Large intergenic non-coding RNA-RoR modulates reprogramming of human induced pluripotent stem cells

Abstract: The conversion of lineage-committed cells to induced pluripotent stem cells (iPSCs) by reprogramming is accompanied by a global remodeling of the epigenome, resulting in altered patterns of gene expression.

Abstract: Here we characterize the transcriptional reorganization of large intergenic non-coding RNAs (lincRNAs) that occurs upon derivation of human iPSCs and identify numerous lincRNAs whose expression is linked to pluripotency.

Abstract: Among these, we defined ten lincRNAs whose expression was elevated in iPSCs compared with embryonic stem cells, suggesting that their activation may promote the emergence of iPSCs.

Abstract: Supporting this, our results indicate that these lincRNAs are direct targets of key pluripotency transcription factors.

Abstract: Using loss-of-function and gain-of-function approaches, we found that one such lincRNA (lincRNA-RoR) modulates reprogramming, thus providing a first demonstration for critical functions of lincRNAs in the derivation of pluripotent stem cells.


Extracellular vesicle isolation and characterization: toward clinical application.

Abstract: Two broad categories of extracellular vesicles (EVs), exosomes and shed microvesicles (sMVs), which differ in size distribution as well as protein and RNA profiles, have been described.

Abstract: EVs are known to play key roles in cell-cell communication, acting proximally as well as systemically.

Abstract: This Review discusses the nature of EV subtypes, strategies for isolating EVs from both cell-culture media and body fluids, and procedures for quantifying EVs.

Abstract: We also discuss proteins selectively enriched in exosomes and sMVs that have the potential for use as markers to discriminate between EV subtypes, as well as various applications of EVs in clinical diagnosis.


Up-regulated NRIP2 in colorectal cancer initiating cells modulates the Wnt pathway by targeting RORβ

Abstract: BACKGROUND Colorectal cancer remains one of the most common malignant tumors worldwide.

Abstract: Colorectal cancer initiating cells (CCICs) are a small subpopulation responsible for malignant behaviors of colorectal cancer.

Abstract: Aberrant activation of the Wnt pathways regulates the self-renewal of CCIC.

Abstract: However, the underlying mechanism(s) remain poorly understood.

Abstract: METHODS Via retroviral library screening, we identified Nuclear Receptor-Interacting Protein 2 (NRIP2) as a novel interactor of the Wnt pathway from enriched colorectal cancer colosphere cells.

Abstract: The expression levels of NRIP2 and retinoic acid-related orphan receptor β (RORβ) were further examined by FISH, qRT-PCR, IHC and Western blot.

Abstract: NRIP2 overexpressed and knockdown colorectal cancer cells were produced to study the role of NRIP2 in Wnt pathway.

Abstract: We also verified the binding between NRIP2 and RORβ and investigated the effect of RORβ on CCICs both in vitro and in vivo.

Abstract: Genechip-scanning speculated downstream target HBP1.

Abstract: Western blot, ChIP and luciferase reporter were carried to investigate the interaction between NRIP2, RORβ, and HBP1.

Abstract: RESULTS NRIP2 was significantly up-regulated in CCICs from both cell lines and primary colorectal cancer tissues.

Abstract: Reinforced expression of NRIP2 increased Wnt activity, while silencing of NRIP2 attenuated Wnt activity.

Abstract: The transcription factor RORβ was a key target through which NRIP2 regulated Wnt pathway activity.

Abstract: RORβ was a transcriptional enhancer of inhibitor HBP1 of the Wnt pathway.

Abstract: NRIP2 prevented RORβ to bind with downstream HBP1 promoter regions and reduced the transcription of HBP1.

Abstract: This, in turn, attenuated the HBP1-dependent inhibition of TCF4-mediated transcription.

Abstract: CONCLUSIONS NRIP2 is a novel interactor of the Wnt pathway in colorectal cancer initiating cells.

Abstract: interactions between NRIP2, RORβ, and HBP1 mediate a new mechanism for CCIC self-renewal via the Wnt activity.


Speed Limits for Nonvesicular Intracellular Sterol Transport.

Abstract: Sterol transport between the endoplasmic reticulum (ER) and plasma membrane (PM) occurs by nonvesicular mechanisms requiring sterol transport proteins (STPs).

Abstract: Here we examine the idea that transport is enhanced at membrane contact sites where the ER is closely apposed to the PM.

Abstract: We conclude that sterol desorption from the membrane, rather than STP-mediated diffusion, is rate limiting in the cellular context, so there is no apparent kinetic benefit to having STP-mediated sterol transfer occur at contact sites.

Abstract: Contact sites may instead compartmentalize lipid synthesis or transport machinery, providing opportunities for regulation.


Misunderstandings in prescribing decisions in general practice: qualitative study.

Abstract: OBJECTIVES To identify and describe misunderstandings between patients and doctors associated with prescribing decisions in general practice.

Abstract: DESIGN Qualitative study.

Abstract: SETTING 20 general practices in the West Midlands and south east England.

Abstract: PARTICIPANTS 20 general practitioners and 35 consulting patients.

Abstract: MAIN OUTCOME MEASURES Misunderstandings between patients and doctors that have potential or actual adverse consequences for taking medicine.

Abstract: RESULTS 14 categories of misunderstanding were identified relating to patient information unknown to the doctor, doctor information unknown to the patient, conflicting information, disagreement about attribution of side effects, failure of communication about doctor's decision, and relationship factors.

Abstract: All the misunderstandings were associated with lack of patients' participation in the consultation in terms of the voicing of expectations and preferences or the voicing of responses to doctors' decisions and actions.

Abstract: They were all associated with potential or actual adverse outcomes such as non-adherence to treatment.

Abstract: Many were based on inaccurate guesses and assumptions.

Abstract: In particular doctors seemed unaware of the relevance of patients' ideas about medicines for successful prescribing.

Abstract: CONCLUSIONS Patients' participation in the consultation and the adverse consequences of lack of participation are important.

Abstract: The authors are developing an educational intervention that builds on these findings.


Phase-plate cryo-EM structure of a biased agonist-bound human GLP-1 receptor–Gs complex

Abstract: The class B glucagon-like peptide-1 (GLP-1) G protein-coupled receptor is a major target for the treatment of type 2 diabetes and obesity.

Abstract: Endogenous and mimetic GLP-1 peptides exhibit biased agonism—a difference in functional selectivity—that may provide improved therapeutic outcomes.

Abstract: Here we describe the structure of the human GLP-1 receptor in complex with the G protein-biased peptide exendin-P5 and a Gαs heterotrimer, determined at a global resolution of 3.3 Å. At the extracellular surface, the organization of extracellular loop 3 and proximal transmembrane segments differs between our exendin-P5-bound structure and previous GLP-1-bound GLP-1 receptor structure.

Abstract: At the intracellular face, there was a six-degree difference in the angle of the Gαs–α5 helix engagement between structures, which was propagated across the G protein heterotrimer.

Abstract: In addition, the structures differed in the rate and extent of conformational reorganization of the Gαs protein.

Abstract: Our structure provides insights into the molecular basis of biased agonism.


Therapeutic equivalence of alendronate 70 mg once-weekly and alendronate 10 mg daily in the treatment of osteoporosis. Alendronate Once-Weekly Study Group.

Abstract: Dosing convenience is a key element in the effective management of any chronic disease, and is particularly important in the long-term management of osteoporosis.

Abstract: Less frequent dosing with any medication may enhance compliance, thereby maximizing the effectiveness of therapy.

Abstract: Animal data support the rationale that once-weekly dosing with alendronate 70 mg (7 times the daily oral treatment dose) could provide similar efficacy to daily dosing with alendronate 10 mg due to its long duration of effect in bone.

Abstract: In addition, dog studies suggest that the potential for esophageal irritation, observed with daily oral bisphosphonates, may be substantially reduced with once-weekly dosing.

Abstract: This dosing regimen would provide patients with increased convenience and would be likely to enhance patient compliance.

Abstract: We compared the efficacy and safety of treatment with oral once-weekly alendronate 70 mg (N=519), twice-weekly alendronate 35 mg (N=369), and daily alendronate 10 mg (N=370) in a one-year, double-blind, multicenter study of postmenopausal women (ages 42 to 95) with osteoporosis (bone mineral density [BMD] of either lumbar spine or femoral neck at least 2.5 SDs below peak premenopausal mean, or prior vertebral or hip fracture).

Abstract: The primary efficacy endpoint was the comparability of increases in lumbar spine BMD, using strict pre-defined equivalence criteria.

Abstract: Secondary endpoints included changes in BMD at the hip and total body and rate of bone turnover, as assessed by biochemical markers.

Abstract: Both of the new regimens fully satisfied the equivalence criteria relative to daily therapy.

Abstract: Mean increases in lumbar spine BMD at 12 months were: 5.1% (95% CI 4.8, 5.4) in the 70 mg once-weekly group, 5.2% (4.9, 5.6) in the 35 mg twice-weekly group, and 5.4% (5.0, 5.8) in the 10 mg daily treatment group.

Abstract: Increases in BMD at the total hip, femoral neck, trochanter, and total body were similar for the three dosing regimens.

Abstract: All three treatment groups similarly reduced biochemical markers of bone resorption (urinary N-telopeptides of type I collagen) and bone formation (serum bone-specific alkaline phosphatase) into the middle of the premenopausal reference range.

Abstract: All treatment regimens were well tolerated with a similar incidence of upper GI adverse experiences.

Abstract: There were fewer serious upper GI adverse experiences and a trend toward a lower incidence of esophageal events in the once-weekly dosing group compared to the daily dosing group.

Abstract: These data are consistent with preclinical animal models, and suggest that once-weekly dosing has the potential for improved upper GI tolerability.

Abstract: Clinical fractures, captured as adverse experiences, were similar among the groups.

Abstract: We conclude that the alendronate 70 mg once-weekly dosing regimen will provide patients with a more convenient, therapeutically equivalent alternative to daily dosing, and may enhance compliance and long-term persistence with therapy.


Association of Toll-Like Receptor 4 on Human Monocyte Subsets and Vulnerability Characteristics of Coronary Plaque as Assessed by 64-Slice Multidetector Computed Tomography.

Abstract: BACKGROUND Although Toll-like receptor 4 (TLR-4) is involved in monocyte activation in patients with accelerated forms of atherosclerosis, the relationship between the expression of TLR-4 on circulating monocytes and coronary plaque vulnerability has not previously been evaluated.

Abstract: We investigated this relationship using 64-slice multidetector computed tomography (MDCT) in patients with stable angina pectoris (SAP).Methods and Results:We enrolled 65 patients with SAP who underwent MDCT.

Abstract: Three monocyte subsets (CD14++CD16-, CD14++CD16+, and CD14+CD16+) and expression of TLR-4 were measured by flow cytometry.

Abstract: Intracoronary plaques were assessed by 64-slice MDCT.

Abstract: We defined vulnerability of intracoronary plaques according to the presence of positive remodeling (remodeling index >1.05) and/or low CT attenuation (<35 HU).

Abstract: The circulating CD14++CD16+monocytes more frequently expressed TLR-4 than CD14++CD16-and CD14+CD16+monocytes (P<0.001).

Abstract: The relative proportion of the expression of TLR-4 on CD14++CD16+monocytes was significantly greater in patients with vulnerable plaque compared with those without (10.4 [4.1-14.5] % vs. 4.5 [2.8-7.8] %, P=0.012).

Abstract: In addition, the relative proportion of TLR-4 expression on CD14++CD16+monocytes positively correlated with the remodeling index (r=0.28, P=0.025) and negatively correlated with CT attenuation value (r=-0.31, P=0.013).

Abstract: CONCLUSIONS Upregulation of TLR-4 on CD14++CD16+monocytes might be associated with coronary plaque vulnerability in patients with SAP.


Expandable Cardiovascular Progenitor Cells Reprogrammed from Fibroblasts.

Abstract: Stem cell-based approaches to cardiac regeneration are increasingly viable strategies for treating heart failure.

Abstract: Generating abundant and functional autologous cells for transplantation in such a setting, however, remains a significant challenge.

Abstract: Here, we isolated a cell population with extensive proliferation capacity and restricted cardiovascular differentiation potentials during cardiac transdifferentiation of mouse fibroblasts.

Abstract: These induced expandable cardiovascular progenitor cells (ieCPCs) proliferated extensively for more than 18 passages in chemically defined conditions, with 10(5) starting fibroblasts robustly producing 10(16) ieCPCs.

Abstract: ieCPCs expressed cardiac signature genes and readily differentiated into functional cardiomyocytes (CMs), endothelial cells (ECs), and smooth muscle cells (SMCs) in vitro, even after long-term expansion.

Abstract: When transplanted into mouse hearts following myocardial infarction, ieCPCs spontaneously differentiated into CMs, ECs, and SMCs and improved cardiac function for up to 12 weeks after transplantation.

Abstract: Thus, ieCPCs are a powerful system to study cardiovascular specification and provide strategies for regenerative medicine in the heart.


Thyroid hormone inhibits lung fibrosis in mice by improving epithelial mitochondrial function

Abstract: Thyroid hormone (TH) is critical for the maintenance of cellular homeostasis during stress responses, but its role in lung fibrosis is unknown.

Abstract: Here we found that the activity and expression of iodothyronine deiodinase 2 (DIO2), an enzyme that activates TH, were higher in lungs from patients with idiopathic pulmonary fibrosis than in control individuals and were correlated with disease severity.

Abstract: We also found that Dio2-knockout mice exhibited enhanced bleomycin-induced lung fibrosis.

Abstract: Aerosolized TH delivery increased survival and resolved fibrosis in two models of pulmonary fibrosis in mice (intratracheal bleomycin and inducible TGF-β1).

Abstract: Sobetirome, a TH mimetic, also blunted bleomycin-induced lung fibrosis.

Abstract: After bleomycin-induced injury, TH promoted mitochondrial biogenesis, improved mitochondrial bioenergetics and attenuated mitochondria-regulated apoptosis in alveolar epithelial cells both in vivo and in vitro.

Abstract: TH did not blunt fibrosis in Ppargc1a- or Pink1-knockout mice, suggesting dependence on these pathways.

Abstract: We conclude that the antifibrotic properties of TH are associated with protection of alveolar epithelial cells and restoration of mitochondrial function and that TH may thus represent a potential therapy for pulmonary fibrosis.


Tenomodulin promotes human adipocyte differentiation and beneficial visceral adipose tissue expansion.

Abstract: Proper regulation of energy storage in adipose tissue is crucial for maintaining insulin sensitivity and molecules contributing to this process have not been fully revealed.

Abstract: Here we show that type II transmembrane protein tenomodulin (TNMD) is upregulated in adipose tissue of insulin-resistant versus insulin-sensitive individuals, who were matched for body mass index (BMI).

Abstract: TNMD expression increases in human preadipocytes during differentiation, whereas silencing TNMD blocks adipogenesis.

Abstract: Upon high-fat diet feeding, transgenic mice overexpressing Tnmd develop increased epididymal white adipose tissue (eWAT) mass, and preadipocytes derived from Tnmd transgenic mice display greater proliferation, consistent with elevated adipogenesis.

Abstract: In Tnmd transgenic mice, lipogenic genes are upregulated in eWAT, as is Ucp1 in brown fat, while liver triglyceride accumulation is attenuated.

Abstract: Despite expanded eWAT, transgenic animals display improved systemic insulin sensitivity, decreased collagen deposition and inflammation in eWAT, and increased insulin stimulation of Akt phosphorylation.

Abstract: Our data suggest that TNMD acts as a protective factor in visceral adipose tissue to alleviate insulin resistance in obesity.


Evidence for a limit to human lifespan

Abstract: Driven by technological progress, human life expectancy has increased greatly since the nineteenth century.

Abstract: Demographic evidence has revealed an ongoing reduction in old-age mortality and a rise of the maximum age at death, which may gradually extend human longevity.

Abstract: Together with observations that lifespan in various animal species is flexible and can be increased by genetic or pharmaceutical intervention, these results have led to suggestions that longevity may not be subject to strict, species-specific genetic constraints.

Abstract: Here, by analysing global demographic data, we show that improvements in survival with age tend to decline after age 100, and that the age at death of the world’s oldest person has not increased since the 1990s.

Abstract: Our results strongly suggest that the maximum lifespan of humans is fixed and subject to natural constraints.


Effect of Oral Semaglutide Compared With Placebo and Subcutaneous Semaglutide on Glycemic Control in Patients With Type 2 Diabetes: A Randomized Clinical Trial

Abstract: Importance Glucagon-like peptide-1 (GLP-1) receptor agonists are effective therapies for the treatment of type 2 diabetes and are all currently available as an injection.

Abstract: Objectives To compare the effects of oral semaglutide with placebo (primary) and open-label subcutaneous semaglutide (secondary) on glycemic control in patients with type 2 diabetes.

Abstract: Design, Setting, and Patients Phase 2, randomized, parallel-group, dosage-finding, 26-week trial with 5-week follow-up at 100 sites (hospital clinics, general practices, and clinical research centers) in 14 countries conducted between December 2013 and December 2014.

Abstract: Of 1106 participants assessed, 632 with type 2 diabetes and insufficient glycemic control using diet and exercise alone or a stable dose of metformin were randomized.

Abstract: Randomization was stratified by metformin use.

Abstract: Interventions Once-daily oral semaglutide of 2.5 mg (n = 70), 5 mg (n = 70), 10 mg (n = 70), 20 mg (n = 70), 40-mg 4-week dose escalation (standard escalation; n = 71), 40-mg 8-week dose escalation (slow escalation; n = 70), 40-mg 2-week dose escalation (fast escalation, n = 70), oral placebo (n = 71; double-blind) or once-weekly subcutaneous semaglutide of 1.0 mg (n = 70) for 26 weeks.

Abstract: Main Outcomes and Measures The primary end point was change in hemoglobing A1c (HbA1c) from baseline to week 26.

Abstract: Secondary end points included change from baseline in body weight and adverse events.

Abstract: Results Baseline characteristics were comparable across treatment groups.

Abstract: Of the 632 randomized patients (mean age, 57.1 years [SD, 10.6]; men, 395 (62.7%); diabetes duration, 6.3 years [SD, 5.2]; body weight, 92.3 kg [SD, 16.8]; BMI, 31.7 [SD, 4.3]), 583 (92%) completed the trial.

Abstract: Mean change in HbA1c level from baseline to week 26 decreased with oral semaglutide (dosage-dependent range, −0.7% to −1.9%) and subcutaneous semaglutide (−1.9%) and placebo (−0.3%); oral semaglutide reductions were significant vs placebo (dosage-dependent estimated treatment difference [ETD] range for oral semaglutide vs placebo, –0.4% to –1.6%; P = .01 for 2.5 mg, <.001 for all other dosages).

Abstract: Reductions in body weight were greater with oral semaglutide (dosage-dependent range, −2.1 kg to −6.9 kg) and subcutaneous semaglutide (−6.4 kg) vs placebo (−1.2 kg), and significant for oral semaglutide dosages of 10 mg or more vs placebo (dosage-dependent ETD range, –0.9 to –5.7 kg; P < .001).

Abstract: Adverse events were reported by 63% to 86% (371 of 490 patients) in the oral semaglutide groups, 81% (56 of 69 patients) in the subcutaneous semaglutide group, and 68% (48 of 71 patients) in the placebo group; mild to moderate gastrointestinal events were most common.

Abstract: Conclusions and Relevance Among patients with type 2 diabetes, oral semaglutide resulted in better glycemic control than placebo over 26 weeks.

Abstract: These findings support phase 3 studies to assess longer-term and clinical outcomes, as well as safety.

Abstract: Trial Registration clinicaltrials.gov Identifier: NCT01923181


Fast and accurate short read alignment with Burrows–Wheeler transform

Abstract: MOTIVATION The enormous amount of short reads generated by the new DNA sequencing technologies call for the development of fast and accurate read alignment programs.

Abstract: A first generation of hash table-based methods has been developed, including MAQ, which is accurate, feature rich and fast enough to align short reads from a single individual.

Abstract: However, MAQ does not support gapped alignment for single-end reads, which makes it unsuitable for alignment of longer reads where indels may occur frequently.

Abstract: The speed of MAQ is also a concern when the alignment is scaled up to the resequencing of hundreds of individuals.

Abstract: RESULTS We implemented Burrows-Wheeler Alignment tool (BWA), a new read alignment package that is based on backward search with Burrows-Wheeler Transform (BWT), to efficiently align short sequencing reads against a large reference sequence such as the human genome, allowing mismatches and gaps.

Abstract: BWA supports both base space reads, e.g. from Illumina sequencing machines, and color space reads from AB SOLiD machines.

Abstract: Evaluations on both simulated and real data suggest that BWA is approximately 10-20x faster than MAQ, while achieving similar accuracy.

Abstract: In addition, BWA outputs alignment in the new standard SAM (Sequence Alignment/Map) format.

Abstract: Variant calling and other downstream analyses after the alignment can be achieved with the open source SAMtools software package.

Abstract: AVAILABILITY http://maq.sourceforge.net.


The financial cost of doctors emigrating from sub-Saharan Africa: human capital analysis

Abstract: OBJECTIVE To estimate the lost investment of domestically educated doctors migrating from sub-Saharan African countries to Australia, Canada, the United Kingdom, and the United States.

Abstract: DESIGN Human capital cost analysis using publicly accessible data.

Abstract: SETTINGS Sub-Saharan African countries.

Abstract: PARTICIPANTS Nine sub-Saharan African countries with an HIV prevalence of 5% or greater or with more than one million people with HIV/AIDS and with at least one medical school (Ethiopia, Kenya, Malawi, Nigeria, South Africa, Tanzania, Uganda, Zambia, and Zimbabwe), and data available on the number of doctors practising in destination countries.

Abstract: MAIN OUTCOME MEASURES The financial cost of educating a doctor (through primary, secondary, and medical school), assuming that migration occurred after graduation, using current country specific interest rates for savings converted to US dollars; cost according to the number of source country doctors currently working in the destination countries; and savings to destination countries of receiving trained doctors.

Abstract: RESULTS In the nine source countries the estimated government subsidised cost of a doctor's education ranged from $21,000 (£13,000; €15,000) in Uganda to $58,700 in South Africa.

Abstract: The overall estimated loss of returns from investment for all doctors currently working in the destination countries was $2.17bn (95% confidence interval 2.13bn to 2.21bn), with costs for each country ranging from $2.16m (1.55m to 2.78m) for Malawi to $1.41bn (1.38bn to 1.44bn) for South Africa.

Abstract: The ratio of the estimated compounded lost investment over gross domestic product showed that Zimbabwe and South Africa had the largest losses.

Abstract: The benefit to destination countries of recruiting trained doctors was largest for the United Kingdom ($2.7bn) and United States ($846m).

Abstract: CONCLUSIONS Among sub-Saharan African countries most affected by HIV/AIDS, lost investment from the emigration of doctors is considerable.

Abstract: Destination countries should consider investing in measurable training for source countries and strengthening of their health systems.


Chromatin modifying enzymes as modulators of reprogramming

Abstract: Generation of induced pluripotent stem cells (iPSCs) by somatic cell reprogramming involves global epigenetic remodelling.

Abstract: Whereas several proteins are known to regulate chromatin marks associated with the distinct epigenetic states of cells before and after reprogramming, the role of specific chromatin-modifying enzymes in reprogramming remains to be determined.

Abstract: To address how chromatin-modifying proteins influence reprogramming, we used short hairpin RNAs (shRNAs) to target genes in DNA and histone methylation pathways, and identified positive and negative modulators of iPSC generation.

Abstract: Whereas inhibition of the core components of the polycomb repressive complex 1 and 2, including the histone 3 lysine 27 methyltransferase EZH2, reduced reprogramming efficiency, suppression of SUV39H1, YY1 and DOT1L enhanced reprogramming.

Abstract: Specifically, inhibition of the H3K79 histone methyltransferase DOT1L by shRNA or a small molecule accelerated reprogramming, significantly increased the yield of iPSC colonies, and substituted for KLF4 and c-Myc (also known as MYC).

Abstract: Inhibition of DOT1L early in the reprogramming process is associated with a marked increase in two alternative factors, NANOG and LIN28, which play essential functional roles in the enhancement of reprogramming.

Abstract: Genome-wide analysis of H3K79me2 distribution revealed that fibroblast-specific genes associated with the epithelial to mesenchymal transition lose H3K79me2 in the initial phases of reprogramming.

Abstract: DOT1L inhibition facilitates the loss of this mark from genes that are fated to be repressed in the pluripotent state.

Abstract: These findings implicate specific chromatin-modifying enzymes as barriers to or facilitators of reprogramming, and demonstrate how modulation of chromatin-modifying enzymes can be exploited to more efficiently generate iPSCs with fewer exogenous transcription factors.


Natural variation in the parameters of innate immune cells is preferentially driven by genetic factors

Abstract: The quantification and characterization of circulating immune cells provide key indicators of human health and disease.

Abstract: To identify the relative effects of environmental and genetic factors on variation in the parameters of innate and adaptive immune cells in homeostatic conditions, we combined standardized flow cytometry of blood leukocytes and genome-wide DNA genotyping of 1,000 healthy, unrelated people of Western European ancestry.

Abstract: We found that smoking, together with age, sex and latent infection with cytomegalovirus, were the main non-genetic factors that affected variation in parameters of human immune cells.

Abstract: Genome-wide association studies of 166 immunophenotypes identified 15 loci that showed enrichment for disease-associated variants.

Abstract: Finally, we demonstrated that the parameters of innate cells were more strongly controlled by genetic variation than were those of adaptive cells, which were driven by mainly environmental exposure.

Abstract: Our data establish a resource that will generate new hypotheses in immunology and highlight the role of innate immunity in susceptibility to common autoimmune diseases.

Abstract: Both environmental factors and genetic factors influence human immunity.

Abstract: Albert and colleagues leverage data from the Milieu Intérieur Consortium to comprehensively describe the effects of lifestyle, environment and genetics on human innate and adaptive immunity.


The transcriptional program, functional heterogeneity, and clinical targeting of mast cells

Abstract: Mast cells are unique tissue-resident immune cells that express an array of receptors that can be activated by several extracellular cues, including antigen-immunoglobulin E (IgE) complexes, bacteria, viruses, cytokines, hormones, peptides, and drugs.

Abstract: Mast cells constitute a small population in tissues, but their extraordinary ability to respond rapidly by releasing granule-stored and newly made mediators underpins their importance in health and disease.

Abstract: In this review, we document the biology of mast cells and introduce new concepts and opinions regarding their role in human diseases beyond IgE-mediated allergic responses and antiparasitic functions.

Abstract: We bring to light recent discoveries and developments in mast cell research, including regulation of mast cell functions, differentiation, survival, and novel mouse models.

Abstract: Finally, we highlight the current and future opportunities for therapeutic intervention of mast cell functions in inflammatory diseases.


LIN28B enhanced tumorigenesis in an autochthonous KRASG12V-driven lung carcinoma mouse model

Abstract: LIN28B is a RNA-binding protein regulating predominantly let-7 microRNAs with essential functions in inflammation, wound healing, embryonic stem cells, and cancer.

Abstract: LIN28B expression is associated with tumor initiation, progression, resistance, and poor outcome in several solid cancers, including lung cancer.

Abstract: However, the functional role of LIN28B, especially in non-small cell lung adenocarcinomas, remains elusive.

Abstract: Here, we investigated the effects of LIN28B expression on lung tumorigenesis using LIN28B transgenic overexpression in an autochthonous KRASG12V-driven mouse model.

Abstract: We found that LIN28B overexpression significantly increased the number of CD44+/CD326+ tumor cells, upregulated VEGF-A and miR-21 and promoted tumor angiogenesis and epithelial-to-mesenchymal transition (EMT) accompanied by enhanced AKT phosphorylation and nuclear translocation of c-MYC.

Abstract: Moreover, LIN28B accelerated tumor initiation and enhanced proliferation which led to a shortened overall survival.

Abstract: In addition, we analyzed lung adenocarcinomas of the Cancer Genome Atlas (TCGA) and found LIN28B expression in 24% of KRAS-mutated cases, which underscore the relevance of our model.


Cost-effectiveness comparison between topical silver sulfadiazine and enclosed silver dressing for partial-thickness burn treatment.

Abstract: The standard treatment of partial-thickness burns includes topical silver products such as silver sulfadiazine (SSD) cream and enclosed dressings including silver-impregnated foam (Mepilex Ag; Molnlycke Health Care, Gothenburg, Sweden) and silver-laden sheets (Aquacel Ag; ConvaTec, Skillman, NJ).

Abstract: The current state of health care is limited by resources, with an emphasis on evidence-based outcomes and cost-effective treatments.

Abstract: This study includes a decision analysis with an incremental cost-utility ratio comparing enclosed silver dressings with SSD in partial-thickness burn patients with TBSA less than 20%.

Abstract: A comprehensive literature review was conducted to identify clinically relevant health states in partial-thickness burn patients.

Abstract: These health states include successful healing, infection, and noninfected delayed healing requiring either surgery or conservative management.

Abstract: The probabilities of these health states were combined with Medicare CPT reimbursement codes (cost) and patient-derived utilities to fit into the decision model.

Abstract: Utilities were obtained using a visual analog scale during patient interviews.

Abstract: Expected cost and quality-adjusted life years (QALYs) were calculated using the roll-back method.

Abstract: The incremental cost-utility ratio for enclosed silver dressing relative to SSD was $40,167.99/QALY.

Abstract: One-way sensitivity analysis of complication rates confirmed robustness of the model.

Abstract: Assuming a maximum willingness to pay $50,000/QALY, the complication rate for SSD must be 22% or higher for enclosed silver dressing to be cost effective.

Abstract: By varying complication rates for SSD and enclosed silver dressings, the two-way sensitivity analysis demonstrated the cost effectiveness of using enclosed silver dressing at the majority of complication rates for both treatment modalities.

Abstract: Enclosed silver dressings are a cost-effective means of treating partial thickness burns.


Randomized clinical study of Hydrofiber dressing with silver or silver sulfadiazine in the management of partial-thickness burns.

Abstract: This prospective, randomized study compared protocols of care using either AQUACEL Ag Hydrofiber (ConvaTec, a Bristol-Myers Squibb company, Skillman, NJ) dressing with silver (n = 42) or silver sulfadiazine (n = 42) for up to 21 days in the management of partial-thickness burns covering 5% to 40% body surface area (BSA).

Abstract: AQUACEL Ag dressing was associated with less pain and anxiety during dressing changes, less burning and stinging during wear, fewer dressing changes, less nursing time, and fewer procedural medications.

Abstract: Silver sulfadiazine was associated with greater flexibility and ease of movement.

Abstract: Adverse events, including infection, were comparable between treatment groups.

Abstract: The AQUACEL Ag dressing protocol tended to have lower total treatment costs (Dollars 1040 vs. Dollars 1180) and a greater rate of re-epithelialization (73.8% vs 60.0%), resulting in cost-effectiveness per burn healed of Dollars 1,409.06 for AQUACEL Ag dressing and Dollars 1,967.95 for silver sulfadiazine.

Abstract: A protocol of care with AQUACEL(R) Ag provided clinical and economic benefits compared with silver sulfadiazine in patients with partial-thickness burns.


Hepatitis C virus and blood transfusion: past and present risks.

Abstract: The risk of HCV transmission by blood and blood products has been greatly reduced since the early 1980's.

Abstract: Selection of non-remunerated donors, donor selection to prevent HIV transmission, initial surrogate testing in some regions, and introduction of anti-HCV testing have all contributed to this.

Abstract: ALT surrogate testing has become obsolete since the introduction of anti-HCV testing.

Abstract: The residual risk of HCV transmission due to donations in the anti-HCV window period at present is about 1 in 100 000 transfusions of cellular products, and transmission of HCV by plasma products treated with modern inactivation methods such as solvent-detergent treatment, has not been reported.

Abstract: Hemovigilance programmes, which are presently being installed, will provide more data on the safety of blood transfusion.

Abstract: Introduction of HCV nucleic amplification technology (NAT) as a quality control of manufacturing pools for plasma products or as a form of blood donor screening by minipools is anticipated in many European countries for the coming year.

Abstract: Given industrial developments, NAT testing of individual blood donations may become available within the next 2 years.

Abstract: HCV NAT testing will further annihilate the residual risk, and the cost-effectiveness will become relatively low in comparison with other public health measures.


p62/SQSTM1 by Binding to Vitamin D Receptor Inhibits Hepatic Stellate Cell Activity, Fibrosis, and Liver Cancer.

Abstract: Hepatic stellate cells (HSCs) play critical roles in liver fibrosis and hepatocellular carcinoma (HCC).

Abstract: Vitamin D receptor (VDR) activation in HSCs inhibits liver inflammation and fibrosis.

Abstract: We found that p62/SQSTM1, a protein upregulated in liver parenchymal cells but downregulated in HCC-associated HSCs, negatively controls HSC activation.

Abstract: Total body or HSC-specific p62 ablation potentiates HSCs and enhances inflammation, fibrosis, and HCC progression.

Abstract: p62 directly interacts with VDR and RXR promoting their heterodimerization, which is critical for VDR:RXR target gene recruitment.

Abstract: Loss of p62 in HSCs impairs the repression of fibrosis and inflammation by VDR agonists.

Abstract: This demonstrates that p62 is a negative regulator of liver inflammation and fibrosis through its ability to promote VDR signaling in HSCs, whose activation supports HCC.


Developing criteria for evaluation of geroprotectors as a key stage toward translation to the clinic

Abstract: In the coming decades, a massive shift in the aging segment of the population will have major social and economic consequences around the world.

Abstract: One way to offset this increase is to expedite the development of geroprotectors, substances that slow aging, repair age-associated damage and extend healthy lifespan, or healthspan.

Abstract: While over 200 geroprotectors are now reported in model organisms and some are in human use for specific disease indications, the path toward determining whether they affect aging in humans remains obscure.

Abstract: Translation to the clinic is hampered by multiple issues including absence of a common set of criteria to define, select, and classify these substances, given the complexity of the aging process and their enormous diversity in mechanism of action.

Abstract: Translational research efforts would benefit from the formation of a scientific consensus on the following: the definition of 'geroprotector', the selection criteria for geroprotectors, a comprehensive classification system, and an analytical model.

Abstract: Here, we review current approaches to selection and put forth our own suggested selection criteria.

Abstract: Standardizing selection of geroprotectors will streamline discovery and analysis of new candidates, saving time and cost involved in translation to clinic.


β-Catenin Signaling and Roles in Liver Homeostasis, Injury, and Tumorigenesis.

Abstract: β-catenin (encoded by CTNNB1) is a subunit of the cell surface cadherin protein complex that acts as an intracellular signal transducer in the WNT signaling pathway; alterations in its activity have been associated with the development of hepatocellular carcinoma and other liver diseases.

Abstract: Other than WNT, additional signaling pathways also can converge at β-catenin.

Abstract: β-catenin also interacts with transcription factors such as T-cell factor, forkhead box protein O, and hypoxia inducible factor 1α to regulate the expression of target genes.

Abstract: We discuss the role of β-catenin in metabolic zonation of the adult liver.

Abstract: β-catenin also regulates the expression of genes that control metabolism of glucose, nutrients, and xenobiotics; alterations in its activity may contribute to the pathogenesis of nonalcoholic steatohepatitis.

Abstract: Alterations in β-catenin signaling may lead to activation of hepatic stellate cells, which is required for fibrosis.

Abstract: Many hepatic tumors such as hepatocellular adenomas, hepatocellular cancers, and hepatoblastomas have mutations in CTNNB1 that result in constitutive activation of β-catenin, so this molecule could be a therapeutic target.

Abstract: We discuss how alterations in β-catenin activity contribute to liver disease and how these might be used in diagnosis and prognosis, as well as in the development of therapeutics.


The global burden of dengue: an analysis from the Global Burden of Disease Study 2013.

Abstract: BACKGROUND Dengue is the most common arbovirus infection globally, but its burden is poorly quantified.

Abstract: We estimated dengue mortality, incidence, and burden for the Global Burden of Disease Study 2013.

Abstract: METHODS We modelled mortality from vital registration, verbal autopsy, and surveillance data using the Cause of Death Ensemble Modelling tool.

Abstract: We modelled incidence from officially reported cases, and adjusted our raw estimates for under-reporting based on published estimates of expansion factors.

Abstract: In total, we had 1780 country-years of mortality data from 130 countries, 1636 country-years of dengue case reports from 76 countries, and expansion factor estimates for 14 countries.

Abstract: FINDINGS We estimated an average of 9221 dengue deaths per year between 1990 and 2013, increasing from a low of 8277 (95% uncertainty estimate 5353-10 649) in 1992, to a peak of 11 302 (6790-13 722) in 2010.

Abstract: This yielded a total of 576 900 (330 000-701 200) years of life lost to premature mortality attributable to dengue in 2013.

Abstract: The incidence of dengue increased greatly between 1990 and 2013, with the number of cases more than doubling every decade, from 8·3 million (3·3 million-17·2 million) apparent cases in 1990, to 58·4 million (23·6 million-121·9 million) apparent cases in 2013.

Abstract: When accounting for disability from moderate and severe acute dengue, and post-dengue chronic fatigue, 566 000 (186 000-1 415 000) years lived with disability were attributable to dengue in 2013.

Abstract: Considering fatal and non-fatal outcomes together, dengue was responsible for 1·14 million (0·73 million-1·98 million) disability-adjusted life-years in 2013.

Abstract: INTERPRETATION Although lower than other estimates, our results offer more evidence that the true symptomatic incidence of dengue probably falls within the commonly cited range of 50 million to 100 million cases per year.

Abstract: Our mortality estimates are lower than those presented elsewhere and should be considered in light of the totality of evidence suggesting that dengue mortality might, in fact, be substantially higher.

Abstract: FUNDING Bill & Melinda Gates Foundation.


Formin-mediated actin polymerization at cell–cell junctions stabilizes E-cadherin and maintains monolayer integrity during wound repair

Abstract: Cadherin-mediated cell-cell adhesion is required for epithelial tissue integrity in homeostasis, during development, and in tissue repair.

Abstract: E-cadherin stability depends on F-actin, but the mechanisms regulating actin polymerization at cell-cell junctions remain poorly understood.

Abstract: Here we investigated a role for formin-mediated actin polymerization at cell-cell junctions.

Abstract: We identify mDia1 and Fmnl3 as major factors enhancing actin polymerization and stabilizing E-cadherin at epithelial junctions.

Abstract: Fmnl3 localizes to adherens junctions downstream of Src and Cdc42 and its depletion leads to a reduction in F-actin and E-cadherin at junctions and a weakening of cell-cell adhesion.

Abstract: Of importance, Fmnl3 expression is up-regulated and junctional localization increases during collective cell migration.

Abstract: Depletion of Fmnl3 or mDia1 in migrating monolayers results in dissociation of leader cells and impaired wound repair.

Abstract: In summary, our results show that formin activity at epithelial cell-cell junctions is important for adhesion and the maintenance of epithelial cohesion during dynamic processes, such as wound repair.


A mechanically active heterotypic E-cadherin/N-cadherin adhesion enables fibroblasts to drive cancer cell invasion

Abstract: Cancer-associated fibroblasts (CAFs) promote tumour invasion and metastasis.

Abstract: We show that CAFs exert a physical force on cancer cells that enables their collective invasion.

Abstract: Force transmission is mediated by a heterophilic adhesion involving N-cadherin at the CAF membrane and E-cadherin at the cancer cell membrane.

Abstract: This adhesion is mechanically active; when subjected to force it triggers β-catenin recruitment and adhesion reinforcement dependent on α-catenin/vinculin interaction.

Abstract: Impairment of E-cadherin/N-cadherin adhesion abrogates the ability of CAFs to guide collective cell migration and blocks cancer cell invasion.

Abstract: N-cadherin also mediates repolarization of the CAFs away from the cancer cells.

Abstract: In parallel, nectins and afadin are recruited to the cancer cell/CAF interface and CAF repolarization is afadin dependent.

Abstract: Heterotypic junctions between CAFs and cancer cells are observed in patient-derived material.

Abstract: Together, our findings show that a mechanically active heterophilic adhesion between CAFs and cancer cells enables cooperative tumour invasion.


Long non-coding RNA HOTTIP promotes BCL-2 expression and induces chemoresistance in small cell lung cancer by sponging miR-216a

Abstract: Despite progress in treatment of small cell lung cancer (SCLC), its multidrug chemoresistance and poor prognosis still remain.

Abstract: Recently, we globally assessed long non-coding RNAs (lncRNAs) for contributions to SCLC chemoresistance using microarray data, in vitro and in vivo assays.

Abstract: Here we reported that HOTTIP, encoding a lncRNA that is frequently amplified in SCLC, was associated with SCLC cell chemosensitivity, proliferation, and poor prognosis of SCLC patients.

Abstract: Moreover, mechanistic investigations showed that HOTTIP functioned as an oncogene in SCLC progression by binding miR-216a and abrogating its tumor-suppressive function in this setting.

Abstract: On the other hand, HOTTIP increased the expression of anti-apoptotic factor BCL-2, another important target gene of miR-216a, and jointly enhanced chemoresistance of SCLC by regulating BCL-2.

Abstract: Taken together, our study established a role for HOTTIP in SCLC progression and chemoresistance suggest its candidacy as a new diagnostic and prognostic biomarker for clinical management of SCLC.


GSDS 2.0: an upgraded gene feature visualization server

Abstract: UNLABELLED : Visualizing genes' structure and annotated features helps biologists to investigate their function and evolution intuitively.

Abstract: The Gene Structure Display Server (GSDS) has been widely used by more than 60 000 users since its first publication in 2007.

Abstract: Here, we reported the upgraded GSDS 2.0 with a newly designed interface, supports for more types of annotation features and formats, as well as an integrated visual editor for editing the generated figure.

Abstract: Moreover, a user-specified phylogenetic tree can be added to facilitate further evolutionary analysis.

Abstract: The full source code is also available for downloading.

Abstract: AVAILABILITY AND IMPLEMENTATION Web server and source code are freely available at http://gsds.cbi.pku.edu.cn.

Abstract: CONTACT gaog@mail.cbi.pku.edu.cn or gsds@mail.cbi.pku.edu.cn   SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online.


SLP-65 regulates immunoglobulin light chain gene recombination through the PI(3)K-PKB-Foxo pathway

Abstract: Although the essential role of the adaptor protein SLP-65 in pre-B cell differentiation is established, the molecular mechanism underlying its function is poorly understood.

Abstract: In this study, we uncover a link between SLP-65–dependent signaling and the phosphoinositide-3-OH kinase (PI(3)K)–protein kinase B (PKB)–Foxo pathway.

Abstract: We show that the forkhead box transcription factor Foxo3a promotes light chain rearrangement in pre-B cells.

Abstract: Our data suggest that PKB suppresses light chain recombination by phosphorylating Foxo proteins, whereas reconstitution of SLP-65 function counteracts PKB activation and promotes Foxo3a and Foxo1 activity in pre-B cells.

Abstract: Together, these data illuminate a molecular function of SLP-65 and identify a key role for Foxo proteins in the regulation of light chain recombination, receptor editing and B cell selection.


A case-mix classification system for explaining healthcare costs using administrative data in Italy.

Abstract: BACKGROUND The Italian National Health Service (NHS) provides universal coverage to all citizens, granting primary and hospital care with a copayment system for outpatient and drug services.

Abstract: Financing of Local Health Trusts (LHTs) is based on a capitation system adjusted only for age, gender and area of residence.

Abstract: We applied a risk-adjustment system (Johns Hopkins Adjusted Clinical Groups System, ACG® System) in order to explain health care costs using routinely collected administrative data in the Veneto Region (North-eastern Italy).

Abstract: METHODS All residents in the Veneto Region were included in the study.

Abstract: The ACG system was applied to classify the regional population based on the following information sources for the year 2015: Hospital Discharges, Emergency Room visits, Chronic disease registry for copayment exemptions, ambulatory visits, medications, the Home care database, and drug prescriptions.

Abstract: Simple linear regressions were used to contrast an age-gender model to models incorporating more comprehensive risk measures aimed at predicting health care costs.

Abstract: RESULTS A simple age-gender model explained only 8% of the variance of 2015 total costs.

Abstract: Adding diagnoses-related variables provided a 23% increase, while pharmacy based variables provided an additional 17% increase in explained variance.

Abstract: The adjusted R-squared of the comprehensive model was 6 times that of the simple age-gender model.

Abstract: CONCLUSIONS ACG System provides substantial improvement in predicting health care costs when compared to simple age-gender adjustments.

Abstract: Aging itself is not the main determinant of the increase of health care costs, which is better explained by the accumulation of chronic conditions and the resulting multimorbidity.


EpCAM-regulated intramembrane proteolysis induces a cancer stem cell-like gene signature in hepatitis B virus-infected hepatocytes.

Abstract: BACKGROUND & AIMS Hepatocytes in which the hepatitis B virus (HBV) is replicating exhibit loss of the chromatin modifying polycomb repressive complex 2 (PRC2), resulting in re-expression of specific, cellular PRC2-repressed genes.

Abstract: Epithelial cell adhesion molecule (EpCAM) is a PRC2-repressed gene, normally expressed in hepatic progenitors, but re-expressed in hepatic cancer stem cells (hCSCs).

Abstract: Herein, we investigated the functional significance of EpCAM re-expression in HBV-mediated hepatocarcinogenesis.

Abstract: METHODS Employing molecular approaches (transfections, fluorescence-activated cell sorting, immunoblotting, qRT-PCR), we investigated the role of EpCAM-regulated intramembrane proteolysis (RIP) in HBV replicating cells in vitro, and in liver tumors from HBV X/c-myc mice and chronically HBV infected patients.

Abstract: RESULTS EpCAM undergoes RIP in HBV replicating cells, activating canonical Wnt signaling.

Abstract: Transfection of Wnt-responsive plasmid expressing green fluorescent protein (GFP) identified a GFP + population of HBV replicating cells.

Abstract: These GFP+/Wnt+ cells exhibited cisplatin- and sorafenib-resistant growth resembling hCSCs, and increased expression of pluripotency genes NANOG, OCT4, SOX2, and hCSC markers BAMBI, CD44 and CD133.

Abstract: These genes are referred as EpCAM RIP and Wnt-induced hCSC-like gene signature.

Abstract: Interestingly, this gene signature is also overexpressed in liver tumors of X/c-myc bitransgenic mice.

Abstract: Clinically, a group of HBV-associated hepatocellular carcinomas was identified, exhibiting elevated expression of the hCSC-like gene signature and associated with reduced overall survival post-surgical resection.

Abstract: CONCLUSIONS The hCSC-like gene signature offers promise as prognostic tool for classifying subtypes of HBV-induced HCCs.

Abstract: Since EpCAM RIP and Wnt signaling drive expression of this hCSC-like signature, inhibition of these pathways can be explored as therapeutic strategy for this subtype of HBV-associated HCCs.

Abstract: LAY SUMMARY In this study, we provide evidence for a molecular mechanism by which chronic infection by the hepatitis B virus results in the development of poor prognosis liver cancer.

Abstract: Based on this mechanism our results suggest possible therapeutic interventions.


Parathyroid Hormone Directs Bone Marrow Mesenchymal Cell Fate.

Abstract: Intermittent PTH administration builds bone mass and prevents fractures, but its mechanism of action is unclear.

Abstract: We genetically deleted the PTH/PTHrP receptor (PTH1R) in mesenchymal stem cells using Prx1Cre and found low bone formation, increased bone resorption, and high bone marrow adipose tissue (BMAT).

Abstract: Bone marrow adipocytes traced to Prx1 and expressed classic adipogenic markers and high receptor activator of nuclear factor kappa B ligand (Rankl) expression.

Abstract: RANKL levels were also elevated in bone marrow supernatant and serum, but undetectable in other adipose depots.

Abstract: By cell sorting, Pref1+RANKL+ marrow progenitors were twice as great in mutant versus control marrow.

Abstract: Intermittent PTH administration to control mice reduced BMAT significantly.

Abstract: A similar finding was noted in male osteoporotic patients.

Abstract: Thus, marrow adipocytes exhibit osteogenic and adipogenic characteristics, are uniquely responsive to PTH, and secrete RANKL.

Abstract: These studies reveal an important mechanism for PTH's therapeutic action through its ability to direct mesenchymal cell fate.


Metformin attenuates blood-brain barrier disruption in mice following middle cerebral artery occlusion

Abstract: BACKGROUND Metformin, a widely used hypoglycemic drug, reduces stroke incidence and alleviates chronic inflammation in clinical trials.

Abstract: However, the effect of metformin in ischemic stroke is unclear.

Abstract: Here, we investigated the effect of metformin on ischemic stroke in mice and further explored the possible underlying mechanisms.

Abstract: METHODS Ninety-eight adult male CD-1 mice underwent 90-minute transient middle cerebral artery occlusion (tMCAO).

Abstract: Metformin (200 mg/kg) was administrated for up to 14 days.

Abstract: Neurobehavioral outcomes, brain infarct volume, inflammatory factors, blood-brain barrier (BBB) permeability and AMPK signaling pathways were evaluated following tMCAO.

Abstract: Oxygen glucose deprivation was performed on bEND.3 cells to explore the mechanisms of metformin in inhibiting inflammatory signaling pathways.

Abstract: RESULTS Infarct volume was reduced in metformin-treated mice compared to the control group following tMCAO (P < 0.05).

Abstract: Neurobehavioral outcomes were greatly improved in metformin-treated mice (P < 0.05).

Abstract: MPO+ cells, Gr1+ cells, MPO activity and BBB permeability were decreased after metformin administration (P < 0.05).

Abstract: In addition, metformin activated AMPK phosphorylation, inhibited NF-κB activation, down-regulated cytokine (IL-1β, IL-6, TNF-α) and ICAM-1 expression following tMCAO (P < 0.05).

Abstract: Furthermore, metformin activated AMPK signaling pathway and alleviated oxygen-glucose deprivation-induced ICAM-1 expression in bEND.3 cells (P < 0.05).

Abstract: Compound C, a selective AMPK inhibitor, eliminated this promotional effect.

Abstract: CONCLUSIONS Metformin down-regulated ICAM-1 in an AMPK-dependent manner, which could effectively prevent ischemia-induced brain injury by alleviating neutrophil infiltration, suggesting that metformin is a promising therapeutic agent in stroke therapy.


Epstein-Barr Virus nuclear antigen 1 (EBNA1) confers resistance to apoptosis in EBV-positive B-lymphoma cells through up-regulation of survivin.

Abstract: Resistance to apoptosis is an important component of the overall mechanism which drives the tumorigenic process.

Abstract: EBV is a ubiquitous human gamma-herpesvirus which preferentially establishes latent infection in viral infected B-lymphocytes.

Abstract: EBNA1 is typically expressed in most forms of EBV-positive malignancies and is important for replication of the latent episome in concert with replication of the host cells.

Abstract: Here, we investigate the effects of EBNA1 on survivin up-regulation in EBV-infected human B-lymphoma cells.

Abstract: We present evidence which demonstrates that EBNA1 forms a complex with Sp1 or Sp1-like proteins bound to their cis-element at the survivin promoter.

Abstract: This enhances the activity of the complex and up-regulates survivin.

Abstract: Knockdown of survivin and EBNA1 showed enhanced apoptosis in infected cells and thus supports a role for EBNA1 in suppressing apoptosis in EBV-infected cells.

Abstract: Here, we suggest that EBV encoded EBNA1 can contribute to the oncogenic process by up-regulating the apoptosis suppressor protein, survivin in EBV-associated B-lymphoma cells.


Microfluidic platform to evaluate migration of cells from patients with DYT1 dystonia.

Abstract: BACKGROUND Microfluidic platforms for quantitative evaluation of cell biologic processes allow low cost and time efficient research studies of biological and pathological events, such as monitoring cell migration by real-time imaging.

Abstract: In healthy and disease states, cell migration is crucial in development and wound healing, as well as to maintain the body's homeostasis.

Abstract: NEW METHOD The microfluidic chambers allow precise measurements to investigate whether fibroblasts carrying a mutation in the TOR1A gene, underlying the hereditary neurologic disease--DYT1 dystonia, have decreased migration properties when compared to control cells.

Abstract: RESULTS We observed that fibroblasts from DYT1 patients showed abnormalities in basic features of cell migration, such as reduced velocity and persistence of movement.

Abstract: COMPARISON WITH EXISTING METHOD The microfluidic method enabled us to demonstrate reduced polarization of the nucleus and abnormal orientation of nuclei and Golgi inside the moving DYT1 patient cells compared to control cells, as well as vectorial movement of single cells.

Abstract: CONCLUSION We report here different assays useful in determining various parameters of cell migration in DYT1 patient cells as a consequence of the TOR1A gene mutation, including a microfluidic platform, which provides a means to evaluate real-time vectorial movement with single cell resolution in a three-dimensional environment.


Sugar-sweetened beverages and weight gain in children and adults: a systematic review and meta-analysis.

Abstract: BACKGROUND The relation between sugar-sweetened beverages (SSBs) and body weight remains controversial.

Abstract: OBJECTIVE We conducted a systematic review and meta-analysis to summarize the evidence in children and adults.

Abstract: DESIGN We searched PubMed, EMBASE, and Cochrane databases through March 2013 for prospective cohort studies and randomized controlled trials (RCTs) that evaluated the SSB-weight relation.

Abstract: Separate meta-analyses were conducted in children and adults and for cohorts and RCTs by using random- and fixed-effects models.

Abstract: RESULTS Thirty-two original articles were included in our meta-analyses: 20 in children (15 cohort studies, n = 25,745; 5 trials, n = 2772) and 12 in adults (7 cohort studies, n = 174,252; 5 trials, n = 292).

Abstract: In cohort studies, one daily serving increment of SSBs was associated with a 0.06 (95% CI: 0.02, 0.10) and 0.05 (95% CI: 0.03, 0.07)-unit increase in BMI in children and 0.22 kg (95% CI: 0.09, 0.34 kg) and 0.12 kg (95% CI: 0.10, 0.14 kg) weight gain in adults over 1 y in random- and fixed-effects models, respectively.

Abstract: RCTs in children showed reductions in BMI gain when SSBs were reduced [random and fixed effects: -0.17 (95% CI: -0.39, 0.05) and -0.12 (95% CI: -0.22, -0.2)], whereas RCTs in adults showed increases in body weight when SSBs were added (random and fixed effects: 0.85 kg; 95% CI: 0.50, 1.20 kg).

Abstract: Sensitivity analyses of RCTs in children showed more pronounced benefits in preventing weight gain in SSB substitution trials (compared with school-based educational programs) and among overweight children (compared with normal-weight children).

Abstract: CONCLUSION Our systematic review and meta-analysis of prospective cohort studies and RCTs provides evidence that SSB consumption promotes weight gain in children and adults.


The tumorigenic potential of pluripotent stem cells: What can we do to minimize it?

Abstract: Human pluripotent stem cells (hPSCs) have the potential to fundamentally change the way that we go about treating and understanding human disease.

Abstract: Despite this extraordinary potential, these cells also have an innate capability to form tumors in immunocompromised individuals when they are introduced in their pluripotent state.

Abstract: Although current therapeutic strategies involve transplantation of only differentiated hPSC derivatives, there is still a concern that transplanted cell populations could contain a small percentage of cells that are not fully differentiated.

Abstract: In addition, these cells have been frequently reported to acquire genetic alterations that, in some cases, are associated with certain types of human cancers.

Abstract: Here, we try to separate the panic from reality and rationally evaluate the true tumorigenic potential of these cells.

Abstract: We also discuss a recent study examining the effect of culture conditions on the genetic integrity of hPSCs.

Abstract: Finally, we present a set of sensible guidelines for minimizing the tumorigenic potential of hPSC-derived cells.

Abstract: © 2016 The Authors.

Abstract: Inside the Cell published by Wiley Periodicals, Inc.


Epidemiology of sepsis and septic shock in critical care units: comparison between sepsis-2 and sepsis-3 populations using a national critical care database

Abstract: Background New sepsis and septic shock definitions could change the epidemiology of sepsis because of differences in criteria.

Abstract: We therefore compared the sepsis populations identified by the old and new definitions.

Abstract: Methods We used a high-quality, national, intensive care unit (ICU) database of 654 918 consecutive admissions to 189 adult ICUs in England, from January 2011 to December 2015.

Abstract: Primary outcome was acute hospital mortality.

Abstract: We compared old (Sepsis-2) and new (Sepsis-3) incidence, outcomes, trends in outcomes, and predictive validity of sepsis and septic shock populations.

Abstract: Results From among 197 724 Sepsis-2 severe sepsis and 197 142 Sepsis-3 sepsis cases, we identified 153 257 Sepsis-2 septic shock and 39 262 Sepsis-3 septic shock cases.

Abstract: The extrapolated population incidence of Sepsis-3 sepsis and Sepsis-3 septic shock was 101.8 and 19.3 per 100 000 person-years, respectively, in 2015.

Abstract: Sepsis-2 severe sepsis and Sepsis-3 sepsis had similar incidence, similar mortality and showed significant risk-adjusted improvements in mortality over time.

Abstract: Sepsis-3 septic shock had a much higher Acute Physiology And Chronic Health Evaluation II (APACHE II) score, greater mortality and no risk-adjusted trends in mortality improvement compared with Sepsis-2 septic shock.

Abstract: ICU admissions identified either as Sepsis-3 sepsis or septic shock and as Sepsis-2 severe sepsis or septic shock had significantly greater risk-adjusted odds of death compared with non-sepsis admissions (P<0.001).

Abstract: The predictive validity was greatest for Sepsis-3 septic shock.

Abstract: Conclusions In an ICU database, compared with Sepsis-2, Sepsis-3 identifies a similar sepsis population with 92% overlap and much smaller septic shock population with improved predictive validity.


Thymic expression of the golli-myelin basic protein gene in the SJL/J mouse

Abstract: The T cell antigen-specific repertoire is thought to be shaped by thymic expression of self molecules.

Abstract: Since a myelin basic protein (MBP)-like gene (golli-MBP) has been reported to be expressed by cells of the immune system, the present study was undertaken to determine whether the golli-MBP gene was expressed in the mouse thymus and, if so, to characterize transcripts of this gene in this organ.

Abstract: Using exon-specific primers for MBP and golli-MBP, cDNA from thymus and other tissues was amplified, and the amplified products analyzed by Southern blotting with exon-specific oligonucleotide probes.

Abstract: The amplified products were subcloned, and the inserts characterized by DNA sequencing.

Abstract: The thymic transcripts were found to contain golli-MBP exons 1, 2, 3, 5A, 5B, 5C, 6, 7, 8, and 11.


Urine microRNAs as biomarkers for bladder cancer: a diagnostic meta-analysis

Abstract: BACKGROUND The diagnostic value of microRNA (miRNA) detection in patients with bladder cancer (BCa) is controversial.

Abstract: We performed a diagnostic meta-analysis to evaluate current evidence on the use of miRNA assays to diagnose BCa.

Abstract: METHODS We systematically searched PubMed, Embase, and Web of Science for studies published before March 31, 2015.

Abstract: The pooled sensitivity, specificity, positive and negative likelihood ratios, diagnostic odds ratio, and area under the curve (AUC) were calculated to evaluate the overall test performance.

Abstract: Subgroup analyses were used to explore the between-study heterogeneity.

Abstract: Deeks' funnel plot asymmetry test was used to test publication bias.

Abstract: We applied the software of RevMan 5.2 and Stata 11.0 to the meta-analysis.

Abstract: RESULTS A total of 23 studies from nine articles were included in the meta-analysis, with a total of 719 patients and 494 controls.

Abstract: The pooled sensitivity and specificity were 0.75 (95% confidence interval [CI], 0.68-0.80) and 0.75 (95% CI, 0.70-0.80), respectively.

Abstract: The pooled positive likelihood ratio was 3.03 (95% CI, 2.50-3.67); negative likelihood ratio was 0.33 (95% CI, 0.27-0.42); and diagnostic odds ratio was 9.07 (95% CI, 6.35-12.95).

Abstract: The pooled AUC was 0.81 (95% CI, 0.78-0.85).

Abstract: Subgroup analyses indicated that the multiple miRNAs assays and urine supernatant assays showed high accuracies in diagnosing BCa.

Abstract: CONCLUSION The miRNA assays may serve as potential noninvasive diagnostic tool for the detection of BCa.

Abstract: However, the clinical application of miRNA assays for BCa diagnosis still needs further validation by large prospective studies.


Elevated Plasma Transforming Growth Factor &bgr;1 Levels Predict the Development of Hypertension in Normotensives: The 14-Year Follow-Up Study

Abstract: BACKGROUND Transforming growth factor β1 (TGF-β1) is a multifunctional cytokine.

Abstract: There is growing evidence that TGF-β1 is involved in the pathogenesis of hypertension and the development of target organ damage in hypertensives.

Abstract: Although several studies have shown that TGF-β1 induced vascular hypertrophy and remodelling in various vascular diseases, there are no longitudinal data on hypertension in the epidemiological studies.

Abstract: The present study tested the hypothesis whether elevated TGF-β1 levels can predict the development of hypertension.

Abstract: METHODS In 2002-2004, 528 subjects received health examinations in Uku town, southwestern Japan.

Abstract: We examined blood pressure (BP), body mass index, and blood test.

Abstract: Data on fasting plasma TGF-β1 were obtained from 528 individuals.

Abstract: Of these, 149 normotensives (BP <140/90 mm Hg without antihypertensive medications) at baseline were followed-up for 14 years.

Abstract: RESULTS The receiver-operating characteristic curve was used and the calculated cutoff value was 8.9 ng/ml.

Abstract: Of 149 normotensives at baseline, 59 subjects developed hypertension.

Abstract: Plasma TGF-β1 levels were significantly associated with the development of hypertension after adjustment for confounding factors.

Abstract: To further examine the association between them, we performed logistic regression analysis.

Abstract: We divided the baseline plasma TGF-β1 levels into 2 groups using a cutoff value.

Abstract: The significant high odds ratio [3.582 (95% confidence interval, 1.025-12.525)] for the development of hypertension was found in the highest group of TGF-β1 level vs. the lowest group after adjustment for confounders.

Abstract: CONCLUSIONS This is the first report demonstrating the causal relationship between them.

Abstract: Elevated plasma TGF-β1 levels predicted the development of hypertension in normotensives in a population of community-dwelling Japanese.


MYC Disrupts the Circadian Clock and Metabolism in Cancer Cells.

Abstract: The MYC oncogene encodes MYC, a transcription factor that binds the genome through sites termed E-boxes (5'-CACGTG-3'), which are identical to the binding sites of the heterodimeric CLOCK-BMAL1 master circadian transcription factor.

Abstract: Hence, we hypothesized that ectopic MYC expression perturbs the clock by deregulating E-box-driven components of the circadian network in cancer cells.

Abstract: We report here that deregulated expression of MYC or N-MYC disrupts the molecular clock in vitro by directly inducing REV-ERBα to dampen expression and oscillation of BMAL1, and this could be rescued by knockdown of REV-ERB.

Abstract: REV-ERBα expression predicts poor clinical outcome for N-MYC-driven human neuroblastomas that have diminished BMAL1 expression, and re-expression of ectopic BMAL1 in neuroblastoma cell lines suppresses their clonogenicity.

Abstract: Further, ectopic MYC profoundly alters oscillation of glucose metabolism and perturbs glutaminolysis.

Abstract: Our results demonstrate an unsuspected link between oncogenic transformation and circadian and metabolic dysrhythmia, which we surmise to be advantageous for cancer.


Integrated maps in quail (Coturnix japonica) confirm the high degree of synteny conservation with chicken (Gallus gallus) despite 35 million years of divergence

Abstract: BackgroundBy comparing the quail genome with that of chicken, chromosome rearrangements that have occurred in these two galliform species over 35 million years of evolution can be detected.

Abstract: From a more practical point of view, the definition of conserved syntenies helps to predict the position of genes in quail, based on information taken from the chicken sequence, thus enhancing the utility of this species in biological studies through a better knowledge of its genome structure.

Abstract: A microsatellite and an Amplified Fragment Length Polymorphism (AFLP) genetic map were previously published for quail, as well as comparative cytogenetic data with chicken for macrochromosomes.

Abstract: Quail genomics will benefit from the extension and the integration of these maps.

Abstract: ResultsThe integrated linkage map presented here is based on segregation analysis of both anonymous markers and functional gene loci in 1,050 quail from three independent F2 populations.

Abstract: Ninety-two loci are resolved into 14 autosomal linkage groups and a Z chromosome-specific linkage group, aligned with the quail AFLP map.

Abstract: The size of linkage groups ranges from 7.8 cM to 274.8 cM. The total map distance covers 904.3 cM with an average spacing of 9.7 cM between loci.

Abstract: The coverage is not complete, as macrochromosome CJA08, the gonosome CJAW and 23 microchromosomes have no marker assigned yet.

Abstract: Significant sequence identities of quail markers with chicken enabled the alignment of the quail linkage groups on the chicken genome sequence assembly.

Abstract: This, together with interspecific Fluorescence In Situ Hybridization (FISH), revealed very high similarities in marker order between the two species for the eight macrochromosomes and the 14 microchromosomes studied.

Abstract: ConclusionIntegrating the two microsatellite and the AFLP quail genetic maps greatly enhances the quality of the resulting information and will thus facilitate the identification of Quantitative Trait Loci (QTL).

Abstract: The alignment with the chicken chromosomes confirms the high conservation of gene order that was expected between the two species for macrochromosomes.

Abstract: By extending the comparative study to the microchromosomes, we suggest that a wealth of information can be mined in chicken, to be used for genome analyses in quail.


Cadherin activity is required for activity-induced spine remodeling

Abstract: Neural activity induces the remodeling of pre- and postsynaptic membranes, which maintain their apposition through cell adhesion molecules.

Abstract: Among them, N-cadherin is redistributed, undergoes activity-dependent conformational changes, and is required for synaptic plasticity.

Abstract: Here, we show that depolarization induces the enlargement of the width of spine head, and that cadherin activity is essential for this synaptic rearrangement.

Abstract: Dendritic spines visualized with green fluorescent protein in hippocampal neurons showed an expansion by the activation of AMPA receptor, so that the synaptic apposition zone may be expanded.

Abstract: N-cadherin-venus fusion protein laterally dispersed along the expanding spine head.

Abstract: Overexpression of dominant-negative forms of N-cadherin resulted in the abrogation of the spine expansion.

Abstract: Inhibition of actin polymerization with cytochalasin D abolished the spine expansion.

Abstract: Together, our data suggest that cadherin-based adhesion machinery coupled with the actin-cytoskeleton is critical for the remodeling of synaptic apposition zone.


Aliskiren combined with losartan in type 2 diabetes and nephropathy.

Abstract: BACKGROUND Diabetic nephropathy is the leading cause of end-stage renal disease in developed countries.

Abstract: We evaluated the renoprotective effects of dual blockade of the renin-angiotensin-aldosterone system by adding treatment with aliskiren, an oral direct renin inhibitor, to treatment with the maximal recommended dose of losartan (100 mg daily) and optimal antihypertensive therapy in patients who had hypertension and type 2 diabetes with nephropathy.

Abstract: METHODS We enrolled 599 patients in this multinational, randomized, double-blind study.

Abstract: After a 3-month, open-label, run-in period during which patients received 100 mg of losartan daily, patients were randomly assigned to receive 6 months of treatment with aliskiren (150 mg daily for 3 months, followed by an increase in dosage to 300 mg daily for another 3 months) or placebo, in addition to losartan.

Abstract: The primary outcome was a reduction in the ratio of albumin to creatinine, as measured in an early-morning urine sample, at 6 months.

Abstract: RESULTS The baseline characteristics of the two groups were similar.

Abstract: Treatment with 300 mg of aliskiren daily, as compared with placebo, reduced the mean urinary albumin-to-creatinine ratio by 20% (95% confidence interval, 9 to 30; P<0.001), with a reduction of 50% or more in 24.7% of the patients who received aliskiren as compared with 12.5% of those who received placebo (P<0.001).

Abstract: A small difference in blood pressure was seen between the treatment groups by the end of the study period (systolic, 2 mm Hg lower [P=0.07] and diastolic, 1 mm Hg lower [P=0.08] in the aliskiren group).

Abstract: The total numbers of adverse and serious adverse events were similar in the groups.

Abstract: CONCLUSIONS Aliskiren may have renoprotective effects that are independent of its blood-pressure-lowering effect in patients with hypertension, type 2 diabetes, and nephropathy who are receiving the recommended renoprotective treatment.

Abstract: (ClinicalTrials.gov number, NCT00097955 [ClinicalTrials.gov].).


Overweight in Celiac Disease: Prevalence, Clinical Characteristics, and Effect of a Gluten-Free Diet

Abstract: BACKGROUND:It is well established that a minority of celiac patients present with “classic” symptoms due to malabsorption.

Abstract: However, few studies have focussed on the distribution of body mass index (BMI) in celiac populations and its relationship to clinical characteristics, or on its response to treatment.

Abstract: METHODS:We reviewed BMI measurements and other clinical and pathological characteristics from a database of 371 celiac patients diagnosed over a 10-yr period and seen by a single gastroenterologist.

Abstract: To assess response to gluten exclusion, we compared BMI at diagnosis and after 2 yr treatment in patients with serological support for dietary compliance.

Abstract: RESULTS:Mean BMI was 24.6 kg/m2 (range 16.3–43.5).

Abstract: Seventeen patients (5%) were underweight (BMI <18.5), 211 (57%) were normal, and 143 (39%) were overweight (BMI ≥25), including 48 (13% of all patients) in the obese range (BMI ≥30.0).

Abstract: There was a significant association between low BMI and female gender, history of diarrhea, reduced hemoglobin concentration, reduced bone mineral density (BMD), osteoporosis, and higher grades (subtotal/total) of villous atrophy.

Abstract: Of patients compliant with a gluten-free diet, 81% had gained weight after 2 yr, including 82% of initially overweight patients.

Abstract: CONCLUSIONS:Few celiac patients are underweight at diagnosis and a large minority is overweight; these are less likely to present with classical features of diarrhea and reduced hemoglobin.

Abstract: Failed or delayed diagnosis of celiac disease may reflect lack of awareness of this large subgroup.

Abstract: The increase in weight of already overweight patients after dietary gluten exclusion is a potential cause of morbidity, and the gluten-free diet as conventionally prescribed needs to be modified accordingly.


Structural Studies of a Four-MBT Repeat Protein MBTD1

Abstract: BACKGROUND The Polycomb group (PcG) of proteins is a family of important developmental regulators.

Abstract: The respective members function as large protein complexes involved in establishment and maintenance of transcriptional repression of developmental control genes.

Abstract: MBTD1, Malignant Brain Tumor domain-containing protein 1, is one such PcG protein.

Abstract: MBTD1 contains four MBT repeats.

Abstract: METHODOLOGY/PRINCIPAL FINDINGS We have determined the crystal structure of MBTD1 (residues 130-566aa covering the 4 MBT repeats) at 2.5 A resolution by X-ray crystallography.

Abstract: The crystal structure of MBTD1 reveals its similarity to another four-MBT-repeat protein L3MBTL2, which binds lower methylated lysine histones.

Abstract: Fluorescence polarization experiments confirmed that MBTD1 preferentially binds mono- and di-methyllysine histone peptides, like L3MBTL1 and L3MBTL2.

Abstract: All known MBT-peptide complex structures characterized to date do not exhibit strong histone peptide sequence selectivity, and use a "cavity insertion recognition mode" to recognize the methylated lysine with the deeply buried methyl-lysine forming extensive interactions with the protein while the peptide residues flanking methyl-lysine forming very few contacts [1].

Abstract: Nevertheless, our mutagenesis data based on L3MBTL1 suggested that the histone peptides could not bind to MBT repeats in any orientation.

Abstract: CONCLUSIONS The four MBT repeats in MBTD1 exhibits an asymmetric rhomboid architecture.

Abstract: Like other MBT repeat proteins characterized so far, MBTD1 binds mono- or dimethylated lysine histones through one of its four MBT repeats utilizing a semi-aromatic cage.

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Abstract: Instructions for the installation and use of the web plugin are available in Text S1.


Glutamine supports pancreatic cancer growth through a Kras-regulated metabolic pathway

Abstract: Cancer cells have metabolic dependencies that distinguish them from their normal counterparts.

Abstract: Among these dependencies is an increased use of the amino acid glutamine to fuel anabolic processes.

Abstract: Indeed, the spectrum of glutamine-dependent tumours and the mechanisms whereby glutamine supports cancer metabolism remain areas of active investigation.

Abstract: Here we report the identification of a non-canonical pathway of glutamine use in human pancreatic ductal adenocarcinoma (PDAC) cells that is required for tumour growth.

Abstract: Whereas most cells use glutamate dehydrogenase (GLUD1) to convert glutamine-derived glutamate into α-ketoglutarate in the mitochondria to fuel the tricarboxylic acid cycle, PDAC relies on a distinct pathway in which glutamine-derived aspartate is transported into the cytoplasm where it can be converted into oxaloacetate by aspartate transaminase (GOT1).

Abstract: Subsequently, this oxaloacetate is converted into malate and then pyruvate, ostensibly increasing the NADPH/NADP(+) ratio which can potentially maintain the cellular redox state.

Abstract: Importantly, PDAC cells are strongly dependent on this series of reactions, as glutamine deprivation or genetic inhibition of any enzyme in this pathway leads to an increase in reactive oxygen species and a reduction in reduced glutathione.

Abstract: Moreover, knockdown of any component enzyme in this series of reactions also results in a pronounced suppression of PDAC growth in vitro and in vivo.

Abstract: Furthermore, we establish that the reprogramming of glutamine metabolism is mediated by oncogenic KRAS, the signature genetic alteration in PDAC, through the transcriptional upregulation and repression of key metabolic enzymes in this pathway.

Abstract: The essentiality of this pathway in PDAC and the fact that it is dispensable in normal cells may provide novel therapeutic approaches to treat these refractory tumours.


CD4+ T cell help guides formation of CD103+ lung-resident memory CD8+ T cells during influenza viral infection.

Abstract: Tissue-resident memory T (Trm) cells provide enhanced protection against infection at mucosal sites.

Abstract: Here we found that CD4(+) T cells are important for the formation of functional lung-resident CD8(+) T cells after influenza virus infection.

Abstract: In the absence of CD4(+) T cells, CD8(+) T cells displayed reduced expression of CD103 (Itgae), were mislocalized away from airway epithelia, and demonstrated an impaired ability to recruit CD8(+) T cells to the lung airways upon heterosubtypic challenge.

Abstract: CD4(+) T cell-derived interferon-γ was necessary for generating lung-resident CD103(+) CD8(+) Trm cells.

Abstract: Furthermore, expression of the transcription factor T-bet was increased in "unhelped" lung Trm cells, and a reduction in T-bet rescued CD103 expression in the absence of CD4(+) T cell help.

Abstract: Thus, CD4(+) T cell-dependent signals are important to limit expression of T-bet and allow for the development of CD103(+) CD8(+) Trm cells in the lung airways following respiratory infection.


Inhibition of Apoptosis Overcomes Stage-Related Compatibility Barriers to Chimera Formation in Mouse Embryos.

Abstract: Cell types more advanced in development than embryonic stem cells, such as EpiSCs, fail to contribute to chimeras when injected into pre-implantation-stage blastocysts, apparently because the injected cells undergo apoptosis.

Abstract: Here we show that transient promotion of cell survival through expression of the anti-apoptotic gene BCL2 enables EpiSCs and Sox17+ endoderm progenitors to integrate into blastocysts and contribute to chimeric embryos.

Abstract: Upon injection into blastocyst, BCL2-expressing EpiSCs contributed to all bodily tissues in chimeric animals while Sox17+ endoderm progenitors specifically contributed in a region-specific fashion to endodermal tissues.

Abstract: In addition, BCL2 expression enabled rat EpiSCs to contribute to mouse embryonic chimeras, thereby forming interspecies chimeras that could survive to adulthood.

Abstract: Our system therefore provides a method to overcome cellular compatibility issues that typically restrict chimera formation.

Abstract: Application of this type of approach could broaden the use of embryonic chimeras, including region-specific chimeras, for basic developmental biology research and regenerative medicine.


Computational and Statistical Analyses of Amino Acid Usage and Physico-Chemical Properties of the Twelve Late Embryogenesis Abundant Protein Classes

Abstract: Late Embryogenesis Abundant Proteins (LEAPs) are ubiquitous proteins expected to play major roles in desiccation tolerance.

Abstract: Little is known about their structure - function relationships because of the scarcity of 3-D structures for LEAPs.

Abstract: The previous building of LEAPdb, a database dedicated to LEAPs from plants and other organisms, led to the classification of 710 LEAPs into 12 non-overlapping classes with distinct properties.

Abstract: Using this resource, numerous physico-chemical properties of LEAPs and amino acid usage by LEAPs have been computed and statistically analyzed, revealing distinctive features for each class.

Abstract: This unprecedented analysis allowed a rigorous characterization of the 12 LEAP classes, which differed also in multiple structural and physico-chemical features.

Abstract: Although most LEAPs can be predicted as intrinsically disordered proteins, the analysis indicates that LEAP class 7 (PF03168) and probably LEAP class 11 (PF04927) are natively folded proteins.

Abstract: This study thus provides a detailed description of the structural properties of this protein family opening the path toward further LEAP structure - function analysis.

Abstract: Finally, since each LEAP class can be clearly characterized by a unique set of physico-chemical properties, this will allow development of software to predict proteins as LEAPs.


Direct Reprogramming of Hepatic Myofibroblasts into Hepatocytes In Vivo Attenuates Liver Fibrosis.

Abstract: Direct induction of induced hepatocytes (iHeps) from fibroblasts holds potential as a strategy for regenerative medicine but until now has only been shown in culture settings.

Abstract: Here, we describe in vivo iHep formation using transcription factor induction and genetic fate tracing in mouse models of chronic liver disease.

Abstract: We show that ectopic expression of the transcription factors FOXA3, GATA4, HNF1A, and HNF4A from a polycistronic lentiviral vector converts mouse myofibroblasts into cells with a hepatocyte phenotype.

Abstract: In vivo expression of the same set of transcription factors from a p75 neurotrophin receptor peptide (p75NTRp)-tagged adenovirus enabled the generation of hepatocyte-like cells from myofibroblasts in fibrotic mouse livers and reduced liver fibrosis.

Abstract: We have therefore been able to convert pro-fibrogenic myofibroblasts in the liver into hepatocyte-like cells with positive functional benefits.

Abstract: This direct in vivo reprogramming approach may open new avenues for the treatment of chronic liver disease.


Epithelial to Mesenchymal Transition by TGFβ-1 Induction Increases Stemness Characteristics in Primary Non Small Cell Lung Cancer Cell Line

Abstract: BACKGROUND Cancer Stem Cells (CSCs) hypothesis asserts that only a small subset of cells within a tumour is capable of both tumour initiation and sustainment.

Abstract: The Epithelial-Mesenchymal Transition (EMT) is an embryonic developmental program that is often activated during cancer invasion and metastasis.

Abstract: The aim of this study is to shed light on the relationship between EMT and CSCs by using LC31 lung cancer primary cell line.

Abstract: MATERIALS AND METHODS A549 and LC31 cell lines were treated with 2 ng/ml TGFβ-1 for 30 days, and 80 days, respectively.

Abstract: To evaluate EMT, morphological changes were assessed by light microscopy, immunofluorescence and cytometry for following markers: cytokeratins, e-cadherin, CD326 (epithelial markers) and CD90, and vimentin (mesenchymal markers).

Abstract: Moreover, RT-PCR for Slug, Twist and β-catenin genes were performed.

Abstract: On TGFβ-1 treated and untreated LC31 cell lines, we performed stemness tests such as pneumospheres growth and stem markers expression such as Oct4, Nanog, Sox2, c-kit and CD133.

Abstract: Western Blot for CD133 and tumorigenicity assays using NOD/SCID mice were performed.

Abstract: RESULTS TGFβ-1 treated LC31 cell line lost its epithelial morphology assuming a fibroblast-like appearance.

Abstract: The same results were obtained for the A549 cell line (as control).

Abstract: Immunofluorescence and cytometry showed up-regulation of vimentin and CD90 and down-regulation of cytocheratin, e-cadherin and CD326 in TGFβ-1 treated LC31 and A549 cell lines.

Abstract: Slug, Twist and β-catenin m-RNA transcripts were up-regulated in TGFβ-1 treated LC31 cell line confirming EMT.

Abstract: This cell line showed also over-expression of Oct4, Nanog, Sox2 and CD133, all genes of stemness.

Abstract: In addition, in TGFβ-1 treated LC31 cell line, an increased pneumosphere-forming capacity and tumours-forming ability in NOD/SCID mice were detectable.

Abstract: CONCLUSIONS The induction of EMT by TGFβ-1 exposure, in primary lung cancer cell line results in the acquisition of mesenchymal profile and in the expression of stem cell markers.


Dnmt3a and Dnmt3b Associate with Enhancers to Regulate Human Epidermal Stem Cell Homeostasis.

Abstract: The genome-wide localization and function of endogenous Dnmt3a and Dnmt3b in adult stem cells are unknown.

Abstract: Here, we show that in human epidermal stem cells, the two proteins bind in a histone H3K36me3-dependent manner to the most active enhancers and are required to produce their associated enhancer RNAs.

Abstract: Both proteins prefer super-enhancers associated to genes that either define the ectodermal lineage or establish the stem cell and differentiated states.

Abstract: However, Dnmt3a and Dnmt3b differ in their mechanisms of enhancer regulation: Dnmt3a associates with p63 to maintain high levels of DNA hydroxymethylation at the center of enhancers in a Tet2-dependent manner, whereas Dnmt3b promotes DNA methylation along the body of the enhancer.

Abstract: Depletion of either protein inactivates their target enhancers and profoundly affects epidermal stem cell function.

Abstract: Altogether, we reveal novel functions for Dnmt3a and Dnmt3b at enhancers that could contribute to their roles in disease and tumorigenesis.


CDK inhibitors in cancer therapy: what is next?

Abstract: The pursuit for drugs that inhibit cyclin-dependent kinases (CDKs) has been an intense area of research for more than 15 years.

Abstract: The first-generation inhibitors, Flavopiridol and CY-202, are in late-stage clinical trials, but so far have demonstrated only modest activity.

Abstract: Several second-generation inhibitors are now in clinical trials.

Abstract: Future approaches to determine clinical benefit need to incorporate both the lessons learned from these early compounds and information recently obtained from the genetic analysis of CDKs in preclinical models.

Abstract: Here we discuss key concepts that should be considered when validating the clinical utility of CDK inhibitors in cancer therapy.


Lipid Desaturation Is a Metabolic Marker and Therapeutic Target of Ovarian Cancer Stem Cells.

Abstract: Lack of sensitive single-cell analysis tools has limited the characterization of metabolic activity in cancer stem cells.

Abstract: By hyperspectral-stimulated Raman scattering imaging of single living cells and mass spectrometry analysis of extracted lipids, we report here significantly increased levels of unsaturated lipids in ovarian cancer stem cells (CSCs) as compared to non-CSCs.

Abstract: Higher lipid unsaturation levels were also detected in CSC-enriched spheroids compared to monolayer cultures of ovarian cancer cell lines or primary cells.

Abstract: Inhibition of lipid desaturases effectively eliminated CSCs, suppressed sphere formation in vitro, and blocked tumor initiation capacity in vivo.

Abstract: Mechanistically, we demonstrate that nuclear factor κB (NF-κB) directly regulates the expression levels of lipid desaturases, and inhibition of desaturases blocks NF-κB signaling.

Abstract: Collectively, our findings reveal that increased lipid unsaturation is a metabolic marker for ovarian CSCs and a target for CSC-specific therapy.


Mesenchymal Inflammation Drives Genotoxic Stress in Hematopoietic Stem Cells and Predicts Disease Evolution in Human Pre-leukemia.

Abstract: Mesenchymal niche cells may drive tissue failure and malignant transformation in the hematopoietic system, but the underlying molecular mechanisms and relevance to human disease remain poorly defined.

Abstract: Here, we show that perturbation of mesenchymal cells in a mouse model of the pre-leukemic disorder Shwachman-Diamond syndrome (SDS) induces mitochondrial dysfunction, oxidative stress, and activation of DNA damage responses in hematopoietic stem and progenitor cells.

Abstract: Massive parallel RNA sequencing of highly purified mesenchymal cells in the SDS mouse model and a range of human pre-leukemic syndromes identified p53-S100A8/9-TLR inflammatory signaling as a common driving mechanism of genotoxic stress.

Abstract: Transcriptional activation of this signaling axis in the mesenchymal niche predicted leukemic evolution and progression-free survival in myelodysplastic syndrome (MDS), the principal leukemia predisposition syndrome.

Abstract: Collectively, our findings identify mesenchymal niche-induced genotoxic stress in heterotypic stem and progenitor cells through inflammatory signaling as a targetable determinant of disease outcome in human pre-leukemia.


Novel Functional Sets of Lipid-Derived Mediators with Antiinflammatory Actions Generated from Omega-3 Fatty Acids via Cyclooxygenase 2–Nonsteroidal Antiinflammatory Drugs and Transcellular Processing

Abstract: Aspirin therapy inhibits prostaglandin biosynthesis without directly acting on lipoxygenases, yet via acetylation of cyclooxygenase 2 (COX-2) it leads to bioactive lipoxins (LXs) epimeric at carbon 15 (15-epi-LX, also termed aspirin-triggered LX [ATL]).

Abstract: Here, we report that inflammatory exudates from mice treated with ω-3 polyunsaturated fatty acid and aspirin (ASA) generate a novel array of bioactive lipid signals.

Abstract: Human endothelial cells with upregulated COX-2 treated with ASA converted C20:5 ω-3 to 18R-hydroxyeicosapentaenoic acid (HEPE) and 15R-HEPE.

Abstract: Each was used by polymorphonuclear leukocytes to generate separate classes of novel trihydroxy-containing mediators, including 5-series 15R-LX5 and 5,12,18R-triHEPE.

Abstract: These new compounds proved to be potent inhibitors of human polymorphonuclear leukocyte transendothelial migration and infiltration in vivo (ATL analogue > 5,12,18R-triHEPE > 18R-HEPE).

Abstract: Acetaminophen and indomethacin also permitted 18R-HEPE and 15R-HEPE generation with recombinant COX-2 as well as ω-5 and ω-9 oxygenations of other fatty acids that act on hematologic cells.

Abstract: These findings establish new transcellular routes for producing arrays of bioactive lipid mediators via COX-2–nonsteroidal antiinflammatory drug–dependent oxygenations and cell–cell interactions that impact microinflammation.

Abstract: The generation of these and related compounds provides a novel mechanism(s) for the therapeutic benefits of ω-3 dietary supplementation, which may be important in inflammation, neoplasia, and vascular diseases.


Cholesteryl Ester Transfer Protein (CETP) Polymorphisms Affect mRNA Splicing, HDL Levels, and Sex-Dependent Cardiovascular Risk

Abstract: Polymorphisms in and around the Cholesteryl Ester Transfer Protein (CETP) gene have been associated with HDL levels, risk for coronary artery disease (CAD), and response to therapy.

Abstract: The mechanism of action of these polymorphisms has yet to be defined.

Abstract: We used mRNA allelic expression and splice isoform measurements in human liver tissues to identify the genetic variants affecting CETP levels.

Abstract: Allelic CETP mRNA expression ratios in 56 human livers were strongly associated with several variants 2.5-7 kb upstream of the transcription start site (e.g., rs247616 p = 6.4 × 10(-5), allele frequency 33%).

Abstract: In addition, a common alternatively spliced CETP isoform lacking exon 9 (Δ9), has been shown to prevent CETP secretion in a dominant-negative manner.

Abstract: The Δ 9 expression ranged from 10 to 48% of total CETP mRNA in 94 livers.

Abstract: Increased formation of this isoform was exclusively associated with an exon 9 polymorphism rs5883-C>T (p = 6.8 × 10(-10)) and intron 8 polymorphism rs9930761-T>C (5.6 × 10(-8)) (in high linkage disequilibrium with allele frequencies 6-7%).

Abstract: rs9930761 changes a key splicing branch point nucleotide in intron 8, while rs5883 alters an exonic splicing enhancer sequence in exon 9.The effect of these polymorphisms was evaluated in two clinical studies.

Abstract: In the Whitehall II study of 4745 subjects, both rs247616 and rs5883T/rs9930761C were independently associated with increased HDL-C levels in males with similar effect size (rs247616 p = 9.6 × 10(-28) and rs5883 p = 8.6 × 10(-10), adjusted for rs247616).

Abstract: In an independent multiethnic US cohort of hypertensive subjects with CAD (INVEST-GENE), rs5883T/rs9930761C alone were significantly associated with increased incidence of MI, stroke, and all-cause mortality in males (rs5883: OR 2.36 (CI 1.29-4.30), p = 0.005, n = 866).

Abstract: These variants did not reach significance in females in either study.

Abstract: Similar to earlier results linking low CETP activity with poor outcomes in males, our results suggest genetic, sex-dependent CETP splicing effects on cardiovascular risk by a mechanism independent of circulating HDL-C levels.