Genome-wide survey of remote homologues for protein domain superfamilies of known structure reveals unequal distribution across structural classes.
Domains are the basic building blocks of proteins which can combine to give rise to different domain architectures. Annotation of domains in a sequence is the first step towards understanding the biological function. Since there are a limited number of folds and evolutionarily related proteins have a similar structure, function can be inferred through remote homology. Computational sequence searches were performed for remote homologues on genomes of around ∼160 000 different organisms, starting from nearly 11 000 superfamily queries of known structure. Case studies revealed that most of the associated domains are involved in the same biological process. Using all the proteins predicted to have at least one structural domain, a coverage of 61% of Pfam families was achieved which is higher than the existing methods (43.36% by SIFTS). Taxonomic analysis of the proteins revealed 493 superfamilies in all the major kingdoms of life and a few lateral gene transfers between viruses and cellular organisms. The distribution of remote homologues across different classes, folds and superfamilies was studied and reveals that sequences are unequally distributed across structural classes. Finally, domain architectures were computed for the homologues and these data were compiled for each superfamily and organism.


Pavlov's dog associative learning demonstrated on synaptic-like organic transistors.
In this letter, we present an original demonstration of an associative learning neural network inspired by the famous Pavlov's dogs experiment. A single nanoparticle organic memory field effect transistor (NOMFET) is used to implement each synapse. We show how the physical properties of this dynamic memristive device can be used to perform low-power write operations for the learning and implement short-term association using temporal coding and spike-timing-dependent plasticity-based learning. An electronic circuit was built to validate the proposed learning scheme with packaged devices, with good reproducibility despite the complex synaptic-like dynamic of the NOMFET in pulse regime.


RACK1 regulates directional cell migration by acting on G betagamma at the interface with its effectors PLC beta and PI3K gamma.
Migration of cells up the chemoattractant gradients is mediated by the binding of chemoattractants to G protein-coupled receptors and activation of a network of coordinated excitatory and inhibitory signals. Although the excitatory process has been well studied, the molecular nature of the inhibitory signals remains largely elusive. Here we report that the receptor for activated C kinase 1 (RACK1), a novel binding protein of heterotrimeric G protein betagamma (G betagamma) subunits, acts as a negative regulator of directed cell migration. After chemoattractant-induced polarization of Jurkat and neutrophil-like differentiated HL60 (dHL60) cells, RACK1 interacts with G betagamma and is recruited to the leading edge. Down-regulation of RACK1 dramatically enhances chemotaxis of cells, whereas overexpression of RACK1 or a fragment of RACK1 that retains G betagamma-binding capacity inhibits cell migration. Further studies reveal that RACK1 does not modulate cell migration through binding to other known interacting proteins such as PKC beta and Src. Rather, RACK1 selectively inhibits G betagamma-stimulated phosphatidylinositol 3-kinase gamma (PI3K gamma) and phospholipase C (PLC) beta activity, due to the competitive binding of RACK1, PI3K gamma, and PLC beta to G betagamma. Taken together, these findings provide a novel mechanism of regulating cell migration, i.e., RACK1-mediated interference with G betagamma-dependent activation of key effectors critical for chemotaxis.


Feasibility study of a plastic scintillating plate-based treatment beam fluence monitoring system for use in pencil beam scanning proton therapy.
The purpose of this study was to describe a plastic scintillating plate-based gantry-attachable dosimetry system for pencil beam scanning proton therapy to monitor entrance proton beam fluence, and to evaluate the dosimetric characteristics of this system and its feasibility for clinical use. The dosimetry system, consisting of a plastic scintillating plate and a CMOS camera, was attached to a dedicated scanning nozzle and scintillation during proton beam irradiation was recorded. Dose distribution was calculated from the accumulated recorded frames. The dosimetric characteristics (energy dependency, dose linearity, dose rate dependency, and reproducibility) of the gantry-attachable dosimetry system for use with therapeutic proton beams were measured, and the feasibility of this system during clinical use was evaluated by determining selected quality assurance items at our institution. The scintillating plate shortened the range of the proton beam by the water-equivalent thickness of the plate and broadened the spatial profile of the single proton spot by 11% at 70 MeV. The developed system functioned independently of the beam energy (<1.3%) and showed dose linearity, and also functioned independently of the dose rate. The feasibility of the system for clinical use was evaluated by comparing the measured quality assurance dose distribution to that of the treatment planning system. The gamma passing rate with a criterion of 3%/3 mm was 97.58%. This study evaluated the dosimetric characteristics of a plastic scintillating plate-based dosimetry system for use with scanning proton beams. The ability to account for the interference of the dosimetry system on the therapeutic beam enabled offline monitoring of the entrance beam fluence of the pencil beam scanning proton therapy independent of the treatment system with high resolution and in a cost-effective manner.


Processing effect on soybean storage proteins and their relationship with tofu quality.
Tofu was prepared from 13 soybean varieties according to three different methods (bench, pilot, and production methods). Different soybean varieties showed significant differences in storage protein composition (glycinin and beta-conglycinin). The beta-conglycinin (7S) and glycinin (11S) contents were 7.3-9.9 and 14.1-22.9% on the dry matter basis, respectively. The 11S/7S protein ratio varied from 1.64 to 2.51 among the varieties. Glycinin content and 11S/7S protein ratio of the 13 varieties did not change significantly from soy milk to tofu for the production and pilot methods. Soybean 11S/7S protein ratio positively correlated with the 11S/7S ratio of soy milk and tofu (0.57 < or = r < or = 0.83, p < or = 0.01). The correlation coefficient depended on the processing method. Processing method affected 7S and 11S protein contents of tofu and their contribution to tofu hardness, yield, and sensory quality. This may explain in part the contradictory findings of the relationships between storage proteins and tofu quality because processing methods differed in various studies.


Seeing white: Qualia in the context of decoding population codes.
When the nervous system is presented with multiple simultaneous inputs of some variable, such as wavelength or disparity, they can be combined to give rise to qualitatively new percepts that cannot be produced by any single input value. For example, there is no single wavelength that appears white. Many models of decoding neural population codes have problems handling multiple inputs, either attempting to extract a single value of the input parameter or, in some cases, registering the presence of multiple inputs without synthesizing them into something new. These examples raise a more general issue regarding the interpretation of population codes. We propose that population decoding involves not the extraction of specific values of the physical inputs, but rather a transformation from the input space to some abstract representational space that is not simply related to physical parameters. As a specific example, a four-layer network is presented that implements a transformation from wavelength to a high-level hue-saturation color space.


Differential expression of a putative CarD-like transcriptional regulator, LtpA, in Borrelia burgdorferi.
The availability of microbial genome information has provided a fruitful opportunity for studying regulatory networks in a variety of pathogenic bacteria. In an initial effort to elucidate regulatory networks potentially involved in differential gene expression by the Lyme disease pathogen Borrelia burgdorferi, we have been investigating the functions and regulation of putative transcriptional regulatory factors predicted to be encoded within the B. burgdorferi genome. Herein we report the regulation of one of the predicted transcriptional regulators, LtpA (BB0355), which is homologous to the transcriptional regulator CarD from Myxococcus xanthus. LtpA expression was assessed in response to various environmental stimuli. Immunoblot and quantitative reverse transcription-PCR analyses revealed that unlike many well-characterized differentially regulated Borrelia genes whose expression is induced by elevated temperature, the expression of LtpA was significantly downregulated when spirochetes were grown at an elevated temperature (37 degrees C), as well as when the bacteria were cultivated in a mammalian host-adapted environment. In contrast, LtpA was induced at a lower culture temperature (23 degrees C). Further analyses indicated that the downregulation of LtpA was not dependent on the Rrp2-RpoN-RpoS regulatory pathway, which is involved in the downregulation of OspA when B. burgdorferi is grown in a mammalian host-adapted environment. LtpA protein levels in B. burgdorferi were unaltered in response to changes in the pH in the borrelial cultures. Multiple attempts to generate an LtpA-deficient mutant were unsuccessful, which has hampered the elucidation of its role in pathogenesis. Given that LtpA is exclusively expressed during borrelial cultivation at a lower temperature, a parameter that has been widely used as a surrogate condition to mimic B. burgdorferi in unfed (flat) ticks, and because LtpA is homologous to a known transcriptional regulator, we postulate that LtpA functions as a regulator modulating the expression of genes important to B. burgdorferi's survival within its arthropod vector.


What Do Parents Think About School Meals? An Exploratory Study of Rural Middle School Parents' Perceptions.
Interventions targeting school meals have been used to combat obesity in rural youth. Parents play a powerful role in childhood nutrition; however, we know little about parents' perceptions of school meal programs. This study aimed to understand parents' perceptions of school meal programs. Surveys were administered to middle school parents (n = 576) at six schools in a rural state. We conducted univariate and multivariate analyses to measure parents' overall perceptions of the school meals program and to identify differences in perceptions by their children's participation in the Free and Reduced Price School Meals (F&RM) program. Parents had largely negative views of school meals but agreed that they met students' needs. Results of the multivariate analysis revealed differences in perceptions by participation in F&RM including the relative importance of meal components. Including parents in meal-related experiences and decision making could improve their perceptions and increase children's participation in school lunch programs.


Cellular metabolism of 3'-azido-2',3'-dideoxyuridine with formation of 5'-O-diphosphohexose derivatives by previously unrecognized metabolic pathways for 2'-deoxyuridine analogs.
3'-Azido-2',3'-dideoxyuridine (AzdU, CS-87) is a potent inhibitor of human immunodeficiency virus replication in human peripheral blood mononuclear cells (PBMC) with limited toxicity for human bone marrow cells (BMC). In the present study, metabolism of AzdU was investigated in human PBMC and BMC after exposure of cells to 2 or 10 microM [3H]AzdU. 3'-Azido-2',3'-dideoxyuridine-5'-monophosphate (AzdU-MP) was the predominant metabolite, representing approximately 55 to 65% of intracellular radioactivity in both PBMC and BMC at all times. The AzdU-5'-diphosphate and -5'-triphosphate intracellular levels were 10- to 100-fold lower than the AzdU-MP levels and, of note, AzdU-5'-triphosphate was not detected in human BMC. Using anion exchange chromatography, a new peak of radioactivity, distinct from any known anabolites, was detected. This chromatographic peak was found to be resistant to alkaline phosphatase but was hydrolyzed by 5'-phosphodiesterase, yielding AzdU-MP. Incubation of [3H]AzdU and D-[1-14C]glucose in PBMC and BMC produced a double-labeled peak with the same retention time as the anabolite, suggesting formation of a hexose derivative of AzdU. A novel high performance liquid chromatography method was developed that allowed for the separation of nucleosides, nucleotides, and carbohydrate derivatives thereof. Using this highly specific method, the putative AzdU-hexose actually was separated into two chromatographic peaks. These novel metabolites were identified as 3'-azido-2',3'-dideoxyuridine-5'-O-diphosphoglucose and 3'-azido-2',3'-dideoxyuridine-5'-O-diphospho-N-acetylglucosamine. Following 48 hr of incubation with [3H] AzdU, as much as 20 and 30% of these AzdU metabolites accumulated in PBMC and BMC, respectively. When AzdU was removed from the cell cultures, intracellular AzdU diphosphohexose concentrations decayed in a monophasic manner, with an elimination half-life of 14.3 hr. By 48 hr, levels of 0.3 pmol/10(6) cells were still detected, reflecting a gradual anabolism of these metabolites. Elimination of AzdU-MP and AzdU-5'-diphosphate was characterized by a two-phase process, with a short initial half-life of 0.83 and 0.24 hr and a long terminal half-life of 14.10 and 8.24 hr, respectively. Similar diphosphohexoses of deoxyuridine (dUrd) were also detected in human PBMC and BMC after exposure to [3H]dUrd, suggesting that dUrd derivatives are metabolized in a similar manner. In summary, the discovery of novel metabolic pathways for dUrd analogs demonstrates that AzdU has unique metabolic features that may contribute to the low toxicity of this anti-HIV agent in human BMC and also affect its mechanism of action.(ABSTRACT TRUNCATED AT 400 WORDS)


Raman micro-spectroscopy for rapid screening of oral squamous cell carcinoma.
Raman spectroscopy can provide a molecular-level fingerprint of the biochemical composition and structure of cells with excellent spatial resolution and could be useful to monitor changes in composition for dysplasia and early, non-invasive cancer diagnosis (carcinoma in situ), both ex-vivo and in vivo. In this study, we demonstrate this potential by collecting Raman spectra of the nucleoli, nuclei and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (pre-cancerous, DOK) cell lines and from normal oral epithelial primary cell cultures, in vitro, which were then analysed by principal component analysis (PCA) as a multivariate statistical method to discriminate the spectra. Results show significant discrimination between cancer and normal cell lines. Furthermore, the dysplastic and cancer cell lines could be discriminated based on the spectral profiles of the cytoplasmic regions. The principal component loading plot, which elucidates the biochemical features responsible for the discrimination, showed significant contributions of nucleic acid and proteins for nucleolar and nuclear sites and variation in features of lipids for the cytoplasmic area. This technique may provide a rapid screening method and have potential use in the diagnosis of dysplasia and early, non-invasive oral cancer, the treatment of which involves much less extensive and complex surgery and a reduction in associated co-morbidity for the patient.


Gene expression profile of peripheral blood lymphocytes from renal cell carcinoma patients treated with IL-2, interferon-α and dendritic cell vaccine.
Lymphocytes are a key component of the immune system and their differentiation and function are directly influenced by cancer. We examined peripheral blood lymphocyte (PBL) gene expression as a biomarker of illness and treatment effect using the Affymetrix Human Gene ST1 platform in patients with metastatic renal cell carcinoma (mRCC) who received combined treatment with IL-2, interferon-?-2a and dendritic cell vaccine. We examined gene expression, cytokine levels in patient serum and lymphocyte subsets as determined by flow cytometry (FCM). Pre-treatment PBLs from patients with mRCC exhibit a gene expression profile and serum cytokine profile consistent with inflammation and proliferation not found in healthy donors (HD). PBL gene expression from patients with mRCC showed increased mRNA of genes involved with T-cell and T(REG)-cell activation pathways, which was also reflected in lymphocyte subset distribution. Overall, PBL gene expression post-treatment (POST) was not significantly different than pre-treatment (PRE). Nevertheless, treatment related changes in gene expression (post-treatment minus pre-treatment) revealed an increased expression of T-cell and B-cell receptor signaling pathways in responding (R) patients compared to non-responding (NR) patients. In addition, we observed down-regulation of T(REG)-cell pathways post-treatment in R vs. NR patients. While exploratory in nature, this study supports the hypothesis that enhanced inflammatory cytotoxic pathways coupled with blunting of the regulatory pathways is necessary for effective anti-cancer activity associated with immune therapy. This type of analysis can potentially identify additional immune therapeutic targets in patients with mRCC.


Biological and biochemical activities of Vipera berus (European viper) venom.
Vipera berus is widely distributed throughout the northern part of Europe and Asia. Characterization of several toxic effects of its venom in the mouse, as well as of in vitro enzymatic activities was performed. Vipera berus venom displayed in vitro proteolytic, fibrinolytic, anticoagulant, and phospholipase A2 activities. The i.p. LD50 of the venom for Swiss mice was 0.86 micrograms/g (95% confidence limits 0.71-1.01 microgram/g). Significant local tissue-damaging effects, including edema, hemorrhage and myonecrosis, were observed. The local edema was characterized by rapid onset, reaching a maximum after 0.5-1 hr, and with dose-dependent persistence. The hemorrhagic potency was measured by a skin test, giving a minimum hemorrhagic dose value of 3.2 micrograms. The venom also induced a moderate local myonecrosis, evidenced by histological evaluation of injected tissue (gastrocnemius), and by biochemical parameters (increase of plasma creatine kinase activity, and decrease of muscle residual MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide)-reducing activity). Characterization of the venom by SDS-polyacrylamide gel electrophoresis revealed 10 (reduced) or 11 (unreduced) main protein bands, which were further analyzed in relation to mol. wt and relative concentration by densitometry. A rabbit antiserum to V. berus venom recognized all main venom bands by immunoblotting. This antiserum cross-reacted to a variable extent with several crotaline venoms, as assessed by enzyme immunoassay.


Prototype classification: insights from machine learning.
We shed light on the discrimination between patterns belonging to two different classes by casting this decoding problem into a generalized prototype framework. The discrimination process is then separated into two stages: a projection stage that reduces the dimensionality of the data by projecting it on a line and a threshold stage where the distributions of the projected patterns of both classes are separated. For this, we extend the popular mean-of-class prototype classification using algorithms from machine learning that satisfy a set of invariance properties. We report a simple yet general approach to express different types of linear classification algorithms in an identical and easy-to-visualize formal framework using generalized prototypes where these prototypes are used to express the normal vector and offset of the hyperplane. We investigate non-margin classifiers such as the classical prototype classifier, the Fisher classifier, and the relevance vector machine. We then study hard and soft margin classifiers such as the support vector machine and a boosted version of the prototype classifier. Subsequently, we relate mean-of-class prototype classification to other classification algorithms by showing that the prototype classifier is a limit of any soft margin classifier and that boosting a prototype classifier yields the support vector machine. While giving novel insights into classification per se by presenting a common and unified formalism, our generalized prototype framework also provides an efficient visualization and a principled comparison of machine learning classification.


The structural basis of TGF-beta, bone morphogenetic protein, and activin ligand binding.
The transforming growth factor-beta (TGF-beta) superfamily is a large group of structurally related growth factors that play prominent roles in a variety of cellular processes. The importance and prevalence of TGF-beta signaling are also reflected by the complex network of check points that exist along the signaling pathway, including a number of extracellular antagonists and membrane-level signaling modulators. Recently, a number of important TGF-beta crystal structures have emerged and given us an unprecedented clarity on several aspects of the signal transduction process. This review will highlight these latest advances and present our current understanding on the mechanisms of specificity and regulation on TGF-beta signaling outside the cell.


Notch 1 signaling is active in ovarian cancer.
OBJECTIVE.: Despite advances in chemotherapy and radical surgery, most advanced stage ovarian cancer patients die from their disease, highlighting the need for the development of novel treatment strategies. The Notch signaling pathway plays an important role in cellular differentiation, proliferation and apoptosis. We hypothesized that the active form of Notch 1, the Notch 1 intracellular domain (NICD), would be overexpressed in ovarian cancer cells and that depletion of NICD would lead to growth reduction. METHODS.: Following institutional review board approval, NICD expression was analyzed in human ovarian cancer specimens as well as the ovarian cancer cell lines OVCAR3, SKOV3, and CaOV3. In addition, the effects of Notch 1 depletion on ovarian cancer cell growth were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) growth assay for 6 days following transfection with siRNA against Notch 1. RESULTS.: Western blot analysis revealed abundant NICD expression in all 3 ovarian cancer cell lines, as well as in 16 of 21 (76%) human ovarian cancer samples. Following treatment with Notch 1 siRNA, expression of NICD was greatly reduced in all three cell lines. Furthermore, this depletion of NICD was associated with significant growth inhibition of all three ovarian cancer cell lines. CONCLUSIONS.: NICD was frequently expressed in ovarian cancer cell lines and human ovarian cancer specimens. Importantly, depletion of Notch 1 led to growth inhibition of ovarian cancer cells. These findings support the hypothesis that Notch 1 plays a role in ovarian cancer proliferation, encouraging the investigation of this pathway as a therapeutic target.


Spontaneous rupture of the left renal collecting system during pregnancy.
Spontaneous rupture of a normal renal collecting system during pregnancy is uncommon and all reported cases have occurred in right kidneys. We report a case of spontaneous rupture of the left renal collecting system during pregnancy. A 33-year-old pregnant woman presented with left loin and lower abdominal pain, and signs of preterm labour, at 32 weeks' gestation. An emergency caesarean section was performed for foetal distress but the left loin pain did not subside after delivery. Ultrasonography and a computed tomogram showed a left perinephric collection and urine extravasation, compatible with rupture of the renal collecting system. A percutaneous nephrostomy was inserted and the symptoms subsided. A summary of the literature discussing management of this clinical situation is provided.


Structural and mutational analyses of Aes, an inhibitor of MalT in Escherichia coli.
The acyl esterase Aes effectively inhibits the transcriptional activity of MalT-the central activator of maltose and maltodextrin utilizing genes in Escherichia coli. To provide better insight into the nature of the interaction between Aes and MalT, we determined two different crystal structures of Aes-in its native form and covalently modified by a phenylmethylsulfonyl moiety at its active site serine. Both structures show distinct space groups and were refined to a resolution of 1.8 Å and 2.3 Å, respectively. The overall structure of Aes resembles a canonical α/β-hydrolase fold, which is extended by a funnel-like cap structure that forms the substrate-binding site. The catalytic triad of Aes, comprising residues Ser165, His292, and Asp262, is located at the bottom of this funnel. Analysis of the crystal-packing contacts of the two different space groups as well as analytical size-exclusion chromatography revealed a homodimeric arrangement of Aes. The Aes dimer adopts an antiparallel contact involving both the hydrolase core and the cap, with its twofold axis perpendicular to the largest dimension of Aes. To identify the surface area of Aes that is responsible for the interaction with MalT, we performed a structure-based alanine-scanning mutagenesis to pinpoint Aes residues that are significantly impaired in MalT inhibition, but still exhibit wild-type expression and enzymatic activity. These residues map to a shallow slightly concave surface patch of Aes at the opposite site of the dimerization interface and indicate the surface area that interacts with MalT.


Relationship of serum immunoglobulin and IgG subclass levels to race, ethnicity and behavioral characteristics in HIV infection.
To determine whether demographic and behavioral factors affect immunoglobulin regulation in HIV infection, we studied injection drug users, women, and minority ethnic and racial groups with and without HIV infection. A prospective cross-sectional study of ambulatory persons with or at risk for HIV infection was conducted. We enrolled 48 injection drug users (IDUs) and 43 non-IDUs seropositive for HIV and 22 seronegative at-risk individuals in the Bronx, New York City. Sixteen HIV-seronegative, non-IDUs controls were also studied. Total serum immunoglobulin levels, IgG subclasses and lymphocyte phenotypes were measured. Serum IgG, IgA, IgG(1) and IgG(3) were increased in all stages of HIV infection controlling for injection drug use, gender, race and age (p0.05). Serum IgM levels were significantly decreased in HIV seropositives compared to HIV seronegatives (p<0.02). Two patterns of serum immunoglobulin level elevation were found in HIV infection: 1) IgG, IgG(1) and IgG(3) levels were elevated in early and advanced HIV infection; 2) IgA, IgG(2) and IgG(4) levels were elevated only in advanced HIV infection. IgG levels were increased in Blacks compared to Caucasians with HIV infection (p=0.01). Serum IgG, IgG(1) and IgG(3) levels are increased in early HIV infection, while serum IgA, IgG(2), and IgG(3) levels are increased only in advanced HIV infection. In contrast, serum IgM levels are decreased in HIV infection. HIV-seropositive Blacks have higher serum IgG levels than HIV-seropositive Caucasians. Further studies are necessary to determine the mechanism(s) underlying the different patterns of immunoglobulin elevation in HIV infection.


Clinicians' Guide to Cannabidiol and Hemp Oils.
Cannabidiol (CBD) oils are low tetrahydrocannabinol products derived from Cannabis sativa that have become very popular over the past few years. Patients report relief for a variety of conditions, particularly pain, without the intoxicating adverse effects of medical marijuana. In June 2018, the first CBD-based drug, Epidiolex, was approved by the US Food and Drug Administration for treatment of rare, severe epilepsy, further putting the spotlight on CBD and hemp oils. There is a growing body of preclinical and clinical evidence to support use of CBD oils for many conditions, suggesting its potential role as another option for treating challenging chronic pain or opioid addiction. Care must be taken when directing patients toward CBD products because there is little regulation, and studies have found inaccurate labeling of CBD and tetrahydrocannabinol quantities. This article provides an overview of the scientific work on cannabinoids, CBD, and hemp oil and the distinction between marijuana, hemp, and the different components of CBD and hemp oil products. We summarize the current legal status of CBD and hemp oils in the United States and provide a guide to identifying higher-quality products so that clinicians can advise their patients on the safest and most evidence-based formulations. This review is based on a PubMed search using the terms CBD, cannabidiol, hemp oil, and medical marijuana. Articles were screened for relevance, and those with the most up-to-date information were selected for inclusion.


Influence of Local Heating on Marangoni Flows and Evaporation Kinetics of Pure Water Drops.
The effect of localized heating on the evaporation of pure sessile water drops was probed experimentally by a combination of infrared thermography and optical imaging. In particular, we studied the effect of three different heating powers and two different locations, directly below the center and edge of the drop. In all cases, four distinct stages were identified according to the emerging thermal patterns. In particular, depending on heating location, recirculating vortices emerge that either remain pinned or move azimuthally within the drop. Eventually, these vortices oscillate in different modes depending on heating location. Infrared data allowed extraction of temperature distribution on each drop surface. In turn, the flow velocity in each case was calculated and was found to be higher for edge heating, due to the one-directional nature of the heating. Additionally, calculation of the dimensionless Marangoni and Rayleigh numbers yielded the prevalence of Marangoni convection. Heating the water drops also affected the evaporation kinetics by promoting the "stick-slip" regime. Moreover, both the total number of depinning events and the pinning strength were found to be highly dependent on heating location. Lastly, we report a higher than predicted relationship between evaporation rate and heating temperature, due to the added influence of the recirculating flows on temperature distribution and hence evaporation flux.


Determination of the protonation preferences of bilin pigments in cryptophyte antenna complexes.
The light-harvesting mechanisms of cryptophyte antenna complexes have attracted considerable attention due to their ability to exhibit maximal photosynthetic activity under very low-light conditions and to display several colors, as well as the observation of vibronic coherent features in their two-dimensional electronic spectra. However, detailed investigations on the interplay between the protein environment and their light-harvesting properties are hampered by the uncertainty related to the protonation state of the underlying bilin pigments. Here we study the protonation preferences of four types of bilin pigments including 15,16-dihydrobiliverdin (DBV), phycoerythrobilin (PEB), phycocyanobilin (PCB) and mesobiliverdin (MBV), which are found in phycoerythrin PE545 and phycocyanin PC577, PC612, PC630 and PC645 complexes. We apply quantum chemical calculations coupled to continuum solvation calculations to predict the intrinsic acidity of bilins in aqueous solution, and then combine molecular dynamics simulations with empirical pKa estimates to investigate the impact of the local protein environment on the acidity of the pigments. We also report measurements of the absorption spectra of the five complexes in a wide range of pH in order to validate our simulations and investigate possible changes in the light harvesting properties of the complexes in the range of physiological pH found in the lumen (pH ∼ 5-7). The results suggest a pKa > 7 for DBV and MBV pigments in the α polypeptide chains of PE545 and PC630/PC645 complexes, which are not coordinated to a negatively charged amino acid. For the other PEB, DBV and PCB pigments, which interact with a Glu or Asp side chain, higher pKa values (pKa > 8) are estimated. Overall, the results support a preferential population of the fully protonated state for bilins in cryptophyte complexes under physiological conditions regardless of the specific type of pigment and local protein environment.


Global shape processing involves feature-selective and feature-agnostic coding mechanisms.
Recent research and modeling proposes that a closed shape is accurately described by both the curvature and angular location of its parts relative to the shape center, implying that the shape is coded along with its overall orientation. We tested this proposition. Radial frequency (RF) patterns were employed as stimuli as they can represent a range of familiar closed shapes and are processed globally. We measured a RF amplitude aftereffect (RFAAE) as a function of the shape orientation difference between adapt and test patterns of the same RF. For RF3 and RF4, RFAAEs were largest when adapt and test patterns were the same orientation, and then linearly decreased as the adaptor was rotated away from the test. RFAAEs did not, however, reach zero, instead plateauing significantly above zero. On the other hand, when adapt and test were of opposite luminance polarity, RFAAEs, although lower than same luminance-polarity RFAAEs, were invariant to differences between adapt and test orientations. Our findings provide evidence for two global shape mechanisms: one that is selective for shape orientation and luminance polarity, and one that is agnostic to these characteristics.


Effects of imperfect test sensitivity and specificity on observational studies of influenza vaccine effectiveness.
The recently developed test-negative design is now standard for observational studies of influenza vaccine effectiveness (VE). It is unclear how influenza test misclassification biases test-negative VE estimates relative to VE estimates from traditional cohort or case-control studies. We simulated populations whose members may develop acute respiratory illness (ARI) due to influenza and to non-influenza pathogens. In these simulations, vaccination reduces the risk of influenza but not of non-influenza ARI. Influenza test sensitivity and specificity, risks of influenza and non-influenza ARI, and VE were varied across the simulations. In each simulation, we estimated influenza VE using a cohort design, a case-control design, and a test-negative design. In the absence of influenza test misclassification, all three designs accurately estimated influenza VE. In the presence of misclassification, all three designs underestimated VE. Bias in VE estimates was slightly greater in the test-negative design than in cohort or case-control designs. Assuming the use of highly sensitive and specific reverse-transcriptase polymerase chain reaction tests for influenza, bias in the test-negative studies was trivial across a wide range of realistic values for VE. Although influenza test misclassification causes more bias in test-negative studies than in traditional cohort or case-control studies, the difference is trivial for realistic combinations of attack rates, test sensitivity/specificity, and VE.


Synthesis and release of human cartilage matrix proteoglycans are differently regulated by nitric oxide and prostaglandin-E2.
Recent studies showed beneficial effects of COX-2 inhibition on proteoglycan turnover of both IL-1beta/tumour necrosis factor alpha (TNFalpha) damaged cartilage and of osteoarthritic cartilage. Although proteoglycan release and content were normalised, proteoglycan synthesis was only partially influenced. Prostaglandin-E2 is the main product formed by COX-2. We therefore evaluate the role of prostaglandin-E2 in relation to nitric oxide in disturbing cartilage proteoglycan turnover. Human healthy cartilage, alone or in the presence of IL-1beta+TNFalpha, was cultured for 7 days with or without prostaglandin-E2 or the selective COX-2 inhibitor (celecoxib 10 microM). Changes in cartilage matrix proteoglycan turnover, levels of prostaglandin-E2 and nitric oxide were determined. Proteoglycan synthesis and release of the cartilage were not affected by prostaglandin-E2 alone. Addition of IL-1beta+TNFalpha to healthy cartilage resulted in inhibition of proteoglycan synthesis and increase in proteoglycan release. When prostaglandin-E2 was added, in addition to IL-1beta+TNFalpha, proteoglycan release increased further, but proteoglycan synthesis was not influenced further. Addition of a selective COX-2 inhibitor to the IL-1beta+TNFalpha treated cartilage inhibited the enhanced prostaglandin-E2 production and almost completely normalised proteoglycan release, whereas synthesis remained unaffected. Also, the enhanced NO-levels remained elevated. Prostaglandin-E2 levels correlated significantly with proteoglycan release, whereas NO levels correlated significantly with proteoglycan synthesis. The present results suggest involvement of prostaglandin-E2 in enhanced cartilage proteoglycan release but not synthesis, although healthy cartilage has to be sensitised by IL-1beta+tumour necrosis factor alpha (TNFalpha). IL-1beta+TNFalpha induced NO seems to be involved in inhibition of proteoglycan synthesis, independent of prostaglandin-E2, and thus seems insensitive to regulation by (selective) COX-2 inhibitors.


Macromolecular Crowding In Vitro, In Vivo, and In Between.
Biochemical processes take place in heterogeneous and highly volume-occupied or crowded environments that can considerably influence the reactivity and distribution of participating macromolecules. We summarize here the thermodynamic consequences of excluded-volume and long-range nonspecific intermolecular interactions for macromolecular reactions in volume-occupied media. In addition, we summarize and compare the information content of studies of crowding in vitro and in vivo. We emphasize the importance of characterizing the behavior not only of labeled tracer macromolecules but also the composition and behavior of unlabeled macromolecules in the immediate vicinity of the tracer. Finally, we propose strategies for extending quantitative analyses of crowding in simple model systems to increasingly complex media up to and including intact cells.


Incidence and types of preceding and subsequent fractures in cats with patellar fracture and dental anomaly syndrome.
The aim of this study was to document the incidence of preceding and subsequent fractures to the patellar fractures in cats with patellar fractures and dental anomaly syndrome. Records of cats with patellar fracture and dental anomaly syndrome were retrieved from the combined databases at the University of Bristol, UK, and Exclusively Cats Veterinary Hospital, USA. A request was made to complete a questionnaire to obtain long-term follow-up of these cats with respect to their current status and fractures to other bones; radiographs and histories were requested and were reviewed for treatment of ongoing fractures and outcome. Of the 191 cases reported with this syndrome, 92 cats (48.2%) had dental anomalies and 78 (40.8%) had fractures to other bones; 21 cats sustained the fractures preceding the patellar fractures and 57 subsequently. In total, there were 175 fractures: acetabulum (25%), tibia (22%), ischium (15.4%), humeral condyle (13.7%), calcaneus (5.1%), ilium (5.1%), pubis (3.4%) and other bones (10.2%). The majority of these fractures were characteristic of insufficiency (stress) fractures with a very similar configuration in each bone. A high proportion of cats with patellar fracture and dental anomaly syndrome will have preceding or subsequent fractures to their patellar fractures. In this study, >10% of cats suffered characteristic fractures preceding the patellar fractures. The presence of these fractures should alert the veterinarian to the possibility that the cat is affected by patellar fracture and dental anomaly syndrome.


A lipid bound actin meshwork organizes liquid phase separation in model membranes.
The eukaryotic cell membrane is connected to a dense actin rich cortex. We present FCS and STED experiments showing that dense membrane bound actin networks have severe influence on lipid phase separation. A minimal actin cortex was bound to a supported lipid bilayer via biotinylated lipid streptavidin complexes (pinning sites). In general, actin binding to ternary membranes prevented macroscopic liquid-ordered and liquid-disordered domain formation, even at low temperature. Instead, depending on the type of pinning lipid, an actin correlated multi-domain pattern was observed. FCS measurements revealed hindered diffusion of lipids in the presence of an actin network. To explain our experimental findings, a new simulation model is proposed, in which the membrane composition, the membrane curvature, and the actin pinning sites are all coupled. Our results reveal a mechanism how cells may prevent macroscopic demixing of their membrane components, while at the same time regulate the local membrane composition. DOI: http://dx.doi.org/10.7554/eLife.01671.001.


Gut Microbiome-Wide Search for Bacterial Azoreductases Reveals Potentially Uncharacterized Azoreductases Encoded in the Human Gut Microbiome.
The human gut is home to trillions of microorganisms that are responsible for the modification of many orally administered drugs, leading to a wide range of therapeutic outcomes. Prodrugs bearing an azo bond are designed to treat inflammatory bowel disease and colorectal cancer via microbial azo reduction, allowing for topical application of therapeutic moieties to the diseased tissue in the intestines. Despite the inextricable link between microbial azo reduction and the efficacy of azo prodrugs, the prevalence, abundance, and distribution of azoreductases have not been systematically examined across the gut microbiome. Here, we curated and clustered amino acid sequences of experimentally confirmed bacterial azoreductases and conducted a hidden Markov model-driven homolog search for these enzymes across 4644 genome sequences present in the representative Unified Human Gastrointestinal Genomes collection. We identified 1958 putative azo-reducing species, corroborating previous findings that azo reduction appears to be a ubiquitous function of the gut microbiome. However, through a systematic comparison of predicted and confirmed azo-reducing strains, we hypothesize the presence of uncharacterized azoreductases in 25 prominent strains of the human gut microbiome. Finally, we confirmed the azo reduction of Acid Orange 7 by multiple strains of Fusobacterium nucleatum, Bacteroides fragilis, and Clostridium clostridioforme Together, these results suggest the presence and activity of many uncharacterized azoreductases in the human gut microbiome and motivate future studies aimed at characterizing azoreductase genes in prominent members of the human gut microbiome. SIGNIFICANCE STATEMENT: This work systematically examined the prevalence, abundance, and distribution of azoreductases across the healthy and inflammatory bowel disease human gut microbiome, revealing potentially uncharacterized azoreductase genes. It also confirmed the reduction of Acid Orange 7 by strains of Fusobacterium nucleatum, Bacteroides fragilis, and Clostridium clostridioforme.


In Vitro Study of Deuterium Effect on Biological Properties of Human Cultured Adipose-Derived Stem Cells.
In current in vitro study we have shown the impact of deuterium content in growth medium on proliferation rate of human cultured adipose-derived stem cells (ADSC). ADSCs have also demonstrated morphological changes when cultured in deuterated growth medium: the cell cultures did not reach confluence but acquired polygonal morphology with pronounced stress fibers. At high deuterium concentrations the ADSCs population doubling time increased which indicated the cell cycle retardation and decrease of cell proliferation rate. The deuterated and deuterium-depleted growth media demonstrated acute and chronic cytotoxicity, respectively. The minimal migration ability was observed in deuterated medium whereas the highest migration activity was observed in the medium with the deuterium content close to natural. The cells in deuterated growth medium demonstrated decrease in metabolic activity after three days in culture. In contrast, in deuterium-depleted medium there was an increase in ADSC metabolic activity.


Preneoplastic prostate lesions: an opportunity for prostate cancer prevention.
Environmental factors, especially the diet, play a prominent role in the epidemic of prostate cancer (PCA), in the United States. Many candidate dietary components have been proposed to influence human prostatic carcinogenesis, including fat, calories, fruits and vegetables, anti-oxidants, and various micronutrients, but the specific roles dietary agents play in promoting or preventing PCA remain controversial. We have collected evidence to suggest that GSTP1, the gene encoding the pi-class glutathione S-transferase (GST), may serve a "caretaker" function for prostatic cells. Although GSTP1 can be detected in normal prostatic epithelium, in almost all PCA cases, PCA cells fail to express GSTP1 polypeptides, and lack of GSTP1 expression most often appears to be the result of somatic "CpG island" DNA methylation changes. Loss of GSTP1 function also appears to be characteristic of prostatic epithelial neoplasia (PIN) lesions, thought to represent PCA precursors. We have recently learned that a new candidate early PCA precursor lesion, proliferative inflammatory atrophy (PIA), characterized by proliferating prostatic cells juxtaposed to inflammatory cells, contains epithelial cells that express high levels of GSTP1. These findings have formed the basis for a new model of prostatic carcinogenesis, in which prostatic cells in PIA lesions, subjected to a barrage of inflammatory oxidants, induce GSTP1 expression as a defense against oxidative genome damage. When cells with defective GSTP1 genes appear amongst the PIA cells, such cells become vulnerable to oxidants and electrophiles that inflict genome damage that tends to promote neoplastic transformation to PIN and PCA cells. Subsequently, PIN and PCA cells with defective GSTPI genes remain vulnerable to similar stresses tending to promote malignant progression. This new model for prostatic carcinogenesis has implications for the design of new prostate cancer prevention strategies. Rational prevention approaches might include: (i) restoration of GSTPI expression via treatment with inhibitors of CpG methylation, (ii) compensation for inadequate GSTPI activity via treatment with inducers of general GST activity, and (iii) abrogation of genome-damaging stresses via avoidance of exogenous carcinogens and/or reduction of endogenous carcinogenic (particularly oxidant) stresses.


The Neurogenic Potential of Astrocytes Is Regulated by Inflammatory Signals.
Although the adult brain contains neural stem cells (NSCs) that generate new neurons throughout life, these astrocyte-like populations are restricted to two discrete niches. Despite their terminally differentiated phenotype, adult parenchymal astrocytes can re-acquire NSC-like characteristics following injury, and as such, these 'reactive' astrocytes offer an alternative source of cells for central nervous system (CNS) repair following injury or disease. At present, the mechanisms that regulate the potential of different types of astrocytes are poorly understood. We used in vitro and ex vivo astrocytes to identify candidate pathways important for regulation of astrocyte potential. Using in vitro neural progenitor cell (NPC)-derived astrocytes, we found that exposure of more lineage-restricted astrocytes to either tumor necrosis factor alpha (TNF-α) (via nuclear factor-κB (NFκB)) or the bone morphogenetic protein (BMP) inhibitor, noggin, led to re-acquisition of NPC properties accompanied by transcriptomic and epigenetic changes consistent with a more neurogenic, NPC-like state. Comparative analyses of microarray data from in vitro-derived and ex vivo postnatal parenchymal astrocytes identified several common pathways and upstream regulators associated with inflammation (including transforming growth factor (TGF)-β1 and peroxisome proliferator-activated receptor gamma (PPARγ)) and cell cycle control (including TP53) as candidate regulators of astrocyte phenotype and potential. We propose that inflammatory signalling may control the normal, progressive restriction in potential of differentiating astrocytes as well as under reactive conditions and represent future targets for therapies to harness the latent neurogenic capacity of parenchymal astrocytes.


Predicting violent infractions in a Swiss state penitentiary: a replication study of the PCL-R in a population of sex and violent offenders.
Research conducted with forensic psychiatric patients found moderate correlations between violence in institutions and psychopathy. It is unclear though, whether the PCL-R is an accurate instrument for predicting aggressive behavior in prisons. Results seem to indicate that the instrument is better suited for predicting verbal rather than physical aggression of prison inmates. PCL-R scores were assessed for a sample of 113 imprisoned sex and violent offenders in Switzerland. Logistic regression analyses were used to estimate physical and verbal aggression as a function of the PCL-R sum score. Additionally, stratified analyses were conducted for Factor 1 and 2. Infractions were analyzed as to their motives and consequences. The mean score of the PCL-R was 12 points. Neither the relationship between physical aggression and the sum score of the PCL-R, nor the relationship between physical aggression and either of the two factors of the PCL-R were significant. Both the sum score and Factor 1 predicted the occurrence of verbal aggression (AUC=0.70 and 0.69), while Factor 2 did not. Possible explanations are discussed for the weak relationship between PCL-R scores and physically aggressive behavior during imprisonment. Some authors have discussed whether the low base rate of violent infractions can be considered an explanation for the non-significant relation between PCL-R-score and violence. The base rate in this study, however, with 27%, was not low. It is proposed that the distinction between reactive and instrumental motives of institutional violence must be considered when examining the usefulness of the PCL-R in predicting in-prison physical aggressive behavior.


Inactivating anterior insular cortex reduces risk taking.
We often have to make risky decisions between alternatives with outcomes that can be better or worse than the outcomes of safer alternatives. Although previous studies have implicated various brain regions in risky decision making, it remains unknown which regions are crucial for balancing whether to take a risk or play it safe. Here, we focused on the anterior insular cortex (AIC), the causal involvement of which in risky decision making is still unclear, although human imaging studies have reported AIC activation in various gambling tasks. We investigated the effects of temporarily inactivating the AIC on rats' risk preference in two types of gambling tasks, one in which risk arose in reward amount and one in which it arose in reward delay. As a control within the same subjects, we inactivated the adjacent orbitofrontal cortex (OFC), which is well known to affect risk preference. In both gambling tasks, AIC inactivation decreased risk preference whereas OFC inactivation increased it. In risk-free control situations, AIC and OFC inactivations did not affect decision making. These results suggest that the AIC is causally involved in risky decision making and promotes risk taking. The AIC and OFC may be crucial for the opposing motives of whether to take a risk or avoid it.


Precise transverse alignment of spatial light modulator sections for complex optical field generation.
Based on the properties of the dove prism and the Fourier optics approach, the coordinate relationships among four spatial light modulator (SLM) sections in a vectorial optical field generator are derived and experimentally verified. Taking the coordinate system of the first SLM section as a reference, the coordinate displacements between the first section and subsequent sections are determined via employing specially designed four-quadrant patterns, which enable the visualization of the degree of freedom controlled by each SLM section. A complex optical field could be accurately generated through combining the derived coordinate relationships and pre-compensation of the measured coordinate displacements. Several typical complex optical fields are experimentally generated to demonstrate the validity of the proposed transverse alignment method.


The mechanism of 5-methyltetrahydrofolate transport by human erythrocytes.
The mechanism involved in 5-methyltetrahydrofolate uptake by human cells is poorly understood. To more clearly elucidate this physiologically important process, transport of the vitamin was studied in human erythrocytes. 5-methyltetrahydrofolate uptake was found to increase with reticulocytosis, but measurable incorporation occurred in erythrocyte suspensions depleted of reticulocytes, leukocytes, and platelets, indicating uptake by mature erythrocytes. Incubation of erythrocytes with increasing concentrations of [(14)C]5-methyltetrahydrofolate resulted in increasing uptake but decreasing percentage incorporation, consistent with saturation of a carrier system. Both influx and efflux phases of uptake were temperature dependent, with almost no transport at 4 degrees C. Uptake of [(14)C]5-methytetrahydrofolate was effectively inhibited by unlabeled 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, and methotrexate, but not by pteroylglutamic acid. Prior incubation with 5-formyltetrahydrofolate increased uptake of [(14)C]5-methyltetrahydrofolate, and extracellular 5-formyltetrahydrofolate enhanced efflux of [(14)C]5-methyltetrahydrofolate. Nearly total depletion of ATP increased uptake of [(14)C]5-methyltetrahydrofolate, but efflux was unchanged. Column chromatography of membrane-free hemolysate after incubation with [(14)C]5-methyltetrahydrofolate showed 95% of radioactivity corresponded to marker radioisotope, and no other peak was noted. Thus peripheral erythrocytes incorporate 5-methyltetrahydrofolate by a saturable, temperature-dependent, substrate-specific process which is influenced by counter-transport. This mechanism is qualitatively similar to the carrier-mediated transport of folate compounds previously described in other cell types. Therefore, human erythrocytes should be useful for detailed characterization of this membrane carrier system.


A Robust Mammary Organoid System to Model Lactation and Involution-like Processes.
The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women worldwide. Studying the regulatory mechanisms of mammary gland development is essential for understanding how dysregulation can lead to breast cancer initiation and progression. Three-dimensional (3D) mammary organoids offer many exciting possibilities for the study of tissue development and breast cancer. In the present protocol derived from Sumbal et al., we describe a straightforward 3D organoid system for the study of lactation and involution ex vivo. We use primary and passaged mouse mammary organoids stimulated with fibroblast growth factor 2 (FGF2) and prolactin to model the three cycles of mouse mammary gland lactation and involution processes. This 3D organoid model represents a valuable tool to study late postnatal mammary gland development and breast cancer, in particular postpartum-associated breast cancer. Graphic abstract: Mammary gland organoid isolation and culture procedures.


Fast Dissolving Sublingual Films of Ondansetron Hydrochloride: Effect of Additives on in vitro Drug Release and Mucosal Permeation.
Ondansetron hydrochloride, a 5 HT3 antagonist is a powerful antiemetic drug which has oral bioavailability of 60% due to hepatic first pass metabolism and has a short half-life of 5 h. To overcome the above draw back, the present study was carried out to formulate and evaluate fast dissolving films of ondansetron hydrochloride for sublingual administration. The films were prepared from polymers such as polyvinylalcohol, polyvinyl pyrrolidone, Carbopol 934P in different ratios by solvent casting method. Propylene glycol or PEG 400 as plasticizers and mannitol or sodium saccharin as sweeteners were also included. The IR spectral studies showed no interaction between drug and polymer or with other additives. Satisfactory results were obtained when subjected to physico-chemical tests such as uniformity of weight, thickness, surface pH, folding endurance, uniformity of drug content, swelling index, bioadhesive strength, and tensile strength. Films were also subjected to in vitro drug release studies by using USP dissolution apparatus. Ex vivo drug permeation studies were carried out using porcine membrane model. In vitro release studies indicated 81-96% release within 7 min and 66-80% within 7 min during ex vivo studies. Drug permeation of 66-77% was observed through porcine mucosa within 40 min. Higher percentage of drug release was observed from films containing the sweeteners. The stability studies conducted for a period of 8 weeks showed no appreciable change in drug content, surface pH, and in vitro drug release.


Tremor-ataxia with central hypomyelination (TACH) leukodystrophy maps to chromosome 10q22.3-10q23.31.
Leukodystrophies are a heterogeneous group of disorders associated with abnormal central nervous system white matter. The clinical features invariably include upper motor neuron signs and developmental regression with or without other neurological manifestations. The objective of this study was to characterize clinically and genetically a new form of childhood-onset leukodystrophy with ataxia and tremor. We recruited seven French-Canadian cases belonging to five families affected by an unknown form of childhood-onset leukodystrophy. Genome-wide scans (GWS) were performed using the Illumina Hap310 or Hap610 Bead Chip to identify regions of shared homozygosity that were further studied for linkage with STS markers. All cases presented between the ages of 1 and 5 years with spasticity along with other upper motor neuron signs, prominent postural tremor, and cerebellar signs. Though motor regression is a constant feature, cognitive functions are relatively preserved, even late in the course of the disease. The higher frequency of founder diseases in the French-Canadian population and the segregation in pedigrees are suggestive of a recessive mode of inheritance. By homozygosity mapping, we established linkage to a 12.6-Mb SNP-haplotyped region on chromosome 10q22.3-10q23.31 (maximum LOD score: 5.47). We describe an autosomal recessive childhood-onset leukodystrophy with ataxia and tremor mapping to a 12.6 Mb interval on chromosome 10q22.3-10q23.31. Identification of the mutated gene will allow precise diagnosis and genetic counseling and shed light on how its perturbed function leads to white matter abnormalities.


Why do patients with multimorbidity in England report worse experiences in primary care? Evidence from the General Practice Patient Survey.
To describe and explain the primary care experiences of people with multiple long-term conditions in England. Using questionnaire data from 906,578 responders to the English 2012 General Practice Patient Survey, we describe the primary care experiences of patients with long-term conditions, including 583,143 patients who reported one or more long-term conditions. We employed mixed effect logistic regressions to analyse data on six items covering three care domains (access, continuity and communication) and a single item on overall primary care experience. We controlled for sociodemographic characteristics, and for general practice using a random effect, and further, controlled for, and explored the importance of, health-related quality of life measured using the EuroQoL (EQ-5D) scale. Most patients with long-term conditions report a positive experience of care at their general practice (after adjusting for sociodemographic characteristics and general practice, range 74.0-93.1% reporting positive experience of care across seven questions) with only modest variation by type of condition. For all three domains of patient experience, an increasing number of comorbid conditions is associated with a reducing percentage of patients reporting a positive experience of care. For example, compared with respondents with no long-term condition, the OR for reporting a positive experience is 0.83 (95% CI 0.80 to 0.87) for respondents with four or more long-term conditions. However, this relationship is no longer observed after adjusting for health-related quality of life (OR (95% CI) single condition=1.23 (1.21 to 1.26); four or more conditions=1.31 (1.25 to 1.37)), with pain making the greatest difference among five quality of life variables included in the analysis. Patients with multiple long-term conditions more frequently report worse experiences in primary care. However, patient-centred measures of health-related quality of life, especially pain, are more important than the number of conditions in explaining why patients with multiple long-term conditions report worse experiences of care.


Droplet impact on heated powder bed.
The impact of droplets on a heated powder bed involves a wide range of phenomena with increasing complexity from spreading of liquid, to bubble nucleation, to more complex ones such as splashing, crater formation, and fluidization. In this study, we focus on the impact behaviors caused by elevation in temperature of the powder bed. We experimentally characterize the impact behaviors for wide ranges of the impact velocity and surface temperature of the powder bed. We reveal several phenomena specific to the impact on a heated powder bed such as rebound, ruptures of expanding liquid films, bubble nucleation, and fluidization. We also show that the maximum spreading diameter increases with the surface temperature of the powder bed and propose an empirical scaling law to describe such a relation.


The World Health Organization Fetal Growth Charts: A Multinational Longitudinal Study of Ultrasound Biometric Measurements and Estimated Fetal Weight.
Perinatal mortality and morbidity continue to be major global health challenges strongly associated with prematurity and reduced fetal growth, an issue of further interest given the mounting evidence that fetal growth in general is linked to degrees of risk of common noncommunicable diseases in adulthood. Against this background, WHO made it a high priority to provide the present fetal growth charts for estimated fetal weight (EFW) and common ultrasound biometric measurements intended for worldwide use. We conducted a multinational prospective observational longitudinal study of fetal growth in low-risk singleton pregnancies of women of high or middle socioeconomic status and without known environmental constraints on fetal growth. Centers in ten countries (Argentina, Brazil, Democratic Republic of the Congo, Denmark, Egypt, France, Germany, India, Norway, and Thailand) recruited participants who had reliable information on last menstrual period and gestational age confirmed by crown-rump length measured at 8-13 wk of gestation. Participants had anthropometric and nutritional assessments and seven scheduled ultrasound examinations during pregnancy. Fifty-two participants withdrew consent, and 1,387 participated in the study. At study entry, median maternal age was 28 y (interquartile range [IQR] 25-31), median height was 162 cm (IQR 157-168), median weight was 61 kg (IQR 55-68), 58% of the women were nulliparous, and median daily caloric intake was 1,840 cal (IQR 1,487-2,222). The median pregnancy duration was 39 wk (IQR 38-40) although there were significant differences between countries, the largest difference being 12 d (95% CI 8-16). The median birthweight was 3,300 g (IQR 2,980-3,615). There were differences in birthweight between countries, e.g., India had significantly smaller neonates than the other countries, even after adjusting for gestational age. Thirty-one women had a miscarriage, and three fetuses had intrauterine death. The 8,203 sets of ultrasound measurements were scrutinized for outliers and leverage points, and those measurements taken at 14 to 40 wk were selected for analysis. A total of 7,924 sets of ultrasound measurements were analyzed by quantile regression to establish longitudinal reference intervals for fetal head circumference, biparietal diameter, humerus length, abdominal circumference, femur length and its ratio with head circumference and with biparietal diameter, and EFW. There was asymmetric distribution of growth of EFW: a slightly wider distribution among the lower percentiles during early weeks shifted to a notably expanded distribution of the higher percentiles in late pregnancy. Male fetuses were larger than female fetuses as measured by EFW, but the disparity was smaller in the lower quantiles of the distribution (3.5%) and larger in the upper quantiles (4.5%). Maternal age and maternal height were associated with a positive effect on EFW, particularly in the lower tail of the distribution, of the order of 2% to 3% for each additional 10 y of age of the mother and 1% to 2% for each additional 10 cm of height. Maternal weight was associated with a small positive effect on EFW, especially in the higher tail of the distribution, of the order of 1.0% to 1.5% for each additional 10 kg of bodyweight of the mother. Parous women had heavier fetuses than nulliparous women, with the disparity being greater in the lower quantiles of the distribution, of the order of 1% to 1.5%, and diminishing in the upper quantiles. There were also significant differences in growth of EFW between countries. In spite of the multinational nature of the study, sample size is a limiting factor for generalization of the charts. This study provides WHO fetal growth charts for EFW and common ultrasound biometric measurements, and shows variation between different parts of the world.


Uptake, recognition and responses to peptidoglycan in the mammalian host.
Microbiota, and the plethora of signalling molecules that they generate, are a major driving force that underlies a striking range of inter-individual physioanatomic and behavioural consequences for the host organism. Among the bacterial effectors, one finds peptidoglycan, the major constituent of the bacterial cell surface. In the steady-state, fragments of peptidoglycan are constitutively liberated from bacterial members of the gut microbiota, cross the gut epithelial barrier and enter the host system. The fate of these peptidoglycan fragments, and the outcome for the host, depends on the molecular nature of the peptidoglycan, as well the cellular profile of the recipient tissue, mechanism of cell entry, the expression of specific processing and recognition mechanisms by the cell, and the local immune context. At the target level, physiological processes modulated by peptidoglycan are extremely diverse, ranging from immune activation to small molecule metabolism, autophagy and apoptosis. In this review, we bring together a fragmented body of literature on the kinetics and dynamics of peptidoglycan interactions with the mammalian host, explaining how peptidoglycan functions as a signalling molecule in the host under physiological conditions, how it disseminates within the host, and the cellular responses to peptidoglycan.


Recent Progress on the Alloy-Based Anode for Sodium-Ion Batteries and Potassium-Ion Batteries.
High-energy batteries with low cost are urgently needed in the field of large-scale energy storage, such as grid systems and renewable energy sources. Sodium-ion batteries (SIBs) and potassium-ion batteries (PIBs) with alloy-based anodes provide huge potential due to their earth abundance, high capacity, and suitable working potential, and are recognized as attractive alternatives for next-generation batteries system. Although some important breakthroughs have been reported, more significant improvements are still required for long lifetime and high energy density. Herein, the latest progress for alloy-based anodes for SIBs and PIBs is summarized, mainly including Sn, Sb, Ge, Bi, Si, P, and their oxides, sulfides, selenides, and phosphides. Specifically, the material designs for the desired Na+ /K+ storage performance, phase transform, ionic/electronic transport kinetics, and specific chemical interactions are discussed. Typical structural features and research strategies of alloy-based anodes, which are used to facilitate processes in battery development for SIBs and PIBs, are also summarized. The perspective of future research of SIBs and PIBs is outlined.


A Case Report of Transient Cortical Blindness After Angiography.
Transient cortical blindness (TCB) is a rare complication after angiography and is proposed to be related to contrast extravasation. We present the first case of TCB after angiography that was evaluated using serial arterial spin-labeling magnetic resonance perfusion (ASLMRP) and susceptibility-weighted imaging (SWI). A 65-year-old woman with dyslipidemia who had been treated for left vertebral artery dissection 3 months previously presented with dizziness and right hemiparesis. Magnetic resonance imaging of the brain revealed suspected dissection of the V4 segment of the left vertebral artery. Cerebral angiography was performed using a total of 46 mL of nonionic contrast material. Approximately 3 hours after the procedure, the patient started complaining of a slight headache and nausea along with bilateral cortical blindness. The 6-hour ASLMRP and SWI of the brain, respectively, revealed hypoperfusion and increased oxygen extraction fraction in the bilateral occipital lobes. The patient gradually regained vision over the next 12 hours, and follow-up ASLMRP and SWI revealed normal perfusion and normal SWI findings, respectively. Hypoperfusion on ASLMRP and increased oxygen extraction fraction on SWI results derived for a patient presenting with TCB were demonstrated. Thus, we hypothesize that endothelin-related vasoconstriction is a pathophysiologic mechanism for TCB. To the best of our knowledge, this report is the first to describe ASLMRP and SWI findings in TCB after angiography.


Repairing DNA double-strand breaks by the prokaryotic non-homologous end-joining pathway.
The NHEJ (non-homologous end-joining) pathway is one of the major mechanisms for repairing DSBs (double-strand breaks) that occur in genomic DNA. In common with eukaryotic organisms, many prokaryotes possess a conserved NHEJ apparatus that is essential for the repair of DSBs arising in the stationary phase of the cell cycle. Although the bacterial NHEJ complex is much more minimal than its eukaryotic counterpart, both pathways share a number of common mechanistic features. The relative simplicity of the prokaryotic NHEJ complex makes it a tractable model system for investigating the cellular and molecular mechanisms of DSB repair. The present review describes recent advances in our understanding of prokaryotic end-joining, focusing primarily on biochemical, structural and cellular aspects of the mycobacterial NHEJ repair pathway.


Meat eating and nutritional quality of lambs sired by high and low muscle density rams.
Intramuscular fat (IMF) content affects eating and nutritional quality of lamb meat. Muscle density measured by computer tomography is an in vivo proxy measure of IMF content that affects eating and nutritional quality of lamb meat. Lambs sired by high muscle density (HMD) or low muscle density (LMD) rams, selected for slaughter on commercial criteria were measured for meat quality and nutritional traits. A restricted maximum likelihood model was used to compare lamb traits. Additionally, regression analysis of sire estimated breeding value (EBV) for muscle density was performed for each meat quality trait. Muscle density EBV had a negative regression with IMF content (P < 0.001). For each unit increase in muscle density EBV, there was a significant decrease in loin (-1.69 mg/100 g fresh weight) and topside IMF (-0.03 mg/100 g fresh weight). Muscle density EBV had a negative regression with grouped saturated and monounsaturated fatty acids concentration (and monounsaturated proportion P < 0.001). Muscle density EBV had a negative regression with loin sensory traits tenderness, juiciness and overall liking and many novel tenderness sensory traits measured (P < 0.05). Selecting for LMD EBV increased IMF content and favourable meat eating quality traits. In contrast, sire muscle density EBV had a positive regression with loin polyunsaturated:saturated fat ratio and grouped polyunsaturated proportion traits (including total polyunsaturated proportion, total omega-6 (n-6) and total omega-3 (n-3) fatty acids (P < 0.001). This is explained by the fact that as sire muscle density EBV increases, polyunsaturated fatty acid proportion increases and the proportion of saturated and monounsaturated fatty acid content decreases. Muscle density EBV had a positive regression with shear force and the novel toughness sensory traits (P < 0.05). Selection for HMD EBV's increased shear force and toughness traits, which is unfavourable for the consumer. Low muscle density sired meat had higher meat colour traits chroma/saturation (+0.64, SD 2.30, P = 0.012), redness (+0.52, SD 1.91, P = 0.012) and yellowness (+0.31, SD 1.49, P = 0.08) compared to HMD sired meat. Selection for LMD could be used within a breeding programme to increase IMF content and enhance both meat colour and improve eating quality parameters.


Structure-activity relationships in cholapod anion carriers: enhanced transmembrane chloride transport through substituent tuning.
Chloride transport by a series of steroid-based "cholapod" receptors/carriers was studied in vesicles. The principal method involved preincorporation of the cholapods in the vesicle membranes, and the use of lucigenin fluorescence quenching to detect inward-transported Cl-. The results showed a partial correlation between anion affinity and transport activity, in that changes at the steroidal 7 and 12 positions affected both properties in concert. However, changes at the steroidal 3-position yielded irregular effects. Among the new steroids investigated the bis-p-nitrophenylthiourea 3 showed unprecedented activity, giving measurable transport through membranes with a transporter/lipid ratio of 1:250 000 (an average of <2 transporter molecules per vesicle). Increasing transporter lipophilicity had no effect, and positively charged steroids had low activity. The p-nitrophenyl monourea 25 showed modest but significant activity. Measurements using a second method, requiring the addition of transporters to preformed vesicle suspensions, implied that transporter delivery was problematic in some cases. A series of measurements employing membranes of different thicknesses provided further evidence that the cholapods act as mobile anion carriers.


Stress induced cortisol release and schizotypy.
Cortisol is involved in preparing the body's response for stress. However, in those at risk for mental health problems, abnormal cortisol release following stress has been reported. In particular, we are yet to fully understand how stress leads to an exacerbation of symptoms and progression of risk in those who express psychosis proneness or schizotypy. Using the Trier Social Stress Test, we examined the effect of experimentally induced psychosocial stress on cortisol release in otherwise healthy individuals with schizotypal traits. This cross-sectional study included 58 individuals (32.76% male, mean age 22.43). Schizotypy was assessed by total Schizotypal Personality Questionnaire score and we additionally captured ratings of subjective stress. Salivary cortisol was collected over six time points spread prior to and after stress induction and was available for analysis in 39 individuals (28.21% male, mean age 22.77). Those with high schizotypal traits exhibited higher baseline cortisol levels (5.18 nmoL vs 3.71 nmoL). However, those with high schizotypal traits also displayed reduced mean cortisol release (2.02 nmoL vs 5.11 nmoL) and had a delayed cortisol release peak following psychosocial stress. These results indicate those with high schizotypal traits do not display physiological readiness following psychosocial stressors, perhaps due to an already taxed stress system.


Somatic mutations in BRCA1 and BRCA2 could expand the number of patients that benefit from poly (ADP ribose) polymerase inhibitors in ovarian cancer.
The prevalence of BRCA(1/2) mutations in germline DNA from unselected ovarian cancer patients is 11% to 15.3%. It is important to determine the frequency of somatic BRCA(1/2) changes, given the sensitivity of BRCA-mutated cancers to poly (ADP ribose) polymerase-1 (PARP1) inhibitors and platinum analogs. In 235 unselected ovarian cancers, BRCA(1/2) was sequenced in 235, assessed by copy number analysis in 95, and tiling arrays in 65. 113 tumors were sequenced for TP53. BRCA(1/2) transcript levels were assessed by quantitative polymerase chain reaction in 220. When available for tumors with BRCA(1/2) mutations, germline DNA was sequenced. Forty-four mutations (19%) in BRCA1 (n = 31)/BRCA2 (n = 13) were detected, including one homozygous BRCA1 intragenic deletion. BRCA(1/2) mutations were particularly common (23%) in high-grade serous cancers. In 28 patients with available germline DNA, nine (42.9%) of 21 and two (28.6%) of seven BRCA1 and BRCA2 mutations were demonstrated to be somatic, respectively. Five mutations not previously identified in germline DNA were more commonly somatic than germline (four of 11 v one of 17; P = .062). There was a positive association between BRCA1 and TP53 mutations (P = .012). BRCA(1/2) mutations were associated with improved progression-free survival (PFS) after platinum-based chemotherapy in univariate (P = .032; hazard ratio [HR] = 0.65; 95% CI, 0.43 to 0.98) and multivariate (P = .019) analyses. BRCA(1/2) deficiency, defined as BRCA(1/2) mutations or expression loss (in 24 [13.3%] BRCA(1/2)-wild-type cancers), was present in 67 ovarian cancers (30%) and was also significantly associated with PFS in univariate (P = .026; HR = 0.67; 95% CI, 0.47 to 0.96) and multivariate (P = .008) analyses. BRCA(1/2) somatic and germline mutations and expression loss are sufficiently common in ovarian cancer to warrant assessment for prediction of benefit in clinical trials of PARP1 inhibitors.


A longitudinal study on the effects of maternal smoking and secondhand smoke exposure during pregnancy on neonatal neurobehavior.
Maternal smoking during pregnancy is one of the most modifiable causes of morbidity and mortality for both pregnant women and their fetuses. The long-term effects of prenatal exposure to smoke on child behavior and development have been the subject of more extensive research than have the short-term effects. Therefore, the aim of this work is to examine the effects of smoke exposure during pregnancy on neonatal behavior, including in our study a group of mothers exposed to secondhand smoke. The behavior of 282 healthy full-term newborns was assessed using the Neonatal Behavior Assessment Scale (NBAS) at 48-72 h of life. Sixty-two mothers smoked during pregnancy (no mother smoked more than 15 cig/day) and 17 were exposed to secondhand smoke. After adjusting for socio-demographic and obstetric factors, both newborns whose mothers smoked and those whose mothers were exposed to secondhand smoke showed significantly lower scores in the habituation cluster than non-smoking mothers. Exposure to secondhand smoke was also related to lower motor system cluster scores as well as some supplementary items and the newborns of smoking mothers showed significantly lower scores in the state regulation cluster and in some items of the state organization cluster than the newborns of non-smoking mothers. We conclude that active and passive smoking during pregnancy affects several aspects of neurobehavioral development, regardless of socio-demographic, obstetric and pediatric factors.


The developmental increase in ACh current densities on rat sympathetic neurons correlates with changes in nicotinic ACh receptor alpha-subunit gene expression and occurs independent of innervation.
Determining factors that control the expression of neurotransmitter receptors and the mechanisms by which these factors operate is essential to understand how synapses form during development and how receptor numbers change in the adult. In this study, we have investigated one such factor, the influence of innervation, on the developmental expression of nicotinic ACh receptors (nAChRs) on neonatal rat sympathetic neurons, both in terms of ACh current densities, and in terms of mRNA levels for the transcripts that encode these receptors. To date, nine genes have been cloned that encode neuronal nAChRs subunits in mammals. We demonstrate that mRNA encoding five nAChR subunits, alpha 3, alpha 5, alpha 7, beta 2, and beta 4, are present in neonatal rat sympathetic neurons. We show that mRNA levels for alpha 3 and alpha 7 subunits increase by more than threefold over the first 2 postnatal weeks, a period when most synapses are forming on the neurons; however, we observed no significant change in mRNA levels for alpha 5, beta 2, or beta 4. Using whole-cell voltage-clamp techniques, we show that the increase in alpha-subunit mRNA correlates with increases in ACh current densities, which double over the same period. To investigate the role of innervation, we cut the preganglionic nerve at birth and measured subunit mRNA levels and ACh current densities in denervated neurons 1-2 weeks later. Our results indicate that the preganglionic nerve differentially affects the mRNA level for the five nAChR transcripts, yet it has little influence on the developmental increase in ACh current densities.


Inhalation of cold air increases the number of inflammatory cells in the lungs in healthy subjects.
Prolonged exposure to cold air may induce a chronic asthma-like condition in healthy subjects as has been demonstrated in cross-country skiers. In the present controlled study, our aim was to elucidate further the link between cold air exposure and airway inflammation by assessing the cellular influx and mediator levels within the airways following acute exposure to cold air. Bronchoalveolar (BAL) and nasal lavages were performed after exposure to cold air (-23 degrees C) and normal indoor air (+22 degrees C) during a light, intermittent work for 2 h in a cross-over design in eight healthy, nonsmoking, subjects. Analyses of inflammatory cell number, cell activation markers, pro-inflammatory cytokines, albumin and interleukin (IL)-8 in lavage fluids were performed. The number of granulocytes and of alveolar macrophages in BAL fluid was significantly higher after cold air exposure (p<0.05). No increase in BAL fluid lymphocytes and no signs of lymphocyte activation in BAL fluid were found. The concentration of IL-8 was unchanged. There were no signs of granulocyte activation (myeloperoxidase, eosinphilic cationic protein) in BAL fluid. Cold air did not influence the number of inflammatory cells or the concentration of albumin and IL-8 in nasal lavage fluid. In conclusion, exposure to cold air induces an increased number of granulocytes and macrophages in the lower airways in healthy subjects without influencing other inflammatory indices such as cellular activation, plasma leakage and pro-inflammatory cytokines. These findings support the hypothesis that cold air could be of pathogenetic importance in the asthma-like condition previously found in cross-country skiers.


Vitamins C and E inhibit apoptosis of cultured human term placenta trophoblast.
Preeclampsia can be lethal to both mother and baby. The prominent symptoms of this syndrome are hypertension, proteinuria and oedema, resulting from an exaggerated aseptic systemic inflammatory response, triggered by placental factors shed into the maternal circulation. Syncytiotrophoblast microparticles (STBM) are one possible factor, shed when the placenta is exposed to stressors such as hypoxia/reperfusion. These can disrupt mitochondria, triggering apoptosis and necrosis, placental pathologies which are increased in preeclampsia. We tested the effects of antioxidant vitamins C (50 microM) and E (50 microM) on trophoblast in culture, using term villous cytotrophoblast preparations. Following Percoll gradient centrifugation and MHC class I expressing cell depletion of placenta digests, syncytial fragments were removed using anti-placental alkaline phosphatase antibody. This yielded cytotrophoblasts of consistently high purity. EGF (10 ng/ml) stimulated syncytialisation and hCG and progesterone production. However, mitochondrial induced apoptosis (MIA) was evident 96h post-isolation, as mitochondrial membrane potential loss and caspase 9 and caspase 3 activation. ROCK-1 cleavage and syncytiotrophoblast particle shedding increased concurrently with apoptosis induction. Vitamins blocked MIA and syncytiotrophoblast particle shedding and significantly increased hCG (p<0.005) and progesterone (p<0.02) concentrations in culture supernatants, reflecting the increased survival rates. Although more cells survived in culture, syncytialisation rate (%) was significantly reduced (p<0.005). We conclude that vitamins C and E can significantly reduce mitochondrial damage generated following syncytialisation in vitro. However, further work is required to determine whether antioxidant vitamins interfere with normal fusion processes.


Platinum catalyzed sequential hydroboration of decaborane: a facile approach to poly(alkenyldecaborane) with decaborane in the mainchain.
Catalytic sequential hydroboration of decaborane for the synthesis of poly(organodecaborane), with decaborane in the mainchain, is reported for the first time. Under a platinum catalytic system, poly(6-hexenyldecaborane) and poly(6-norbornenyldecaborane) are obtained with well-defined structures and a moderate yield. Thermogravimetric analysis demonstrates that the char yields are 73% and 82%, respectively.


Mechanism of immune complex-mediated damage: induction of interleukin 1 by immune complexes and synergy with interferon-gamma and tumor necrosis factor-alpha.
The ability of immune complexes derived from patients with type II mixed essential cryoglobulinemia to trigger the production of interleukin 1 (IL 1) was investigated. Immune complexes containing either IgM/IgG or IgA/IgG aggregates were shown to induce IL 1 production in human peripheral blood monocyte-enriched populations. Interferon-gamma alone did not induce detectable IL 1, but increased the IL 1 production induced by the immune complexes. Tumor necrosis factor-alpha, which is a potent inducer of IL 1, also enhanced IL 1 production following stimulation with suboptimal doses of the immune complex. The findings suggest that immune complexes may induce inflammation partly due to their capacity to induce the synthesis of IL 1.


A C. elegans model to study human metabolic regulation.
Lipid metabolic disorder is a critical risk factor for metabolic syndrome, triggering debilitating diseases like obesity and diabetes. Both obesity and diabetes are the epicenter of important medical issues, representing a major international public health threat. Accumulation of fat in adipose tissue, muscles and liver and/or the defects in their ability to metabolize fatty acids, results in insulin resistance. This triggers an early pathogenesis of type 2 diabetes (T2D). In mammals, lipid metabolism involves several organs, including the brain, adipose tissue, muscles, liver, and gut. These organs are part of complex homeostatic system and communicate through hormones, neurons and metabolites. Our study dissects the importance of mammalian Krüppel-like factors in over all energy homeostasis. Factors controlling energy metabolism are conserved between mammals and Caenorhabditis elegans providing a new and powerful strategy to delineate the molecular pathways that lead to metabolic disorder. The C. elegans intestine is our model system where genetics, molecular biology, and cell biology are used to identify and understand genes required in fat metabolism. Thus far, we have found an important role of C. elegans KLF in FA biosynthesis, mitochondrial proliferation, lipid secretion, and β-oxidation. The mechanism by which KLF controls these events in lipid metabolism is unknown. We have recently observed that C. elegans KLF-3 selectively acts on insulin components to regulate insulin pathway activity. There are many factors that control energy homeostasis and defects in this control system are implicated in the pathogenesis of human obesity and diabetes. In this review we are discussing a role of KLF in human metabolic regulation.


Multidisciplinary oncolytic virotherapy for gastrointestinal cancer.
Replication-selective tumor-specific viruses represent a novel approach for treating neoplastic diseases. These vectors are designed to induce virus-mediated lysis of tumor cells after selective intracellular virus propagation. For targeting cancer cells, the use of tissue- or cell-specific promoters that are expressed in diverse tumor types but silent in normal cells is required. Human telomerase is highly active in more than 85% of primary cancers, regardless of tissue origin, and its activity is closely correlated with human telomerase reverse transcriptase (hTERT) expression. We constructed an attenuated adenovirus 5 vector (telomelysin, OBP-301) in which the hTERT promoter element drives expression of E1 genes. As only tumor cells that express the telomerase can activate this promoter, the hTERT proximal promoter allows for preferential expression of viral genes in tumor cells, leading to selective viral replication and oncolytic cell death. Upon US Food and Drug Administration approval, a phase 1 dose-escalation study of intratumoral injection of telomelysin for various solid tumors has been completed to confirm the safety, tolerability, and feasibility of the agent. Moreover, we found that adenoviral E1B 55-kDa protein in telomelysin inhibits the radiation-induced DNA repair machinery. Thus, tumor cells infected with telomelysin could be rendered sensitive to ionizing radiation. Recently, we assessed the safety and efficacy of intratumoral injection of telomelysin with radiotherapy in esophageal cancer patients not suited for standard treatments. This review highlights some very promising clinical advances in cancer therapeutic technologies using telomerase-specific oncolytic virotherapy.


Health and health behavior differences: U.S. Military, veteran, and civilian men.
Little is known about health and health behavior differences among military service veterans, active duty service members, National Guard/Reserve members, and civilians. Several important differences were identified among U.S. women from these subpopulations; to identify areas for targeted intervention, studies comparing men from these subpopulations are needed. To compare veteran, military, and civilian men on leading U.S. health indicators. Data were from the 2010 Behavioral Risk Factor Surveillance Survey, a U.S. population-based study. In 2011, self-reported health outcomes were compared using multivariable logistic regression across male veterans (n=53,406); active duty service members (n=2144); National Guard/Reserve service members (n=3724); and civilians (n=110,116). Multivariate logistic regression results are presented. Despite better healthcare access, veterans had poorer health and functioning than civilians and National Guard/Reserve members on several indicators. Veterans also were more likely than those on active duty to report diabetes. Veterans were more likely to report current smoking and heavy alcohol consumption than National Guard/Reserve members and civilian men, and lack of exercise compared to active duty men and National Guard/Reserve members. National Guard/Reserve men had higher levels of obesity, diabetes, and cardiovascular disease (versus active duty and veterans, active duty, and civilians, respectively). Active duty men were more likely to report current smoking and heavy alcohol consumption than civilians and National Guard/Reserve members, and reported more smokeless tobacco use than civilians. Veterans have poorer health and health behaviors; increased prevention efforts are needed from veteran-serving organizations. Despite good health, active duty men reported unhealthy lifestyles, indicating an important area for prevention efforts.


A BRCA1 mutant alters G2-M cell cycle control in human mammary epithelial cells.
Mutations in BRCA1 increase the risk of breast and ovarian cancer. Although the mechanism by which mutant BRCA1 alters growth regulation is unknown, the COOH terminus of BRCA1 appears to play a critical role. To examine this, we introduced a vector expressing BRCA1 COOH-terminal residues 1293-1863 (CT-BRCA1) into nontumorigenic human breast epithelial cells. Overexpression of CT-BRCA1 led to a reduction in the doubling time (from 64 to 44 h) and a decreased reliance on growth factors, suggesting that this CT-BRCA1 may function in a dominant-negative manner. Expression of CT-BRCA1 induced alterations in cell cycle control, mainly in G2-M, including a loss of G2-M block by colchicine. These results suggest that one function of BRCA1-related growth control occurs by governing checkpoint(s) between DNA replication and mitosis.


Mortality of employees of the United Kingdom Atomic Energy Authority, 1946-1979.
An analysis was conducted of 3373 deaths among 39 546 people employed by the United Kingdom Atomic Energy Authority between 1946 and 1979, the population having been followed up for an average of 16 years. Overall the death rates were below those prevailing in England and Wales but consistent with those expected in a normal workforce. At ages 15-74 years the standardised mortality ratios (SMRs) were 74 for deaths from all causes and 79 for deaths from all cancers. Mortality from only four causes was above the national average--namely, testicular cancer (SMR 153; 10 deaths), leukaemia (SMR 123; 35 deaths), thyroid cancer (SMR 122; three deaths), non-Hodgkin's lymphoma (SMR 107; 20 deaths)--but in none was the increase significant at the 5% level. Half of the authority's employees were recorded as having been monitored for exposure to radiation, their collective recorded exposure being 660 Sv (65 954 rem). Among these prostatic cancer was the only condition with a clearly increased mortality in relation to exposure. Of the 19 men who had a radiation record and died from prostatic cancer at ages 15-74 years, nine had been monitored for several different sources of exposure to radiation. The standardised mortality ratios were 889 (six deaths) in employees monitored for contamination by tritium, 254 (nine deaths) in those monitored for contamination by other radionuclides, and 385 (nine deaths) in those with dosimeter readings totalling more than 50 mSv (5 rem); but the same nine subjects tended to account for each of these significantly raised ratios. Because multiple exposures were common and other relevant information was not available the reason for the increased mortality from prostatic cancer in this population could not be determined and requires further investigation. Excess mortality rates of 2.2 and 12.5 deaths per million person years per 10 mSv (1 rem) were estimated for leukaemia and all cancers, respectively. The confidence limits around these estimates were wide, included zero, and made it unlikely that the International Commission on Radiological Protection's cancer risk coefficients were underestimated by more than 15-fold. Thus despite this being the largest British workforce whose mortality has been reported in relation to low level ionising radiation exposure, even larger populations will need to be followed up over longer periods before narrower ranges of risk estimates can be derived.


Iron and zinc bioavailabilities to pigs from red and white beans (Phaseolus vulgaris L.) are similar.
Common beans contain relatively high concentrations of iron (Fe) and zinc (Zn) but are also high in polyphenols and phytates, factors that may inhibit Fe and Zn absorption. In vitro (Caco-2 cells) and in vivo (pigs) models were used to compare Fe and Zn bioavailabilities between red and white beans, which differ in polyphenol content. Bean/maize diets containing 37% of either white or red cooked beans were formulated. Fe uptake by Caco-2 cells was 14-fold higher from the white bean diet compared to the red bean diet. The diets were fed to anemic piglets (n = 10) for 35 days. On experiment days 7 and 21, pigs were given meals containing beans intrinsically labeled with stable isotopes of Fe and Zn ((58)Fe, (70)Zn), followed by intravenous (iv) injections of (54)Fe and (67)Zn, to assess Fe and Zn absorption. Isotope ratios determined by inductively coupled plasma mass spectrometry in whole blood and plasma samples were used to calculate iron and zinc absorption, respectively. On day 35, animals were killed and duodenal sections were collected for DMT1 gene expression analysis. Fe absorption was 14 and 16% from the first labeled meal and 9 and 10.5% from the second labeled meal for the white and red beans, respectively (P > 0.05). Zn absorption was 28 and 23% from the first meal (P > 0.05) and 31 and 29% from the second meal (P > 0.05) for the white and red beans, respectively. DMT1 gene expression did not differ between treatments. It was concluded that bean color does not affect Fe or Zn bioavailability in vivo and that beans are a good source of bioavailable Fe and Zn.