Reentrant ventricular arrhythmias in the late myocardial infarction period in the dog. 13. Correlation of activation and refractory maps.
Isochronal maps of ventricular activation were analyzed in dogs 3-5 days after ligation of the left anterior descending coronary artery utilizing a 64-channel multiplexer. Isochronal maps of the effective refractory period were determined from 62 epicardial sites and correlated with the activation maps. The ischemia occurring in the surviving epicardial layer prolonged refractoriness in a spatially nonuniform manner. The resulting pattern of refractoriness on the epicardial surface resembled concentric rings of isorefractoriness which increased in duration from the normal zone to the center of the ischemic zone. The formation of an arc of functional unidirectional conduction block occurred along the gradient of refractoriness and the exact location of the arc depended on the S1-S2 interval. When a short S1-S2 failed to induce reentry, fewer adjacent sites with sufficiently disparate refractoriness formed a smaller arc of block. A subsequent S3 encountered further nonuniformly shortened refractoriness (normal areas had shortened refractoriness greater than ischemic areas) and the arc of block was lengthened. This required a longer time for the wavefront to circulate around the arc. When it then reached the distal side of the arc, refractoriness had expired proximal to the arc and reentry occurred. Similarly, nonuniform shortening of refractoriness explained why one reentrant beat may or may not produce successive reentrant beats. Therefore, the spatial pattern of refractoriness forms the substrate for the arc of unidirectional conduction block that is fundamental to the development of ventricular reentry in this experimental model.


The influence of the cryopreserving process on rubidium uptake by human spermatozoa.
The effect of cryopreservation and its phases on the netto-Rb+ uptake (analogous to the K+ uptake) of washed human spermatozoa was investigated under different conditions. The Rb+ uptake reached a steady state after almost 50-min incubation both before and after the cryopreserving process. The Rb+ uptake could be reduced to about 30% by ouabain and to about 80% by monoiodoacetate. The cryoprotective agents glycerol (G), ethylene glycol (EG), and particularly dimethylsulfoxide (DMSO) reduced the Rb+ uptake by a factor dependent on its concentration. The comparative cryopreservation with G, EG, DMSO,and dimethylformamide (DMFA) as cryoprotective agents caused an inhibition especially of the ouabain-sensitive netto-Rb+ uptake. This inhibitory effect became stronger from G to EG and from DMSO to DMFA. Within the course of cryopreservation especially the process of pelletation, Rb+ uptake was reduced, whereas the prolongation of the equilibration time up to 180 min moderated the effect of the cryopreservation on the Rb+ uptake. Beside the reduction of the absolute Rb+ uptake, the cryopreservation altered the dependence of the netto-Rb+ uptake on the extracellular Rb+ concentration in a sigmoid manner.


Strategies for knowledge acquisition.
This paper describes knowledge acquisition strategies developed in the course of handcrafting a diagnostic system and reports on their consequent implementation in MORE, an automated knowledge acquisition system. We describe MORE in some detail, focusing on its representation of domain knowledge, rule generation capabilities, and interviewing techniques. MORE's approach is shown to embody methods which may prove fruitful to the development of knowledge acquisition systems in other domains.


Rhamnogalacturonan II structure shows variation in the side chains monosaccharide composition and methylation status within and across different plant species.
A paradigm regarding rhamnogalacturonans II (RGII) is their strictly conserved structure within a given plant. We developed and employed a fast structural characterization method based on chromatography and mass spectrometry, allowing analysis of RGII side chains from microgram amounts of cell wall. We found that RGII structures are much more diverse than so far described. In chain A of wild-type plants, up to 45% of the l-fucose is substituted by l-galactose, a state that is seemingly uncorrelated with RGII dimerization capacity. This led us to completely reinvestigate RGII structures of the Arabidopsis thaliana fucose-deficient mutant mur1, which provided insights into RGII chain A biosynthesis, and suggested that chain A truncation, rather than l-fucose to l-galactose substitution, is responsible for the mur1 dwarf phenotype. Mass spectrometry data for chain A coupled with NMR analysis revealed a high degree of methyl esterification of its glucuronic acid, providing a plausible explanation for the puzzling RGII antibody recognition. The β-galacturonic acid of chain A exhibits up to two methyl etherifications in an organ-specific manner. Combined with variation in the length of side chain B, this gives rise to a family of RGII structures instead of the unique structure described up to now. These findings pave the way for studies on the physiological roles of modulation of RGII composition.


The evolution of substrate specificity-associated residues and Ca(2+) -binding motifs in EF-hand-containing type II NAD(P)H dehydrogenases.
Most eukaryotic organisms, except some animal clades, have mitochondrial alternative electron transport enzymes that allow respiration to bypass the energy coupling in oxidative phosphorylation. The energy bypass enzymes in plants include the external type II NAD(P)H dehydrogenases (DHs) of the NDB family, which are characterized by an EF-hand domain for Ca(2+) binding. Here we investigate these plant enzymes by combining molecular modeling with evolutionary analysis. Molecular modeling of the Arabidopsis thaliana AtNDB1 with the yeast ScNDI1 as template revealed distinct similarities in the core catalytic parts, and highlighted the interaction between the pyridine nucleotide and residues correlating with NAD(P)H substrate specificity. The EF-hand domain of AtNDB1 has no counterpart in ScNDI1, and was instead modeled with Ca(2+) -binding signal transducer proteins. Combined models displayed a proximity of the AtNDB1 EF-hand domain to the substrate entrance side of the catalytic part. Evolutionary analysis of the eukaryotic NDB-type proteins revealed ancient and recent reversions between the motif observed in proteins specific for NADH (acidic type) and NADPH (non-acidic type), and that the clade of enzymes with acidic motifs in angiosperms derives from non-acidic-motif NDB-type proteins present in basal plants, fungi and protists. The results suggest that Ca(2+) -dependent external NADPH oxidation is an ancient process, indicating that it has a fundamental importance for eukaryotic cellular redox metabolism. In contrast, the external NADH DHs in plants are products of a recent expansion, mirroring the expansion of the alternative oxidase family.


[Methodological approach to genetic-selection analysis of social behavior of animals (Using as an example domestic behavior of the silver fox)].
Social behaviour of animals, i.e. interspecies contacts, were studied in a population of silver foxes in the longitudinal model domestication experiment. Three social behaviour traits were analysed: contactibility, critical distance and domestication index. The latter trait is a linear function of elementary behaviour reactions, postures induced by man which is determined by the principal components method. The most valiable selection phenotype for further modelling of domestication, as shown by the capacity to adequately reflect a pattern of genotype--environment interaction, the structure of correlation ratios for different forms of behaviour and the level of genetic relationships between relatives, is the domestication index.


Antithrombotic treatment after stroke due to intracerebral haemorrhage.
Survivors of stroke due to intracerebral haemorrhage (ICH) are at risk of thromboembolism. Antithrombotic (antiplatelet or anticoagulant) treatments may lower the risk of thromboembolism after ICH, but they may increase the risks of bleeding. To determine the overall effectiveness and safety of antithrombotic drugs for people with ICH. We searched the Cochrane Stroke Group Trials Register (24 March 2017). We also searched the Cochrane Central Register of Controlled Trials (CENTRAL: the Cochrane Library 2017, Issue 3), MEDLINE Ovid (from 1948 to March 2017), Embase Ovid (from 1980 to March 2017), and online registries of clinical trials (8 March 2017). We also screened the reference lists of included trials for additional, potentially relevant studies. We selected all randomised controlled trials (RCTs) of any antithrombotic treatment after ICH. Three review authors independently extracted data. We converted categorical estimates of effect to the risk ratio (RR) or odds ratio (OR), as appropriate. We divided our analyses into short- and long-term treatment, and used fixed-effect modelling for meta-analyses. Three review authors independently assessed the included RCTs for risks of bias and we created a 'Summary of findings' table using GRADE. We included two RCTs with a total of 121 participants. Both RCTs were of short-term parenteral anticoagulation early after ICH: one tested heparin and the other enoxaparin. The risk of bias in the included RCTs was generally unclear or low, with the exception of blinding of participants and personnel, which was not done. The included RCTs did not report our chosen primary outcome (a composite outcome of all serious vascular events including ischaemic stroke, myocardial infarction, other major ischaemic event, ICH, major extracerebral haemorrhage, and vascular death). Parenteral anticoagulation did not cause a statistically significant difference in case fatality (RR 1.25, 95% confidence interval (CI) 0.38 to 4.07 in one RCT involving 46 participants, low-quality evidence), ICH, or major extracerebral haemorrhage (no detected events in one RCT involving 75 participants, low-quality evidence), growth of ICH (RR 1.64, 95% CI 0.51 to 5.29 in two RCTs involving 121 participants, low-quality evidence), deep vein thrombosis (RR 0.99, 95% CI 0.49 to 1.96 in two RCTs involving 121 participants, low quality evidence), or major ischaemic events (RR 0.54, 95% CI 0.23 to 1.28 in two RCTs involving 121 participants, low quality evidence). There is insufficient evidence from RCTs to support or discourage the use of antithrombotic treatment after ICH. RCTs comparing starting versus avoiding antiplatelet or anticoagulant drugs after ICH appear justified and are needed in clinical practice.


Central Nervous System Lymphoma.
This article reviews the clinical presentation, diagnostic workup, staging, and treatment of primary central nervous system (CNS) lymphoma and common manifestations of secondary CNS lymphoma. Lymphoma can arise in the CNS de novo (primary CNS lymphoma) or as the result of systemic disease (secondary CNS lymphoma). Symptoms may include focal neurologic deficits related to the disease site, cognitive decline, and symptoms of increased intracranial pressure. Standard treatment may differ based on lymphoma subtype and location. A majority of CNS lymphoma is diffuse large B-cell subtype and exhibits aggressive behavior. First-line treatment is generally methotrexate-based polychemotherapy. Response rates to treatment are high, approximately 80% to 90% for primary CNS lymphoma, but relapse is common. Consolidation approaches including myeloablative chemotherapy followed by autologous stem cell rescue, nonmyeloablative chemotherapy, radiation, and medical maintenance regimens reduce rates of relapse. The recent development of targeted agents such as Bruton tyrosine kinase inhibitors and immunomodulatory strategies have shown promise in the treatment of CNS lymphoma. Immunotherapy in the form of checkpoint inhibitors and chimeric antigen receptor T cells is being studied. More indolent forms of lymphoma may be treated with radiation or targeted therapy. CNS lymphoma is an uncommon but clinically meaningful manifestation of extranodal lymphoma. The diagnosis requires a high level of suspicion for rapid initiation of potentially curative treatment.


Primary testicular lymphoma demonstrates overexpression of the Wilms tumor 1 gene and different mRNA and miRNA expression profiles compared to nodal diffuse large B-cell lymphoma.
Diffuse large B-cell lymphoma (DLBCL) shows a high degree of clinical and biological heterogeneity. Primary testicular lymphoma (PTL) is an extranodal variant of DLBCL associated with a higher risk of recurrence, including contralateral testicles and central nervous system sanctuary sites. Several molecular aberrations, including somatic mutation of MYD88, CD79B, and upregulation of NF-kB, PDL-1, and PDL-2, are thought to contribute to the pathogenesis and poor prognosis of PTL. However, additional biomarkers are needed that may improve the prognosis and help understand the PTL biology and lead to new therapeutic targets. RNA from diagnostic tissue biopsies of the PTL-ABC subtype and matched nodal DLBCL-ABC subtype patients was evaluated by mRNA and miRNA expression. Screening of 730 essential oncogenic genes was performed, and their epigenetic connections were examined using the nCounter PAN-cancer pathway, and Human miRNA assays with the nCounter System (NanoString Technologies). PTL and nodal DLBCL patients were comparable in age, gender, and putative cell of origin (p > 0.05). Wilms tumor 1 (WT1) expression in PTL exceeded that in nodal DLBCL (>6-fold; p = 0.01, FDR <0.01) and WT1 associated pathway genes THBS4, PTPN5, PLA2G2A, and IFNA17 were upregulated in PTL (>2.0-fold, p < 0.01, FDR <0.01). Additionally, miRNAs targeting WT1 (hsa15a-5p, hsa-miR-16-5p, has-miR-361-5p, has-miR-27b-3p, has-miR-199a-5p, has-miR-199b-5p, has-miR-132-3p, and hsa-miR-128-3p) showed higher expression in PTL compared to nodal DLBCL (≥2.0-fold; FDR 0.01). Lower expression of BMP7, LAMB3, GAS1, MMP7, and LAMC2 (>2.0-fold, p < 0.01) was observed in PTL compared to nodal DLBCL. This research revealed higher WT1 expression in PTL relative to nodal DLBCL, suggesting that a specific miRNA subset may target WT1 expression and impact the PI3k/Akt pathway in PTL. Further investigation is needed to explore WT1's biological role in PTL and its potential as a therapeutic target.


Medication-overuse headache: The effect of a patient educational programme-A randomized controlled trial.
Little is known about the effects of non-pharmacological interventions among medication-overuse headache (MOH) patients, although non-pharmacological approaches combined with pharmacological treatment are recommended. The objective was to evaluate the effect of an educational programme as an add-on to standard treatment. Medication-overuse headache patients were randomized (1:1) in a single-centre setting to standard treatment with 12 weeks of education (I-group) versus standard treatment (C-group). The primary outcome was measurement of reduction in headache days/last month at 9 months' follow-up. Secondary outcomes were headache intensity, acute medication intake, bothersomeness, disability, physical activity and patient satisfaction. The between-group differences were analysed using a mixed-effects model for repeated measurements with a between group factor (I-group vs. C-group) and a time factor (baseline, 4 and 9 months). Ninety-eight patients were randomized (I-group: n = 48, C-group: n = 50), with 40 and 39 patients completing the study, respectively. Intention-to-treat analyses showed that both groups experienced statistically significant reductions in headache days/last month (I-group: -4 ± 6 days (95% CI 2.47; 5.95), p < .001) versus C-group: -4 ± 9 days ([95% CI 1.53; 6.79], p = .003), but there were no significant differences between groups (mean ± SE):Δ: 0.7 days ([95% CI, -2.50; 3.93], p = .66). At follow-up, 85% from the I-group and 86% from C-group, no longer fulfilled the criteria for MOH. The compliance rate was high, indicating that patients were motivated for receiving education, but we found no additional benefits of adding an educational programme to standard treatment. Future research focusing on the MOH complexity, group heterogeneity, duration and content of educational programmes is warranted. Randomized controlled trials (RCTs) of non-pharmacological intervention such as patient educational programmes are of great importance, as this approach is common in the clinical practice. Medication-overuse headache (MOH) is a heterogenetic patient group, which must be taken into account when conducting RCTs of non-pharmacological interventions. An educational programme based on Motivational Interviewing is well-tolerated among MOH patients, however, no superior effects were found from adding the educational programme to standard treatment versus standard treatment alone.


A gene required for class II-restricted antigen presentation maps to the major histocompatibility complex.
We have previously described a set of mutants (16.23-selected mutants) of a B lymphoblastoid cell line that are defective in the presentation of intact proteins to class II-restricted T cells, but effectively present immunogenic peptides. The mutations in these mutants are recessive in somatic cell hybrids and are not in Class II structural genes. Here, we report on a unique mutant, 5.2.4, in which a similar defect in class II-restricted antigen presentation has occurred in association with a one-megabase homozygous deletion in the class II region of the major histocompatibility complex (MHC). The defects in class II presentation among three of the 16.23-selected mutants, and between these mutants and 5.2.4, are noncomplementary in somatic cell hybrids. This suggests that the class II presentation-defective phenotype in all four mutants results from lesions in a single MHC-linked gene, a conclusion strengthened by the finding that in a hybrid made with a second, unrelated MHC deletion mutant, T2, the class II presentation defect in a 16.23-selected mutant is also not complemented. Mutant 5.2.4, in addition to its class II presentation defect, is also defective in surface expression of MHC class I molecules, most likely because its deletion encompasses the peptide supply factor 1 gene, whose function is known to be required for normal abundance of cell surface class I molecules. However, the surface abundance of class I molecules is normal in the 16.23-selected mutants, suggesting that the lesions affecting class I surface abundance and class II presentation result from mutations in different genes.


Modeling Cardiovascular Risks of E-Cigarettes With Human-Induced Pluripotent Stem Cell-Derived Endothelial Cells.
Electronic cigarettes (e-cigarettes) have experienced a tremendous increase in use. Unlike cigarette smoking, the effects of e-cigarettes and their constituents on mediating vascular health remain understudied. However, given their increasing popularity, it is imperative to evaluate the health risks of e-cigarettes, including the effects of their ingredients, especially nicotine and flavorings. The purpose of this study was to investigate the effects of flavored e-cigarette liquids (e-liquids) and serum isolated from e-cigarette users on endothelial health and endothelial cell-dependent macrophage activation. Human-induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) and a high-throughput screening approach were used to assess endothelial integrity following exposure to 6 different e-liquids with varying nicotine concentrations and to serum from e-cigarette users. The cytotoxicity of the e-liquids varied considerably, with the cinnamon-flavored product being most potent and leading to significantly decreased cell viability, increased reactive oxygen species (ROS) levels, caspase 3/7 activity, and low-density lipoprotein uptake, activation of oxidative stress-related pathway, and impaired tube formation and migration, confirming endothelial dysfunction. Upon exposure of ECs to e-liquid, conditioned media induced macrophage polarization into a pro-inflammatory state, eliciting the production of interleukin-1β and -6, leading to increased ROS. After exposure of human iPSC-ECs to serum of e-cigarette users, increased ROS linked to endothelial dysfunction was observed, as indicated by impaired pro-angiogenic properties. There was also an observed increase in inflammatory cytokine expression in the serum of e-cigarette users. Acute exposure to flavored e-liquids or e-cigarette use exacerbates endothelial dysfunction, which often precedes cardiovascular diseases.


Effect of dietary treatment on the plasma levels of lipids, lipoprotein cholesterol and LDL B protein in children with type II hyperlipoproteinemia.
These data confirm the effectiveness of therapeutic diets in children with type II hyperlipoproteinemia in conventional terms. However, the decrease in LDL cholesterol was not accompanied by a significant decrease in LDL B (suggesting only a small change in the number of LDL particles in plasma), and a fall in HDL cholesterol occurred, with no significant improvement in the LDL to HDL cholesterol ratio. These findings indicate that further research is needed to develop diets with more optimal effects. Finally, our hypothesis that such diets may produce an LDL particle of altered composition, needs to be confirmed using additional physical-chemical measurements of the size, composition and molecular weight distribution of LDL in plasma before and after dietary intervention.


Presynaptic Active Zone Density during Development and Synaptic Plasticity.
Neural circuits transmit information through synapses, and the efficiency of synaptic transmission is closely related to the density of presynaptic active zones, where synaptic vesicles are released. The goal of this review is to highlight recent insights into the molecular mechanisms that control the number of active zones per presynaptic terminal (active zone density) during developmental and stimulus-dependent changes in synaptic efficacy. At the neuromuscular junctions (NMJs), the active zone density is preserved across species, remains constant during development, and is the same between synapses with different activities. However, the NMJ active zones are not always stable, as exemplified by the change in active zone density during acute experimental manipulation or as a result of aging. Therefore, a mechanism must exist to maintain its density. In the central nervous system (CNS), active zones have restricted maximal size, exist in multiple numbers in larger presynaptic terminals, and maintain a constant density during development. These findings suggest that active zone density in the CNS is also controlled. However, in contrast to the NMJ, active zone density in the CNS can also be increased, as observed in hippocampal synapses in response to synaptic plasticity. Although the numbers of known active zone proteins and protein interactions have increased, less is known about the mechanism that controls the number or spacing of active zones. The following molecules are known to control active zone density and will be discussed herein: extracellular matrix laminins and voltage-dependent calcium channels, amyloid precursor proteins, the small GTPase Rab3, an endocytosis mechanism including synaptojanin, cytoskeleton protein spectrins and β-adducin, and a presynaptic web including spectrins. The molecular mechanisms that organize the active zone density are just beginning to be elucidated.


The N6-methyladenosine demethylase FTO is required for odontoblast differentiation in vitro and dentine formation in mice by promoting RUNX2 exon 5 inclusion through RBM4.
Fat mass and obesity-associated (FTO) protein, the first discovered N6-methyladenine (m6A) demethylase, played positive roles in bone formation. In this study, the aim was to investigate the function and potential mechanism of Fto in dentine formation. In vivo model, postnatal 12-day (PN12), 4-week-old (4 wk), 6-week-old (6 wk) healthy male C57BL/6J were randomly divided into Fto knockout (Fto-/- ) mice and wild-type (WT) littermates according to their genotypes, with 3-5 mice in each group. The mandibles of Fto-/- mice and WT control littermates were isolated for analysis by micro-computed tomography (micro-CT), 3-dimensional reconstruction and Haematoxylin-eosin (HE) staining. In vitro, mouse dental papilla cells (mDPCs) and human dental stem pulp cells (hDPSCs) were cultured with odontogenetic medium to evaluate differentiation capacity; expression levels of odontoblastic related genes were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). The inclusion levels of Runt-related transcription factor 2 (RUNX2) exon 5 in mDPCs and hDPSCs were detected by semiquantitative real-time polymerase chain reaction (RT-PCR). The RNA binding motif protein 4 (RBM4) m6A site was verified through m6A methylated RNA immunoprecipitation (MeRIP) and the stability of RBM4 mRNA influenced by FTO knockdown was measured by mRNA stability assay. Differences with p values < .05 were regarded as statistically significant. We discovered that Fto-/- mice showed significant dentine formation defects characterized by widened pulp cavity, enlarged pulp-tooth volume ratio, thinned dentine and pre-dentine layer of root (p < .05). Fto-/- mDPCs and FTO-silencing hDPSCs not only exhibited insufficient mineralization ability and decreased expression levels of odontoblastic mineralization related genes (p < .05), but showed significantly reduced Runx2 exon 5 inclusion level (p < .05). FTO knockdown increased the m6A level of RBM4 and destabilized the mRNA of RBM4, thus contributing to the reduced RBM4 expression level. Moreover, Rbm4 overexpression in Fto-/- mDPCs can partly restore Runx2 exon 5 inclusion level and the differentiation ability disrupted by Fto knockout. Thus, within the limitations of this study, the data suggest that FTO promotes odontoblastic differentiation during dentine formation by stabilizing RBM4 mRNA to promote RUNX2 exon 5 inclusion.


Opioid receptors: drivers to addiction?
Drug addiction is a worldwide societal problem and public health burden, and results from recreational drug use that develops into a complex brain disorder. The opioid system, one of the first discovered neuropeptide systems in the history of neuroscience, is central to addiction. Recently, opioid receptors have been propelled back on stage by the rising opioid epidemics, revolutions in G protein-coupled receptor research and fascinating developments in basic neuroscience. This Review discusses rapidly advancing research into the role of opioid receptors in addiction, and addresses the key questions of whether we can kill pain without addiction using mu-opioid-receptor-targeting opiates, how mu- and kappa-opioid receptors operate within the neurocircuitry of addiction and whether we can bridge human and animal opioid research in the field of drug abuse.


Comparative Analysis of Amino Acid Profiles in Patients with Glioblastoma and Meningioma Using Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (LC-ESI-MS/MS).
Brain tumors account for 1% of all cancers diagnosed de novo. Due to the specificity of the anatomical area in which they grow, they can cause significant neurological disorders and lead to poor functional status and disability. Regardless of the results of biochemical markers of intracranial neoplasms, they are currently of no diagnostic significance. The aim of the study was to use LC-ESI-MS/MS in conjunction with multivariate statistical analyses to examine changes in amino acid metabolic profiles between patients with glioblastoma, meningioma, and a group of patients treated for osteoarthritis of the spine as a control group. Comparative analysis of amino acids between patients with glioblastoma, meningioma, and the control group allowed for the identification of statistically significant differences in the amino acid profile, including both exogenous and endogenous amino acids. The amino acids that showed statistically significant differences (lysine, histidine, α-aminoadipic acid, phenylalanine) were evaluated for diagnostic usefulness based on the ROC curve. The best results were obtained for phenylalanine. Classification trees were used to build a model allowing for the correct classification of patients into the study group (patients with glioblastoma multiforme) and the control group, in which cysteine turned out to be the most important amino acid in the decision-making algorithm. Our results indicate amino acids that may prove valuable, used alone or in combination, toward improving the diagnosis of patients with glioma and meningioma. To better assess the potential utility of these markers, their performance requires further validation in a larger cohort of samples.


The postnatal development of post-activation potentiation in the rat neocortex.
The postnatal development of short-term potentiation (STP) and long-term potentiation (LTP) of the transcallosal response (TCR) was examined in anesthetized rats, aged 7 days (PN7) to adult (greater than or equal to PN180). Stimulation of callosal fibers produced a biphasic, positive-negative TCR recorded at the surface of the anterior neocortex in all age groups. The TCR showed a decrease in threshold, latency and halfwidth, and an increase in peak amplitude with age. High frequency callosal stimulation produced marked STP and LTP of the mature TCR. STP in young animals, however, could not be reliably detected until after PN16. LTP was not detected until PN18. STP and LTP approached adult levels rapidly after their initial appearance. The correlations between STP/LTP onset and development and known structural developments such as initial synaptogenesis, spine formation and myelinogenesis were not particularly strong. The implications of these results for specific hypotheses of STP/LTP mechanisms were discussed.


Optical and structural properties of nanostructured CuIn0.7Ga0.3(Se(1-x)Te(x))2 chalcopyrite thin films--effect of stoichiometry and annealing.
The aim of this work was to study the dependence of the optical, structural and morphological properties of CuIn0.7Ga0.3(Se(1-x)Te(x))2 (briefly CIGSeTe) thin films for two different stoichiometries (for x = 0.2 and 0.8). The films have been deposited onto soda lime glass (SLG) substrates by the e-beam evaporation technique. The films showed high absorption and revealed optical band gaps ranging from 1.17 eV to 1.06 eV for x = 0 with highest annealing temperatute at 525 degrees C and 1.12 eV to 1.02 eV for x = 0.8 and with highest annealed temperature at 600 degrees C. These results were correlated with the microstructural analysis by Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM) and X-ray diffractometry (XRD). The linear dependence of the lattice parameters as a function of Se and Te contents was examined. X-ray diffraction analyses showed that the films had the single phase chalcopyrite structure. The lattice parameters (a and c) varied linearly with the increase in Te content x from x = 0.2 to x = 0.8. The peak correspondng to the (1 1 2) plane orientation of the films increased with annealing process. Also, SEM images showed that both the grains size and the RMS (root mean square) values increased with annealing and higher Te amount that caused grains aggregation. The relative 600 degrees C elemental composition present in the deposited CIGS films have been measured by using energy dispersive X-ray analysis (EDX).


KIR Variation in Iranians Combines High Haplotype and Allotype Diversity With an Abundance of Functional Inhibitory Receptors.
Natural killer (NK) cells are innate lymphocytes that eliminate infected and transformed cells. They discriminate healthy from diseased tissue through killer cell Ig-like receptor (KIR) recognition of HLA class I ligands. Directly impacting NK cell function, KIR polymorphism associates with infection control and multiple autoimmune and pregnancy syndromes. Here we analyze KIR diversity of 241 individuals from five groups of Iranians. These five populations represent Baloch, Kurd, and Lur, together comprising 15% of the ethnically diverse Iranian population. We identified 159 KIR alleles, including 11 not previously characterized. We also identified 170 centromeric and 94 telomeric haplotypes, and 15 different KIR haplotypes carrying either a deletion or duplication encompassing one or more complete KIR genes. As expected, comparing our data with those representing major worldwide populations revealed the greatest similarity between Iranians and Europeans. Despite this similarity we observed higher frequencies of KIR3DL1*001 in Iran than any other population, and the highest frequency of HLA-B*51, a Bw4-containing allotype that acts as a strong educator of KIR3DL1*001+ NK cells. Compared to Europeans, the Iranians we studied also have a reduced frequency of 3DL1*004, which encodes an allotype that is not expressed at the NK cell surface. Concurrent with the resulting high frequency of strong viable interactions between inhibitory KIR and polymorphic HLA class I, the majority of KIR-A haplotypes characterized do not express a functional activating receptor. By contrast, the most frequent KIR-B haplotype in Iran expresses only one functional inhibitory KIR and the maximum number of activating KIR. This first complete, high-resolution, characterization of the KIR locus of Iranians will form a valuable reference for future clinical and population studies.


Spin-lattice relaxation in murine tumors after in vivo treatment with interferon alpha/beta or tumor necrosis factor alpha.
1H NMR spin-lattice relaxation times (T1) were measured in vitro and in vivo in Friend leukemia cell tumors during subcutaneous tumor growth in syngeneic mice and after in vivo administration of either purified murine interferon alpha/beta (IFN) or recombinant tumor necrosis factor alpha (TNF). Untreated tumors exhibited monoexponential T1 relaxation independently of tumor age at least until Day 16 after implantation. Histological examinations showed that under these conditions tumors were highly homogeneous and substantially free of necrotic areas. Peritumoral administrations of either IFN or TNF did not significantly alter the tumor relaxation properties at early stages of inhibition of tumor growth. The longitudinal relaxation decay became instead clearly biexponential at later stages (more than 7 days of IFN treatment or 2 days after TNF administration). While the T1 relaxation behavior could be unequivocally correlated with the presence of necrotic areas in these tumors, it could not be considered as an early marker of the altered growth capability, induced by administration of either IFN or TNF.


G protein-coupled receptor desensitization as a measure of signaling: modeling of arrestin recruitment to activated CCK-B receptors.
Gastrin is one of the principle hormonal mediators of gastric acid secretion, and its cognate receptor (CCK-B) is a member of the superfamily of GPCRs. Patients with hypergastrinemia may present with a variety of symptoms, including gastric ulcers or malignant tumors. Thus, the molecular mechanisms that terminate CCK-B receptor signaling, as well as an ability to measure gastrin bioactivity in a timely manner, have important clinical implications. In order to assess CCK-B receptor regulation, we have constructed a single cell biosensor containing the CCK-B receptor and an arrestin/GFP chimera. The gastrin biosensor responded to both immunologically detectable gastrin-17 and undetectable pentagastrin, and was able to determine the gastrin bioactivity of serum from a patient with clinical hypergastrinemia. We determined that the CCK-B receptor binds arrestin with a pharmacology mirroring CCK-B receptor signaling through inositol phosphate, and that the rate of arrestin dissociation from internalized receptor mirrors receptor recycling to the plasma membrane. Moreover, the CCK-B recycling rate is intermediate between that of Class A GPCRs such as the beta2-adrenergic receptor and Class B GPCRs such as the vasopressin type 2 receptor. Mathematical modeling of these results indicates that a common receptor conformation may underlie both CCK-B signaling and desensitization. In addition to its use in drug screening, this methodology should generalize to other receptors for use in diagnosis and monitoring of bioactive ligands involved in GPCR-based disease.


Hoxb13, a potential prognostic biomarker for prostate cancer.
HOXB13, a member of the homeobox proteins family, is a key regulator of the epithelial differentiation in the prostate gland. HOXB13 is overexpressed during malignant progression of the prostatic tissue and suspected to contribute in the pathogenesis of the prostate gland. In androgen deprived conditions, HOXB13 is thought to act through inhibition of the tumour suppressor protein p21. Since HOXB13 has a multifaceted role in ventral prostate development, its critical partners in the cascade need to be elucidated for a further understanding of its role in prostate malignancy. In this report, we review the functions attributed to HOXB13, by highlighting the most recent findings supporting the hypothesis that HOXB13 might serve as a novel biomarker for the prognosis of prostate cancer.


Visually storying living with HIV: bridging stressors and supports in accessing care.
This paper examines how visual narratives may bridge relational understandings between people living with HIV/AIDS (PLWH/A) and future oral health care providers. Borrowing from literature in participatory visual methods such as photo elicitation and photovoice, we explored how PLWH/A visually choose to represent their daily lives. This study uses a grounded theory action-oriented approach in examining the thematic analysis of 257 photos and 12 related reflective participant journals. Ten collaborative themes emerged from the participants' analysis of their photos. These themes of social support, places, family, staff, group, recovery tools, transportation, friends, medications and food exhibited the indivisible characteristics of stressors and supports commonly found in accessing care. Further researcher reflections also found three meta-themes of stigmatization, maintenance of positive mental health and the development of pride in managing one's health. PLWH/A need to share these visual themes of supports and stressors with future dental providers so that they may hopefully acquire an understanding of chronic illness that is more personalized and relationship centered rather than merely numeric and detached.


KCNQ4 mutations associated with nonsyndromic progressive sensorineural hearing loss.
This article provides an update on the current progress in identification of KCNQ4 mutations responsible for DFNA2, a subtype of autosomal dominant nonsyndromic progressive hearing loss. Hearing loss in pateints with DFNA2 usually start at high frequencies in their 20s and 30s, and then progress to more than 60 dB in less than 10 years, with middle and low frequencies often affected as well. To date, eight missense mutations and two deletions of the KCNQ4 gene have been identified in patients with DFNA2 with various clinical phenotypes. In general, missense mutations are associated with younger-onset and all-frequency hearing loss, whereas deletion mutations are underlying later-onset and pure high-frequency hearing loss. The etiology of DFNA2 remains largely unknown at this point, even though the degeneration of cochlear outer hair cells, caused by dysfunction of KCNQ4 channels, might be one of the underlying mechanisms. During the last decade, significant progress has been made in identifying KCNQ4 mutations in patients with DFNA2. Elucidation of the pathogenic effect of these mutations will help to gain insights into the molecular mechanisms of hearing and hearing loss, which, in turn, will facilitate informative genetic counseling, early diagnosis, and even treatment of hearing loss.


Modulating secretory pathway pH by proton channel co-expression can increase recombinant protein stability in plants.
Eukaryotic expression systems are used for the production of complex secreted proteins. However, recombinant proteins face considerable biochemical challenges along the secretory pathway, including proteolysis and pH variation between organelles. As the use of synthetic biology matures into solutions for protein production, various host-cell engineering approaches are being developed to ameliorate host-cell factors that can limit recombinant protein quality and yield. We report the potential of the influenza M2 ion channel as a novel tool to neutralize the pH in acidic subcellular compartments. Using transient expression in the plant host, Nicotiana benthamiana, we show that ion channel expression can significantly raise pH in the Golgi apparatus and that this can have a strong stabilizing effect on a fusion protein separated by an acid-susceptible linker peptide. We exemplify the utility of this effect in recombinant protein production using influenza hemagglutinin subtypes differentially stable at low pH; the expression of hemagglutinins prone to conformational change in mildly acidic conditions is considerably enhanced by M2 co-expression. The co-expression of a heterologous ion channel to stabilize acid-labile proteins and peptides represents a novel approach to increasing the yield and quality of secreted recombinant proteins in plants and, possibly, in other eukaryotic expression hosts.


The RING-Type E3 Ligase XBAT35.2 Is Involved in Cell Death Induction and Pathogen Response.
XBAT35 belongs to a subfamily of Arabidopsis (Arabidopsis thaliana) RING-type E3s that are similar in domain architecture to the rice (Oryza sativa) XA21 Binding Protein3, a defense protein. The XBAT35 transcript undergoes alternative splicing to produce two protein isoforms, XBAT35.1 and XBAT35.2. Here, we demonstrate that XBAT35.2 localizes predominantly to the Golgi and is involved in cell death induction and pathogen response. XBAT35.2, but not XBAT35.1, was found to trigger cell death when overexpressed in tobacco (Nicotiana benthamiana) leaves and does so in a manner that requires its RING domain. Loss of XBAT35 gene function disrupts the plant's ability to defend against pathogen attack, whereas overexpression of XBAT35.2 enhances resistance to pathogens. XBAT35.2 was found to be unstable and promotes its own degradation, suggesting self-regulation. Inoculation with virulent and avirulent strains of the bacterial pathogen Pseudomonas syringae pv tomato DC3000 results in a drastic reduction in the levels of ubiquitinated XBAT35.2 and an increase in the abundance of the E3. This implies that pathogen infection prohibits XBAT35.2 self-regulation and stabilizes the E3. In agreement with a role in defending against pathogens, XBAT35.2 interacts with defense-related Accelerated Cell Death11 (ACD11) in planta and promotes the proteasome-dependent turnover of ACD11 in cell-free degradation assays. In accordance with regulation by a stabilized XBAT35.2, the levels of ubiquitinated ACD11 increased considerably, and the abundance of ACD11 was reduced following pathogen infection. In addition, treatment of transgenic seedlings with a proteasome inhibitor results in the accumulation of ACD11, confirming proteasome-dependent degradation. Collectively, these results highlight a novel role for XBAT35.2 in cell death induction and defense against pathogens.


[Hemomicrocirculatory disorders in pathogenesis of entero-enteroanastomosis insufficiency].
Morphofunctional changes of hemomicrocirculation in the walls of the small intestine in acute strangulation intestinal obstruction and after its surgical elimination were studied experimentally on 60 mongrel dogs (weight 7-20 kg). It is demonstrated that main causes of complications in postoperative period were disorders of microcirculation in the walls of the small intestine and inflammation in the bringing part of the small intestine. On day 3 to 7 after elimination of strangulation intestinal obstruction hemomicrocirculation in tissues of bringing loop of small intestine did not meet the requirements for reparation after resection of necrotic part and creation of entero-enteroanastomosis. This is the cause of insufficiency of entero-enteroanastomosis in this time interval.


[Various luminescence characteristics of allocytoplasmic wheat hybrids].
Comparative studies of thermoluminescence and slow fluorescence induction were carried out on leaves of allocytoplasmic hybrids and standard regionalized wheat (cultivar Zarya). There are differences in luminescence indices between these objects correlated to the thermoluminescence and slow fluorescence induction methods. These differences allow estimation of the functional activity of the photosynthetic system. All studied allocytoplasmic hybrids exceeded the standard wheat in both luminescence indices FM/FT (slow fluorescence induction) and SA/Stot (thermoluminescence) and productivity (height of seedlings, number of ears and grains, grain mass, etc.). The highest values of FM/FT and SA/Stot and the lowest value of SC/Stot were recorded in the allocytoplasmic hybrid Igen 3 x Mironovskaya 808, which is characterized by the highest productivity.


Dietary supplementation with long-chain monounsaturated fatty acids attenuates obesity-related metabolic dysfunction and increases expression of PPAR gamma in adipose tissue in type 2 diabetic KK-Ay mice.
The objective of present study was to examine the effect of long-chain monounsaturated fatty acids (LC-MUFAs) with chain lengths longer than 18 (i.e., C20:1 and C22:1 isomers combined) on obesity-related metabolic dysfunction and its molecular mechanisms. Type-2 diabetic KK-Ay mice (n = 20) were randomly assigned to the 7% soybean oil-diet group (control group) and 4% LC-MUFA concentrate-supplemented-diet group (LC-MUFA group). At 8 weeks on the diet, the results showed that plasma, liver and adipose tissue levels of C20:1 and C22:1 isomers increased significantly with LC-MUFA treatment. Supplementation with LC-MUFAs markedly reduced white fat pad weight as well as adipocyte size in the mice. The levels of plasma free fatty acids, insulin, and leptin concentration in the obese diabetic mice of the LC-MUFA group were also decreased as compared with the mice in the soybean oil-diet control group. Dietary LC-MUFAs significantly increased the mRNA expression of peroxisome proliferator-activated receptor gamma (Pparg), lipoprotein lipase (Lpl), fatty acid transport protein (Fatp), fatty acid translocase/CD36 (Cd36), as well as mRNA expression of genes involved in lipid oxidation such as carnitine palmitoyltransferase-1A (Cpt1a) and citrate synthase (Cs), and decreased the mRNA expression of inflammatory marker serum amyloid A 3 (Saa3) in the adipose tissues of diabetic mice. The results suggest that LC-MUFAs may ameliorate obesity-related metabolic dysfunction partly through increased expression of Pparg as well as its target genes, and decreased inflammatory marker expression in white adipose tissue.


Noninvasive cardiovascular phenotyping in mice.
With the growth of genetic engineering, mice have become common as models of human diseases, which in turn has stimulated the development of techniques to monitor and image the murine cardiovascular system. Invasive methods are often more quantitative, but noninvasive methods are preferred when measurements must be repeated serially on living animals during development or in response to pharmacological or surgical interventions. Because of the small size and high heart rates in mice, high spatial and temporal resolutions are required to preserve signal fidelity. Monitoring of body temperature and the electrocardiogram is essential when animals must be anesthetized for a measurement or other procedure. Several other groups have developed cardiovascular imaging modalities suitable for murine applications, and ultrasound is the most widely used. Our group has developed and applied high-resolution Doppler probes and signal processing for measuring blood velocity in the heart and peripheral vessels of anesthetized mice noninvasively. We can measure cardiac filling and ejection velocities as indices of systolic and diastolic ventricular function and for timing of cardiac events; velocity pulse arrival times for determining pulse-wave velocity and arterial stiffness; peripheral velocity waveforms as indices of arterial resistance, compliance, and wave reflections; stenotic velocities for estimation of pressure drop and detection of vorticity; and tail artery velocity for determining systolic and diastolic blood pressure using a pressure cuff. These noninvasive methods are convenient and easy to apply and have been used to detect and evaluate numerous cardiovascular phenotypes in mutant mice.


Connect the dots: sketching out microbiome interactions through networking approaches.
Microbiome networking analysis has emerged as a powerful tool for studying the complex interactions among microorganisms in various ecological niches, including the human body and several environments. This analysis has been used extensively in both human and environmental studies, revealing key taxa and functional units peculiar to the ecosystem considered. In particular, it has been mainly used to investigate the effects of environmental stressors, such as pollution, climate change or therapies, on host-associated microbial communities and ecosystem function. In this review, we discuss the latest advances in microbiome networking analysis, including methods for constructing and analyzing microbiome networks, and provide a case study on how to use these tools. These analyses typically involve constructing a network that represents interactions among microbial taxa or functional units, such as genes or metabolic pathways. Such networks can be based on a variety of data sources, including 16S rRNA sequencing, metagenomic sequencing, and metabolomics data. Once constructed, these networks can be analyzed to identify key nodes or modules important for the stability and function of the microbiome. By providing insights into essential ecological features of microbial communities, microbiome networking analysis has the potential to transform our understanding of the microbial world and its impact on human health and the environment.


Diagnostic test accuracy of informant-based tools to diagnose dementia in older hospital patients with delirium: a prospective cohort study.
delirium and dementia co-exist commonly in hospital. Older people with delirium have high rates of undiagnosed dementia, but delirium affects the use of cognitive testing in dementia diagnosis. Novel methods to detect dementia in delirium are needed. The purpose of the study was to investigate the diagnostic test accuracy of informant tools to detect dementia in hospitalised older people with delirium. the presence of dementia on admission was assessed using the short form of the Informant Questionnaire of Cognitive Decline in the Elderly (IQCODE-SF) and Alzheimer's Disease 8 (AD8) in people over 70 years old with delirium. Reference standard diagnosis was made using Diagnostic and Statistical Manual of Mental Disorders fourth edition (DSM-IV) criteria at 3 months. The main outcome measures were the diagnostic test accuracy of the IQCODE-SF and the AD8 in diagnosing DSM-IV dementia. dementia prevalence at 3 months was 61%. The area under the receiver operating characteristic curve (AUROC) was 0.93 (P < 0.0005) for admission IQCODE-SF and 0.91 (P < 0.0005) for admission AD8. An IQCODE-SF test result of >3.82 on admission had a sensitivity of 0.91 (0.79-0.97) and specificity of 0.93 (0.76-0.99) for detecting dementia. An AD8 of >6 had a sensitivity of 0.83 (0.69-0.92) and specificity of 0.90 (0.72-0.97) for detecting dementia. the IQCODE-SF and AD8 are sensitive and specific tools to detect prior dementia in older people with delirium. The routine use of either tool in practice could have important clinical impact, by improving the recognition and hence management of those with dementia.


Seven steps to assure the best possible treatment of persons with glaucoma or suspected of having glaucoma.
Glaucoma causes more blindness and disability than any other eye disease except for cataract. Cataract surgery is highly successful, even intracapsular cataract surgery. The major problem, then, with regard to relieving or preventing disability related to cataract is one of access to care. Though access to care is a major problem with glaucoma, it is by no means the only problem. Many patients with glaucoma get worse, even when they are being treated. Additionally, it is not only glaucoma itself that causes disability. In the recent past, far more damage was caused by treatment for glaucoma than by glaucoma itself. To repeat, summarized slightly differently, glaucoma and its treatment damage health more than any other eye disease except cataract, and the medical profession is responsible for a major part of that problem. How can it be that glaucoma and its treatment still cause so much disability? In the first place, care is unavailable in many areas, and, second, care is not utilized even where available, and third, care is often inappropriate even where available. The first two issues are of great importance but are not the subject of this discussion. It is my belief that the major explanation for why care is often poor is because the goals of care are often wrong. Specifically, the goals of care are to lower intraocular pressure, or to prevent visual field loss. These may be appropriate intermediary steps on the way to achieving the appropriate goal, but they are not the goal itself.


National surveillance of influenza-associated encephalopathy in Japan over six years, before and during the 2009-2010 influenza pandemic.
Influenza-associated encephalopathy (IAE) is a serious complication of influenza and is reported most frequently in Japan. This paper presents an assessment of the epidemiological characteristics of influenza A (H1N1) 2009-associated encephalopathy in comparison to seasonal IAE, based on Japanese national surveillance data of influenza-like illness (ILI) and IAE during flu seasons from 2004-2005 through 2009-2010. In each season before the pandemic, 34-55 IAE cases (mean 47.8; 95% confidence interval: 36.1-59.4) were reported, and these cases increased drastically to 331 during the 2009 pandemic (6.9-fold the previous seasons). Of the 331 IAE cases, 322 cases were reported as influenza A (H1N1) 2009-associated encephalopathy. The peaks of IAE were consistent with the peaks of the influenza epidemics and pandemics. A total of 570 cases of IAE (seasonal A, 170; seasonal B, 50; influenza A (H1N1) 2009, 322; unknown, 28) were reported over six seasons. The case fatality rate (CFR) ranged from 4.8 to 18.2% before the pandemic seasons and 3.6% in the 2009 pandemic season. The CFR of pandemic-IAE was 3.7%, which is lower than that of influenza A-/B-associated encephalopathy (12.9%, p<0.001; 14.0%, p = 0.002; respectively). The median age of IAE was 7 years during the pandemic, which is higher than that of influenza A-/B-associated encephalopathy (4, p<0.001; 4.5, p = 0.006; respectively). However, the number of pandemic-IAE cases per estimated ILI outpatients peaked in the 0-4-year age group and data both before and during the pandemic season showed a U-shape pattern. This suggests that the high incidence of influenza infection in the 0-4 year age group may lead to a high incidence of IAE in the same age group in a future influenza season. Further studies should include epidemiologic case definitions and clinical details of IAE to gain a more accurate understanding of the epidemiologic status of IAE.


Exome sequencing-aided precise diagnosis of four families with type I Stickler syndrome.
Stickler syndrome is a multisystemic disorder characterized by ophthalmological and non-ophthalmological abnormalities, frequently misdiagnosed due to high clinical heterogeneity. Stickler syndrome type I (STL1) is predominantly caused by mutations in the COL2A1 gene. Exome sequencing and co-segregation analysis were utilized to scrutinize 35 families with high myopia, and pathogenic mutations were identified. Mutant COL2A1 was overexpressed in cells for mechanistic study. A retrospective genotype-phenotype correlation analysis was further conducted. Two novel pathogenic mutations (c.2895+1G>C and c.3505G>A (p.Val1169Ile)) and two reported mutations (c.1597C>T (p.Arg533*) and c.1693C>T (p.Arg565Cys)) in COL2A1 were identified causing STL1. These mutations are all in the G-X-Y triplet, and c.2895+1G>C contributed to aberrant RNA splicing. COL2A1 mutants tended to form large aggregates in the endoplasmic reticulum (ER) and elevated ER stress. Additionally, mutations c.550G>A (p.Ala184Thr) and c.2806G>A (p.Gly936Ser) in COL2A1 were found in high myopia families, but were likely benign, although c.2806G>A (p.Gly936Ser) is on G-X-Y triplet. Moreover, genotype-phenotype correlation analysis revealed that mutations in exon 2 mainly contribute to retinal detachment, whereas mutations in the collagen alpha-1 chain region of COL2A1 tend to cause non-ophthalmologic symptoms. This study broadens the COL2A1 gene mutation spectrum, provides evidence for ER stress caused by pathogenic COL2A1 mutations and highlights the importance of non-ophthalmological examination in clinical diagnosis of high myopia.


Factors influencing self-management among Indonesian patients with early-stage chronic kidney disease: A cross-sectional study.
The purpose of this study was to explore the self-management of patients with early-stage chronic kidney disease (CKD) and its influencing factors. A convenient sample of 226 patients with early-stage CKD was recruited from 63 Public Health Centers in Indonesia, from June to September 2020. Demographic characteristics, health literacy, illness perception, self-efficacy and self-management were assessed using self-reported questionnaires. Stepwise multiple linear regression analysis was performed to identify the factors influencing self-management. This study adhered to the EQUATOR checklist, STROBE. The mean estimated glomerular filtration rate was 63.45 ml/min/1.73 m2 (standard deviation [SD] = 15.34). The average scores for health literacy, illness perception, self-efficacy and self-management were 32.11 (SD = 4.46), 4.57 (SD = 1.46), 183.64 (SD = 38.23) and 76.92 (SD = 9.45), respectively. The influencing factors were education level, monthly income, family history of comorbidity, health literacy and self-efficacy, which accounted for 45% of total self-management score. Indonesian patients with early-stage CKD showed low level of health literacy, but positive illness perception and self-efficacy; these factors significantly affected CKD self-management. Health literacy was found to influence all dimensions of self-management: self-integration, problem-solving, seeking social support and adherence to the recommended regimen. Adherence to the recommended regimen is the most challenging dimension of CKD self-management. Health literacy was found to be a major determinant of self-management. Improving health literacy and motivation of patients with early-stage CKD may help sustain positive illness perception and self-efficacy, and improve self-management.


A VicRK signal transduction system in Streptococcus mutans affects gtfBCD, gbpB, and ftf expression, biofilm formation, and genetic competence development.
Bacteria exposed to transient host environments can elicit adaptive responses by triggering the differential expression of genes via two-component signal transduction systems. This study describes the vicRK signal transduction system in Streptococcus mutans. A vicK (putative histidine kinase) deletion mutant (SmuvicK) was isolated. However, a vicR (putative response regulator) null mutation was apparently lethal, since the only transformants isolated after attempted mutagenesis overexpressed all three genes in the vicRKX operon (Smuvic+). Compared with the wild-type UA159 strain, both mutants formed aberrant biofilms. Moreover, the vicK mutant biofilm formed in sucrose-supplemented medium was easily detachable relative to that of the parent. The rate of total dextran formation by this mutant was remarkably reduced compared to the wild type, whereas it was increased in Smuvic+. Based on real-time PCR, Smuvic+ showed increased gtfBCD, gbpB, and ftf expression, while a recombinant VicR fusion protein was shown to bind the promoter regions of the gtfB, gtfC, and ftf genes. Also, transformation efficiency in the presence or absence of the S. mutans competence-stimulating peptide was altered for the vic mutants. In vivo studies conducted using SmuvicK in a specific-pathogen-free rat model resulted in significantly increased smooth-surface dental plaque (Pearson-Filon statistic [PF], <0.001). While the absence of vicK did not alter the incidence of caries, a significant reduction in SmuvicK CFU counts was observed in plaque samples relative to that of the parent (PF, <0.001). Taken together, these findings support involvement of the vicRK signal transduction system in regulating several important physiological processes in S. mutans.


Further characterization of conditional lethal amber nonsense mutants of vesicular stomatitis virus: nucleotide sequence analysis.
Conditional lethal amber nonsense mutants of vesicular stomatitis virus, Indiana serotype, classified in complementation group I (the L gene), synthesize truncated versions of the L protein. This paper reports further characterization of mutants AmbL1, AmbL2 and AmbL3 by nucleic acid sequence analysis, which was achieved by sequencing L mRNA directly using appropriate synthetic oligonucleotides. In each case a single point mutation altered a glutamine-specifying codon to an amber stop codon. The L mRNA from wild-type and revertant viruses was sequenced for comparison. Of the revertants sequenced, each had reverted by back mutation within the same codon as the original mutation. A revertant of AmbL2 reverted by a second site mutation, also within the same codon as the original mutation. These mutants may be useful for assigning functions to different parts of the L polypeptide chain.


A EAACI drug allergy interest group survey on how European allergy specialists deal with β-lactam allergy.
An accurate diagnosis of β-lactam (BL) allergy can reduce patient morbidity and mortality. Our aim was to investigate the availability of BL reagents, their use and test procedures in different parts of Europe, as well as any differences in the diagnostic workups for evaluating subjects with BL hypersensitivity. A survey was emailed to all members of the EAACI Drug Allergy Interest Group (DAIG) between February and April 2016, and the questionnaire was meant to study the management of suspected BL hypersensitivity. The questionnaire was emailed to 82 DAIG centres and answered by 57. Amoxicillin alone or combined to clavulanic acid were the most commonly involved BL except in the Danish centre, where penicillin V was the most frequently suspected BL. All centres performed an allergy workup in subjects with histories of hypersensitivity to BL: 53 centres (93%) followed DAIG guidelines, two national guidelines and two local guidelines. However, there were deviations from DAIG recommendations concerning allergy tests, especially drug provocation tests. A significant heterogeneity exists in current practice not only among countries, but also among centres within the same country. This suggests the need to re-evaluate, update and standardize protocols on the management of patients with suspected BL allergy.


Involvement in the Juvenile Justice System for African American Adolescents: Examining Associations with Behavioral Health Problems.
While researchers have found that African American youth experience higher levels of juvenile justice involvement at every system level (arrest, sentencing, and incarceration) relative to their other ethnic counterparts, few studies have explored how juvenile justice involvement and number of contacts might be correlated with this broad range of problems. A convenience sample of 638 African American adolescents living in predominantly low-income, urban communities participated in a survey related to juvenile justice involvement. Major findings using logistic regression models indicated that adolescents who reported juvenile justice system involvement versus no involvement were 2.3 times as likely to report mental health problems, substance abuse, and delinquent or youth offending behaviors. Additional findings documented that the higher the number of juvenile justice system contacts, the higher the rates of delinquent behaviors, alcohol and marijuana use, sex while high on drugs, and commercial sex. These findings suggest that identifying and targeting youth who have multiple juvenile justice system contacts, especially those in low-resourced communities for early intervention services, may be beneficial. Future research should examine whether peer network norms might mediate the relationships between juvenile justice involvement and youth problem behaviors.


Interrelationship between cardiac hypertrophy, heart failure, and chronic kidney disease: endoplasmic reticulum stress as a mediator of pathogenesis.
Synthesis of transmembrane and secretory proteins occurs within the endoplasmic reticulum (ER) and is extremely important in the normal functioning of both the heart and kidney. The dysregulation of protein synthesis/processing within the ER causes the accumulation of unfolded proteins, thereby leading to ER stress and the activation of the unfolded protein response. Sarcoplasmic reticulum/ER Ca2+ disequilibrium can lead to cardiac hypertrophy via cytosolic Ca2+ elevation and stimulation of the Ca2+/calmodulin, calcineurin, NF-AT3 pathway. Although cardiac hypertrophy may be initially adaptive, prolonged or severe ER stress resulting from the increased protein synthesis associated with cardiac hypertrophy can lead to apoptosis of cardiac myocytes and result in reduced cardiac output and chronic heart failure. The failing heart has a dramatic effect on renal function because of inadequate perfusion and stimulates the release of many neurohumoral factors that may lead to further ER stress within the heart, including angiotensin II and arginine-vasopressin. Renal failure attributable to proteinuria and uremia also induces ER stress within the kidney, which contributes to the transformation of tubular epithelial cells to a fibroblast-like phenotype, fibrosis, and tubular cell apoptosis, further diminishing renal function. As a consequence, cardiorenal syndrome may develop into a vicious circle with poor prognosis. New therapeutic modalities to alleviate ER stress through stimulation of the cytoprotective components of the unfolded protein response, including GRP78 upregulation and eukaryotic initiation factor 2α phosphorylation, may hold promise to reduce the high morbidity and mortality associated with cardiorenal syndrome.


Adenovirus-mediated gene therapy in an experimental model of breast cancer metastatic to the brain.
We investigated the therapeutic efficacy of adenovirus-mediated gene therapy to treat malignant mammary tumors in vitro and in vivo in the brain. A mammary adenocarcinoma cell line derived from Fischer rats (13762 MAT B III; MAT-B) was used. In vitro studies demonstrated that the MAT-B cells could be efficiently transduced with a replication-defective adenovirus (ADV) vector that carried the herpes simplex virus gene for thymidine kinase (ADV-tk), and that ADV-tk transduction rendered the MAT-B cells sensitive to killing, in a dose-dependent manner, with ganciclovir (GCV). An animal model of a mammary tumor metastatic to the brain was produced by injecting MAT-B cells into the caudate nucleus of Fischer rats. Seven days after MAT-B cell injection, when the tumors were approximately 5 mm2 in cross-sectional size, the tumors were injected with ADV-tk or a control adenovirus vector containing the beta-galactosidase (beta-Gal) gene (ADV-beta gal). After vector injection the animals were treated with GCV or with saline for 6 days. Sixteen days after tumor cell injection, the brains were examined histologically. The rats that were injected with ADV-beta gal and treated with GCV or saline, and those that were injected with ADV-tk and treated with saline had large tumors, whereas the rats that were injected with ADV-tk and treated with GCV had no visible tumor tissue at the site of tumor cell injection. In survival studies animals treated with ADV-tk+GCV survived a significantly longer time than animals treated with ADV-beta gal+GCV. Our results demonstrate that the recombinant adenoviral vector containing the tk gene confers GCV cytotoxic sensitivity to mammary tumor cells in vitro and in the brain, and suggest that this treatment strategy may be useful in treating somatic tumors that metastasize to the brain.


Expression of the Saccharomyces cerevisiae Kex2p endoprotease in inset cells. Evidence for a carboxy-terminal autoprocessing event.
The pheromone-processing Kex2p endoprotease of Saccharomyces cerevisiae has been difficult to characterize due to its low level of expression in yeast cells. To overcome this problem, we have overexpressed Kex2p using the baculovirus/insect cell expression system. Spodoptera frugiperda Sf9 insect cells infected with a recombinant baculovirus, containing the complete KEX2 gene which encodes the Kex2p protease (814 amino acids), accumulate an 120-kDa functional form of the enzyme. The inhibition profile of the insect-cell-derived endoprotease is similar to that of the yeast enzyme. The recombinant infected insect cells also secrete into the medium about half of the total Kex2p activity produced. Deleting the carboxyl-terminal tail and the transmembrane domain of Kex2p (Kex2 delta p, 666 amino acids) does not measurably interfere with the enzyme characteristics and results in the secretion of up to 90% of the total enzyme activity. The truncated form, Kex2 delta p, of the endoprotease accumulates in the cell supernatant to 6.7 x 10(5) U/l. The molecular mass of the secreted forms for both the wild-type Kex2p and Kex2 delta p is the same (70 kDa) and is 50-kDa lower than the intracellular form. This result implicates a processing event which gives rise to shorter extracellular forms of both the wild-type Kex2p and Kex2 delta p and which trims their carboxy termini upsteam of amino acid 666. This processing event requires the integrity of the Ser385 of the Kex2p active site.


Infrared and Near-Infrared Spectrometry of Anatase and Rutile Particles Bandgap Excited in Liquid.
Chemical conversion of materials is completed in milliseconds or seconds by assembling atoms over semiconductor photocatalysts. Bandgap-excited electrons and holes reactive on this time scale are key to efficient atom assembly to yield the desired products. In this study, attenuated total reflection of infrared and near-infrared light was applied to characterize and quantify the electronic absorption of TiO2 photocatalysts excited in liquid. Nanoparticles of rutile or anatase were placed on a diamond prism, covered with liquid, and irradiated by steady UV light through the prism. Electrons excited in rutile particles (JRC-TIO-6) formed small polarons characterized by a symmetric absorption band spread over 10000-700 cm-1 with a maximum at 6000 cm-1. Electrons in anatase particles (JRC-TIO-7) created large polarons and produced an asymmetric absorption band that gradually strengthened at wavenumbers below 5000 cm-1 and sharply weakened at 1000 cm-1. The absorption spectrum of large electron polarons in TIO-7 was compared with the absorption reported in a Sr-doped NaTaO3 photocatalyst, and it was suggested that excited electrons were accommodated as large polarons in NaTaO3 photocatalysts efficient for artificial photosynthesis. UV-light power dependence of the absorption bands was observed in N2-exposed decane liquid to deduce electron-hole recombination kinetics. With light power density P > 200 W m-2 (TIO-6) and 2000 W m-2 (TIO-7), the polaron absorptions were enhanced with absorbance being proportional to P1/2. The observed 1/2-order power law suggested recombination of multiple electrons and holes randomly moving in each particle. Upon excitation with smaller P, the power-law order increased to unity. The unity-order power law was interpreted with recombination of an electron and a hole that were excited by the same photon. In addition, an average lifetime of 1 ms was estimated with electron polarons in TIO-6 when weakly excited at P = 20 W m-2 to simulate solar-light irradiation.


Carbon monoxide and lethal arrhythmias.
The effect of acute exposure to carbon monoxide on ventricular arrhythmias was studied in a previously described chronically maintained animal model of sudden cardiac death. In 60 percent of dogs with a healed anterior myocardial infarction, the combination of mild exercise and acute myocardial ischemia induces ventricular fibrillation. The events in this model are highly reproducible, thus allowing study by internal control analysis. Dogs that develop ventricular fibrillation during the test of exercise and acute myocardial ischemia are considered at high risk for sudden death and are defined as "susceptible"; dogs that survive the test without a fatal arrhythmia are considered at low risk for sudden death and are defined as "resistant." In the current study, the effects of carboxyhemoglobin levels ranging from 5 to 15 percent were tested in resistant and susceptible dogs. A trend toward higher heart rates was observed at all levels of carboxyhemoglobin, although significant differences were observed only with 15 percent carboxyhemoglobin. This trend was observed at rest and during exercise in both resistant and susceptible dogs. In resistant animals, in which acute myocardial ischemia is typically associated with bradycardia even under the control condition, this reflex response occurred earlier and was augmented after exposure to carbon monoxide. This effect may depend on the increased hypoxic challenge caused by carbon monoxide, and thus on an augmentation of the neural reflex activation or a sensitization of the sinus node to acetylcholine induced by hypoxia. In both resistant and susceptible dogs, carbon monoxide exposure induced a worsening of ventricular arrhythmias in a minority of cases. This worsening was not reproducible in subsequent trials. These data indicate that acute exposure to carbon monoxide is seldom arrhythmogenic in dogs that have survived myocardial infarction. Nevertheless, the observation that carbon monoxide exposure increases heart rate at rest and during moderate exercise may have clinical implications relevant to patients with coronary artery disease.(ABSTRACT TRUNCATED AT 400 WORDS)


The lumenal domain of Sec63p stimulates the ATPase activity of BiP and mediates BiP recruitment to the translocon in Saccharomyces cerevisiae.
We studied the molecular nature of the interaction between the integral membrane protein Sec63p and the lumenal Hsp70 BiP to elucidate their role in the process of precursor transit into the ER of Saccharomyces cerevisiae. A lumenal stretch of Sec63p with homology to the Escherichia coli protein DnaJ is the likely region of interface between Sec63p and BiP. This domain, purified as a fusion protein (63Jp) with glutathione S-transferase (GST), mediated a stable ATP-dependent binding interaction between 63Jp and BiP and stimulated the ATPase activity of BiP. The interaction was highly selective because only BiP was retained on immobilized 63Jp when detergent-solubilized microsomes were mixed with ATP and the fusion protein. GST alone was inactive in these assays. Additionally, a GST fusion containing a point mutation in the lumenal domain of Sec63p did not interact with BiP. Finally, we found that the soluble Sec63p lumenal domain inhibited efficient precursor import into proteoliposomes reconstituted so as to incorporate both BiP and the fusion protein. We conclude that the lumenal domain of Sec63p is sufficient to mediate enzymatic interaction with BiP and that this interaction positioned at the translocation apparatus or translocon at the lumenal face of the ER is vital for protein translocation into the ER.


A simple correction for B1 field errors in magnetization transfer ratio measurements.
B1 errors are a problem in magnetization transfer ratio (MTR) measurements because the MTR value is dependent on the amplitude of the magnetization transfer (MT) pulse. B1 errors can arise from radiofrequency (RF) nonuniformity (caused by the RF coil, or skin effect and dielectric resonance in the subject's head) and also from inaccurate setting of the transmitter output when compensating for varying amounts of loading of the RF coil. B1 errors, and hence MTR errors, may be up to 5-10%, a large source of error in quantitative MR measurements. Radiofrequency nonuniformity may cause MTR histograms to be broadened. The dependence of MTR on B1 was modeled using binary spin bath theory, with a continuous wave (CW) approximation. For B1 reductions of up to 20%, normalized plots for different brain tissue types could be approximated by a single line, indicating that a systematic correction could be applied to MTR measurements with a known B1 error, regardless of tissue type. On a 1.5-T scanner with a birdcage coil, MTR was measured in 18 tissue types in five controls. The MT pulse amplitude was reduced in steps from its nominal value by up to 20%. Averaging data over all controls and tissue types resulted in a line fitting mtr(normalized)=0.812b(1normalized)+0.193, where mtr(normalized) is the normalized value of MTR (relative to its value at the nominal B1) and b(1normalized) is the normalized value of B1 (relative to its nominal value). For a 20% reduction in MT pulse amplitude (i.e., b(1normalized)=0.80), the mean MTR value for the 18 tissue types was 7.0 percent units (pu) below the correct value. After correction using the single equation above for all tissue types, all MTR values were within 1.5 pu of their correct value [root mean square (rms) error=0.7 pu]. Magnetization transfer ratio values tended to be slightly overcorrected because the simple linear correction scheme is only an approximation to the true MTR dependence on B1. A B1 field mapping technique was implemented, based on the double angle method (DAM), with fast spin-echo (FSE) readout, and TR=15 s; this took a total of 6 min of imaging time. This was used to quantify B(1) errors and correct MTR maps and histograms. However, the cerebrospinal fluid (CSF) T1 is very long (approximately 4.2 s); thus, to achieve complete longitudinal relaxation (a requirement of the DAM B1 mapping method), an increase in TR and, hence, acquisition time would be required. In general, however, we are not interested in calculating the B1 in the CSF, although it is important that the B1 is determined in partial volume voxels around the CSF. Using our birdcage head coil, whole-brain B1 histograms were found to have full-width half maximums (FWHMs) ranging from just 6.8% to 11.5% of the nominal B1 value. The FSE DAM B1 field mapping technique was shown to be robust, although a longer TR time may be desirable to ensure complete elimination of CSF partial volume errors. The procedure can be applied on any scanner where the Euro-MT sequence is available, or alternatively, where the amplitude of B1 or of the MT pulse can be manually reduced in order to perform this type of "calibration" experiment for the particular MTR sequence used. The MTR is known to be highly dependent on the parameters of the sequence used, in particular, the MT pulse shape, flip angle, duration, and offset frequency, and the repetition time TR' between successive MT pulses. Therefore, correction schemes will differ for different MTR sequences, and new data sets would be required to calculate these different correction schemes.


The redox antimalarial dihydroartemisinin targets human metastatic melanoma cells but not primary melanocytes with induction of NOXA-dependent apoptosis.
Recent research suggests that altered redox control of melanoma cell survival, proliferation, and invasiveness represents a chemical vulnerability that can be targeted by pharmacological modulation of cellular oxidative stress. The endoperoxide artemisinin and semisynthetic artemisinin-derivatives including dihydroartemisinin (DHA) constitute a major class of antimalarials that kill plasmodium parasites through induction of iron-dependent oxidative stress. Here, we demonstrate that DHA may serve as a redox chemotherapeutic that selectively induces melanoma cell apoptosis without compromising viability of primary human melanocytes. Cultured human metastatic melanoma cells (A375, G361, LOX) were sensitive to DHA-induced apoptosis with upregulation of cellular oxidative stress, phosphatidylserine externalization, and activational cleavage of procaspase 3. Expression array analysis revealed DHA-induced upregulation of oxidative and genotoxic stress response genes (GADD45A, GADD153, CDKN1A, PMAIP1, HMOX1, EGR1) in A375 cells. DHA exposure caused early upregulation of the BH3-only protein NOXA, a proapototic member of the Bcl2 family encoded by PMAIP1, and genetic antagonism (siRNA targeting PMAIP1) rescued melanoma cells from apoptosis indicating a causative role of NOXA-upregulation in DHA-induced melanoma cell death. Comet analysis revealed early DHA-induction of genotoxic stress accompanied by p53 activational phosphorylation (Ser 15). In primary human epidermal melanocytes, viability was not compromised by DHA, and oxidative stress, comet tail moment, and PMAIP1 (NOXA) expression remained unaltered. Taken together, these data demonstrate that metastatic melanoma cells display a specific vulnerability to DHA-induced NOXA-dependent apoptosis and suggest feasibility of future anti-melanoma intervention using artemisinin-derived clinical redox antimalarials.


Epiretinal Membrane Surgery Outcomes in Eyes with Subretinal Drusenoid Deposits: A Case Control Study.
To evaluate outcomes of epiretinal membrane (ERM) surgery in eyes with subretinal drusenoid deposits (SRDDs) and to compare them with those with isolated ERM. Retrospective case-control study of consecutive patients who underwent pars plana vitrectomy (PPV) with ERM peeling. Twenty-five patients with SRDDs on spectral-domain (SD)-OCT who underwent surgery for ERM were included in the study. From the same cohort, for each case, we selected 2 age-matched control participants (50 eyes with isolated ERM). Preoperative best-corrected visual acuity (BCVA) also was matched as closely as possible. All participants underwent PPV and ERM peel for primary ERM. Postoperative BCVA, improvement in BCVA, preoperative and postoperative central macular thickness, surgical complications, and development of age-related macular degeneration (AMD) were recorded. At final examination, mean postoperative BCVA was significantly less for eyes with SRDDs (0.51 logarithm of the minimal angle of resolution [logMAR] vs. 0.21 logMAR; P = 0.0001). Eyes with SRDDs demonstrated less improvement in BCVA after ERM surgery (0.13 logMAR vs. 0.30 logMAR; P = 0.0032). Eyes with SRDDs were significantly less likely to gain 2 or more Snellen lines of BCVA after ERM surgery (28% vs. 56%; P = 0.028). Three of 25 patients (12%) undergoing ERM surgery showed worsening of Snellen BCVA by 2 lines or more. Three of 25 eyes (12%) with SRDDs demonstrated advanced AMD after surgery, compared with 0 participants in the control group (P = 0.034). Epiretinal membrane surgery in eyes with SRDDs is associated with less favorable visual outcomes. Fewer patients demonstrate gain in BCVA, whereas a significant number show a deleterious decline. After surgery, AMD incidence seems high and patients may have an increased risk of raised intraocular pressure. These findings require further study to establish whether this represents a causal relationship. Surgeons should be vigilant for these complications. Appropriate patient counseling during the consenting process must be made.


Positive airway pressure levels and pneumothorax: a case-control study in very low birth weight infants.
Evaluate the incidence of pneumothorax (PTx) and the levels of positive airway pressure (Paw) applied to very low birth weight infants during the first 5 days of life (DOL), after evidence-based protocols using early continuous positive airway pressure (CPAP) and high levels of Paw (CPAP or mean airway pressure) were implemented. From 2004 to 2007, all infants submitted to assisted ventilation that developed PTx were identified. Controls were matched by birth weight, gestational age, and type of ventilatory support. Paw levels were averaged on a time-weighted basis. A p value <0.05 was considered significant. A total of 25 infants developed PTx (3.8%); 23 during the first 5 DOL. PTx was diagnosed at 14 h of life (1.3-80 h) when 74% were treated with mechanical ventilation. In controls, Paw decreased over time whereas in PTx infants it did not decline until after 80 h. PTx infants had an increase in Paw from 12 h up to 6 h prior to the diagnosis. The rate of PTx was low even after the implementation of the protocols. An association between Paw levels and PTx was observed but until the precise time of onset of a PTx can be determined this should be regarded either as an early signal or as an indicator of more severe lung disease.


BMP4 administration induces differentiation of CD133+ hepatic cancer stem cells, blocking their contributions to hepatocellular carcinoma.
CD133+ cancer stem cells (CSC) contribute to hepatocellular carcinoma (HCC) progression and resistance to therapy. Bone morphogenetic protein BMP4 plays an important role in hepatogenesis and hepatic stem cell differentiation, but little is known about its function in hepatic CSCs. In this study, we showed that high-dose exogenous BMP4 promotes CD133+ HCC CSC differentiation and inhibits the self-renewal, chemotherapeutic resistance, and tumorigenic capacity of these cells. Interestingly, we found that low-dose exogenous BMP4 upregulated CD133 protein expression in vitro, and endogenous BMP4 was preferentially expressed in CD133+ HCC CSCs, suggesting that low doses of BMP4 may facilitate CSC maintenance. A reduction in endogenous BMP4 levels decreased CD133 protein expression in vitro. In HCC tissues, expression of the BMP4 signaling target gene SMAD6 was positively correlated with CD133 expression. Activation of the Erk1/2 signaling pathway led to BMP4-mediated reduction in CD133 expression, which was reversed by treatment with MEK inhibitors. Taken together, our findings indicated that BMP4 might be a potent therapeutic agent in HCC that targets CSCs.


Epidemiology of sexually transmitted diseases among pregnant adolescents.
The purpose of this study was to determine the epidemiology of sexually transmitted diseases (STDs) among pregnant adolescents. Charts of all patients (n = 735) who attended the Maternal and Infant Care Clinic at University Hospital, Newark, NJ, between July 1, 1991, and June 30, 1992, were reviewed for STDs which included gonorrhea, chlamydia, syphilis, and human immunodeficiency virus (HIV). At the first prenatal visit, each registrant had endocervical specimens obtained to detect gonorrhea and chlamydia. A serum sample was obtained for syphilis screening. HIV testing was made available to all patients and testing was done on a voluntary basis. The same STD screening that was done at the initial visit was repeated at 28 and 36 weeks. Twenty-five percent of patients tested positive for one or more STDs. The mean patient age was 17.3 years. The mean gestational age at first visit was 19.5 weeks. The mean number of visits was 7.3. The following STDs were identified: 4.8% of patients tested positive for gonorrhea, 20.9% tested positive for chlamydia, and 1.7% tested positive for syphilis. Twenty-one percent of patients had a positive STD diagnosed at the initial visit. Another 4.8% of patients had an STD diagnosed at some time after the initial visit when the initial screen was negative for STDs. An additional 1% of patients who initially tested positive for an STD had subsequent screening which revealed another STD (different organism). Seven patients tested HIV positive. Sixty-one percent of patients with STDs agreed to HIV testing. One patient had HIV coexistent with another STD. Pregnant adolescents are at risk for multiple STDs. HIV testing should be offered. STD screening should be repeated in the third trimester in adolescent patients.


The incidence of induced abortion in Malawi.
Abortion is legally restricted in Malawi, and no data are available on the incidence of the procedure. The Abortion Incidence Complications Methodology was used to estimate levels of induced abortion in Malawi in 2009. Data on provision of postabortion care were collected from 166 public, nongovernmental and private health facilities, and estimates of the likelihood that women who have abortions experience complications and seek care were obtained from 56 key informants. Data from these surveys and from the 2010 Malawi Demographic and Health Survey were used to calculate abortion rates and ratios, and rates of pregnancy and unintended pregnancy. Approximately 18,700 women in Malawi were treated in health facilities for complications of induced abortion in 2009. An estimated 67,300 induced abortions were performed, equivalent to a rate of 23 abortions per 1,000 women aged 15-44 and an abortion ratio of 12 per 100 live births. The abortion rate was higher in the North (35 per 1,000) than in the Central region or the South (20-23 per 1,000). The unintended pregnancy rate in 2010 was 139 per 1,000 women aged 15-44, and an estimated 52% of all pregnancies were unintended. Unsafe abortion is common in Malawi. Interventions are needed to help women and couples avoid unwanted pregnancy, reduce the need for unsafe abortion and decrease maternal mortality.


Reproductive isolation during domestication.
It has been hypothesized that reproductive isolation should facilitate evolution under domestication. However, a systematic comparison of reproductive barrier strength between crops and their progenitors has not been conducted to test this hypothesis. Here, we present a systematic survey of reproductive barriers between 32 economically important crop species and their progenitors to better understand the role of reproductive isolation during the domestication process. We took a conservative approach, avoiding those types of reproductive isolation that are poorly known for these taxa (e.g., differences in flowering time). We show that the majority of crops surveyed are isolated from their progenitors by one or more reproductive barriers, despite the fact that the most important reproductive barrier in natural systems, geographical isolation, was absent, at least in the initial stages of domestication for most species. Thus, barriers to reproduction between crops and wild relatives are closely associated with domestication and may facilitate it, thereby raising the question whether reproductive isolation could be viewed as a long-overlooked "domestication trait." Some of the reproductive barriers observed (e.g., polyploidy and uniparental reproduction), however, may have been favored for reasons other than, or in addition to, their effects on gene flow.


Cytomegalovirus Virions Shed in Urine Have a Reversible Block to Epithelial Cell Entry and Are Highly Resistant to Antibody Neutralization.
Cytomegalovirus (CMV) causes sensorineural hearing loss and developmental disabilities in newborns when infections are acquired in utero Pregnant women may acquire CMV from oral exposure to CMV in urine or saliva from young children. Neutralizing antibodies in maternal saliva have the potential to prevent maternal infection and, in turn, fetal infection. As CMV uses different viral glycoprotein complexes to enter different cell types, the first cells to be infected in the oral cavity could determine the type of antibodies needed to disrupt oral transmission. Antibodies targeting the pentameric complex (PC) should block CMV entry into epithelial cells but not into fibroblasts or Langerhans cells (which do not require the PC for entry), while antibodies targeting glycoprotein complexes gB or gH/gL would be needed to block entry into fibroblasts, Langerhans cells, or other cell types. To assess the potential for antibodies to disrupt oral acquisition, CMV from culture-positive urine samples (uCMV) was used to study cell tropisms and sensitivity to antibody neutralization. uCMV entered epithelial cells poorly compared with the entry into fibroblasts. CMV-hyperimmune globulin or monoclonal antibodies targeting gB, gH/gL, or the PC were incapable of blocking the entry of uCMV into either fibroblasts or epithelial cells. Both phenotypes were lost after one passage in cultured fibroblasts, suggestive of a nongenetic mechanism. These results suggest that uCMV virions have a reversible block to epithelial cell entry. Antibodies may be ineffective in preventing maternal oral CMV acquisition but may limit viral spread in blood or tissues, thereby reducing or preventing fetal infection and disease.


A diabetes registrar assisted workflow intervention in general practice for systematic initiation of cardiorenal medications for patients with type 2 diabetes and albuminuria in Aotearoa New Zealand.
To evaluate whether a weekly diabetes registrar clinic and case discussions conducted over 12 weeks in primary care improves guideline management of type 2 diabetes (T2D). A registrar-led diabetes clinic was incorporated into two primary care practices in Tāmaki Makaurau Auckland for 3 months. Patients with T2D and albuminuria appearing on practice dashboards as not prescribed angiotensin-converting enzyme inhibitor/angiotensin receptor blocker (ACEi/ARB), or sodium-glucose cotransporter-2 inhibitor/glucagon-like peptide-1 receptor agonist (SGLT2i/GLP1RA) were booked into these clinics. Opportunistic education sessions were provided by the diabetes registrar and prescribers were surveyed to understand the challenges in management of T2D. Of 125 patients booked, 80 attended the registrar clinic. Of these, 68% were clinically suitable for SGLT2i/GLP1RA and 8% for ACEi/ARB. SGLT2i/GLP1RA were initiated in 92% and ACEi/ARB in 89% of eligible patients. Two patients had contraindications for SGLT2i/GLP1RA, and one patient declined both. Additional cardiorenal medications were initiated in 16% of patients. Survey responses of 12 prescribers indicated acute illness takes priority over diabetes management, and lack of time and knowledge are main barriers to optimising diabetes care. A visiting diabetes registrar intervention was successful in initiating guideline medications for T2D in primary care. It remains to be evaluated whether this leads to practice-wide improvements in prescribing gaps in the short or longer term.


Association between depression and healthcare expenditures among elderly cancer patients.
Both depression and cancer are economically burdensome. However, how depression affects the healthcare expenditures of elderly cancer patients from payers' and patients' perspectives is largely unknown. This study investigated whether depression resulted in higher healthcare expenditures among these patients from both payers' and patients' perspectives and identified health service use categories associated with increased expenditures. From the Medicare Current Beneficiary Survey (MCBS)-Medicare database, we identified breast, lung and prostate cancer patients aged 65 years and over who were newly diagnosed between 2007 and 2012. Presence of depression was based on self-reports from the surveys. We used generalized linear models (GLM) and two-part models to examine the impact of depression on healthcare expenditures during the first two years of cancer diagnosis controlling for a vast array of covariates. We stratified the analyses of total healthcare expenditures by healthcare services and payers. Out of the 710 elderly breast, lung and prostate cancer patients in our study cohort, 128 (17.7%) reported depression. Individuals with depression had $11,454 higher total healthcare expenditures, $8213 higher medical provider expenditures and $405 higher other services expenditures compared to their counterparts without depression. Also, they were significantly more likely to have inpatient services. For payers, they incurred $8280 and $1270 higher expenditures from Medicare's and patients' perspectives, respectively. Elderly cancer patients with depression have significantly higher healthcare expenditures from both payers' and patients' perspectives and over different expenditure types. More research is needed in depression screening, diagnosis and treatment for this population.


Neuroinflammation impact in epileptogenesis and new treatment strategy.
Epilepsy is considered a major serious chronic neurological disorder, characterized by recurrent seizures. It is usually associated with a history of a lesion in the nervous system. Irregular activation of inflammatory molecules in the injured tissue is an important factor in the development of epilepsy. It is unclear how the imbalanced regulation of inflammatory mediators contributes to epilepsy. A recent research goal is to identify interconnected inflammation pathways which may be involved in the development of epilepsy. The clinical use of available antiepileptic drugs is often restricted by their limitations, incidence of several side effects, and drug interactions. So development of new drugs, which modulate epilepsy through novel mechanisms, is necessary. Alternative therapies and diet have recently reported positive treatment outcomes in epilepsy. Vitamin D (Vit D) has shown prophylactic and therapeutic potential in different neurological disorders. So, the aim of current study was to review the associations between different brain inflammatory mediators and epileptogenesis, to strengthen the idea that targeting inflammatory pathway may be an effective therapeutic strategy to prevent or treat epilepsy. In addition, neuroprotective effects and mechanisms of Vit D in clinical and preclinical studies of epilepsy were reviewed.