An actomyosin-based barrier inhibits cell mixing at compartmental boundaries in Drosophila embryos

Abstract: Partitioning tissues into compartments that do not intermix is essential for the correct morphogenesis of animal embryos and organs.

Abstract: Several hypotheses have been proposed to explain compartmental cell sorting, mainly differential adhesion, but also regulation of the cytoskeleton or of cell proliferation.

Abstract: Nevertheless, the molecular and cellular mechanisms that keep cells apart at boundaries remain unclear.

Abstract: Here we demonstrate, in early Drosophila melanogaster embryos, that actomyosin-based barriers stop cells from invading neighbouring compartments.

Abstract: Our analysis shows that cells can transiently invade neighbouring compartments, especially when they divide, but are then pushed back into their compartment of origin.

Abstract: Actomyosin cytoskeletal components are enriched at compartmental boundaries, forming cable-like structures when the epidermis is mitotically active.

Abstract: When MyoII (non-muscle myosin II) function is inhibited, including locally at the cable by chromophore-assisted laser inactivation (CALI), in live embryos, dividing cells are no longer pushed back, leading to compartmental cell mixing.

Abstract: We propose that local regulation of actomyosin contractibility, rather than differential adhesion, is the primary mechanism sorting cells at compartmental boundaries.


Clinical algorithms for malaria diagnosis lack utility among people of different age groups.

Abstract: We conducted a study to determine whether clinical algorithms would be useful in malaria diagnosis among people living in an area of moderate malaria transmission within Kilifi District in Kenya.

Abstract: A total of 1602 people of all age groups participated.

Abstract: We took smears and recorded clinical signs and symptoms (prompted or spontaneous) of all those presenting to the study clinic with a history of fever.

Abstract: A malaria case was defined as a person presenting to the clinic with a history of fever and concurrent parasitaemia.

Abstract: A set of clinical signs and symptoms (algorithms) with the highest sensitivity and specificity for diagnosing a malaria case was selected for the age groups </=5 years, 6-14 years and >/=15 years.

Abstract: These age-optimized derived algorithms were able to identify about 66% of the cases among those <15 years of age but only 23% of cases among adults.

Abstract: Were these algorithms to be used as a basis for a decision on treatment among those presenting to the clinic, 16% of children </=5 years, 44% of those 6-14 years of age and 66% of the adults who had a history of fever and parasitaemia >/=5000 parasites/microl of blood would be sent home without treatment.

Abstract: Clinical algorithms therefore appear to have little utility in malaria diagnosis, performing even worse in the older age groups, where avoiding unnecessary use of anti-malarials would make more drugs available to the really needy population of children under 5 years of age.


Large-scale genetic fine mapping and genotype-phenotype associations implicate polymorphism in the IL2RA region in type 1 diabetes

Abstract: Genome-wide association studies are now identifying disease-associated chromosome regions.

Abstract: However, even after convincing replication, the localization of the causal variant(s) requires comprehensive resequencing, extensive genotyping and statistical analyses in large sample sets leading to targeted functional studies.

Abstract: Here, we have localized the type 1 diabetes (T1D) association in the interleukin 2 receptor alpha (IL2RA) gene region to two independent groups of SNPs, spanning overlapping regions of 14 and 40 kb, encompassing IL2RA intron 1 and the 5′ regions of IL2RA and RBM17 (odds ratio = 2.04, 95% confidence interval = 1.70–2.45; P = 1.92 × 10−28; control frequency = 0.635).

Abstract: Furthermore, we have associated IL2RA T1D susceptibility genotypes with lower circulating levels of the biomarker, soluble IL-2RA (P = 6.28 × 10−28), suggesting that an inherited lower immune responsiveness predisposes to T1D.


Maternal obesity epigenetically alters visceral fat progenitor cell properties in male offspring mice.

Abstract: KEY POINTS Maternal obesity reduces adipogenic progenitor density in offspring adipose tissue.

Abstract: The ability of adipose tissue expansion in the offspring of obese mothers is limited and is associated with metabolic dysfunction of adipose tissue when challenged with a high-fat diet.

Abstract: Maternal obesity induces DNA demethylation in the promoter of zinc finger protein 423, which renders progenitor cells with a high adipogenic capacity.

Abstract: Maternal obesity demonstrates long-term effects on the adipogenic capacity of progenitor cells in offspring adipose tissue, demonstrating a developmental programming effect.

Abstract: ABSTRACT Maternal obesity (MO) programs offspring obesity and metabolic disorders, although the underlying mechanisms remain poorly defined.

Abstract: Progenitor cells are the source of new adipocytes.

Abstract: The present study aimed to test whether MO epigenetically predisposes adipocyte progenitors in the fat of offspring to adipogenic differentiation and subsequent depletion, which leads to a failure of adipose tissue plasticity under positive energy balance, contributing to adipose tissue metabolic dysfunction.

Abstract: C57BL/6 female mice were fed either a control diet (10% energy from fat) or a high-fat diet (45% energy from fat) for 8 weeks before mating.

Abstract: Male offspring of control (Con) and obese (OB) dams were weaned onto a regular (Reg) or obesogenic (Obe) diet until 3 months of age.

Abstract: At weaning, male OB offspring had a higher expression of Zinc finger protein 423 (zfp423), a key transcription factor in adipogenesis, as well as lower DNA methylation of its promoter in progenitors of epididymal fat compared to Con offspring, which was correlated with enhanced adipogenic differentiation.

Abstract: At 3 months of age, progenitor density was 30.9 ± 9.7% lower in OB/Obe compared to Con/Obe mice, accompanied by a limited expansion of the adipocyte number when challenged with a high-energy diet.

Abstract: This difference was associated with lower DNA methylation in the zfp423 promoter in the epididymal fat of OB/Obe offspring, which was correlated with greater macrophage chemotactic protein-1 and hypoxia-inducible factor 1α expression.

Abstract: In summary, MO epigenetically limits the expansion capacity of offspring adipose tissue, providing an explanation for the adipose metabolic dysfunction of male offspring in the setting of MO.


Quality Testing of Artemisinin-Based Antimalarial Drugs in Myanmar.

Abstract: Artemisinin-based combination therapies are the frontline treatment of Plasmodium falciparum malaria.

Abstract: The circulation of falsified and substandard artemisinin-based antimalarials in Southeast Asia has been a major predicament for the malaria elimination campaign.

Abstract: To provide an update of this situation, we purchased 153 artemisinin-containing antimalarials, as convenience samples, in private drug stores from different regions of Myanmar.

Abstract: The quality of these drugs in terms of their artemisinin derivative content was tested using specific dipsticks for these artemisinin derivatives, as point-of-care devices.

Abstract: A subset of these samples was further tested by high-performance liquid chromatography (HPLC).

Abstract: This survey identified that > 35% of the collected drugs were oral artesunate and artemether monotherapies.

Abstract: When tested with the dipsticks, all but one sample passed the assays, indicating that the detected artemisinin derivative content corresponded approximately to the labeled contents.

Abstract: However, one artesunate injection sample was found to contain no active ingredient at all by the dipstick assay and subsequent HPLC analysis.

Abstract: The continued circulation of oral monotherapies and the description, for the first time, of falsified parenteral artesunate provides a worrisome picture of the antimalarial drug quality in Myanmar during the malaria elimination phase, a situation that deserves more oversight from regulatory authorities.


LINE-1 Retrotransposition Activity in Human Genomes

Abstract: Highly active (i.e., "hot") long interspersed element-1 (LINE-1 or L1) sequences comprise the bulk of retrotransposition activity in the human genome; however, the abundance of hot L1s in the human population remains largely unexplored.

Abstract: Here, we used a fosmid-based, paired-end DNA sequencing strategy to identify 68 full-length L1s that are differentially present among individuals but are absent from the human genome reference sequence.

Abstract: The majority of these L1s were highly active in a cultured cell retrotransposition assay.

Abstract: Genotyping 26 elements revealed that two L1s are only found in Africa and that two more are absent from the H952 subset of the Human Genome Diversity Panel.

Abstract: Therefore, these results suggest that hot L1s are more abundant in the human population than previously appreciated, and that ongoing L1 retrotransposition continues to be a major source of interindividual genetic variation.


Immune cell promotion of metastasis

Abstract: Metastatic disease is the major cause of death from cancer, and immunotherapy and chemotherapy have had limited success in reversing its progression.

Abstract: Data from mouse models suggest that the recruitment of immunosuppressive cells to tumours protects metastatic cancer cells from surveillance by killer cells, which nullifies the effects of immunotherapy and thus establishes metastasis.

Abstract: Furthermore, in most cases, tumour-infiltrating immune cells differentiate into cells that promote each step of the metastatic cascade and thus are novel targets for therapy.

Abstract: In this Review, we describe how tumour-infiltrating immune cells contribute to the metastatic cascade and we discuss potential therapeutic strategies to target these cells.


PICH: a DNA translocase specially adapted for processing anaphase bridge DNA.

Abstract: The Plk1-interacting checkpoint helicase (PICH) protein localizes to ultrafine anaphase bridges (UFBs) in mitosis alongside a complex of DNA repair proteins, including the Bloom's syndrome protein (BLM).

Abstract: However, very little is known about the function of PICH or how it is recruited to UFBs.

Abstract: Using a combination of microfluidics, fluorescence microscopy, and optical tweezers, we have defined the properties of PICH in an in vitro model of an anaphase bridge.

Abstract: We show that PICH binds with a remarkably high affinity to duplex DNA, resulting in ATP-dependent protein translocation and extension of the DNA.

Abstract: Most strikingly, the affinity of PICH for binding DNA increases with tension-induced DNA stretching, which mimics the effect of the mitotic spindle on a UFB.

Abstract: PICH binding also appears to diminish force-induced DNA melting.

Abstract: We propose a model in which PICH recognizes and stabilizes DNA under tension during anaphase, thereby facilitating the resolution of entangled sister chromatids.


The Dynamics of Genome-wide DNA Methylation Reprogramming in Mouse Primordial Germ Cells

Abstract: Genome-wide DNA methylation reprogramming occurs in mouse primordial germ cells (PGCs) and preimplantation embryos, but the precise dynamics and biological outcomes are largely unknown.

Abstract: We have carried out whole-genome bisulfite sequencing (BS-Seq) and RNA-Seq across key stages from E6.5 epiblast to E16.5 PGCs.

Abstract: Global loss of methylation takes place during PGC expansion and migration with evidence for passive demethylation, but sequences that carry long-term epigenetic memory (imprints, CpG islands on the X chromosome, germline-specific genes) only become demethylated upon entry of PGCs into the gonads.

Abstract: The transcriptional profile of PGCs is tightly controlled despite global hypomethylation, with transient expression of the pluripotency network, suggesting that reprogramming and pluripotency are inextricably linked.

Abstract: Our results provide a framework for the understanding of the epigenetic ground state of pluripotency in the germline.


Major histocompatibility complex associations of ankylosing spondylitis are complex and involve further epistasis with ERAP1

Abstract: Ankylosing spondylitis (AS) is a common, highly heritable, inflammatory arthritis for which HLA-B*27 is the major genetic risk factor, although its role in the aetiology of AS remains elusive.

Abstract: To better understand the genetic basis of the MHC susceptibility loci, we genotyped 7,264 MHC SNPs in 22,647 AS cases and controls of European descent.

Abstract: We impute SNPs, classical HLA alleles and amino-acid residues within HLA proteins, and tested these for association to AS status.

Abstract: Here we show that in addition to effects due to HLA-B*27 alleles, several other HLA-B alleles also affect susceptibility.

Abstract: After controlling for the associated haplotypes in HLA-B, we observe independent associations with variants in the HLA-A, HLA-DPB1 and HLA-DRB1 loci.

Abstract: We also demonstrate that the ERAP1 SNP rs30187 association is not restricted only to carriers of HLA-B*27 but also found in HLA-B*40:01 carriers independently of HLA-B*27 genotype.


RNA sequencing identifies dysregulation of the human pancreatic islet transcriptome by the saturated fatty acid palmitate.

Abstract: Pancreatic β-cell dysfunction and death are central in the pathogenesis of type 2 diabetes (T2D).

Abstract: Saturated fatty acids cause β-cell failure and contribute to diabetes development in genetically predisposed individuals.

Abstract: Here we used RNA sequencing to map transcripts expressed in five palmitate-treated human islet preparations, observing 1,325 modified genes.

Abstract: Palmitate induced fatty acid metabolism and endoplasmic reticulum (ER) stress.

Abstract: Functional studies identified novel mediators of adaptive ER stress signaling.

Abstract: Palmitate modified genes regulating ubiquitin and proteasome function, autophagy, and apoptosis.

Abstract: Inhibition of autophagic flux and lysosome function contributed to lipotoxicity.

Abstract: Palmitate inhibited transcription factors controlling β-cell phenotype, including PAX4 and GATA6.

Abstract: Fifty-nine T2D candidate genes were expressed in human islets, and 11 were modified by palmitate.

Abstract: Palmitate modified expression of 17 splicing factors and shifted alternative splicing of 3,525 transcripts.

Abstract: Ingenuity Pathway Analysis of modified transcripts and genes confirmed that top changed functions related to cell death.

Abstract: Database for Annotation, Visualization and Integrated Discovery (DAVID) analysis of transcription factor binding sites in palmitate-modified transcripts revealed a role for PAX4, GATA, and the ER stress response regulators XBP1 and ATF6.

Abstract: This human islet transcriptome study identified novel mechanisms of palmitate-induced β-cell dysfunction and death.

Abstract: The data point to cross talk between metabolic stress and candidate genes at the β-cell level.


The Ratio of Monocytes to Lymphocytes in Peripheral Blood Correlates with Increased Susceptibility to Clinical Malaria in Kenyan Children

Abstract: BACKGROUND Plasmodium falciparum malaria remains a major cause of illness and death in sub-Saharan Africa.

Abstract: Young children bear the brunt of the disease and though older children and adults suffer relatively fewer clinical attacks, they remain susceptible to asymptomatic P. falciparum infection.

Abstract: A better understanding of the host factors associated with immunity to clinical malaria and the ability to sustain asymptomatic P. falciparum infection will aid the development of improved strategies for disease prevention.

Abstract: METHODS AND FINDINGS Here we investigate whether full differential blood counts can predict susceptibility to clinical malaria among Kenyan children sampled at five annual cross-sectional surveys.

Abstract: We find that the ratio of monocytes to lymphocytes, measured in peripheral blood at the time of survey, directly correlates with risk of clinical malaria during follow-up.

Abstract: This association is evident among children with asymptomatic P. falciparum infection at the time the cell counts are measured (Hazard ratio (HR)  =  2.7 (95% CI 1.42, 5.01, P  =  0.002) but not in those without detectable parasitaemia (HR  =  1.0 (95% CI 0.74, 1.42, P  =  0.9).

Abstract: CONCLUSIONS We propose that the monocyte to lymphocyte ratio, which is easily derived from routine full differential blood counts, reflects an individual's capacity to mount an effective immune response to P. falciparum infection.


Dynamic analysis of filopodial interactions during the zippering phase of Drosophila dorsal closure.

Abstract: Dorsal closure is a paradigm epithelial fusion episode that occurs late in Drosophila embryogenesis and leads to sealing of a midline hole by bonding of two opposing epithelial sheets.

Abstract: The leading edge epithelial cells express filopodia and fusion is dependent on interdigitation of these filopodia to prime formation of adhesions.

Abstract: Since the opposing epithelia are molecularly patterned there must exist some mechanism for accurately aligning the two sheets across this fusion seam.

Abstract: To address this, we generated a fly in which RFP-Moesin and GFP-Moesin are expressed in mutually exclusive stripes within each segment using the engrailed and patched promoters.

Abstract: We observe mutually exclusive interactions between the filopodia of engrailed and patched cells.

Abstract: Interactions between filopodia from matching cells leads to formation of tethers between them, and these tethers can pull misaligned epithelial sheets into alignment.

Abstract: Filopodial matching also occurs during repair of laser wounds in the ventral epithelium, and so this behaviour is not restricted to leading edge cells during dorsal closure.

Abstract: Finally, we characterise the behaviour of a patched-expressing cell that we observe within the engrailed region of segments A1-A5, and provide evidence that this cell contributes to cell matching.


Expression of Versican 3′-Untranslated Region Modulates Endogenous MicroRNA Functions

Abstract: BACKGROUND Mature microRNAs (miRNAs) are single-stranded RNAs that regulate post-transcriptional gene expression.

Abstract: In our previous study, we have shown that versican 3'UTR, a fragment of non-coding transcript, has the ability to antagonize miR-199a-3p function thereby regulating expression of the matrix proteins versican and fibronectin, and thus resulting in enhanced cell-cell adhesion and organ adhesion.

Abstract: However, the impact of this non-coding fragment on tumorigenesis is yet to be determined.

Abstract: METHODS AND FINDINGS Using computational prediction confirmed with in vitro and in vivo experiments, we report that the expression of versican 3'UTR not only antagonizes miR-199a-3p but can also lower its steady state expression.

Abstract: We found that expression of versican 3'UTR in a mouse breast carcinoma cell line, 4T1, decreased miR-199a-3p levels.

Abstract: The decrease in miRNA activity consequently translated into differences in tumor growth.

Abstract: Computational analysis indicated that both miR-199a-3p and miR-144 targeted a cell cycle regulator, Rb1.

Abstract: In addition, miR-144 and miR-136, which have also been shown to interact with versican 3'UTR, was found to target PTEN.

Abstract: Expression of Rb1 and PTEN were up-regulated synergistically in vitro and in vivo, suggesting that the 3'UTR binds and modulates miRNA activities, freeing Rb1 and PTEN mRNAs for translation.

Abstract: In tumor formation assays, cells transfected with the 3'UTR formed smaller tumors compared with cells transfected with a control vector.

Abstract: CONCLUSION Our results demonstrated that a 3'UTR fragment can be used to modulate miRNA functions.

Abstract: Our study also suggests that miRNAs in the cancer cells are more susceptible to degradation, due to its interaction with a non-coding 3'UTR.

Abstract: This non-coding component of mRNA may be used retrospectively to modulate miRNA activities.


Prophylactic and therapeutic efficacy of the epitope vaccine CTB-UA against Helicobacter pylori infection in a BALB/c mice model

Abstract: Epitope vaccine based on the enzyme urease of Helicobacter pylori is a promising option for prophylactic and therapeutic vaccination against H. pylori infection.

Abstract: In our previous study, the epitope vaccine CTB-UA, which was composed of the mucosal adjuvant cholera toxin B subunit (CTB) and an epitope (UreA183–203) from the H. pylori urease A subunit (UreA) was constructed.

Abstract: This particular vaccine was shown to have good immunogenicity and immunoreactivity and could induce specific neutralizing antibodies, which exhibited effectively inhibitory effects on the enzymatic activity of H. pylori urease.

Abstract: In this study, the prophylactic and therapeutic efficacy of the epitope vaccine CTB-UA was evaluated in a BALB/c mice model.

Abstract: The experimental results indicated that oral prophylactic or therapeutic immunization with CTB-UA significantly decreased H. pylori colonization compared with oral immunization with PBS.

Abstract: The results also revealed that the protection was correlated with antigen-specific IgG, IgA, and mucosal secretory IgA antibody responses.

Abstract: CTB-UA may be a promising vaccine candidate for the control of H. pylori infection.


Spatial co-distribution of neglected tropical diseases in the east African great lakes region: revisiting the justification for integrated control.

Abstract: OBJECTIVE To determine spatial patterns of co-endemicity of schistosomiasis mansoni and the soil-transmitted helminths (STHs) Ascaris lumbricoides, Trichuris trichiura and hookworm in the Great Lakes region of East Africa, to help plan integrated neglected tropical disease programmes in this region.

Abstract: METHOD Parasitological surveys were conducted in Uganda, Tanzania, Kenya and Burundi in 28 213 children in 404 schools.

Abstract: Bayesian geostatistical models were used to interpolate prevalence of these infections across the study area.

Abstract: Interpolated prevalence maps were overlaid to determine areas of co-endemicity.

Abstract: RESULTS In the Great Lakes region, prevalence was 18.1% for Schistosoma mansoni, 50.0% for hookworm, 6.8% for A. lumbricoides and 6.8% for T. trichiura.

Abstract: Hookworm infection was ubiquitous, whereas S. mansoni, A. lumbricoides and T. trichiura were highly focal.

Abstract: Most areas were endemic (prevalence >or=10%) or hyperendemic (prevalence >or=50%) for one or more STHs, whereas endemic areas for schistosomiasis mansoni were restricted to foci adjacent large perennial water bodies.

Abstract: CONCLUSION Because of the ubiquity of hookworm, treatment programmes are required for STH throughout the region but efficient schistosomiasis control should only be targeted at limited high-risk areas.

Abstract: Therefore, integration of schistosomiasis with STH control is only indicated in limited foci in East Africa.


Direct conversion of patient fibroblasts demonstrates non-cell autonomous toxicity of astrocytes to motor neurons in familial and sporadic ALS.

Abstract: Amyotrophic lateral sclerosis (ALS) causes motor neuron degeneration, paralysis, and death.

Abstract: Accurate disease modeling, identifying disease mechanisms, and developing therapeutics is urgently needed.

Abstract: We previously reported motor neuron toxicity through postmortem ALS spinal cord-derived astrocytes.

Abstract: However, these cells can only be harvested after death, and their expansion is limited.

Abstract: We now report a rapid, highly reproducible method to convert adult human fibroblasts from living ALS patients to induced neuronal progenitor cells and subsequent differentiation into astrocytes (i-astrocytes).

Abstract: Non-cell autonomous toxicity to motor neurons is found following coculture of i-astrocytes from familial ALS patients with mutation in superoxide dismutase or hexanucleotide expansion in C9orf72 (ORF 72 on chromosome 9) the two most frequent causes of ALS.

Abstract: Remarkably, i-astrocytes from sporadic ALS patients are as toxic as those with causative mutations, suggesting a common mechanism.

Abstract: Easy production and expansion of i-astrocytes now enables rapid disease modeling and high-throughput drug screening to alleviate astrocyte-derived toxicity.


Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations

Abstract: Heterogeneity within the self-renewal durability of adult hematopoietic stem cells (HSCs) challenges our understanding of the molecular framework underlying HSC function.

Abstract: Gene expression studies have been hampered by the presence of multiple HSC subtypes and contaminating non-HSCs in bulk HSC populations.

Abstract: To gain deeper insight into the gene expression program of murine HSCs, we combined single-cell functional assays with flow cytometric index sorting and single-cell gene expression assays.

Abstract: Through bioinformatic integration of these datasets, we designed an unbiased sorting strategy that separates non-HSCs away from HSCs, and single-cell transplantation experiments using the enriched population were combined with RNA-seq data to identify key molecules that associate with long-term durable self-renewal, producing a single-cell molecular dataset that is linked to functional stem cell activity.

Abstract: Finally, we demonstrated the broader applicability of this approach for linking key molecules with defined cellular functions in another stem cell system.


Acids: Structures, Properties, and Functions (University Science Books, Sausalito, CA, 2000).

Abstract: Humans expressing a defective form of the transcription factor AIRE (autoimmune regulator) develop multiorgan autoimmune disease.

Abstract: We used aire- deficient mice to test the hypothesis that this transcription factor regulates autoimmunity by promoting the ectopic expression of peripheral tissue- restricted antigens in medullary epithelial cells of the thymus.

Abstract: This hypothesis proved correct.

Abstract: The mutant animals exhibited a defined profile of autoimmune diseases that depended on the absence of aire in stromal cells of the thymus.

Abstract: Aire-deficient thymic medullary epithelial cells showed a specific reduction in ectopic transcription of genes encoding peripheral antigens.

Abstract: These findings highlight the importance of thymically imposed "central" tolerance in controlling autoimmunity.


Directed Actin Polymerization Is the Driving Force for Epithelial Cell–Cell Adhesion

Abstract: We have found that epithelial cells engage in a process of cadherin-mediated intercellular adhesion that utilizes calcium and actin polymerization in unexpected ways.

Abstract: Calcium stimulates filopodia, which penetrate and embed into neighboring cells.

Abstract: E-cadherin complexes cluster at filopodia tips, generating a two-rowed zipper of embedded puncta.

Abstract: Opposing cell surfaces are clamped by desmosomes, while vinculin, zyxin, VASP, and Mena are recruited to adhesion zippers by a mechanism that requires alpha-catenin.

Abstract: Actin reorganizes and polymerizes to merge puncta into a single row and seal cell borders.

Abstract: In keratinocytes either null for alpha-catenin or blocked in VASP/Mena function, filopodia embed, but actin reorganization/polymerization is prevented, and membranes cannot seal.

Abstract: Taken together, a dynamic mechanism for intercellular adhesion is unveiled involving calcium-activated filopodia penetration and VASP/Mena-dependent actin reorganization/polymerization.


Ribosomal DNA copy number loss and sequence variation in cancer

Abstract: Ribosomal DNA is one of the most variable regions in the human genome with respect to copy number.

Abstract: Despite the importance of rDNA for cellular function, we know virtually nothing about what governs its copy number, stability, and sequence in the mammalian genome due to challenges associated with mapping and analysis.

Abstract: We applied computational and droplet digital PCR approaches to measure rDNA copy number in normal and cancer states in human and mouse genomes.

Abstract: We find that copy number and sequence can change in cancer genomes.

Abstract: Counterintuitively, human cancer genomes show a loss of copies, accompanied by global copy number co-variation.

Abstract: The sequence can also be more variable in the cancer genome.

Abstract: Cancer genomes with lower copies have mutational evidence of mTOR hyperactivity.

Abstract: The PTEN phosphatase is a tumor suppressor that is critical for genome stability and a negative regulator of the mTOR kinase pathway.

Abstract: Surprisingly, but consistent with the human cancer genomes, hematopoietic cancer stem cells from a Pten-/- mouse model for leukemia have lower rDNA copy number than normal tissue, despite increased proliferation, rRNA production, and protein synthesis.

Abstract: Loss of copies occurs early and is associated with hypersensitivity to DNA damage.

Abstract: Therefore, copy loss is a recurrent feature in cancers associated with mTOR activation.

Abstract: Ribosomal DNA copy number may be a simple and useful indicator of whether a cancer will be sensitive to DNA damaging treatments.


The association of Red cell distribution width and in-hospital mortality in older adults admitted to the emergency department

Abstract: BACKGROUND The objective of the study was to test the hypothesis that elevated red cell distribution width (RDW) at admission increases the risk of mortality in older patients admitted to the emergency department (ED).

Abstract: METHODS We performed a retrospective analysis of patients admitted to the ED between May 2013 and October 2013.

Abstract: We included patients who were older than 65 years who visited the ED with any medical problems.

Abstract: Baseline RDW values were measured at the time of admission to the ED.

Abstract: The primary outcome was all-cause in-hospital mortality.

Abstract: Multivariate logistic analysis was performed.

Abstract: RESULTS A total of 1,990 patients were finally included in this study.

Abstract: The mean age was 75 years (SD 7), and 936 (47 %) subjects were male.

Abstract: The in-hospital mortality rate was 3.76 % (74 patients).

Abstract: RDW values higher in non-survivors than in survivors (15.9 ± 2.5 vs. 13.8 ± 1.7, p < 0.001).

Abstract: Multivariate logistic analysis showed that RDW was associated with all-cause in-hospital mortality after adjusting for other confounding factors.

Abstract: DISCUSSION RDW value at admission is an independent predictor of all-cause in-hospital mortality among patients older than 65 years.

Abstract: After adjustment for multiple confounders, the all-cause in-hospital mortality rate increased by 21.8% for each 1% increase in RDW.

Abstract: CONCLUSION These results show that RDW at admission is associated with in-hospital mortality among patients older than 65.

Abstract: Thus, RDW at admission may represent a surrogate marker of disease severity.

Abstract: We caution against using these findings to aid clinical decision-making process until they are externally validated.


Gene regulation by the act of long non-coding RNA transcription

Abstract: Long non-protein-coding RNAs (lncRNAs) are proposed to be the largest transcript class in the mouse and human transcriptomes.

Abstract: Two important questions are whether all lncRNAs are functional and how they could exert a function.

Abstract: Several lncRNAs have been shown to function through their product, but this is not the only possible mode of action.

Abstract: In this review we focus on a role for the process of lncRNA transcription, independent of the lncRNA product, in regulating protein-coding-gene activity in cis.

Abstract: We discuss examples where lncRNA transcription leads to gene silencing or activation, and describe strategies to determine if the lncRNA product or its transcription causes the regulatory effect.


Life and death of cardiac stem cells: a paradigm shift in cardiac biology.

Abstract: The recognition that myocyte mitosis occurs in the fetal, neonatal, adult, and hypertrophied heart and that a pool of primitive, undifferentiated cells is present in the myocardium has put forward a different view of the biology of the heart.

Abstract: The new paradigm suggests that myocyte formation is preserved during postnatal life, in adulthood or senescence, pointing to a remarkable growth reserve of the heart throughout the course of life of the organism.

Abstract: This article reviews a large body of novel information, which has been obtained in the last 2 decades, in favor of the notion that the mammalian heart has the inherent ability to continuously replace its parenchymal cells and that this unexpected characteristic has important implications in understanding myocardial homeostasis, cardiac aging, and tissue repair.

Abstract: The paradigm that the heart is a postmitotic organ incapable of regenerating parenchymal cells was established in the 1970s, and this dogma has profoundly conditioned basic and clinical research in cardiology for the last 3 decades.

Abstract: On the basis of this paradigm, cardiomyocytes undergo cellular hypertrophy1,2 but cannot be replaced either by entry into the cell cycle of a subpopulation of nonterminally differentiated myocytes or by activation of a pool of primitive cells that become committed to the myocyte lineage.

Abstract: The only response of cardiomyocytes to stress is hypertrophy and/or death.

Abstract: Therefore, a tremendous effort was made to identify the molecular mechanisms of myocyte hypertrophy and their genetic control.

Abstract: A sophisticated knowledge of various signaling pathways has been achieved, and our understanding of the biology of hypertrophic myocyte growth has advanced markedly.3 An array of new technologies has been introduced that has led to a scientific revolution in terms of questions, approaches, and interpretation of experimental results.

Abstract: Despite this enormous progress in our understanding of basic mechanisms of hypertrophy, however, very little …


Paraneoplastic thrombocytosis: the secrets of tumor self-promotion.

Abstract: Paraneoplastic thrombocytosis is associated with many solid tumors and often correlates with reduced survival.

Abstract: Recent studies suggest that a pathogenic feed back loop may be operative between platelets and tumor cells, with reciprocal interactions between tumor growth/metastasis and thrombocytosis/platelet activation.

Abstract: Specific molecular pathways have been identified in which tumors can stimulate platelet production and activation; activated platelets can, in turn, promote tumor growth and metastasis.

Abstract: Taken together, these findings provide exciting new potential targets for therapeutic intervention.


Perceived age as clinically useful biomarker of ageing: cohort study.

Abstract: OBJECTIVE To determine whether perceived age correlates with survival and important age related phenotypes.

Abstract: DESIGN Follow-up study, with survival of twins determined up to January 2008, by which time 675 (37%) had died.

Abstract: SETTING Population based twin cohort in Denmark.

Abstract: PARTICIPANTS 20 nurses, 10 young men, and 11 older women (assessors); 1826 twins aged >or=70.

Abstract: MAIN OUTCOME MEASURES Assessors: perceived age of twins from photographs.

Abstract: Twins: physical and cognitive tests and molecular biomarker of ageing (leucocyte telomere length).

Abstract: RESULTS For all three groups of assessors, perceived age was significantly associated with survival, even after adjustment for chronological age, sex, and rearing environment.

Abstract: Perceived age was still significantly associated with survival after further adjustment for physical and cognitive functioning.

Abstract: The likelihood that the older looking twin of the pair died first increased with increasing discordance in perceived age within the twin pair-that is, the bigger the difference in perceived age within the pair, the more likely that the older looking twin died first.

Abstract: Twin analyses suggested that common genetic factors influence both perceived age and survival.

Abstract: Perceived age, controlled for chronological age and sex, also correlated significantly with physical and cognitive functioning as well as with leucocyte telomere length.

Abstract: CONCLUSION Perceived age-which is widely used by clinicians as a general indication of a patient's health-is a robust biomarker of ageing that predicts survival among those aged >or=70 and correlates with important functional and molecular ageing phenotypes.


Inflammatory Reaction as Determinant of Foreign Body Reaction Is an Early and Susceptible Event after Mesh Implantation

Abstract: PURPOSE To investigate and relate the ultrashort-term and long-term courses of determinants for foreign body reaction as biocompatibility predictors for meshes in an animal model.

Abstract: MATERIALS AND METHODS Three different meshes (TVT, UltraPro, and PVDF) were implanted in sheep.

Abstract: Native and plasma coated meshes were placed bilaterally: (a) interaperitoneally, (b) as fascia onlay, and (c) as muscle onlay (fascia sublay).

Abstract: At 5 min, 20 min, 60 min, and 120 min meshes were explanted and histochemically investigated for inflammatory infiltrate, macrophage infiltration, vessel formation, myofibroblast invasion, and connective tissue accumulation.

Abstract: The results were related to long-term values over 24 months.

Abstract: RESULTS Macrophage invasion reached highest extents with up to 60% in short-term and decreased within 24 months to about 30%.

Abstract: Inflammatory infiltrate increased within the first 2 hours, the reached levels and the different extents and ranking among the investigated meshes remained stable during long-term follow up.

Abstract: For myofibroblasts, connective tissue, and CD31+ cells, no activity was detected during the first 120 min.

Abstract: CONCLUSION The local inflammatory reaction is an early and susceptible event after mesh implantation.

Abstract: It cannot be influenced by prior plasma coating and does not depend on the localisation of implantation.


Riesz pyramids for fast phase-based video magnification

Abstract: We present a new compact image pyramid representation, the Riesz pyramid, that can be used for real-time phase-based motion magnification.

Abstract: Our new representation is less overcomplete than even the smallest two orientation, octave-bandwidth complex steerable pyramid, and can be implemented using compact, efficient linear filters in the spatial domain.

Abstract: Motion-magnified videos produced with this new representation are of comparable quality to those produced with the complex steerable pyramid.

Abstract: When used with phase-based video magnification, the Riesz pyramid phase-shifts image features along only their dominant orientation rather than every orientation like the complex steerable pyramid.


Alpha-Synuclein Induces Lysosomal Rupture and Cathepsin Dependent Reactive Oxygen Species Following Endocytosis

Abstract: α-synuclein dysregulation is a critical aspect of Parkinson's disease pathology.

Abstract: Recent studies have observed that α-synuclein aggregates are cytotoxic to cells in culture and that this toxicity can be spread between cells.

Abstract: However, the molecular mechanisms governing this cytotoxicity and spread are poorly characterized.

Abstract: Recent studies of viruses and bacteria, which achieve their cytoplasmic entry by rupturing intracellular vesicles, have utilized the redistribution of galectin proteins as a tool to measure vesicle rupture by these organisms.

Abstract: Using this approach, we demonstrate that α-synuclein aggregates can induce the rupture of lysosomes following their endocytosis in neuronal cell lines.

Abstract: This rupture can be induced by the addition of α-synuclein aggregates directly into cells as well as by cell-to-cell transfer of α-synuclein.

Abstract: We also observe that lysosomal rupture by α-synuclein induces a cathepsin B dependent increase in reactive oxygen species (ROS) in target cells.

Abstract: Finally, we observe that α-synuclein aggregates can induce inflammasome activation in THP-1 cells.

Abstract: Lysosomal rupture is known to induce mitochondrial dysfunction and inflammation, both of which are well established aspects of Parkinson's disease, thus connecting these aspects of Parkinson's disease to the propagation of α-synuclein pathology in cells.


Mechanistic Fracture Criteria For The Failure Of Human Cortical Bone

Abstract: A mechanistic understanding of fracture in human bone is critical to predicting fracture risk associated with age and disease.

Abstract: Despite extensive work, a mechanistic framework for describing how the microstructure affects the failure of bone is lacking.

Abstract: Although micromechanical models incorporating local failure criteria have been developed for metallic and ceramic materials, few such models exist for biological materials.

Abstract: In fact, there is no proof to support the widely held belief that fracture in bone is locally strain-controlled, as for example has been shown for ductile fracture in metallic materials.

Abstract: In the present study, we provide such evidence through a novel series of experiments involving a double-notch-bend geometry, designed to shed light on the nature of the critical failure events in bone.

Abstract: We examine how the propagating crack interacts with the bone microstructure to provide some mechanistic understanding of fracture and to define how properties vary with orientation.

Abstract: It was found that fracture in human cortical bone is consistent with strain-controlled failure, and the influence of microstructure can be described in terms of several toughening mechanisms.

Abstract: We provide estimates of the relative importance of these mechanisms, such as uncracked-ligament bridging.


Inhibition of limb regeneration in the axolotl after treatment of the skin with actinomycin D.

Abstract: In this experiment actinomycin D was used to explore the action of the wound epidermis on underlying tissues during limb regeneration.

Abstract: In axolotl forelimbs the skin was removed from the elbow to the shoulder.

Abstract: Skin from the right limbs was soaked for three hours in actinomycin D (5.0 or 10.0 μg/ml 0.6% NaCl).

Abstract: For controls, skin from left limbs was soaked in 0.6% NaCl for the same period of time.

Abstract: Each piece of skin was orthotopically replanted, and both limbs were amputated through the treated skin, proximal to the elbow.

Abstract: After an initial healing period, the control limbs regenerated normally.

Abstract: Except for a slightly paler color, limbs bearing actinomycin-treated skin were indistinguishable from the controls, both grossly and histologically, during the first week following amputation.

Abstract: While the control limbs formed early blastemas, no grossly visible evidence of regeneration was apparent in the experimental limbs, but histologically some dedifferentiation was occurring.

Abstract: Normally three to four digits were seen in the control regenerates before blastemas appeared on the experimental limbs.

Abstract: By 35–40 days blastemas had appeared on most experimental limbs.

Abstract: These developed very rapidly, and within a short time many of them had attained levels of development close to the controls.

Abstract: Actinomycin D temporarily suppresses formation of the apical epidermal cap and the subsequent aggregation of dedifferentiated cells into a blastema.

Abstract: When the effect wears off, an apical cap forms and the dedifferentiated cells quickly organize into a blastema and begin to differentiate.


Retinoic acid-related orphan receptors α and γ: key regulators of lipid/glucose metabolism, inflammation, and insulin sensitivity

Abstract: Retinoic acid-related orphan receptors RORα and RORγ play a regulatory role in lipid/glucose homeostasis and various immune functions, and have been implicated in metabolic syndrome and several inflammatory diseases.

Abstract: RORα-deficient mice are protected against age- and diet-induced obesity, hepatosteatosis, and insulin resistance.

Abstract: The resistance to hepatosteatosis in RORα-deficient mice is related to the reduced expression of several genes regulating lipid synthesis, transport, and storage.

Abstract: Adipose tissue-associated inflammation, which plays a critical role in the development of insulin resistance, is considerably diminished in RORα-deficient mice as indicated by the reduced infiltration of M1 macrophages and decreased expression of many proinflammatory genes.

Abstract: Deficiency in RORγ also protects against diet-induced insulin resistance by a mechanism that appears different from that in RORα deficiency.

Abstract: Recent studies indicated that RORs provide an important link between the circadian clock machinery and its regulation of metabolic genes and metabolic syndrome.

Abstract: As ligand-dependent transcription factors, RORs may provide novel therapeutic targets in the management of obesity and associated metabolic diseases, including hepatosteatosis, adipose tissue-associated inflammation, and insulin resistance.


ERS/ESTS clinical guidelines on fitness for radical therapy in lung cancer patients (surgery and chemo-radiotherapy).

Abstract: A collaboration of multidisciplinary experts on the functional evaluation of lung cancer patients has been facilitated by the European Respiratory Society (ERS) and the European Society of Thoracic Surgery (ESTS), in order to draw up recommendations and provide clinicians with clear, up-to-date guidelines on fitness for surgery and chemo-radiotherapy.

Abstract: The subject was divided into different topics, which were then assigned to at least two experts.

Abstract: The authors searched the literature according to their own strategies, with no central literature review being performed.

Abstract: The draft reports written by the experts on each topic were reviewed, discussed and voted on by the entire expert panel.

Abstract: The evidence supporting each recommendation was summarised, and graded as described by the Scottish Intercollegiate Guidelines Network Grading Review Group.

Abstract: Clinical practice guidelines were generated and finalized in a functional algorithm for risk stratification of the lung resection candidates, emphasising cardiological evaluation, forced expiratory volume in 1 s, systematic carbon monoxide lung diffusion capacity and exercise testing.

Abstract: Contrary to lung resection, for which the scientific evidences are more robust, we were unable to recommend any specific test, cut-off value, or algorithm before chemo-radiotherapy due to the lack of data.

Abstract: We recommend that lung cancer patients should be managed in specialised settings by multidisciplinary teams.


The Ran GTPase regulates mitotic spindle assembly

Abstract: Ran is an abundant nuclear GTPase with a clear role in nuclear transport during interphase but with roles in mitotic regulation that are less well understood.

Abstract: The nucleotide-binding state of Ran is regulated by a GTPase activating protein, RanGAP1, and by a guanine nucleotide exchange factor, RCC1.

Abstract: Ran also interacts with a guanine nucleotide dissociation inhibitor, RanBP1.

Abstract: RanBP1 has a high affinity for GTP-bound Ran, and it acts as a cofactor for RanGAP1, increasing the rate of GAP-mediated GTP hydrolysis on Ran approximately tenfold.

Abstract: RanBP1 levels oscillate during the cell cycle [4], and increased concentrations of RanBP1 prolong mitosis in mammalian cells and in Xenopus egg extracts (our unpublished observations).

Abstract: We investigated how increased concentrations of RanBP1 disturb mitosis.

Abstract: We found that spindle assembly is dramatically disrupted when exogenous RanBP1 is added to M phase Xenopus egg extracts.

Abstract: We present evidence that the role of Ran in spindle assembly is independent of nuclear transport and is probably mediated through changes in microtubule dynamics.


A summary of the effects of antihypertensive medications on measured blood pressure.

Abstract: BACKGROUND Epidemiologic analysis of family data on blood pressure (BP) is often compromised by the effects of antihypertensive medications.

Abstract: A review of numerous clinical trials that investigated the effects of BP-lowering medications is summarized here.

Abstract: METHODS Published clinical trials, including 137 clinical trials with monodrug therapies and 28 clinical trials of combination drug therapies with a total of 11,739 participants, were reviewed from PubMed.

Abstract: Six major classes/groups of antihypertensive medications were categorized by ethnicity, including angiotensin-converting enzyme (ACE) inhibitors, alpha1-blockers, cardioselective beta-blockers (beta1-blockers), calcium channel blockers, thiazide and thiazide-like diuretics, and loop diuretics.

Abstract: RESULTS Using sitting or supine BP, for ethnic groups combined, monodrug therapy with ACE inhibitors showed a weighted average effect of lowering the systolic and diastolic BP by 12.5/9.5 mm Hg; alpha1-blockers by 15.5/11.7 mm Hg; beta1-blockers by 14.8/12.2 mm Hg; calcium channel blockers by 15.3/10.5 mm Hg; thiazide diuretics by 15.3/9.8 mm Hg; and loop diuretics by 15.8/8.2 mm Hg.

Abstract: However, ACE inhibitors, alpha1-blockers, and beta1-blockers were less effective in African Americans than in non-African Americans, whereas calcium channel blockers, thiazide diuretics, and loop diuretics were more effective in African Americans than in non-African Americans.

Abstract: For two-drug combination therapy with ethnic groups combined, the BP-lowering effect of the second medication, when compared to its effect as monodrug therapy, was 84% and 65% for systolic and diastolic BP, respectively.

Abstract: CONCLUSIONS The BP-lowering effects reported here may be used to impute the pretreatment BP levels, which can improve the information content and hence the power of epidemiologic analysis in studies where use of antihypertensive medications is a confounding factor in the BP measurements.


Stem Cells and Early Lineage Development

Abstract: The recent derivation of pluripotent stem cell lines from a number of different sources, including reprogrammed adult somatic cells, raises the issue of the developmental equivalence of these different pluripotent states.

Abstract: At least two different states representing the epiblast progenitors in the blastocyst and the pluripotent progenitors of the later gastrulating embryo have been recognized.

Abstract: Understanding the initial developmental status of the different pluripotent lines is critical for defining starting conditions for differentiation toward therapeutically relevant cell types.


Divergent clonal evolution of castration resistant neuroendocrine prostate cancer

Abstract: An increasingly recognized resistance mechanism to androgen receptor (AR)-directed therapy in prostate cancer involves epithelial plasticity, in which tumor cells demonstrate low to absent AR expression and often have neuroendocrine features.

Abstract: The etiology and molecular basis for this 'alternative' treatment-resistant cell state remain incompletely understood.

Abstract: Here, by analyzing whole-exome sequencing data of metastatic biopsies from patients, we observed substantial genomic overlap between castration-resistant tumors that were histologically characterized as prostate adenocarcinomas (CRPC-Adeno) and neuroendocrine prostate cancer (CRPC-NE); analysis of biopsy samples from the same individuals over time points to a model most consistent with divergent clonal evolution.

Abstract: Genome-wide DNA methylation analysis revealed marked epigenetic differences between CRPC-NE tumors and CRPC-Adeno, and also designated samples of CRPC-Adeno with clinical features of AR independence as CRPC-NE, suggesting that epigenetic modifiers may play a role in the induction and/or maintenance of this treatment-resistant state.

Abstract: This study supports the emergence of an alternative, 'AR-indifferent' cell state through divergent clonal evolution as a mechanism of treatment resistance in advanced prostate cancer.


S-phase-promoting cyclin-dependent kinases prevent re-replication by inhibiting the transition of replication origins to a pre-replicative state

Abstract: BACKGROUND DNA replication and mitosis are triggered by activation of kinase complexes, each made up of a cyclin and a cyclin-dependent kinase (Cdk).

Abstract: It had seemed possible that the association of Cdks with different classes of cyclins specifies whether S phase (replication) or M phase (mitosis) will occur.

Abstract: The recent finding that individual B-type cyclins (encoded by the genes CLB1-CLB6) can have functions in both processes in the budding yeast Saccharomyces cerevisiae casts doubt on this notion.

Abstract: RESULTS S. cerevisiae strains lacking C1b1-C1b4 undergo DNA replication once but fail to enter mitosis.

Abstract: We have isolated mutations in two genes, SIM1 and SIM2 (SIM2 is identical to SEC72), which allow such cells to undergo an extra round of DNA replication without mitosis.

Abstract: The Clb5 kinase, which promotes S phase, remains active during the G2-phase arrest of cells of the parental strain, but its activity declines rapidly in sim mutants.

Abstract: Increased expression of the CLB5 gene prevents re-replication.

Abstract: Thus, a cyclin B-kinase that promotes DNA replication in G1-phase cells can prevent re-replication in G2-phase cells.

Abstract: Inactivation of C1b kinases by expression of the specific C1b-Cdk1 inhibitor p40SIC1 is sufficient to induce a prereplicative state at origins of replication in cells blocked in G2/M phase by nocodazole.

Abstract: Re-activation of C1b-Cdk1 kinases induces a second round of DNA replication.

Abstract: CONCLUSIONS We propose that S-phase-promoting cyclin B--Cdk complexes prevent re-replication during S, G2 and M phases by inhibiting the transition of replication origins to a pre-replicative state.

Abstract: This model can explain both why origins 'fire' only once per S phase and why S phase is dependent on completion of the preceding M phase.


E-cadherin engagement stimulates proliferation via Rac1

Abstract: E-cadherin has been linked to the suppression of tumor growth and the inhibition of cell proliferation in culture.

Abstract: We observed that progressively decreasing the seeding density of normal rat kidney-52E (NRK-52E) or MCF-10A epithelial cells from confluence, indeed, released cells from growth arrest.

Abstract: Unexpectedly, a further decrease in seeding density so that cells were isolated from neighboring cells decreased proliferation.

Abstract: Experiments using microengineered substrates showed that E-cadherin engagement stimulated the peak in proliferation at intermediate seeding densities, and that the proliferation arrest at high densities did not involve E-cadherin, but rather resulted from a crowding-dependent decrease in cell spreading against the underlying substrate.

Abstract: Rac1 activity, which was induced by E-cadherin engagement specifically at intermediate seeding densities, was required for the cadherin-stimulated proliferation, and the control of Rac1 activation by E-cadherin was mediated by p120-catenin.

Abstract: Together, these findings demonstrate a stimulatory role for E-cadherin in proliferative regulation, and identify a simple mechanism by which cell–cell contact may trigger or inhibit epithelial cell proliferation in different settings.


Structure of Actin-related protein 8 and its contribution to nucleosome binding

Abstract: Nuclear actin-related proteins (Arps) are subunits of several chromatin remodelers, but their molecular functions within these complexes are unclear.

Abstract: We report the crystal structure of the INO80 complex subunit Arp8 in its ATP-bound form.

Abstract: Human Arp8 has several insertions in the conserved actin fold that explain its inability to polymerize.

Abstract: Most remarkably, one insertion wraps over the active site cleft and appears to rigidify the domain architecture, while active site features shared with actin suggest an allosterically controlled ATPase activity.

Abstract: Quantitative binding studies with nucleosomes and histone complexes reveal that Arp8 and the Arp8-Arp4-actin-HSA sub-complex of INO80 strongly prefer nucleosomes and H3-H4 tetramers over H2A-H2B dimers, suggesting that Arp8 functions as a nucleosome recognition module.

Abstract: In contrast, Arp4 prefers free (H3-H4)(2) over nucleosomes and may serve remodelers through binding to (dis)assembly intermediates in the remodeling reaction.


Regulation of β-Adrenergic Receptor Signaling by S-Nitrosylation of G-Protein-Coupled Receptor Kinase 2

Abstract: beta-adrenergic receptors (beta-ARs), prototypic G-protein-coupled receptors (GPCRs), play a critical role in regulating numerous physiological processes.

Abstract: The GPCR kinases (GRKs) curtail G-protein signaling and target receptors for internalization.

Abstract: Nitric oxide (NO) and/or S-nitrosothiols (SNOs) can prevent the loss of beta-AR signaling in vivo, but the molecular details are unknown.

Abstract: Here we show in mice that SNOs increase beta-AR expression and prevent agonist-stimulated receptor downregulation; and in cells, SNOs decrease GRK2-mediated beta-AR phosphorylation and subsequent recruitment of beta-arrestin to the receptor, resulting in the attenuation of receptor desensitization and internalization.

Abstract: In both cells and tissues, GRK2 is S-nitrosylated by SNOs as well as by NO synthases, and GRK2 S-nitrosylation increases following stimulation of multiple GPCRs with agonists.

Abstract: Cys340 of GRK2 is identified as a principal locus of inhibition by S-nitrosylation.

Abstract: Our studies thus reveal a central molecular mechanism through which GPCR signaling is regulated.


Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing

Abstract: Recent use of the CRISPR/Cas9 system has dramatically reduced the time required to produce mutant mice, but the involvement of a time-consuming microinjection step still hampers its application for high-throughput genetic analysis.

Abstract: Here we developed a simple, highly efficient, and large-scale genome editing method, in which the RNAs for the CRISPR/Cas9 system are electroporated into zygotes rather than microinjected.

Abstract: We used this method to perform single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in in mouse embryos.

Abstract: This method facilitates large-scale genetic analysis in the mouse.


Summary

Abstract: The genetic diseases Hutchinson-Gilford progeria syndrome (HGPS) and restrictive dermopathy (RD) arise from accumulation of farnesylated prelamin A because of defects in the lamin A maturation pathway.

Abstract: Both of these diseases exhibit symptoms that can be viewed as accelerated aging.

Abstract: The mechanism by which accumulation of farnesylated prelamin A leads to these accelerated aging phenotypes is not understood.

Abstract: Here we present evidence that in HGPS and RD fibroblasts, DNA damage checkpoints are persistently activated because of the compromise in genomic integrity.

Abstract: Inactivation of checkpoint kinases Ataxia-telangiectasia-mutated (ATM) and ATR (ATM- and Rad3-related) in these patient cells can partially overcome their early replication arrest.

Abstract: Treatment of patient cells with a protein farnesyltransferase inhibitor (FTI) did not result in reduction of DNA double-strand breaks and damage checkpoint signaling, although the treatment significantly reversed the aberrant shape of their nuclei.

Abstract: This suggests that DNA damage accumulation and aberrant nuclear morphology are independent phenotypes arising from prelamin A accumulation in these progeroid syndromes.

Abstract: Since DNA damage accumulation is an important contributor to the symptoms of HGPS, our results call into question the possibility of treatment of HGPS with FTIs alone.


The Buccaneer software for automated model building. 1. Tracing protein chains.

Abstract: A new technique for the automated tracing of protein chains in experimental electron-density maps is described.

Abstract: The technique relies on the repeated application of an oriented electron-density likelihood target function to identify likely C(alpha) positions.

Abstract: This function is applied both in the location of a few promising ;seed' positions in the map and to grow those initial C(alpha) positions into extended chain fragments.

Abstract: Techniques for assembling the chain fragments into an initial chain trace are discussed.


Expression profile of genes modulated by Aloe emodin in human U87 glioblastoma cells.

Abstract: Glioblastoma, the most aggressive and malignant form of glioma, appears to be resistant to various chemotherapeutic agents.

Abstract: Hence, approaches have been intensively investigated to targeti specific molecular pathways involved in glioblastoma development and progression.

Abstract: Aloe emodin is believed to modulate the expression of several genes in cancer cells.

Abstract: We aimed to understand the molecular mechanisms underlying the therapeutic effect of Aloe emodin on gene expression profiles in the human U87 glioblastoma cell line utilizing microarray technology.

Abstract: The gene expression analysis revealed that a total of 8,226 gene alterations out of 28,869 genes were detected after treatment with 58.6 μg/ml for 24 hours.

Abstract: Out of this total, 34 genes demonstrated statistically significant change (p<0.05) ranging from 1.07 to 1.87 fold.

Abstract: The results revealed that 22 genes were up-regulated and 12 genes were down-regulated in response to Aloe emodin treatment.

Abstract: These genes were then grouped into several clusters based on their biological functions, revealing induction of expression of genes involved in apoptosis (programmed cell death) and tissue remodelling in U87 cells (p<0.01).

Abstract: Several genes with significant changes of the expression level e.g. SHARPIN, BCAP31, FIS1, RAC1 and TGM2 from the apoptotic cluster were confirmed by quantitative real-time PCR (qRT-PCR).

Abstract: These results could serve as guidance for further studies in order to discover molecular targets for the cancer therapy based on Aloe emodin treatment.


The miniature genome of a carnivorous plant Genlisea aurea contains a low number of genes and short non-coding sequences

Abstract: BACKGROUND Genlisea aurea (Lentibulariaceae) is a carnivorous plant with unusually small genome size - 63.6 Mb - one of the smallest known among higher plants.

Abstract: Data on the genome sizes and the phylogeny of Genlisea suggest that this is a derived state within the genus.

Abstract: Thus, G. aurea is an excellent model organism for studying evolutionary mechanisms of genome contraction.

Abstract: RESULTS Here we report sequencing and de novo draft assembly of G. aurea genome.

Abstract: The assembly consists of 10,687 contigs of the total length of 43.4 Mb and includes 17,755 complete and partial protein-coding genes.

Abstract: Its comparison with the genome of Mimulus guttatus, another representative of higher core Lamiales clade, reveals striking differences in gene content and length of non-coding regions.

Abstract: CONCLUSIONS Genome contraction was a complex process, which involved gene loss and reduction of lengths of introns and intergenic regions, but not intron loss.

Abstract: The gene loss is more frequent for the genes that belong to multigenic families indicating that genetic redundancy is an important prerequisite for genome size reduction.


Abnormal Spine Morphology and Enhanced LTP in LIMK-1 Knockout Mice

Abstract: In vitro studies indicate a role for the LIM kinase family in the regulation of cofilin phosphorylation and actin dynamics.

Abstract: In addition, abnormal expression of LIMK-1 is associated with Williams syndrome, a mental disorder with profound deficits in visuospatial cognition.

Abstract: However, the in vivo function of this family of kinases remains elusive.

Abstract: Using LIMK-1 knockout mice, we demonstrate a significant role for LIMK-1 in vivo in regulating cofilin and the actin cytoskeleton.

Abstract: Furthermore, we show that the knockout mice exhibited significant abnormalities in spine morphology and in synaptic function, including enhanced hippocampal long-term potentiation.

Abstract: The knockout mice also showed altered fear responses and spatial learning.

Abstract: These results indicate that LIMK-1 plays a critical role in dendritic spine morphogenesis and brain function.


Wdr5 Mediates Self-Renewal and Reprogramming via the Embryonic Stem Cell Core Transcriptional Network

Abstract: The embryonic stem (ES) cell transcriptional and chromatin-modifying networks are critical for self-renewal maintenance.

Abstract: However, it remains unclear whether these networks functionally interact and, if so, what factors mediate such interactions.

Abstract: Here, we show that WD repeat domain 5 (Wdr5), a core member of the mammalian Trithorax (trxG) complex, positively correlates with the undifferentiated state and is a regulator of ES cell self-renewal.

Abstract: We demonstrate that Wdr5, an "effector" of H3K4 methylation, interacts with the pluripotency transcription factor Oct4.

Abstract: Genome-wide protein localization and transcriptome analyses demonstrate overlapping gene regulatory functions between Oct4 and Wdr5.

Abstract: The Oct4-Sox2-Nanog circuitry and trxG cooperate in activating transcription of key self-renewal regulators, and furthermore, Wdr5 expression is required for the efficient formation of induced pluripotent stem (iPS) cells.

Abstract: We propose an integrated model of transcriptional and epigenetic control, mediated by select trxG members, for the maintenance of ES cell self-renewal and somatic cell reprogramming.


Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson's disease.

Abstract: The long-term goal of nuclear transfer or alternative reprogramming approaches is to create patient-specific donor cells for transplantation therapy, avoiding immunorejection, a major complication in current transplantation medicine.

Abstract: It was recently shown that the four transcription factors Oct4, Sox2, Klf4, and c-Myc induce pluripotency in mouse fibroblasts.

Abstract: However, the therapeutic potential of induced pluripotent stem (iPS) cells for neural cell replacement strategies remained unexplored.

Abstract: Here, we show that iPS cells can be efficiently differentiated into neural precursor cells, giving rise to neuronal and glial cell types in culture.

Abstract: Upon transplantation into the fetal mouse brain, the cells migrate into various brain regions and differentiate into glia and neurons, including glutamatergic, GABAergic, and catecholaminergic subtypes.

Abstract: Electrophysiological recordings and morphological analysis demonstrated that the grafted neurons had mature neuronal activity and were functionally integrated in the host brain.

Abstract: Furthermore, iPS cells were induced to differentiate into dopamine neurons of midbrain character and were able to improve behavior in a rat model of Parkinson's disease upon transplantation into the adult brain.

Abstract: We minimized the risk of tumor formation from the grafted cells by separating contaminating pluripotent cells and committed neural cells using fluorescence-activated cell sorting.

Abstract: Our results demonstrate the therapeutic potential of directly reprogrammed fibroblasts for neuronal cell replacement in the animal model.


Responses of primate frontal cortex neurons during natural vocal communication.

Abstract: The role of primate frontal cortex in vocal communication and its significance in language evolution have a controversial history.

Abstract: While evidence indicates that vocalization processing occurs in ventrolateral prefrontal cortex neurons, vocal-motor activity has been conjectured to be primarily subcortical and suggestive of a distinctly different neural architecture from humans.

Abstract: Direct evidence of neural activity during natural vocal communication is limited, as previous studies were performed in chair-restrained animals.

Abstract: Here we recorded the activity of single neurons across multiple regions of prefrontal and premotor cortex while freely moving marmosets engaged in a natural vocal behavior known as antiphonal calling.

Abstract: Our aim was to test whether neurons in marmoset frontal cortex exhibited responses during vocal-signal processing and/or vocal-motor production in the context of active, natural communication.

Abstract: We observed motor-related changes in single neuron activity during vocal production, but relatively weak sensory responses for vocalization processing during this natural behavior.

Abstract: Vocal-motor responses occurred both prior to and during call production and were typically coupled to the timing of each vocalization pulse.

Abstract: Despite the relatively weak sensory responses a population classifier was able to distinguish between neural activity that occurred during presentations of vocalization stimuli that elicited an antiphonal response and those that did not.

Abstract: These findings are suggestive of the role that nonhuman primate frontal cortex neurons play in natural communication and provide an important foundation for more explicit tests of the functional contributions of these neocortical areas during vocal behaviors.


Salt-inducible kinase 2 links transcriptional coactivator p300 phosphorylation to the prevention of ChREBP-dependent hepatic steatosis in mice.

Abstract: Obesity and type 2 diabetes are associated with increased lipogenesis in the liver.

Abstract: This results in fat accumulation in hepatocytes, a condition known as hepatic steatosis, which is a form of nonalcoholic fatty liver disease (NAFLD), the most common cause of liver dysfunction in the United States.

Abstract: Carbohydrate-responsive element-binding protein (ChREBP), a transcriptional activator of glycolytic and lipogenic genes, has emerged as a major player in the development of hepatic steatosis in mice.

Abstract: However, the molecular mechanisms enhancing its transcriptional activity remain largely unknown.

Abstract: In this study, we have identified the histone acetyltransferase (HAT) coactivator p300 and serine/threonine kinase salt-inducible kinase 2 (SIK2) as key upstream regulators of ChREBP activity.

Abstract: In cultured mouse hepatocytes, we showed that glucose-activated p300 acetylated ChREBP on Lys672 and increased its transcriptional activity by enhancing its recruitment to its target gene promoters.

Abstract: SIK2 inhibited p300 HAT activity by direct phosphorylation on Ser89, which in turn decreased ChREBP-mediated lipogenesis in hepatocytes and mice overexpressing SIK2.

Abstract: Moreover, both liver-specific SIK2 knockdown and p300 overexpression resulted in hepatic steatosis, insulin resistance, and inflammation, phenotypes reversed by SIK2/p300 co-overexpression.

Abstract: Finally, in mouse models of type 2 diabetes and obesity, low SIK2 activity was associated with increased p300 HAT activity, ChREBP hyperacetylation, and hepatic steatosis.

Abstract: Our findings suggest that inhibition of hepatic p300 activity may be beneficial for treating hepatic steatosis in obesity and type 2 diabetes and identify SIK2 activators and specific p300 inhibitors as potential targets for pharmaceutical intervention.


A Role for NuSAP in Linking Microtubules to Mitotic Chromosomes

Abstract: The spindle apparatus is a microtubule (MT)-based machinery that attaches to and segregates the chromosomes during mitosis and meiosis.

Abstract: Self-organization of the spindle around chromatin involves the assembly of MTs, their attachment to the chromosomes, and their organization into a bipolar array.

Abstract: One regulator of spindle self-organization is RanGTP.

Abstract: RanGTP is generated at chromatin and activates a set of soluble, Ran-regulated spindle factors such as TPX2, NuMA, and NuSAP .

Abstract: How the spindle factors direct and attach MTs to the chromosomes are key open questions.

Abstract: Nucleolar and Spindle-Associated Protein (NuSAP) was recently identified as an essential MT-stabilizing and bundling protein that is enriched at the central part of the spindle .

Abstract: Here, we show by biochemical reconstitution that NuSAP efficiently adsorbs to isolated chromatin and DNA and that it can directly produce and retain high concentrations of MTs in the immediate vicinity of chromatin or DNA.

Abstract: Moreover, our data reveal that NuSAP-chromatin interaction is subject to Ran regulation and can be suppressed by Importin alpha (Impalpha) and Imp7.

Abstract: We propose that the presence of MT binding agents such as NuSAP, which can be directly immobilized on chromatin, are critical for targeting MT production to vertebrate chromosomes during spindle self-organization.


Molecular features of cellular reprogramming and development

Abstract: Differentiating somatic cells are progressively restricted to specialized functions during ontogeny, but they can be experimentally directed to form other cell types, including those with complete embryonic potential.

Abstract: Early nuclear reprogramming methods, such as somatic cell nuclear transfer (SCNT) and cell fusion, posed significant technical hurdles to precise dissection of the regulatory programmes governing cell identity.

Abstract: However, the discovery of reprogramming by ectopic expression of a defined set of transcription factors, known as direct reprogramming, provided a tractable platform to uncover molecular characteristics of cellular specification and differentiation, cell type stability and pluripotency.

Abstract: We discuss the control and maintenance of cellular identity during developmental transitions as they have been studied using direct reprogramming, with an emphasis on transcriptional and epigenetic regulation.


Small molecule modifiers of circadian clocks

Abstract: Circadian clocks orchestrate 24-h oscillations of essential physiological and behavioral processes in response to daily environmental changes.

Abstract: These clocks are remarkably precise under constant conditions yet highly responsive to resetting signals.

Abstract: With the molecular composition of the core oscillator largely established, recent research has increasingly focused on clock-modifying mechanisms/molecules.

Abstract: In particular, small molecule modifiers, intrinsic or extrinsic, are emerging as powerful tools for understanding basic clock biology as well as developing putative therapeutic agents for clock-associated diseases.

Abstract: In this review, we will focus on synthetic compounds capable of modifying the period, phase, or amplitude of circadian clocks, with particular emphasis on the mammalian clock.

Abstract: We will discuss the potential of exploiting these small molecule modifiers in both basic and translational research.


NVP-BEZ235, a dual PI3K/mTOR inhibitor, prevents PI3K signaling and inhibits the growth of cancer cells with activating PI3K mutations.

Abstract: Phosphatidylinositol-3-kinase (PI3K) pathway deregulation is a common event in human cancer, either through inactivation of the tumor suppressor phosphatase and tensin homologue deleted from chromosome 10 or activating mutations of p110-alpha.

Abstract: These hotspot mutations result in oncogenic activity of the enzyme and contribute to therapeutic resistance to the anti-HER2 antibody trastuzumab.

Abstract: The PI3K pathway is, therefore, an attractive target for cancer therapy.

Abstract: We have studied NVP-BEZ235, a dual inhibitor of the PI3K and the downstream mammalian target of rapamycin (mTOR).

Abstract: NVP-BEZ235 inhibited the activation of the downstream effectors Akt, S6 ribosomal protein, and 4EBP1 in breast cancer cells.

Abstract: The antiproliferative activity of NVP-BEZ235 was superior to the allosteric selective mTOR complex inhibitor everolimus in a panel of 21 cancer cell lines of different origin and mutation status.

Abstract: The described Akt activation due to mTOR inhibition was prevented by higher doses of NVP-BEZ235.

Abstract: NVP-BEZ235 reversed the hyperactivation of the PI3K/mTOR pathway caused by the oncogenic mutations of p110-alpha, E545K, and H1047R, and inhibited the proliferation of HER2-amplified BT474 cells exogenously expressing these mutations that render them resistant to trastuzumab.

Abstract: In trastuzumab-resistant BT474 H1047R breast cancer xenografts, NVP-BEZ235 inhibited PI3K signaling and had potent antitumor activity.

Abstract: In treated animals, there was complete inhibition of PI3K signaling in the skin at pharmacologically active doses, suggesting that skin may serve as surrogate tissue for pharmacodynamic studies.

Abstract: In summary, NVP-BEZ235 inhibits the PI3K/mTOR axis and results in antiproliferative and antitumoral activity in cancer cells with both wild-type and mutated p110-alpha.


Pharmacokinetic, pharmacodynamic, and pharmacogenetic determinants of osteonecrosis in children with acute lymphoblastic leukemia.

Abstract: Osteonecrosis is a severe glucocorticoid-induced complication of acute lymphoblastic leukemia treatment.

Abstract: We prospectively screened children (n = 364) with magnetic resonance imaging of hips and knees, regardless of symptoms; the cumulative incidence of any (grade 1-4) versus symptomatic (grade 2-4) osteonecrosis was 71.8% versus 17.6%, respectively.

Abstract: We investigated whether age, race, sex, acute lymphoblastic leukemia treatment arm, body mass, serum lipids, albumin and cortisol levels, dexamethasone pharmacokinetics, and genome-wide germline genetic polymorphisms were associated with symptomatic osteonecrosis.

Abstract: Age more than 10 years (odds ratio, = 4.85; 95% confidence interval, 2.5-9.2; P = .00001) and more intensive treatment (odds ratio = 2.5; 95% confidence interval, 1.2-4.9; P = .011) were risk factors and included as covariates in all analyses.

Abstract: Lower albumin (P = .05) and elevated cholesterol (P = .02) associated with symptomatic osteonecrosis, and severe (grade 3 or 4) osteonecrosis was linked to poor dexamethasone clearance (P = .0005).

Abstract: Adjusting for clinical features, polymorphisms of ACP1 (eg, rs12714403, P = 1.9 × 10(-6), odds ratio = 5.6; 95% confidence interval, 2.7-11.3), which regulates lipid levels and osteoblast differentiation, were associated with risk of osteonecrosis as well as with lower albumin and higher cholesterol.

Abstract: Overall, older age, lower albumin, higher lipid levels, and dexamethasone exposure were associated with osteonecrosis and may be linked by inherited genomic variation.


Prevention of Rectal SHIV Transmission in Macaques by Daily or Intermittent Prophylaxis with Emtricitabine and Tenofovir

Abstract: BACKGROUND In the absence of an effective vaccine, HIV continues to spread globally, emphasizing the need for novel strategies to limit its transmission.

Abstract: Pre-exposure prophylaxis (PrEP) with antiretroviral drugs could prove to be an effective intervention strategy if highly efficacious and cost-effective PrEP modalities are identified.

Abstract: We evaluated daily and intermittent PrEP regimens of increasing antiviral activity in a macaque model that closely resembles human transmission.

Abstract: METHODS AND FINDINGS We used a repeat-exposure macaque model with 14 weekly rectal virus challenges.

Abstract: Three drug treatments were given once daily, each to a different group of six rhesus macaques.

Abstract: Group 1 was treated subcutaneously with a human-equivalent dose of emtricitabine (FTC), group 2 received orally the human-equivalent dosing of both FTC and tenofovir-disoproxil fumarate (TDF), and group 3 received subcutaneously a similar dosing of FTC and a higher dose of tenofovir.

Abstract: A fourth group of six rhesus macaques (group 4) received intermittently a PrEP regimen similar to group 3 only 2 h before and 24 h after each weekly virus challenge.

Abstract: Results were compared to 18 control macaques that did not receive any drug treatment.

Abstract: The risk of infection in macaques treated in groups 1 and 2 was 3.8- and 7.8-fold lower than in untreated macaques (p = 0.02 and p = 0.008, respectively).

Abstract: All six macaques in group 3 were protected.

Abstract: Breakthrough infections had blunted acute viremias; drug resistance was seen in two of six animals.

Abstract: All six animals in group 4 that received intermittent PrEP were protected.

Abstract: CONCLUSIONS This model suggests that single drugs for daily PrEP can be protective but a combination of antiretroviral drugs may be required to increase the level of protection.

Abstract: Short but potent intermittent PrEP can provide protection comparable to that of daily PrEP in this SHIV/macaque model.

Abstract: These findings support PrEP trials for HIV prevention in humans and identify promising PrEP modalities.


Direct-coupling analysis of residue co-evolution captures native contacts across many protein families

Abstract: The similarity in the three-dimensional structures of homologous proteins imposes strong constraints on their sequence variability.

Abstract: It has long been suggested that the resulting correlations among amino acid compositions at different sequence positions can be exploited to infer spatial contacts within the tertiary protein structure.

Abstract: Crucial to this inference is the ability to disentangle direct and indirect correlations, as accomplished by the recently introduced Direct Coupling Analysis (DCA) (Weigt et al. (2009) Proc Natl Acad Sci 106:67).

Abstract: Here we develop a computationally efficient implementation of DCA, which allows us to evaluate the accuracy of contact prediction by DCA for a large number of protein domains, based purely on sequence information.

Abstract: DCA is shown to yield a large number of correctly predicted contacts, recapitulating the global structure of the contact map for the majority of the protein domains examined.

Abstract: Furthermore, our analysis captures clear signals beyond intra- domain residue contacts, arising, e.g., from alternative protein conformations, ligand- mediated residue couplings, and inter-domain interactions in protein oligomers.

Abstract: Our findings suggest that contacts predicted by DCA can be used as a reliable guide to facilitate computational predictions of alternative protein conformations, protein complex formation, and even the de novo prediction of protein domain structures, provided the existence of a large number of homologous sequences which are being rapidly made available due to advances in genome sequencing.


Variation in antibiotic prescribing and its impact on recovery in patients with acute cough in primary care: prospective study in 13 countries.

Abstract: OBJECTIVE To describe variation in antibiotic prescribing for acute cough in contrasting European settings and the impact on recovery.

Abstract: DESIGN Cross sectional observational study with clinicians from 14 primary care research networks in 13 European countries who recorded symptoms on presentation and management.

Abstract: Patients followed up for 28 days with patient diaries.

Abstract: SETTING Primary care.

Abstract: PARTICIPANTS Adults with a new or worsening cough or clinical presentation suggestive of lower respiratory tract infection.

Abstract: MAIN OUTCOME MEASURES Prescribing of antibiotics by clinicians and total symptom severity scores over time.

Abstract: RESULTS 3402 patients were recruited (clinicians completed a case report form for 99% (3368) of participants and 80% (2714) returned a symptom diary).

Abstract: Mean symptom severity scores at presentation ranged from 19 (scale range 0 to 100) in networks based in Spain and Italy to 38 in the network based in Sweden.

Abstract: Antibiotic prescribing by networks ranged from 20% to nearly 90% (53% overall), with wide variation in classes of antibiotics prescribed.

Abstract: Amoxicillin was overall the most common antibiotic prescribed, but this ranged from 3% of antibiotics prescribed in the Norwegian network to 83% in the English network.

Abstract: While fluoroquinolones were not prescribed at all in three networks, they were prescribed for 18% in the Milan network.

Abstract: After adjustment for clinical presentation and demographics, considerable differences remained in antibiotic prescribing, ranging from Norway (odds ratio 0.18, 95% confidence interval 0.11 to 0.30) to Slovakia (11.2, 6.20 to 20.27) compared with the overall mean (proportion prescribed: 0.53).

Abstract: The rate of recovery was similar for patients who were and were not prescribed antibiotics (coefficient -0.01, P<0.01) once clinical presentation was taken into account.

Abstract: CONCLUSIONS Variation in clinical presentation does not explain the considerable variation in antibiotic prescribing for acute cough in Europe.

Abstract: Variation in antibiotic prescribing is not associated with clinically important differences in recovery.

Abstract: TRIAL REGISTRATION Clinicaltrials.gov NCT00353951.


Dorsomorphin, a Selective Small Molecule Inhibitor of BMP Signaling, Promotes Cardiomyogenesis in Embryonic Stem Cells

Abstract: BACKGROUND Pluripotent embryonic stem (ES) cells, which have the capacity to give rise to all tissue types in the body, show great promise as a versatile source of cells for regenerative therapy.

Abstract: However, the basic mechanisms of lineage specification of pluripotent stem cells are largely unknown, and generating sufficient quantities of desired cell types remains a formidable challenge.

Abstract: Small molecules, particularly those that modulate key developmental pathways like the bone morphogenetic protein (BMP) signaling cascade, hold promise as tools to study in vitro lineage specification and to direct differentiation of stem cells toward particular cell types.

Abstract: METHODOLOGY/ PRINCIPAL FINDINGS We describe the use of dorsomorphin, a selective small molecule inhibitor of BMP signaling, to induce myocardial differentiation in mouse ES cells.

Abstract: Cardiac induction is very robust, increasing the yield of spontaneously beating cardiomyocytes by at least 20 fold.

Abstract: Dorsomorphin, unlike the endogenous BMP antagonist Noggin, robustly induces cardiomyogenesis when treatment is limited to the initial 24-hours of ES cell differentiation.

Abstract: Quantitative-PCR analyses of differentiating ES cells indicate that pharmacological inhibition of BMP signaling during the early critical stage promotes the development of the cardiomyocyte lineage, but reduces the differentiation of endothelial, smooth muscle, and hematopoietic cells.

Abstract: CONCLUSIONS/ SIGNIFICANCE Administration of a selective small molecule BMP inhibitor during the initial stages of ES cell differentiation substantially promotes the differentiation of primitive pluripotent cells toward the cardiomyocytic lineage, apparently at the expense of other mesodermal lineages.

Abstract: Small molecule modulators of developmental pathways like dorsomorphin could become versatile pharmacological tools for stem cell research and regenerative medicine.


Cineradiography of Monkey Lip-Smacking Reveals Putative Precursors of Speech Dynamics

Abstract: A key feature of speech is its stereotypical 5 Hz rhythm.

Abstract: One theory posits that this rhythm evolved through the modification of rhythmic facial movements in ancestral primates.

Abstract: If the hypothesis has any validity, then a comparative approach may shed some light.

Abstract: We tested this idea by using cineradiography (X-ray movies) to characterize and quantify the internal dynamics of the macaque monkey vocal tract during lip-smacking (a rhythmic facial expression) versus chewing.

Abstract: Previous human studies showed that speech movements are faster than chewing movements, and the functional coordination between vocal tract structures is different between the two behaviors.

Abstract: If rhythmic speech evolved through a rhythmic ancestral facial movement, then one hypothesis is that monkey lip-smacking versus chewing should also exhibit these differences.

Abstract: We found that the lips, tongue, and hyoid move with a speech-like 5 Hz rhythm during lip-smacking, but not during chewing.

Abstract: Most importantly, the functional coordination between these structures was distinct for each behavior.

Abstract: These data provide empirical support for the idea that the human speech rhythm evolved from the rhythmic facial expressions of ancestral primates.


Interneuronal DISC1 regulates NRG1-ErbB4 signalling and excitatory-inhibitory synapse formation in the mature cortex.

Abstract: Neuregulin-1 (NRG1) and its receptor ErbB4 influence several processes of neurodevelopment, but the mechanisms regulating this signalling in the mature brain are not well known.

Abstract: DISC1 is a multifunctional scaffold protein that mediates many cellular processes.

Abstract: Here we present a functional relationship between DISC1 and NRG1-ErbB4 signalling in mature cortical interneurons.

Abstract: By cell type-specific gene modulation in vitro and in vivo including in a mutant DISC1 mouse model, we demonstrate that DISC1 inhibits NRG1-induced ErbB4 activation and signalling.

Abstract: This effect is likely mediated by competitive inhibition of binding of ErbB4 to PSD95.

Abstract: Finally, we show that interneuronal DISC1 affects NRG1-ErbB4-mediated phenotypes in the fast spiking interneuron-pyramidal neuron circuit.

Abstract: Post-mortem brain analyses and some genetic studies have reported interneuronal deficits and involvement of the DISC1, NRG1 and ErbB4 genes in schizophrenia, respectively.

Abstract: Our results suggest a mechanism by which cross-talk between DISC1 and NRG1-ErbB4 signalling may contribute to these deficits.


The Proto-Oncogene c-maf Is Responsible for Tissue-Specific Expression of Interleukin-4

Abstract: The molecular basis for the distinctive cytokine expression of CD4+ T helper 1 (Th1) and T helper 2 (Th2) subsets remains elusive.

Abstract: Here, we report that the proto-oncogene c-maf, a basic region/leucine zipper transcription factor, controls tissue-specific expression of IL-4.

Abstract: c-Maf is expressed in Th2 but not Th1 clones and is induced during normal precursor cell differentiation along a Th2 but not Th1 lineage.

Abstract: c-Maf binds to a c-Maf response element (MARE) in the proximal IL-4 promoter adjacent to a site footprinted by extracts from Th2 but not Th1 clones.

Abstract: Ectopic expression of c-Maf transactivates the IL-4 promoter in Th1 cells, B cells, and nonlymphoid cells, a function that maps to the MARE and Th2-specific footprint.

Abstract: Furthermore, c-Maf acts in synergy with the nuclear factor of activated T cells (NF-ATp) to initiate endogeneous IL-4 production by B cells.

Abstract: Manipulation of c-Maf may alter Th subset ratios in human disease.


hClock gene expression in human colorectal carcinoma.

Abstract: In this study, we aimed to investigate changes in the expression of human Clock (hClock), a gene at the core of the circadian gene family, in colorectal carcinomas (CRCs) and to discuss the possible effects.

Abstract: Previous studies have revealed that the disruption of circadian rhythms is one of the endogenous factors that contribute to the initiation and development of CRCs.

Abstract: However, the underlying molecular changes to the circadian genes associated with CRCs have not been explored.

Abstract: Immunofluorescence and quantitative polymerase chain reaction (qPCR) analysis of the hCLOCK protein and gene expression were performed in 30 cases of CRC.

Abstract: The hCLOCK protein was expressed in all specimens obtained from 30 CRC patients.

Abstract: Higher levels of hCLOCK expression were observed in human CRC tissues compared with the paired non-cancerous tissues.

Abstract: hCLOCK expression was significantly higher in poorly differentiated, or late-stage, Dukes' grade tumors and in 64.3% of tumor cases with lymph node metastasis.

Abstract: The hClock gene was expressed in all specimens.

Abstract: A significantly higher expression of hClock was found in human CRC cases compared with paired non-cancerous tissues.

Abstract: There was a strong positive linear correlation between hClock gene expression and protein expression in human CRCs.

Abstract: A strong positive linear correlation was also found between hClock gene expression and ARNT, HIF-1α and VEGF expression in human CRCs.

Abstract: There was no significant correlation between hClock and Bak, Bax, Bid, tumor necrosis factor receptor I (TNFR I) and TNFR II.

Abstract: The circadian gene hClock was stably expressed in human colorectal mucosa and was important in regulating the expression of downstream clock-controlled genes.

Abstract: hCLOCK may interact with HIF-1α/ARNT and activate VEGF to stimulate tumor angiogenesis and metastasis.


The Antiretroviral Therapy in Lower Income Countries (ART-LINC) Collaboration and ART Cohort Collaboration (ART-CC) groups Summary

Abstract: BACKGROUND Highly active antiretroviral therapy (HAART) is being scaled up in developing countries.

Abstract: We compared baseline characteristics and outcomes during the first year of HAART between HIV-1-infected patients in low-income and high-income settings.

Abstract: METHODS 18 HAART programmes in Africa, Asia, and South America (low-income settings) and 12 HIV cohort studies from Europe and North America (high-income settings) provided data for 4810 and 22,217, respectively, treatment-naïve adult patients starting HAART.

Abstract: All patients from high-income settings and 2725 (57%) patients from low-income settings were actively followed-up and included in survival analyses.

Abstract: FINDINGS Compared with high-income countries, patients starting HAART in low-income settings had lower CD4 cell counts (median 108 cells per muL vs 234 cells per muL), were more likely to be female (51%vs 25%), and more likely to start treatment with a non-nucleoside reverse transcriptase inhibitor (NNRTI) (70%vs 23%).

Abstract: At 6 months, the median number of CD4 cells gained (106 cells per muL vs 103 cells per muL) and the percentage of patients reaching HIV-1 RNA levels lower than 500 copies/mL (76%vs 77%) were similar.

Abstract: Mortality was higher in low-income settings (124 deaths during 2236 person-years of follow-up) than in high-income settings (414 deaths during 20,532 person-years).

Abstract: The adjusted hazard ratio (HR) of mortality comparing low-income with high-income settings fell from 4.3 (95% CI 1.6-11.8) during the first month to 1.5 (0.7-3.0) during months 7-12.

Abstract: The provision of treatment free of charge in low-income settings was associated with lower mortality (adjusted HR 0.23; 95% CI 0.08-0.61).

Abstract: INTERPRETATION Patients starting HAART in resource-poor settings have increased mortality rates in the first months on therapy, compared with those in developed countries.

Abstract: Timely diagnosis and assessment of treatment eligibility, coupled with free provision of HAART, might reduce this excess mortality.


A family of phosphoinositide 3-kinases in Drosophila identifies a new mediator of signal transduction

Abstract: BACKGROUND Mammalian phosphoinositide 3-kinases (PI 3-kinases) are involved in receptor-mediated signal transduction and have been implicated in processes such as transformation and mitogenesis through their role in elevating cellular phosphatidylinositol (3,4,5)-trisphosphate.

Abstract: Additionally, a PI 3-kinase activity which generates phosphatidylinositol 3-phosphate has been shown to be required for protein trafficking in yeast.

Abstract: RESULTS We have identified a family of three distinct PI 3-kinases in Drosophila, using an approach based on the polymerase chain reaction to amplify a region corresponding to the conserved catalytic domain of PI 3-kinases.

Abstract: One of these family members, PI3K_92D, is closely related to the prototypical PI 3-kinase, p110 alpha; PI3K_59F is homologous to Vps34p, whereas the third, PI3K_68D, is a novel PI 3-kinase which is widely expressed throughout the Drosophila life cycle.

Abstract: The PI3K_68D cDNA encodes a protein of 210 kDa, which lacks sequences implicated in linking p110 PI 3-kinases to p85 adaptor proteins, but contains an amino-terminal proline-rich sequence, which could bind to SH3 domains, and a carboxy-terminal C2 domain.

Abstract: Biochemical analyses demonstrate that PI3K_68D has a novel substrate specificity in vitro, restricted to phosphatidylinositol and phosphatidylinositol 4-phosphate, and is unable to phosphorylate phosphatidylinositol (4,5)-bisphosphate, the implied in vivo substrate for p110.

Abstract: CONCLUSIONS A family of PI 3-kinases in Drosophila, including a novel class represented by PI3K_68D, is described.

Abstract: PI3K_68D has the potential to bind to signalling molecules containing SH3 domains, lacks p85-adaptor-binding sequences, has a Ca(2+)-independent phospholipid-binding domain and displays a restricted in vitro substrate specificity, so it could define a novel signal transduction pathway.


Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks

Abstract: Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks.

Abstract: To better understand BIR, we performed a targeted siRNA screen for genes whose depletion inhibited G1 to S phase progression when oncogenic cyclin E was overexpressed.

Abstract: RAD52, a gene dispensable for normal development in mice, was among the top hits.

Abstract: In cells in which fork collapse was induced by oncogenes or chemicals, the Rad52 protein localized to DRS foci.

Abstract: Depletion of Rad52 by siRNA or knockout of the gene by CRISPR/Cas9 compromised restart of collapsed forks and led to DNA damage in cells experiencing DRS.

Abstract: Furthermore, in cancer-prone, heterozygous APC mutant mice, homozygous deletion of the Rad52 gene suppressed tumor growth and prolonged lifespan.

Abstract: We therefore propose that mammalian RAD52 facilitates repair of collapsed DNA replication forks in cancer cells.