Altered ETB- but not ETA-receptor density and function in sheep airway smooth muscle cells in culture.
The possibility that significant changes in endothelin (ET)A- and ETB-receptor density and function occur in airway smooth muscle cells (ASMCs) during cell growth and extended cell culture was investigated in sheep tracheal ASMCs. As in intact tracheal smooth muscle tissue from this species, early-passage sheep ASMCs contained a homogeneous population of ETA receptors. However, growth of ASMCs from seeding to postconfluence and repeated passage of ASMCs (6th to 14th passages) was associated with a substantial increase in ETB-receptor density, with no change in ETA-receptor density. ET-1-induced stimulation of ETB receptors increased the intracellular Ca2+ concentration in single ASMCs. Interestingly, a 2-day period of serum deprivation completely eliminated the increase in ETB-receptor density and the ETB receptor-mediated change in intracellular Ca2+ concentration. In summary, growth and repeated passage of sheep ASMCs were associated with a profound and selective increase in the density and function of the ETB receptor, a receptor subtype not present in early-passage ASMCs and not detected in intact sheep tracheal airway smooth muscle.


Coupling of insulin binding and insulin action on glucose transport in fat cells.
The initial lag phase of insulin action on glucose transport in adipocytes reflects an unknown process that couples receptor binding and glucose transport activity. The influence of temperature, cellular ATP, cyclic AMP, and calcium on this process and a possible relation to internalization of insulin were studied. The Arrhenius plot of the coupling shows a break in slope at 30 degrees C; the activation energy below 30 degrees C is 17.5 kcal/mol. Reduction of cellular ATP by 70% prolongs the coupling process; initial binding and final maximal response of the glucose transport remain unaffected. Further reduction of ATP (greater than 90%) before addition of insulin abolishes the coupling completely. Reduction of ATP at different time points after addition of insulin blocks further activation; however, the actual state of activity is preserved. Calcium depletion by EDTA prolongs the coupling and decreases the maximal response. Internalization of insulin as determined in chloroquine-treated cells begins later than transport activation and is in contrast to transport activation not observable at 15 degrees C. In conclusion, the coupling is not related to internalization; it is ATP-dependent, whereas the initial binding and the activated transport system are ATP-independent. Calcium but not cyclic AMP might be second messenger or cofactor in the coupling process.


State Variations in Infant Feeding Regulations for Child Care.
Early care and education (ECE) settings have become primary targets for policy change in recent years. In our 2008 study, we assessed state and regional variation in infant feeding regulations for ECE and compared them to national standards. We conducted the same regulatory review to assess change over time. Because all but 2 states have updated their regulations, we hypothesized that states would have made substantial improvements in the number of regulations supporting infant feeding in ECE. For this cross-sectional study, we reviewed infant feeding regulations for all US states for child care centers (centers) and family child care homes (homes). We compared regulations with 10 national standards and assessed the number of new regulations consistent with these standards since our previous review. Comparing results from 2008 and 2016, we observed significant improvements in 7 of the 10 standards for centers and 4 of the 10 standards for homes. Delaware was the only state with regulations meeting 9 of the 10 standards for centers in 2008. In 2016, Delaware and Michigan had regulations meeting 8 of the 10 standards. Previously, Arkansas, the District of Columbia, Minnesota, Mississippi, Ohio, and South Carolina had regulations consistent with 4 of the 10 standards for homes. In 2016, Delaware, Mississippi, and Vermont had regulations meeting 7 of the 10 standards. Evidence suggests that enacting new regulations may improve child health outcomes. Given that many states recently enacted regulations governing infant feeding, our findings point to the growing interest in this area.


Disinfectants and antiseptics: mechanisms of action and resistance.
Chemical biocides are used for the prevention and control of infection in health care, targeted home hygiene or controlling microbial contamination for various industrial processes including but not limited to food, water and petroleum. However, their use has substantially increased since the implementation of programmes to control outbreaks of methicillin-resistant Staphylococcus aureus, Clostridioides difficile and severe acute respiratory syndrome coronavirus 2. Biocides interact with multiple targets on the bacterial cells. The number of targets affected and the severity of damage will result in an irreversible bactericidal effect or a reversible bacteriostatic one. Most biocides primarily target the cytoplasmic membrane and enzymes, although the specific bactericidal mechanisms vary among different biocide chemistries. Inappropriate usage or low concentrations of a biocide may act as a stressor while not killing bacterial pathogens, potentially leading to antimicrobial resistance. Biocides can also promote the transfer of antimicrobial resistance genes. In this Review, we explore our current understanding of the mechanisms of action of biocides, the bacterial resistance mechanisms encompassing both intrinsic and acquired resistance and the influence of bacterial biofilms on resistance. We also consider the impact of bacteria that survive biocide exposure in environmental and clinical contexts.


Enhanced nitrogen removal with an onsite aerobic cyclic biological treatment unit.
Coastal Zone Act Reauthorization Amendments (CZARA, Section 6217) necessitate the requirement that onsite wastewater disposal units located near impaired surface waters or groundwater to provide at least 50% nitrogen removal. Approximately 38% of Hawaii households use onsite systems including septic tanks and cesspools that cannot meet this requirement. Upgrades to aerobic treatment units (ATUs) are a possible compliance solution. In Hawaii, ATUs must meet National Sanitation Foundation Standard 40 (NSF40) Class I effluent criteria. Previously, a multi-chamber, flow-through, combined attached/suspended growth type ATU (OESIS-750) and presently, a sequencing batch type ATU (CBT 0.8KF-210) were evaluated for NSF40 compliance, nutrient removal capability (NSF245), and adaptability for water reuse (NSF350). Both units easily achieved the NSF40 Class I effluent criteria. While the OESIS-750 achieved only 19% nitrogen removal, the CBT unit achieved 81% nitrogen removal, meeting the NSF245 criteria and CZARA requirements for applications in critical wastewater disposal areas. In addition, the CBT consistently produced effluent with turbidity less than 2 NTU (NSF350) and UVT254 greater than 70%, facilitating the production of unrestricted-use recycled water.


The saga of MHC-bound peptides: a renaissance for antigen presentation?
In this issue of the JCI, two separate studies on MHC-bound peptides reopen the debate on the utility of peptides for the purposes of vaccination and treatment of autoimmune diseases. In the first study, by Wahlström et al., peptides bound to HLA-DR17 from bronchoalveolar lavage cells of sarcoidosis patients were analyzed in order to identify target antigens of the autoimmune response (see the related article beginning on page 3576). In the second study, by Le Gall et al., the modulation of epitope immunodominance and the processing and presentation of HIV peptides for MHC class I recognition were shown to be dependent on flanking residues that were N terminal to the natural epitopes (see the related article beginning on page 3563). Both studies highlight the tremendous therapeutic potential of MHC-bound peptides. They also emphasize that technical issues are still plaguing this field and hindering our understanding of MHC presentation in vivo.


Recent Progress on Layered Cathode Materials for Nonaqueous Rechargeable Magnesium Batteries.
Rechargeable magnesium batteries (RMBs) are promising candidates for next-generation energy storage systems owing to their high safety and the low cost of magnesium resources. One of the main challenges for RMBs is to develop suitable high-performance cathode materials. Layered materials are one of the most promising cathode materials for RMBs due to their relatively high specific capacity and facile synthesis process. This review focuses on recent progress on layered cathode materials for RMBs, including layered oxides, sulfides, selenides, and other layered materials. In addition, effective strategies to improve the electrochemical performance of layered cathode materials are summarized. Moreover, future perspectives about the application of layered materials in RMBs are also discussed. This review provides some significant guidance for the further development of layered materials for RMBs.


Description of Karyotype of Kerodon acrobata, an endemic rodent in Brazilian Cerrado.
Kerodon acrobata is a caviidae rodent endemic from Brazilian Cerrado. It was described only in 1997 and the data about it is very scarce. The aim of this work was to characterize the karyotype of K. acrobata. Giemsa staining, nucleolar organizer region (NOR) banding, C-positive heterochromatin banding and DAPI fluorescence were used in N metaphases of a specimen collected in Asa Branca Farm, in Aurora do Tocantins municipality, Tocantins state, Brazil. K. acrobata showed the same diploid number, fundamental number and chromosome morphology as Kerodon rupestris. But its NOR location and heterochromatin distribution patterns indicated a unique cytogenetic profile when compared to its sister species, emphasizing the evolutionary uniqueness of this relatively new and unknown species. This record also extends the distribution of this species northward.


Chest physiotherapy: the effect on arterial oxygenation.
A prospective study of 47 patients during mechanical ventilation with positive end-expiratory pressure was undertaken to evaluate changes in Pao2 following chest physiotherapy. In contrast to other studies, we found no significant change in Pao2 following chest physiotherapy (p greater than 0.05). Different technics for physiotherapy and ventilation may have accounted for the absence of hypoxemia reported by others. Possible reasons for failure to improve Pao2 were suggested. Chest X-ray before and within 24 hours of physiotherapy showed improvement in 68 percent of patients. Chest physiotherapy was most effective in the treatment of unilobar densities and produced dramatic improvement in atelectasis of acute onset.


Intranasal Vaccination Affords Localization and Persistence of Antigen-Specific CD8⁺ T Lymphocytes in the Female Reproductive Tract.
Immunization strategies generating large numbers of antigen-specific T cells in the female reproductive tract (FRT) can provide barrier protection against sexually-transmitted pathogens, such as the human immunodeficiency virus (HIV) and human papillomaviruses (HPV). The kinetics and mechanisms of regulation of vaccine-induced adaptive T cell-mediated immune responses in FRT are less well defined. We present here evidence for intranasal delivery of the model antigen ovalbumin (OVA) along with alpha-galactosylceramide adjuvant as a protein vaccine to induce significantly higher levels of antigen-specific effector and memory CD8⁺ T cells in the FRT, relative to other systemic and mucosal tissues. Antibody blocking of the CXCR3 receptor significantly reduced antigen-specific CD8⁺ T cells subsequent to intranasal delivery of the protein vaccine suggesting an important role for the CXCR3 chemokine-receptor signaling for T cell trafficking. Further, intranasal vaccination with an adenoviral vector expressing OVA or HIV-1 envelope was as effective as intramuscular vaccination for generating OVA- or ENV-specific immunity in the FRT. These results support the application of the needle-free intranasal route as a practical approach to delivering protein as well as DNA/virus vector-based vaccines for efficient induction of effector and memory T cell immunity in the FRT.


Factors controlling selectivity in the ring-opening metathesis polymerization of 3-substituted cyclooctenes by monoaryloxide pyrrolide imido alkylidene (MAP) catalysts.
The origins of regio- and stereoselectivity in the ring-opening metathesis polymerization of 3-substituted cis-cyclooctenes by monoaryloxide pyrrolide imido alkylidene (MAP) Mo- and W-based catalysts are determined at the M06-2X/SDD|6-311+G(2df,p)//M06-L/SDD|6-31G(d)|MIDI! level of density functional theory. Considering cis-cyclooctene (COE) and 3-methyl-cis-cyclooctene (3MCOE) as monomers and W(N-t-Bu)(CH-t-Bu)(OHMT)(Pyr) (OHMT = hexamethylterphenoxide, Pyr = pyrrolide) as a catalyst, all possible syn and anti combinations of alkylidene and cyclic olefin, relative to the imido ligand, are evaluated. The observed Z-selectivity for the ring-opening metathesis (ROM) of COE is due to the large size of the aryloxide ligand, which forces both the alkylidene and the incoming cyclic olefin to be syn relative to the imido ligand. As determined previously for Grubbs' second-generation catalyst (G2), breakdown of the metallacyclobutane intermediate is the rate-limiting step for cyclic olefins having ring sizes exceeding five carbon atoms. Contrary to the G2 case, however, the ring-opening of 3MCOE by MAP catalysts prefers a proximal (3-substituent closest to the metal center) over a distal (3-substituent furthest from the metal center) approach. In all calculated paths, we observe inversion of catalyst configuration after each catalytic cycle.


[Comparison of GnRH antagonist fixed protocol and GnRH agonists long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer cycle].
To compare the clinical outcomes of gonadotropin-releasing hormone (GnRH) antagonist (GnRH-ant) fixed protocol with GnRH agonist (GnRH-a) long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer (IVF-ET) cycle, and to explore the feasibility and advantage of GnRH antagonist protocol performed in normal responders. From January 2011 to June 2011, 771 infertile women with normal ovarian reserve function underwent their first IVF or intracytoplasmic sperm injection (ICSI) cycles in Peking University Third Hospital, which were divided into 245 cycles in GnRH-ant fixed protocol group (GnRH-ant group) and 526 cycles in GnRH-a long protocol group (GnRH-a group). The data of general demographic, treatment and clinical outcome were compared between two groups. Age, infertile duration, body mass index (BMI), baseline serum follicle-stimulating hormone (FSH) and estradiol levels between two groups did not reached statistical difference (P > 0.05). The level of estradiol was (12 289 ± 6856) pmol/L in GnRH-ant group and (14 934 ± 8007) pmol/L in GnRH-a group at day of hCG injection. The mean length of stimulation was (10.3 ± 1.2) days in GnRH-ant group and (12.8 ± 1.6) days in GnRH-a group. The dose of gonadotropin was (2013 ± 607) U in GnRH-ant group and (2646 ± 913) U in GnRH-a group. The number of ovum was 15 ± 7 in GnRH-ant group and 17 ± 8 in GnRh-a group. Those clinical parameter all reached statistical difference (P < 0.05). The number of embryo was 7 ± 4 in GnRH-ant group and 8 ± 5 in GnRH-a group, the rate of clinical pregnancy was 40.9% (94/230) in GnRH-ant group and 45.6% (216/474) in GnRH-a group, the rate of implantation was 26.1% (128/490) in GnRH-ant group and 30.9% (307/994) in GnRH-a group, the rate of continuing pregnancy was 38.7% (89/230) in GnRH-ant group and 42.6% (202/474)in GnRH-a group, those parameter did not reach statistical difference (P > 0.05). The rate of moderate or severe ovarian hyperstimulation syndrome was 2.4% (6/245) in GnRH-ant group and 4.2% (22/526) in GnRH-a group, which did not show significant difference (P > 0.05). In the first IVF or ICSI cycle of the patients with normal ovarian reserve function, the fixed GnRH-ant protocol could get the same satisfied clinical outcome, and it is more economic, convenient and safer compared with low dose depot GnRH-a long protocol.


Visible-light assisted methylene blue (MB) removal by novel TiO(2)/adsorbent nanocomposites.
In present work, visible light sensitive TiO(2)/adsorbent nanocomposites (TNC) were prepared via a facile wet chemical method. Three types of adsorbents including zeolites (F-9, HSZ-690 and HSZ-930), mesoporous silica (MPS-2.7 and 4) and activated carbon, were used as the porous substrates for nanocomposites. Visible light sensitivity was incorporated to TNCs by nitrogen doping of TiO(2), which is obtained through the addition of a nitrogen precursor, triethylamine, within the same wet chemical procedure. The photocatalytic and adsorption ability of as-prepared TNC resultants were studied using solutions of methylene blue (MB) as a model pollutant. Synergistic interactions between adsorption and visible-light photocatalysis were observed, as under the assistance of visible-light irradiation all TNCs achieved higher MB removal rates than those by adsorption process alone. The better performance of the as-prepared N-doped TNC reveals its potential to be used for cost effective solar photocatalytic degradation of dissolved organic compounds.


Dosage compensation of the copia retrotransposon in Drosophila melanogaster.
Dosage compensation in Drosophila has been studied at the steady state RNA level for several single-copy genes; however, an important point is addressed by analyzing a repetitive, transposable element for dosage compensation. The two issues of gene-specific cis control and genomic position can be studied by determining the extent of dosage compensation of a transposable element at different chromosomal locations. To determine whether the multicopy copia transposable element can dosage compensate, we used the X-linked white-apricot (wa) mutation in which a copia element is present. The extent of dosage compensation was determined for the white and copia promoters in larvae and adults in two different genomic locations of the wa allele. We conclude that copia is able to dosage compensate, and that the white promoter and the copia promoter are not coordinate in their dosage compensation abilities when assayed under these various conditions. Thus, two transcriptional units, one within the other, both of which are able to dosage compensate, do so differently in response to developmental stage and genomic position.


Dendritic cell-based vaccines in breast and gynaecologic cancer.
Major advances in understanding the functional interactions between tumour cells and the host immune system, in particular the generation and regulation of T cell immunity, have revived interest in cancer vaccination strategies. A crucial step for mounting an anti-tumour response is the capture, processing and presentation of tumour antigens (TA) to cognate T cells by professional antigen-presenting cells (APC), followed by their activation and clonal proliferation. Dendritic cells (DC) are potent APC with the unique ability to stimulate primary immune responses. Animal models have demonstrated that TA-charged DC can activate specific cytotoxic T cells (CTL) and even regression of established tumours in cancer-bearing hosts. These findings, as well as the elaboration of methods for generating large numbers of DC ex vivo, have provided a compelling rationale for using DC as potent adjuvants to deliver TA to the immune system in order to trigger or amplify an inadequate response. The capacity of TA-pulsed DC to induce significant CTL immunity translating into occasional therapeutic benefit has been documented in several clinical settings including B cell lymphoma, myeloma, melanoma, prostate, colon, ovarian and renal cell carcinoma. In this review, we summarize key biological functions of DC and focus on recent DC-based vaccination trials of breast, ovarian and cervical cancer.


Beyond quantitative proteomics: signal enhancement of the a1 ion as a mass tag for peptide sequencing using dimethyl labeling.
Stable isotope-based dimethyl labeling that produces a dimethyl labeled terminal amine or a monomethylated proline N-terminus by reductive methylation (Anal. Chem. 2003, 75, 6843-6852) was reported as a promising strategy for global quantitative proteomics because of the simplicity of the process and its fast and complete reaction. This labeling strategy provides a signal enhancement for the produced a1 ions, which are usually hard to detect among most of the nonderivatized fragments. To assist peptide sequencing, in this study, the enhanced a1 ion produced under either collision induced dissociation (CID) or post source decay (PSD) modes was further characterized and applied as a mass tag for fingerprinting the identity of N-terminal amino acid. On the basis of the analysis of standard peptides, tryptic digests of hemoglobin and cell lysates, it was proved that such signal enhancement occurred to a1 ions derived from all 20 of the amino acids residues and this phenomenon was explained based the formation of stable quaternary immoniun ions. Accurate determination of a1 ions was shown to increase the chance for peptide de novo sequencing and also provided higher confidence in the scores obtained when identifying a protein through database searching. In addition, the a1 ion was further demonstrated to be used as a universal tag for precursor ion scan in a Q-TOF instrument, leading to a greater number of peptide ions sequenced. Combined with the capability for differential quantitation, the stable isotope-based dimethyl labeling increases the usefulness of the labeling method for MS-based proteomics.


Evaluation of Density Functionals, SCC-DFTB, Neglect of Diatomic Differential Overlap (NDDO) Models and Molecular Mechanics Methods for Prolyl-Leucyl-Glycinamide (PLG) and Structural Derivatives.
Prolyl-leucyl-glycinamide (PLG) is a unique endogenous peptide that modulates dopamine receptor subtypes of the D(2) receptor family within the CNS. We seek to elucidate the structural basis and molecular mechanism by which PLG and its analogues modulate dopamine receptors, toward the development of new therapeutics to treat Parkinson's disease, tardive dyskinesia and schizophrenia. As a first step toward establishing a validated protocol for accurate computational modeling of PLG and associated peptidomimetic analogues, we evaluated the accuracy of density functional theory (DFT), wavefunction theory (WFT), and molecular mechanics (MM) calculations for PLG and for a library of structurally related small molecules. We first tested twelve local and nonlocal density functionals, Hartree-Fock (HF) theory, four "semiempirical" methods of the neglect of diatomic differential overlap (NDDO) type, and one self-consistent-charge nonorthogonal tight-binding (SCC-DFTB) method as implemented in two software suites, against coupled-cluster benchmark geometries for 4-methylthiazolidine, a small molecule that comprises key structural features present in our PLG analogue library. DFT and HF calculations were done with the MG3S augmented polarized triple-zeta basis set. We find that for 4-methylthiazolidine bond distances, DFT significantly outperforms NDDO, and both SCC-DFTB versions we evaluated perform worse than HF theory and are less accurate than 83% of the density functionals tested. The top five functionals for 4-methylthiazolidine were M05-2X, mPW1PW, B97-2, M06-2X, and PBEh, with mean unsigned errors (MUEs) in bond length of 0.0017, 0.0020, 0.0023, 0.0025 and 0.0027 Å, respectively. The widely used B3LYP functional ranked 11(th) out of twelve functionals evaluated, slightly below SCC-DFTB, and is significantly less accurate for 4-methylthiazolidine bond distances (MUE = 0.0095 Å) than the best local functional (M06-L, MUE = 0.0030 Å), which is far less computationally costly. Based on that initial analysis, we obtained new M05-2X benchmark geometric parameters for PLG and a library of eleven peptidomimetic derivatives, which we in turn used to examine the accuracy of thirty-four popular molecular mechanics (MM) force fields, four NDDO approaches, and SCC-DFTB for the full compound structures. Here, we found that ∼70% of the MM force fields tested superior to the best semiempirical and SCC-DFTB codings. Moreover, AMBER-type force fields proved most accurate among MM methods for this class of small-molecule peptidomimetics; the AMBER-type methods comprised eight out of the top ten molecular mechanics options we tested.


Barbiturate receptor sites are coupled to benzodiazepine receptors.
Barbiturates enhance the binding of [3H]diazepam to benzodiazepine receptor sites in rat brain. This effect occurs at pharmacologically relevant concentrations of barbiturates, and the relative activity of a series of compounds correlates highly with anesthetic activity of the barbiturates and with their ability to enhance postsynaptic inhibitory responses to the neurotransmitter gamma-aminobutyric acid. Barbiturate enhancement of benzodiazepine binding is stereospecific, with the more active anesthetic isomers of N1-methylbarbiturates being also more active than their stereoisomers in enhancing benzodiazepine binding. The active barbiturates produce a reversible enhancement in the affinity of specific benzodiazepine binding with no effect on the number of binding sites. The barbiturate enhancement, but not the baseline benzodiazepine binding, is competitively inhibited by the convulsant picrotoxinin (at 1-10 microM), a drug that has been shown to label barbiturate-sensitive brain membrane sites related to the gamma-aminobutyric acid receptor-ionophore complex. The barbiturate effect is also dependent upon the presence of certain anions, and only those anions, that penetrate the chloride channels regulated by gamma-aminobutyric acid receptors. These results suggest that picrotoxin-sensitive barbiturate binding sites are coupled to benzodiazepine receptors in the gamma-aminobutyric acid receptor-ionophore complex, and that these binding sites have the properties of pharmacologically relevant receptors that mediate at least part of the action of various nervous system depressant and excitatory drugs.


[The role of cyclic nucleotides in the processes of adaptation and alteration of body radiosensitivity].
The experimental data on the role of the cyclic nucleotides (cAMP, cGMP) in the mammalian integrating systems and their significance in the processes of adaptation and radiosensitivity change of animals are analysed. The changes in the system of cyclic nucleotides (CN) are reported for a number of morbid conditions at the subunit level. The CN system changes as a body response to the extreme action including ionizing radiation are revealed. A concept on a phase correlation between adrenergic and cholinergic system activity in development of adaptive reaction of a cell and a body on the whole during radiation or the action of some other factors is emphasized. The significance of cAMP/cGMP ratio calculation (coefficient K) is underlined for the analysis and prediction of the morbid state outcome.


A Conditional Normalizing Flow for Accelerated Multi-Coil MR Imaging.
Accelerated magnetic resonance (MR) imaging attempts to reduce acquisition time by collecting data below the Nyquist rate. As an ill-posed inverse problem, many plausible solutions exist, yet the majority of deep learning approaches generate only a single solution. We instead focus on sampling from the posterior distribution, which provides more comprehensive information for downstream inference tasks. To do this, we design a novel conditional normalizing flow (CNF) that infers the signal component in the measurement operator's nullspace, which is later combined with measured data to form complete images. Using fastMRI brain and knee data, we demonstrate fast inference and accuracy that surpasses recent posterior sampling techniques for MRI. Code is available at https://github.com/jwen307/mri_cnf.


Update on new muscle glycogenosis.
The field of muscle glycogenoses has progressed in recent years by the identification of new disorders, and by reaching a better understanding of pathophysiology of the disorders and the physiology of glycogen metabolism. In this review, we describe the clinical and pathological features of the three most recently described muscle glycogenoses caused by recessive mutations in GYG1, RBCK1 and PGM1. The three involved enzymes play different roles in glycogen metabolism. Glycogenin-1 (GYG1) is involved in the initial steps of glycogen synthesis, whereas phosphoglucomutase catalyzes two metabolic pathways; the connection between galactose and glycogen on one side, and glucose metabolism on the other side. The metabolic consequences of mutations in the ubiquitin ligase gene RBCK1 are still poorly understood. GYG1 deficiency has been associated with cardiomyopathies with abnormal storage material in the heart, but most cases present with a polyglucosan body myopathy without cardiac involvement. The recent identification of new glycogenosis not only allows to improve the knowledge of glycogen metabolism, but also builds bridges with protein glycosylation and immune system.


Use of the BioGRID Database for Analysis of Yeast Protein and Genetic Interactions.
The BioGRID database is an extensive repository of curated genetic and protein interactions for the budding yeast Saccharomyces cerevisiae, the fission yeast Schizosaccharomyces pombe, and the yeast Candida albicans SC5314, as well as for several other model organisms and humans. This protocol describes how to use the BioGRID website to query genetic or protein interactions for any gene of interest, how to visualize the associated interactions using an embedded interactive network viewer, and how to download data files for either selected interactions or the entire BioGRID interaction data set.


Triton's Global Heat Budget.
Internal heat flow from radioactive decay in Triton's interior along with absorbed thermal energy from Neptune total 5 to 20 percent of the insolation absorbed by Triton, thus comprising a significant fraction of Triton's surface energy balance. These additional energy inputs can raise Triton's surface temperature between approximately 0.5 and 1.5 K above that possible with absorbed sunlight alone, resulting in an increase of about a factor of approximately 1.5 to 2.5 in Triton's basal atmospheric pressure. If Triton's internal heat flow is concentrated in some areas, as is likely, local effects such as enhanced sublimation with subsequent modification of albedo could be quite large. Furthermore, indications of recent global albedo change on Triton suggest that Triton's surface temperature and pressure may not now be in steady state, further suggesting that atmospheric pressure on Triton was as much as ten times higher in the recent past.


On the binding of tumor necrosis factor (TNF) to heparin and the release in vivo of the TNF-binding protein I by heparin.
Tumor necrosis factor (TNF), a protein released by activated macrophages, is a central mediator of the host response to infection and inflammation. The TNF-binding protein I (TNF-BP-I) is a soluble fragment of the p60 transmembrane TNF receptor and an antagonist to TNF. The level of serum TNF-BP-I was found to be increased in patients with renal insufficiency as a result of a decrease in the glomerular filtration rate. During hemodialysis of patients with renal failure there was a rapid but transient increase in serum TNF-BP-I. This increase was found to be caused by heparin given before dialysis and a similar dose-dependent response to heparin was observed also in healthy individuals. The finding of a repeated release of TNF-BP-I into the circulation with intermittent injections of heparin indicates that TNF-BP-I is present both in a storage pool and in a circulating pool. The mechanism for the heparin-mediated release of TNF-BP-I was not explained; TNF-BP did not show affinity for heparin. On the other hand, TNF was found to have affinity for heparin and it could also be dissociated from heparin by TNF-BP-I. It is suggested that heparin-like molecules of the extracellular matrix can retain TNF in physical proximity with target cells and restrict the actions of TNF and protect against systemic harmful manifestations.


The association between electronic health information usage and patient-centered communication: a cross sectional analysis from the Health Information National Trends Survey (HINTS).
Patient-provider communication can be assessed by the patient-centered communication (PCC) score. With rapid development of electronic health (eHealth) information usage, we are uncertain of their role in PCC. Our study aims to determine the association between PCC and eHealth usage with the analysis of national representative survey data. This is a cross sectional analysis using the Health Information National Trends Survey 5 (HINTS 5) cycle 1 to cycle 4 data (2017-2020). Seven specific questions were used for PCC assessment, and eHealth usage was divided into two types (private-eHealth and public-eHealth usage). A multivariate logistic regression was performed to determine the association between PCC and eHealth usage after the adjustment of other social, demographic, and clinical variables. Our study analyzed a total of 13,055 unweighted participants representing a weighted population of 791,877,728. Approximately 43% of individuals used private eHealth and 19% used public eHealth. The adjusted odds ratio (AOR) of private-eHealth usage associated with positive PCC was 1.17 (95% CI 1.02-1.35, p = 0.027). The AOR of public-eHealth usage associated with positive PCC was 0.84 (95% CI 0.71-0.99, p = 0.043). Our study found that eHealth usage association with PCC varies. Private-eHealth usage was positively associated with PCC, whereas public-eHealth usage was negatively associated with PCC.


Immunologic potency of recombined A- and B-chains of synthetic human and pancreatic pork insulins.
Pork insulin recombined from cleaved A- and B-chains of pancreatic insulin was shown to be identical with the pancreatic hormone both in erythrocyte receptor assays and in immunoassays employing four different antisera, including two species-specific human anti-insulin sera. Pancreatic human insulin and that prepared from recombined bacterially synthesized A- and B-chains were also indistinguishable in the same systems. This study demonstrates that the prohormone stage of either human or porcine insulin is not required to preserve potency of membrane binding or immunochemical reactivity, even employing an antiserum with marked species specificity.


[Toxicity and proteolysis in gnotobiotic rats with thermal burns].
Experiments were conducted on germ-free (gnotobiotic) rats. A study was made of the toxic properties of the organs and the activity of proteolytic enzymes in them after thermal burns. Despite the absence of pathogenic microbial flora the germ-free rats developed the state of toxemia with an increase of the level of proteolytic enzymes. The extent of the toxic and catabolic reaction depended on the severity of the burn. Intoxication and an increase in proteolysis in the gnotobiotic rats were similar, and, in a number of cases greater than in usual animals. A conclusion was drawn on the important role of the tissue sources of intoxication in burns. A correlation between the toxemia and proteolysis was established.


In silico Docking Analysis for Blocking JUNO-IZUMO1 Interaction Identifies Two Small Molecules that Block in vitro Fertilization.
Combined hormone drugs are the basis for orally administered contraception. However, they are associated with severe side effects that are even more impactful for women in developing countries, where resources are limited. The risk of side effects may be reduced by non-hormonal small molecules which specifically target proteins involved in fertilization. In this study, we present a virtual docking experiment directed to discover molecules that target the crucial fertilization interactions of JUNO (oocyte) and IZUMO1 (sperm). We docked 913,000 molecules to two crystal structures of JUNO and ranked them on the basis of energy-related criteria. Of the 32 tested candidates, two molecules (i.e., Z786028994 and Z1290281203) demonstrated fertilization inhibitory effect in both an in vitro fertilization (IVF) assay in mice and an in vitro penetration of human sperm into hamster oocytes. Despite this clear effect on fertilization, these two molecules did not show JUNO-IZUMO1 interaction blocking activity as assessed by AVidity-based EXtracellular Interaction Screening (AVEXIS). Therefore, further research is required to determine the mechanism of action of these two fertilization inhibitors.


EMIRGE: reconstruction of full-length ribosomal genes from microbial community short read sequencing data.
Recovery of ribosomal small subunit genes by assembly of short read community DNA sequence data generally fails, making taxonomic characterization difficult. Here, we solve this problem with a novel iterative method, based on the expectation maximization algorithm, that reconstructs full-length small subunit gene sequences and provides estimates of relative taxon abundances. We apply the method to natural and simulated microbial communities, and correctly recover community structure from known and previously unreported rRNA gene sequences. An implementation of the method is freely available at https://github.com/csmiller/EMIRGE.


Ambulatory diagnosis of sleep-disordered breathing: Pushing out the boundaries.
Advances in signal technology facilitate the ambulatory diagnosis of obstructive sleep apnea and represent an important development in the management of this highly prevalent disorder. The recent report of Traverso and co-authors describes a novel diagnostic approach by an ingestible vital-monitoring pill that is capable of detecting sleep apnea.


Mitochondria in the flight muscles of insects. I. Chemical composition and enzymatic content.
1. The "indirect" thoracic muscles of adult dipterous and hymenopterous insects consist of a unique type of muscle characterized by the presence of numerous spherical, intracytoplasmic bodies termed "sarcosomes." 2. When the muscle is teased or ground, the sarcosomes are liberated as a turbid suspension of bodies ranging from 1 to 4 micro in diameter. A method is described for the isolation of sarcosomes by a simple differential centrifugation. 3. The cytochemical, chemical, and enzymatic properties of sarcosomes were examined for the purpose of appraising their relation to the cytoplasmic bodies of other tissues. 4. Fresh sarcosomes are slowly but selectively stained by the mitochondrial reagents, Janus green B and pinacyanol. Fixed sarcosomes give a positive reaction with Regaud's mitochondrial stain. 5. Chemical analyses show that approximately 29 per cent of the dry weight of sarcosomes consists of lipids and 60 per cent of protein. Microbiological assay indicates the presence of about 1 gamma of riboflavin per milligram of nitrogen. These values resemble those reported for isolated mitochondria of vertebrate liver and kidney. 6. When examined spectroscopically the sarcosomes, like the vertebrate mitochondria, show a high titer of cytochromes a, b, and c. 7. The titer of cytochrome oxidase varies systematically with the adult age of the insect. A similar relation is observed for the enzyme catalase. 8. Isolated sarcosomes show significant titers of succinoxidase, alpha-glycerophosphate dehydrogenase, malic dehydrogenase, and pyruvic dehydrogenase. The following dehydrogenases could not be demonstrated: xanthine, phenylalanine, glycine, lactic, choline, glutamic, and alcohol. These results are compared with those previously reported for vertebrate mitochondria. 9. In view of their manifold points of biochemical similarity, it is concluded that the sarcosomes are the mitochondria of this highly specialized muscular tissue.


Revascularization for patients with diabetes mellitus and stable ischemic heart disease: an update.
To provide an update on the management of patients with diabetes mellitus and requiring coronary revascularization. Evidence continues to show that patients with diabetes mellitus and ischemic heart disease represent a very high-risk group of patients. Choice of stent appears important for minimizing target lesion and target vessel adverse events with everolimus eluting stents having the best performance, particularly in patients being treated with insulin. The higher risk of adverse angioplasty results in patients with diabetes appears most related to the disease state per se and not necessarily to anatomical complexities. Interestingly, physiologic documentation of nonischemia producing lesions with use of fractional flow reserve appears less reassuring in this setting of aggressive and rapid atherosclerosis progression, particularly if myocardial infarction has occurred previously, than in patients without diabetes. Coronary artery bypass surgery in patients with appropriate anatomy and diabetes continues to emerge in many analyzes as the optimal, long-term therapy. The treatment of diabetes per se, advances in stent technology and optimization of coronary artery bypass techniques are all occurring in parallel making it very critical for the design of modern era trials that keep pace with these advances. Currently, in patients with appropriate anatomy who are willing candidates, bypass surgery remains the optimal, long-term therapeutic option.


Licochalcone A, a natural chalconoid isolated from Glycyrrhiza inflata root, induces apoptosis via Sp1 and Sp1 regulatory proteins in oral squamous cell carcinoma.
Licochalcone A (LCA), a chalconoid derived from root of Glycyrrhiza inflata, has been known to possess a wide range of biological functions such as antitumor, anti-angiogenesis, antiparasitic, anti-oxidant, antibacterial and anti-inflammatory effects. However, the anticancer effects of LCA on oral squamous cell carcinoma (OSCC) have not been reported. Our data showed that LCA inhibited OSCC cell (HN22 and HSC4) growth in a concentration- and time-dependent manner. Mechanistically, it was mediated via downregulation of specificity protein 1 (Sp1) expression and subsequent regulation of Sp1 downstream proteins such as p27, p21, cyclin D1, Mcl-1 and survivin. Here, we found that LCA caused apoptotic cell death in HSC4 and HN22 cells, as characterized by sub-G1 population, nuclear condensation, Annexin V staining, and multi-caspase activity and apoptotic regulatory proteins such as Bax, Bid, Bcl(-xl), caspase-3 and PARP. Consequently, this study strongly suggests that LCA induces apoptotic cell death of OSCC cells via downregulation of Sp1 expression, prompting its potential use for the treatment of human OSCC.


The Magic Touch: A Case Report of How Smartphone Fingerprint Technology Was Utilized to Establish a Last Known Well Time for Recanalization Treatment in a Patient With Acute Ischemic Stroke.
Tissue plasminogen activator (tPA) is currently a standard of care for acute stroke patients. One of the necessary criteria in determining eligibility for tPA is the last known well (LKW) time. The LKW time is unfortunately often difficult to obtain accurately if no witness is available, thus posing as an obstacle for acute recanalization therapy. We present the case of a patient who arrived unresponsive with an unwitnessed onset of symptoms concerning for an acute stroke. An LKW time was able to be successfully established by using her fingerprint to unlock her phone and discover a coherent text sent a few hours prior. Patient was able to receive intravenous (IV) tPA and demonstrated remarkable recovery. The use of fingerprint ID to unlock the patient's phone raises the concern of breach of privacy and whether involuntary smartphone searches apply to the emergency code of conduct outlined by the FDA. Smartphone applications, such as Apple iOS "Medical ID" argues for maximal utilization of smartphone technology for emergent medical conditions. Utilization of smartphone technology can potentially serve a potential solution, but the question remains as to whether this practice would be deemed to be ethically appropriate under the policy of implied informed consent under emergent conditions.


Intracellular recordings from renin-positive cells of the afferent glomerular arteriole.
Intracellular recordings were made in juxtaglomerular granulated (JG) cells and in vascular smooth muscle (VSM) cells in afferent arterioles of hydronephrotic mouse kidneys. Both cell types did not differ in their passive and active electrical membrane properties; membrane potential was about -58 mV, input resistance exceeded 400 M omega, and JG as well as VSM cells showed spontaneous depolarizations resembling excitatory junction potentials and active responses observed in smooth muscle cells of other blood vessels in various species. These depolarizations, attributed to spontaneous transmitter release from adrenergic terminals, were extremely polymorphous and quite frequent. Epinephrine, norepinephrine, phenylephrine, arginine vasopressin, and angiotensin II depolarized JG and VSM cells, but isoproterenol and orciprenaline had no effect. A hyperpolarizing action of catecholamines was never observed. It is suggested that, in this in vitro preparation, isoproterenol increases renin secretion by a mechanism independent of membrane potential changes. Depolarizations mediated by alpha-mimetic agents, arginine vasopressin, and angiotensin II, as well as by the junctional activity may inhibit renin secretion by an increased calcium influx into JG cells.


Effect of cocoa on blood pressure.
High blood pressure is an important risk factor for cardiovascular disease, contributing to about 50% of cardiovascular events worldwide and 37% of cardiovascular-related deaths in Western populations. Epidemiological studies suggest that cocoa-rich products reduce the risk of cardiovascular disease. Flavanols found in cocoa have been shown to increase the formation of endothelial nitric oxide which promotes vasodilation and therefore blood pressure reduction. Here we update previous meta-analyses on the effect of cocoa on blood pressure. To assess the effects on blood pressure of chocolate or cocoa products versus low-flavanol products or placebo in adults with or without hypertension when consumed for two weeks or longer. This is an updated version of the review initially published in 2012. In this updated version, we searched the following electronic databases from inception to November 2016: Cochrane Hypertension Group Specialised Register, CENTRAL, MEDLINE and Embase. We also searched international trial registries, and the reference lists of review articles and included trials. Randomised controlled trials (RCTs) investigating the effects of chocolate or cocoa products on systolic and diastolic blood pressure in adults for a minimum of two weeks duration. Two review authors independently extracted data and assessed the risks of bias in each trial. We conducted random-effects meta-analyses on the included studies using Review Manager 5. We explored heterogeneity with subgroup analyses by baseline blood pressure, flavanol content of control group, blinding, age and duration. Sensitivity analyses explored the influence of unusual study design. Thirty-five trials (including 40 treatment comparisons) met the inclusion criteria. Of these, we added 17 trials (20 treatment comparisons) to the 18 trials (20 treatment comparisons) in the previous version of this updated review.Trials provided participants with 30 to 1218 mg of flavanols (mean = 670 mg) in 1.4 to 105 grams of cocoa products per day in the active intervention group. The control group received either a flavanol-free product (n = 26 treatment comparisons) or a low-flavanol-containing cocoa powder (range 6.4 to 88 mg flavanols (mean = 55 mg, 13 treatment comparisons; 259 mg, 1 trial).Meta-analyses of the 40 treatment comparisons involving 1804 mainly healthy participants revealed a small but statistically significant blood pressure-reducing effect of flavanol-rich cocoa products compared with control in trials of two to 18 weeks duration (mean nine weeks):Mean difference systolic blood pressure (SBP) (95% confidence interval (CI): -1.76 (-3.09 to -0.43) mmHg, P = 0.009, n = 40 treatment comparisons, 1804 participants;Mean difference diastolic blood pressure (DBP) (95% CI): -1.76 (-2.57 to -0.94) mmHg, P < 0.001, n = 39 treatment comparisons, 1772 participants.Baseline blood pressure may play a role in the effect of cocoa on blood pressure. While systolic blood pressure was reduced significantly by 4 mmHg in hypertensive people (n = 9 treatment comparisons, 401 participants), and tended to be lowered in prehypertensive people (n= 8 treatment comparisons, 340 participants), there was no significant difference in normotensive people (n = 23 treatment comparisons, 1063 participants); however, the test for subgroup differences was of borderline significance (P = 0.08; I2 = 60%), requiring further research to confirm the findings.Subgroup meta-analysis by blinding suggested a trend towards greater blood pressure reduction in unblinded trials compared to double-blinded trials, albeit statistically not significant. Further research is needed to confirm whether participant expectation may influence blood pressure results. Subgroup analysis by type of control (flavanol-free versus low-flavanol control) did not reveal a significant difference.Whether the age of participants plays a role in the effect of cocoa on blood pressure, with younger participants responding with greater blood pressure reduction, needs to be further investigated.Sensitivity analysis excluding trials with authors employed by trials sponsoring industry (33 trials, 1482 participants) revealed a small reduction in effect size, indicating some reporting bias.Due to the remaining heterogeneity, which we could not explain in terms of blinding, flavanol content of the control groups, age of participants, or study duration, we downgraded the quality of the evidence from high to moderate.Results of subgroup analyses should be interpreted with caution and need to be confirmed or refuted in trials using direct randomised comparisons.Generally, cocoa products were highly tolerable, with adverse effects including gastrointestinal complaints and nausea being reported by 1% of participants in the active cocoa intervention group and 0.4% of participants in the control groups (moderate-quality evidence). This review provides moderate-quality evidence that flavanol-rich chocolate and cocoa products cause a small (2 mmHg) blood pressure-lowering effect in mainly healthy adults in the short term.These findings are limited by the heterogeneity between trials, which could not be explained by prespecified subgroup analyses, including blinding, flavanol content of the control groups, age of participants, or study duration. However, baseline blood pressure may play a role in the effect of cocoa on blood pressure; subgroup analysis of trials with (pre)hypertensive participants revealed a greater blood pressure-reducing effect of cocoa compared to normotensive participants with borderline significance.Long-term trials investigating the effect of cocoa on clinical outcomes are also needed to assess whether cocoa has an effect on cardiovascular events and to assess potential adverse effects associated with chronic ingestion of cocoa products.


Immunohistochemical localization of enkephalin in rat brain and spinal cord.
The distribution of immunoreactive enkephalin in rat brain and spinal cord was studied by immunoperoxidase staining using antiserum to leucine-enkephalin ([Leu5]-enkephalin) or methionine-enkephalin ([Met5]-enkephalin). Immunoreactive staining for both enkephalins was similarly observed in nerve fibers, terminals and cell bodies in many regions of the central nervous system. Staining of perikarya was detected in hypophysectomized rats or colchicine pretreated rats. The regions of localization for enkephalin fibers and terminals include in the forebrain: lateral septum, central nucleus of the amygdala, area CA2 of the hippocampus, certain regions of the cortex, corpus striatum, bed nucleus of the stria terminalis, hypothalamus including median eminence, thalamus and subthalamus; in the midbrain: nucleus interpeduncularis, periaqueductal gray and reticular formation; in the hind brain: nucleus parabrachialis, locus ceruleus, nuclei raphes, nucleus cochlearis, nucleus tractus solitarii, nucleus spinalis nervi trigemini, motor nuclei of certain cranial nerves, nucleus commissuralis and formatio reticularis; and in the spinal cord the substantia gelatinosa. In contrast enkephalin cell bodies appear sparsely distributed in the telencephalon, diencephalon, mesencephalon and rhombencephalon. The results of the histochemical staining show that certain structures which positively stain for enkephalin closely correspond to the distribution of opiate receptors in the brain and thus support the concept that the endogenous opiate peptides are involved in the perception of pain and analgesia. The localization of enkephalin in the preoptic-hypothalamic region together with the presence of enkephalin perikarya in the paraventricular and supraoptic nuclei suggest a role of enkephalin in the regulation of neuroendocrine functions.


Agglutinin from Alfalfa Necessary for Binding and Nodulation by Rhizobium meliloti.
A protein that specifically agglutinates Rhizobium meliloti, the alfalfa root nodule endosymbiont, has been purified from alfalfa seed. Material cross-reactive to antiserum prepared against the purified agglutinin is present in all alfalfa varieties that were tested but is absent in corn and other legumes not nodulated by Rhizobium meliloti. Studies with nonnodulating mutants of this microorganism incapable of binding to alfalfa roots suggest that the agglutinin is responsible for specific recognition between Rhizobium meliloti and alfalfa and that this recognition is an essential step in nodule formation.


High-Stringency Evaluation of the Automated BD Phoenix CPO Detect and Rapidec Carba NP Tests for Detection and Classification of Carbapenemases.
There is an urgent need for rapid, accurate detection and classification of carbapenemases. The current study evaluated the automated BD Phoenix CPO Detect and the manual bioMérieux Rapidec Carba NP tests for meeting these needs. Both tests were challenged with 294 isolates of Enterobacteriaceae spp., Pseudomonas aeruginosa, and Acinetobacter baumannii chosen to provide extreme diagnostic difficulty. Carbapenemases such as KPC, NMC-A, IMI, SME, NDM, SPM, IMP, VIM, and OXA-23, 40, 48, 58, 72, 181, and 232 were produced by 243 isolates and 51 carbapenemase-negative isolates included porin mutants and producers of extended-spectrum β-lactamases (ESBLs), AmpCs, K1, and broad-spectrum β-lactamases. Both tests exhibited high sensitivity of carbapenemase detection (>97%). Due to the highly challenging carbapenemase-negative isolates, specificities were lower than typical for evaluations involving mostly routine clinical isolates. BD Phoenix CPO Detect was 68.6% specific and Rapidec Carba NP was 60.8% to 78.4% specific, depending on how borderline results were interpreted. Only BD Phoenix CPO Detect classified carbapenemases. It correctly classified 85.0% of class A, 72.4% of class B, and 88.6% of class D carbapenemases. Importantly with respect to empirical therapy with new β-lactamase inhibitor combinations such as ceftazidime/avibactam, no class B carbapenemases were misclassified as class A carbapenemases. Both tests offer advantages. Used alone, without initial susceptibility tests, Rapidec Carba NP can provide positive results for some isolates after only 10 to 30 min incubation. BD Phoenix CPO Detect provides novel advantages such as automated carbapenemase detection, inclusion in susceptibility panels to eliminate delays and subjectivity in initiating carbapenemase tests, and classification of most carbapenemases.


[Hyperergic reactions to tuberculin].
Examination in 34875 children and adults revealed in 15.2% positive tuberculin reactions. In 0.23% these reactions were of hyperergic character. The authors present a detailed clinico-roentgenological characterization of these hyperergic reaction. The effect was established of BCG vaccination, contact of patients with tuberculosis, chronic infection foci on the skin sensitivity to tuberculin.


[A gas-chromatographic method for the determination of carbamazepine, phenobarbital, phenytoin and primidone in the same extract of serum (author's transl)].
A gas-chromatographic method for the determination of the antiepileptic drugs carbamazepine, phenobarbital, phenytoin and primidone in the same extract of serum is presented. Saturated ammonium sulfate solution is added to 1 ml serum, followed by extraction with chloroform. The organic phase is separated and evaporated. The residue is dissolved in 100 microliter ethylacetate/acetic acid (100 ml + 1 ml) for gas-chromatography. The gas-chromatographic determination is carried out under isothermal conditions without derivatisation, using SP 2250 DA as stationary phase for the determination of phenobarbital and phenytoin, and Dexsil 300 for the determination of carbamazepine and primidone. The coefficient of variation for the precision from day to day ranges from 4.3 to 7.5%, the recovery from 93.5 to 111%. The specificity was proven by comparison with the relative retention times of about 100 drugs. The method is compared with the corresponding EMIT-tests.


Magic Rings: Navigation in the Ring Chemical Space Guided by the Bioactive Rings.
The large majority of bioactive molecules contain a more or less complex ring system as a central structural element. This central core determines the basic molecule shape, keeps substituents in their proper positions, and often also contributes to the biological activity itself. In this study the ring systems extracted from one billion molecules are processed and differences between rings from bioactive molecules and common synthetic molecules are analyzed. The bioactive rings seem to be distributed throughout the large portion of chemical space, but not uniformly; one can see several more dense regions, where the bioactive rings often appear in small clusters, as well as empty areas. A web tool offering an interactive navigation in the ring chemical space and supporting identification of bioisosteric ring analogs available at https://bit.ly/magicrings is also described.


The three-dimensional structure of aquaporin-1.
The entry and exit of water from cells is a fundamental process of life. Recognition of the high water permeability of red blood cells led to the proposal that specialized water pores exist in the plasma membrane. Expression in Xenopus oocytes and functional studies of an erythrocyte integral membrane protein of relative molecular mass 28,000, identified it as the mercury-sensitive water channel, aquaporin-1 (AQP1). Many related proteins, all belonging to the major intrinsic protein (MIP) family, are found throughout nature. AQP1 is a homotetramer containing four independent aqueous channels. When reconstituted into lipid bilayers, the protein forms two-dimensional lattices with a unit cell containing two tetramers in opposite orientation. Here we present the three-dimensional structure of AQP1 determined at 6A resolution by cryo-electron microscopy. Each AQP1 monomer has six tilted, bilayer-spanning alpha-helices which form a right-handed bundle surrounding a central density. These results, together with functional studies, provide a model that identifies the aqueous pore in the AQP1 molecule and indicates the organization of the tetrameric complex in the membrane.


The vulval vestibular mucosa-morphological effects of oral contraceptives and menstrual cycle.
An erythematous and hypersensitive vestibular mucosa has been observed during the use of combined oral contraceptives (COC). Hormonal effects on the vestibular morphology have not been studied. Our aim was to evaluate the morphology of the vulval vestibular mucosa during the influence of COC and during the menstrual cycle. Forty-five healthy women (20 using COC and 25 not using COC) were included. A 6-mm punch biopsy was obtained from the right posterior vestibule on days 7-11 of the menstrual cycle. A corresponding biopsy was taken 2 weeks later in 16 women without COC. The epithelial morphology was estimated by measuring interdermal papilla distance, dermal papilla to surface, from basal layer to surface and width of dermal papillae. A histopathological assessment was made. The vulval vestibular mucosa of women using COC displayed a larger distance between the dermal papillae (P = 0.04) and a larger space from the dermal papillae to the epithelial surface (P = 0.03) compared with controls in the follicular phase. Women without COC displayed a larger interdermal papilla distance in the luteal phase compared with the follicular phase, P = 0.02. Histopathology showed more superficial blood vessels in the COC users (P < 0.01). The vulval vestibular mucosa of women with COC display an altered morphological pattern with shallow and sparse dermal papillae compared with the follicular phase. Similar findings are seen in women without COC during the luteal phase which indicate a gestagenic effect on the mucosa. Associations between the morphological pattern and changes in mucosal mechanical sensitivity require further studies.


Desynchronization of autonomic response and central autonomic network connectivity in posttraumatic stress disorder.
Although dysfunctional emotion regulatory capacities are increasingly recognized as contributing to posttraumatic stress disorder (PTSD), little work has sought to identify biological markers of this vulnerability. Heart rate variability (HRV) is a promising biomarker that, together with neuroimaging, may assist in gaining a deeper understanding of emotion dysregulation in PTSD. The objective of the present study was, therefore, to characterize autonomic response patterns, and their related neuronal patterns in individuals with PTSD at rest. PTSD patients (N = 57) and healthy controls (N = 41) underwent resting-state fMRI. Connectivity patterns of key regions within the central autonomic network (CAN)-including the ventromedial prefrontal cortex (vmPFC), amygdala, and periaqueductal gray (PAG)-were examined using a seed-based approach. Observed connectivity patterns were then correlated to resting HRV. In contrast to controls, individuals with PTSD exhibited lower HRV. In addition, whereas controls engaged a localized connectivity pattern of CAN-related brain regions, in PTSD, key CAN regions were associated with widespread connectivity patterns in regions related to emotional reactivity (vmPFC and amygdala to insular cortex and lentiform nucleus; PAG to insula) and motor readiness (vmPFC and amygdala to precentral gyrus; PAG to precentral gyrus and cerebellum). Critically, whereas CAN connectivity in controls was strongly related to higher HRV (insula, mPFC, superior frontal cortex, thalamus), HRV covariation was absent in PTSD subjects. This study provides the first evidence for a specific psychophysiological-neuronal profile in PTSD individuals characterized by lower resting HRV and a lack of HRV covariation with CAN-related brain connectivity. Hum Brain Mapp 38:27-40, 2017. © 2016 Wiley Periodicals, Inc.


[Myeloid metaplasia with myelofibrosis associated with acute coronary syndrome. Case report].
A 66-year-old man with myelofibrosis and myeloid metaplasia was admitted to hospital where he was diagnosed with acute coronary syndrome. Percutaneous transluminal coronary recanalization followed by drug-eluting stenting resulted in successful revascularization. Because besides thrombus formation the myeloproliferative syndrome has a potential for bleeding, it is very important to carefully follow-up any clotting abnormality.


Endogenous endophthalmitis associated with intravenous drug abuse.
To describe demographics, characteristics, and management of eyes with endogenous endophthalmitis (EE) in intravenous drug abusers. Retrospective chart review of patients with EE who presented to University Hospital, New Jersey Medical School between January 2005 and October 2012. Thirty-four patients presented with a clinical diagnosis of EE during this time period. Eighteen patients (53%) had a history of intravenous drug abuse with at least 15 of the 18 patients reporting intravenous heroin abuse. Average age was 43 years (24-61 years), 13 were men. Three patients had bilateral involvement. Mean time from the onset of symptoms to presentation was 10 days (range, 1-42 days). Presenting vision in the affected eye ranged from 20/100 to no light perception; the majority of patients had vision of counting fingers or worse. All patients were admitted to the hospital for intravenous antimicrobials and further workup. Final best-corrected visual acuity at the last follow-up ranged from 20/20 to no light perception, with most patients having final vision of 20/400 or worse. We reviewed 21 eyes of 18 patients with EE and concurrent intravenous drug abuse over an 8-year period; our study shows extremely poor visual prognosis associated with EE in this setting.


Some ethical and legal issues in assisted reproductive technology.
The potential and actual applications of reproductive technologies have been reviewed by many governmental committees, and laws have been enacted in several countries to accommodate, limit and regulate their use. Regulatory systems have nevertheless left some legal and ethical issues unresolved, and have caused other issues to arise. Issues that regulatory systems leave unresolved, or that systems have created, include disposal of embryos that remain after patients' treatments are concluded, and multiple implantation and pregnancy. This may result in risks to maternal, embryonic and neonatal life and health, and the contentious relief that may be achieved by selective reduction of multiple pregnancies. A further concern arises when clinics must (or choose to) publicize their success rates, and they compete for favorable statistics by questionable patient selection criteria and treatment priorities.


Reappraising personal values in cancer: Meaning-in-life adaptation, meaningfulness, and quality of life.
This study explores the reappraisal of personal values among people with cancer post-diagnosis and its connections to meaningfulness, encompassing personal meaning and sense of meaning, and various dimensions of quality of life. A total of 144 patients with diverse cancer types and a control group comprising 158 healthy adults with similar demographic characteristics completed the Valued Living Questionnaire-Perceived Change, the Personal Meaning Profile-Brief, the Portrait Values Questionnaire, and the Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being. Cancer patients displayed a substantial values reappraisal compared to healthy adults. They reported an increased significance of social areas and self-transcendental values, including close relationships (family, partner, and friends), spirituality, citizenship, and universalism, as well as an increased importance of self-care and self in general. Conversely, cancer patients assigned less importance to work and self-enhancement values such as stimulation, power, and achievement. Cluster analysis revealed that patients who reappraised their values exhibited higher meaningfulness and better indicators of quality of life, including spiritual and physical well-being, compared to patients who did not modify their value system. These findings underscore the importance of assessing and promoting meaning-in-life adaptability among people with cancer, with potential applications in meaning-centered therapies and interventions aimed at enhancing psychological flexibility.


Falls in stroke patients: risk factors and risk management.
Falls are common events among hospital inpatients and constitute a major health problem in the rehabilitation setting. Many risk factors for falls have been identified for stroke patients, such as muscle weakness, medication side effects, hypoglycemia, hypotension, etc. To assess the risk factors for falls among patients hospitalized for rehabilitation following acute stroke. In a retrospective study of 56 falls over a period of 5 years in 41 stroke patients hospitalized for rehabilitation we surveyed the nurses' safety risk assessment of the fall. Thirty patients fell once, 9 patients twice and 2 patients four times. The data were obtained from the medical and nursing records. Safety precautions were taken by the nurses for the entire group of patients. Most of the falls occurred among male patients who had reduced muscular tone (70%), paralysis (54%) and/ or hypoesthesia in the involved side of the body. Patients who suffered from hemiplegia fell more often than those with hemiparesis (Wilcoxon rank sum test, P = 0.04, one-sided). Forty-eight percent of the falls occurred during the first month after the last stroke, 70% during the morning or the afternoon, and 62% occurred close to the patient's bed. In 89% of falls the patients used hypoglycemic, antihypertensive, tranquilizing or neuroleptic drugs. Communication disorders (29%), hemianopia or blindness (21%) and visuospatial agnosia (18%) were incremental risk factors for falls. Fifty percent of the falls were caused by either an intrinsic or extrinsic mechanism. These data suggest that the group of stroke patients at risk for falls in a rehabilitation department can be identified by a variety of impairment and functional assessments. The information may be potentially useful for designing interventions directed at reducing fall frequency among stroke survivors.


[Antifungal and antimicrobial activity of beta-ionone and vitamin A derivatives].
The antifungal and antimicrobic activity of some derivatives of beta-ionon and vitamin A was studied. These compounds (citral, pseudo-ionon. beta-ionon aldehyde C14, ketone C18 and its derivatives--4,18-diketone, alcohol C18, semicarbazide ketone C18), as well as vitamin A and its derivatives--retinal, acetate, retinoic acid--differ in composition, structure and substituents of C-atoms in beta-ionon ring and in polyenoic chain. Fusarium solani, Botrytis cenerea and Verticillum dahliae II, race 447 were used as test organisms when studying the antifungal activity. When studying the antimicrobic activity, ketone C18 and alcohol C18 were tested using the museum strains Staphylococcus aureus 209 P, hemolytic Streptococcus pyogenes FF-38, Streptococcus sp. TOM-1606, Micrococcus luteus 2665, as well as the strains Staphylococcus aureus isolated from pyoderma patients. The antifungal activity was determined by inhibition of spore germination, and the antimicrobic activity--by the value of areas of growth inhibition of the test organisms on the agar medium. All the compounds mentioned above possessed the antifungal activity against all the phytopathogenes used. The degree of this activity depends on the composition and structure of both the cyclic part and polyenoic chain. on the number of conjugated unsaturations, substituents and end groups of C-atom. Ketone C18 is the most active one. It inhibits spore germination by 100-94% at concentrations ranging from 0.05 to 0.005% and 24 h exposure, and by 100% at the concentration of 0.1% and 72 h exposure. Alcohol C18 possesses almost the same antifungul activity.(ABSTRACT TRUNCATED AT 250 WORDS)


Differential activation of rod outer segment membrane guanylate cyclases, ROS-GC1 and ROS-GC2, by CD-GCAP and identification of the signaling domain.
The ROS-GC is one of the two subfamilies of membrane guanylate cyclases. It distinguishes itself from the other surface receptor subfamily in that its members are not regulated by extracellular peptides; instead, they are modulated by intracellular Ca2+ signals. There are two members of the subfamily, ROS-GC1 and ROS-GC2. An intriguing feature of ROS-GC1 is that it has two Ca2+ switches. One switch inhibits the enzyme at micromolar concentrations of Ca2+, and the other stimulates. The inhibitory switch is linked to phototransduction, and it is likely that the stimulatory switch is linked to retinal synaptic activity. Ca2+ acts indirectly via Ca(2+)-binding proteins, GCAPs and CD-GCAP. GCAPs modulate the inhibitory switching component of the cyclase and CD-GCAP turns on the activation signaling switch. The activating switch of ROS-GC2 has not so far been scrutinized. The present study shows that CD-GCAP is linked to the activation signaling switch of ROS-GC2, but the linkage is about 10-fold weaker than that of the ROS-GC1. Thus, CD-GCAP is a specific ROS-GC1 activator. Furthermore, through a series of expression studies on the mutants involving deletion, building of hybrids, and reconstruction of a heterologous cyclase, the study confirms that the CD-GCAP regulated switch resides within the amino acid segment 736-1053 of the cyclase.


Effect of verapamil on home self-measurement of blood pressure and heart rate by hypertensive patients. Verapamil-Frequency Research Group.
Achievement of a controlled blood pressure and improvement of cardiovascular risk profile are the mainstays of therapy for hypertension. To assess the responses of heart rate and blood pressure in hypertensive patients to 240 mg/day sustained release verapamil. We assessed the effect of 240 mg/day sustained released verapamil on blood pressure and heart rate, measured in the office and at home, in 1395 hypertensive outpatients with mild-to-moderate hypertension, who were using an Omron HEM 705 CP automatic device for self-measurement. The period of observation was 3 months. Blood pressure decreased both in the medical office and at the patient's home, the measurements obtained at home being lower than those found in the office. Heart rate decreased in a significant and particular way. The decrease was greatest among those patients with histories of myocardial infarction and among relatively young patients, who exhibit a tendency towards higher than normal baseline heart rates. Overall, there was a shift of the heart-rate curve towards more controlled levels clustered around heart rates between 65 and 75 beats/min. Home self-measurement showed that the data gathered by the patients at home are reliable and that, when cut-off values of 140/90 mm Hg for blood pressure are used, the percentage of patients with controlled blood pressures is 62%, whereas the percentage obtained in the medical office by the physician is 56%. If cut-off values of 135/85 mm Hg are considered for self-measurements at home, according to the VI JNC recommendations, the percentage of patients with controlled blood pressures is 25.4%.


Molecular features of product release for the PKA catalytic cycle.
Although ADP release is the rate limiting step in product turnover by protein kinase A, the steps and motions involved in this process are not well resolved. Here we report the apo and ADP bound structures of the myristylated catalytic subunit of PKA at 2.9 and 3.5 Å resolution, respectively. The ADP bound structure adopts a conformation that does not conform to the previously characterized open, closed, or intermediate states. In the ADP bound structure, the C-terminal tail and Gly-rich loop are more closed than in the open state adopted in the apo structure but are also much more open than the intermediate or closed conformations. Furthermore, ADP binds at the active site with only one magnesium ion, termed Mg2 from previous structures. These structures thus support a model where ADP release proceeds through release of the substrate and Mg1 followed by lifting of the Gly-rich loop and disengagement of the C-terminal tail. Coupling of these two structural elements with the release of the first metal ion fills in a key step in the catalytic cycle that has been missing and supports an ensemble of correlated conformational states that mediate the full catalytic cycle for a protein kinase.


Conformal Lithium Fluoride Protection Layer on Three-Dimensional Lithium by Nonhazardous Gaseous Reagent Freon.
Research on lithium (Li) metal chemistry has been rapidly gaining momentum nowadays not only because of the appealing high theoretical capacity, but also its indispensable role in the next-generation Li-S and Li-air batteries. However, two root problems of Li metal, namely high reactivity and infinite relative volume change during cycling, bring about numerous other challenges that impede its practical applications. In the past, extensive studies have targeted these two root causes by either improving interfacial stability or constructing a stable host. However, efficient surface passivation on three-dimensional (3D) Li is still absent. Here, we develop a conformal LiF coating technique on Li surface with commercial Freon R134a as the reagent. In contrast to solid/liquid reagents, gaseous Freon exhibits not only nontoxicity and well-controlled reactivity, but also much better permeability that enables a uniform LiF coating even on 3D Li. By applying a LiF coating onto 3D layered Li-reduced graphene oxide (Li-rGO) electrodes, highly reduced side reactions and enhanced cycling stability without overpotential augment for over 200 cycles were proven in symmetric cells. Furthermore, Li-S cells with LiF protected Li-rGO exhibit significantly improved cyclability and Coulombic efficiency, while excellent rate capability (∼800 mAh g-1 at 2 C) can still be retained.


The Americans With Disabilities Act and dermatologists.
The passage and implementation of the Americans With Disabilities Act of 1990 (ADA) has established certain rights in law for persons with disabilities, including patients with skin disease. For workers in workplaces employing more than 15 workers, management may not exclude a worker from a job unless it can be objectively demonstrated that their skin disease will make it impossible for them to perform the essential functions of the job. Employers, at their expense, are now required to provide those disabled with skin disease with reasonable accommodation to allow them to perform their jobs while having their skin impairment. Unless the skin impairment can be shown to place the worker or their fellow workers at material risk to their health, the employer may not exclude them from working--even if the employer is concerned that it may make their skin disease worse. The act applies both to new employees and those acquiring disabling skin disease who are employed and wish to stay at work or return to work. Arbitration of dispute between employees and management are the responsibility of the Equal Employment Opportunities Commission (EEOC), a federal agency responsible for the legislation.


Comparison of human optimized bacterial luciferase, firefly luciferase, and green fluorescent protein for continuous imaging of cell culture and animal models.
Bioluminescent and fluorescent reporter systems have enabled the rapid and continued growth of the optical imaging field over the last two decades. Of particular interest has been noninvasive signal detection from mammalian tissues under both cell culture and whole animal settings. Here we report on the advantages and limitations of imaging using a recently introduced bacterial luciferase (lux) reporter system engineered for increased bioluminescent expression in the mammalian cellular environment. Comparison with the bioluminescent firefly luciferase (Luc) system and green fluorescent protein system under cell culture conditions demonstrated a reduced average radiance, but maintained a more constant level of bioluminescent output without the need for substrate addition or exogenous excitation to elicit the production of signal. Comparison with the Luc system following subcutaneous and intraperitoneal injection into nude mice hosts demonstrated the ability to obtain similar detection patterns with in vitro experiments at cell population sizes above 2.5 × 10(4) cells but at the cost of increasing overall image integration time.


Evolutionary diversification of pigment pattern in Danio fishes: differential fms dependence and stripe loss in D. albolineatus.
The developmental bases for species differences in adult phenotypes remain largely unknown. An emerging system for studying such variation is the adult pigment pattern expressed by Danio fishes. These patterns result from several classes of pigment cells including black melanophores and yellow xanthophores, which differentiate during metamorphosis from latent stem cells of presumptive neural crest origin. In the zebrafish D. rerio, alternating light and dark horizontal stripes develop, in part, owing to interactions between melanophores and cells of the xanthophore lineage that depend on the fms receptor tyrosine kinase; zebrafish fms mutants lack xanthophores and have disrupted melanophore stripes. By contrast, the closely related species D. albolineatus exhibits a uniform pattern of melanophores, and previous interspecific complementation tests identified fms as a potential contributor to this difference between species. Here, we survey additional species and demonstrate marked variation in the fms-dependence of hybrid pigment patterns, suggesting interspecific variation in the fms pathway or fms requirements during pigment pattern formation. We next examine the cellular bases for the evolutionary loss of stripes in D. albolineatus and test the simplest model to explain this transformation, a loss of fms activity in D. albolineatus relative to D. rerio. Within D. albolineatus, we demonstrate increased rates of melanophore death and decreased melanophore migration, different from wild-type D. rerio but similar to fms mutant D. rerio. Yet, we also find persistent fms expression in D. albolineatus and enhanced xanthophore development compared with wild-type D. rerio, and in stark contrast to fms mutant D. rerio. These findings exclude the simplest model in which stripe loss in D. albolineatus results from a loss of fms-dependent xanthophores and their interactions with melanophores. Rather, our results suggest an alternative model in which evolutionary changes in pigment cell interactions themselves have contributed to stripe loss, and we test this model by manipulating melanophore numbers in interspecific hybrids. Together, these data suggest evolutionary changes in the fms pathway or fms requirements, and identify changes in cellular interactions as a likely mechanism of evolutionary change in Danio pigment patterns.


Chemically synthesized peptides elicit neutralizing antibody to bovine enterovirus.
Synthetic peptides representing 14 regions of the bovine enterovirus structural proteins were used to raise antibodies in mice. The peptides were predicted using amino acid sequence alignments with the position of antigenic sites on other picornaviruses. Five of the anti-peptide antibodies reacted with the virus in an immunoprecipitation test. Furthermore, each of these anti-peptide antibodies neutralized virus infectivity; those directed against peptides of VP2 and VP3 neutralized to a greater extent than those directed against peptides of VP1. The positions of these epitopes in the viral structural proteins are discussed in relation to corresponding positions in other picornaviruses.


Biological markers of male reproductive toxicology.
Reproduction is a complex, stepwise series of processes that begins with gametogenesis, continues through gamete interaction, implantation, embryonic development, growth, parturition, and postnatal adaptation, and is completed with the development and sexual maturation of the newly formed organism. These reproductive processes do not take place in a chemically pristine environment, but rather in an environment increasingly contaminated with the products and by-products of the chemical age in which we live. Some environmental pollutants are known to be carcinogenic, mutagenic, or toxic to the reproductive system, but most have not been adequately tested for reproductive toxicity. Just as reproduction is complex, biological mechanisms underlying toxicology are similarly complex and involve absorption, distribution, metabolism (toxification and/or detoxification), excretion, and repair. The synthesis of these sciences into the relatively nascent science of reproductive toxicology includes teratology, pharmacology, epidemiology, and occupational and environmental health. Female reproductive function (especially pregnancy outcome) has historically been the focus of attention, but there is increasing interest in the effects of chemical exposure on male reproductive function. Several reports have documented the physiology, biochemistry, and toxicology of male mammalian reproduction, and evaluated susceptibility of the male to the effects of exogenous chemicals.


In defence of mandatory bicycle helmet legislation: response to Hooper and Spicer.
We invoke a triple rationale to rebut Hooper and Spicer's argument against mandatory helmet laws. First, we use the laws of physics and empirical studies to show how bicycle helmets afford substantial protection to the user. We show that Hooper and Spicer erroneously downplay helmet utility and that, as a result, their attack on the utilitarian argument for mandatory helmet laws is weakened. Next, we refute their claim that helmet legislation comprises unjustified paternalism. We show the healthcare costs of bareheaded riding to pose significant third party harms. It follows, we argue, that a utilitarian case for helmet laws can be sustained by appeal to Mill's Harm Principle. Finally, we reject Hooper and Spicer's claim that helmet laws unjustly penalise cyclists for their own health-affecting behaviour. Rather, we show their argument to suffer by disanalogy with medical cases where injustice may be more evident, for example, denial of bypass surgery to smokers. We conclude that mandatory helmet laws offer substantial utility and are entirely defensible within the framework of a liberal democracy.


Phytochelatins, a class of heavy-metal-binding peptides from plants, are functionally analogous to metallothioneins.
Phytochelatins are a class of heavy-metal-binding peptides previously isolated from cell suspension cultures of several dicotyledonous and monocotyledonous plants. These peptides consist of repetitive gamma-glutamylcysteine units with a carboxyl-terminal glycine and range from 5 to 17 amino acids in length. In the present paper we show that all plants tested synthesized phytochelatins upon exposure to heavy metal ions. No evidence for the occurrence of metallothionein-like proteins was found. All data so far obtained indicate that phytochelatins are involved in detoxification and homeostasis of heavy metals in plants and thus serve functions analogous to those of metallothioneins in animals and some fungi. Phytochelatins are induced by a wide range of metal anions and cations. Phytochelatin synthesis in suspension cultures was inhibited by buthionine sulfoximine, a specific inhibitor of gamma-glutamylcysteine synthetase (EC 220.127.116.11). This finding and kinetic studies of phytochelatin induction point to a synthesis from glutathione or its precursor, gamma-glutamylcysteine, in a sequential manner, thereby generating the set of homologous peptides.


Gambling Participation and Problem Gambling Severity in a Stratified Random Survey: Findings from the Second Social and Economic Impact Study of Gambling in Tasmania.
Demographic characteristics associated with gambling participation and problem gambling severity were investigated in a stratified random survey in Tasmania, Australia. Computer-assisted telephone interviews were conducted in March 2011 resulting in a representative sample of 4,303 Tasmanian residents aged 18 years or older. Overall, 64.8% of Tasmanian adults reported participating in some form of gambling in the previous 12 months. The most common forms of gambling were lotteries (46.5%), keno (24.3%), instant scratch tickets (24.3%), and electronic gaming machines (20.5%). Gambling severity rates were estimated at non-gambling (34.8%), non-problem gambling (57.4%), low risk gambling (5.3%), moderate risk (1.8%), and problem gambling (.7%). Compared to Tasmanian gamblers as a whole significantly higher annual participation rates were reported by couples with no children, those in full time paid employment, and people who did not complete secondary school. Compared to Tasmanian gamblers as a whole significantly higher gambling frequencies were reported by males, people aged 65 or older, and people who were on pensions or were unable to work. Compared to Tasmanian gamblers as a whole significantly higher gambling expenditure was reported by males. The highest average expenditure was for horse and greyhound racing ($AUD 1,556), double the next highest gambling activity electronic gaming machines ($AUD 767). Compared to Tasmanian gamblers as a whole problem gamblers were significantly younger, in paid employment, reported lower incomes, and were born in Australia. Although gambling participation rates appear to be falling, problem gambling severity rates remain stable. These changes appear to reflect a maturing gambling market and the need for population specific harm minimisation strategies.


Two classes of ouabain binding sites in ferret heart and two forms of Na+-K+-ATPase.
In partially purified Na+-K+-adenosinetriphosphatase (ATPase) obtained from ferret heart, ouabain produced a monophasic inhibition curve; however, the curve spanned over 5 logarithmic units, indicating the presence of more than one classes of enzyme. [3H]ouabain binding studies revealed high-and low-affinity binding sites in approximately equal abundance, with apparent dissociation constants of 10 and 230 nM, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of phosphoenzyme formed from [gamma-32P]ATP showed two distinct K+-sensitive bands of approximately 100,000 molecular weight. Phosphoenzyme formation from the high-molecular-weight alpha(+) form was selectively inhibited by N-ethylmaleimide. Ouabain caused a 50% inhibition of phosphorylation of the alpha(+) form at 40 nM and the lower-molecular-weight alpha form at 300 nM. In papillary muscle preparations, 1-30 nM ouabain produced a modest positive inotropic effect that reached an apparent plateau at 30 nM. Further increases in ouabain concentrations, however, produced additional and prominent inotropic effects at 0.1-10 microM. These results indicate for the first time in cardiac muscle that the high- and low-affinity ouabain binding sites are associated with the alpha(+) and alpha forms of the Na+-K+-ATPase, respectively, and that binding of ouabain to either of these sites causes enzyme inhibition and the positive inotropic effect.


Adult age differences in frontostriatal representation of prediction error but not reward outcome.
Emerging evidence from decision neuroscience suggests that although younger and older adults show similar frontostriatal representations of reward magnitude, older adults often show deficits in feedback-driven reinforcement learning. In the present study, healthy adults completed reward-based tasks that did or did not depend on probabilistic learning, while undergoing functional neuroimaging. We observed reductions in the frontostriatal representation of prediction errors during probabilistic learning in older adults. In contrast, we found evidence for stability across adulthood in the representation of reward outcome in a task that did not require learning. Together, the results identify changes across adulthood in the dynamic coding of relational representations of feedback, in spite of preserved reward sensitivity in old age. Overall, the results suggest that the neural representation of prediction error, but not reward outcome, is reduced in old age. These findings reveal a potential dissociation between cognition and motivation with age and identify a potential mechanism for explaining changes in learning-dependent decision making in old adulthood.


Differential characteristics of the electrocardiogram in early repolarization and acute pericarditis.
Electrocardiographic mimicry necessitates differentiation between acute pericarditis and early repolarization. Among 96 persons, limb-lead RST deviations occurred in all 48 with pericarditis, but only 27 with early repolarization. RS-T vectors (A RS-T) in pericarditis tended to be horizontal (25 patients) and left of the T vector, A T (28 patients); in early repolarization, A RS-T was vertical in 15 subjects and right of A T (20 subjects); P less than or equal to 0.01. RS-T depression in Lead V1 was more common in pericarditis (14 vs. two); isoelectric RS-T in Lead V6 was more common in early repolarization (10 vs. one); P less than or equal to 0.01. PR segment deviations occurred in both limb and precordial leads in pericarditis; in early repolarization they were confined to either lead group. Thus, R-ST deviations in both limb and precordial leads with horizontal A RS-T to left of A T and RS-T depression in Lead V1 favor pericarditis; vertical A RS-T and isoelectric RS-T in Lead V6 favor early repolarization.


Progress and Prospects in Gender Visibility at SMBE Annual Meetings.
Reduced visibility of women in science is thought to be one of the causes of their underrepresentation among scientists, in particular at senior positions. Visibility is achieved through publications, and through conference attendance and presentations. Here, we investigated gender differences in visibility at the annual meetings of the Society of Molecular Biology and Evolution. The analysis of meeting programs showed a regular increase in female speakers for the last 16 years. Data on abstract submission suggest that there are no gender-related preferences in the acceptance of contributed presentations at the most recent meetings. However, data collected on-site in 2015 and 2016 show that women asked only ∼25% of the questions, that is, much less than expected given the female attendance. Understanding the reasons for this pattern is necessary for the development of policies that aim to reduce imbalance in visibility.


Higher Thioguanine Nucleotide Metabolite Levels are Associated With Better Long-term Outcomes in Patients With Inflammatory Bowel Diseases.
The aim of this study was to assess whether sustained 6-thioguaninenucleotide (6-TGN) levels were associated with improved long-term outcomes in patients with inflammatory bowel diseases (IBD). Cross-sectional data have shown that thiopurine metabolites are correlated with clinical efficacy in patient receiving thiopurines for IBD but the role for serial measurements through treatment course is unclear. We conducted a retrospective cohort study including patients with IBD on thiopurine monotherapy and had serial 6-TGN levels measured. Predictive variables included demographics, disease phenotype, 6-TGN levels (nadir, median, and peak levels). The primary outcome was the development of a disease relapse. The secondary outcome was the need for IBD surgery. Two hundred eighteen 6-TGN samples from 87 patients were analyzed. Nadir and median 6-TGN levels were significantly higher in patients who did not relapse [185 and 233 pmol per 8×10 red blood cells (RBCs)] as compared with levels in patients who did relapse (150 and 167 pmol per 8×10 RBCs, P=0.025) but there was no significant difference in peak 6-TGN level. When adjusted for confounding factors, a nadir 6-TGN level ≥161 and a median 6-TGN level ≥264 were associated with a significant decrease in the rate of disease exacerbation (hazard ratio: 0.5; 95% confidence interval, 0.26-0.87; P=0.016 and hazard ratio: 0.4; 95% confidence interval, 0.2-0.82; P=0.14). Serial thiopurine metabolite level assessments and dose adjustment aiming to maintain higher 6-TGN levels may be helpful to improve long-term outcomes in patients with IBD.


Os parietale partitum: Exploring the prevalence of this trait in four contemporary populations.
Os parietale partitum is a variable segmentation of the parietal bone. This manifests as a parietal division in the anteroposterior or superoinferior planes that is separated by an unusual suture and can be complete or incomplete. The existence of parietal divisions was observed and documented more than 260 years ago. The main objectives of this paper are to record the incidence of this rare trait in four modern populations with no previous records of it and provide a review of the literature. Four contemporary skeletal collections from Crete (Greece), Limassol (Cyprus), Coimbra (Portugal) and Salvador (Brazil) were assessed by the authors of this paper for non-metric cranial traits. Out of 711 skulls, only three cases of parietal division were found and all three originated from the Cypriot collection. These three cases were anatomically analyzed, showing that all three cases were adult females and showed unilateral expression of the trait. Two skulls showed superoinferior division, and the third case showed anteroposterior division. Numerous other cranial non-metric traits were found in these three skulls. Based on the cemetery archives, there seems to be no genetic link between the individuals bearing this trait. Further genetic analysis is suggested in order to verify this conclusion.