Effect of low doses of ionising radiation in infancy on cognitive function in adulthood: Swedish population based cohort study.

Abstract: OBJECTIVE To determine whether exposure to low doses of ionising radiation in infancy affects cognitive function in adulthood.

Abstract: DESIGN Population based cohort study.

Abstract: SETTING Sweden.

Abstract: PARTICIPANTS 3094 men who had received radiation for cutaneous haemangioma before age 18 months during 1930-59.

Abstract: MAIN OUTCOME MEASURES Radiation dose to frontal and posterior parts of the brain, and association between dose and intellectual capacity at age 18 or 19 years based on cognitive tests (learning ability, logical reasoning, spatial recognition) and high school attendance.

Abstract: RESULTS The proportion of boys who attended high school decreased with increasing doses of radiation to both the frontal and the posterior parts of the brain from about 32% among those not exposed to around 17% in those who received > 250 mGy.

Abstract: For the frontal dose, the multivariate odds ratio was 0.47 (95% confidence interval 0.26 to 0.85, P for trend 0.0003) and for the posterior dose it was 0.59 (0.23 to 1.47, 0.0005).

Abstract: A negative dose-response relation was also evident for the three cognitive tests for learning ability and logical reasoning but not for the test of spatial recognition.

Abstract: CONCLUSIONS Low doses of ionising radiation to the brain in infancy influence cognitive abilities in adulthood.


Differential effects of charybdotoxin on the activity of retinal ganglion cells in the dark- and light-adapted mouse retina

Abstract: Patch-clamp recordings were made from retinal ganglion cells in the mouse retina.

Abstract: Under dark adaptation, blockage of BK(Ca) channels increases the spontaneous excitatory postsynaptic currents (EPSCs) and light-evoked On-EPSCs, while it decreases the light-evoked Off inhibitory postsynaptic currents (IPSCs).

Abstract: However, under light adaptation it decreases the light-evoked On-EPSCs, the spontaneous IPSCs and the light-evoked On- and Off-IPSCs.

Abstract: Blockage of BK(Ca) channels significantly altered the outputs of RGCs by changing their light-evoked responses into a bursting pattern and increasing the light-evoked depolarization of the membrane potentials, while it did not significantly change the peak firing rates of light-evoked responses.


Hutchinson-Gilford progeria syndrome: review of the phenotype

Abstract: Hutchinson-Gilford progeria syndrome (HGPS) is a rare but well known entity characterized by extreme short stature, low body weight, early loss of hair, lipodystrophy, scleroderma, decreased joint mobility, osteolysis, and facial features that resemble aged persons.

Abstract: Cardiovascular compromise leads to early demise.

Abstract: Cognitive development is normal.

Abstract: Data on 10 of our own cases and 132 cases from literature are presented.

Abstract: The incidence in the last century in the Netherlands was 1:4,000,000.

Abstract: Sex ratio was 1.2:1.

Abstract: Main first symptoms were failure to thrive (55%), hair loss (40%), skin problems (28%), and lipodystrophy (20%).

Abstract: Mean age at diagnosis was 2.9 years.

Abstract: Growth in weight was more disturbed than growth in height, and growth delay started already prenatally.

Abstract: Mean height > 13 years was 109.0 cm, mean weight was 14.5 kg.

Abstract: Osteolysis was wide-spread but not expressed, except in the viscerocranium, and remained limited to membranous formed bone.

Abstract: Lipodystrophy is generalized, only intra-abdominal fat depositions remain present.

Abstract: Cardiovascular problems are extremely variable, both in age of onset and nature.

Abstract: Stroke and coronary dysfunctioning are most frequent.

Abstract: Pathologic findings in coronaries and aorta resemble sometimes the findings in elderly persons, but can also be much more limited.

Abstract: Loss of smooth muscle cells seems the most important finding.

Abstract: Mean age of demise was 12.6 years.

Abstract: Patients can be subdivided in patients with classical HGPS, which follows an autosomal dominant pattern of inheritance, (almost) all cases representing spontaneous mutations, and in non-classical progeria, in whom growth can be less retarded, scalp hair remains present for a longer time, lipodystrophy is more slowly progressive, osteolysis is more expressed except in the face, and survival well into adulthood is not uncommon.

Abstract: Pattern of inheritance of non-classical progeria is most probably autosomal recessive.

Abstract: The cause of HGPS is an abnormally formed Lamin A, either directly by a mutated LMNA gene, or through abnormal posttranslational processing (ZMPSTE24 gene mutations).

Abstract: Of 34 LMNA mutations found in progeria patients, there were 26 classical p.

Abstract: G608G mutations (76%).

Abstract: Pathogenesis is most likely to follow several different pathways.

Abstract: Potential therapeutic strategies are developed along these lines and include RNA interference techniques and inhibition of the dominant-negative influence of abnormally formed Lamin A on polymerization with normally formed Lamin A.


Identification of an autoantigen demonstrates a link between interstitial lung disease and a defect in central tolerance.

Abstract: Interstitial lung disease (ILD) is a common manifestation of systemic autoimmunity characterized by progressive inflammation or scarring of the lungs.

Abstract: Patients who develop these complications can exhibit significantly impaired gas exchange that may result in hypoxemia, pulmonary hypertension, and even death.

Abstract: Unfortunately, little is understood about how these diseases arise, including the role of specific defects in immune tolerance.

Abstract: Another key question is whether autoimmune responses targeting the lung parenchyma are critical to ILD pathogenesis, including that of isolated idiopathic forms.

Abstract: We show that a specific defect in central tolerance brought about by mutations in the autoimmune regulator gene (Aire) leads to an autoreactive T cell response to a lung antigen named vomeromodulin and the development of ILD.

Abstract: We found that a human patient and mice with defects in Aire develop similar lung pathology, demonstrating that the AIRE-deficient model of autoimmunity is a suitable translational system in which to unravel fundamental mechanisms of ILD pathogenesis.


In vivo role of ER-associated peptidase activity in tailoring peptides for presentation by MHC class Ia and class Ib molecules

Abstract: Endoplasmic reticulum (ER)-associated aminopeptidase (ERAP)1 has been implicated in the final proteolytic processing of peptides presented by major histocompatibility complex (MHC) class I molecules.

Abstract: To evaluate the in vivo role of ERAP1, we have generated ERAP1-deficient mice.

Abstract: Cell surface expression of the class Ia molecules H-2Kb and H-2Db and of the class Ib molecule Qa-2 was significantly reduced in these animals.

Abstract: Although cells from mutant animals exhibited reduced capacity to present several self- and foreign antigens to Kb-, Db-, or Qa-1b–restricted CD8+ cytotoxic T cells, presentation of some antigens was unaffected or significantly enhanced.

Abstract: Consistent with these findings, mice generated defective CD8+ T cell responses against class I–presented antigens.

Abstract: These findings reveal an important in vivo role of ER-associated peptidase activity in tailoring peptides for presentation by MHC class Ia and class Ib molecules.


PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro

Abstract: INTRODUCTION Alterations in cell cycle regulators have been implicated in human malignancies including breast cancer.

Abstract: PD 0332991 is an orally active, highly selective inhibitor of the cyclin D kinases (CDK)4 and CDK6 with ability to block retinoblastoma (Rb) phosphorylation in the low nanomolar range.

Abstract: To identify predictors of response, we determined the in vitro sensitivity to PD 0332991 across a panel of molecularly characterized human breast cancer cell lines.

Abstract: METHODS Forty-seven human breast cancer and immortalized cell lines representing the known molecular subgroups of breast cancer were treated with PD 0332991 to determine IC50 values.

Abstract: These data were analyzed against baseline gene expression data to identify genes associated with PD 0332991 response.

Abstract: RESULTS Cell lines representing luminal estrogen receptor-positive (ER+) subtype (including those that are HER2 amplified) were most sensitive to growth inhibition by PD 0332991 while nonluminal/basal subtypes were most resistant.

Abstract: Analysis of variance identified 450 differentially expressed genes between sensitive and resistant cells.

Abstract: pRb and cyclin D1 were elevated and CDKN2A (p16) was decreased in the most sensitive lines.

Abstract: Cell cycle analysis showed G0/G1 arrest in sensitive cell lines and Western blot analysis demonstrated that Rb phosphorylation is blocked in sensitive lines but not resistant lines.

Abstract: PD 0332991 was synergistic with tamoxifen and trastuzumab in ER+ and HER2-amplified cell lines, respectively.

Abstract: PD 0332991 enhanced sensitivity to tamoxifen in cell lines with conditioned resistance to ER blockade.

Abstract: CONCLUSIONS These studies suggest a role for CDK4/6 inhibition in some breast cancers and identify criteria for patient selection in clinical studies of PD 0332991


Identification of secreted proteins that mediate cell-cell interactions in an in vitro model of the lung cancer microenvironment.

Abstract: Non-small cell lung cancer (NSCLC) cells with somatic mutations in K-ras recruit to the tumor a variety of cell types (hereafter collectively termed "stromal cells") that can promote or inhibit tumorigenesis by mechanisms that have not been fully elucidated.

Abstract: Here, we postulated that stromal cells in the tumor microenvironment alter the tumor cell secretome, including those proteins required for tumor growth and dissemination, and we developed an in vitro model to test this hypothesis.

Abstract: Coculturing a murine K-ras mutant lung adenocarcinoma cell line (LKR-13) with a murine lung stromal cell (macrophage, endothelial cell, or fibroblast) enhanced stromal cell migration, induced endothelial tube formation, increased LKR-13 cell proliferation, and regulated the secretion of proteins involved in angiogenesis, inflammation, cell proliferation, and epithelial-to-mesenchymal transition.

Abstract: Among these proteins, CXCL1 has been reported to promote NSCLC development, whereas interleukin-18 (IL-18) has an undefined role.

Abstract: Genetic and pharmacologic strategies to inhibit CXCL1 and IL-18 revealed that stromal cell migration, LKR-13 cell proliferation, and LKR-13 cell tumorigenicity required one or both of these proteins.

Abstract: We conclude that stromal cells enhanced LKR-13 cell tumorigenicity partly through their effects on the secretome of LKR-13 cells.

Abstract: Strategies to inhibit tumor/stromal cell interactions may be useful as therapeutic approaches in NSCLC patients.


Tightrope act: autophagy in stem cell renewal, differentiation, proliferation, and aging

Abstract: Autophagy is a constitutive lysosomal catabolic pathway that degrades damaged organelles and protein aggregates.

Abstract: Stem cells are characterized by self-renewal, pluripotency, and quiescence; their long life span, limited capacity to dilute cellular waste and spent organelles due to quiescence, along with their requirement for remodeling in order to differentiate, all suggest that they require autophagy more than other cell types.

Abstract: Here, we review the current literature on the role of autophagy in embryonic and adult stem cells, including hematopoietic, mesenchymal, and neuronal stem cells, highlighting the diverse and contrasting roles autophagy plays in their biology.

Abstract: Furthermore, we review the few studies on stem cells, lysosomal activity, and autophagy.

Abstract: Novel techniques to detect autophagy in primary cells are required to study autophagy in different stem cell types.

Abstract: These will help to elucidate the importance of autophagy in stem cells during transplantation, a promising therapeutic approach for many diseases.


Variant Histone H2A.Z Is Globally Localized to the Promoters of Inactive Yeast Genes and Regulates Nucleosome Positioning

Abstract: H2A.Z is an evolutionary conserved histone variant involved in transcriptional regulation, antisilencing, silencing, and genome stability.

Abstract: The mechanism(s) by which H2A.Z regulates these various biological functions remains poorly defined, in part due to the lack of knowledge regarding its physical location along chromosomes and the bearing it has in regulating chromatin structure.

Abstract: Here we mapped H2A.Z across the yeast genome at an approximately 300-bp resolution, using chromatin immunoprecipitation combined with tiling microarrays.

Abstract: We have identified 4,862 small regions—typically one or two nucleosomes wide—decorated with H2A.Z. Those “Z loci” are predominantly found within specific nucleosomes in the promoter of inactive genes all across the genome.

Abstract: Furthermore, we have shown that H2A.Z can regulate nucleosome positioning at the GAL1 promoter.

Abstract: Within HZAD domains, the regions where H2A.Z shows an antisilencing function, H2A.Z is localized in a wider pattern, suggesting that the variant histone regulates a silencing and transcriptional activation via different mechanisms.

Abstract: Our data suggest that the incorporation of H2A.Z into specific promoter-bound nucleosomes configures chromatin structure to poise genes for transcriptional activation.

Abstract: The relevance of these findings to higher eukaryotes is discussed.


Collective trauma in northern Sri Lanka: a qualitative psychosocial-ecological study

Abstract: BACKGROUND Complex situations that follow war and natural disasters have a psychosocial impact on not only the individual but also on the family, community and society.

Abstract: Just as the mental health effects on the individual psyche can result in non pathological distress as well as a variety of psychiatric disorders; massive and widespread trauma and loss can impact on family and social processes causing changes at the family, community and societal levels.

Abstract: METHOD This qualitative, ecological study is a naturalistic, psychosocial ethnography in Northern Sri Lanka, while actively involved in psychosocial and community mental health programmes among the Tamil community.

Abstract: Participatory observation, key informant interviews and focus group discussion with community level relief and rehabilitation workers and government and non-governmental officials were used to gather data.

Abstract: The effects on the community of the chronic, man-made disaster, war, in Northern Sri Lanka were compared with the contexts found before the war and after the tsunami.

Abstract: RESULTS Fundamental changes in the functioning of the family and the community were observed.

Abstract: While the changes after the tsunami were not so prominent, the chronic war situation caused more fundamental social transformations.

Abstract: At the family level, the dynamics of single parent families, lack of trust among members, and changes in significant relationships, and child rearing practices were seen.

Abstract: Communities tended to be more dependent, passive, silent, without leadership, mistrustful, and suspicious.

Abstract: Additional adverse effects included the breakdown in traditional structures, institutions and familiar ways of life, and deterioration in social norms and ethics.

Abstract: A variety of community level interventions were tried.

Abstract: CONCLUSION Exposure to conflict, war and disaster situations impact on fundamental family and community dynamics resulting in changes at a collective level.

Abstract: Relief, rehabilitation and development programmes to be effective will need to address the problem of collective trauma, particularly using integrated multi-level approaches.


Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference.

Abstract: In the present study, the relationship between short interfering RNA (siRNA) sequence and RNA interference (RNAi) effect was extensively analyzed using 62 targets of four exogenous and two endogenous genes and three mammalian and Drosophila cells.

Abstract: We present the rules that may govern siRNA sequence preference and in accordance with which highly effective siRNAs essential for systematic mammalian functional genomics can be readily designed.

Abstract: These rules indicate that siRNAs which simultaneously satisfy all four of the following sequence conditions are capable of inducing highly effective gene silencing in mammalian cells: (i) A/U at the 5' end of the antisense strand; (ii) G/C at the 5' end of the sense strand; (iii) at least five A/U residues in the 5' terminal one-third of the antisense strand; and (iv) the absence of any GC stretch of more than 9 nt in length.

Abstract: siRNAs opposite in features with respect to the first three conditions give rise to little or no gene silencing in mammalian cells.

Abstract: Essentially the same rules for siRNA sequence preference were found applicable to DNA-based RNAi in mammalian cells and in ovo RNAi using chick embryos.

Abstract: In contrast to mammalian and chick cells, little siRNA sequence preference could be detected in Drosophila in vivo RNAi.


Overview of the CCP4 suite and current developments

Abstract: The CCP4 (Collaborative Computational Project, Number 4) software suite is a collection of programs and associated data and software libraries which can be used for macromolecular structure determination by X-ray crystallography.

Abstract: The suite is designed to be flexible, allowing users a number of methods of achieving their aims.

Abstract: The programs are from a wide variety of sources but are connected by a common infrastructure provided by standard file formats, data objects and graphical interfaces.

Abstract: Structure solution by macromolecular crystallography is becoming increasingly automated and the CCP4 suite includes several automation pipelines.

Abstract: After giving a brief description of the evolution of CCP4 over the last 30 years, an overview of the current suite is given.

Abstract: While detailed descriptions are given in the accompanying articles, here it is shown how the individual programs contribute to a complete software package.


Histone Demethylase Expression Enhances Human Somatic Cell Nuclear Transfer Efficiency and Promotes Derivation of Pluripotent Stem Cells.

Abstract: The extremely low efficiency of human embryonic stem cell (hESC) derivation using somatic cell nuclear transfer (SCNT) limits its potential application.

Abstract: Blastocyst formation from human SCNT embryos occurs at a low rate and with only some oocyte donors.

Abstract: We previously showed in mice that reduction of histone H3 lysine 9 trimethylation (H3K9me3) through ectopic expression of the H3K9me3 demethylase Kdm4d greatly improves SCNT embryo development.

Abstract: Here we show that overexpression of a related H3K9me3 demethylase KDM4A improves human SCNT, and that, as in mice, H3K9me3 in the human somatic cell genome is an SCNT reprogramming barrier.

Abstract: Overexpression of KDM4A significantly improves the blastocyst formation rate in human SCNT embryos by facilitating transcriptional reprogramming, allowing efficient derivation of SCNT-derived ESCs using adult Age-related Macular Degeneration (AMD) patient somatic nuclei donors.

Abstract: This conserved mechanistic insight has potential applications for improving SCNT in a variety of contexts, including regenerative medicine.


Interface Contractility between Differently Fated Cells Drives Cell Elimination and Cyst Formation

Abstract: Although cellular tumor-suppression mechanisms are widely studied, little is known about mechanisms that act at the level of tissues to suppress the occurrence of aberrant cells in epithelia.

Abstract: We find that ectopic expression of transcription factors that specify cell fates causes abnormal epithelial cysts in Drosophila imaginal discs.

Abstract: Cysts do not form cell autonomously but result from the juxtaposition of two cell populations with divergent fates.

Abstract: Juxtaposition of wild-type and aberrantly specified cells induces enrichment of actomyosin at their entire shared interface, both at adherens junctions as well as along basolateral interfaces.

Abstract: Experimental validation of 3D vertex model simulations demonstrates that enhanced interface contractility is sufficient to explain many morphogenetic behaviors, which depend on cell cluster size.

Abstract: These range from cyst formation by intermediate-sized clusters to segregation of large cell populations by formation of smooth boundaries or apical constriction in small groups of cells.

Abstract: In addition, we find that single cells experiencing lateral interface contractility are eliminated from tissues by apoptosis.

Abstract: Cysts, which disrupt epithelial continuity, form when elimination of single, aberrantly specified cells fails and cells proliferate to intermediate cell cluster sizes.

Abstract: Thus, increased interface contractility functions as error correction mechanism eliminating single aberrant cells from tissues, but failure leads to the formation of large, potentially disease-promoting cysts.

Abstract: Our results provide a novel perspective on morphogenetic mechanisms, which arise from cell-fate heterogeneities within tissues and maintain or disrupt epithelial homeostasis.


Resonating circadian clocks enhance fitness in cyanobacteria.

Abstract: In some organisms longevity, growth, and developmental rate are improved when they are maintained on a light/dark cycle, the period of which "resonates" optimally with the period of the endogenous circadian clock.

Abstract: However, to our knowledge no studies have demonstrated that reproductive fitness per se is improved by resonance between the endogenous clock and the environmental cycle.

Abstract: We tested the adaptive significance of circadian programming by measuring the relative fitness under competition between various strains of cyanobacteria expressing different circadian periods.

Abstract: Strains that had a circadian period similar to that of the light/dark cycle were favored under competition in a manner that indicates the action of soft selection.


Stromal Fibroblasts Present in Invasive Human Breast Carcinomas Promote Tumor Growth and Angiogenesis through Elevated SDF-1/CXCL12 Secretion

Abstract: Fibroblasts often constitute the majority of the stromal cells within a breast carcinoma, yet the functional contributions of these cells to tumorigenesis are poorly understood.

Abstract: Using a coimplantation tumor xenograft model, we demonstrate that carcinoma-associated fibroblasts (CAFs) extracted from human breast carcinomas promote the growth of admixed breast carcinoma cells significantly more than do normal mammary fibroblasts derived from the same patients.

Abstract: The CAFs, which exhibit the traits of myofibroblasts, play a central role in promoting the growth of tumor cells through their ability to secrete stromal cell-derived factor 1 (SDF-1); CAFs promote angiogenesis by recruiting endothelial progenitor cells (EPCs) into carcinomas, an effect mediated in part by SDF-1.

Abstract: CAF-secreted SDF-1 also stimulates tumor growth directly, acting through the cognate receptor, CXCR4, which is expressed by carcinoma cells.

Abstract: Our findings indicate that fibroblasts within invasive breast carcinomas contribute to tumor promotion in large part through the secretion of SDF-1.


Embryonic Origin of Postnatal Neural Stem Cells

Abstract: Adult neural stem/progenitor (B1) cells within the walls of the lateral ventricles generate different types of neurons for the olfactory bulb (OB).

Abstract: The location of B1 cells determines the types of OB neurons they generate.

Abstract: Here we show that the majority of mouse B1 cell precursors are produced between embryonic days (E) 13.5 and 15.5 and remain largely quiescent until they become reactivated postnatally.

Abstract: Using a retroviral library carrying over 100,000 genetic tags, we found that B1 cells share a common progenitor with embryonic cells of the cortex, striatum, and septum, but this lineage relationship is lost before E15.5.

Abstract: The regional specification of B1 cells is evident as early as E11.5 and is spatially linked to the production of neurons that populate different areas of the forebrain.

Abstract: This study reveals an early embryonic regional specification of postnatal neural stem cells and the lineage relationship between them and embryonic progenitor cells.


The Opa1-Dependent Mitochondrial Cristae Remodeling Pathway Controls Atrophic, Apoptotic, and Ischemic Tissue Damage

Abstract: Mitochondrial morphological and ultrastructural changes occur during apoptosis and autophagy, but whether they are relevant in vivo for tissue response to damage is unclear.

Abstract: Here we investigate the role of the optic atrophy 1 (OPA1)-dependent cristae remodeling pathway in vivo and provide evidence that it regulates the response of multiple tissues to apoptotic, necrotic, and atrophic stimuli.

Abstract: Genetic inhibition of the cristae remodeling pathway in vivo does not affect development, but protects mice from denervation-induced muscular atrophy, ischemic heart and brain damage, as well as hepatocellular apoptosis.

Abstract: Mechanistically, OPA1-dependent mitochondrial cristae stabilization increases mitochondrial respiratory efficiency and blunts mitochondrial dysfunction, cytochrome c release, and reactive oxygen species production.

Abstract: Our results indicate that the OPA1-dependent cristae remodeling pathway is a fundamental, targetable determinant of tissue damage in vivo.


First Proposal of Minimum Information About a Cellular Assay for Regenerative Medicine

Abstract: : Advances in stem cell research have triggered scores of studies in regenerative medicine in a large number of institutions and companies around the world.

Abstract: However, reproducibility and data exchange among laboratories or cell banks are constrained by the lack of a standardized format for experiments.

Abstract: To enhance information flow in stem cell and derivative cell research, here we propose a minimum information standard to describe cellular assay data to facilitate practical regenerative medicine.

Abstract: Based on the existing Minimum Information About a Cellular Assay, we developed Minimum Information About a Cellular Assay for Regenerative Medicine (MIACARM), which allows for the description of advanced cellular experiments with defined taxonomy of human cell types.

Abstract: By using controlled terms, such as ontologies, MIACARM will provide a platform for cellular assay data exchange among cell banks or registries that have been established at more than 20 sites in the world.

Abstract: SIGNIFICANCE Currently, there are more than 20 human cell information storage sites around the world.

Abstract: However, reproducibility and data exchange among different laboratories or cell information providers are usually inadequate or nonexistent because of the lack of a standardized format for experiments.

Abstract: This study, which is the fruit of collaborative work by scientists at stem cell banks and cellular information registries worldwide, including those in the U.S., the U.K., Europe, and Japan, proposes new minimum information guidelines, Minimum Information About a Cellular Assay for Regenerative Medicine (MIACARM), for cellular assay data deposition.

Abstract: MIACARM is intended to promote data exchange and facilitation of practical regenerative medicine.


The PAR proteins: fundamental players in animal cell polarization.

Abstract: The par genes were discovered in genetic screens for regulators of cytoplasmic partitioning in the early embryo of C. elegans, and encode six different proteins required for asymmetric cell division by the worm zygote.

Abstract: Some of the PAR proteins are localized asymmetrically and form physical complexes with one another.

Abstract: Strikingly, the PAR proteins have been found to regulate cell polarization in many different contexts in diverse animals, suggesting they form part of an ancient and fundamental mechanism for cell polarization.

Abstract: Although the picture of how the PAR proteins function remains incomplete, cell biology and biochemistry are beginning to explain how PAR proteins polarize cells.


Prions Adhere to Soil Minerals and Remain Infectious

Abstract: An unidentified environmental reservoir of infectivity contributes to the natural transmission of prion diseases (transmissible spongiform encephalopathies [TSEs]) in sheep, deer, and elk.

Abstract: Prion infectivity may enter soil environments via shedding from diseased animals and decomposition of infected carcasses.

Abstract: Burial of TSE-infected cattle, sheep, and deer as a means of disposal has resulted in unintentional introduction of prions into subsurface environments.

Abstract: We examined the potential for soil to serve as a TSE reservoir by studying the interaction of the disease-associated prion protein (PrPSc) with common soil minerals.

Abstract: In this study, we demonstrated substantial PrPSc adsorption to two clay minerals, quartz, and four whole soil samples.

Abstract: We quantified the PrPSc-binding capacities of each mineral.

Abstract: Furthermore, we observed that PrPSc desorbed from montmorillonite clay was cleaved at an N-terminal site and the interaction between PrPSc and Mte was strong, making desorption of the protein difficult.

Abstract: Despite cleavage and avid binding, PrPSc bound to Mte remained infectious.

Abstract: Results from our study suggest that PrPSc released into soil environments may be preserved in a bioavailable form, perpetuating prion disease epizootics and exposing other species to the infectious agent.


Regulated Accumulation of Desmosterol Integrates Macrophage Lipid Metabolism and Inflammatory Responses

Abstract: Inflammation and macrophage foam cells are characteristic features of atherosclerotic lesions, but the mechanisms linking cholesterol accumulation to inflammation and LXR-dependent response pathways are poorly understood.

Abstract: To investigate this relationship, we utilized lipidomic and transcriptomic methods to evaluate the effect of diet and LDL receptor genotype on macrophage foam cell formation within the peritoneal cavities of mice.

Abstract: Foam cell formation was associated with significant changes in hundreds of lipid species and unexpected suppression, rather than activation, of inflammatory gene expression.

Abstract: We provide evidence that regulated accumulation of desmosterol underlies many of the homeostatic responses, including activation of LXR target genes, inhibition of SREBP target genes, selective reprogramming of fatty acid metabolism, and suppression of inflammatory-response genes, observed in macrophage foam cells.

Abstract: These observations suggest that macrophage activation in atherosclerotic lesions results from extrinsic, proinflammatory signals generated within the artery wall that suppress homeostatic and anti-inflammatory functions of desmosterol.


A JOURNAL OF NEUROLOGY

Abstract: Brain glial cells, five times more prevalent than neurons, have recently received attention for their potential involvement in epileptic seizures.

Abstract: Microglia and astrocytes, associated with inflammatory innate immune responses, are responsible for surveillance of brain damage that frequently results in seizures.

Abstract: Thus, an intriguing suggestion has been put forward that seizures may be facilitated and perhaps triggered by brain immune responses.

Abstract: Indeed, recent evidence strongly implicates innate immune responses in lowering seizure threshold in experimental models of epilepsy, yet, there is no proof that they can play an independent role in initiating seizures in vivo.

Abstract: Here, we show that cortical innate immune responses alone produce profound increases of brain excitability resulting in focal seizures.

Abstract: We found that cortical application of lipopolysaccharide, binding to toll-like receptor 4 (TLR4), triples evoked field potential amplitudes and produces focal epileptiform discharges.

Abstract: These effects are prevented by pre-application of interleukin-1 receptor antagonist.

Abstract: Our results demonstrate how the innate immune response may participate in acute seizures, increasing neuronal excitability through interleukin-1 release in response to TLR4 detection of the danger signals associated with infections of the central nervous system and with brain injury.

Abstract: These results suggest an important role of innate immunity in epileptogenesis and focus on glial inhibition, through pharmacological blockade of TLR4 and the pro-inflammatory mediators released by activated glia, in the study and treatment of seizure disorders in humans.


Sequential domain assembly of ribosomal protein S3 drives 40S subunit maturation.

Abstract: Eukaryotic ribosomes assemble by association of ribosomal RNA with ribosomal proteins into nuclear precursor particles, which undergo a complex maturation pathway coordinated by non-ribosomal assembly factors.

Abstract: Here, we provide functional insights into how successive structural re-arrangements in ribosomal protein S3 promote maturation of the 40S ribosomal subunit.

Abstract: We show that S3 dimerizes and is imported into the nucleus with its N-domain in a rotated conformation and associated with the chaperone Yar1.

Abstract: Initial assembly of S3 with 40S precursors occurs via its C-domain, while the N-domain protrudes from the 40S surface.

Abstract: Yar1 is replaced by the assembly factor Ltv1, thereby fixing the S3 N-domain in the rotated orientation and preventing its 40S association.

Abstract: Finally, Ltv1 release, triggered by phosphorylation, and flipping of the S3 N-domain into its final position results in the stable integration of S3.

Abstract: Such a stepwise assembly may represent a new paradigm for the incorporation of ribosomal proteins.


Assaying proliferation and differentiation capacity of stem cells using disaggregated adult mouse epidermis

Abstract: In this protocol, we describe how to isolate keratinocytes from adult mouse epidermis, fractionate them into different sub-populations on the basis of cell surface markers and examine their function in an in vivo skin reconstitution assay with disaggregated neonatal dermal cells.

Abstract: We also describe how the isolated keratinocytes can be subjected to clonal analysis in vitro and in vivo and how to enrich for hair follicle-inducing dermal papilla cells in the dermal preparation.

Abstract: Using these approaches, it is possible to compare the capacity of different populations of adult epidermal stem cells to proliferate and to generate progeny that differentiate along the different epidermal lineages.

Abstract: Isolating, fractionating and grafting cells for the skin reconstitution assay is normally spread over 2 d. Clonal growth in culture is assessed after 14 d, while evaluation of the grafts is carried out after 4–5 weeks.


Telomerase-negative immortalized human cells contain a novel type of promyelocytic leukemia (PML) body.

Abstract: Telomerase-negative immortalized human cells maintain their telomeres by a mechanism known as alternative lengthening of telomeres (ALT).

Abstract: We report here that ALT cells contain a novel promyelocytic leukemia (PML) body (ALT-associated PML body, APB).

Abstract: APBs are large donut-shaped nuclear structures containing PML protein, telomeric DNA, and the telomere binding proteins human telomere repeat binding factors 1 and 2.

Abstract: Immunostaining showed that APBs also contain replication factor A, RAD51, and RAD52, proteins involved in DNA synthesis and recombination.

Abstract: During immortalization, APBs appeared at exactly the same time as activation of ALT.

Abstract: APBs were found in ALT tumors and cell lines but not in mortal cell strains or in telomerase-positive cell lines or tumors.


Prediction of coronary artery disease by a systemic atherosclerosis score index derived from whole-body MR angiography

Abstract: BACKGROUND Whole-body magnetic resonance angiography (WB-MRA) has shown its potential for the non-invasive assessment of nearly the entire arterial vasculature within one examination.

Abstract: Since the presence of extra-cardiac atherosclerosis is associated with an increased risk of coronary events, our goal was to establish the relationship between WB-MRA findings, including a systemic atherosclerosis score index, and the presence of significant coronary artery disease (CAD).

Abstract: METHODS WB-MRA was performed on a 1.5T scanner in 50 patients scheduled to undergo elective cardiac catheterization for suspected CAD.

Abstract: In each patient, 40 extra-cardiac vessel segments were evaluated and assigned scores according to their luminal narrowing.

Abstract: The atherosclerosis score index (ASI) was generated as the ratio of summed scores to analyzable segments.

Abstract: RESULTS ASI was higher in patients with significant (> 50% stenosis) CAD (n = 27) vs. patients without CAD (n = 22; 1.56 vs. 1.28, p = 0.004).

Abstract: ASI correlated with PROCAM (R = 0.57, p < 0.001) and Framingham (R = 0.36, p = 0.01) risk scores as estimates of the 10-year risk of coronary events.

Abstract: A ROC derived ASI of > 1.54 predicted significant CAD with a sensitivity of 59%, specificity of 86% and a positive predictive value of 84%.

Abstract: Logistic regression revealed ASI > 1.54 as the strongest independent predictor for CAD with a 11-fold increase in likelihood to suffer from significant coronary disease.

Abstract: On the contrary, while 15/27 (55%) of patients with CAD exhibited at least one extra-cardiac stenosis > 50%, only 3/22 (14%) of those patients without CAD did (p = 0.003).

Abstract: The likelihood for an extra-cardiac stenosis when CAD is present differed between vascular territories and ranged from 15% for a carotid stenosis to 44% for a stenosis in the lower extremities.

Abstract: CONCLUSION This study provides important new evidence for the close association of extra-cardiac and coronary atherosclerosis.

Abstract: The novel findings that a WB-MRA derived systemic atherosclerosis score index is not only associated with established cardiovascular risk scores but is also predictive of significant CAD suggest its potential prognostic implications and underline the importance to screen for coronary disease in patients with extra-cardiac manifestations of atherosclerosis.


Analysis of re-replication from deregulated origin licensing by DNA fiber spreading

Abstract: A major challenge each human cell-division cycle is to ensure that DNA replication origins do not initiate more than once, a phenomenon known as re-replication.

Abstract: Acute deregulation of replication control ultimately causes extensive DNA damage, cell-cycle checkpoint activation and cell death whereas moderate deregulation promotes genome instability and tumorigenesis.

Abstract: In the absence of detectable increases in cellular DNA content however, it has been difficult to directly demonstrate re-replication or to determine if the ability to re-replicate is restricted to a particular cell-cycle phase.

Abstract: Using an adaptation of DNA fiber spreading we report the direct detection of re-replication on single DNA molecules from human chromosomes.

Abstract: Using this method we demonstrate substantial re-replication within 1 h of S phase entry in cells overproducing the replication factor, Cdt1.

Abstract: Moreover, a comparison of the HeLa cancer cell line to untransformed fibroblasts suggests that HeLa cells produce replication signals consistent with low-level re-replication in otherwise unperturbed cell cycles.

Abstract: Re-replication after depletion of the Cdt1 inhibitor, geminin, in an untransformed fibroblast cell line is undetectable by standard assays but readily quantifiable by DNA fiber spreading analysis.

Abstract: Direct evaluation of re-replicated DNA molecules will promote increased understanding of events that promote or perturb genome stability.


Liver receptor homolog-1 is essential for pregnancy

Abstract: Successful pregnancy requires coordination of an array of signals and factors from multiple tissues.

Abstract: One such element, liver receptor homolog-1 (Lrh-1), is an orphan nuclear receptor that regulates metabolism and hormone synthesis.

Abstract: It is strongly expressed in granulosa cells of ovarian follicles and in the corpus luteum of rodents and humans.

Abstract: Germline ablation of Nr5a2 (also called Lrh-1), the gene coding for Lrh-1, in mice is embryonically lethal at gastrulation.

Abstract: Depletion of Lrh-1 in the ovarian follicle shows that it regulates genes required for both steroid synthesis and ovulation.

Abstract: To study the effects of Lrh-1 on mouse gestation, we genetically disrupted its expression in the corpus luteum, resulting in luteal insufficiency.

Abstract: Hormone replacement permitted embryo implantation but was followed by gestational failure with impaired endometrial decidualization, compromised placental formation, fetal growth retardation and fetal death.

Abstract: Lrh-1 is also expressed in the mouse and human endometrium, and in a primary culture of human endometrial stromal cells, reduction of NR5A2 transcript abundance by RNA interference abrogated decidualization.

Abstract: These findings show that Lrh-1 is necessary for maintenance of the corpus luteum, for promotion of decidualization and for formation of the placenta.

Abstract: It therefore has multiple, indispensible roles in establishing and sustaining pregnancy.


Transcriptional Signature and Memory Retention of Human-Induced Pluripotent Stem Cells

Abstract: Genetic reprogramming of somatic cells to a pluripotent state (induced pluripotent stem cells or iPSCs) by over-expression of specific genes has been accomplished using mouse and human cells.

Abstract: However, it is still unclear how similar human iPSCs are to human Embryonic Stem Cells (hESCs).

Abstract: Here, we describe the transcriptional profile of human iPSCs generated without viral vectors or genomic insertions, revealing that these cells are in general similar to hESCs but with significant differences.

Abstract: For the generation of human iPSCs without viral vectors or genomic insertions, pluripotent factors Oct4 and Nanog were cloned in episomal vectors and transfected into human fetal neural progenitor cells.

Abstract: The transient expression of these two factors, or from Oct4 alone, resulted in efficient generation of human iPSCs.

Abstract: The reprogramming strategy described here revealed a potential transcriptional signature for human iPSCs yet retaining the gene expression of donor cells in human reprogrammed cells free of viral and transgene interference.

Abstract: Moreover, the episomal reprogramming strategy represents a safe way to generate human iPSCs for clinical purposes and basic research.


A CXCL1 Paracrine Network Links Cancer Chemoresistance and Metastasis

Abstract: Metastasis and chemoresistance in cancer are linked phenomena, but the molecular basis for this link is unknown.

Abstract: We uncovered a network of paracrine signals between carcinoma, myeloid, and endothelial cells that drives both processes in breast cancer.

Abstract: Cancer cells that overexpress CXCL1 and 2 by transcriptional hyperactivation or 4q21 amplification are primed for survival in metastatic sites.

Abstract: CXCL1/2 attract CD11b(+)Gr1(+) myeloid cells into the tumor, which produce chemokines including S100A8/9 that enhance cancer cell survival.

Abstract: Although chemotherapeutic agents kill cancer cells, these treatments trigger a parallel stromal reaction leading to TNF-α production by endothelial and other stromal cells.

Abstract: TNF-α via NF-kB heightens the CXCL1/2 expression in cancer cells, thus amplifying the CXCL1/2-S100A8/9 loop and causing chemoresistance.

Abstract: CXCR2 blockers break this cycle, augmenting the efficacy of chemotherapy against breast tumors and particularly against metastasis.

Abstract: This network of endothelial-carcinoma-myeloid signaling interactions provides a mechanism linking chemoresistance and metastasis, with opportunities for intervention.


Prophylaxis of postoperative vomiting in children undergoing tonsillectomy: a systematic review and meta-analysis.

Abstract: Postoperative vomiting (POV) remains one of the commonest causes of significant morbidity after tonsillectomy in children.

Abstract: A variety of prophylactic anti-emetic interventions have been reported, but there has only been a limited systematic review in this patient group.

Abstract: A systematic search was performed by using Cochrane Controlled Trials Register, MEDLINE and EMBASE to identify double-blind, randomized, placebo-controlled trials of prophylactic anti-emetic interventions in children undergoing tonsillectomy, with or without adenoidectomy.

Abstract: The outcome of interest was POV in the first 24 h. Summary estimates of the effect of each prophylactic anti-emetic strategy were derived using fixed effect meta-analysis.

Abstract: Where appropriate, dose-response effects were estimated using logistic regression and 22 articles were identified.

Abstract: Good evidence was found for the prophylactic anti-emetic effect of dexamethasone [odds ratio (OR) 0.23, 95% CI 0.16-0.33], and the serotinergic antagonists ondansetron (OR 0.36, 95% CI 0.29-0.46), granisetron (OR 0.11, 95% CI 0.06-0.19), tropisetron (OR 0.15, 95% CI 0.06-0.35) and dolasetron (OR 0.25, 95% CI 0.1-0.59).

Abstract: Metoclopramide was also found to be efficacious (OR 0.51, 95% CI 0.34-0.77).

Abstract: There is not sufficient evidence to suggest that dimenhydrinate, perphenazine or droperidol, in the doses studied, are efficacious, nor were gastric aspiration or acupuncture.

Abstract: In conclusion, dexamethasone and the anti-serotinergic agents appear to be the most effective agents for the prophylaxis for POV in children undergoing tonsillectomy.


A method of comparing the areas under receiver operating characteristic curves derived from the same cases.

Abstract: Receiver operating characteristic (ROC) curves are used to describe and compare the performance of diagnostic technology and diagnostic algorithms.

Abstract: This paper refines the statistical comparison of the areas under two ROC curves derived from the same set of patients by taking into account the correlation between the areas that is induced by the paired nature of the data.

Abstract: The correspondence between the area under an ROC curve and the Wilcoxon statistic is used and underlying Gaussian distributions (binormal) are assumed to provide a table that converts the observed correlations in paired ratings of images into a correlation between the two ROC areas.

Abstract: This between-area correlation can be used to reduce the standard error (uncertainty) about the observed difference in areas.

Abstract: This correction for pairing, analogous to that used in the paired t-test, can produce a considerable increase in the statistical sensitivity (power) of the comparison.

Abstract: For studies involving multiple readers, this method provides a measure of a component of the sampling variation that is otherwise difficult to obtain.


Transient inactivation of Rb and ARF yields regenerative cells from postmitotic mammalian muscle.

Abstract: An outstanding biological question is why tissue regeneration in mammals is limited, whereas urodele amphibians and teleost fish regenerate major structures, largely by cell cycle reentry.

Abstract: Upon inactivation of Rb, proliferation of postmitotic urodele skeletal muscle is induced, whereas in mammalian muscle this mechanism does not exist.

Abstract: We postulated that a tumor suppressor present in mammals but absent in regenerative vertebrates, the Ink4a product ARF (alternative reading frame), is a regeneration suppressor.

Abstract: Concomitant inactivation of Arf and Rb led to mammalian muscle cell cycle reentry, loss of differentiation properties, and upregulation of cytokinetic machinery.

Abstract: Single postmitotic myocytes were isolated by laser micro-dissection-catapulting, and transient suppression of Arf and Rb yielded myoblast colonies that retained the ability to differentiate and fuse into myofibers upon transplantation in vivo.

Abstract: These results show that differentiation of mammalian cells is reversed by inactivation of Arf and Rb and support the hypothesis that Arf evolved at the expense of regeneration.


CENP-B Controls Centromere Formation Depending on the Chromatin Context

Abstract: The centromere is a chromatin region that serves as the spindle attachment point and directs accurate inheritance of eukaryotic chromosomes during cell divisions.

Abstract: However, the mechanism by which the centromere assembles and stabilizes at a specific genomic region is not clear.

Abstract: The de novo formation of a human/mammalian artificial chromosome (HAC/MAC) with a functional centromere assembly requires the presence of alpha-satellite DNA containing binding motifs for the centromeric CENP-B protein.

Abstract: We demonstrate here that de novo centromere assembly on HAC/MAC is dependent on CENP-B. In contrast, centromere formation is suppressed in cells expressing CENP-B when alpha-satellite DNA was integrated into a chromosomal site.

Abstract: Remarkably, on those integration sites CENP-B enhances histone H3-K9 trimethylation and DNA methylation, thereby stimulating heterochromatin formation.

Abstract: Thus, we propose that CENP-B plays a dual role in centromere formation, ensuring de novo formation on DNA lacking a functional centromere but preventing the formation of excess centromeres on chromosomes.


Improved tools for biological sequence comparison.

Abstract: We have developed three computer programs for comparisons of protein and DNA sequences.

Abstract: They can be used to search sequence data bases, evaluate similarity scores, and identify periodic structures based on local sequence similarity.

Abstract: The FASTA program is a more sensitive derivative of the FASTP program, which can be used to search protein or DNA sequence data bases and can compare a protein sequence to a DNA sequence data base by translating the DNA data base as it is searched.

Abstract: FASTA includes an additional step in the calculation of the initial pairwise similarity score that allows multiple regions of similarity to be joined to increase the score of related sequences.

Abstract: The RDF2 program can be used to evaluate the significance of similarity scores using a shuffling method that preserves local sequence composition.

Abstract: The LFASTA program can display all the regions of local similarity between two sequences with scores greater than a threshold, using the same scoring parameters and a similar alignment algorithm; these local similarities can be displayed as a "graphic matrix" plot or as individual alignments.

Abstract: In addition, these programs have been generalized to allow comparison of DNA or protein sequences based on a variety of alternative scoring matrices.


The adverse consequences of unmet need among older persons living in the community: dual-eligible versus Medicare-only beneficiaries.

Abstract: OBJECTIVE Our objective is to estimate and compare the prevalence of selected adverse consequences associated with unmet need for assistance among a socioeconomically and medically vulnerable subgroup of the older adult population, those who are dually eligible for Medicare and Medicaid, with those eligible for Medicare only.

Abstract: METHOD Using data from the National Health and Aging Trends Study (NHATS), a representative survey of the older Medicare population, we calculated the prevalence of disability-related need for assistance with self-care, household tasks, and mobility activities and the prevalence of adverse consequences of unmet need by dually eligible and Medicare only status.

Abstract: RESULTS Over 2 million community-dwelling older persons experienced an adverse consequence due to unmet need for assistance with self-care (e.g., soiled their clothes), over 2 million experienced adverse consequences due to unmet need for assistance with household tasks (e.g., went without groceries), and over 3 million persons experienced at least one adverse consequence of unmet need for assistance with mobility-related activities (e.g., had to stay in bed) in the month prior to the NHATS interview.

Abstract: Dually eligible persons experienced higher rates of 6 of the 11 adverse consequences studied and were more likely to have at least one adverse consequence in all 3 domains than others.

Abstract: DISCUSSION Several care models are emerging with the goal of integrating medical care, behavioral health, and long-term services for the dual eligible population.

Abstract: Indicators of adverse consequences of unmet need could be used to monitor the quality and adequacy of such care systems.


Supplementary information: ‘MesoRD User’s Guide ’ and other

Abstract: UNLABELLED MesoRD is a tool for stochastic simulation of chemical reactions and diffusion.

Abstract: In particular, it is an implementation of the next subvolume method, which is an exact method to simulate the Markov process corresponding to the reaction-diffusion master equation.

Abstract: AVAILABILITY MesoRD is free software, written in C++ and licensed under the GNU general public license (GPL).

Abstract: MesoRD runs on Linux, Mac OS X, NetBSD, Solaris and Windows XP.

Abstract: It can be downloaded from http://mesord.sourceforge.net.

Abstract: CONTACT johan.elf@icm.uu.se; johan.hattne@embl-hamburg.de   SUPPLEMENTARY INFORMATION 'MesoRD User's Guide' and other documents are available at http://mesord.sourceforge.net.


Functional anatomy of T cell activation and synapse formation.

Abstract: T cell activation and function require a structured engagement of antigen-presenting cells.

Abstract: These cell contacts are characterized by two distinct dynamics in vivo: transient contacts resulting from promigratory junctions called immunological kinapses or prolonged contacts from stable junctions called immunological synapses.

Abstract: Kinapses operate in the steady state to allow referencing to self-peptide-MHC (pMHC) and searching for pathogen-derived pMHC.

Abstract: Synapses are induced by T cell receptor (TCR) interactions with agonist pMHC under specific conditions and correlate with robust immune responses that generate effector and memory T cells.

Abstract: High-resolution imaging has revealed that the synapse is highly coordinated, integrating cell adhesion, TCR recognition of pMHC complexes, and an array of activating and inhibitory ligands to promote or prevent T cell signaling.

Abstract: In this review, we examine the molecular components, geometry, and timing underlying kinapses and synapses.

Abstract: We integrate recent molecular and physiological data to provide a synthesis and suggest ways forward.


Modulation of Replicative Lifespan in Cryptococcus neoformans: Implications for Virulence

Abstract: The fungal pathogen, Cryptococcus neoformans, has been shown to undergo replicative aging.

Abstract: Old cells are characterized by advanced generational age and phenotypic changes that appear to mediate enhanced resistance to host and antifungal-based killing.

Abstract: As a consequence of this age-associated resilience, old cells accumulate during chronic infection.

Abstract: Based on these findings, we hypothesized that shifting the generational age of a pathogenic yeast population would alter its vulnerability to the host and affect its virulence.

Abstract: SIR2 is a well-conserved histone deacetylase, and a pivotal target for the development of anti-aging drugs.

Abstract: We tested its effect on C. neoformans' replicative lifespan (RLS).

Abstract: First, a mutant C. neoformans strain (sir2Δ) was generated, and confirmed a predicted shortened RLS in sir2Δ cells consistent with its known role in aging.

Abstract: Next, RLS analysis showed that treatment of C. neoformans with Sir2p-agonists resulted in a significantly prolonged RLS, whereas treatment with a Sir2p-antagonist shortened RLS.

Abstract: RLS modulating effects were dependent on SIR2 and not observed in sir2Δ cells.

Abstract: Because SIR2 loss resulted in a slightly impaired fitness, effects of genetic RLS modulation on virulence could not be compared with wild type cells.

Abstract: Instead we chose to chemically modulate RLS, and investigated the effect of Sir2p modulating drugs on C. neoformans cells in a Galleria mellonella infection model.

Abstract: Consistent with our hypothesis that shifts in the generational age of the infecting yeast population alters its vulnerability to host cells, we observed decreased virulence of C. neoformans in the Galleria host when RLS was prolonged by treatment with Sir2p agonists.

Abstract: In contrast, treatment with a Sir2p antagonist, which shortens RLS enhanced virulence in Galleria.

Abstract: In addition, combination of Sir2p agonists with antifungal therapy enhanced the antifungal's effect.

Abstract: Importantly, no difference in virulence was observed with drug treatment when sir2Δ cells were used for infection, which confirmed target specificity and ruled out non-specific effects of the drugs on the Galleria host.

Abstract: Thus, this study suggests that RLS modulating drugs, such as Sir2p agonists, shift lifespan and vulnerability of the fungal population, and should be further investigated as a potential class of novel antifungal drug targets that can enhance antifungal efficacy.


Pf4-Cre transgenic mice allow the generation of lineage-restricted gene knockouts for studying megakaryocyte and platelet function in vivo.

Abstract: To generate transgenic mice that express Cre-recombinase exclusively in the megakaryocytic lineage, we modified a mouse bacterial artificial chromosome (BAC) clone by homologous recombination and replaced the first exon of the platelet factor 4 (Pf4), also called CXCL4, with a codon-improved Cre cDNA.

Abstract: Several strains expressing the transgene were obtained and one strain, Q3, was studied in detail.

Abstract: Crossing Q3 mice with the ROSA26-lacZ reporter strain showed that Cre-recombinase activity was confined to megakaryocytes.

Abstract: These results were further verified by crossing the Q3 mice with a strain containing loxP-flanked integrin beta1.

Abstract: Excision of this conditional allele in megakaryocytes was complete at the DNA level, and platelets were virtually devoid of the integrin beta1 protein.

Abstract: The Pf4-Cre transgenic strain will be a valuable tool to study megakaryopoiesis, platelet formation, and platelet function.


LIN-44/Wnt Directs Dendrite Outgrowth through LIN-17/Frizzled in C. elegans Neurons

Abstract: Nervous system function requires proper development of two functional and morphological domains of neurons, axons and dendrites.

Abstract: Although both these domains are equally important for signal transmission, our understanding of dendrite development remains relatively poor.

Abstract: Here, we show that in C. elegans the Wnt ligand, LIN-44, and its Frizzled receptor, LIN-17, regulate dendrite development of the PQR oxygen sensory neuron.

Abstract: In lin-44 and lin-17 mutants, PQR dendrites fail to form, display stunted growth, or are misrouted.

Abstract: Manipulation of temporal and spatial expression of LIN-44, combined with cell-ablation experiments, indicates that this molecule is patterned during embryogenesis and acts as an attractive cue to define the site from which the dendrite emerges.

Abstract: Genetic interaction between lin-44 and lin-17 suggests that the LIN-44 signal is transmitted through the LIN-17 receptor, which acts cell autonomously in PQR.

Abstract: Furthermore, we provide evidence that LIN-17 interacts with another Wnt molecule, EGL-20, and functions in parallel to MIG-1/Frizzled in this process.

Abstract: Taken together, our results reveal a crucial role for Wnt and Frizzled molecules in regulating dendrite development in vivo.


Why do general practitioners prescribe antibiotics for upper respiratory tract infections to meet patient expectations: a mixed methods study

Abstract: OBJECTIVES To describe the role patient expectations play in general practitioners (GPs) antibiotic prescribing for upper respiratory tract infections (URTI).

Abstract: METHODS Concurrent explanatory mixed methods approach using a cross-sectional survey and semistructured interviews.

Abstract: SETTINGS Primary care GPs in Australia.

Abstract: PARTICIPANTS 584 GPs (response rate of 23.6%) completed the cross-sectional survey.

Abstract: 32 GPs were interviewed individually.

Abstract: OUTCOME MEASURE Prescribing of antibiotics for URTI.

Abstract: RESULTS More than half the GP respondents to the survey in Australia self-reported that they would prescribe antibiotics for an URTI to meet patient expectations.

Abstract: Our qualitative findings suggest that 'patient expectations' may be the main reason given for inappropriate prescribing, but it is an all-encompassing phrase that includes other reasons.

Abstract: These include limited time, poor doctor-patient communication and diagnostic uncertainty.

Abstract: We have identified three role archetypes to explain the behaviour of GPs in reference to antibiotic prescribing for URTIs.

Abstract: The main themes emerging from the qualitative component was that many GPs did not think that antibiotic prescribing in primary care was responsible for the development of antibiotic resistance nor that their individual prescribing would make any difference in light of other bigger issues like hospital prescribing or veterinary use.

Abstract: For them, there were negligible negative consequences from their inappropriate prescribing.

Abstract: CONCLUSIONS There is a need to increase awareness of the scope and magnitude of antibiotic resistance and the role primary care prescribing plays, and of the contribution of individual prescribing decisions to the problem of antibiotic resistance.


Migraine and cardiovascular disease: systematic review and meta-analysis.

Abstract: OBJECTIVE To evaluate the association between migraine and cardiovascular disease, including stroke, myocardial infarction, and death due to cardiovascular disease.

Abstract: DESIGN Systematic review and meta-analysis.

Abstract: DATA SOURCES Electronic databases (PubMed, Embase, Cochrane Library) and reference lists of included studies and reviews published until January 2009.

Abstract: Selection criteria Case-control and cohort studies investigating the association between any migraine or specific migraine subtypes and cardiovascular disease.

Abstract: Review methods Two investigators independently assessed eligibility of identified studies in a two step approach.

Abstract: Disagreements were resolved by consensus.

Abstract: Studies were grouped according to a priori categories on migraine and cardiovascular disease.

Abstract: DATA EXTRACTION Two investigators extracted data.

Abstract: Pooled relative risks and 95% confidence intervals were calculated.

Abstract: RESULTS Studies were heterogeneous for participant characteristics and definition of cardiovascular disease.

Abstract: Nine studies investigated the association between any migraine and ischaemic stroke (pooled relative risk 1.73, 95% confidence interval 1.31 to 2.29).

Abstract: Additional analyses indicated a significantly higher risk among people who had migraine with aura (2.16, 1.53 to 3.03) compared with people who had migraine without aura (1.23, 0.90 to 1.69; meta-regression for aura status P=0.02).

Abstract: Furthermore, results suggested a greater risk among women (2.08, 1.13 to 3.84) compared with men (1.37, 0.89 to 2.11).

Abstract: Age less than 45 years, smoking, and oral contraceptive use further increased the risk.

Abstract: Eight studies investigated the association between migraine and myocardial infarction (1.12, 0.95 to 1.32) and five between migraine and death due to cardiovascular disease (1.03, 0.79 to 1.34).

Abstract: Only one study investigated the association between women who had migraine with aura and myocardial infarction and death due to cardiovascular disease, showing a twofold increased risk.

Abstract: CONCLUSION Migraine is associated with a twofold increased risk of ischaemic stroke, which is only apparent among people who have migraine with aura.

Abstract: Our results also suggest a higher risk among women and risk was further magnified for people with migraine who were aged less than 45, smokers, and women who used oral contraceptives.

Abstract: We did not find an overall association between any migraine and myocardial infarction or death due to cardiovascular disease.

Abstract: Too few studies are available to reliably evaluate the impact of modifying factors, such as migraine aura, on these associations.


Effect of Duration and Intermittency of Rifampin on Tuberculosis Treatment Outcomes: A Systematic Review and Meta-Analysis

Abstract: BACKGROUND Treatment regimens for active tuberculosis (TB) that are intermittent, or use rifampin during only the initial phase, offer practical advantages, but their efficacy has been questioned.

Abstract: We conducted a systematic review of treatment regimens for active TB, to assess the effect of duration and intermittency of rifampin use on TB treatment outcomes.

Abstract: METHODS AND FINDINGS PubMed, Embase, and the Cochrane CENTRAL database for clinical trials were searched for randomized controlled trials, published in English, French, or Spanish, between 1965 and June 2008.

Abstract: Selected studies utilized standardized treatment with rifampin-containing regimens.

Abstract: Studies reported bacteriologically confirmed failure and/or relapse in previously untreated patients with bacteriologically confirmed pulmonary TB.

Abstract: Pooled cumulative incidences of treatment outcomes and association with risk factors were computed with stratified random effects meta-analyses.

Abstract: Meta-regression was performed using a negative binomial regression model.

Abstract: A total of 57 trials with 312 arms and 21,472 participants were included in the analysis.

Abstract: Regimens utilizing rifampin only for the first 1-2 mo had significantly higher rates of failure, relapse, and acquired drug resistance, as compared to regimens that used rifampin for 6 mo.

Abstract: This was particularly evident when there was initial drug resistance to isoniazid, streptomycin, or both.

Abstract: On the other hand, there was little evidence of difference in failure or relapse with daily or intermittent schedules of treatment administration, although there was insufficient published evidence of the efficacy of twice-weekly rifampin administration throughout therapy.

Abstract: CONCLUSIONS TB treatment outcomes were significantly worse with shorter duration of rifampin, or with initial drug resistance to isoniazid and/or streptomycin.

Abstract: Treatment outcomes were similar with all intermittent schedules evaluated, but there is insufficient evidence to support administration of treatment twice weekly throughout therapy.


A novel set of DNA methylation markers in urine sediments for sensitive/specific detection of bladder cancer.

Abstract: PURPOSE This study aims to provide a better set of DNA methylation markers in urine sediments for sensitive and specific detection of bladder cancer.

Abstract: EXPERIMENTAL DESIGN Fifty-nine tumor-associated genes were profiled in three bladder cancer cell lines, a small cohort of cancer biopsies and urine sediments by methylation-specific PCR.

Abstract: Twenty-one candidate genes were then profiled in urine sediments from 132 bladder cancer patients (8 cases for stage 0a; 68 cases for stage I; 50 cases for stage II; 4 cases for stages III; and 2 cases for stage IV), 23 age-matched patients with noncancerous urinary lesions, 6 neurologic diseases, and 7 healthy volunteers.

Abstract: RESULTS Despite six incidences of four genes reported in 3 of 23 noncancerous urinary lesion patients analyzed, cancer-specific hypermethylation in urine sediments were reported for 15 genes (P < 0.05).

Abstract: Methylation assessment of an 11-gene set (SALL3, CFTR, ABCC6, HPR1, RASSF1A, MT1A, RUNX3, ITGA4, BCL2, ALX4, MYOD1, DRM, CDH13, BMP3B, CCNA1, RPRM, MINT1, and BRCA1) confirmed the existing diagnosis of 121 among 132 bladder cancer cases (sensitivity, 91.7%) with 87% accuracy.

Abstract: Significantly, more than 75% of stage 0a and 88% of stage I disease were detected, indicating its value in the early diagnosis of bladder cancer.

Abstract: Interestingly, the cluster of reported methylation markers used in the U.S. bladder cancers is distinctly different from that identified in this study, suggesting a possible epigenetic disparity between the American and Chinese cases.

Abstract: CONCLUSIONS Methylation profiling of an 11-gene set in urine sediments provides a sensitive and specific detection of bladder cancer.


Posttranslational Acetylation of α-Tubulin Constrains Protofilament Number in Native Microtubules

Abstract: BACKGROUND Microtubules are built from linear polymers of α-β tubulin dimers (protofilaments) that form a tubular quinary structure.

Abstract: Microtubules assembled from purified tubulin in vitro contain between 10 and 16 protofilaments; however, such structural polymorphisms are not found in cells.

Abstract: This discrepancy implies that factors other than tubulin constrain microtubule protofilament number, but the nature of these constraints is unknown.

Abstract: RESULTS Here, we show that acetylation of MEC-12 α-tubulin constrains protofilament number in C. elegans touch receptor neurons (TRNs).

Abstract: Whereas the sensory dendrite of wild-type TRNs is packed with a cross-linked bundle of long, 15-protofilament microtubules, mec-17;atat-2 mutants lacking α-tubulin acetyltransferase activity have short microtubules, rampant lattice defects, and variable protofilament number both between and within microtubules.

Abstract: All-atom molecular dynamics simulations suggest a model in which acetylation of lysine 40 promotes the formation of interprotofilament salt bridges, stabilizing lateral interactions between protofilaments and constraining quinary structure to produce stable, structurally uniform microtubules in vivo.

Abstract: CONCLUSIONS Acetylation of α-tubulin is an essential constraint on protofilament number in vivo.

Abstract: We propose a structural model in which this posttranslational modification promotes the formation of lateral salt bridges that fine-tune the association between adjacent protofilaments and enable the formation of uniform microtubule populations in vivo.


A KEY ROLE FOR OREXIN IN PANIC ANXIETY

Abstract: Panic disorder is a severe anxiety disorder with recurrent, debilitating panic attacks.

Abstract: In individuals with panic disorder there is evidence of decreased central gamma-aminobutyric acid (GABA) activity as well as marked increases in autonomic and respiratory responses after intravenous infusions of hypertonic sodium lactate.

Abstract: In a rat model of panic disorder, chronic inhibition of GABA synthesis in the dorsomedial-perifornical hypothalamus of rats produces anxiety-like states and a similar vulnerability to sodium lactate-induced cardioexcitatory responses.

Abstract: The dorsomedial-perifornical hypothalamus is enriched in neurons containing orexin (ORX, also known as hypocretin), which have a crucial role in arousal, vigilance and central autonomic mobilization, all of which are key components of panic.

Abstract: Here we show that activation of ORX-synthesizing neurons is necessary for developing a panic-prone state in the rat panic model, and either silencing of the hypothalamic gene encoding ORX (Hcrt) with RNAi or systemic ORX-1 receptor antagonists blocks the panic responses.

Abstract: Moreover, we show that human subjects with panic anxiety have elevated levels of ORX in the cerebrospinal fluid compared to subjects without panic anxiety.

Abstract: Taken together, our results suggest that the ORX system may be involved in the pathophysiology of panic anxiety and that ORX antagonists constitute a potential new treatment strategy for panic disorder.


Local Increases in Mechanical Tension Shape Compartment Boundaries by Biasing Cell Intercalations

Abstract: Mechanical forces play important roles during tissue organization in developing animals.

Abstract: Many tissues are organized into adjacent, nonmixing groups of cells termed compartments.

Abstract: Boundaries between compartments display a straight morphology and are associated with signaling centers that are important for tissue growth and patterning.

Abstract: Local increases in mechanical tension at cell junctions along compartment boundaries have recently been shown to prevent cell mixing and to maintain straight boundaries.

Abstract: The cellular mechanisms by which local increases in mechanical tension prevent cell mixing at compartment boundaries, however, remain poorly understood.

Abstract: Here, we have used live imaging and quantitative image analysis to determine cellular dynamics at and near the anteroposterior compartment boundaries of the Drosophila pupal abdominal epidermis.

Abstract: We show that cell mixing within compartments involves multiple cell intercalations.

Abstract: Frequency and orientation of cell intercalations are unchanged along the compartment boundaries; rather, an asymmetry in the shrinkage of junctions during intercalation is biased, resulting in cell rearrangements that suppress cell mixing.

Abstract: Simulations of tissue growth show that local increases in mechanical tension can account for this bias in junctional shrinkage.

Abstract: We conclude that local increases in mechanical tension maintain cell populations separate by influencing junctional rearrangements during cell intercalation.


MDA5 and PTPN2, two candidate genes for type 1 diabetes, modify pancreatic β-cell responses to the viral by-product double-stranded RNA

Abstract: beta-Cell destruction in type 1 diabetes (T1D) is at least in part consequence of a 'dialog' between beta-cells and immune system.

Abstract: This dialog may be affected by the individual's genetic background.

Abstract: We presently evaluated whether modulation of MDA5 and PTPN2, two candidate genes for T1D, affects beta-cell responses to double-stranded RNA (dsRNA), a by-product of viral replication.

Abstract: These genes were selected following comparison between known candidate genes for T1D and genes expressed in pancreatic beta-cells, as identified in previous array analysis.

Abstract: INS-1E cells and primary fluorescence-activated cell sorting-purified rat beta-cells were transfected with small interference RNAs (siRNAs) targeting MDA5 or PTPN2 and subsequently exposed to intracellular synthetic dsRNA (polyinosinic-polycitidilic acid-PIC).

Abstract: Real-time RT-PCR, western blot and viability assays were performed to characterize gene/protein expression and viability.

Abstract: PIC increased MDA5 and PTPN2 mRNA expression, which was inhibited by the specific siRNAs.

Abstract: PIC triggered apoptosis in INS-1E and primary beta-cells and this was augmented by PTPN2 knockdown (KD), although inhibition of MDA5 did not modify PIC-induced apoptosis.

Abstract: In contrast, MDA5 silencing decreased PIC-induced cytokine and chemokine expression, although inhibition of PTPN2 induced minor or no changes in these inflammatory mediators.

Abstract: These findings indicate that changes in MDA5 and PTPN2 expression modify beta-cell responses to dsRNA.

Abstract: MDA5 regulates inflammatory signals, whereas PTPN2 may function as a defence mechanism against pro-apoptotic signals generated by dsRNA.

Abstract: These two candidate genes for T1D may thus modulate beta-cell apoptosis and/or local release of inflammatory mediators in the course of a viral infection by acting, at least in part, at the pancreatic beta-cell level.


Tumor necrosis factor signaling mediates resistance to mycobacteria by inhibiting bacterial growth and macrophage death.

Abstract: Tumor necrosis factor (TNF), a key effector in controlling tuberculosis, is thought to exert protection by directing formation of granulomas, organized aggregates of macrophages and other immune cells.

Abstract: Loss of TNF signaling causes progression of tuberculosis in humans, and the increased mortality of Mycobacterium tuberculosis-infected mice is associated with disorganized necrotic granulomas, although the precise roles of TNF signaling preceding this endpoint remain undefined.

Abstract: We monitored transparent Mycobacterium marinum-infected zebrafish live to conduct a stepwise dissection of how TNF signaling operates in mycobacterial pathogenesis.

Abstract: We found that loss of TNF signaling caused increased mortality even when only innate immunity was operant.

Abstract: In the absence of TNF, intracellular bacterial growth and granuloma formation were accelerated and was followed by necrotic death of overladen macrophages and granuloma breakdown.

Abstract: Thus, TNF is not required for tuberculous granuloma formation, but maintains granuloma integrity indirectly by restricting mycobacterial growth within macrophages and preventing their necrosis.


Loss of negative regulation by Numb over Notch is relevant to human breast carcinogenesis

Abstract: The biological antagonism between Notch and Numb controls the proliferative/differentiative balance in development and homeostasis.

Abstract: Although altered Notch signaling has been linked to human diseases, including cancer, evidence for a substantial involvement of Notch in human tumors has remained elusive.

Abstract: Here, we show that Numb-mediated control on Notch signaling is lost in ∼50% of human mammary carcinomas, due to specific Numb ubiquitination and proteasomal degradation.

Abstract: Mechanistically, Numb operates as an oncosuppressor, as its ectopic expression in Numb-negative, but not in Numb-positive, tumor cells inhibits proliferation.

Abstract: Increased Notch signaling is observed in Numb-negative tumors, but reverts to basal levels after enforced expression of Numb.

Abstract: Conversely, Numb silencing increases Notch signaling in normal breast cells and in Numb-positive breast tumors.

Abstract: Finally, growth suppression of Numb-negative, but not Numb-positive, breast tumors can be achieved by pharmacological inhibition of Notch.

Abstract: Thus, the Numb/Notch biological antagonism is relevant to the homeostasis of the normal mammary parenchyma and its subversion contributes to human mammary carcinogenesis.


Cortical Dynein Controls Microtubule Dynamics to Generate Pulling Forces that Position Microtubule Asters

Abstract: Dynein at the cortex contributes to microtubule-based positioning processes such as spindle positioning during embryonic cell division and centrosome positioning during fibroblast migration.

Abstract: To investigate how cortical dynein interacts with microtubule ends to generate force and how this functional association impacts positioning, we have reconstituted the 'cortical' interaction between dynein and dynamic microtubule ends in an in vitro system using microfabricated barriers.

Abstract: We show that barrier-attached dynein captures microtubule ends, inhibits growth, and triggers microtubule catastrophes, thereby controlling microtubule length.

Abstract: The subsequent interaction with shrinking microtubule ends generates pulling forces up to several pN. By combining experiments in microchambers with a theoretical description of aster mechanics, we show that dynein-mediated pulling forces lead to the reliable centering of microtubule asters in simple confining geometries.

Abstract: Our results demonstrate the intrinsic ability of cortical microtubule-dynein interactions to regulate microtubule dynamics and drive positioning processes in living cells.


Breast cancer risk factors in relation to breast density (United States)

Abstract: Evaluate known breast cancer risk factors in relation to breast density.

Abstract: We examined factors in relation to breast density in 144,018 New Hampshire (NH) women with at least one mammogram recorded in a statewide mammography registry.

Abstract: Mammographic breast density was measured by radiologists using the BI-RADS classification; risk factors of interest were obtained from patient intake forms and questionnaires.

Abstract: Initial analyses showed a strong inverse influence of age and body mass index (BMI) on breast density.

Abstract: In addition, women with late age at menarche, late age at first birth, premenopausal women, and those currently using hormone therapy (HT) tended to have higher breast density, while those with greater parity tended to have less dense breasts.

Abstract: Analyses stratified on age and BMI suggested interactions, which were formally assessed in a multivariable model.

Abstract: The impact of current HT use, relative to nonuse, differed across age groups, with an inverse association in younger women, and a positive association in older women (p < 0.0001 for the interaction).

Abstract: The positive effects of age at menarche and age at first birth, and the inverse influence of parity were less apparent in women with low BMI than in those with high BMI (p = 0.04, p < 0.0001 and p = 0.01, respectively, for the interactions).

Abstract: We also noted stronger positive effects for age at first birth in postmenopausal women (p = 0.004 for the interaction).

Abstract: The multivariable model indicated a slight positive influence of family history of breast cancer.

Abstract: The influence of age at menarche and reproductive factors on breast density is less evident in women with high BMI.

Abstract: Density is reduced in young women using HT, but increased in HT users of age 50 or more.


Perspective: repression of competition and the evolution of cooperation.

Abstract: Repression of competition within groups joins kin selection as the second major force in the history of life shaping the evolution of cooperation.

Abstract: When opportunities for competition against neighbors are limited within groups, individuals can increase their own success only by enhancing the efficiency and productivity of their group.

Abstract: Thus, characters that repress competition within groups promote cooperation and enhance group success.

Abstract: Leigh first expressed this idea in the context of fair meiosis, in which each chromosome has an equal chance of transmission via gametes.

Abstract: Randomized success means that each part of the genome can increase its own success only by enhancing the total number of progeny and thus increasing the success of the group.

Abstract: Alexander used this insight about repression of competition in fair meiosis to develop his theories for the evolution of human sociality.

Abstract: Alexander argued that human social structures spread when they repress competition within groups and promote successful group-against-group competition.

Abstract: Buss introduced a new example with his suggestion that metazoan success depended on repression of competition between cellular lineages.

Abstract: Maynard Smith synthesized different lines of thought on repression of competition.

Abstract: In this paper, I develop simple mathematical models to illustrate the main processes by which repression of competition evolves.

Abstract: With the concepts made clear, I then explain the history of the idea.

Abstract: I finish by summarizing many new developments in this subject and the most promising lines for future study.


Article

Abstract: IBD is characterized by uncontrolled immune responses in inflamed mucosa, with dominance of IL-17-producing cells and deficiency of Treg cells.

Abstract: The aim of this study was to explore the effect and mechanisms of RA, the ligand of RARalpha, on immune responses in human and murine colitis.

Abstract: Colonic biopsies from patients with UC were cultured and treated with RA as the agonist of RARalpha or LE135 as the antagonist of RARalpha.

Abstract: Expressions of IL-17 and FOXP3 were detected by immunohistochemistry.

Abstract: Murine colitis was induced by intrarectal administration with TNBS at Day 1.

Abstract: Mice were then i.p.-treated with RA or LE135 daily for 7 days.

Abstract: Cytokine levels in the cultures of mouse LPMCs were measured.

Abstract: Expressions of FOXP3 and IL-17 in colon tissues or MLN were detected by immunohistological analysis.

Abstract: Body weight and colon inflammation were evaluated.

Abstract: RA treatment up-regulated FOXP3 expression and down-regulated IL-17 expression in colon biopsies of patients and in colon tissues and MLN of mice with colitis compared with controls.

Abstract: LPMCs from RA-treated mice produced lower levels of proinflammatory cytokines (TNF-alpha, IL-1beta, IL-17) but more regulatory cytokines (IL-10, TGF-beta) compared with that of untreated mice.

Abstract: LE135 showed the opposite effect of RA.

Abstract: Furthermore, RA ameliorated TNBS-induced colitis in a dose-dependent manner, as seen by improved body weight and colon inflammation.

Abstract: RA down-regulates colon inflammatory responses in patients with IBD in vitro and in murine colitis in vivo, representing a potential therapeutic approach in IBD treatment.


Multiple RNA surveillance pathways limit aberrant expression of iron uptake mRNAs and prevent iron toxicity in S. cerevisiae.

Abstract: Tight regulation of the expression of mRNAs encoding iron uptake proteins is essential to control iron homeostasis and avoid intracellular iron toxicity.

Abstract: We show that many mRNAs encoding iron uptake or iron mobilization proteins are expressed in iron-replete conditions in the absence of the S. cerevisiae RNase III ortholog Rnt1p or of the nuclear exosome component Rrp6p.

Abstract: Extended forms of these mRNAs accumulate in the absence of Rnt1p or of the 5'-->3' exonucleases Xrn1p and Rat1p, showing that multiple degradative pathways contribute to the surveillance of aberrant forms of these transcripts.

Abstract: RNase III-deficient cells are hypersensitive to high iron concentrations, suggesting that Rnt1p-mediated RNA surveillance is required to prevent iron toxicity.

Abstract: These results show that RNA surveillance through multiple ribonucleolytic pathways plays a role in iron homeostasis in yeast to avoid the potentially toxic effects of the expression of the iron starvation response in iron-replete conditions.


Differential regulation of adherens junction dynamics during apical-basal polarization.

Abstract: Adherens junctions (AJs) in epithelial cells are constantly turning over to modulate adhesion properties under various physiological and developmental contexts, but how such AJ dynamics are regulated during the apical-basal polarization of primary epithelia remains unclear.

Abstract: Here, we used new and genetically validated GFP markers of Drosophila E-cadherin (DE-cadherin, hereafter referred to as DE-Cad) and β-catenin (Armadillo, Arm) to quantitatively assay the in vivo dynamics of biosynthetic turnover and membrane redistribution by fluorescence recovery after photobleaching (FRAP) assays.

Abstract: Our data showed that membrane DE-Cad and Arm in AJs of polarizing epithelial cells had much faster biosynthetic turnover than in polarized cells.

Abstract: Fast biosynthetic turnover of membrane DE-Cad is independent of actin- and dynamin-based trafficking, but is microtubule-dependent.

Abstract: Furthermore, Arm in AJs of polarizing cells showed a faster and diffusion-based membrane redistribution that was both quantitatively and qualitatively different from the slower and exchange-based DE-Cad membrane distribution, indicating that the association of Arm with DE-Cad is more dynamic in polarizing cells, and only becomes stable in polarized epithelial cells.

Abstract: Consistently, biochemical assays showed that the binding of Arm to DE-Cad is weaker in polarizing cells than in polarized cells.

Abstract: Our data revealed that the molecular interaction between DE-Cad and Arm is modulated during apical-basal polarization, suggesting a new mechanism that might be crucial for establishing apical-basal polarity through regulating the AJ dynamics.


Phylogenetic analysis of mRNA polyadenylation sites reveals a role of transposable elements in evolution of the 3′-end of genes

Abstract: mRNA polyadenylation is an essential step for the maturation of almost all eukaryotic mRNAs, and is tightly coupled with termination of transcription in defining the 3'-end of genes.

Abstract: Large numbers of human and mouse genes harbor alternative polyadenylation sites [poly(A) sites] that lead to mRNA variants containing different 3'-untranslated regions (UTRs) and/or encoding distinct protein sequences.

Abstract: Here, we examined the conservation and divergence of different types of alternative poly(A) sites across human, mouse, rat and chicken.

Abstract: We found that the 3'-most poly(A) sites tend to be more conserved than upstream ones, whereas poly(A) sites located upstream of the 3'-most exon, also termed intronic poly(A) sites, tend to be much less conserved.

Abstract: Genes with longer evolutionary history are more likely to have alternative polyadenylation, suggesting gain of poly(A) sites through evolution.

Abstract: We also found that nonconserved poly(A) sites are associated with transposable elements (TEs) to a much greater extent than conserved ones, albeit less frequently utilized.

Abstract: Different classes of TEs have different characteristics in their association with poly(A) sites via exaptation of TE sequences into polyadenylation elements.

Abstract: Our results establish a conservation pattern for alternative poly(A) sites in several vertebrate species, and indicate that the 3'-end of genes can be dynamically modified by TEs through evolution.


Inter- and Intra-Host Viral Diversity in a Large Seasonal DENV2 Outbreak

Abstract: BACKGROUND High genetic diversity at both inter- and intra-host level are hallmarks of RNA viruses due to the error-prone nature of their genome replication.

Abstract: Several groups have evaluated the extent of viral variability using different RNA virus deep sequencing methods.

Abstract: Although much of this effort has been dedicated to pathogens that cause chronic infections in humans, few studies investigated arthropod-borne, acute viral infections.

Abstract: METHODS AND PRINCIPAL FINDINGS We deep sequenced the complete genome of ten DENV2 isolates from representative classical and severe cases sampled in a large outbreak in Brazil using two different approaches.

Abstract: Analysis of the consensus genomes confirmed the larger extent of the 2010 epidemic in comparison to a previous epidemic caused by the same viruses in another city two years before (genetic distance = 0.002 and 0.0008 respectively).

Abstract: Analysis of viral populations within the host revealed a high level of conservation.

Abstract: After excluding homopolymer regions of 454/Roche generated sequences, we found 10 to 44 variable sites per genome population at a frequency of >1%, resulting in very low intra-host genetic diversity.

Abstract: While up to 60% of all variable sites at intra-host level were non-synonymous changes, only 10% of inter-host variability resulted from non-synonymous mutations, indicative of purifying selection at the population level.

Abstract: CONCLUSIONS AND SIGNIFICANCE Despite the error-prone nature of RNA-dependent RNA-polymerase, dengue viruses maintain low levels of intra-host variability.


Serum Soluble Corin Deficiency Predicts Major Disability within 3 Months after Acute Stroke

Abstract: OBJECTIVE Serum soluble corin has been associated with stroke.

Abstract: However, whether it is associated with stroke prognosis has not yet been studied.

Abstract: Therefore, we aimed to study the association of serum soluble corin with risk of poor outcomes within 3 months after stroke.

Abstract: METHODS We followed 522 stroke patients for 3 months to identify major disability, death and vascular events.

Abstract: Serum soluble corin was measured at baseline for all participants.

Abstract: Logistic regression was used to examine the associations of baseline serum soluble corin with outcomes of stroke, adjusting for age, sex, baseline NIHSS score, hours from onset to hospitalization, smoking, drinking, hypertension, diabetes, coronary heart disease, atrial fibrillation, family history of stroke, and stroke subtype.

Abstract: RESULTS Patients with high corin had a significantly lower crude risk for the composite outcome of major disability or death (OR = 0.64, 95%CI: 0.43-0.96) than patients with low corin (the lowest tertile).

Abstract: After adjustment for age and baseline NIHSS score, patients with high corin still had a significantly lower risk for the composite outcome of major disability or death (OR = 0.60, 95%CI: 0.36-0.99).

Abstract: This association became bottom line significant after additionally adjusting for other conventional factors (OR = 0.61, P = 0.058).

Abstract: No association was found between serum soluble corin and other composite outcomes.

Abstract: CONCLUSION Serum soluble corin deficiency predicted risk for major disability within 3 months after stroke, independent of baseline neurological deficient.

Abstract: Our results may indicate a probable role of corin in stroke prognosis.


Altered PTEN expression; a diagnostic marker for differentiating normal, hyperplastic and neoplastic endometrium

Abstract: BACKGROUND Different molecular alterations have been described in endometrioid endometrial carcinoma (EECA).

Abstract: Among them the most frequently altered is loss of the PTEN protein, a tumor suppressor gene.

Abstract: The purpose of this study was to evaluate the expression pattern of PTEN gene in normal, hyperplastic and neoplastic endometrium.

Abstract: METHODS In a study in a referral gynecologic hospital in Tehran, Iran, immunohistochemical (IHC) evaluation of PTEN was performed on 87 consecutive specimens to the following three groups; group A- normal proliferative endometrium(n = 29); group B- hyperplastic endometrium [including simple hyperplasia without atypia(n = 21) and complex hyperplasia with atypia (n = 8)] and group C- EECA(n = 29).

Abstract: Immunostaining of cells was analyzed by arbitrary quantitative methods according to both slide's area staining and intensity of color reaction.

Abstract: RESULTS PTEN immunoreactivity was present in all normal proliferative endometrium, all simple hyperplasia, 75% of atypical complex hyperplasia and in 48% of EECA (P < 0.001).

Abstract: The intensity of PTEN reaction was significantly higher in group with proliferative endometrium than hyperplastic endometrium and EECA (P < 0.001).

Abstract: CONCLUSION PTEN expression was significantly higher in cyclical endometrium than in atypical hyperplasia and endometrioid carcinoma.


Care Seeking for Neonatal Illness in Low- and Middle-Income Countries: A Systematic Review

Abstract: BACKGROUND Despite recent achievements to reduce child mortality, neonatal deaths continue to remain high, accounting for 41% of all deaths in children under five years of age worldwide, of which over 90% occur in low- and middle-income countries (LMICs).

Abstract: Infections are a leading cause of death and limitations in care seeking for ill neonates contribute to high mortality rates.

Abstract: As estimates for care-seeking behaviors in LMICs have not been studied, this review describes care seeking for neonatal illnesses in LMICs, with particular attention to type of care sought.

Abstract: METHODS AND FINDINGS We conducted a systematic literature review of studies that reported the proportion of caregivers that sought care for ill or suspected ill neonates in LMICs.

Abstract: The initial search yielded 784 studies, of which 22 studies described relevant data from community household surveys, facility-based surveys, and intervention trials.

Abstract: The majority of studies were from South Asia (n = 17/22), set in rural areas (n = 17/22), and published within the last 4 years (n = 18/22).

Abstract: Of the 9,098 neonates who were ill or suspected to be ill, 4,320 caregivers sought some type of care, including care from a health facility (n = 370) or provider (n = 1,813).

Abstract: Care seeking ranged between 10% and 100% among caregivers with a median of 59%.

Abstract: Care seeking from a health care provider yielded a similar range and median, while care seeking at a health care facility ranged between 1% and 100%, with a median of 20%.

Abstract: Care-seeking estimates were limited by the few studies conducted in urban settings and regions other than South Asia.

Abstract: There was a lack of consistency regarding illness, care-seeking, and care provider definitions.

Abstract: CONCLUSIONS There is a paucity of data regarding newborn care-seeking behaviors; in South Asia, care seeking is low for newborn illness, especially in terms of care sought from health care facilities and medically trained providers.

Abstract: There is a need for representative data to describe care-seeking patterns in different geographic regions and better understand mechanisms to enhance care seeking during this vulnerable time period.


Synaptic Incorporation of AMPA Receptors during LTP Is Controlled by a PKC Phosphorylation Site on GluR1

Abstract: Incorporation of GluR1-containing AMPA receptors into synapses is essential to several forms of neural plasticity, including long-term potentiation (LTP).

Abstract: Numerous signaling pathways that trigger this process have been identified, but the direct modifications of GluR1 that control its incorporation into synapses are unclear.

Abstract: Here, we show that phosphorylation of GluR1 by PKC at a highly conserved serine 818 residue is increased during LTP and critical for LTP expression.

Abstract: GluR1 is phosphorylated by PKC at this site in vitro and in vivo.

Abstract: In addition, acute phosphorylation at GluR1 S818 by PKC, as well as a phosphomimetic mutation, promotes GluR1 synaptic incorporation.

Abstract: Conversely, preventing GluR1 S818 phosphorylation reduces LTP and blocks PKC-driven synaptic incorporation of GluR1.

Abstract: We conclude that the phosphorylation of GluR1 S818 by PKC is a critical event in the plasticity-driven synaptic incorporation of AMPA receptors.