Microtubule sliding activity of a kinesin-8 promotes spindle assembly and spindle length control

Abstract: Molecular motors play critical roles in the formation of mitotic spindles, either through controlling the stability of individual microtubules, or by crosslinking and sliding microtubule arrays.

Abstract: Kinesin-8 motors are best known for their regulatory roles in controlling microtubule dynamics.

Abstract: They contain microtubule-destabilizing activities, and restrict spindle length in a wide variety of cell types and organisms.

Abstract: Here, we report an antiparallel microtubule-sliding activity of the budding yeast kinesin-8, Kip3.

Abstract: The in vivo importance of this sliding activity was established through the identification of complementary Kip3 mutants that separate the sliding activity and microtubule-destabilizing activity.

Abstract: In conjunction with Cin8, a kinesin-5 family member, the sliding activity of Kip3 promotes bipolar spindle assembly and the maintenance of genome stability.

Abstract: We propose a slide-disassemble model where the sliding and destabilizing activity of Kip3 balance during pre-anaphase.

Abstract: This facilitates normal spindle assembly.

Abstract: However, the destabilizing activity of Kip3 dominates in late anaphase, inhibiting spindle elongation and ultimately promoting spindle disassembly.


A Pharmacological Map of the PI3-K Family Defines a Role for p110α in Insulin Signaling

Abstract: Phosphoinositide 3-kinases (PI3-Ks) are an important emerging class of drug targets, but the unique roles of PI3-K isoforms remain poorly defined.

Abstract: We describe here an approach to pharmacologically interrogate the PI3-K family.

Abstract: A chemically diverse panel of PI3-K inhibitors was synthesized, and their target selectivity was biochemically enumerated, revealing cryptic homologies across targets and chemotypes.

Abstract: Crystal structures of three inhibitors bound to p110gamma identify a conformationally mobile region that is uniquely exploited by selective compounds.

Abstract: This chemical array was then used to define the PI3-K isoforms required for insulin signaling.

Abstract: We find that p110alpha is the primary insulin-responsive PI3-K in cultured cells, whereas p110beta is dispensable but sets a phenotypic threshold for p110alpha activity.

Abstract: Compounds targeting p110alpha block the acute effects of insulin treatment in vivo, whereas a p110beta inhibitor has no effect.

Abstract: These results illustrate systematic target validation using a matrix of inhibitors that span a protein family.


Identification of Epstein-Barr virus nuclear antigen 1 protein domains that direct interactions at a distance between DNA-bound proteins.

Abstract: The EBNA1 protein of Epstein-Barr virus (EBV) binds to and activates DNA replication from the EBV latent origin of replication, oriP, via a direct interaction with the two noncontiguous subelements of oriP. The EBNA1 molecules bound to the oriP subelements interact efficiently with each other by a DNA looping mechanism.

Abstract: We have previously mapped a region of EBNA1 (termed the looping region) that is required to mediate the interaction of the EBNA1 molecules bound to the oriP subelements.

Abstract: We now demonstrate that two fragments of this region of EBNA1, which consist largely of an eight amino acid repeat, can mediate homotypic interactions when transferred to another DNA-binding protein.

Abstract: Protein interactions mediated by the EBNA1 looping region appear to be dependent on DNA binding since these interactions were detected between DNA-bound forms of the proteins only.


Primary prevention of hypertension: clinical and public health advisory from The National High Blood Pressure Education Program.

Abstract: The National High Blood Pressure Education Program Coordinating Committee published its first statement on the primary prevention of hypertension in 1993.

Abstract: This article updates the 1993 report, using new and further evidence from the scientific literature.

Abstract: Current recommendations for primary prevention of hypertension involve a population-based approach and an intensive targeted strategy focused on individuals at high risk for hypertension.

Abstract: These 2 strategies are complementary and emphasize 6 approaches with proven efficacy for prevention of hypertension: engage in moderate physical activity; maintain normal body weight; limit alcohol consumption; reduce sodium intake; maintain adequate intake of potassium; and consume a diet rich in fruits, vegetables, and low-fat dairy products and reduced in saturated and total fat.

Abstract: Applying these approaches to the general population as a component of public health and clinical practice can help prevent blood pressure from increasing and can help decrease elevated blood pressure levels for those with high normal blood pressure or hypertension.


Formins: intermediates in signal-transduction cascades that affect cytoskeletal reorganization.

Abstract: The control of cell growth and polarity depends on a dynamic actin cytoskeleton that has the ability to reorganize in response to developmental and environmental stimuli.

Abstract: In animals and fungi, formins are just one of the four major classes of poly-L-proline-containing (PLP) proteins that form part of the signal-transduction cascade that leads to rearrangement of the actin cytoskeleton.

Abstract: Analysis of the Arabidopsis genome sequence indicates that, unlike animals and fungi, formins are the only class of conserved profilin-binding PLP proteins in plants.

Abstract: Moreover, plant formins show significant structural differences compared with their animal and fungal counterparts, raising the possibility that plant formins are subject to novel mechanisms of control or perform unique roles in plants.


Genetic variability in GLP-1 receptor is associated with inter-individual differences in weight lowering potential of liraglutide in obese women with PCOS: a pilot study

Abstract: The weight lowering potential of glucagon-like peptide 1 (GLP-1) receptor agonists (RAs) is inter-individually different and clinically unpredictable.

Abstract: The potential role of genetic variability of GLP-1R on body weight response to GLP-1 RAs in obese women with polycystic ovary syndrome (PCOS) has not yet been evaluated.

Abstract: Fifty-seven obese women with PCOS (aged 30.7 ± 7.0, BMI 38.6 ± 5.3 kg/m2) were assigned to liraglutide 1.2 mg QD s.c.

Abstract: for 12 weeks and classified as strong responders regarding weight loss if they lost 5 % or more of their initial body weight.

Abstract: They were genotyped for common GLP-1R single nucleotide polymorphisms (SNPs) rs6923761 and rs10305420.

Abstract: Changes of measures of obesity were measured before and at the end of the treatment.

Abstract: Twenty out of 57 subjects were strong responders and lost 7.38 ± 1.74 compared to 2.11 ± 2.17 kg lost in poor responders.

Abstract: Carriers of at least one polymorphic rs10305420 allele had poor treatment response compared to carriers of two wild type alleles (OR = 0.27, 95 % CI = 0.09–0.85, P = 0.025).

Abstract: Carriers of at least one polymorphic rs6923761 allele tended to have stronger treatment response compared to carriers of two wild type alleles (OR = 3.06, 95 % CI = 0.96–9.74, P = 0.058).

Abstract: Fasting glucose and glucose after oral glucose tolerance test (OGTT) comparably decreased in both groups when compared to baseline, whereas no within treatment differences were found in androgen profile.

Abstract: Gastrointestinal adverse events were transit and balanced between strong and poor responders.

Abstract: GLP-1R rs10305420 polymorphism explained some of the inter-individual differences in response to liraglutide regarding weight loss in obese PCOS women.


Cost-effectiveness of aromatase inhibitor co-treatment for controlled ovarian stimulation.

Abstract: BACKGROUND To compare the clinical results and the cost-effectiveness of using the aromatase inhibitor, letrozole, in conjunction with FSH and FSH alone for controlled ovarian stimulation (COS) in patients undergoing intrauterine insemination (IUI) for a variety of indications.

Abstract: METHODS Four hundred and thirty-two consecutive patients who underwent 872 IUI cycles were included.

Abstract: The study population was composed of two groups.

Abstract: Group I included 308 patients who underwent 589 IUI cycles with letrozole and FSH for the following indications: anovulation (143 cycles), male factor infertility (147 cycles), unexplained infertility (250 cycles), endometriosis (18 cycles) and combined indications (31 cycles).

Abstract: Group II included 124 patients who underwent 283 IUI cycles who received FSH only for the following indications: ovarian factor infertility (82 cycles), male factor infertility (66 cycles), unexplained infertility (114 cycles), endometriosis (13 cycles) and other indications (8 cycles).

Abstract: Main outcome measures included number of mature follicles >16 mm in diameter, dose of FSH used per cycle, clinical pregnancy rate and cost-effectiveness ratio per pregnancy.

Abstract: RESULTS FSH dose required for ovarian stimulation was significantly lower when letrozole was used (P < 0.0001).

Abstract: Although a significantly higher number of follicles >16 mm and endometrial thickness at the day of hCG administration (P < 0.0001) were observed in Group II, pregnancy rate per started (14.4 versus 15.9%) and per completed cycles (15.77 versus 18.07%) was the same in Group I and Group II, respectively.

Abstract: IUI cancellation rate was significantly lower with letrozole treatment (P = 0.05%).

Abstract: The cost per cycle was significantly lower in Group I versus Group II (468.93 Can dollars +/-

Abstract: 418.18 versus 1067.28 +/-

Abstract: 921.43; P < 0.0001).

Abstract: The cost-effectiveness ratio was 3249.42 dollars in the letrozole group and 6712.00 dollars in the FSH-only group.

Abstract: CONCLUSION A letrozole-FSH combination could be an effective ovarian stimulation protocol in IUI cycles.

Abstract: Such a protocol may be more cost-effective than FSH alone because of the difference of FSH dose and cost.

Abstract: A randomized controlled trial is needed to further substantiate this finding.


The cost effectiveness of stereotactic radiosurgery versus surgical resection in the treatment of solitary metastatic brain tumors.

Abstract: Solitary metastatic brain tumors are the most common intracranial neoplasms encountered by neurosurgeons.

Abstract: Surgical resection of brain metastasis with whole brain radiotherapy (WBR) significantly increases survival in comparison with WBR alone.

Abstract: Stereotactic radiosurgery (SR) seems to provide results that are similar to those of surgical resection.

Abstract: To analyze the economic efficiency of these different treatments, we compared the results of surgical resection and SR as reported in the medical literature between 1974 and 1994.

Abstract: We included studies in which: 1) at least 75% of patients received WBR; 2) study dates were in the computed tomography era (after 1975); 3) operative morbidity, mortality, and median survival were reported; 4) study dates were not included in a more recent update or review; 5) tumor histologies were reported; and 6) the cobalt-60 gamma unit was used for SR.

Abstract: Three surgical resection studies and one SR study met all entry requirements.

Abstract: The WBR baseline was developed from two prospective, randomized trials and used for incremental cost effectiveness analysis.

Abstract: We developed a model of typical resource usage for uncomplicated procedures, reported complications, and subsequent craniotomies (for recurrent tumor or radiation necrosis) for both treatment options.

Abstract: Costs were estimated from the societal viewpoint using the 1992 Medicare Provider Analysis and Review database with average cost:charge ratios for surgery and WBR.

Abstract: A survey of capital and operating costs from five sites was used for radiosurgery.

Abstract: Our analysis revealed that radiosurgery had a lower uncomplicated procedure cost ($20,209 versus $27,587), a lower average complication cost per case ($2,534 versus $2,874), and a lower total cost per procedure ($22,743 versus $30,461), was more cost effective ($24,811 versus $32,149 per life year), and had a better incremental cost effectiveness ($40,648 versus $52,384 per life year) than surgical resection.

Abstract: A sensitivity analysis revealed that large changes in key assumptions would be required to change the analysis outcome.

Abstract: Equalization of the incremental cost effectiveness of the two treatments would require one of the following: 1) a 38.7% reduction in SR annual case volume, 2) a 34.7% increase in SR procedure cost, 3) a 18.8% reduction in surgical resection procedure cost, 4) a 240.5% increase in SR morbidity cost, 5) a 12.7% reduction in SR median survival, 6) a 16.8% increase in surgical resection median survival.

Abstract: Elimination of all surgical resection morbidity cost would still result in superior incremental cost effectiveness for SR.(ABSTRACT TRUNCATED AT 400 WORDS)


Trans-ancestry mutational landscape of hepatocellular carcinoma genomes

Abstract: Diverse epidemiological factors are associated with hepatocellular carcinoma (HCC) prevalence in different populations.

Abstract: However, the global landscape of the genetic changes in HCC genomes underpinning different epidemiological and ancestral backgrounds still remains uncharted.

Abstract: Here a collection of data from 503 liver cancer genomes from different populations uncovered 30 candidate driver genes and 11 core pathway modules.

Abstract: Furthermore, a collaboration of two large-scale cancer genome projects comparatively analyzed the trans-ancestry substitution signatures in 608 liver cancer cases and identified unique mutational signatures that predominantly contribute to Asian cases.

Abstract: This work elucidates previously unexplored ancestry-associated mutational processes in HCC development.

Abstract: A combination of hotspot TERT promoter mutation, TERT focal amplification and viral genome integration occurs in more than 68% of cases, implicating TERT as a central and ancestry-independent node of hepatocarcinogenesis.

Abstract: Newly identified alterations in genes encoding metabolic enzymes, chromatin remodelers and a high proportion of mTOR pathway activations offer potential therapeutic and diagnostic opportunities.


Organization of the mitotic chromosome.

Abstract: Mitotic chromosomes are among the most recognizable structures in the cell, yet for over a century their internal organization remains largely unsolved.

Abstract: We applied chromosome conformation capture methods, 5C and Hi-C, across the cell cycle and revealed two distinct three-dimensional folding states of the human genome.

Abstract: We show that the highly compartmentalized and cell type-specific organization described previously for nonsynchronous cells is restricted to interphase.

Abstract: In metaphase, we identified a homogenous folding state that is locus-independent, common to all chromosomes, and consistent among cell types, suggesting a general principle of metaphase chromosome organization.

Abstract: Using polymer simulations, we found that metaphase Hi-C data are inconsistent with classic hierarchical models and are instead best described by a linearly organized longitudinally compressed array of consecutive chromatin loops.


Compensatory anti-inflammatory response syndrome.

Abstract: The concept of 'Compensatory anti-inflammatory response syndrome' (CARS) was proposed in 1997 by Roger Bone (1941-1997) to qualify the consequences of the counter-regulatory mechanisms initiated to limit the overzealous inflammatory process in patients with infectious (sepsis) or non-infectious systemic inflammatory response syndrome (SIRS).

Abstract: One major consequence of CARS is the modification of the immune status that could favour the enhanced susceptibility of intensive care patients to nosocomial infections.

Abstract: Indeed, most animal 'two-hit' models illustrate an enhanced sensitivity to infection after a first insult.

Abstract: However, this observation is highly dependent on the experimental procedure.

Abstract: Numerous functions of circulating leukocytes are altered in sepsis and SIRS patients, as well as in animal models of sepsis or SIRS.

Abstract: However, this is rather a reprogramming of circulating leukocytes, since there is not a global defect of the immune cells functions.

Abstract: Furthermore, within tissues, leukocytes are rather primed or activated than immunosuppressed.

Abstract: Thus, CARS may be considered as an adapted compartmentalized response with the aim to silence some acute proinflammatory genes, and to maintain the possible expression of certain genes involved in the anti-infectious process.


Quantum dots spectrally distinguish multiple species within the tumor milieu in vivo

Abstract: A solid tumor is an organ composed of cancer and host cells embedded in an extracellular matrix and nourished by blood vessels.

Abstract: A prerequisite to understanding tumor pathophysiology is the ability to distinguish and monitor each component in dynamic studies.

Abstract: Standard fluorophores hamper simultaneous intravital imaging of these components.

Abstract: Here, we used multiphoton microscopy techniques and transgenic mice that expressed green fluorescent protein, and combined them with the use of quantum dot preparations.

Abstract: We show that these fluorescent semiconductor nanocrystals can be customized to concurrently image and differentiate tumor vessels from both the perivascular cells and the matrix.

Abstract: Moreover, we used them to measure the ability of particles of different sizes to access the tumor.

Abstract: Finally, we successfully monitored the recruitment of quantum dot–labeled bone marrow–derived precursor cells to the tumor vasculature.

Abstract: These examples show the versatility of quantum dots for studying tumor pathophysiology and creating avenues for treatment.


Arginase I: a limiting factor for nitric oxide and polyamine synthesis by activated macrophages?

Abstract: Because arginase hydrolyzes arginine to produce ornithine and urea, it has the potential to regulate nitric oxide (NO) and polyamine synthesis.

Abstract: We tested whether expression of the cytosolic isoform of arginase (arginase I) was limiting for NO or polyamine production by activated RAW 264.7 macrophage cells.

Abstract: RAW 264.7 cells, stably transfected to overexpress arginase I or beta-galactosidase, were treated with interferon-gamma to induce type 2 NO synthase or with lipopolysaccharide or 8-bromo-cAMP (8-BrcAMP) to induce ornithine decarboxylase.

Abstract: Overexpression of arginase I had no effect on NO synthesis.

Abstract: In contrast, cells overexpressing arginase I produced twice as much putrescine after activation than did cells expressing beta-galactosidase.

Abstract: Cells overexpressing arginase I also produced more spermidine after treatment with 8-BrcAMP than did cells expressing beta-galactosidase.

Abstract: Thus endogenous levels of arginase I are limiting for polyamine synthesis, but not for NO synthesis, by activated macrophage cells.

Abstract: This study also demonstrates that it is possible to alter arginase I levels sufficiently to affect polyamine synthesis without affecting induced NO synthesis.


Opposing Effects of PKCθ and WASp on Symmetry Breaking and Relocation of the Immunological Synapse

Abstract: The immunological synapse (IS) is a junction between the T cell and antigen-presenting cell and is composed of supramolecular activation clusters (SMACs).

Abstract: No studies have been published on naive T cell IS dynamics.

Abstract: Here, we find that IS formation during antigen recognition comprises cycles of stable IS formation and autonomous naive T cell migration.

Abstract: The migration phase is driven by PKCtheta, which is localized to the F-actin-dependent peripheral (p)SMAC.

Abstract: PKCtheta(-/-) T cells formed hyperstable IS in vitro and in vivo and, like WT cells, displayed fast oscillations in the distal SMAC, but they showed reduced slow oscillations in pSMAC integrity.

Abstract: IS reformation is driven by the Wiscott Aldrich Syndrome protein (WASp).

Abstract: WASp(-/-) T cells displayed normal IS formation but were unable to reform IS after migration unless PKCtheta was inhibited.

Abstract: Thus, opposing effects of PKCtheta and WASp control IS stability through pSMAC symmetry breaking and reformation.


Integrins regulate microtubule nucleating activity of centrosome through mitogen-activated protein kinase/extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase (MEK/ERK) signaling.

Abstract: Microtubule nucleation is an essential step in the formation of the microtubule cytoskeleton.

Abstract: We recently showed that androgen and Src promote microtubule nucleation and γ-tubulin accumulation at the centrosome.

Abstract: Here, we explore the mechanisms by which androgen and Src regulate these processes and ask whether integrins play a role.

Abstract: We perturb integrin function by a tyrosine-to-alanine substitution in membrane-proximal NPIY motif in the integrin β1 tail and show that this mutant substantially decreases microtubule nucleation and γ-tubulin accumulation at the centrosome.

Abstract: Because androgen stimulation promotes the interaction of the androgen receptor with Src, resulting in PI3K/AKT and MEK/ERK signaling, we asked whether these pathways are inhibited by the mutant integrin and whether they regulate microtubule nucleation.

Abstract: Our results indicate that the formation of the androgen receptor-Src complex and the activation of downstream pathways are significantly suppressed when cells are adhered by the mutant integrin.

Abstract: Inhibitor studies indicate that microtubule nucleation requires MEK/ERK but not PI3K/AKT signaling.

Abstract: Importantly, the expression of activated RAF-1 is sufficient to rescue microtubule nucleation inhibited by the mutant integrin by promoting the centrosomal accumulation of γ-tubulin.

Abstract: Our data define a novel paradigm of integrin signaling, where integrins regulate microtubule nucleation by promoting the formation of androgen receptor-Src signaling complexes to activate the MEK/ERK signaling pathway.


Consumption of added sugars from liquid but not solid sources predicts impaired glucose homeostasis and insulin resistance among youth at risk of obesity.

Abstract: Little is known about longitudinal associations between added sugar consumption (solid and liquid sources) and glucose-insulin homeostasis among youth.

Abstract: Caucasian children (8-10 y) with at least one obese biological parent were recruited in the QUébec Adipose and Lifestyle InvesTigation in Youth (QUALITY) cohort (n = 630) and followed-up 2 y later (n = 564).

Abstract: Added sugars were assessed by 3 24-h dietary recalls at baseline.

Abstract: Two-year changes were examined in multivariate linear regression models, adjusting for baseline level, age, sex, Tanner stage, energy intake, fat mass (dual-energy X-ray absorptiometry), and physical activity (7 d accelerometer).

Abstract: Added sugar intake in either liquid or solid sources was not related to changes in adiposity measures (fat mass, body mass index, or waist circumference).

Abstract: However, a higher consumption (10 g/d) of added sugars from liquid sources was associated with 0.04 mmol/L higher fasting glucose, 2.3 pmol/L higher fasting insulin, 0.1 unit higher homeostasis model assessment of insulin resistance (HOMA-IR), and 0.4 unit lower Matsuda-insulin sensitivity index (Matsuda-ISI) in all participants (P < 0.01).

Abstract: No associations were observed with consumption of added sugars from solid sources.

Abstract: Overweight/obese children at baseline had greater increases in adiposity indicators, fasting insulin, and HOMA-IR and decreases in Matsuda-ISI during those 2 y than normal-weight children.

Abstract: Consumption of added sugars from liquid or solid sources was not associated with changes in adiposity, but liquid added sugars were a risk factor for the development of impaired glucose homeostasis and insulin resistance over 2 y among youth at risk of obesity.


Towards an improved investment approach for an effective response to HIV/AIDS.

Abstract: Substantial changes are needed to achieve a more targeted and strategic approach to investment in the response to the HIV/AIDS epidemic that will yield long-term dividends.

Abstract: Until now, advocacy for resources has been done on the basis of a commodity approach that encouraged scaling up of numerous strategies in parallel, irrespective of their relative effects.

Abstract: We propose a strategic investment framework that is intended to support better management of national and international HIV/AIDS responses than exists with the present system.

Abstract: Our framework incorporates major efficiency gains through community mobilisation, synergies between programme elements, and benefits of the extension of antiretroviral therapy for prevention of HIV transmission.

Abstract: It proposes three categories of investment, consisting of six basic programmatic activities, interventions that create an enabling environment to achieve maximum effectiveness, and programmatic efforts in other health and development sectors related to HIV/AIDS.

Abstract: The yearly cost of achievement of universal access to HIV prevention, treatment, care, and support by 2015 is estimated at no less than US$22 billion.

Abstract: Implementation of the new investment framework would avert 12·2 million new HIV infections and 7·4 million deaths from AIDS between 2011 and 2020 compared with continuation of present approaches, and result in 29·4 million life-years gained.

Abstract: The framework is cost effective at $1060 per life-year gained, and the additional investment proposed would be largely offset from savings in treatment costs alone.


Pivotal Roles of T-Helper 17-Related Cytokines, IL-17, IL-22, and IL-23, in Inflammatory Diseases

Abstract: T-helper 17 (Th17) cells are characterized by producing interleukin-17 (IL-17, also called IL-17A), IL-17F, IL-21, and IL-22 and potentially TNF- α and IL-6 upon certain stimulation.

Abstract: IL-23, which promotes Th17 cell development, as well as IL-17 and IL-22 produced by the Th17 cells plays essential roles in various inflammatory diseases, such as experimental autoimmune encephalomyelitis, rheumatoid arthritis, colitis, and Concanavalin A-induced hepatitis.

Abstract: In this review, we summarize the characteristics of the functional role of Th17 cells, with particular focus on the Th17 cell-related cytokines such as IL-17, IL-22, and IL-23, in mouse models and human inflammatory diseases.


LXRS and FXR: the yin and yang of cholesterol and fat metabolism.

Abstract: Liver X receptors (LXRs) and farnesoid X receptor (FXR) are nuclear receptors that function as intracellular sensors for sterols and bile acids, respectively.

Abstract: In response to their ligands, these receptors induce transcriptional responses that maintain a balanced, finely tuned regulation of cholesterol and bile acid metabolism.

Abstract: LXRs also permit the efficient storage of carbohydrate- and fat-derived energy, whereas FXR activation results in an overall decrease in triglyceride levels and modulation of glucose metabolism.

Abstract: The elegant, dual interplay between these two receptor systems suggests that they coevolved to constitute a highly sensitive and efficient system for the maintenance of total body fat and cholesterol homeostasis.

Abstract: Emerging evidence suggests that the tissue-specific action of these receptors is also crucial for the proper function of the cardiovascular, immune, reproductive, endocrine pancreas, renal, and central nervous systems.

Abstract: Together, LXRs and FXR represent potential therapeutic targets for the treatment and prevention of numerous metabolic and lipid-related diseases.


Kaposi's sarcoma-associated herpesvirus/human herpesvirus type 8-positive solid lymphomas: a tissue-based variant of primary effusion lymphoma.

Abstract: Kaposi's sarcoma-associated herpesvirus (KSHV), also termed human herpesvirus type 8, is consistently identified in Kaposi's sarcoma, primary effusion lymphoma (PEL), and multicentric Castleman's disease.

Abstract: Here we report four cases of KSHV-bearing solid lymphomas that occurred in AIDS patients (cases 1 to 3) and in a human immunodeficiency virus (HIV)-seronegative person (case 4).

Abstract: The patients presented extranodal masses in the abdomen (cases 1, 3, and 4) or skin (case 2), and nodal involvement, together with Kaposi's sarcoma (case 3).

Abstract: The gastrointestinal tract was involved in two patients (cases 1 and 3).

Abstract: The patients did not develop a lymphomatous effusion.

Abstract: KSHV was detected in the tumor cells of all cases by immunohistochemistry and by polymerase chain reaction.

Abstract: Epstein-Barr virus was detected in two of the HIV-related cases.

Abstract: All KSHV-positive solid lymphomas exhibited PEL-like cell morphology.

Abstract: To investigate the relationship of these disorders to PEL and to other AIDS-associated diffuse large cell lymphomas, KSHV-positive solid lymphomas were tested for the expression of a set of genes that were previously shown by gene profiling analysis to define PEL tumor cells.

Abstract: The results showed that expression of this set of genes in KSHV-positive lymphomas is similar to that of PEL but distinct from KSHV-negative AIDS-associated diffuse large cell lymphomas.

Abstract: Because pathobiological features of KSHV-positive solid lymphomas closely mimic those of PEL, our results suggest that KSHV-positive solid lymphomas should be considered as a tissue-based variant of classical PEL, irrespective of HIV status.


STAR: ultrafast universal RNA-seq aligner.

Abstract: MOTIVATION Accurate alignment of high-throughput RNA-seq data is a challenging and yet unsolved problem because of the non-contiguous transcript structure, relatively short read lengths and constantly increasing throughput of the sequencing technologies.

Abstract: Currently available RNA-seq aligners suffer from high mapping error rates, low mapping speed, read length limitation and mapping biases.

Abstract: RESULTS To align our large (>80 billon reads) ENCODE Transcriptome RNA-seq dataset, we developed the Spliced Transcripts Alignment to a Reference (STAR) software based on a previously undescribed RNA-seq alignment algorithm that uses sequential maximum mappable seed search in uncompressed suffix arrays followed by seed clustering and stitching procedure.

Abstract: STAR outperforms other aligners by a factor of >50 in mapping speed, aligning to the human genome 550 million 2 × 76 bp paired-end reads per hour on a modest 12-core server, while at the same time improving alignment sensitivity and precision.

Abstract: In addition to unbiased de novo detection of canonical junctions, STAR can discover non-canonical splices and chimeric (fusion) transcripts, and is also capable of mapping full-length RNA sequences.

Abstract: Using Roche 454 sequencing of reverse transcription polymerase chain reaction amplicons, we experimentally validated 1960 novel intergenic splice junctions with an 80-90% success rate, corroborating the high precision of the STAR mapping strategy.

Abstract: AVAILABILITY AND IMPLEMENTATION STAR is implemented as a standalone C++ code.

Abstract: STAR is free open source software distributed under GPLv3 license and can be downloaded from http://code.google.com/p/rna-star/.


Effect of alendronate on risk of fracture in women with low bone density but without vertebral fractures: results from the Fracture Intervention Trial.

Abstract: CONTEXT Alendronate sodium reduces fracture risk in postmenopausal women who have vertebral fractures, but its effects on fracture risk have not been studied for women without vertebral fractures.

Abstract: OBJECTIVE To test the hypothesis that 4 years of alendronate would decrease the risk of clinical and vertebral fractures in women who have low bone mineral density (BMD) but no vertebral fractures.

Abstract: DESIGN Randomized, blinded, placebo-controlled trial.

Abstract: SETTING Eleven community-based clinical research centers.

Abstract: SUBJECTS Women aged 54 to 81 years with a femoral neck BMD of 0.68 g/cm2 or less (Hologic Inc, Waltham, Mass) but no vertebral fracture; 4432 were randomized to alendronate or placebo and 4272 (96%) completed outcome measurements at the final visit (an average of 4.2 years later).

Abstract: INTERVENTION All participants reporting calcium intakes of 1000 mg/d or less received a supplement containing 500 mg of calcium and 250 IU of cholecalciferol.

Abstract: Subjects were randomly assigned to either placebo or 5 mg/d of alendronate sodium for 2 years followed by 10 mg/d for the remainder of the trial.

Abstract: MAIN OUTCOME MEASURES Clinical fractures confirmed by x-ray reports, new vertebral deformities detected by morphometric measurements on radiographs, and BMD measured by dual x-ray absorptiometry.

Abstract: RESULTS Alendronate increased BMD at all sites studied (P<.001) and reduced clinical fractures from 312 in the placebo group to 272 in the intervention group, but not significantly so (14% reduction; relative hazard [RH], 0.86; 95% confidence interval [CI], 0.73-1.01).

Abstract: Alendronate reduced clinical fractures by 36% in women with baseline osteoporosis at the femoral neck (>2.5 SDs below the normal young adult mean; RH, 0.64; 95% CI, 0.50-0.82; treatment-control difference, 6.5%; number needed to treat [NNT], 15), but there was no significant reduction among those with higher BMD (RH, 1.08; 95% CI, 0.87-1.35).

Abstract: Alendronate decreased the risk of radiographic vertebral fractures by 44% overall (relative risk, 0.56; 95% CI, 0.39-0.80; treatment-control difference, 1.7%; NNT, 60).

Abstract: Alendronate did not increase the risk of gastrointestinal or other adverse effects.

Abstract: CONCLUSIONS In women with low BMD but without vertebral fractures, 4 years of alendronate safely increased BMD and decreased the risk of first vertebral deformity.

Abstract: Alendronate significantly reduced the risk of clinical fractures among women with osteoporosis but not among women with higher BMD.


AP-1/sigma1B-adaptin mediates endosomal synaptic vesicle recycling, learning and memory.

Abstract: Synaptic vesicle recycling involves AP-2/clathrin-mediated endocytosis, but it is not known whether the endosomal pathway is also required.

Abstract: Mice deficient in the tissue-specific AP-1-sigma1B complex have impaired synaptic vesicle recycling in hippocampal synapses.

Abstract: The ubiquitously expressed AP-1-sigma1A complex mediates protein sorting between the trans-Golgi network and early endosomes.

Abstract: Vertebrates express three sigma1 subunit isoforms: A, B and C. The expressions of sigma1A and sigma1B are highest in the brain.

Abstract: Synaptic vesicle reformation in cultured neurons from sigma1B-deficient mice is reduced upon stimulation, and large endosomal intermediates accumulate.

Abstract: The sigma1B-deficient mice have reduced motor coordination and severely impaired long-term spatial memory.

Abstract: These data reveal a molecular mechanism for a severe human X-chromosome-linked mental retardation.


Update of the FANTOM web resource: high resolution transcriptome of diverse cell types in mammals

Abstract: Upon the first publication of the fifth iteration of the Functional Annotation of Mammalian Genomes collaborative project, FANTOM5, we gathered a series of primary data and database systems into the FANTOM web resource (http://fantom.gsc.riken.jp) to facilitate researchers to explore transcriptional regulation and cellular states.

Abstract: In the course of the collaboration, primary data and analysis results have been expanded, and functionalities of the database systems enhanced.

Abstract: We believe that our data and web systems are invaluable resources, and we think the scientific community will benefit for this recent update to deepen their understanding of mammalian cellular organization.

Abstract: We introduce the contents of FANTOM5 here, report recent updates in the web resource and provide future perspectives.


Adverse Events Associated With the Treatment of Multidrug-Resistant Tuberculosis: A Systematic Review and Meta-analysis.

Abstract: Multidrug-resistant tuberculosis (MDR-TB) is a growing public health problem.

Abstract: Due to long duration of therapy and concurrent use of multiple second-line drugs, adverse drug events (ADEs) are regarded as the most important clinical consideration in patients undergoing anti-MDR-TB treatment.

Abstract: To evaluate the frequency and type of treatment-related ADEs owing to MDR-TB therapy.

Abstract: The Cochrane Library, MEDLINE, and EMBASE were searched from inception through October 1, 2012, with additional manual search of International Journal of Tuberculosis and Lung Disease.

Abstract: Studies with available ADEs were selected if MDR-TB patients were treated with regimen including second-line drugs.

Abstract: Pooled estimations of incidence for each specific type of ADEs were calculated with 95% confidence intervals using random-effects model.

Abstract: Of the 5346 patients included, 2602 (57.3%) experienced at least 1 kind of ADE.

Abstract: The 3 most common side effects were gastrointestinal disorders (32.1%), ototoxicity (14.6%), and psychiatric disorders (13.2%).

Abstract: Subgroup analyses based on each characteristic (study population, previous tuberculosis treated, human immunodeficiency virus prevalence, and length of treatment) did not show any significant difference between groups.

Abstract: Additionally, among 1519 patients who developed ADEs with available data of impact on MDR-TB therapy, 70.4% required change of MDR-TB treatment.

Abstract: Adverse events were common among MDR-TB cases, occurring in more than half of the cases, with over two-thirds requiring change of anti-MDR-TB treatment.

Abstract: MDR-TB patients should be monitored closely and managed aggressively for side effects during therapy, especially for ototoxicity and psychiatric disorders.


Presynaptically Localized Cyclic GMP-Dependent Protein Kinase 1 Is a Key Determinant of Spinal Synaptic Potentiation and Pain Hypersensitivity

Abstract: Synaptic long-term potentiation (LTP) at spinal neurons directly communicating pain-specific inputs from the periphery to the brain has been proposed to serve as a trigger for pain hypersensitivity in pathological states.

Abstract: Previous studies have functionally implicated the NMDA receptor-NO pathway and the downstream second messenger, cGMP, in these processes.

Abstract: Because cGMP can broadly influence diverse ion-channels, kinases, and phosphodiesterases, pre- as well as post-synaptically, the precise identity of cGMP targets mediating spinal LTP, their mechanisms of action, and their locus in the spinal circuitry are still unclear.

Abstract: Here, we found that Protein Kinase G1 (PKG-I) localized presynaptically in nociceptor terminals plays an essential role in the expression of spinal LTP.

Abstract: Using the Cre-lox P system, we generated nociceptor-specific knockout mice lacking PKG-I specifically in presynaptic terminals of nociceptors in the spinal cord, but not in post-synaptic neurons or elsewhere (SNS-PKG-I(-/-) mice).

Abstract: Patch clamp recordings showed that activity-induced LTP at identified synapses between nociceptors and spinal neurons projecting to the periaqueductal grey (PAG) was completely abolished in SNS-PKG-I(-/-) mice, although basal synaptic transmission was not affected.

Abstract: Analyses of synaptic failure rates and paired-pulse ratios indicated a role for presynaptic PKG-I in regulating the probability of neurotransmitter release.

Abstract: Inositol 1,4,5-triphosphate receptor 1 and myosin light chain kinase were recruited as key phosphorylation targets of presynaptic PKG-I in nociceptive neurons.

Abstract: Finally, behavioural analyses in vivo showed marked defects in SNS-PKG-I(-/-) mice in several models of activity-induced nociceptive hypersensitivity, and pharmacological studies identified a clear contribution of PKG-I expressed in spinal terminals of nociceptors.

Abstract: Our results thus indicate that presynaptic mechanisms involving an increase in release probability from nociceptors are operational in the expression of synaptic LTP on spinal-PAG projection neurons and that PKG-I localized in presynaptic nociceptor terminals plays an essential role in this process to regulate pain sensitivity.


A Sleeping Beauty mutagenesis screen reveals a tumor suppressor role for Ncoa2/Src-2 in liver cancer.

Abstract: The Sleeping Beauty (SB) transposon mutagenesis system is a powerful tool that facilitates the discovery of mutations that accelerate tumorigenesis.

Abstract: In this study, we sought to identify mutations that cooperate with MYC, one of the most commonly dysregulated genes in human malignancy.

Abstract: We performed a forward genetic screen with a mouse model of MYC-induced liver cancer using SB-mediated mutagenesis.

Abstract: We sequenced insertions in 63 liver tumor nodules and identified at least 16 genes/loci that contribute to accelerated tumor development.

Abstract: RNAi-mediated knockdown in a liver progenitor cell line further validate three of these genes, Ncoa2/Src-2, Zfx, and Dtnb, as tumor suppressors in liver cancer.

Abstract: Moreover, deletion of Ncoa2/Src-2 in mice predisposes to diethylnitrosamine-induced liver tumorigenesis.

Abstract: These findings reveal genes and pathways that functionally restrain MYC-mediated liver tumorigenesis and therefore may provide targets for cancer therapy.


Protein-Protein Interaction Inhibition (2P2I)-Oriented Chemical Library Accelerates Hit Discovery.

Abstract: Protein-protein interactions (PPIs) represent an enormous source of opportunity for therapeutic intervention.

Abstract: We and others have recently pinpointed key rules that will help in identifying the next generation of innovative drugs to tackle this challenging class of targets within the next decade.

Abstract: We used these rules to design an oriented chemical library corresponding to a set of diverse "PPI-like" modulators with cores identified as privileged structures in therapeutics.

Abstract: In this work, we purchased the resulting 1664 structurally diverse compounds and evaluated them on a series of representative protein-protein interfaces with distinct "druggability" potential using homogeneous time-resolved fluorescence (HTRF) technology.

Abstract: For certain PPI classes, analysis of the hit rates revealed up to 100 enrichment factors compared with nonoriented chemical libraries.

Abstract: This observation correlates with the predicted "druggability" of the targets.

Abstract: A specific focus on selectivity profiles, the three-dimensional (3D) molecular modes of action resolved by X-ray crystallography, and the biological activities of identified hits targeting the well-defined "druggable" bromodomains of the bromo and extraterminal (BET) family are presented as a proof-of-concept.

Abstract: Overall, our present study illustrates the potency of machine learning-based oriented chemical libraries to accelerate the identification of hits targeting PPIs.

Abstract: A generalization of this method to a larger set of compounds will accelerate the discovery of original and potent probes for this challenging class of targets.


Molecular characterization of endometrial cancer: a correlative study assessing microsatellite instability, MLH1 hypermethylation, DNA mismatch repair protein expression, and PTEN, PIK3CA, KRAS, and BRAF mutation analysis.

Abstract: Endometrial cancer is associated with numeric and structural chromosomal abnormalities, microsatellite instability (MSI), and alterations that activate oncogenes and inactivate tumor suppressor genes.

Abstract: The aim of this study was to characterize a set of endometrial cancers using multiple molecular genetic and immunohistochemical techniques.

Abstract: Ninety-six cases were examined for genomic alterations by MSI, MLH1 promoter hypermethylation, p53 and mismatch repair protein expression (MLH1, MSH2, MSH6, PMS2), and PTEN, PIK3CA, KRAS, and BRAF mutation analysis.

Abstract: At least 1 alteration was identified in 48 of 87 (55%) specimens tested for PTEN, making it the most common abnormality in this study.

Abstract: A PIK3CA alteration was observed in 16 (17%) specimens.

Abstract: Twenty-nine of 94 (31%) MSI tested tumors exhibited an MSI-H phenotype.

Abstract: Of the 29 MSI-H cases, 24 (83%) were positive for methylation of the MLH1 promoter region.

Abstract: Twenty-three (82%) of the 28 MSI-H cases with immunohistochemistry results showed loss of expression of MLH1/PMS2 (n=19), MSH2/MSH6 (n=2), or MSH6 only (n=2).

Abstract: Of the 19 MSI-H cases with loss of MLH1/PMS2 on immunohistochemistry, 18 were positive, and 1 was equivocal for MLH1 promoter hypermethylation.

Abstract: Twelve of 94 cases (13%) analyzed for KRAS mutations were found to have a mutation.

Abstract: No BRAF V600E mutations were indentified.

Abstract: This study provides a comprehensive molecular genetic analysis of commonly analyzed targets in a large cohort of endometrial cancers.


Identification of indole-3-carboxylic acid as mediator of priming against Plectosphaerella cucumerina.

Abstract: Plant resistance against the necrotrophic pathogen Plectosphaerella cucumerina is mediated by a combination of several hormonal-controlled signalling pathways.

Abstract: The priming agent β-aminobutyric acid (BABA) is able to induce effective resistance against this pathogen by stimulating callose-rich cell wall depositions.

Abstract: In the present research it is demonstrated that BABA-Induced Resistance (BABA-IR) against P. cucumerina in Arabidopsis has additional components such as the induction of defences mediated by indolic derivatives.

Abstract: Chromatographic approach for the detection and characterization of metabolites enhanced by BABA compared with water-treated plants only when the challenge is present has been developed.

Abstract: The metabolites matching this criteria are considered to be primed by BABA.

Abstract: The analytic procedure is based on the combination of liquid chromatography (LC) with a triple quadrupole (TQD) detector in a precursor ion scanning mode.

Abstract: Using this analytical system a signal in negative electro-spray mode of 160 m/z is primed by BABA in infected plants.

Abstract: A subsequent exact mass analysis in a quadrupole time-of-flight mass spectrometer demonstrated that this ion was the indole-derivative metabolite indole-3-carboxylic acid (I3CA).

Abstract: The identity of indole-3-carboxilic acid was definitively confirmed by comparing its retention time and fragmentation spectra with a commercial standard.

Abstract: Quantification of I3CA in primed plants showed that this indolic metabolite is specifically primed by BABA upon P. cucumerina infection, while other indolic compounds such as IAA and camalexin are not.

Abstract: Taking together these observations with the known role of callose in priming against this pathogen, suggests that priming is not a single mechanism but rather a multicomponent defence.


Evaluation of networks of randomized trials.

Abstract: Randomized trials may be designed and interpreted as single experiments or they may be seen in the context of other similar or relevant evidence.

Abstract: The amount and complexity of available randomized evidence vary for different topics.

Abstract: Systematic reviews may be useful in identifying gaps in the existing randomized evidence, pointing to discrepancies between trials, and planning future trials.

Abstract: A new, promising, but also very much debated extension of systematic reviews, mixed treatment comparison (MTC) meta-analysis, has become increasingly popular recently.

Abstract: MTC meta-analysis may have value in interpreting the available randomized evidence from networks of trials and can rank many different treatments, going beyond focusing on simple pairwise-comparisons.

Abstract: Nevertheless, the evaluation of networks also presents special challenges and caveats.

Abstract: In this article, we review the statistical methodology for MTC meta-analysis.

Abstract: We discuss the concept of inconsistency and methods that have been proposed to evaluate it as well as the methodological gaps that remain.

Abstract: We introduce the concepts of network geometry and asymmetry, and propose metrics for the evaluation of the asymmetry.

Abstract: Finally, we discuss the implications of inconsistency, network geometry and asymmetry in informing the planning of future trials.


Metabolic regulation by the mitochondrial phosphatase PTPMT1 is required for hematopoietic stem cell differentiation.

Abstract: The regulation and coordination of mitochondrial metabolism with hematopoietic stem cell (HSC) self-renewal and differentiation is not fully understood.

Abstract: Here we report that depletion of PTPMT1, a PTEN-like mitochondrial phosphatase, in inducible or hematopoietic-cell-specific knockout mice resulted in hematopoietic failure due to changes in the cell cycle and a block in the differentiation of HSCs.

Abstract: Surprisingly, the HSC pool was increased by ∼40-fold in PTPMT1 knockout mice.

Abstract: Reintroduction of wild-type PTPMT1, but not catalytically deficient PTPMT1 or truncated PTPMT1 lacking mitochondrial localization, restored differentiation capabilities of PTPMT1 knockout HSCs.

Abstract: Further analyses demonstrated that PTPMT1 deficiency altered mitochondrial metabolism and that phosphatidylinositol phosphate substrates of PTPMT1 directly enhanced fatty-acid-induced activation of mitochondrial uncoupling protein 2.

Abstract: Intriguingly, depletion of PTPMT1 from myeloid, T lymphoid, or B lymphoid progenitors did not cause any defects in lineage-specific knockout mice.

Abstract: This study establishes a crucial role of PTPMT1 in the metabolic regulation of HSC function.


GSK-3-mediated phosphorylation enhances Maf-transforming activity.

Abstract: The Maf oncoproteins are b-Zip transcription factors of the AP-1 superfamily.

Abstract: They are involved in developmental, metabolic, and tumorigenic processes.

Abstract: Maf proteins are overexpressed in about 50% of human multiple myelomas.

Abstract: Here, we show that Maf-transforming activity is controlled by GSK-3-dependent phosphorylation and that phosphorylation by GSK-3 can increase the oncogenic activity of a protein.

Abstract: Using microarray analysis, we identify a gene-expression subprogram regulated by GSK-3-mediated Maf phosphorylation involved in extracellular matrix remodeling and relevant to cancer progression.

Abstract: We also demonstrate that GSK-3 triggers MafA sequential phosphorylation on residues S61, T57, T53, and S49, inducing its ubiquitination and degradation.

Abstract: Paradoxically, this phosphorylation increases MafA-transcriptional activity through the recruitment of the coactivator P/CAF.

Abstract: We further demonstrate that P/CAF protects MafA from ubiquitination and degradation, suggesting that, upon the release of the coactivator complex, MafA becomes polyubiquitinated and degraded to allow the response to terminate.


NETs are a source of citrullinated autoantigens and stimulate inflammatory responses in rheumatoid arthritis.

Abstract: The early events leading to the development of rheumatoid arthritis (RA) remain unclear, but formation of autoantibodies to citrullinated protein antigens (ACPAs) is considered a key pathogenic event.

Abstract: Neutrophils isolated from patients with various autoimmune diseases display enhanced neutrophil extracellular trap (NET) formation, a phenomenon that exposes autoantigens in the context of immunostimulatory molecules.

Abstract: We investigated whether aberrant NETosis occurs in RA, determined its triggers, and examined its deleterious inflammatory consequences.

Abstract: Enhanced NETosis was observed in circulating and RA synovial fluid neutrophils compared to neutrophils from healthy controls and from patients with osteoarthritis (OA).

Abstract: Further, netting neutrophils infiltrated RA synovial tissue, rheumatoid nodules, and skin.

Abstract: NETosis correlated with ACPA presence and levels and with systemic inflammatory markers.

Abstract: RA sera and immunoglobulin fractions from RA patients with high levels of ACPA and/or rheumatoid factor significantly enhanced NETosis, and the NETs induced by these autoantibodies displayed distinct protein content.

Abstract: Indeed, during NETosis, neutrophils externalized the citrullinated autoantigens implicated in RA pathogenesis, and anti-citrullinated vimentin antibodies potently induced NET formation.

Abstract: Moreover, the inflammatory cytokines interleukin-17A (IL-17A) and tumor necrosis factor-α (TNF-α) induced NETosis in RA neutrophils.

Abstract: In turn, NETs significantly augmented inflammatory responses in RA and OA synovial fibroblasts, including induction of IL-6, IL-8, chemokines, and adhesion molecules.

Abstract: These observations implicate accelerated NETosis in RA pathogenesis, through externalization of citrullinated autoantigens and immunostimulatory molecules that may promote aberrant adaptive and innate immune responses in the joint and in the periphery, and perpetuate pathogenic mechanisms in this disease.


Is autoimmunity the Achilles' heel of cancer immunotherapy?

Abstract: The emergence of immuno-oncology as the first broadly successful strategy for metastatic cancer will require clinicians to integrate this new pillar of medicine with chemotherapy, radiation, and targeted small-molecule compounds.

Abstract: Of equal importance is gaining an understanding of the limitations and toxicities of immunotherapy.

Abstract: Immunotherapy was initially perceived to be a relatively less toxic approach to cancer treatment than other available therapies—and surely it is, when compared to those.

Abstract: However, as the use of immunotherapy becomes more common, especially as first- and second-line treatments, immunotoxicity and autoimmunity are emerging as the Achilles' heel of immunotherapy.

Abstract: In this Perspective, we discuss evidence that the occurrence of immunotoxicity bodes well for the patient, and describe mechanisms that might be related to the induction of autoimmunity.

Abstract: We then explore approaches to limit immunotoxicity, and discuss the future directions of research and reporting that are needed to diminish it.


Trophoblast differentiation defect in human embryonic stem cells lacking PIG-A and GPI-anchored cell-surface proteins.

Abstract: Pluripotent human embryonic stem (hES) cells can differentiate into various cell types derived from the three embryonic germ layers and extraembryonic tissues such as trophoblasts.

Abstract: The mechanisms governing lineage choices of hES cells are largely unknown.

Abstract: Here, we report that we established two independent hES cell clones lacking a group of cell surface molecules, glycosyl-phosphatidyl-inositol-anchored proteins (GPI-APs).

Abstract: The GPI-AP deficiency in these two hES clones is due to the deficiency in the gene expression of PIG-A (phosphatidyl-inositol-glycan class A), which is required for the first step of GPI synthesis.

Abstract: GPI-AP-deficient hES cells were capable of forming embryoid bodies and initiating cell differentiation into the three embryonic germ layers.

Abstract: However, GPI-AP-deficient hES cells failed to form trophoblasts after differentiation induction by embryoid body formation or by adding exogenous BMP4.

Abstract: The defect in trophoblast formation was due to the lack of GPI-anchored BMP coreceptors, resulting in the impairment of full BMP4 signaling activation in the GPI-AP-deficient hES cells.

Abstract: These data reveal that GPI-AP-enhanced full activation of BMP signaling is required for human trophoblast formation.


CRISPR Interference Limits Horizontal Gene Transfer in Staphylococci by Targeting DNA

Abstract: Horizontal gene transfer (HGT) in bacteria and archaea occurs through phage transduction, transformation, or conjugation, and the latter is particularly important for the spread of antibiotic resistance.

Abstract: Clustered, regularly interspaced, short palindromic repeat (CRISPR) loci confer sequence-directed immunity against phages.

Abstract: A clinical isolate of Staphylococcus epidermidis harbors a CRISPR spacer that matches the nickase gene present in nearly all staphylococcal conjugative plasmids.

Abstract: Here we show that CRISPR interference prevents conjugation and plasmid transformation in S. epidermidis.

Abstract: Insertion of a self-splicing intron into nickase blocks interference despite the reconstitution of the target sequence in the spliced mRNA, which indicates that the interference machinery targets DNA directly.

Abstract: We conclude that CRISPR loci counteract multiple routes of HGT and can limit the spread of antibiotic resistance in pathogenic bacteria.


Development of leukemia in donor cells after allogeneic stem cell transplantation--a survey of the European Group for Blood and Marrow Transplantation (EBMT).

Abstract: Leukemia in donor cells (donor cell leukemia; DCL) has been reported as a rare but severe complication of allogeneic stem cell transplantation (SCT).

Abstract: However, the incidence, potential pathogenetic factors, therapeutic options and outcome of patients suffering from DCL and the leukemia risk of their donors are not well defined.

Abstract: A questionnaire survey was carried out within European Blood and Marrow Transplantation Group (EBMT) centers.

Abstract: Ninety-one EBMT centers participated in this survey, covering 10489 allogeneic SCT between 12/1982 and 09/2003.

Abstract: Fourteen cases of DCL, most with a myeloid phenotype (7 cases of acute myeloid leukemia, 3 each of acute lymphocytic leukemia and 1 case of chronic myeloid leukemia) were identified.

Abstract: Demonstration of donor cell origin included molecular analysis of chimerism in most cases.

Abstract: DCL type and cytogenetic alterations were independent from the original disease.

Abstract: The median time between transplantation and diagnosis of DCL was 17 months (4-164).

Abstract: No type of conditioning, donor, graft manipulation, graft-versus-host disease prophylaxis or subsequent complications were identified as risk factors for DCL.

Abstract: Chemotherapy induced remissions in DCL and 2 of 5 patients remain alive in remission after a second transplant.

Abstract: None of the stem cell donors developed hematologic malignancies (median follow-up period of 9 years; range 6-30 years).

Abstract: DCL is an extremely rare complication of allogeneic SCT in which treatment attempts with chemotherapy and a second SCT are justified.

Abstract: Donors are not at an increased risk of developing hematologic malignancies.


Lack of evidence of transfusion transmission of Creutzfeldt-Jakob disease in a US surveillance study.

Abstract: BACKGROUND Since 2004, several reported transfusion transmissions of variant Creutzfeldt-Jakob disease (vCJD) in the United Kingdom have reawakened concerns about the possible risk of similar transmissions of nonvariant or classic forms of CJD.

Abstract: STUDY DESIGN AND METHODS Patients with a CJD diagnosis and a history of donating blood were reported to the study coordinator.

Abstract: Through review of blood distribution and hospital records, the recipients of blood components from these donors were identified.

Abstract: We then determined each recipient's vital status and, if deceased, the cause(s) of death identified by matching the recipient's personal identifiers with the Centers for Disease Control and Prevention's National Death Index database.

Abstract: We conducted such searches after recipients were enrolled in this study and annually thereafter for those who remained alive.

Abstract: RESULTS The study included a total of 36 blood donors who subsequently developed CJD and 436 recipients.

Abstract: Through 2006, 91 of these recipients were still alive, 329 were deceased, and 16 were lost to follow-up.

Abstract: After transfusion, these three groups had survived a total of 2096.0 person-years.

Abstract: A total of 144 recipients survived 5 years or longer after transfusion and 68 of them had received blood donated 60 or fewer months before the onset of CJD in the donor.

Abstract: We identified no recipient with CJD.

Abstract: CONCLUSIONS The current results of this large, ongoing lookback study show no evidence of transfusion transmission of CJD.

Abstract: They reinforce the conclusion that the risk, if any, of transfusion transmission of prion disease by CJD donors is significantly lower than the comparable risk of such transmission by vCJD donors.


Large-scale hypomethylated blocks associated with Epstein-Barr virus-induced B-cell immortalization.

Abstract: Altered DNA methylation occurs ubiquitously in human cancer from the earliest measurable stages.

Abstract: A cogent approach to understanding the mechanism and timing of altered DNA methylation is to analyze it in the context of carcinogenesis by a defined agent.

Abstract: Epstein-Barr virus (EBV) is a human oncogenic herpesvirus associated with lymphoma and nasopharyngeal carcinoma, but also used commonly in the laboratory to immortalize human B-cells in culture.

Abstract: Here we have performed whole-genome bisulfite sequencing of normal B-cells, activated B-cells, and EBV-immortalized B-cells from the same three individuals, in order to identify the impact of transformation on the methylome.

Abstract: Surprisingly, large-scale hypomethylated blocks comprising two-thirds of the genome were induced by EBV immortalization but not by B-cell activation per se.

Abstract: These regions largely corresponded to hypomethylated blocks that we have observed in human cancer, and they were associated with gene-expression hypervariability, similar to human cancer, and consistent with a model of epigenomic change promoting tumor cell heterogeneity.

Abstract: We also describe small-scale changes in DNA methylation near CpG islands.

Abstract: These results suggest that methylation disruption is an early and critical step in malignant transformation.


Stem cell self-renewal specified by JAK-STAT activation in response to a support cell cue.

Abstract: Stem cells generate many differentiated, short-lived cell types, such as blood, skin, and sperm, throughout adult life.

Abstract: Stem cells maintain a long-term capacity to divide, producing daughter cells that either self-renew or initiate differentiation.

Abstract: Although the surrounding microenvironment or "niche" influences stem cell fate decisions, few signals that emanate from the niche to specify stem cell self-renewal have been identified.

Abstract: Here we demonstrate that the apical hub cells in the Drosophila testis act as a cellular niche that supports stem cell self-renewal.

Abstract: Hub cells express the ligand Unpaired (Upd), which activates the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway in adjacent germ cells to specify self-renewal and continual maintenance of the germ line stem cell population.


Effects of interventions in pregnancy on maternal weight and obstetric outcomes: meta-analysis of randomised evidence

Abstract: OBJECTIVE To evaluate the effects of dietary and lifestyle interventions in pregnancy on maternal and fetal weight and to quantify the effects of these interventions on obstetric outcomes.

Abstract: DESIGN Systematic review and meta-analysis.

Abstract: DATA SOURCES Major databases from inception to January 2012 without language restrictions.

Abstract: STUDY SELECTION Randomised controlled trials that evaluated any dietary or lifestyle interventions with potential to influence maternal weight during pregnancy and outcomes of pregnancy.

Abstract: DATA SYNTHESIS Results summarised as relative risks for dichotomous data and mean differences for continuous data.

Abstract: RESULTS We identified 44 relevant randomised controlled trials (7278 women) evaluating three categories of interventions: diet, physical activity, and a mixed approach.

Abstract: Overall, there was 1.42 kg reduction (95% confidence interval 0.95 to 1.89 kg) in gestational weight gain with any intervention compared with control.

Abstract: With all interventions combined, there were no significant differences in birth weight (mean difference -50 g, -100 to 0 g) and the incidence of large for gestational age (relative risk 0.85, 0.66 to 1.09) or small for gestational age (1.00, 0.78 to 1.28) babies between the groups, though by itself physical activity was associated with reduced birth weight (mean difference -60 g, -120 to -10 g).

Abstract: Interventions were associated with a reduced the risk of pre-eclampsia (0.74, 0.60 to 0.92) and shoulder dystocia (0.39, 0.22 to 0.70), with no significant effect on other critically important outcomes.

Abstract: Dietary intervention resulted in the largest reduction in maternal gestational weight gain (3.84 kg, 2.45 to 5.22 kg), with improved pregnancy outcomes compared with other interventions.

Abstract: The overall evidence rating was low to very low for important outcomes such as pre-eclampsia, gestational diabetes, gestational hypertension, and preterm delivery.

Abstract: CONCLUSIONS Dietary and lifestyle interventions in pregnancy can reduce maternal gestational weight gain and improve outcomes for both mother and baby.

Abstract: Among the interventions, those based on diet are the most effective and are associated with reductions in maternal gestational weight gain and improved obstetric outcomes.


Tetraspanin 3 Is Required for the Development and Propagation of Acute Myelogenous Leukemia.

Abstract: Acute Myelogenous Leukemia (AML) is an aggressive cancer that strikes both adults and children and is frequently resistant to therapy.

Abstract: Thus, identifying signals needed for AML propagation is a critical step toward developing new approaches for treating this disease.

Abstract: Here, we show that Tetraspanin 3 is a target of the RNA binding protein Musashi 2, which plays a key role in AML.

Abstract: We generated Tspan3 knockout mice that were born without overt defects.

Abstract: However, Tspan3 deletion impaired leukemia stem cell self-renewal and disease propagation and markedly improved survival in mouse models of AML.

Abstract: Additionally, Tspan3 inhibition blocked growth of AML patient samples, suggesting that Tspan3 is also important in human disease.

Abstract: As part of the mechanism, we show that Tspan3 deficiency disabled responses to CXCL12/SDF-1 and led to defects in AML localization within the niche.

Abstract: These identify Tspan3 as an important regulator of aggressive leukemias and highlight a role for Tspan3 in oncogenesis.


Reorganization of enhancer patterns in transition from naive to primed pluripotency.

Abstract: Naive and primed pluripotency is characterized by distinct signaling requirements, transcriptomes, and developmental properties, but both cellular states share key transcriptional regulators: Oct4, Sox2, and Nanog.

Abstract: Here, we demonstrate that transition between these two pluripotent states is associated with widespread Oct4 relocalization, mirrored by global rearrangement of enhancer chromatin landscapes.

Abstract: Our genomic and biochemical analyses identified candidate mediators of primed state-specific Oct4 binding, including Otx2 and Zic2/3.

Abstract: Even when differentiation cues are blocked, premature Otx2 overexpression is sufficient to exit the naive state, induce transcription of a substantial subset of primed pluripotency-associated genes, and redirect Oct4 to previously inaccessible enhancer sites.

Abstract: However, the ability of Otx2 to engage new enhancer regions is determined by its levels, cis-encoded properties of the sites, and the signaling environment.

Abstract: Our results illuminate regulatory mechanisms underlying pluripotency and suggest that the capacity of transcription factors such as Otx2 and Oct4 to pioneer new enhancer sites is highly context dependent.


Topoisomerase II plays an essential role as a swivelase in the late stage of SV40 chromosome replication in vitro.

Abstract: The effects of topoisomerases I and II on the replication of SV40 DNA were examined using an in vitro replication system of purified proteins that constitutes the monopolymerase system.

Abstract: In the presence of the two topoisomerases, two distinct nascent DNAs were formed.

Abstract: One product arising from the replication of the leading template strand was approximately half the size of the template DNA, whereas the other product derived from the lagging template strand consisted of short DNAs.

Abstract: These products were synthesized from both SV40 naked DNA and SV40 chromosomes.

Abstract: For the replication of SV40 naked DNA, either topoisomerase I or II maintained replication fork movement and supported complete leading strand synthesis.

Abstract: When SV40 chromosomes were replicated with the same proteins, reactions containing only topoisomerase I produced shorter leading strands.

Abstract: However, mature size DNA products accumulated in reactions supplemented with topoisomerase II, as well as in reactions containing only topoisomerase II.

Abstract: In the presence of crude extracts of HeLa cells, VP-16, a specific inhibitor of topoisomerase II, blocked elongation of the nascent DNA during the replication of SV40 chromosomes.

Abstract: These results indicate that topoisomerase II plays a crucial role as a swivelase in the late stage of SV40 chromosome replication in vitro.


Bromodomain Proteins Contribute to Maintenance of Bloodstream Form Stage Identity in the African Trypanosome

Abstract: Trypanosoma brucei, the causative agent of African sleeping sickness, is transmitted to its mammalian host by the tsetse.

Abstract: In the fly, the parasite's surface is covered with invariant procyclin, while in the mammal it resides extracellularly in its bloodstream form (BF) and is densely covered with highly immunogenic Variant Surface Glycoprotein (VSG).

Abstract: In the BF, the parasite varies this highly immunogenic surface VSG using a repertoire of ~2500 distinct VSG genes.

Abstract: Recent reports in mammalian systems point to a role for histone acetyl-lysine recognizing bromodomain proteins in the maintenance of stem cell fate, leading us to hypothesize that bromodomain proteins may maintain the BF cell fate in trypanosomes.

Abstract: Using small-molecule inhibitors and genetic mutants for individual bromodomain proteins, we performed RNA-seq experiments that revealed changes in the transcriptome similar to those seen in cells differentiating from the BF to the insect stage.

Abstract: This was recapitulated at the protein level by the appearance of insect-stage proteins on the cell surface.

Abstract: Furthermore, bromodomain inhibition disrupts two major BF-specific immune evasion mechanisms that trypanosomes harness to evade mammalian host antibody responses.

Abstract: First, monoallelic expression of the antigenically varied VSG is disrupted.

Abstract: Second, rapid internalization of antibodies bound to VSG on the surface of the trypanosome is blocked.

Abstract: Thus, our studies reveal a role for trypanosome bromodomain proteins in maintaining bloodstream stage identity and immune evasion.

Abstract: Importantly, bromodomain inhibition leads to a decrease in virulence in a mouse model of infection, establishing these proteins as potential therapeutic drug targets for trypanosomiasis.

Abstract: Our 1.25Å resolution crystal structure of a trypanosome bromodomain in complex with I-BET151 reveals a novel binding mode of the inhibitor, which serves as a promising starting point for rational drug design.


Timing requirements for insulin/IGF-1 signaling in C. elegans.

Abstract: The insulin/IGF-1 (where IGF-1 is insulin-like growth factor-1) signaling pathway influences longevity, reproduction, and diapause in many organisms.

Abstract: Because of the fundamental importance of this system in animal physiology, we asked when during the animal's life it is required to regulate these different processes.

Abstract: We find that in Caenorhabditis elegans, the pathway acts during adulthood, to relatively advanced ages, to influence aging.

Abstract: In contrast, it regulates diapause during development.

Abstract: In addition, the pathway controls longevity and reproduction independently of one another.

Abstract: Together our findings show that life-span regulation can be dissociated temporally from phenotypes that might seem to decrease the quality of life.


Predominance of Type 1 Innate Lymphoid Cells in the Rectal Mucosa of Patients With Non-Celiac Wheat Sensitivity: Reversal After a Wheat-Free Diet

Abstract: OBJECTIVES Non-celiac wheat sensitivity (NCWS) is defined as a reaction to ingested wheat after exclusion of celiac disease and wheat allergy.

Abstract: As its pathogenesis is incompletely understood, we evaluated the inflammatory response in the rectal mucosa of patients with well-defined NCWS.

Abstract: METHODS The prospective study included 22 patients with irritable bowel syndrome (IBS)-like clinical presentation, diagnosed with NCWS by double-blind placebo-controlled challenge.

Abstract: Eight IBS patients not improving on wheat-free diet were used as controls.

Abstract: Two weeks after oral challenge was performed with 80 grams of wheat daily, cells were isolated from rectal biopsies and thoroughly characterized by fluorescence-activated cell sorting analysis for intracellular cytokines and surface markers.

Abstract: RESULTS Rectal biopsies from wheat-challenged NCWS patients showed that a significant mucosal CD45(+) infiltrate consisted of CD3(+) and CD3(-) lymphocytes, with the latter spontaneously producing more interferon (IFN)-γ than IBS controls.

Abstract: About 30% of IFN-γ-producing CD45(+) cells were T-bet(+), CD56(-), NKP44(-), and CD117(-), defining them as a type-1 innate lymphoid cells (ILC1).

Abstract: IFN-γ-producing ILC1 cells significantly decreased in 10 patients analyzed 2 weeks after they resumed a wheat-free diet.

Abstract: CONCLUSIONS These data indicate that, in patients with active NCWS, IFN-γ-producing ILC1 cells infiltrate rectal mucosa and support a role for this innate lymphoid cell population in the pathogenesis of NCWS.


Folding and Self-Assembly of the TatA Translocation Pore Based on a Charge Zipper Mechanism

Abstract: We propose a concept for the folding and self-assembly of the pore-forming TatA complex from the Twin-arginine translocase and of other membrane proteins based on electrostatic "charge zippers.

Abstract: Each subunit of TatA consists of a transmembrane segment, an amphiphilic helix (APH), and a C-terminal densely charged region (DCR).

Abstract: The sequence of charges in the DCR is complementary to the charge pattern on the APH, suggesting that the protein can be "zipped up" by a ladder of seven salt bridges.

Abstract: The length of the resulting hairpin matches the lipid bilayer thickness, hence a transmembrane pore could self-assemble via intra- and intermolecular salt bridges.

Abstract: The steric feasibility was rationalized by molecular dynamics simulations, and experimental evidence was obtained by monitoring the monomer-oligomer equilibrium of specific charge mutants.

Abstract: Similar "charge zippers" are proposed for other membrane-associated proteins, e.g., the biofilm-inducing peptide TisB, the human antimicrobial peptide dermcidin, and the pestiviral E(RNS) protein.


Molecular mechanisms of ribosomal protein gene coregulation.

Abstract: The 137 ribosomal protein genes (RPGs) of Saccharomyces provide a model for gene coregulation.

Abstract: We examined the positional and functional organization of their regulators (Rap1 [repressor activator protein 1], Fhl1, Ifh1, Sfp1, and Hmo1), the transcription machinery (TFIIB, TFIID, and RNA polymerase II), and chromatin at near-base-pair resolution using ChIP-exo, as RPGs are coordinately reprogrammed.

Abstract: Where Hmo1 is enriched, Fhl1, Ifh1, Sfp1, and Hmo1 cross-linked broadly to promoter DNA in an RPG-specific manner and demarcated by general minor groove widening.

Abstract: Importantly, Hmo1 extended 20-50 base pairs (bp) downstream from Fhl1.

Abstract: Upon RPG repression, Fhl1 remained in place.

Abstract: Hmo1 dissociated, which was coupled to an upstream shift of the +1 nucleosome, as reflected by the Hmo1 extension and core promoter region.

Abstract: Fhl1 and Hmo1 may create two regulatable and positionally distinct barriers, against which chromatin remodelers position the +1 nucleosome into either an activating or a repressive state.

Abstract: Consistent with in vitro studies, we found that specific TFIID subunits, in addition to cross-linking at the core promoter, made precise cross-links at Rap1 sites, which we interpret to reflect native Rap1-TFIID interactions.

Abstract: Our findings suggest how sequence-specific DNA binding regulates nucleosome positioning and transcription complex assembly >300 bp away and how coregulation coevolved with coding sequences.


Lipoatrophy and severe metabolic disturbance in mice with fat-specific deletion of PPARγ.

Abstract: Adipose tissue is an important metabolic organ, the dysfunction of which is associated with the development of obesity, diabetes mellitus, and cardiovascular disease.

Abstract: The nuclear receptor peroxisome proliferator-activated receptor gamma (PPARγ) is considered the master regulator of adipocyte differentiation and function.

Abstract: Although its cell-autonomous role in adipogenesis has been clearly demonstrated in cell culture, previous fat-specific knockouts of the murine PPARγ gene did not demonstrate a dramatic phenotype in vivo.

Abstract: Here, using Adipoq-Cre mice to drive adipose-specific recombination, we report a unique fat-specific PPARγ knockout (PPARγ FKO) mouse model with almost no visible brown and white adipose tissue at age 3 mo.

Abstract: As a consequence, PPARγ FKO mice had hugely enlarged pancreatic islets, massive fatty livers, and dramatically elevated levels of blood glucose and serum insulin accompanied by extreme insulin resistance.

Abstract: PPARγ FKO mice also exhibited delayed hair coat formation associated with absence of dermal fat, disrupted mammary gland development with loss of mammary fat pads, and high bone mass with loss of bone marrow fat, indicating the critical roles of adipose PPARγ in these tissues.

Abstract: Together, our data reveal the necessity of fat PPARγ in adipose formation, whole-body metabolic homeostasis, and normal development of fat-containing tissues.


Fluconazole prophylaxis: can we eliminate invasive Candida infections in the neonatal ICU?

Abstract: PURPOSE OF REVIEW Owing to the high mortality, risk of neurodevelopmental impairment and end-organ involvement with fungal infections in the neonate, prevention of invasive Candida infections in extremely preterm infants should be a priority for each neonatal ICU.

Abstract: RECENT FINDINGS Even with prompt or empiric treatment, mortality and neurodevelopmental impairment is high (57%) in infants <1000 g. Multiple studies have been performed with fluconazole prophylaxis, including a recent multicenter randomized controlled trial.

Abstract: All of the studies have demonstrated efficacy and safety with no increase or emergence of fungal resistance.

Abstract: Analysis of these studies demonstrates that fluconazole prophylaxis decreased the incidence of invasive Candida infections in high-risk infants <1000 g by 91% (P = 0.0004) and all infants <1500 g by 85% (P < 0.0001).

Abstract: The mortality rate from all causes was 25% lower (P = 0.029).

Abstract: Furthermore, studies have demonstrated that all Candida-related mortality can be eliminated in an entire neonatal ICU by targeting fluconazole prophylaxis in infants <1000 g.   SUMMARY Targeting fluconazole prophylaxis to infants who are either <1000 g or < or =27 weeks is highly effective, safe and inexpensive, and can eliminate these infections as a cause of neurodevelopmental impairment and mortality.


Fumarase deficiency in dichorionic diamniotic twins.

Abstract: Fumarase deficiency is a rare autosomal recessive inborn error of metabolism of the Krebs Tricarboxylic Acid cycle.

Abstract: A heavy neurological disease burden is imparted by fumarase deficiency, commonly manifesting as microcephaly, dystonia, global developmental delay, seizures, and lethality in the infantile period.

Abstract: Heterozygous carriers also carry an increased risk of developing hereditary leiomyomatosis and renal cell carcinoma.

Abstract: We describe a non-consanguineous family in whom a dichorionic diamniotic twin pregnancy resulted in twin boys with fumarase deficiency proven at the biochemical, enzymatic, and molecular levels.

Abstract: Their clinical phenotype included hepatic involvement.

Abstract: A novel mutation in the fumarate hydratase gene was identified in this family.


Angiopoietin-2 is increased in sepsis and inversely associated with nitric oxide-dependent microvascular reactivity

Abstract: INTRODUCTION Angiopoietin-2 (ang-2), an angiogenic peptide released by endothelial cell Weibel-Palade bodies (WPBs), increases endothelial activation and vascular permeability.

Abstract: Ang-2 is raised in severe sepsis but the mechanisms underlying this are not known.

Abstract: Nitric oxide (NO) inhibits WPB exocytosis, and bioavailability of endothelial NO is decreased in sepsis.

Abstract: We hypothesized that endothelial NO bioavailability would be inversely correlated with ang-2 concentrations in sepsis.

Abstract: METHODS Plasma ang-2, vascular endothelial growth factor (VEGF) and endothelial-active cytokines were assessed in 83 patients with early sepsis and 41 hospital controls, and related to reactive hyperaemia-peripheral arterial tonometry, RH-PAT, a measure of endothelial NO bioavailability.

Abstract: RESULTS Plasma Ang-2 was elevated in sepsis (median [interquartile range (IQR)], ng/ml: severe sepsis 12.4 [8.5-33.4], sepsis without organ failure 6.1 [5.0-10.4], controls 2.7 [2.2-3.6], P < 0.0001).

Abstract: It correlated inversely with RH-PAT (r = -0.38, P < 0.0001) and positively with IL-6 (r = 0.57, P < 0.0001) and degree of organ failure (sequential organ function assessment score) (r = 0.58, P < 0.0001).

Abstract: The correlation of ang-2 with RH-PAT persisted after controlling for sepsis severity.

Abstract: In a longitudinal mixed-effects model, recovery of RH-PAT over time was associated with decline in ang-2.

Abstract: CONCLUSIONS Ang-2 is elevated in proportion to sepsis severity, and inversely correlated with NO-dependent microvascular reactivity.

Abstract: Impaired endothelial NO bioavailability may contribute to increased endothelial cell release of ang-2, endothelial activation and capillary leak.

Abstract: Agents that increase endothelial NO bioavailability or inhibit WPB exocytosis and/or Ang-2 activity may have therapeutic potential in sepsis.


Ecologic versus individual-level sources of bias in ecologic estimates of contextual health effects.

Abstract: A number of authors have attempted to defend ecologic (aggregate) studies by claiming that the goal of those studies is estimation of ecologic (contextual or group-level) effects rather than individual-level effects.

Abstract: Critics of these attempts point out that ecologic effect estimates are inevitably used as estimates of individual effects, despite disclaimers.

Abstract: A more subtle problem is that ecologic variation in the distribution of individual effects can bias ecologic estimates of contextual effects.

Abstract: The conditions leading to this bias are plausible and perhaps even common in studies of ecosocial factors and health outcomes because social context is not randomized across typical analysis units (administrative regions).

Abstract: By definition, ecologic data contain only marginal observations on the joint distribution of individually defined confounders and outcomes, and so identify neither contextual nor individual-level effects.

Abstract: While ecologic studies can still be useful given appropriate caveats, their problems are better addressed by multilevel study designs, which obtain and use individual as well as group-level data.

Abstract: Nonetheless, such studies often share certain special problems with ecologic studies, including problems due to inappropriate aggregation and problems due to temporal changes in covariate distributions.


Left–right asymmetry in the vertebrate embryo: from early information to higher-level integration

Abstract: Although vertebrates seem to be essentially bilaterally symmetrical on the exterior, there are numerous interior left–right asymmetries in the disposition and placement of internal organs.

Abstract: These asymmetries are established during embryogenesis by complex epigenetic and genetic cascades.

Abstract: Recent studies in a range of model organisms have made important progress in understanding how this laterality information is generated and conveyed to large regions of the embryo.

Abstract: Both commonalities and divergences are emerging in the mechanisms that different vertebrates use in left–right axis specification.

Abstract: Recent evidence also provides intriguing links between the establishment of left–right asymmetries and the symmetrical elongation of the anterior–posterior axis.


Obesity, type 2 diabetes, and cancer: the insulin and IGF connection.

Abstract: Epidemiological studies suggest a positive association between obesity and type 2 diabetes mellitus (T2D) with the risk of cancer and cancer-related mortality.

Abstract: Insulin resistance, hyperinsulinemia, increased levels of IGF, elevated levels of steroid and peptide hormones, and inflammatory markers appear to play a role in the connection between these different diseases.

Abstract: Medications, such as metformin and exogenous insulin, used to treat T2D may affect the risk of cancer and cancer-related mortality.

Abstract: Newer therapies targeting the insulin and IGF1 systems are being developed for use in cancer therapy.


Non-coding RNAs: Functions and applications in endocrine-related cancer.

Abstract: A significant fraction of the human genome is transcribed as non-coding RNAs (ncRNAs).

Abstract: This non-coding transcriptome has challenged the notion of the central dogma and its involvement in transcriptional and post-transcriptional regulation of gene expression is well established.

Abstract: Interestingly, several ncRNAs are dysregulated in cancer and current non-coding transcriptome research aims to use our increasing knowledge of these ncRNAs for the development of cancer biomarkers and anti-cancer drugs.

Abstract: In endocrine-related cancers, for which survival rates can be relatively low, there is a need for such advancements.

Abstract: In this review, we aimed to summarize the roles and clinical implications of recently discovered ncRNAs, including long ncRNAs, PIWI-interacting RNAs, tRNA- and Y RNA-derived ncRNAs, and small nucleolar RNAs, in endocrine-related cancers affecting both sexes.

Abstract: We focus on recent studies highlighting discoveries in ncRNA biology and expression in cancer, and conclude with a discussion on the challenges and future directions, including clinical application.

Abstract: ncRNAs show great promise as diagnostic tools and therapeutic targets, but further work is necessary to realize the potential of these unconventional transcripts.


NMR structural and kinetic characterization of a homeodomain diffusing and hopping on nonspecific DNA.

Abstract: Nonspecific protein-DNA interactions are inherently dynamic and involve both diffusion of the protein along the DNA and hopping of the protein from one DNA molecule or segment to another.

Abstract: Understanding how gene regulatory proteins interact nonspecifically with DNA in terms of both structure and dynamics is challenging because the experimental observables are an ensemble average of many rapidly exchanging states.

Abstract: By using a variety of NMR spectroscopic techniques, including relaxation analysis, paramagnetic relaxation enhancement, and residual dipolar couplings, we have characterized structural and kinetic aspects of the interaction of the HoxD9 homeodomain with a nonspecific, 24-bp DNA duplex in a system in which the protein is not constrained to any particular site.

Abstract: The data reveal that HoxD9 binds to nonspecific DNA with the same binding mode and orientation as that observed in the specific complex.

Abstract: The mobility, however, of Arg side-chains contacting the DNA is increased in the nonspecific complex relative to the specific one.

Abstract: The kinetics of intermolecular translocation between two different nonspecific DNA molecules have also been analyzed and reveal that at high DNA concentrations (such as those present in vivo) direct transfer from one nonspecific complex to another nonspecific DNA molecule occurs without going through the intermediary of free protein.

Abstract: This finding provides a simple mechanism for accelerating the target search in vivo for the specific site in a sea of nonspecific sites by permitting more effective sampling of available DNA sites as the protein jumps from one segment to another.


MHC Haplotype Matching for Unrelated Hematopoietic Cell Transplantation

Abstract: Background  Current criteria for the selection of unrelated donors for hematopoietic cell transplantation (HCT) include matching for the alleles of each human leukocyte antigen (HLA) locus within the major histocompatibility complex (MHC).

Abstract: Graft-versus-host disease (GVHD), however, remains a significant and potentially life-threatening complication even after HLA-identical unrelated HCT.

Abstract: The MHC harbors more than 400 genes, but the total number of transplantation antigens is unknown.

Abstract: Genes that influence transplantation outcome could be identified by using linkage disequilibrium (LD)-mapping approaches, if the extended MHC haplotypes of the unrelated donor and recipient could be defined.


The SIR2/3/4 complex and SIR2 alone promote longevity in Saccharomyces cerevisiae by two different mechanisms. Genes Dev 13

Abstract: The SIR genes are determinants of life span in yeast mother cells.

Abstract: Here we show that life span regulation by the Sir proteins is independent of their role in nonhomologous end joining.

Abstract: The short life span of a sir3 or sir4 mutant is due to the simultaneous expression of a and alpha mating-type information, which indirectly causes an increase in rDNA recombination and likely increases the production of extrachromosomal rDNA circles.

Abstract: The short life span of a sir2 mutant also reveals a direct failure to repress recombination generated by the Fob1p-mediated replication block in the rDNA.

Abstract: Sir2p is a limiting component in promoting yeast longevity, and increasing the gene dosage extends the life span in wild-type cells.

Abstract: A possible role of the conserved SIR2 in mammalian aging is discussed.


Meta-analysis identifies common variants associated with body mass index in East Asians

Abstract: Multiple genetic loci associated with obesity or body mass index (BMI) have been identified through genome-wide association studies conducted predominantly in populations of European ancestry.

Abstract: We performed a meta-analysis of associations between BMI and approximately 2.4 million SNPs in 27,715 east Asians, which was followed by in silico and de novo replication studies in 37,691 and 17,642 additional east Asians, respectively.

Abstract: We identified ten BMI-associated loci at genome-wide significance (P < 5.0 × 10(-8)), including seven previously identified loci (FTO, SEC16B, MC4R, GIPR-QPCTL, ADCY3-DNAJC27, BDNF and MAP2K5) and three novel loci in or near the CDKAL1, PCSK1 and GP2 genes.

Abstract: Three additional loci nearly reached the genome-wide significance threshold, including two previously identified loci in the GNPDA2 and TFAP2B genes and a newly identified signal near PAX6, all of which were associated with BMI with P < 5.0 × 10(-7).

Abstract: Findings from this study may shed light on new pathways involved in obesity and demonstrate the value of conducting genetic studies in non-European populations.


Phosphorylation of histone H3(T118) alters nucleosome dynamics and remodeling

Abstract: Nucleosomes, the fundamental units of chromatin structure, are regulators and barriers to transcription, replication and repair.

Abstract: Post-translational modifications (PTMs) of the histone proteins within nucleosomes regulate these DNA processes.

Abstract: Histone H3(T118) is a site of phosphorylation [H3(T118ph)] and is implicated in regulation of transcription and DNA repair.

Abstract: We prepared H3(T118ph) by expressed protein ligation and determined its influence on nucleosome dynamics.

Abstract: We find H3(T118ph) reduces DNA-histone binding by 2  kcal/mol, increases nucleosome mobility by 28-fold and increases DNA accessibility near the dyad region by 6-fold.

Abstract: Moreover, H3(T118ph) increases the rate of hMSH2-hMSH6 nucleosome disassembly and enables nucleosome disassembly by the SWI/SNF chromatin remodeler.

Abstract: These studies suggest that H3(T118ph) directly enhances and may reprogram chromatin remodeling reactions.


Mechanosignaling through YAP and TAZ drives fibroblast activation and fibrosis.

Abstract: Pathological fibrosis is driven by a feedback loop in which the fibrotic extracellular matrix is both a cause and consequence of fibroblast activation.

Abstract: However, the molecular mechanisms underlying this process remain poorly understood.

Abstract: Here we identify yes-associated protein (YAP) (homolog of drosophila Yki) and transcriptional coactivator with PDZ-binding motif (TAZ) (also known as Wwtr1), transcriptional effectors of the Hippo pathway, as key matrix stiffness-regulated coordinators of fibroblast activation and matrix synthesis.

Abstract: YAP and TAZ are prominently expressed in fibrotic but not healthy lung tissue, with particularly pronounced nuclear expression of TAZ in spindle-shaped fibroblastic cells.

Abstract: In culture, both YAP and TAZ accumulate in the nuclei of fibroblasts grown on pathologically stiff matrices but not physiologically compliant matrices.

Abstract: Knockdown of YAP and TAZ together in vitro attenuates key fibroblast functions, including matrix synthesis, contraction, and proliferation, and does so exclusively on pathologically stiff matrices.

Abstract: Profibrotic effects of YAP and TAZ operate, in part, through their transcriptional target plasminogen activator inhibitor-1, which is regulated by matrix stiffness independent of transforming growth factor-β signaling.

Abstract: Immortalized fibroblasts conditionally expressing active YAP or TAZ mutant proteins overcome soft matrix limitations on growth and promote fibrosis when adoptively transferred to the murine lung, demonstrating the ability of fibroblast YAP/TAZ activation to drive a profibrotic response in vivo.

Abstract: Together, these results identify YAP and TAZ as mechanoactivated coordinators of the matrix-driven feedback loop that amplifies and sustains fibrosis.


Control of cellular cholesterol efflux by the nuclear oxysterol receptor LXR alpha.

Abstract: LXR alpha is a nuclear receptor that has previously been shown to regulate the metabolic conversion of cholesterol to bile acids.

Abstract: Here we define a role for this transcription factor in the control of cellular cholesterol efflux.

Abstract: We demonstrate that retroviral expression of LXR alpha in NIH 3T3 fibroblasts or RAW264.7 macrophages and/or treatment of these cells with oxysterol ligands of LXR results in 7- to 30-fold induction of the mRNA encoding the putative cholesterol/phospholipid transporter ATP-binding cassette (ABC)A1.

Abstract: In contrast, induction of ABCA1 mRNA in response to oxysterols is attenuated in cells that constitutively express dominant-negative forms of LXR alpha or LXR beta that lack the AF2 transcriptional activation domain.

Abstract: We further demonstrate that expression of LXR alpha in NIH 3T3 fibroblasts and/or treatment of these cells with oxysterols is sufficient to stimulate cholesterol efflux to extracellular apolipoprotein AI.

Abstract: The ability of oxysterol ligands of LXR to stimulate efflux is dramatically reduced in Tangier fibroblasts, which carry a loss of function mutation in the ABCA1 gene.

Abstract: Taken together, these results indicate that cellular cholesterol efflux is controlled, at least in part, at the level of transcription by a nuclear receptor-signaling pathway.

Abstract: They suggest a model in which activation of LXRs by oxysterols in response to cellular sterol loading leads to induction of the ABCA1 transporter and the stimulation of lipid efflux to extracellular acceptors.

Abstract: These findings have important implications for our understanding of mammalian cholesterol homeostasis and suggest new opportunities for pharmacological regulation of cellular lipid metabolism.


Circadian regulation of intracellular G-protein signalling mediates intercellular synchrony and rhythmicity in the suprachiasmatic nucleus

Abstract: Synchronous oscillations of thousands of cellular clocks in the suprachiasmatic nucleus (SCN), the circadian centre, are coordinated by precisely timed cell-cell communication, the principle of which is largely unknown.

Abstract: Here we show that the amount of RGS16 (regulator of G protein signalling 16), a protein known to inactivate Gαi, increases at a selective circadian time to allow time-dependent activation of intracellular cyclic AMP signalling in the SCN.

Abstract: Gene ablation of Rgs16 leads to the loss of circadian production of cAMP and as a result lengthens circadian period of behavioural rhythm.

Abstract: The temporally precise regulation of the cAMP signal by clock-controlled RGS16 is needed for the dorsomedial SCN to maintain a normal phase-relationship to the ventrolateral SCN.

Abstract: Thus, RGS16-dependent temporal regulation of intracellular G protein signalling coordinates the intercellular synchrony of SCN pacemaker neurons and thereby defines the 24 h rhythm in behaviour.