB mouse. L, lipid droplet; M, mitochondrium; C, capillary. Top left scale bar, 5 μm; top middle scale bar, 1 μm; top right scale bar, 0.05 μm; bottom left scale bar, 1 μm; bottom middle scale bar, 0.05 μm; bottom right scale bar, 0.02 μm. *P < 0.05; &P < 0.01; $P < 0.001. * Figure 3: LPL and CD36 drive TRL clearance into BAT. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H () radioactivity 15 min after intravenous injection in cold FVB mice that were preinjected with tetrahydrolipstatin (THL) to inhibit LPL activity or with heparin to release LPL into circulation, respectively. Mean values ± s.e.m. with n ≥ 5. () Oral fat tolerance test in cold FVB mice pretreated with THL. Mean values ± s.e.m. with n = 5. () Relative mRNA expression in C57BL/6J BAT of several genes. The values for the lean control samples were arbitrarily set to 1 for each gene examined. AU, arbitrary units. () Determination of Cd36 and other fatty acid transporters mRNA copy numbers normalized to the housekeeping TATA-binding protein (Tbp) mRNA by TaqMan. () Consecutive FPLC analysis of TRL-3H-triolein and albumin-3H-oleate in cold-exposed wild-type and Cd36−/− littermates. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H () radioactivity 15 min after intravenous injection of radiolabeled TR! Ls into Cd36−/− and wild-type littermates. Mean values ± s.e.m. with n ≥ 6. *P < 0.05; &P < 0.01; $P < 0.001. * Figure 4: BAT activation corrects hyperlipidemia and is not impaired in insulin resistance. (,) Triglyceride concentrations in hyperlipidemic Apoa5−/− mice during cold exposure () and photograph () of plasma after 24 h cold exposure. () Triglyceride and cholesterol FPLC profiling of pooled plasma from Apoa5−/− mice after 4 h and 24 h cold exposure. () Environmental scanning electron microscopy studies of brown adipose tissue from lean and obese control and cold-exposed mice. Scale bar, 25 μm. () Photographs of interscapular BAT in control and cold-exposed obese mice. Scale bar, 0.5 cm. (,) Combined oral glucose and fat tolerance test in lean and obese control and cold-exposed mice using 14C-deoxyglucose () and 3H-triolein () tracers. (,) Turnover kinetics () and organ uptake () of 3H-triolein-TRLs in lean and obese control and cold-exposed mice. Mean values ± s.e.m. with n = 6. *P < 0.05; &P < 0.01; $P < 0.001. NS, not significant. Author information * Author information * Supplementary information Affiliations * Institute of Biochemistry and Molecular Biology II: Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Alexander Bartelt, * Karoline Bruegelmann, * Barbara Freund, * Peter Nielsen, * Martin Merkel & * Joerg Heeren * Department of Electron Microscopy and Micro Technology, Heinrich-Pette Institute, Hamburg, Germany. * Oliver T Bruns, * Rudolph Reimer & * Heinz Hohenberg * Department of Diagnostic and Interventional Radiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Harald Ittrich, * Kersten Peldschus & * Michael G Kaul * Institute of Physical Chemistry, University of Hamburg, Hamburg, Germany. * Ulrich I Tromsdorf & * Horst Weller * Physical Chemistry, Technical University Dresden, Dresden, Germany. * Christian Waurisch & * Alexander Eychmüller * Department of Human Genetics, University of Leuven, Leuven, Belgium. * Philip L S M Gordts * III. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Franz Rinninger * Asklepios Clinic St. Georg, Department of Internal Medicine, Hamburg, Germany. * Martin Merkel Contributions A.B. and J.H. designed the study, were involved in all aspects of the experiments and co-wrote the manuscript. O.T.B., R.R. and H.H. were responsible for electron microscopy and intravital imaging. H.I., K.P., O.T.B. and M.G.K. were responsible for MRI measurements. C.W., A.E., U.I.T., H.W., B.F. and P.N. were responsible for design and preparation of hydrophobic QD and SPIO, respectively. O.T.B., P.L.S.M.G., F.R., K.B., B.F., P.N. and M.M. were involved in turnover studies. All authors discussed the results and commented on the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding authors Correspondence to: * Alexander Bartelt or * Joerg Heeren Supplementary information * Author information * Supplementary information Movies * Supplementary Video 1 (23M) MRI of SPIO-TRL uptake into BAT.A representative MRI movie of a cold-exposed (cold) and a control mouse (control). After tail vein injection TRL labeled with super-paramagnetic iron-oxide nanocrystals (SPIO-TRL; start of the clock), liver contrast increases as a result of SPIO-TRL uptake in both mice. BAT contrast increase is only observed in the cold mouse. * Supplementary Video 2 (31M) Intravital imaging of BAT after QD-TRL injection. High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey). In cold-exposed mice, TRL which were labeled with hydrophobic fluorescent nanocrystals (QD-TRL; green) are injected via the tail vein. BAT-mediated processing of QD-TRL reveals a rapid attachment to the endothelium. Nuclei are stained with Hoechst (blue) and blood flow is visualized with FITC-dextran (red). * Supplementary Video 3 (45M) Intravital imaging of BODIPY-TRL uptake into BAT. High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey; movie is fourfold accelerated). In cold-exposed mice, TRL which were labeled with BODIPY-TG (BODIPY-TRL, red) are injected via the tail vein at the beginning of the movie. After approx. 2 min, 50 U heparin are injected and initially bound TRL are released from the vessel wall. * Supplementary Video 4 (30M) Intravital imaging of BODIPY-QD-double-labeled TRL uptake into BAT (heparin intervention). High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey). In cold-exposed mice, TRL which were double-labeled with BODIPY-TG and QD (BODIPY-TRL, red; QD-TRL, green) were injected via the tail vein 30 min before the movie starts. At the beginning of the movie only the QD signal is detectable. Next, 50 U heparin are injected, however, the QD signal cannot be released indicating internalization of TRL cores. Thereafter, a second bolus of double-labeled TRL is injected but the particles cannot bind to the endothelium and display prolonged circulation. * Supplementary Video 5 (20M) Intravital imaging of BODIPY-labeled TRL uptake into BAT (heparin intervention). This movie is identical to Supplementary Movie 4 except that only the BODIPY channel (BODIPY-TRL, red) is shown to demonstrate prolonged circulation of TRL while the binding to BAT endothelium is abolished in heparin-treated mice. * Supplementary Video 6 (15M) CD36-deficient mice after cold-exposure #1. A representative movie of a wild-type and Cd36-/- mouse. After 12 h cold exposure, Cd36-/- mice are characterized by low locomotor activity and noticeable shivering compared to wild-type control. * Supplementary Video 7 (26M) CD36-deficient mice after cold-exposure #2. Another example of a wild-type and Cd36-/- mouse. After 12 h cold exposure, Cd36-/- mice are characterized by low locomotor activity and noticeable shivering compared to wild-type control. * Supplementary Video 8 (15M) CD36-deficient mice after recovery. A representative movie of a wild-type and Cd36-/- mouse after 12 h recovery at room temperature. Under these conditions, Cd36-/- mice are indistinguishable from wild-type mice. PDF files * Supplementary Text and Figures (3M) Supplementary Figures and Tables Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Tuberculosis vaccines—a new kid on the block
- Nat Med 17(2):159-160 (2011)
Nature Medicine | Article A multistage tuberculosis vaccine that confers efficient protection before and after exposure * Claus Aagaard1, 6 Contact Claus Aagaard Search for this author in: * NPG journals * PubMed * Google Scholar * Truc Hoang1, 6 Search for this author in: * NPG journals * PubMed * Google Scholar * Jes Dietrich1 Search for this author in: * NPG journals * PubMed * Google Scholar * Pere-Joan Cardona2 Search for this author in: * NPG journals * PubMed * Google Scholar * Angelo Izzo3 Search for this author in: * NPG journals * PubMed * Google Scholar * Gregory Dolganov4 Search for this author in: * NPG journals * PubMed * Google Scholar * Gary K Schoolnik4 Search for this author in: * NPG journals * PubMed * Google Scholar * Joseph P Cassidy5 Search for this author in: * NPG journals * PubMed * Google Scholar * Rolf Billeskov1 Search for this author in: * NPG journals * PubMed * Google Scholar * Peter Andersen1 Contact Peter Andersen Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorsJournal name:Nature MedicineVolume: 17,Pages:189–194Year published:(2011)DOI:doi:10.1038/nm.2285Received12 October 2010Accepted07 December 2010Published online23 January 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg All tuberculosis vaccines currently in clinical trials are designed as prophylactic vaccines based on early expressed antigens. We have developed a multistage vaccination strategy in which the early antigens Ag85B and 6-kDa early secretory antigenic target (ESAT-6) are combined with the latency-associated protein Rv2660c (H56 vaccine). In CB6F1 mice we show that Rv2660c is stably expressed in late stages of infection despite an overall reduced transcription. The H56 vaccine promotes a T cell response against all protein components that is characterized by a high proportion of polyfunctional CD4+ T cells. In three different pre‐exposure mouse models, H56 confers protective immunity characterized by a more efficient containment of late-stage infection than the Ag85B-ESAT6 vaccine (H1) and BCG. In two mouse models of latent tuberculosis, we show that H56 vaccination after exposure is able to control reactivation and significantly lower the bacterial load compared to adjuvant ! control mice. View full text Figures at a glance * Figure 1: Immunogenicity and protective efficacy of H56 and its components. () Purified Ag85B, ESAT-6, Rv2660c and H56 were visualized in Coomassie-stained gels. Ag85B, lane 1; ESAT-6, lane 2; Rv2660c, lane 3 and H56, lane 4. MW, molecular weight. () Protective efficacies in H56/CAF01 vaccinated CB6F1 mice challenged with Mtb (Erdman). One BCG vaccinated group is included as positive control. The differences between vaccinated and nonvaccinated mice 6 weeks after challenge are shown from one of two experiments. () Antigen-specific IFN-γ released from splenocytes purified from Ag85B-, ESAT-6–, Rv2660c- or H56-immunized CB6F1 mice (n = 3 per group) after in vitro stimulation with Ag85B, ESAT-6 or Rv2660c. Results are shown from one of three experiments. () Bacterial numbers in the lung of individual mice (CB6F1) 6 weeks after challenge (n = 6 per group). Data from one of three experiments are shown. () The development of the infection in mice (C57BL/6) immunized with BCG, H56 or saline and challenged with Mtb (H37Rv) was followed by enumerating the! bacilli in the lung of individual mice (n = 6) over a period of 24 weeks. In and , one-way analysis of variance (ANOVA) was used for group comparisons. *P < 0.05; **P < 0.01; ***P < 0.001 compared to the adjuvant control group or BCG (}). Data are means ± s.e.m. * Figure 2: Immune responses and vaccine efficacy of H56 compared to H1. () Antigen-specific IFN-γ release from in vitro–stimulated splenocytes isolated from mice (CB6F1) vaccinated with H1 or H56. Representative results from one of three experiments are shown. *P < 0.05, one-way ANOVA. () IFN-γ released from splenocytes isolated after challenge with Mtb (Erdman) from groups of CB6F1 mice vaccinated with CAF01 adjuvant, BCG, H1 or H56 and stimulated in vitro with the indicated antigens. Results from one of two independent experiments are shown. () The frequency of Ag85B-specific CD4+ T cells (CD44high) producing IFN-γ, TNF-α or IL-2 measured in cells isolated from perfused lungs from mice immunized with CAF01, BCG, H1 or H56. The cytokine profile in individual cells was measured by multicolor flow cytometry by gating for lymphocytes and CD4+ T cells. All possible combinations of cytokine expression were tabulated, and, after subtracting the background (non‐stimulated samples), the results for the seven combinations expressing at least one! of the cytokines are shown. Each of the panels – show results from the same experiment. Two independent experiments were performed. () The bacterial load (CFUs) in the lungs of individual mice. The results are pooled values from two experiments, and each time point represents results from 10–12 mice per group. *P < 0.05; **P < 0.01 using one-way ANOVA. Data are means ± s.e.m. * Figure 3: Evaluation of H56 as a BCG booster. (,) Bacterial numbers (CFUs) 6 () and 24 () weeks after challenge with Mtb (Erdman) in the lung (n = 6 per group) of mice (CB6F1) vaccinated with BCG and boosted twice with either H1 or H56. Representative data from one of two experiments are shown as log10 CFU. ***P < 0.001; **P < 0.01; *P < 0.05, one-way ANOVA with Tukey's post test. Data are means ± s.e.m. * Figure 4: Vaccination with H56 after exposure. () The model used for evaluation of the H56 vaccine after exposure. Mice infected with Mtb (Erdman) were treated with antibiotic for 6 weeks (shaded area). After treatment, mice were killed and bacteria were enumerated in the lungs at the indicated time points (n = 6). The arrows indicate the vaccination time points used in , and . Log10 CFUs are given as mean values ± s.e.m. () IFN-γ released from PBMCs isolated 35 weeks after infection with Mtb (Erdman) from mice vaccinated at weeks 10, 13 and 16 and from nonvaccinated mice (n = 16 per group, pooled PBMCs). PBMCs were stimulated in vitro with Ag85B, ESAT-6 or Rv2660c. () At the same time point, cytokine profiles of antigen-specific CD4+ T cells were measured in Ag85B-stimulated splenocytes by flow cytometry as described in Figure 2c. () The protective efficacy of H56 was measured in two different laboratories. Experiments 1–3 were done in CB6F1 mice at Statens Serum Institut and experiments 4–6 were done in C57BL/6 m! ice at Unitat de Tuberculosi Experimental. In experiments 1–3, bacteria were enumerated in the lungs of individual mice 35 weeks (exp. 1 and 3) and 43 weeks (exp. 2) after challenge. In experiments 1 and 2, mice received two vaccinations, and in experiment 3, three. In experiments 4–6, mice received two vaccinations, and the bacterial load was measured 23 weeks after infection. The CFU values are shown as scattered plots with the median indicated (n = 12–16 per group in each experiment). We used the Mann-Whitney U test for comparison among groups. *P < 0.05; **P < 0.01; ***P < 0.001. Data are means ± s.e.m. Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Claus Aagaard & * Truc Hoang Affiliations * Department of Infectious Disease Immunology, Statens Serum Institut, Copenhagen, Denmark. * Claus Aagaard, * Truc Hoang, * Jes Dietrich, * Rolf Billeskov & * Peter Andersen * Unitat de Tuberculosi Experimental, Institut per a la Investigació en Ciències de la Salut Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona, Catalonia, Spain. * Pere-Joan Cardona * Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA. * Angelo Izzo * Department of Microbiology and Immunology, Stanford University School of Medicine, California, USA. * Gregory Dolganov & * Gary K Schoolnik * Veterinary Sciences Centre, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin, Ireland. * Joseph P Cassidy Contributions C.A. conceived of the study, produced H56, conducted pre‐exposure vaccine studies and prepared the manuscript. T.H. developed the CB6F1 latency model and conducted latency studies in this model. J.D. conducted the BCG boost studies. P.-J.C. developed the C57BL/6 latency model and conducted latency studies in this model. A.I. conducted pre‐exposure vaccine studies. G.D. designed and performed gene expression analyses. G.K.S. designed and performed gene expression analyses. J.P.C. performed histological evaluation of lung specimens. R.B. contributed to the latency vaccine studies. P.A. conceived of the study and prepared the manuscript. All authors discussed the results and commented on the manuscript at all stages. Competing financial interests C.A. and P.A. are co-inventors on a patent application to the Danish patent office covering the use of H56 as a vaccine. All rights have been assigned to Statens Serum Institut, a Danish not-for-profit governmental institute. Corresponding authors Correspondence to: * Claus Aagaard or * Peter Andersen Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (315K) Supplementary Figures 1 and 2 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Sugar rush bleeds the brain
- Nat Med 17(2):161-162 (2011)
Nature Medicine | Letter Hyperglycemia-induced cerebral hematoma expansion is mediated by plasma kallikrein * Jia Liu1, 2, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Ben-Bo Gao1, 2, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Allen C Clermont1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Price Blair3 Search for this author in: * NPG journals * PubMed * Google Scholar * Tamie J Chilcote4 Search for this author in: * NPG journals * PubMed * Google Scholar * Sukanto Sinha4 Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Flaumenhaft3 Search for this author in: * NPG journals * PubMed * Google Scholar * Edward P Feener1, 2 Contact Edward P Feener Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:206–210Year published:(2011)DOI:doi:10.1038/nm.2295Received27 September 2010Accepted16 December 2010Published online23 January 2011 Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Hyperglycemia is associated with greater hematoma expansion and poor clinical outcomes after intracerebral hemorrhage. We show that cerebral hematoma expansion triggered by intracerebral infusion of autologous blood is greater in diabetic rats and mice compared to nondiabetic controls and that this augmented expansion is ameliorated by plasma kallikrein (PK) inhibition or deficiency. Intracerebral injection of purified PK augmented hematoma expansion in both diabetic and acutely hyperglycemic rats, whereas injection of bradykinin, plasmin or tissue plasminogen activator did not elicit such a response. This response, which occurs rapidly, was prevented by co-injection of the glycoprotein VI agonist convulxin and was mimicked by glycoprotein VI inhibition or deficiency, implicating an effect of PK on inhibiting platelet aggregation. We show that PK inhibits collagen-induced platelet aggregation by binding collagen, a response enhanced by elevated glucose concentrations. The ef! fect of hyperglycemia on hematoma expansion and PK-mediated inhibition of platelet aggregation could be mimicked by infusing mannitol. These findings suggest that hyperglycemia auguments cerebral hematoma expansion by PK-mediated osmotic-sensitive inhibition of hemostasis. View full text Figures at a glance * Figure 1: Effect of kallikrein-kinin inhibition on blood-induced hematoma expansion in brains of diabetic animals. (,) Representative images of the dorsal surface () and surface hematoma area as a percentage of hemisphere area () 48 h after intracerebral injection of autologous blood into the right hemisphere or PBS injection (sham) into the left hemisphere in nondiabetic (NDM; n = 15) and diabetic (DM; n = 11) rats. () Representative images of the dorsal surface and surface hematoma area as a percentage of hemisphere area in diabetic Akita mice (Ins2Akita, n = 10) and in nondiabetic littermate wild-type (WT) controls (n = 9) after PBS injection into the left hemisphere and autologous blood injection into the right hemisphere. () Surface hematoma area as a percentage of hemisphere area 48 h after intracerebral injection of autologous blood in nondiabetic rats systemically treated with vehicle (Veh, n = 4) or ASP-440 (440, n = 6) and in diabetic rats systemically treated with vehicle (n = 7), ASP-440 (n = 11), Hoe140 (Hoe, n = 8) or [des-Arg10]-Hoe140 (Des-Hoe, n = 5). () Surface hematoma! area as a percentage of hemisphere area 48 h after intracerebral injection of autologous blood mixed with PK-specific antibody into the right hemisphere or with control IgG into the left hemisphere of diabetic rats (n = 6). () Representative coronal slices 2 h after intracerebral injection of blood into the right hemisphere or PBS injection into the left hemisphere of diabetic Klkb1+/+ and Klkb1−/− mice. () Hemoglobin content of hemispheres subjected to autologous blood in Klkb1+/+ (n = 16), Klkb1+/− (n = 10) and Klkb1−/− (n = 23) mice. () Activated partial thromboplastin time (aPTT) in Klkb1+/+, Klkb1+/− and Klkb1−/− mice (n = 6 per group). *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars in ,,: 2 mm. Error bars represent mean ± s.e.m. * Figure 2: Effect of intracerebral injection of PK on hematoma expansion in brains of diabetic or acutely hyperglycemic rats. (,) Representative images of the dorsal surface and a coronal slice () and surface hematoma area as a percentage of hemisphere area () 48 h after intracerebral injection of PK into the right hemisphere or PBS injection (sham) into the left hemisphere in nondiabetic (NDM; n = 5) and diabetic (DM; n = 7) rats. Scale bar, 2 mm. (,) Hemoglobin () and carbonic anhydrase-1 (CA-1) levels () in a 5-mm coronal section of the hemisphere encompassing the injection site (n = 3–7 per group). () Time course of hematoma expansion after intracerebral injection of PK into the right hemisphere or PBS injection into the left hemisphere of diabetic rats (n = 7–8 per group). () Surface hematoma area as a percentage of hemisphere area in diabetic rats treated with insulin immediately before intracerebral PK injection (n = 8–12 per group). () Time course of blood glucose after intraperitoneal injection of saline (n = 11) or glucose (n = 15) in rats. () Hematoma area as a percentage of hemisp! here area 0.5 h after intracerebral PK or PBS injection in rats intraperitoneally injected with saline or glucose (n = 10–13 per group). () Surface hematoma area as a percentage of hemisphere area in nondiabetic and diabetic rats 0.5 h after intracerebral injection of PK or deactivated PK (De-PK) (n = 6–14 per group). *P < 0.05; **P < 0.01; ***P < 0.001. Error bars represent mean ± s.e.m. * Figure 3: Effect of PK on platelet aggregation and the effect of GPVI on cerebral hematoma expansion in animals. () Effect of PK on collagen-stimulated platelet aggregation. Data represent three independent experiments. () Effect of PK on thrombin- or ADP-induced platelet aggregation. Data represent three independent experiments. () Effect of glucose on PK-induced inhibition of collagen-stimulated platelet aggregation. Data represent three independent experiments. () Effects of PK (160 nM), prekallikrein (160 nM) and deactivated PK (De-PK, 160 nM) on collagen-induced platelet aggregation (n = 4 independent experiments). () Hemisphere hemoglobin content and () hematoma area as a percentage of hemisphere area in C57BL/6 wild-type (WT) mice subjected to JAQ1 or control rat IgG injection in the contralateral hemisphere (n = 6), C57BL/6 WT mice subjected to PBS injection (n = 10) and FcR γ-chain–deficient (Fcer1g−/−) mice subjected to PBS injection (n = 5). () Effect of convulxin on PK-induced hematoma expansion in diabetic rats (n = 6–19 rats per group). () Effects of PK (160 nM) ! and glucose (Glu, 25 mM) on CRP- or convulxin-induced platelet aggregation (n = 3–5 independent experiments). Platelet aggregation was measured with a platelet aggregometer in – and with a microplate reader in and . *P < 0.05; **P < 0.01; ***P < 0.001. Error bars represent mean ± s.e.m. * Figure 4: Effect of osmolarity on the binding of PK to collagen, collagen-stimulated platelet aggregation and hematoma expansion. (,) Sensorgram of the interaction of PK with immobilized collagen type I in the absence () or presence () of glucose. Data represent three independent experiments. () Effect of glucose on PK binding to aorta tissue (n = 5 pieces of rat aorta tissue). () Effects of hyperosmotic mannitol (Man) or hyperosmotic NaCl on PK-induced inhibition of collagen-stimulated platelet aggregation (n = 3 independent experiments). Glu, glucose. () Representative images of the dorsal surface and () surface hematoma area as a percentage of hemisphere area 2 h after intracerebral injection of PK or autologous blood into the right hemisphere and PBS injection (sham) into the left hemisphere of mannitol-treated nondiabetic rats (n = 6 per group). *P < 0.05; **P < 0.01. Scale bar, 2 mm. Error bars represent mean ± s.e.m. Author information * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Jia Liu & * Ben-Bo Gao Affiliations * Research Division, Joslin Diabetes Center, Boston, Massachusetts, USA. * Jia Liu, * Ben-Bo Gao, * Allen C Clermont & * Edward P Feener * Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA. * Jia Liu, * Ben-Bo Gao, * Allen C Clermont & * Edward P Feener * Department of Medicine, Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA. * Price Blair & * Robert Flaumenhaft * ActiveSite Pharmaceuticals, Inc., Berkeley, California, USA. * Tamie J Chilcote & * Sukanto Sinha Contributions J.L. initiated, designed and conducted most of the experiments and wrote the manuscript. B.-B.G. contributed to the design and performance of animal studies, biochemical analyses, platelet studies and manuscript writing. A.C.C. contributed to animal studies. P.B. and R.F. designed and performed studies using platelet aggregometer and contributed to data interpretation. T.J.C. and S.S. contributed to data interpretation and manuscript editing. E.P.F. designed and supervised the entire study and contributed to manuscript writing. Competing financial interests Tamie J. Chilcote and Sukanto Sinha are employees of and shareholders in ActiveSite Pharmaceuticals, which provided ASP-440. The Joslin Diabetes Center and ActiveSite Pharmaceuticals have submitted a patent application to the US Patent and Trademark Office and the European Patent Office on the methods for treatment of kallikrein-related disorders. Corresponding author Correspondence to: * Edward P Feener Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (274K) Supplementary Figures 1–8 and Supplementary Methods Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Stemming a tumor with a little miR
- Nat Med 17(2):162-164 (2011)
Nature Medicine | Letter The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 * Can Liu1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Kevin Kelnar3 Search for this author in: * NPG journals * PubMed * Google Scholar * Bigang Liu1 Search for this author in: * NPG journals * PubMed * Google Scholar * Xin Chen1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Tammy Calhoun-Davis1 Search for this author in: * NPG journals * PubMed * Google Scholar * Hangwen Li1 Search for this author in: * NPG journals * PubMed * Google Scholar * Lubna Patrawala2 Search for this author in: * NPG journals * PubMed * Google Scholar * Hong Yan1 Search for this author in: * NPG journals * PubMed * Google Scholar * Collene Jeter1 Search for this author in: * NPG journals * PubMed * Google Scholar * Sofia Honorio1 Search for this author in: * NPG journals * PubMed * Google Scholar * Jason F Wiggins3 Search for this author in: * NPG journals * PubMed * Google Scholar * Andreas G Bader3 Search for this author in: * NPG journals * PubMed * Google Scholar * Randy Fagin4 Search for this author in: * NPG journals * PubMed * Google Scholar * David Brown3 Search for this author in: * NPG journals * PubMed * Google Scholar * Dean G Tang1, 2 Contact Dean G Tang Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:211–215Year published:(2011)DOI:doi:10.1038/nm.2284Received14 May 2010Accepted30 November 2010Published online16 January 2011 Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Cancer stem cells (CSCs), or tumor-initiating cells, are involved in tumor progression and metastasis1. MicroRNAs (miRNAs) regulate both normal stem cells and CSCs2, 3, 4, 5, and dysregulation of miRNAs has been implicated in tumorigenesis6. CSCs in many tumors—including cancers of the breast7, pancreas8, head and neck9, colon10, 11, small intestine12, liver13, stomach14, bladder15 and ovary16—have been identified using the adhesion molecule CD44, either individually or in combination with other marker(s). Prostate CSCs with enhanced clonogenic17 and tumor-initiating and metastatic18, 19 capacities are enriched in the CD44+ cell population, but whether miRNAs regulate CD44+ prostate cancer cells and prostate cancer metastasis remains unclear. Here we show, through expression analysis, that miR-34a, a p53 target20, 21, 22, 23, 24, was underexpressed in CD44+ prostate cancer cells purified from xenograft and primary tumors. Enforced expression of miR-34a in bulk or purifie! d CD44+ prostate cancer cells inhibited clonogenic expansion, tumor regeneration, and metastasis. In contrast, expression of miR-34a antagomirs in CD44− prostate cancer cells promoted tumor development and metastasis. Systemically delivered miR-34a inhibited prostate cancer metastasis and extended survival of tumor-bearing mice. We identified and validated CD44 as a direct and functional target of miR-34a and found that CD44 knockdown phenocopied miR-34a overexpression in inhibiting prostate cancer regeneration and metastasis. Our study shows that miR-34a is a key negative regulator of CD44+ prostate cancer cells and establishes a strong rationale for developing miR-34a as a novel therapeutic agent against prostate CSCs. View full text Figures at a glance * Figure 1: Underexpression and tumor-inhibitory effects of miR-34a. () Experimental scheme. (,) miR-34a levels in CD44+ xenograft (, PC4, LAPC4) or primary tumor (, HPCa) cells (mean percentage of marker-positive over marker-negative cells). (,) Orthotopic LAPC9 () and subcutaneous HPCa58 (; black, lenti-ctl; gray, lenti-34a; n = 10 and n = 7 for primary and secondary experiments, respectively) tumor growth (mean ± s.d.). () Lenti-34a mediated overexpression of miR-34a in purified CD44+ Du145 cells completely blocked subcutaneous tumor regeneration. () Subcutaneous tumor regeneration from purified CD44+ LAPC9 cells transfected with miR-NC or miR-34a. () Anti-miR-34a oligo transfection in purified CD44− Du145 cells promoted subcutaneous tumor growth. () Weight (mean ± s.d.) of LAPC9 tumors from bulk cells transfected with anti-NC or anti-34a oligos and implanted (100,000 cells) in the DP. Mice were killed at day 46. () Representative microphotographs showing increased lung metastasis by anti-34a (top, animal number and tumor weight; scale! bar, 100 μm; also see Supplementary Fig. 7c,d). * Figure 2: miR-34a inhibits prostate CSC properties. () Holoclone assays in Du145 cells. Cells transfected with miR-NC (NC) or miR-34a (34a) oligos were used in three experiments (Exp. I, 100 cells per well scored on day 9; Exp. II, 100 cells per well scored on day 13; Exp. III, 500 cells per well scored on day 7). () Clonogenic assays in Du145 cells. Cells (3,000 per well) were plated in Matrigel and colonies counted on day 13. NT, nontransfected. () Matrigel clonogenic assays in LAPC4 cells. Two experiments were performed (Exp. I, 1,250 cells per well scored on day 5, *P = 0.005; Exp. II, 25,000 cells per well scored on day 5, **P = 0.015). () Sphere assays in LAPC4 cells infected with lenti-ctl (C) or lenti-34a. Both primary and secondary spheres were scored on day 15. () Holoclone assays in PPC-1 cells (quantified on day 5). () Sphere assays in HPCa101 cells infected with lenti-ctl (C) or lenti-34a. Both primary and secondary spheres were scored 3 weeks later. (,) Sphere assays in purified CD44+ HPCa116 cells transfected w! ith NC or miR-34a oligos () or CD44− HPCa116 cells transfected with anti-NC or anti-34a oligos (). Spheres were scored on day 15. * Figure 3: Therapeutic effects of miR-34a. () Injections of miR-34a into the tail vein inhibited orthotopic PC3 tumor growth (n = 9 each). (–) Injections of miR-34a oligos into the tail vein inhibited metastasis (GFP+ foci in the endpoint lungs; mean ± s.d., n = 6 per group) of orthotopic LAPC9-GFP tumors () without significantly affecting tumor growth () and extended mouse survival (; Kaplan-Meier analysis and log-rank test). (,) The fourth set of therapeutic experiments in LAPC9 cells. Representative lung images (, animal number and tumor weight indicated on top; scale bar, 100 μm) and quantification of lung metastases (; mean ± s.d., n = 10 per group). * Figure 4: CD44 is a direct and functional target of miR-34a. () Representative CD44 immunohistochemistry images in Du145 tumors from cells infected with MSCV-PIG (control) or MSCV-34a vectors (western blot on the right) and PC3 tumors harvested from mice treated with miR-NC or miR-34a oligos. Scale bars, 10 μm. () miR-34a downregulates CD44 in Du145 (left) and PPC-1 (right) cells. Relative levels of CD44 indicated at the bottom. () Schematic of two putative miR-34a binding sites in the CD44 3′ UTR. () Luciferase experiments in Du145 cells (*P < 0.01). () Knockdown of CD44 inhibits LAPC4 tumor regeneration (Supplementary Fig. 12). (,) Knockdown of CD44 inhibits PC3 cell metastasis; shown are quantification () and images (; scale bar, 100 μm). (,) Invasion assays. miR-34a oligos inhibited Matrigel invasion of CD44+ Du145 cells (), and this inhibition was partially overcome by overexpression of a human CD44 cDNA lacking the miR-34a binding sites at the 3′ UTR (). Invasion expressed as values relative to the corresponding controls. ! () A schematic summary. The part highlighted in red refers to the novel findings of this study. Author information * Author information * Supplementary information Affiliations * Department of Molecular Carcinogenesis, the University of Texas M.D. Anderson Cancer Center, Science Park, Smithville, Texas, USA. * Can Liu, * Bigang Liu, * Xin Chen, * Tammy Calhoun-Davis, * Hangwen Li, * Hong Yan, * Collene Jeter, * Sofia Honorio & * Dean G Tang * Program in Molecular Carcinogenesis, The University of Texas Graduate School of Biomedical Sciences (GSBS), Houston, Texas, USA. * Can Liu, * Xin Chen, * Lubna Patrawala & * Dean G Tang * Mirna Therapeutics, Inc., Austin, Texas, USA. * Kevin Kelnar, * Jason F Wiggins, * Andreas G Bader & * David Brown * The Hospital at Westlake, Austin, Texas, USA. * Randy Fagin Contributions C.L., K.K., B.L., X.C. and L.P. designed and performed the experiments with help from C.J., T.C.-D., H.L., S.H., H.Y., J.F.W. and A.G.B., R.F. provided all HPCa samples. C.L. and D.G.T. prepared the manuscript. D.G.T., with help from D.B., designed the experiments and supervised the whole project. All authors discussed the results and commented on the manuscript. Competing financial interests K.K, J.F.W, A.G.B. and D.B are employees of Mirna Therapeutics, Inc., which develops miRNA-based therapeutics. Corresponding author Correspondence to: * Dean G Tang Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (2M) Supplementary Results, Supplementary Methods, Supplementary Figures 1–15 and Supplementary Tables 1 and 2 Read the full article * Instant access to this article: US$18Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Linking the inflammasome to obesity-related disease
- Nat Med 17(2):164-165 (2011)
Nature Medicine | Article The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance * Bolormaa Vandanmagsar1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Yun-Hee Youm1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Anthony Ravussin1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Jose E Galgani2, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Krisztian Stadler2, 4 Search for this author in: * NPG journals * PubMed * Google Scholar * Randall L Mynatt2 Search for this author in: * NPG journals * PubMed * Google Scholar * Eric Ravussin2, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Jacqueline M Stephens5 Search for this author in: * NPG journals * PubMed * Google Scholar * Vishwa Deep Dixit1, 2 Contact Vishwa Deep Dixit Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:179–188Year published:(2011)DOI:doi:10.1038/nm.2279Received18 May 2010Accepted18 November 2010Published online09 January 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The emergence of chronic inflammation during obesity in the absence of overt infection or well-defined autoimmune processes is a puzzling phenomenon. The Nod-like receptor (NLR) family of innate immune cell sensors, such as the nucleotide-binding domain, leucine-rich–containing family, pyrin domain–containing-3 (Nlrp3, but also known as Nalp3 or cryopyrin) inflammasome are implicated in recognizing certain nonmicrobial originated 'danger signals' leading to caspase-1 activation and subsequent interleukin-1β (IL-1β) and IL-18 secretion. We show that calorie restriction and exercise-mediated weight loss in obese individuals with type 2 diabetes is associated with a reduction in adipose tissue expression of Nlrp3 as well as with decreased inflammation and improved insulin sensitivity. We further found that the Nlrp3 inflammasome senses lipotoxicity-associated increases in intracellular ceramide to induce caspase-1 cleavage in macrophages and adipose tissue. Ablation of Nl! rp3 in mice prevents obesity-induced inflammasome activation in fat depots and liver as well as enhances insulin signaling. Furthermore, elimination of Nlrp3 in obese mice reduces IL-18 and adipose tissue interferon-γ (IFN-γ) expression, increases naive T cell numbers and reduces effector T cell numbers in adipose tissue. Collectively, these data establish that the Nlrp3 inflammasome senses obesity-associated danger signals and contributes to obesity-induced inflammation and insulin resistance. View full text Figures at a glance * Figure 1: Reduction of Nlrp3 and IL-1β expression is associated with improvement of insulin sensitivity. (,) Positive correlation of the visceral fat mRNA expression of Il1b () and Nlrp3 () with body weight of C57BL/6 mice (n = 32); Pearson's correlations are r = 0.364, P = 0.0178 for Il1b and r = 0.672, P < 0.0001 for Nlrp3, respectively. (–) Il1b (), Nlrp3 () and Pycard () mRNA in visceral and subcutaneous adipose tissue from ad libitum chow-fed control (AL) and 40% calorie-restricted (CR) 12-month-old mice, n = 6; *P < 0.01, **P < 0.005. (,) Representative H&E staining showing adipocyte size in visceral () and subcutaneous () fat tissue from ad libitum–fed control and calorie-restricted 12-month-old C57BL/6 mice. () IL1B (left), NLRP3 (middle) and PYCARD (right) gene expression, as examined by quantitative RT-PCR (qRT-PCR) in human SAT in obese individuals with T2DM before and after 1-year weight loss. *P = 0.01, **P = 0.001. () Positive correlation of changes in gene expression of IL1B and NLRP3 in human abdominal subcutaneous fat with changes in fasting glucose level f! rom baseline to 1 year after intervention; Pearson's correlations are r = 0.53, P = 0.12 for IL1B and r = 0.69, P = 0.03 for NLRP3. Relative gene expression levels are depicted as means ± s.e.m. n = 10. * Figure 2: Elimination of Nlrp3 expression prevents obesity-induced caspase-1 cleavage and IL-1β and IL-18 activation. () Immunofluorescence staining of epididymal fat (eFat) tissue sections stained with antibodies against F4/80 (red) and Nlrp3 (green). Merge of images with nuclear staining with DAPI shows colocalization of Nlrp3 with ATM (yellow arrowheads). Negative control staining with antibody to Nlrp3 together with antibody to F4/80 in adipose tissue of Nlrp3−/− mice shows reduced Nlrp3-specific immunostaining. (,) The qRT-PCR analysis of Nlrp3 () and Pycard () mRNA in purified ATM and SVF cells derived from SAT and VAT of 6-month-old DIO mice, and mature 3T3-L1 adipocytes (Adip). () Immunoblot analysis showing the kinetics of caspase-1 (p20) cleavage and active IL-1β (p17) accumulation in adipose tissue of mice at various stages of diet-induced obesity. The age, in months, of the mice at the time of tissue harvesting is indicated. () Western blot analysis of activated caspase-1 (p20) in VAT, SAT and liver tissues from 9-month-old DIO-WT (C57BL/6) and DIO-Nlrp3−/− mice. Result! s shown are representative of three independent experiments. MW, molecular weight. () Western blot analysis of activated caspase-1 (p20) in kidney from 9-month-old DIO-WT and DIO-Nlrp3−/− mice. () Western blot analysis of IL-1β activation in adipose tissue of 6- and 7-month-old DIO mice. () Serum IL-18 concentration in age-matched WT and Nlrp3−/− mice fed normal chow or 60% HFD, starting at 2 months of age for all groups, for 4 months and 7 months. All data are presented as means ± s.e.m., n = 6–10 mice; *P < 0.05. * Figure 3: The Nlrp3 inflammasome regulates insulin sensitivity and steatohepatitis in obesity. () Insulin tolerance test (ITT) and glucose tolerance test (GTT) in male WT and Nlrp3−/− DIO mice fed 60% HFD for 6 weeks. (,) The ITT and GTT in 6-month-old () and 9-month-old () WT and Nlrp3−/− DIO mice showing the area under the curve (green for WT and blue for Nlrp3−/−) with trend lines (n = 5–7 per group). () The total area under the curve (AUC) for ITT in 6- (left) and 9-month-old (right) WT and Nlrp3−/− DIO mice. (*P < 0.01). () Adipocyte size in visceral fat tissue from 8-month-old DIO-WT (Nlrp3+/+) and Nlrp3−/− mice (H&E staining). Representative micrographs and the quantified results are shown. n = 6; *P < 0.001. () Akt, IRS-1 and Erk-1/2 signaling in vivo, as determined by western blotting for phosphorylation, in VAT, SAT, liver and muscle from the 8-month-old DIO Nlrp3+/+ and DIO Nlrp3−/− mice. The Ser322/338 antibody detects phosphorylation at Ser332 and Ser336. () H&E staining of sections from liver of 9-month-old DIO WT and Nlrp3−/�! �� mice. () Fatty acid oxidation and fatty acid synthesis gene expression, as examined by qRT-PCR in the liver of 9-month-old DIO WT and Nlrp3−/− mice. mRNA expression was normalized to glyceraldehyde 3-phosphate dehydrogenase (Gapdh) expression and is shown as fold change (ΔΔCt) with the values for WT arbitrarily set to 1. All data are presented as means ± s.e.m. (n = 5), *P < 0.05, **P < 0.001. * Figure 4: Nlrp3 senses ceramide to induce IL-1β and regulates adipose tissue macrophage activation in obesity. () The cell extracts of LPS-primed bone marrow–derived macrophages (BMDMs) from Nlrp3+/+ and Nlrp3−/− mice (n = 6–9) were analyzed for caspase-1 active form by western blotting as indicated. () The BMDM cells were stimulated with LPS and C2 ceramide (0.1 mM), and cell supernatants were analyzed for IL-1β. Results are representative of three separate experiments. () Epididymal adipose tissue explants from 9-month-old Nlrp3+/+ and Nlrp3−/− DIO mice were cultured for 24 h in the presence of LPS, C2 ceramide or both. Caspase-1 activation was determined by immunoblot analysis. () M1- and M2-associated gene expression, as examined by qRT-PCR in macrophages originating from VAT and SAT of 9-month-old DIO-Nlrp3+/+ and DIO-Nlrp3−/− mice. The mRNA expression was normalized to glyceraldehydes 3-phosphate dehydrogenase (Gapdh) and shown as fold change (ΔΔCt) with the values for WT arbitrarily set to 1. All data are presented as means ± s.e.m., *P < 0.01, **P < 0.001. * Figure 5: Ablation of the Nlrp3 inflammasome reduces adipose tissue effector T cells without affecting Treg cells in visceral fat of obese mice. () FACS plot of SVF cells isolated from VAT of 9-month-old WT and Nlrp3−/− obese mice. The dot plots depict FSC and SSC (left) and the sequential gating strategy for analysis of ATMs and T cells. Gate 1 (top) of larger cells shows the presence of F4/80+ cells (histogram) and expression of macrophage markers CD206 and CD11c on ATMs. Gating of smaller cells (gate 2, bottom) reveals the absence of ATMs in this population of SVF. () The gate 2 (lymphoid gate) showing CD4+ and CD8+ T cells in SVF. CD4+ and CD8+ cells were evaluated for naive T cells (CD62L+CD44−, blue boxes) and effector memory (CD62L−CD44+, red boxes) CD4+ and CD8+ T cells. The FACS analysis was repeated in three independent pooled SVF fractions from a total of 12–14 mice and percent gated cell frequencies are indicated in each representative dot plot. () Gated percentage and absolute numbers (in million cells) of naive (CD62L+CD44−) and effector memory (CD62L−CD44+) CD4+ and CD8+ T cells. () Numbe! r of stromal vascular cells per gram of fat tissue (n = 4–6) in 3- and 9-month-old WT and Nlrp3−/− DIO mice. () Representative FACS plots showing CD4+CD25+Foxp3+ T regulatory cells in VAT of 9-month-old WT and Nlrp3−/− DIO mice. () Gated percentage of Treg cells in VAT and SAT of 9-month-old WT and Nlrp3−/− DIO mice (n = 6 per group). All data are presented as means ± s.e.m., *P < 0.05. * Figure 6: Elimination of the Nlrp3 inflammasome reduces obesity-induced macrophage–mediated T cell activation in adipose tissue. () Representative FACS plots of SVF cells from SAT of 6- and 9-month-old WT and Nlrp3−/− DIO mice stained with CD206 and CD11c. () The gated percentage of CD11c−CD206+ M2 cells in SAT of 9-month-old obese mice. () Dot plots showing naive (CD62L+CD44−, blue boxes) and effector memory (CD62L−CD44+, red boxes) CD4+ and CD8+ T cells in SVF from 9-month-old obese WT and Nlrp3−/− mice. The percent gated cell frequencies are indicated in each representative dot plot () Gated percentage and absolute numbers (in million cells) of naive (CD62L+CD44−) and effector memory (CD62L−CD44+) CD4+ and CD8+ T cells. () mRNA level of the TH1 cytokine Ifnγ in VAT and SAT of 9-month-old obese WT and Nlrp3−/− mice as determined by qRT–PCR. () IFN-γ (19-kDa) in VAT of 9-month-old WT and Nlrp3−/− DIO mice, as examined by western blotting. () IP10 and MCP-1 levels in the serum of 9-month-old lean WT and obese WT and Nlrp3−/− mice (n = 5). The data shown are means ± s! .e.m., *P < 0.05. () Hypothetical model of Nlrp3 inflammasome activation in obesity. In the absence of danger signals in the healthy lean state, tissue-resident macrophages and T cells may participate in the maintenance of adipose tissue function. In obesity, the Nlrp3 inflammasome senses the obesity-associated danger signals such as ceramides leading to caspase-1 autoactivation and IL-1β and IL-18 production from ATMs. Secondary signals from activated ATMs to effector adipose T cells (defined as CD44+CD62L−) sustain the reciprocal proinflammatory feed-forward cascade during obesity, leading to insulin resistance. ROS, reactive oxygen species. Author information * Abstract * Author information * Supplementary information Affiliations * Laboratory of Neuroendocrine-Immunology, Louisiana State University, Baton Rouge, Louisiana, USA. * Bolormaa Vandanmagsar, * Yun-Hee Youm, * Anthony Ravussin & * Vishwa Deep Dixit * Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, Louisiana, USA. * Bolormaa Vandanmagsar, * Yun-Hee Youm, * Anthony Ravussin, * Jose E Galgani, * Krisztian Stadler, * Randall L Mynatt, * Eric Ravussin & * Vishwa Deep Dixit * Human Physiology Laboratory, Louisiana State University, Baton Rouge, Louisiana, USA. * Jose E Galgani & * Eric Ravussin * Oxidative Stress and Disease Laboratory, Louisiana State University, Baton Rouge, Louisiana, USA. * Krisztian Stadler * Department of Biological Sciences, Louisiana State University System, Baton Rouge, Louisiana, USA. * Jacqueline M Stephens Contributions B.V. performed real-time PCRs, flow cytometry assays, adipose tissue macrophage selections, some western blots, ITT, GTT and cytokine assays, managed the transgenic animal colony and participated in experimental design, data analysis and manuscript preparation. Y.-H.Y. performed all caspase-1 and IL1β western blots, adipose and liver histologies, and macrophage culture experiments and analyzed the data. A.R. Performed body composition analysis, tissue collections, lipid analysis, animal husbandry, genotyping, ITT, GTT and adipocyte size measurement. J.E.G. and E.R. designed and supervised the human studies, analyzed the glucose and insulin sensitivity data in obese T2DM subjects and discussed the hypotheses. K.S. performed the caspase-1 western blot in the kidneys of HFD fed WT and Nlrp3 null mice. R.L.M. participated in standardizing the ITT and GTT assays and advised on the design of experiments for liver fatty acid synthesis and oxidation gene expression. J.M.S. performe! d the western blots for some of the insulin-signaling experiments, helped with data interpretation, discussed the hypotheses and participated in manuscript preparation. V.D.D. conceived the project, designed the experiments, performed some of the cytokine assays and flow cytometry, helped with data interpretation, participated in data analysis, directed the project and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * Vishwa Deep Dixit Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (376K) Supplementary Figures 1–7 and Supplementary Table 1 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Cystic fibrosis: an-ion transport issue?
- Nat Med 17(2):166-167 (2011)
Nature Medicine | Community Corner Cystic fibrosis: an-ion transport issue? Journal name:Nature MedicineVolume: 17,Pages:166–167Year published:(2011)DOI:doi:10.1038/nm0211-166Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Medi-Mation Ltd/Photo Researchers, Inc. Defective anion transport may be the initiating event in cystic fibrosis lung disease. View full text Author information Article tools * Full text * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Competing financial interests The author declares no competing financial interests. Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data * Journal home * Current issue * For authors * Subscribe * E-alert sign up * RSS feed Science jobs from naturejobs * PhD Scholarships in Molecular Medicine and Neuroscience * Medical University of Graz * Graz, Austria * Postdoctoral Fellow * Karolinska Institutet * Stockholm, Sweden * PhD Students in Molecular Medicine * Nordic EMBL Partnership for Molecular Medicine * Helsinki, Finland * Post a free job * More science jobs Open innovation challenges * Delayed Release Formulation for Aqueous Protein Solution Deadline:Apr 02 2011Reward:$40,000 USD A delayed release technology is required for a boosting agent that would allow the simultaneous deli… * Derivation of Trophoblast Stem Cells from Human iPS Cells or Human ES Cells Deadline:Mar 13 2011Reward:$50,000 USD The Seeker wishes to derive trophoblast stem (TS) cells from human induced pluripotent stem (iPS) ce… * Powered by: * More challenges Top content Emailed * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Detrimental effects of adenosine signaling in sickle cell disease Nature Medicine 19 Dec 2010 * The regulatory subunits of PI3K, p85α and p85β, interact with XBP-1 and increase its nuclear translocation Nature Medicine 28 Mar 2010 * Defying malaria: Arming T cells to halt malaria Nature Medicine 07 Jan 2011 * Putting sleeping sickness to bed Nature Medicine 07 Jan 2011 View all Downloaded * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Personalized investigation Nature Medicine 01 Sep 2010 * The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 Nature Medicine 16 Jan 2011 * The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance Nature Medicine 09 Jan 2011 * Brown adipose tissue activity controls triglyceride clearance Nature Medicine 23 Jan 2011 View all Blogged * A CD8+ T cell transcription signature predicts prognosis in autoimmune disease Nature Medicine 18 Apr 2010 * Is cancer a disease of self-seeding? Nature Medicine 01 Aug 2006 * Limitations of next-generation genome sequence assembly Nature Medicine 21 Nov 2010 * Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes Nature Medicine 05 Dec 2010 * Rapid blue-light–mediated induction of protein interactions in living cells Nature Medicine 31 Oct 2010 View all * Nature Medicine * ISSN: 1078-8956 * EISSN: 1546-170X * About NPG * Contact NPG * RSS web feeds * Help * Privacy policy * Legal notice * Accessibility statement * Terms * Nature News * Naturejobs * Nature Asia * Nature EducationSearch:Go © 2011 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.partner of AGORA, HINARI, OARE, INASP, CrossRef and COUNTER - A MRSA-terious enemy among us: Boosting MRSA vaccines
- Nat Med 17(2):168-169 (2011)
Nature Medicine | Between Bedside and Bench A MRSA-terious enemy among us: Boosting MRSA vaccines * Scott D Kobayashi1 Search for this author in: * NPG journals * PubMed * Google Scholar * Frank R DeLeo1 Contact Frank R DeLeo Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:168–169Year published:(2011)DOI:doi:10.1038/nm0211-168Published online04 February 2011 Infection with methicillin-resistant Staphylococcus aureus (MRSA) can cause symptoms ranging from mild skin infections to more severe disease in various organs, even among healthy individuals. The ability of this pathogen to escape our immune arsenal and overcome antibiotic therapy poses a challenge to preventing their spread and treating the related symptoms. In 'Bench to Bedside', Scott Kobayashi and Frank DeLeo explore new approaches for vaccine development that focus on antigens required for establishment of disease. Studies with infected mice immunized against S. aureus coagulases—important for abscess formation and bloodstream infection—suggest such an approach may be used to reduce bacterial load and protect against severe disease in humans. In 'Bedside to Bench', Michael Otto examines a large human study where the presence of genes encoding Panton-Valentine leukocidin toxin (PVL) in community-associated MRSA did not correlate with complicated skin structure infec! tions—a result opposing the widespread notion that PVL is the primary CA-MRSA virulence factor. View full text Author information Article tools * Full text * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Affiliations * Scott D. Kobayashi and Frank R. DeLeo are at the Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, US National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * Frank R DeLeo Additional data - A MRSA-terious enemy among us: End of the PVL controversy?
- Nat Med 17(2):169-170 (2011)
Nature Medicine | Between Bedside and Bench A MRSA-terious enemy among us: End of the PVL controversy? * Michael Otto1 Contact Michael Otto Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Pages:169–170Year published:(2011)DOI:doi:10.1038/nm0211-169Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Bench to Bedside CA-MRSA strains are causing an ongoing epidemic of skin and soft tissue infections, which characteristically develop in healthy individuals without predisposing conditions. The US has experienced the largest CA-MRSA outbreak, with one strain, USA300, being responsible for the vast majority of infections. In 2005, 13.7% of all invasive MRSA infections in the US were community associated1. Despite the obvious clinical importance of this disease, the molecular basis underlying the success of CA-MRSA strains as pathogens has remained obscure. Frank DeLeo View full text Author information Article tools * Full text * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Affiliations * Michael Otto is at the US National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. Competing financial interests The author declares no competing financial interests. Corresponding author Correspondence to: * Michael Otto Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data * Journal home * Current issue * For authors * Subscribe * E-alert sign up * RSS feed Science jobs from naturejobs * Postdoctoral Fellow * Karolinska Institutet * Stockholm, Sweden * PhD Scholarships in Molecular Medicine and Neuroscience * Medical University of Graz * Graz, Austria * Junior and Senior Scientists * Medical University of South Carolina * Charleston, SC * Post a free job * More science jobs Open innovation challenges * RNAi Sequences Targeted to the Asian Citrus Psyllid Genome Deadline:May 03 2011Reward:$100,000 USD The non-profit Citrus Research and Development Foundation, desires proposals for RNA sequences that … * Derivation of Trophoblast Stem Cells from Human iPS Cells or Human ES Cells Deadline:Mar 13 2011Reward:$50,000 USD The Seeker wishes to derive trophoblast stem (TS) cells from human induced pluripotent stem (iPS) ce… * Powered by: * More challenges Top content Emailed * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Detrimental effects of adenosine signaling in sickle cell disease Nature Medicine 19 Dec 2010 * The regulatory subunits of PI3K, p85α and p85β, interact with XBP-1 and increase its nuclear translocation Nature Medicine 28 Mar 2010 * Defying malaria: Arming T cells to halt malaria Nature Medicine 07 Jan 2011 * Putting sleeping sickness to bed Nature Medicine 07 Jan 2011 View all Downloaded * A multistage tuberculosis vaccine that confers efficient protection before and after exposure Nature Medicine 23 Jan 2011 * Personalized investigation Nature Medicine 01 Sep 2010 * The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 Nature Medicine 16 Jan 2011 * The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance Nature Medicine 09 Jan 2011 * Brown adipose tissue activity controls triglyceride clearance Nature Medicine 23 Jan 2011 View all Blogged * A CD8+ T cell transcription signature predicts prognosis in autoimmune disease Nature Medicine 18 Apr 2010 * Is cancer a disease of self-seeding? Nature Medicine 01 Aug 2006 * Limitations of next-generation genome sequence assembly Nature Medicine 21 Nov 2010 * Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes Nature Medicine 05 Dec 2010 * Rapid blue-light–mediated induction of protein interactions in living cells Nature Medicine 31 Oct 2010 View all * Nature Medicine * ISSN: 1078-8956 * EISSN: 1546-170X * About NPG * Contact NPG * RSS web feeds * Help * Privacy policy * Legal notice * Accessibility statement * Terms * Nature News * Naturejobs * Nature Asia * Nature EducationSearch:Go © 2011 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.partner of AGORA, HINARI, OARE, INASP, CrossRef and COUNTER - Research Highlights
- Nat Med 17(2):172-173 (2011)
Nature Medicine | Research Highlights Research Highlights Journal name:Nature MedicineVolume: 17,Pages:172–173Year published:(2011)DOI:doi:10.1038/nm0211-172Published online04 February 2011 Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Immunology: Layered lymphocytes The human fetal immune system derives from a different source of cells than the adult equivalent, according to a new study (Science, 1695–1699, 2010). The findings, which show that immune cell development is layered as two distinct waves, challenge the conventional wisdom that T lymphocytes form in a linear fashion. Jeff Mold et al. compared adult T cells with those taken from 18- to 22-week-old fetuses and observed consistent differences in the gene expression profiles of the cells. The researchers next implanted hematopoietic stem cells from various human developmental stages into immunocompromised mice and found that fetal stem cells showed a greater propensity for generating regulatory T cells during thymic maturation. View full text Read the full article * Instant access to this article: US$32Buy now * Subscribe to Nature Medicine for full access: SubscribeLogin for existing subscribers Additional access options: * Use a document delivery service * Rent this article from DeepDyve * Login via Athens * Purchase a site license * Institutional access * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data - Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion
- Nat Med 17(2):175-178 (2011)
Nature Medicine | Brief Communication Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion * Ryuichiro Atarashi1, 2 Contact Ryuichiro Atarashi Search for this author in: * NPG journals * PubMed * Google Scholar * Katsuya Satoh1 Search for this author in: * NPG journals * PubMed * Google Scholar * Kazunori Sano1, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Takayuki Fuse1 Search for this author in: * NPG journals * PubMed * Google Scholar * Naohiro Yamaguchi1 Search for this author in: * NPG journals * PubMed * Google Scholar * Daisuke Ishibashi1 Search for this author in: * NPG journals * PubMed * Google Scholar * Takehiro Matsubara1 Search for this author in: * NPG journals * PubMed * Google Scholar * Takehiro Nakagaki1 Search for this author in: * NPG journals * PubMed * Google Scholar * Hitoki Yamanaka4 Search for this author in: * NPG journals * PubMed * Google Scholar * Susumu Shirabe5 Search for this author in: * NPG journals * PubMed * Google Scholar * Masahito Yamada6 Search for this author in: * NPG journals * PubMed * Google Scholar * Hidehiro Mizusawa7 Search for this author in: * NPG journals * PubMed * Google Scholar * Tetsuyuki Kitamoto8 Search for this author in: * NPG journals * PubMed * Google Scholar * Genevieve Klug9 Search for this author in: * NPG journals * PubMed * Google Scholar * Amelia McGlade9 Search for this author in: * NPG journals * PubMed * Google Scholar * Steven J Collins9 Search for this author in: * NPG journals * PubMed * Google Scholar * Noriyuki Nishida1, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:175–178Year published:(2011)DOI:doi:10.1038/nm.2294Received02 July 2010Accepted16 December 2010Published online30 January 2011 Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The development of technologies for the in vitro amplification of abnormal conformations of prion protein (PrPSc) has generated the potential for sensitive detection of prions. Here we developed a new PrPSc amplification assay, called real-time quaking-induced conversion (RT-QUIC), which allows the detection of ≥1 fg of PrPSc in diluted Creutzfeldt-Jakob disease (CJD) brain homogenate. Moreover, we assessed the technique first in a series of Japanese subjects and then in a blind study of 30 cerebrospinal fluid specimens from Australia, which achieved greater than 80% sensitivity and 100% specificity. These findings indicate the promising enhanced diagnostic capacity of RT-QUIC in the antemortem evaluation of suspected CJD. View full text Author information * Author information * Supplementary information Affiliations * Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. * Ryuichiro Atarashi, * Katsuya Satoh, * Kazunori Sano, * Takayuki Fuse, * Naohiro Yamaguchi, * Daisuke Ishibashi, * Takehiro Matsubara, * Takehiro Nakagaki & * Noriyuki Nishida * Nagasaki University Research Centre for Genomic Instability and Carcinogenesis, Nagasaki, Japan. * Ryuichiro Atarashi * Global Centers of Excellence Program, Nagasaki University, Nagasaki, Japan. * Kazunori Sano & * Noriyuki Nishida * Division of Comparative Medicine, Center for Frontier Life Sciences, Nagasaki University, Nagasaki, Japan. * Hitoki Yamanaka * Organization of Rural Medicine and Residency Education, Nagasaki University Hospital, Nagasaki, Japan. * Susumu Shirabe * Department of Neurology and Neurobiology of Aging, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan. * Masahito Yamada * Department of Neurology and Neurological Science, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan. * Hidehiro Mizusawa * Division of Creutzfeldt-Jakob Disease Science and Technology, Department of Prion Research, Tohoku University Graduate School of Medicine, Sendai, Japan. * Tetsuyuki Kitamoto * Department of Pathology, Australian National Creutzfeldt-Jakob Disease Registry, University of Melbourne, Melbourne, Australia. * Genevieve Klug, * Amelia McGlade & * Steven J Collins Contributions R.A. designed the project, performed experiments and wrote the manuscript. K. Satoh, K. Sano, T.F., N.Y., D.I., T.M., T.N. and H.Y. performed experiments. K. Satoh, S.S., M.Y., H.M., T.K., G.K., A.M. and S.J.C. contributed to the collection of human specimens and provided information about subjects. N.N. supervised the project. K. Satoh, K. Sano, A.M., S.J.C. and N.N. helped with the editing of the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * Ryuichiro Atarashi Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (938K) Supplementary Figures 1–3 and Supplementary Tables 1–4 and Supplementary Methods Additional data - The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance
- Nat Med 17(2):179-188 (2011)
Nature Medicine | Article The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance * Bolormaa Vandanmagsar1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Yun-Hee Youm1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Anthony Ravussin1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Jose E Galgani2, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Krisztian Stadler2, 4 Search for this author in: * NPG journals * PubMed * Google Scholar * Randall L Mynatt2 Search for this author in: * NPG journals * PubMed * Google Scholar * Eric Ravussin2, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Jacqueline M Stephens5 Search for this author in: * NPG journals * PubMed * Google Scholar * Vishwa Deep Dixit1, 2 Contact Vishwa Deep Dixit Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:179–188Year published:(2011)DOI:doi:10.1038/nm.2279Received18 May 2010Accepted18 November 2010Published online09 January 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The emergence of chronic inflammation during obesity in the absence of overt infection or well-defined autoimmune processes is a puzzling phenomenon. The Nod-like receptor (NLR) family of innate immune cell sensors, such as the nucleotide-binding domain, leucine-rich–containing family, pyrin domain–containing-3 (Nlrp3, but also known as Nalp3 or cryopyrin) inflammasome are implicated in recognizing certain nonmicrobial originated 'danger signals' leading to caspase-1 activation and subsequent interleukin-1β (IL-1β) and IL-18 secretion. We show that calorie restriction and exercise-mediated weight loss in obese individuals with type 2 diabetes is associated with a reduction in adipose tissue expression of Nlrp3 as well as with decreased inflammation and improved insulin sensitivity. We further found that the Nlrp3 inflammasome senses lipotoxicity-associated increases in intracellular ceramide to induce caspase-1 cleavage in macrophages and adipose tissue. Ablation of Nl! rp3 in mice prevents obesity-induced inflammasome activation in fat depots and liver as well as enhances insulin signaling. Furthermore, elimination of Nlrp3 in obese mice reduces IL-18 and adipose tissue interferon-γ (IFN-γ) expression, increases naive T cell numbers and reduces effector T cell numbers in adipose tissue. Collectively, these data establish that the Nlrp3 inflammasome senses obesity-associated danger signals and contributes to obesity-induced inflammation and insulin resistance. View full text Figures at a glance * Figure 1: Reduction of Nlrp3 and IL-1β expression is associated with improvement of insulin sensitivity. (,) Positive correlation of the visceral fat mRNA expression of Il1b () and Nlrp3 () with body weight of C57BL/6 mice (n = 32); Pearson's correlations are r = 0.364, P = 0.0178 for Il1b and r = 0.672, P < 0.0001 for Nlrp3, respectively. (–) Il1b (), Nlrp3 () and Pycard () mRNA in visceral and subcutaneous adipose tissue from ad libitum chow-fed control (AL) and 40% calorie-restricted (CR) 12-month-old mice, n = 6; *P < 0.01, **P < 0.005. (,) Representative H&E staining showing adipocyte size in visceral () and subcutaneous () fat tissue from ad libitum–fed control and calorie-restricted 12-month-old C57BL/6 mice. () IL1B (left), NLRP3 (middle) and PYCARD (right) gene expression, as examined by quantitative RT-PCR (qRT-PCR) in human SAT in obese individuals with T2DM before and after 1-year weight loss. *P = 0.01, **P = 0.001. () Positive correlation of changes in gene expression of IL1B and NLRP3 in human abdominal subcutaneous fat with changes in fasting glucose level f! rom baseline to 1 year after intervention; Pearson's correlations are r = 0.53, P = 0.12 for IL1B and r = 0.69, P = 0.03 for NLRP3. Relative gene expression levels are depicted as means ± s.e.m. n = 10. * Figure 2: Elimination of Nlrp3 expression prevents obesity-induced caspase-1 cleavage and IL-1β and IL-18 activation. () Immunofluorescence staining of epididymal fat (eFat) tissue sections stained with antibodies against F4/80 (red) and Nlrp3 (green). Merge of images with nuclear staining with DAPI shows colocalization of Nlrp3 with ATM (yellow arrowheads). Negative control staining with antibody to Nlrp3 together with antibody to F4/80 in adipose tissue of Nlrp3−/− mice shows reduced Nlrp3-specific immunostaining. (,) The qRT-PCR analysis of Nlrp3 () and Pycard () mRNA in purified ATM and SVF cells derived from SAT and VAT of 6-month-old DIO mice, and mature 3T3-L1 adipocytes (Adip). () Immunoblot analysis showing the kinetics of caspase-1 (p20) cleavage and active IL-1β (p17) accumulation in adipose tissue of mice at various stages of diet-induced obesity. The age, in months, of the mice at the time of tissue harvesting is indicated. () Western blot analysis of activated caspase-1 (p20) in VAT, SAT and liver tissues from 9-month-old DIO-WT (C57BL/6) and DIO-Nlrp3−/− mice. Result! s shown are representative of three independent experiments. MW, molecular weight. () Western blot analysis of activated caspase-1 (p20) in kidney from 9-month-old DIO-WT and DIO-Nlrp3−/− mice. () Western blot analysis of IL-1β activation in adipose tissue of 6- and 7-month-old DIO mice. () Serum IL-18 concentration in age-matched WT and Nlrp3−/− mice fed normal chow or 60% HFD, starting at 2 months of age for all groups, for 4 months and 7 months. All data are presented as means ± s.e.m., n = 6–10 mice; *P < 0.05. * Figure 3: The Nlrp3 inflammasome regulates insulin sensitivity and steatohepatitis in obesity. () Insulin tolerance test (ITT) and glucose tolerance test (GTT) in male WT and Nlrp3−/− DIO mice fed 60% HFD for 6 weeks. (,) The ITT and GTT in 6-month-old () and 9-month-old () WT and Nlrp3−/− DIO mice showing the area under the curve (green for WT and blue for Nlrp3−/−) with trend lines (n = 5–7 per group). () The total area under the curve (AUC) for ITT in 6- (left) and 9-month-old (right) WT and Nlrp3−/− DIO mice. (*P < 0.01). () Adipocyte size in visceral fat tissue from 8-month-old DIO-WT (Nlrp3+/+) and Nlrp3−/− mice (H&E staining). Representative micrographs and the quantified results are shown. n = 6; *P < 0.001. () Akt, IRS-1 and Erk-1/2 signaling in vivo, as determined by western blotting for phosphorylation, in VAT, SAT, liver and muscle from the 8-month-old DIO Nlrp3+/+ and DIO Nlrp3−/− mice. The Ser322/338 antibody detects phosphorylation at Ser332 and Ser336. () H&E staining of sections from liver of 9-month-old DIO WT and Nlrp3−/�! �� mice. () Fatty acid oxidation and fatty acid synthesis gene expression, as examined by qRT-PCR in the liver of 9-month-old DIO WT and Nlrp3−/− mice. mRNA expression was normalized to glyceraldehyde 3-phosphate dehydrogenase (Gapdh) expression and is shown as fold change (ΔΔCt) with the values for WT arbitrarily set to 1. All data are presented as means ± s.e.m. (n = 5), *P < 0.05, **P < 0.001. * Figure 4: Nlrp3 senses ceramide to induce IL-1β and regulates adipose tissue macrophage activation in obesity. () The cell extracts of LPS-primed bone marrow–derived macrophages (BMDMs) from Nlrp3+/+ and Nlrp3−/− mice (n = 6–9) were analyzed for caspase-1 active form by western blotting as indicated. () The BMDM cells were stimulated with LPS and C2 ceramide (0.1 mM), and cell supernatants were analyzed for IL-1β. Results are representative of three separate experiments. () Epididymal adipose tissue explants from 9-month-old Nlrp3+/+ and Nlrp3−/− DIO mice were cultured for 24 h in the presence of LPS, C2 ceramide or both. Caspase-1 activation was determined by immunoblot analysis. () M1- and M2-associated gene expression, as examined by qRT-PCR in macrophages originating from VAT and SAT of 9-month-old DIO-Nlrp3+/+ and DIO-Nlrp3−/− mice. The mRNA expression was normalized to glyceraldehydes 3-phosphate dehydrogenase (Gapdh) and shown as fold change (ΔΔCt) with the values for WT arbitrarily set to 1. All data are presented as means ± s.e.m., *P < 0.01, **P < 0.001. * Figure 5: Ablation of the Nlrp3 inflammasome reduces adipose tissue effector T cells without affecting Treg cells in visceral fat of obese mice. () FACS plot of SVF cells isolated from VAT of 9-month-old WT and Nlrp3−/− obese mice. The dot plots depict FSC and SSC (left) and the sequential gating strategy for analysis of ATMs and T cells. Gate 1 (top) of larger cells shows the presence of F4/80+ cells (histogram) and expression of macrophage markers CD206 and CD11c on ATMs. Gating of smaller cells (gate 2, bottom) reveals the absence of ATMs in this population of SVF. () The gate 2 (lymphoid gate) showing CD4+ and CD8+ T cells in SVF. CD4+ and CD8+ cells were evaluated for naive T cells (CD62L+CD44−, blue boxes) and effector memory (CD62L−CD44+, red boxes) CD4+ and CD8+ T cells. The FACS analysis was repeated in three independent pooled SVF fractions from a total of 12–14 mice and percent gated cell frequencies are indicated in each representative dot plot. () Gated percentage and absolute numbers (in million cells) of naive (CD62L+CD44−) and effector memory (CD62L−CD44+) CD4+ and CD8+ T cells. () Numbe! r of stromal vascular cells per gram of fat tissue (n = 4–6) in 3- and 9-month-old WT and Nlrp3−/− DIO mice. () Representative FACS plots showing CD4+CD25+Foxp3+ T regulatory cells in VAT of 9-month-old WT and Nlrp3−/− DIO mice. () Gated percentage of Treg cells in VAT and SAT of 9-month-old WT and Nlrp3−/− DIO mice (n = 6 per group). All data are presented as means ± s.e.m., *P < 0.05. * Figure 6: Elimination of the Nlrp3 inflammasome reduces obesity-induced macrophage–mediated T cell activation in adipose tissue. () Representative FACS plots of SVF cells from SAT of 6- and 9-month-old WT and Nlrp3−/− DIO mice stained with CD206 and CD11c. () The gated percentage of CD11c−CD206+ M2 cells in SAT of 9-month-old obese mice. () Dot plots showing naive (CD62L+CD44−, blue boxes) and effector memory (CD62L−CD44+, red boxes) CD4+ and CD8+ T cells in SVF from 9-month-old obese WT and Nlrp3−/− mice. The percent gated cell frequencies are indicated in each representative dot plot () Gated percentage and absolute numbers (in million cells) of naive (CD62L+CD44−) and effector memory (CD62L−CD44+) CD4+ and CD8+ T cells. () mRNA level of the TH1 cytokine Ifnγ in VAT and SAT of 9-month-old obese WT and Nlrp3−/− mice as determined by qRT–PCR. () IFN-γ (19-kDa) in VAT of 9-month-old WT and Nlrp3−/− DIO mice, as examined by western blotting. () IP10 and MCP-1 levels in the serum of 9-month-old lean WT and obese WT and Nlrp3−/− mice (n = 5). The data shown are means ± s! .e.m., *P < 0.05. () Hypothetical model of Nlrp3 inflammasome activation in obesity. In the absence of danger signals in the healthy lean state, tissue-resident macrophages and T cells may participate in the maintenance of adipose tissue function. In obesity, the Nlrp3 inflammasome senses the obesity-associated danger signals such as ceramides leading to caspase-1 autoactivation and IL-1β and IL-18 production from ATMs. Secondary signals from activated ATMs to effector adipose T cells (defined as CD44+CD62L−) sustain the reciprocal proinflammatory feed-forward cascade during obesity, leading to insulin resistance. ROS, reactive oxygen species. Author information * Abstract * Author information * Supplementary information Affiliations * Laboratory of Neuroendocrine-Immunology, Louisiana State University, Baton Rouge, Louisiana, USA. * Bolormaa Vandanmagsar, * Yun-Hee Youm, * Anthony Ravussin & * Vishwa Deep Dixit * Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, Louisiana, USA. * Bolormaa Vandanmagsar, * Yun-Hee Youm, * Anthony Ravussin, * Jose E Galgani, * Krisztian Stadler, * Randall L Mynatt, * Eric Ravussin & * Vishwa Deep Dixit * Human Physiology Laboratory, Louisiana State University, Baton Rouge, Louisiana, USA. * Jose E Galgani & * Eric Ravussin * Oxidative Stress and Disease Laboratory, Louisiana State University, Baton Rouge, Louisiana, USA. * Krisztian Stadler * Department of Biological Sciences, Louisiana State University System, Baton Rouge, Louisiana, USA. * Jacqueline M Stephens Contributions B.V. performed real-time PCRs, flow cytometry assays, adipose tissue macrophage selections, some western blots, ITT, GTT and cytokine assays, managed the transgenic animal colony and participated in experimental design, data analysis and manuscript preparation. Y.-H.Y. performed all caspase-1 and IL1β western blots, adipose and liver histologies, and macrophage culture experiments and analyzed the data. A.R. Performed body composition analysis, tissue collections, lipid analysis, animal husbandry, genotyping, ITT, GTT and adipocyte size measurement. J.E.G. and E.R. designed and supervised the human studies, analyzed the glucose and insulin sensitivity data in obese T2DM subjects and discussed the hypotheses. K.S. performed the caspase-1 western blot in the kidneys of HFD fed WT and Nlrp3 null mice. R.L.M. participated in standardizing the ITT and GTT assays and advised on the design of experiments for liver fatty acid synthesis and oxidation gene expression. J.M.S. performe! d the western blots for some of the insulin-signaling experiments, helped with data interpretation, discussed the hypotheses and participated in manuscript preparation. V.D.D. conceived the project, designed the experiments, performed some of the cytokine assays and flow cytometry, helped with data interpretation, participated in data analysis, directed the project and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * Vishwa Deep Dixit Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (376K) Supplementary Figures 1–7 and Supplementary Table 1 Additional data - A multistage tuberculosis vaccine that confers efficient protection before and after exposure
- Nat Med 17(2):189-194 (2011)
Nature Medicine | Article A multistage tuberculosis vaccine that confers efficient protection before and after exposure * Claus Aagaard1, 6 Contact Claus Aagaard Search for this author in: * NPG journals * PubMed * Google Scholar * Truc Hoang1, 6 Search for this author in: * NPG journals * PubMed * Google Scholar * Jes Dietrich1 Search for this author in: * NPG journals * PubMed * Google Scholar * Pere-Joan Cardona2 Search for this author in: * NPG journals * PubMed * Google Scholar * Angelo Izzo3 Search for this author in: * NPG journals * PubMed * Google Scholar * Gregory Dolganov4 Search for this author in: * NPG journals * PubMed * Google Scholar * Gary K Schoolnik4 Search for this author in: * NPG journals * PubMed * Google Scholar * Joseph P Cassidy5 Search for this author in: * NPG journals * PubMed * Google Scholar * Rolf Billeskov1 Search for this author in: * NPG journals * PubMed * Google Scholar * Peter Andersen1 Contact Peter Andersen Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorsJournal name:Nature MedicineVolume: 17,Pages:189–194Year published:(2011)DOI:doi:10.1038/nm.2285Received12 October 2010Accepted07 December 2010Published online23 January 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg All tuberculosis vaccines currently in clinical trials are designed as prophylactic vaccines based on early expressed antigens. We have developed a multistage vaccination strategy in which the early antigens Ag85B and 6-kDa early secretory antigenic target (ESAT-6) are combined with the latency-associated protein Rv2660c (H56 vaccine). In CB6F1 mice we show that Rv2660c is stably expressed in late stages of infection despite an overall reduced transcription. The H56 vaccine promotes a T cell response against all protein components that is characterized by a high proportion of polyfunctional CD4+ T cells. In three different pre‐exposure mouse models, H56 confers protective immunity characterized by a more efficient containment of late-stage infection than the Ag85B-ESAT6 vaccine (H1) and BCG. In two mouse models of latent tuberculosis, we show that H56 vaccination after exposure is able to control reactivation and significantly lower the bacterial load compared to adjuvant ! control mice. View full text Figures at a glance * Figure 1: Immunogenicity and protective efficacy of H56 and its components. () Purified Ag85B, ESAT-6, Rv2660c and H56 were visualized in Coomassie-stained gels. Ag85B, lane 1; ESAT-6, lane 2; Rv2660c, lane 3 and H56, lane 4. MW, molecular weight. () Protective efficacies in H56/CAF01 vaccinated CB6F1 mice challenged with Mtb (Erdman). One BCG vaccinated group is included as positive control. The differences between vaccinated and nonvaccinated mice 6 weeks after challenge are shown from one of two experiments. () Antigen-specific IFN-γ released from splenocytes purified from Ag85B-, ESAT-6–, Rv2660c- or H56-immunized CB6F1 mice (n = 3 per group) after in vitro stimulation with Ag85B, ESAT-6 or Rv2660c. Results are shown from one of three experiments. () Bacterial numbers in the lung of individual mice (CB6F1) 6 weeks after challenge (n = 6 per group). Data from one of three experiments are shown. () The development of the infection in mice (C57BL/6) immunized with BCG, H56 or saline and challenged with Mtb (H37Rv) was followed by enumerating the! bacilli in the lung of individual mice (n = 6) over a period of 24 weeks. In and , one-way analysis of variance (ANOVA) was used for group comparisons. *P < 0.05; **P < 0.01; ***P < 0.001 compared to the adjuvant control group or BCG (}). Data are means ± s.e.m. * Figure 2: Immune responses and vaccine efficacy of H56 compared to H1. () Antigen-specific IFN-γ release from in vitro–stimulated splenocytes isolated from mice (CB6F1) vaccinated with H1 or H56. Representative results from one of three experiments are shown. *P < 0.05, one-way ANOVA. () IFN-γ released from splenocytes isolated after challenge with Mtb (Erdman) from groups of CB6F1 mice vaccinated with CAF01 adjuvant, BCG, H1 or H56 and stimulated in vitro with the indicated antigens. Results from one of two independent experiments are shown. () The frequency of Ag85B-specific CD4+ T cells (CD44high) producing IFN-γ, TNF-α or IL-2 measured in cells isolated from perfused lungs from mice immunized with CAF01, BCG, H1 or H56. The cytokine profile in individual cells was measured by multicolor flow cytometry by gating for lymphocytes and CD4+ T cells. All possible combinations of cytokine expression were tabulated, and, after subtracting the background (non‐stimulated samples), the results for the seven combinations expressing at least one! of the cytokines are shown. Each of the panels – show results from the same experiment. Two independent experiments were performed. () The bacterial load (CFUs) in the lungs of individual mice. The results are pooled values from two experiments, and each time point represents results from 10–12 mice per group. *P < 0.05; **P < 0.01 using one-way ANOVA. Data are means ± s.e.m. * Figure 3: Evaluation of H56 as a BCG booster. (,) Bacterial numbers (CFUs) 6 () and 24 () weeks after challenge with Mtb (Erdman) in the lung (n = 6 per group) of mice (CB6F1) vaccinated with BCG and boosted twice with either H1 or H56. Representative data from one of two experiments are shown as log10 CFU. ***P < 0.001; **P < 0.01; *P < 0.05, one-way ANOVA with Tukey's post test. Data are means ± s.e.m. * Figure 4: Vaccination with H56 after exposure. () The model used for evaluation of the H56 vaccine after exposure. Mice infected with Mtb (Erdman) were treated with antibiotic for 6 weeks (shaded area). After treatment, mice were killed and bacteria were enumerated in the lungs at the indicated time points (n = 6). The arrows indicate the vaccination time points used in , and . Log10 CFUs are given as mean values ± s.e.m. () IFN-γ released from PBMCs isolated 35 weeks after infection with Mtb (Erdman) from mice vaccinated at weeks 10, 13 and 16 and from nonvaccinated mice (n = 16 per group, pooled PBMCs). PBMCs were stimulated in vitro with Ag85B, ESAT-6 or Rv2660c. () At the same time point, cytokine profiles of antigen-specific CD4+ T cells were measured in Ag85B-stimulated splenocytes by flow cytometry as described in Figure 2c. () The protective efficacy of H56 was measured in two different laboratories. Experiments 1–3 were done in CB6F1 mice at Statens Serum Institut and experiments 4–6 were done in C57BL/6 m! ice at Unitat de Tuberculosi Experimental. In experiments 1–3, bacteria were enumerated in the lungs of individual mice 35 weeks (exp. 1 and 3) and 43 weeks (exp. 2) after challenge. In experiments 1 and 2, mice received two vaccinations, and in experiment 3, three. In experiments 4–6, mice received two vaccinations, and the bacterial load was measured 23 weeks after infection. The CFU values are shown as scattered plots with the median indicated (n = 12–16 per group in each experiment). We used the Mann-Whitney U test for comparison among groups. *P < 0.05; **P < 0.01; ***P < 0.001. Data are means ± s.e.m. Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Claus Aagaard & * Truc Hoang Affiliations * Department of Infectious Disease Immunology, Statens Serum Institut, Copenhagen, Denmark. * Claus Aagaard, * Truc Hoang, * Jes Dietrich, * Rolf Billeskov & * Peter Andersen * Unitat de Tuberculosi Experimental, Institut per a la Investigació en Ciències de la Salut Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona, Catalonia, Spain. * Pere-Joan Cardona * Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA. * Angelo Izzo * Department of Microbiology and Immunology, Stanford University School of Medicine, California, USA. * Gregory Dolganov & * Gary K Schoolnik * Veterinary Sciences Centre, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin, Ireland. * Joseph P Cassidy Contributions C.A. conceived of the study, produced H56, conducted pre‐exposure vaccine studies and prepared the manuscript. T.H. developed the CB6F1 latency model and conducted latency studies in this model. J.D. conducted the BCG boost studies. P.-J.C. developed the C57BL/6 latency model and conducted latency studies in this model. A.I. conducted pre‐exposure vaccine studies. G.D. designed and performed gene expression analyses. G.K.S. designed and performed gene expression analyses. J.P.C. performed histological evaluation of lung specimens. R.B. contributed to the latency vaccine studies. P.A. conceived of the study and prepared the manuscript. All authors discussed the results and commented on the manuscript at all stages. Competing financial interests C.A. and P.A. are co-inventors on a patent application to the Danish patent office covering the use of H56 as a vaccine. All rights have been assigned to Statens Serum Institut, a Danish not-for-profit governmental institute. Corresponding authors Correspondence to: * Claus Aagaard or * Peter Andersen Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (315K) Supplementary Figures 1 and 2 Additional data - Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes
- Nat Med 17(2):195-199 (2011)
Nature Medicine | Letter Severe pandemic 2009 H1N1 influenza disease due to pathogenic immune complexes * Ana Clara Monsalvo1, 9 Search for this author in: * NPG journals * PubMed * Google Scholar * Juan P Batalle1, 9 Search for this author in: * NPG journals * PubMed * Google Scholar * M Florencia Lopez1, 9 Search for this author in: * NPG journals * PubMed * Google Scholar * Jens C Krause2 Search for this author in: * NPG journals * PubMed * Google Scholar * Jennifer Klemenc2 Search for this author in: * NPG journals * PubMed * Google Scholar * Johanna Zea Hernandez1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Bernardo Maskin3 Search for this author in: * NPG journals * PubMed * Google Scholar * Jimena Bugna1 Search for this author in: * NPG journals * PubMed * Google Scholar * Carlos Rubinstein4 Search for this author in: * NPG journals * PubMed * Google Scholar * Leandro Aguilar4 Search for this author in: * NPG journals * PubMed * Google Scholar * Liliana Dalurzo5 Search for this author in: * NPG journals * PubMed * Google Scholar * Romina Libster1 Search for this author in: * NPG journals * PubMed * Google Scholar * Vilma Savy6 Search for this author in: * NPG journals * PubMed * Google Scholar * Elsa Baumeister6 Search for this author in: * NPG journals * PubMed * Google Scholar * Liliana Aguilar3 Search for this author in: * NPG journals * PubMed * Google Scholar * Graciela Cabral3 Search for this author in: * NPG journals * PubMed * Google Scholar * Julia Font3 Search for this author in: * NPG journals * PubMed * Google Scholar * Liliana Solari3 Search for this author in: * NPG journals * PubMed * Google Scholar * Kevin P Weller2 Search for this author in: * NPG journals * PubMed * Google Scholar * Joyce Johnson7 Search for this author in: * NPG journals * PubMed * Google Scholar * Marcela Echavarria8 Search for this author in: * NPG journals * PubMed * Google Scholar * Kathryn M Edwards2 Search for this author in: * NPG journals * PubMed * Google Scholar * James D Chappell7 Search for this author in: * NPG journals * PubMed * Google Scholar * James E Crowe Jr2 Search for this author in: * NPG journals * PubMed * Google Scholar * John V Williams2 Search for this author in: * NPG journals * PubMed * Google Scholar * Guillermina A Melendi1, 2 Contact Guillermina A Melendi Search for this author in: * NPG journals * PubMed * Google Scholar * Fernando P Polack1, 2 Contact Fernando P Polack Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorsJournal name:Nature MedicineVolume: 17,Pages:195–199Year published:(2011)DOI:doi:10.1038/nm.2262Received19 May 2010Accepted19 October 2010Published online05 December 2010 Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Pandemic influenza viruses often cause severe disease in middle-aged adults without preexisting comorbidities. The mechanism of illness associated with severe disease in this age group is not well understood1, 2, 3, 4, 5, 6, 7, 8, 9, 10. Here we find preexisting serum antibodies that cross-react with, but do not protect against, 2009 H1N1 influenza virus in middle-aged adults. Nonprotective antibody is associated with immune complex–mediated disease after infection. We detected high titers of serum antibody of low avidity for H1-2009 antigen, and low-avidity pulmonary immune complexes against the same protein, in severely ill individuals. Moreover, C4d deposition—a marker of complement activation mediated by immune complexes—was present in lung sections of fatal cases. Archived lung sections from middle-aged adults with confirmed fatal influenza 1957 H2N2 infection revealed a similar mechanism of illness. These observations provide a previously unknown biological mecha! nism for the unusual age distribution of severe cases during influenza pandemics. View full text Figures at a glance * Figure 1: Histopathology and virus titers in 2009 H1N1 disease. () Pulmonary histopathology in representative lung sections of fatal 2009 H1N1 and seasonal H1N1 virus-infected humans (H&E). () Detection of 2009 H1N1 and seasonal H1N1 influenza viruses. Scale bar, 100 μm. The boxes are details of pulmonary edema (2009 H1N1, H&E), peribronchiolar mononuclear cell infiltration (seasonal H1N1, H&E), and virus-infected cells (anti-H1N1 stains). Scale bars, 100 μm. () 2009 H1N1 RT-PCR Ct values in nasopharyngeal secretions of fatal, intensive care unit (ICU) and ambulatory cases; there were no significant differences. () 2009 H1N1 RT-PCR Ct values by time of symptoms before nasopharyngeal sampling. * Figure 2: Inflammation in influenza A 2009 H1N1 disease. (–) IL-8 (), TNF-α (), IL-6 () and IL-1β () responses in respiratory secretions of individuals infected with 2009 H1N1 influenza virus (TA, tracheal aspirates; NP, nasopharyngeal secretions) or seasonal influenza viruses (H1N1 and H3N2). Comparisons for nasopharyngeal secretions: IL-8, P = 0.01; all other cytokines, not significant. (–) IL-1β (), IL-6 (), IL-10 () and TNF-α () production by human monocytes incubated with a dose range of recombinant protein H1-1918, H1-1999, H1-2009, avian H5, hMPV F and a control monoclonal human IgG against the Sa antigenic site of influenza hemagglutinin (2D1). Representative of three independent experiments. () Individual TLR activation in HEK293 cells. Dose: 5 μg of H1-1918, H1-1999, H1-2009, hMPV F and no ligand. Representative of two independent experiments. Error bars represent s.e.m. * Figure 3: Lymphopenia in influenza A 2009 H1N1 disease. (–) CD3+ (), CD4+ () and CD8+ () T lymphocyte counts in fatal and ICU cases of 2009 H1N1 influenza A virus on admission. (,) Immunohistochemistry for CD3+ T lymphocytes (red arrows) (), CD8+ T lymphocytes (yellow arrows) () in representative lung sections of fatal influenza 2009 H1N1 and seasonal infection. Positive controls are archived sections from different organs in unrelated subjects provided for every antibody. Negative controls are lung sections from an individual who died from a nonpulmonary disease (scale bars, 100 μm). * Figure 4: Immune complex–mediated disease in 2009 H1N1 influenza infection. () Serum IgG endpoint titers against hemagglutinin proteins by immunoassay in infants (mean age (range), 7.6 months (6.1–11.8); n = 10), middle-aged adults (n = 16) and elderly (n = 12). () Protein-specific avidity of IgG after 6–9 M urea wash in naive adults (age range, 25–43 years old) and elderly (age range, 75–97 years old); P < 0.05. () Microneutralization titers for infants, adults and the elderly; P < 0.05 for elderly versus both groups. () Endpoint titer of serum IgG against H1-2009 antigens by immunoassay in adults with severe (n = 12) versus mild (n = 11) disease, P < 0.05. () H1-2009–specific avidity of IgG after 7–9 M urea wash in infected adults with severe (n = 12) versus mild (n = 11) pandemic disease, P < 0.05. () H1-2009–specific avidity of IgG from immune complexes after 8 M urea wash in infected adults with severe (n = 14) versus mild (n = 11) pandemic disease, P < 0.05. () C4d detection in representative slides from lung sections of two of s! ix middle-aged fatal cases of 2009 H1N1 influenza showing extensive peribronchiolar immune complex–mediated complement activation. There is trace C4d deposition in representative lung section of fatal seasonal H1N1 virus from an elderly woman. Positive control, C4d deposition in a kidney from an adult with immune complex–mediated glomerulonephritis; negative control, a lung section from an adult with solid tumor (scale bars, 100 μm). () Serum complement C3 concentrations in adults who required intensive care or hospital ward admission, and infants infected with H1N1 2009 influenza A virus, adults infected with seasonal influenza virus and adults with pulmonary diseases other than influenza in the ICU and floor; P = 0.036. () C4d detection in representative slides from lung sections of two fatal 1957 H2N2 influenza cases showing extensive peribronchiolar deposition of complement cleavage product. Negative control from an archived sample with no infectious pulmonary dise! ase. The box in the microphotograph is a detail of lung comple! ment deposition. Author information * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Ana Clara Monsalvo, * Juan P Batalle & * M Florencia Lopez Affiliations * Fundacion INFANT, Buenos Aires, Argentina. * Ana Clara Monsalvo, * Juan P Batalle, * M Florencia Lopez, * Johanna Zea Hernandez, * Jimena Bugna, * Romina Libster, * Guillermina A Melendi & * Fernando P Polack * Department of Pediatrics, Vanderbilt University, Nashville, Tennessee, USA. * Jens C Krause, * Jennifer Klemenc, * Johanna Zea Hernandez, * Kevin P Weller, * Kathryn M Edwards, * James E Crowe Jr, * John V Williams, * Guillermina A Melendi & * Fernando P Polack * Hospital Nacional Profesor Alejandro Posadas, Buenos Aires, Argentina. * Bernardo Maskin, * Liliana Aguilar, * Graciela Cabral, * Julia Font & * Liliana Solari * Hospital Dr. Federico Abete, Malvinas Argentinas, Buenos Aires, Argentina. * Carlos Rubinstein & * Leandro Aguilar * Hospital Italiano, Buenos Aires, Argentina. * Liliana Dalurzo * Administración Nacional de Laboratorios e Institutos de Salud Dr. Carlos G. Malbran, Buenos Aires, Argentina. * Vilma Savy & * Elsa Baumeister * Department of Pathology, Vanderbilt University, Nashville, Tennessee, USA. * Joyce Johnson & * James D Chappell * Department of Microbiology, Centro de Educación Médica e Investigaciones Clínicas, Buenos Aires, Argentina. * Marcela Echavarria Contributions F.P.P., G.A.M., K.M.E., J.D.C., J.E.C. Jr., J.V.W., A.C.M., J.P.B. and M.F.L. designed the project. A.C.M., J.P.B., M.F.L., J.Z.H., B.M., L.D., K.P.W., J.V.W., G.A.M. and F.P.P. performed experiments. J.C.K., J.K., J.B., C.R., Le. A., L.D., R.L., V.S., E.B., Li. A., G.C., J.F., L.S., J.J., M.E., J.E.C. Jr. and J.V.W. developed or provided key reagents or contributed samples. F.P.P. supervised the project. A.C.M., J.P.B., M.F.L., K.M.E., J.D.C., G.A.M. and F.P.P. wrote the paper. Competing financial interests The authors declare no competing financial interests. Corresponding authors Correspondence to: * Fernando P Polack or * Guillermina A Melendi Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (636K) Supplementary Figures 1–6 Additional data - Brown adipose tissue activity controls triglyceride clearance
- Nat Med 17(2):200-205 (2011)
Nature Medicine | Letter Brown adipose tissue activity controls triglyceride clearance * Alexander Bartelt1 Contact Alexander Bartelt Search for this author in: * NPG journals * PubMed * Google Scholar * Oliver T Bruns2 Search for this author in: * NPG journals * PubMed * Google Scholar * Rudolph Reimer2 Search for this author in: * NPG journals * PubMed * Google Scholar * Heinz Hohenberg2 Search for this author in: * NPG journals * PubMed * Google Scholar * Harald Ittrich3 Search for this author in: * NPG journals * PubMed * Google Scholar * Kersten Peldschus3 Search for this author in: * NPG journals * PubMed * Google Scholar * Michael G Kaul3 Search for this author in: * NPG journals * PubMed * Google Scholar * Ulrich I Tromsdorf4 Search for this author in: * NPG journals * PubMed * Google Scholar * Horst Weller4 Search for this author in: * NPG journals * PubMed * Google Scholar * Christian Waurisch5 Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander Eychmüller5 Search for this author in: * NPG journals * PubMed * Google Scholar * Philip L S M Gordts6 Search for this author in: * NPG journals * PubMed * Google Scholar * Franz Rinninger7 Search for this author in: * NPG journals * PubMed * Google Scholar * Karoline Bruegelmann1 Search for this author in: * NPG journals * PubMed * Google Scholar * Barbara Freund1 Search for this author in: * NPG journals * PubMed * Google Scholar * Peter Nielsen1 Search for this author in: * NPG journals * PubMed * Google Scholar * Martin Merkel1, 8 Search for this author in: * NPG journals * PubMed * Google Scholar * Joerg Heeren1 Contact Joerg Heeren Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorsJournal name:Nature MedicineVolume: 17,Pages:200–205Year published:(2011)DOI:doi:10.1038/nm.2297Received12 October 2010Accepted22 December 2010Published online23 January 2011 Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Brown adipose tissue (BAT) burns fatty acids for heat production to defend the body against cold1, 2 and has recently been shown to be present in humans3, 4, 5. Triglyceride-rich lipoproteins (TRLs) transport lipids in the bloodstream, where the fatty acid moieties are liberated by the action of lipoprotein lipase (LPL)6. Peripheral organs such as muscle and adipose tissue take up the fatty acids, whereas the remaining cholesterol-rich remnant particles are cleared by the liver6. Elevated plasma triglyceride concentrations and prolonged circulation of cholesterol-rich remnants, especially in diabetic dyslipidemia, are risk factors for cardiovascular disease7, 8, 9, 10, 11. However, the precise biological role of BAT for TRL clearance remains unclear. Here we show that increased BAT activity induced by short-term cold exposure controls TRL metabolism in mice. Cold exposure drastically accelerated plasma clearance of triglycerides as a result of increased uptake into BAT, a pr! ocess crucially dependent on local LPL activity and transmembrane receptor CD36. In pathophysiological settings, cold exposure corrected hyperlipidemia and improved deleterious effects of insulin resistance. In conclusion, BAT activity controls vascular lipoprotein homeostasis by inducing a metabolic program that boosts TRL turnover and channels lipids into BAT. Activation of BAT might be a therapeutic approach to reduce elevated triglyceride concentrations and combat obesity in humans. View full text Figures at a glance * Figure 1: Cold exposure modulates fasting and postprandial triglyceride-rich lipoprotein levels. (,) Triglyceride () and cholesterol () FPLC profiles of pooled plasma from fasted FVB mice after 4 h and 24 h cold exposure at 4 °C. () Plasma triglycerides during an oral fat tolerance test in control and cold-exposed FVB mice. () FPLC lipoprotein profiling in control and cold-exposed FVB mice 2 h after an oral fat load. () Organ distribution of triolein-derived 3H-radioactivity in control and cold-exposed FVB mice 2 h after gavage. EpiWAT, epididymal white adipose tissue; SubWAT, subcutaneous white adipose tissue. Mean values ± s.e.m. with n = 12 in and n = 4 in . *P < 0.05; $P < 0.001. * Figure 2: Activated BAT is a central target organ for TRL uptake. (,) Plasma clearance of 59Fe-SPIO () and 3H-triolein-labeled TRLs () in control and cold-exposed C57BL/6 mice. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H-radioactivity () 15 min after intravenous injection. Mean values ± s.e.m. with n ≥ 5. () Representative transversal MRIs of a control and a cold-exposed wild-type FVB mouse before and approximately 10 min after injection of SPIO-labeled TRLs. Arrows in the top images point to BAT, whereas arrows in the bottom images indicate the liver. Scale bar, 1 cm. (,) Coronal MRIs of a representative cold mouse before and 10 min after () and 1 week after () injection of SPIO-TRLs with identical MRI settings. () Representative intravital confocal microscopy images of dissected BAT in a live cold-exposed FVB mouse 2 min (left) and 30 min (right) after QD-TRL (green) injection (arrows indicate QD-TRLs). FITC-dextran (red) to stain blood vessels and DAPI (blue) to label nuclei. Scale bar, 25 μm. () Representative tr! ansmission electron microscopy pictures of high-pressure frozen BAT samples from a SPIO-TRL–injected, cold-exposed FVB mouse. L, lipid droplet; M, mitochondrium; C, capillary. Top left scale bar, 5 μm; top middle scale bar, 1 μm; top right scale bar, 0.05 μm; bottom left scale bar, 1 μm; bottom middle scale bar, 0.05 μm; bottom right scale bar, 0.02 μm. *P < 0.05; &P < 0.01; $P < 0.001. * Figure 3: LPL and CD36 drive TRL clearance into BAT. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H () radioactivity 15 min after intravenous injection in cold FVB mice that were preinjected with tetrahydrolipstatin (THL) to inhibit LPL activity or with heparin to release LPL into circulation, respectively. Mean values ± s.e.m. with n ≥ 5. () Oral fat tolerance test in cold FVB mice pretreated with THL. Mean values ± s.e.m. with n = 5. () Relative mRNA expression in C57BL/6J BAT of several genes. The values for the lean control samples were arbitrarily set to 1 for each gene examined. AU, arbitrary units. () Determination of Cd36 and other fatty acid transporters mRNA copy numbers normalized to the housekeeping TATA-binding protein (Tbp) mRNA by TaqMan. () Consecutive FPLC analysis of TRL-3H-triolein and albumin-3H-oleate in cold-exposed wild-type and Cd36−/− littermates. (,) Organ distribution of 59Fe-SPIO () and triolein-derived 3H () radioactivity 15 min after intravenous injection of radiolabeled TR! Ls into Cd36−/− and wild-type littermates. Mean values ± s.e.m. with n ≥ 6. *P < 0.05; &P < 0.01; $P < 0.001. * Figure 4: BAT activation corrects hyperlipidemia and is not impaired in insulin resistance. (,) Triglyceride concentrations in hyperlipidemic Apoa5−/− mice during cold exposure () and photograph () of plasma after 24 h cold exposure. () Triglyceride and cholesterol FPLC profiling of pooled plasma from Apoa5−/− mice after 4 h and 24 h cold exposure. () Environmental scanning electron microscopy studies of brown adipose tissue from lean and obese control and cold-exposed mice. Scale bar, 25 μm. () Photographs of interscapular BAT in control and cold-exposed obese mice. Scale bar, 0.5 cm. (,) Combined oral glucose and fat tolerance test in lean and obese control and cold-exposed mice using 14C-deoxyglucose () and 3H-triolein () tracers. (,) Turnover kinetics () and organ uptake () of 3H-triolein-TRLs in lean and obese control and cold-exposed mice. Mean values ± s.e.m. with n = 6. *P < 0.05; &P < 0.01; $P < 0.001. NS, not significant. Author information * Author information * Supplementary information Affiliations * Institute of Biochemistry and Molecular Biology II: Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Alexander Bartelt, * Karoline Bruegelmann, * Barbara Freund, * Peter Nielsen, * Martin Merkel & * Joerg Heeren * Department of Electron Microscopy and Micro Technology, Heinrich-Pette Institute, Hamburg, Germany. * Oliver T Bruns, * Rudolph Reimer & * Heinz Hohenberg * Department of Diagnostic and Interventional Radiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Harald Ittrich, * Kersten Peldschus & * Michael G Kaul * Institute of Physical Chemistry, University of Hamburg, Hamburg, Germany. * Ulrich I Tromsdorf & * Horst Weller * Physical Chemistry, Technical University Dresden, Dresden, Germany. * Christian Waurisch & * Alexander Eychmüller * Department of Human Genetics, University of Leuven, Leuven, Belgium. * Philip L S M Gordts * III. Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. * Franz Rinninger * Asklepios Clinic St. Georg, Department of Internal Medicine, Hamburg, Germany. * Martin Merkel Contributions A.B. and J.H. designed the study, were involved in all aspects of the experiments and co-wrote the manuscript. O.T.B., R.R. and H.H. were responsible for electron microscopy and intravital imaging. H.I., K.P., O.T.B. and M.G.K. were responsible for MRI measurements. C.W., A.E., U.I.T., H.W., B.F. and P.N. were responsible for design and preparation of hydrophobic QD and SPIO, respectively. O.T.B., P.L.S.M.G., F.R., K.B., B.F., P.N. and M.M. were involved in turnover studies. All authors discussed the results and commented on the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding authors Correspondence to: * Alexander Bartelt or * Joerg Heeren Supplementary information * Author information * Supplementary information Movies * Supplementary Video 1 (23M) MRI of SPIO-TRL uptake into BAT.A representative MRI movie of a cold-exposed (cold) and a control mouse (control). After tail vein injection TRL labeled with super-paramagnetic iron-oxide nanocrystals (SPIO-TRL; start of the clock), liver contrast increases as a result of SPIO-TRL uptake in both mice. BAT contrast increase is only observed in the cold mouse. * Supplementary Video 2 (31M) Intravital imaging of BAT after QD-TRL injection. High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey). In cold-exposed mice, TRL which were labeled with hydrophobic fluorescent nanocrystals (QD-TRL; green) are injected via the tail vein. BAT-mediated processing of QD-TRL reveals a rapid attachment to the endothelium. Nuclei are stained with Hoechst (blue) and blood flow is visualized with FITC-dextran (red). * Supplementary Video 3 (45M) Intravital imaging of BODIPY-TRL uptake into BAT. High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey; movie is fourfold accelerated). In cold-exposed mice, TRL which were labeled with BODIPY-TG (BODIPY-TRL, red) are injected via the tail vein at the beginning of the movie. After approx. 2 min, 50 U heparin are injected and initially bound TRL are released from the vessel wall. * Supplementary Video 4 (30M) Intravital imaging of BODIPY-QD-double-labeled TRL uptake into BAT (heparin intervention). High-speed confocal intravital imaging was established to visualize the vascular circulation and structure of interscapular BAT in real time (reflection mode, grey). In cold-exposed mice, TRL which were double-labeled with BODIPY-TG and QD (BODIPY-TRL, red; QD-TRL, green) were injected via the tail vein 30 min before the movie starts. At the beginning of the movie only the QD signal is detectable. Next, 50 U heparin are injected, however, the QD signal cannot be released indicating internalization of TRL cores. Thereafter, a second bolus of double-labeled TRL is injected but the particles cannot bind to the endothelium and display prolonged circulation. * Supplementary Video 5 (20M) Intravital imaging of BODIPY-labeled TRL uptake into BAT (heparin intervention). This movie is identical to Supplementary Movie 4 except that only the BODIPY channel (BODIPY-TRL, red) is shown to demonstrate prolonged circulation of TRL while the binding to BAT endothelium is abolished in heparin-treated mice. * Supplementary Video 6 (15M) CD36-deficient mice after cold-exposure #1. A representative movie of a wild-type and Cd36-/- mouse. After 12 h cold exposure, Cd36-/- mice are characterized by low locomotor activity and noticeable shivering compared to wild-type control. * Supplementary Video 7 (26M) CD36-deficient mice after cold-exposure #2. Another example of a wild-type and Cd36-/- mouse. After 12 h cold exposure, Cd36-/- mice are characterized by low locomotor activity and noticeable shivering compared to wild-type control. * Supplementary Video 8 (15M) CD36-deficient mice after recovery. A representative movie of a wild-type and Cd36-/- mouse after 12 h recovery at room temperature. Under these conditions, Cd36-/- mice are indistinguishable from wild-type mice. PDF files * Supplementary Text and Figures (3M) Supplementary Figures and Tables Additional data - Hyperglycemia-induced cerebral hematoma expansion is mediated by plasma kallikrein
- Nat Med 17(2):206-210 (2011)
Nature Medicine | Letter Hyperglycemia-induced cerebral hematoma expansion is mediated by plasma kallikrein * Jia Liu1, 2, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Ben-Bo Gao1, 2, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Allen C Clermont1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Price Blair3 Search for this author in: * NPG journals * PubMed * Google Scholar * Tamie J Chilcote4 Search for this author in: * NPG journals * PubMed * Google Scholar * Sukanto Sinha4 Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Flaumenhaft3 Search for this author in: * NPG journals * PubMed * Google Scholar * Edward P Feener1, 2 Contact Edward P Feener Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:206–210Year published:(2011)DOI:doi:10.1038/nm.2295Received27 September 2010Accepted16 December 2010Published online23 January 2011 Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Hyperglycemia is associated with greater hematoma expansion and poor clinical outcomes after intracerebral hemorrhage. We show that cerebral hematoma expansion triggered by intracerebral infusion of autologous blood is greater in diabetic rats and mice compared to nondiabetic controls and that this augmented expansion is ameliorated by plasma kallikrein (PK) inhibition or deficiency. Intracerebral injection of purified PK augmented hematoma expansion in both diabetic and acutely hyperglycemic rats, whereas injection of bradykinin, plasmin or tissue plasminogen activator did not elicit such a response. This response, which occurs rapidly, was prevented by co-injection of the glycoprotein VI agonist convulxin and was mimicked by glycoprotein VI inhibition or deficiency, implicating an effect of PK on inhibiting platelet aggregation. We show that PK inhibits collagen-induced platelet aggregation by binding collagen, a response enhanced by elevated glucose concentrations. The ef! fect of hyperglycemia on hematoma expansion and PK-mediated inhibition of platelet aggregation could be mimicked by infusing mannitol. These findings suggest that hyperglycemia auguments cerebral hematoma expansion by PK-mediated osmotic-sensitive inhibition of hemostasis. View full text Figures at a glance * Figure 1: Effect of kallikrein-kinin inhibition on blood-induced hematoma expansion in brains of diabetic animals. (,) Representative images of the dorsal surface () and surface hematoma area as a percentage of hemisphere area () 48 h after intracerebral injection of autologous blood into the right hemisphere or PBS injection (sham) into the left hemisphere in nondiabetic (NDM; n = 15) and diabetic (DM; n = 11) rats. () Representative images of the dorsal surface and surface hematoma area as a percentage of hemisphere area in diabetic Akita mice (Ins2Akita, n = 10) and in nondiabetic littermate wild-type (WT) controls (n = 9) after PBS injection into the left hemisphere and autologous blood injection into the right hemisphere. () Surface hematoma area as a percentage of hemisphere area 48 h after intracerebral injection of autologous blood in nondiabetic rats systemically treated with vehicle (Veh, n = 4) or ASP-440 (440, n = 6) and in diabetic rats systemically treated with vehicle (n = 7), ASP-440 (n = 11), Hoe140 (Hoe, n = 8) or [des-Arg10]-Hoe140 (Des-Hoe, n = 5). () Surface hematoma! area as a percentage of hemisphere area 48 h after intracerebral injection of autologous blood mixed with PK-specific antibody into the right hemisphere or with control IgG into the left hemisphere of diabetic rats (n = 6). () Representative coronal slices 2 h after intracerebral injection of blood into the right hemisphere or PBS injection into the left hemisphere of diabetic Klkb1+/+ and Klkb1−/− mice. () Hemoglobin content of hemispheres subjected to autologous blood in Klkb1+/+ (n = 16), Klkb1+/− (n = 10) and Klkb1−/− (n = 23) mice. () Activated partial thromboplastin time (aPTT) in Klkb1+/+, Klkb1+/− and Klkb1−/− mice (n = 6 per group). *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars in ,,: 2 mm. Error bars represent mean ± s.e.m. * Figure 2: Effect of intracerebral injection of PK on hematoma expansion in brains of diabetic or acutely hyperglycemic rats. (,) Representative images of the dorsal surface and a coronal slice () and surface hematoma area as a percentage of hemisphere area () 48 h after intracerebral injection of PK into the right hemisphere or PBS injection (sham) into the left hemisphere in nondiabetic (NDM; n = 5) and diabetic (DM; n = 7) rats. Scale bar, 2 mm. (,) Hemoglobin () and carbonic anhydrase-1 (CA-1) levels () in a 5-mm coronal section of the hemisphere encompassing the injection site (n = 3–7 per group). () Time course of hematoma expansion after intracerebral injection of PK into the right hemisphere or PBS injection into the left hemisphere of diabetic rats (n = 7–8 per group). () Surface hematoma area as a percentage of hemisphere area in diabetic rats treated with insulin immediately before intracerebral PK injection (n = 8–12 per group). () Time course of blood glucose after intraperitoneal injection of saline (n = 11) or glucose (n = 15) in rats. () Hematoma area as a percentage of hemisp! here area 0.5 h after intracerebral PK or PBS injection in rats intraperitoneally injected with saline or glucose (n = 10–13 per group). () Surface hematoma area as a percentage of hemisphere area in nondiabetic and diabetic rats 0.5 h after intracerebral injection of PK or deactivated PK (De-PK) (n = 6–14 per group). *P < 0.05; **P < 0.01; ***P < 0.001. Error bars represent mean ± s.e.m. * Figure 3: Effect of PK on platelet aggregation and the effect of GPVI on cerebral hematoma expansion in animals. () Effect of PK on collagen-stimulated platelet aggregation. Data represent three independent experiments. () Effect of PK on thrombin- or ADP-induced platelet aggregation. Data represent three independent experiments. () Effect of glucose on PK-induced inhibition of collagen-stimulated platelet aggregation. Data represent three independent experiments. () Effects of PK (160 nM), prekallikrein (160 nM) and deactivated PK (De-PK, 160 nM) on collagen-induced platelet aggregation (n = 4 independent experiments). () Hemisphere hemoglobin content and () hematoma area as a percentage of hemisphere area in C57BL/6 wild-type (WT) mice subjected to JAQ1 or control rat IgG injection in the contralateral hemisphere (n = 6), C57BL/6 WT mice subjected to PBS injection (n = 10) and FcR γ-chain–deficient (Fcer1g−/−) mice subjected to PBS injection (n = 5). () Effect of convulxin on PK-induced hematoma expansion in diabetic rats (n = 6–19 rats per group). () Effects of PK (160 nM) ! and glucose (Glu, 25 mM) on CRP- or convulxin-induced platelet aggregation (n = 3–5 independent experiments). Platelet aggregation was measured with a platelet aggregometer in – and with a microplate reader in and . *P < 0.05; **P < 0.01; ***P < 0.001. Error bars represent mean ± s.e.m. * Figure 4: Effect of osmolarity on the binding of PK to collagen, collagen-stimulated platelet aggregation and hematoma expansion. (,) Sensorgram of the interaction of PK with immobilized collagen type I in the absence () or presence () of glucose. Data represent three independent experiments. () Effect of glucose on PK binding to aorta tissue (n = 5 pieces of rat aorta tissue). () Effects of hyperosmotic mannitol (Man) or hyperosmotic NaCl on PK-induced inhibition of collagen-stimulated platelet aggregation (n = 3 independent experiments). Glu, glucose. () Representative images of the dorsal surface and () surface hematoma area as a percentage of hemisphere area 2 h after intracerebral injection of PK or autologous blood into the right hemisphere and PBS injection (sham) into the left hemisphere of mannitol-treated nondiabetic rats (n = 6 per group). *P < 0.05; **P < 0.01. Scale bar, 2 mm. Error bars represent mean ± s.e.m. Author information * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Jia Liu & * Ben-Bo Gao Affiliations * Research Division, Joslin Diabetes Center, Boston, Massachusetts, USA. * Jia Liu, * Ben-Bo Gao, * Allen C Clermont & * Edward P Feener * Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA. * Jia Liu, * Ben-Bo Gao, * Allen C Clermont & * Edward P Feener * Department of Medicine, Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA. * Price Blair & * Robert Flaumenhaft * ActiveSite Pharmaceuticals, Inc., Berkeley, California, USA. * Tamie J Chilcote & * Sukanto Sinha Contributions J.L. initiated, designed and conducted most of the experiments and wrote the manuscript. B.-B.G. contributed to the design and performance of animal studies, biochemical analyses, platelet studies and manuscript writing. A.C.C. contributed to animal studies. P.B. and R.F. designed and performed studies using platelet aggregometer and contributed to data interpretation. T.J.C. and S.S. contributed to data interpretation and manuscript editing. E.P.F. designed and supervised the entire study and contributed to manuscript writing. Competing financial interests Tamie J. Chilcote and Sukanto Sinha are employees of and shareholders in ActiveSite Pharmaceuticals, which provided ASP-440. The Joslin Diabetes Center and ActiveSite Pharmaceuticals have submitted a patent application to the US Patent and Trademark Office and the European Patent Office on the methods for treatment of kallikrein-related disorders. Corresponding author Correspondence to: * Edward P Feener Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (274K) Supplementary Figures 1–8 and Supplementary Methods Additional data - The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44
- Nat Med 17(2):211-215 (2011)
Nature Medicine | Letter The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44 * Can Liu1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Kevin Kelnar3 Search for this author in: * NPG journals * PubMed * Google Scholar * Bigang Liu1 Search for this author in: * NPG journals * PubMed * Google Scholar * Xin Chen1, 2 Search for this author in: * NPG journals * PubMed * Google Scholar * Tammy Calhoun-Davis1 Search for this author in: * NPG journals * PubMed * Google Scholar * Hangwen Li1 Search for this author in: * NPG journals * PubMed * Google Scholar * Lubna Patrawala2 Search for this author in: * NPG journals * PubMed * Google Scholar * Hong Yan1 Search for this author in: * NPG journals * PubMed * Google Scholar * Collene Jeter1 Search for this author in: * NPG journals * PubMed * Google Scholar * Sofia Honorio1 Search for this author in: * NPG journals * PubMed * Google Scholar * Jason F Wiggins3 Search for this author in: * NPG journals * PubMed * Google Scholar * Andreas G Bader3 Search for this author in: * NPG journals * PubMed * Google Scholar * Randy Fagin4 Search for this author in: * NPG journals * PubMed * Google Scholar * David Brown3 Search for this author in: * NPG journals * PubMed * Google Scholar * Dean G Tang1, 2 Contact Dean G Tang Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:211–215Year published:(2011)DOI:doi:10.1038/nm.2284Received14 May 2010Accepted30 November 2010Published online16 January 2011 Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Cancer stem cells (CSCs), or tumor-initiating cells, are involved in tumor progression and metastasis1. MicroRNAs (miRNAs) regulate both normal stem cells and CSCs2, 3, 4, 5, and dysregulation of miRNAs has been implicated in tumorigenesis6. CSCs in many tumors—including cancers of the breast7, pancreas8, head and neck9, colon10, 11, small intestine12, liver13, stomach14, bladder15 and ovary16—have been identified using the adhesion molecule CD44, either individually or in combination with other marker(s). Prostate CSCs with enhanced clonogenic17 and tumor-initiating and metastatic18, 19 capacities are enriched in the CD44+ cell population, but whether miRNAs regulate CD44+ prostate cancer cells and prostate cancer metastasis remains unclear. Here we show, through expression analysis, that miR-34a, a p53 target20, 21, 22, 23, 24, was underexpressed in CD44+ prostate cancer cells purified from xenograft and primary tumors. Enforced expression of miR-34a in bulk or purifie! d CD44+ prostate cancer cells inhibited clonogenic expansion, tumor regeneration, and metastasis. In contrast, expression of miR-34a antagomirs in CD44− prostate cancer cells promoted tumor development and metastasis. Systemically delivered miR-34a inhibited prostate cancer metastasis and extended survival of tumor-bearing mice. We identified and validated CD44 as a direct and functional target of miR-34a and found that CD44 knockdown phenocopied miR-34a overexpression in inhibiting prostate cancer regeneration and metastasis. Our study shows that miR-34a is a key negative regulator of CD44+ prostate cancer cells and establishes a strong rationale for developing miR-34a as a novel therapeutic agent against prostate CSCs. View full text Figures at a glance * Figure 1: Underexpression and tumor-inhibitory effects of miR-34a. () Experimental scheme. (,) miR-34a levels in CD44+ xenograft (, PC4, LAPC4) or primary tumor (, HPCa) cells (mean percentage of marker-positive over marker-negative cells). (,) Orthotopic LAPC9 () and subcutaneous HPCa58 (; black, lenti-ctl; gray, lenti-34a; n = 10 and n = 7 for primary and secondary experiments, respectively) tumor growth (mean ± s.d.). () Lenti-34a mediated overexpression of miR-34a in purified CD44+ Du145 cells completely blocked subcutaneous tumor regeneration. () Subcutaneous tumor regeneration from purified CD44+ LAPC9 cells transfected with miR-NC or miR-34a. () Anti-miR-34a oligo transfection in purified CD44− Du145 cells promoted subcutaneous tumor growth. () Weight (mean ± s.d.) of LAPC9 tumors from bulk cells transfected with anti-NC or anti-34a oligos and implanted (100,000 cells) in the DP. Mice were killed at day 46. () Representative microphotographs showing increased lung metastasis by anti-34a (top, animal number and tumor weight; scale! bar, 100 μm; also see Supplementary Fig. 7c,d). * Figure 2: miR-34a inhibits prostate CSC properties. () Holoclone assays in Du145 cells. Cells transfected with miR-NC (NC) or miR-34a (34a) oligos were used in three experiments (Exp. I, 100 cells per well scored on day 9; Exp. II, 100 cells per well scored on day 13; Exp. III, 500 cells per well scored on day 7). () Clonogenic assays in Du145 cells. Cells (3,000 per well) were plated in Matrigel and colonies counted on day 13. NT, nontransfected. () Matrigel clonogenic assays in LAPC4 cells. Two experiments were performed (Exp. I, 1,250 cells per well scored on day 5, *P = 0.005; Exp. II, 25,000 cells per well scored on day 5, **P = 0.015). () Sphere assays in LAPC4 cells infected with lenti-ctl (C) or lenti-34a. Both primary and secondary spheres were scored on day 15. () Holoclone assays in PPC-1 cells (quantified on day 5). () Sphere assays in HPCa101 cells infected with lenti-ctl (C) or lenti-34a. Both primary and secondary spheres were scored 3 weeks later. (,) Sphere assays in purified CD44+ HPCa116 cells transfected w! ith NC or miR-34a oligos () or CD44− HPCa116 cells transfected with anti-NC or anti-34a oligos (). Spheres were scored on day 15. * Figure 3: Therapeutic effects of miR-34a. () Injections of miR-34a into the tail vein inhibited orthotopic PC3 tumor growth (n = 9 each). (–) Injections of miR-34a oligos into the tail vein inhibited metastasis (GFP+ foci in the endpoint lungs; mean ± s.d., n = 6 per group) of orthotopic LAPC9-GFP tumors () without significantly affecting tumor growth () and extended mouse survival (; Kaplan-Meier analysis and log-rank test). (,) The fourth set of therapeutic experiments in LAPC9 cells. Representative lung images (, animal number and tumor weight indicated on top; scale bar, 100 μm) and quantification of lung metastases (; mean ± s.d., n = 10 per group). * Figure 4: CD44 is a direct and functional target of miR-34a. () Representative CD44 immunohistochemistry images in Du145 tumors from cells infected with MSCV-PIG (control) or MSCV-34a vectors (western blot on the right) and PC3 tumors harvested from mice treated with miR-NC or miR-34a oligos. Scale bars, 10 μm. () miR-34a downregulates CD44 in Du145 (left) and PPC-1 (right) cells. Relative levels of CD44 indicated at the bottom. () Schematic of two putative miR-34a binding sites in the CD44 3′ UTR. () Luciferase experiments in Du145 cells (*P < 0.01). () Knockdown of CD44 inhibits LAPC4 tumor regeneration (Supplementary Fig. 12). (,) Knockdown of CD44 inhibits PC3 cell metastasis; shown are quantification () and images (; scale bar, 100 μm). (,) Invasion assays. miR-34a oligos inhibited Matrigel invasion of CD44+ Du145 cells (), and this inhibition was partially overcome by overexpression of a human CD44 cDNA lacking the miR-34a binding sites at the 3′ UTR (). Invasion expressed as values relative to the corresponding controls. ! () A schematic summary. The part highlighted in red refers to the novel findings of this study. Author information * Author information * Supplementary information Affiliations * Department of Molecular Carcinogenesis, the University of Texas M.D. Anderson Cancer Center, Science Park, Smithville, Texas, USA. * Can Liu, * Bigang Liu, * Xin Chen, * Tammy Calhoun-Davis, * Hangwen Li, * Hong Yan, * Collene Jeter, * Sofia Honorio & * Dean G Tang * Program in Molecular Carcinogenesis, The University of Texas Graduate School of Biomedical Sciences (GSBS), Houston, Texas, USA. * Can Liu, * Xin Chen, * Lubna Patrawala & * Dean G Tang * Mirna Therapeutics, Inc., Austin, Texas, USA. * Kevin Kelnar, * Jason F Wiggins, * Andreas G Bader & * David Brown * The Hospital at Westlake, Austin, Texas, USA. * Randy Fagin Contributions C.L., K.K., B.L., X.C. and L.P. designed and performed the experiments with help from C.J., T.C.-D., H.L., S.H., H.Y., J.F.W. and A.G.B., R.F. provided all HPCa samples. C.L. and D.G.T. prepared the manuscript. D.G.T., with help from D.B., designed the experiments and supervised the whole project. All authors discussed the results and commented on the manuscript. Competing financial interests K.K, J.F.W, A.G.B. and D.B are employees of Mirna Therapeutics, Inc., which develops miRNA-based therapeutics. Corresponding author Correspondence to: * Dean G Tang Supplementary information * Author information * Supplementary information PDF files * Supplementary Text and Figures (2M) Supplementary Results, Supplementary Methods, Supplementary Figures 1–15 and Supplementary Tables 1 and 2 Additional data - The Ngal reporter mouse detects the response of the kidney to injury in real time
- Nat Med 17(2):216-222 (2011)
Nature Medicine | Technical Report The Ngal reporter mouse detects the response of the kidney to injury in real time * Neal Paragas1, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Andong Qiu1, 5 Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyin Zhang1 Search for this author in: * NPG journals * PubMed * Google Scholar * Benjamin Samstein1 Search for this author in: * NPG journals * PubMed * Google Scholar * Shi-Xian Deng1 Search for this author in: * NPG journals * PubMed * Google Scholar * Kai M Schmidt-Ott2 Search for this author in: * NPG journals * PubMed * Google Scholar * Melanie Viltard1 Search for this author in: * NPG journals * PubMed * Google Scholar * Wenqiang Yu1 Search for this author in: * NPG journals * PubMed * Google Scholar * Catherine S Forster1 Search for this author in: * NPG journals * PubMed * Google Scholar * Gangli Gong1 Search for this author in: * NPG journals * PubMed * Google Scholar * Yidong Liu1 Search for this author in: * NPG journals * PubMed * Google Scholar * Ritwij Kulkarni1 Search for this author in: * NPG journals * PubMed * Google Scholar * Kiyoshi Mori3 Search for this author in: * NPG journals * PubMed * Google Scholar * Avtandil Kalandadze1 Search for this author in: * NPG journals * PubMed * Google Scholar * Adam J Ratner1 Search for this author in: * NPG journals * PubMed * Google Scholar * Prasad Devarajan4 Search for this author in: * NPG journals * PubMed * Google Scholar * Donald W Landry1 Search for this author in: * NPG journals * PubMed * Google Scholar * Vivette D'Agati1 Search for this author in: * NPG journals * PubMed * Google Scholar * Chyuan-Sheng Lin1 Search for this author in: * NPG journals * PubMed * Google Scholar * Jonathan Barasch1 Contact Jonathan Barasch Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:216–222Year published:(2011)DOI:doi:10.1038/nm.2290Received09 December 2009Accepted12 September 2010Published online16 January 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Many proteins have been proposed to act as surrogate markers of organ damage, yet for many candidates the essential biomarker characteristics that link the protein to the injured organ have not yet been described. We generated an Ngal reporter mouse by inserting a double-fusion reporter gene encoding luciferase-2 and mCherry (Luc2-mC) into the Ngal (Lcn2) locus. The Ngal-Luc2-mC reporter accurately recapitulated the endogenous message and illuminated injuries in vivo in real time. In the kidney, Ngal-Luc2-mC imaging showed a sensitive, rapid, dose-dependent, reversible, and organ- and cell-specific relationship with tubular stress, which correlated with the level of urinary Ngal (uNgal). Unexpectedly, specific cells of the distal nephron were the source of uNgal. Cells isolated from Ngal-Luc2-mC mice also revealed both the onset and the resolution of the injury, and the actions of NF-κB inhibitors and antibiotics during infection. Thus, imaging of Ngal-Luc2-mC mice and cell! s identified injurious and reparative agents that affect kidney damage. View full text Figures at a glance * Figure 1: Ngal-Luc2-mC visualized kidney damage in real time in vivo. () Bioluminescent radiation from heterozygous Ngal-Luc2-mC female mice subjected to left kidney ischemia-reperfusion (I/R) for 15 or 30 min. Time after I/R is indicated at top. () Ngal-Luc2-mC radiation 12 h after injury, imaged from injured kidney and contralateral, uninjured kidney. () Fold change in bioluminescence from a constant region of interest in the ischemic mice in . Kid, kidney. () Immunoblot of Ngal in the urine from the ischemic mouse (15 min) shown in . Recombinant mouse nonglycosylated Ngal was used as a standard. () Biouminescence from heterozygous Ngal-Luc2-mC albino female mice subjected to bilateral ischemia for 15 min. () sCr concentration, fold change in Ngal-Luc2 activity and fold change in uCr concentration after 15 min ischemia; lines show average results, horizontal and vertical bars show median ± s.e.m. from four (Ngal-Luc2-mC), ten (sCr) or ten (uCr) experiments. () Immunoblot showing uNgal protein in the urine of the mouse in . * Figure 2: Ngal-Luc2-mC reported kidney cellular damage in vivo induced by cisplatin and lipid A. () Ngal-Luc2-mC expression in both kidneys of a mouse 0 h and 168 h after cisplatin (20 mg kg−1) exposure. () Close-up view of kidneys and immunoblot showing uNgal from mouse in at 168 h. () Fluorescence from Ngal-Luc2-mC kidney cells treated with cisplatin (10 μM). () Ngal-Luc2-mC fluorescence elicited by lipid A treatment, with time and dosage indicated at top. Low-level expression of Ngal-Luc2-mC was also seen in the skin of the feet, similar to expression of TLR4 (ref. 40). () Average Ngal-Luc2-mC fluorescence (as in ) and sCr levels 24 h after exposure to lipid A (n = 3). () In situ hybridization showing Ngal mRNA in TAL and collecting ducts in the outer stripe of the inner medulla. () H&E staining showing cast formation 24 h after a 5-mg-kg−1 lipid A challenge. In ,, arrowheads indicate presumptive intercalated cells where Ngal RNA was localized; asterisks mark casts and cellular debris in collecting ducts. High-powered images of boxed regions are shown at the bot! tom. Scale bar, 10 μm. * Figure 3: Damaged nephron is the source of kidney Ngal. () In situ hybridization showing Ngal mRNA expression (purple) in dissected reporter kidney. Scale bar, 1 mm. (–) Paraffin sections of kidney. Top images, Ngal mRNA hybridization; bottom images, hematoxylin and eosin staining. Ngal was expressed in the outer stripe of the outer medulla and cortical TAL (in medullary rays) containing casts (asterisks, ), in collecting ducts (open arrowheads, ) containing casts (asterisks, ) and in the macula densa of the distal tube (Dt, open arrowheads, ), which had undergone epithelial flattening and cast formation (asterisks, ), but not in the necrotic pars recta of proximal tubules (Pt; filled arrowheads, or in glomeruli (G; )). Scale bars, 100 μm. () High-magnification mRNA hybridization image showing Ngal specifically expressed by intercalated cells (open arrowheads). Filled arrowhead, necrotic pars recta. Scale bars in –, 10 μm. () Anti–v-ATPase immunohistochemistry costained with Ngal mRNA in collecting-duct cells. v-ATPase ma! rks the apical surface of α-type intercalated cells (arrowheads), showing that Ngal is expressed from these cells. () Ngal mRNA and v-ATPase costaining as in , in outer medullary collecting ducts. Open arrowheads, α-type intercalated cells expressing Ngal; filled arrowheads, adjacent β-type intercalated cells with no Ngal. (,) Ngal mRNA expression (in situ hybridization, ) and H&E staining of the ischemic zone () in a polar (segmental) renal artery ischemia. The tissue was collected 24 h after a 30-min ischemia treatment. Closed arrowheads indicate the boundary of damaged and undamaged tubules in the papilla, and open arrowheads mark the width of the papilla. Scale bars in and , 300 μm. * Figure 4: Volume depletion did not activate Ngal reporter expression. (,) Effects of volume depletion on serum sodium () and sCr levels (). Graphs show mean and s.e.m. (n = 3). () Ngal-Luc2 fluorescence in mice after simple volume depletion. Kidney and testis are circled; testis serves as an internal positive control, as male mice tonically express Ngal in testis. * Figure 5: Expression of Ngal reflects pharmacological interventions. () Ngal-Luc2 expression in primary kidney cells (105 cells per well) isolated from reporter mice treated with uropathogenic E. coli (CFT073; 104 CFU ml−1) and/or gentamicin (100 μg ml−1; pretreated, −1 h, or post-treated, 1 and 6 h). Graph shows mean and s.e.m. (n = 3, independent experiments). () Ngal-Luc2 expression as in , induced by lipid A (4 μg ml−1) plus pretreatment with either a known NF-κB inhibitor, MG132 (0.5–5 μM), novel NF-κB inhibitors (inhib A–D; 5 μM) or DMSO control. Graph shows mean and s.e.m. (n = 3 independent experiments). () Expression of distal tubular markers Aqp2 and Umod and proximal tubular marker Aqp1 in primary cells isolated from the inner medulla and papilla of Ngal-Luc2-mC kidneys, relative to expression in cortical cells (n = 6). Ratio of gene expression in medullary cells/expression in cortical cells is plotted; error bars show s.e.m. () Ngal-Luc2 expression in inner medullary and papillary primary cells (medullary cells) ! and cortical cells, untreated (control) or treated with lipid A (4 μg ml−1, 24 h). Graphs show mean and s.e.m.; inset, net change in fluorescence from the control. Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Neal Paragas & * Andong Qiu Affiliations * College of Physicians and Surgeons of Columbia University, New York, New York, USA. * Neal Paragas, * Andong Qiu, * Qingyin Zhang, * Benjamin Samstein, * Shi-Xian Deng, * Melanie Viltard, * Wenqiang Yu, * Catherine S Forster, * Gangli Gong, * Yidong Liu, * Ritwij Kulkarni, * Avtandil Kalandadze, * Adam J Ratner, * Donald W Landry, * Vivette D'Agati, * Chyuan-Sheng Lin & * Jonathan Barasch * Max-Delbruck Center for Molecular Medicine Berlin-Buch, Berlin, Germany. * Kai M Schmidt-Ott * Kyoto University Graduate School of Medicine, Kyoto, Japan. * Kiyoshi Mori * Cincinnati Children's Hospital, Cincinnati, Ohio, USA. * Prasad Devarajan Contributions N.P., A.Q. and C.-S.L. created the Ngal reporter mouse; Q.Z. and B.S. performed surgeries; S.-X.D., G.G., Y.L., D.W.L. created NF-κB inhibitors; N.P., R.K. and A.J.R. studied bacteria-induced Ngal expression; V.D. evaluated the pattern of Ngal expression; K.M.S.-O., M.V., W.Y., C.S.F., K.M., A.K. and P.D. analyzed data; N.P., A.Q. and J.B. and wrote the paper; A.Q., N.P. and J.B. invented the luminescent mouse. Competing financial interests Columbia University and Cincinnati Children's Hospital Medical Center have received licensing fees from Biosite (Inverness Medical) and Abbott Diagnostics. Corresponding author Correspondence to: * Jonathan Barasch Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (2M) Supplementary Methods, Supplementary Table 1 and Supplementary Figures 1–12 Additional data - Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy
- Nat Med 17(2):223-228 (2011)
Nature Medicine | Technical Report Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy * Robert P J Barretto1, 6 Search for this author in: * NPG journals * PubMed * Google Scholar * Tony H Ko1, 6 Search for this author in: * NPG journals * PubMed * Google Scholar * Juergen C Jung1, 6 Search for this author in: * NPG journals * PubMed * Google Scholar * Tammy J Wang1 Search for this author in: * NPG journals * PubMed * Google Scholar * George Capps1 Search for this author in: * NPG journals * PubMed * Google Scholar * Allison C Waters1 Search for this author in: * NPG journals * PubMed * Google Scholar * Yaniv Ziv1 Search for this author in: * NPG journals * PubMed * Google Scholar * Alessio Attardo1 Search for this author in: * NPG journals * PubMed * Google Scholar * Lawrence Recht2, 3 Search for this author in: * NPG journals * PubMed * Google Scholar * Mark J Schnitzer1, 4, 5 Contact Mark J Schnitzer Search for this author in: * NPG journals * PubMed * Google Scholar * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:223–228Year published:(2011)DOI:doi:10.1038/nm.2292Received05 November 2010Accepted07 October 2010Published online16 January 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The combination of intravital microscopy and animal models of disease has propelled studies of disease mechanisms and treatments. However, many disorders afflict tissues inaccessible to light microscopy in live subjects. Here we introduce cellular-level time-lapse imaging deep within the live mammalian brain by one- and two-photon fluorescence microendoscopy over multiple weeks. Bilateral imaging sites allowed longitudinal comparisons within individual subjects, including of normal and diseased tissues. Using this approach, we tracked CA1 hippocampal pyramidal neuron dendrites in adult mice, revealing these dendrites' extreme stability and rare examples of their structural alterations. To illustrate disease studies, we tracked deep lying gliomas by observing tumor growth, visualizing three-dimensional vasculature structure and determining microcirculatory speeds. Average erythrocyte speeds in gliomas declined markedly as the disease advanced, notwithstanding significant incr! eases in capillary diameters. Time-lapse microendoscopy will be applicable to studies of numerous disorders, including neurovascular, neurological, cancerous and trauma-induced conditions. View full text Figures at a glance * Figure 1: Chronic mouse preparation for repeated imaging of deep brain tissues using microendoscopy. () The experiment begins with implantation of imaging guide tubes, one into each hemisphere of the mouse′s brain. After the mouse recovers from surgery, micro-optical probes can be repeatedly inserted into the guide tubes to allow time-lapse imaging. () Three microendoscope probes, a 500-μm-diameter singlet, a 1-mm-diameter singlet and a 1-mm-diameter compound doublet. Scale bar, 1 mm. () Schematic of a microscope objective lens coupling illumination into a 500-μm-diameter singlet microendoscope probe, such as that shown in . * Figure 2: Time-lapse two-photon microendoscopy of CA1 hippocampal neurons. Time-lapse image sequences of CA1 pyramidal neurons in three Thy1-GFP mice. (,) Two-dimensional projections of three-dimensional stacks containing four image slices acquired at 4.2-μm axial spacing over 16.8 μm in depth. shows enlargements of the boxed area in . () Two-dimensional projections of three-dimensional stacks acquired at 3-μm axial spacing over approximately 540 μm in depth. () Enlarged, single-image frames revealing spiny dendrites. Scale bars in , , and are 100, 25, 50 and 5 μm, respectively. * Figure 3: Time-lapse microendoscopy of CA1 microvasculature shows normal blood vessel morphologies are stable over time. () Two-dimensional projection of a three-dimensional stack of 220 images of dye-labeled vasculature acquired at ~3-μm increments over ~660 μm in depth. Supplementary Video 1 shows the entire image stack. () Time-lapse image sequence acquired by one-photon microendoscopy. () Time-lapse sequence of two-photon image stacks, each composed of 40–50 images acquired approximately 3.7 μm apart in depth and projected to two dimensions. Figure 4b shows dye-labeled tumor vessels from the opposing (experimental) hippocampus in the same mouse. (,) One-photon image of CA1 blood vessels () and corresponding microcirculatory speed map (), determined by high-speed (100-Hz) imaging and cross-correlation analysis. Supplementary Video 2 shows blood flow from this same field of view. Scale bars are 100 μm in – and 50 μm in . * Figure 4: Time-lapse imaging of glioma angiogenesis in mouse CA1 reveals progressive distortions to vascular geometry and reduced microcirculatory speeds. () Dual-color image of GFP-expressing mouse glioma cells (green) and rhodamine-dextran–labeled microvasculature (red), acquired in a live mouse by one-photon fluorescence microendoscopy on day 3 after glioma cell inoculation. (,) Time-lapse sequences of two-photon microendoscopy image stacks, projected to two dimensions, showing the progressive distortion of the microvasculature due to glioma angiogenesis. Each stack contained 40–50 images acquired 3.7 μm apart in depth. Figure 3c shows images of the opposing (control) hippocampus from the same mouse used in . () Maps of average erythrocyte speed, determined from 10-s videos acquired at 100 Hz by one-photon microendoscopy, in left (control) and right (experimental) hemispheres of a live mouse on day 20 after glioma inoculation. Scale bars, 100 μm. * Figure 5: Quantitative tracking of glioma angiogenesis in CA1 hippocampus shows tumor vessels broaden in diameter but undergo marked declines in flow speed. Vessel diameters and erythrocyte flow speeds were monitored as a function of elapsed time after an initial surgery and glioma cell implantation on day 0. Not all mice were imaged at identical time points, so time values were binned into 3-d intervals. () Plots of flow speed versus vessel diameter, for control tissue (left) and tumor sites (right), in which each data point represents an individual vessel segment. Unlike at control sites, at tumor sites the data reveal an overall progressive decrease in flow speeds and increase in vessel diameters as the disease advanced (n = 10 mice). () Population averages of erythrocyte flow speed (left) and vessel diameter (right) are plotted (mean ± s.e.m.) versus elapsed time. Mean tumor vessel diameters and flow speeds differed significantly from control values (P < 1 × 10−6 and P < 1 × 10−15, respectively; Mann-Whitney U test). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Robert P J Barretto, * Tony H Ko & * Juergen C Jung Affiliations * James H. Clark Center for Biomedical Engineering & Sciences, Stanford University, Stanford, California, USA. * Robert P J Barretto, * Tony H Ko, * Juergen C Jung, * Tammy J Wang, * George Capps, * Allison C Waters, * Yaniv Ziv, * Alessio Attardo & * Mark J Schnitzer * Department of Neurology and Neurological Sciences, Stanford University, Stanford, California, USA. * Lawrence Recht * Department of Neurosurgery, Stanford University, Stanford, California, USA. * Lawrence Recht * Howard Hughes Medical Institute, Stanford University, Stanford, California, USA. * Mark J Schnitzer * CNC Program, Stanford University, Stanford, California, USA. * Mark J Schnitzer Contributions R.P.J.B. designed experiments, developed tracking of neuronal dendrites, performed the study on CA1 neuron stability, analyzed the neuronal histology data, validated the algorithm for computing erythrocyte speeds and computed relationships between vessel diameters and speeds. T.H.K. designed experiments, performed the glioma experiments and computed flow speeds and vessel sizes. J.C.J. designed experiments, developed the chronic preparation and tested it for imaging neurons and gliomas. T.J.W. and G.C. performed neuronal imaging and contributed to the glioma experiments. A.C.W. developed bilateral imaging, performed neuronal imaging and contributed to the glioma experiments. Y.Z. developed and performed striatal imaging. A.A. performed histological analyses and analyzed vessel branching ratios. L.R. designed experiments and supervised the glioma study. M.J.S. designed experiments, performed statistical testing, initiated and supervised the project and wrote the paper. All au! thors edited the paper. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * Mark J Schnitzer Supplementary information * Abstract * Author information * Supplementary information Movies * Supplementary Video 1 (3M) Three-dimensional image stack of hippocampal blood vessels acquired in a live mouse by two-photon microendoscopy and intravascular injection of fluorescein-dextran. * Supplementary Video 2 (8M) Hippocampal microcirculation in normal tissue imaged by high-speed one-photon microendoscopy and intravascular injection of fluorescein-dextran. * Supplementary Video 3 (6M) High-speed imaging of microcirculation in a hippocampal glioma using one-photon microendoscopy. PDF files * Supplementary Text and Figures (1M) Supplementary Figures 1–4 and Supplementary Methods Additional data - Inhibition of aldehyde dehydrogenase-2 suppresses cocaine seeking by generating THP, a cocaine use–dependent inhibitor of dopamine synthesis
- Nat Med 17(2):229 (2011)
Nature Medicine | Addendum Inhibition of aldehyde dehydrogenase-2 suppresses cocaine seeking by generating THP, a cocaine use–dependent inhibitor of dopamine synthesis * Lina Yao Search for this author in: * NPG journals * PubMed * Google Scholar * Peidong Fan Search for this author in: * NPG journals * PubMed * Google Scholar * Maria Arolfo Search for this author in: * NPG journals * PubMed * Google Scholar * Zhang Jiang Search for this author in: * NPG journals * PubMed * Google Scholar * M Foster Olive Search for this author in: * NPG journals * PubMed * Google Scholar * Jeff Zablocki Search for this author in: * NPG journals * PubMed * Google Scholar * Hai-Ling Sun Search for this author in: * NPG journals * PubMed * Google Scholar * Nancy Chu Search for this author in: * NPG journals * PubMed * Google Scholar * Jeongrim Lee Search for this author in: * NPG journals * PubMed * Google Scholar * Hee-Yong Kim Search for this author in: * NPG journals * PubMed * Google Scholar * Kwan Leung Search for this author in: * NPG journals * PubMed * Google Scholar * John Shryock Search for this author in: * NPG journals * PubMed * Google Scholar * Brent Blackburn Search for this author in: * NPG journals * PubMed * Google Scholar * Ivan Diamond Search for this author in: * NPG journals * PubMed * Google ScholarJournal name:Nature MedicineVolume: 17,Page:229Year published:(2011)DOI:doi:10.1038/nm0211-229Published online04 February 2011 Article tools * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg Nat. Med.16, 1024–1028 (2010); published online 22 August 2010; corrected online 26 August 2010; addendum published after print 13 January 2011. Nature Medicine has become aware that CVT-10216, the selective ALDH-2 inhibitor originally reported in this study, is not available from Gilead Sciences, the institution to which the corresponding author of the paper is affiliated. We wish to alert our readers of this situation, as it contravenes our editorial policy on material sharing (http://www.nature.com/authors/editorial_policies/availability.html). Additional data
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