TERT promoter mutations and gene amplification: promoting TERT expression in Merkel cell carcinoma.
Telomerase activation through the induction of its catalytic component TERT is essential in carcinogenesis. The regulatory mechanism and clinical significance underlying cancer-specific TERT expression have been extensively investigated in various human malignancies, but little is known about these in Merkel cell carcinoma (MCC), an aggressive neuroendocrine skin tumor. Here we addressed these issues by determining TERT promoter mutations, gene amplification, mRNA expression and association with clinical variables in MCC. TERT mRNA was expressed in 6/6 MCC cell lines and 41 of 43 tumors derived from 35 MCC patients. Telomerase activity was detectable in all 6 cell lines and 11 tumors analyzed. TERT promoter mutations were identified in 1/6 cell lines and 4/35 (11.4%) MCC cases. The mutation exhibited UV signature and occurred in sun-exposed areas. Increased TERT gene copy numbers were observed in 1/6 cell lines and 11/14 (79%) tumors, and highly correlated with its mRNA expression (r = 0.7419, P = 0.0024). Shorter overall survival was significantly associated with higher TERT mRNA levels in MCC patients (P = 0.032). Collectively, TERT expression and telomerase activity is widespread in MCC, and may be attributable to TERT promoter mutations and gene amplification. Higher TERT expression predicts poor patient outcomes.


Long-term sick leave and its risk factors during pregnancy among Danish hospital employees.
The authors sought to describe risk indicators of long-term sick leave during pregnancy among hospital employees. A register-based study was undertaken of 4,852 female hospital employees aged 20-45 years from the second largest hospital in Denmark during 1995-99 based on job titles, working time, sick leave, and births combined with a survey among a total of 773 women who had been pregnant during their employment (response rate 85%). Altogether 236 (31%) were on sick leave for at least 10% of their scheduled work time during their latest pregnancy and 169 (22%) had been absent at least 20% of the time. The pregnant women had an average sickness absence of 6.1 days per month, non-pregnant women 0.95 days per month. Sick leave was more frequent in late than in early gestation. Women employed as nursing aides or hospital orderlies, launderers, and nurses had more sick leave days than other hospital employees. Part-time work, previous sickness absence not related to pregnancy, and previous chronic back pain were risk factors for long-term sick leave as were much walking or standing, long working days, high work level, little practical support from supervisors and colleagues, low job control, much lifting and night or shift work. Sick leave was unrelated to family size, support from the family and number of working years. Long-term sick leave during pregnancy was frequent and to some extent predictable. Efforts should be made to organize work for pregnant women in a manner that optimizes their health and well-being.


Indole-2-carboxamidines as novel NR2B selective NMDA receptor antagonists.
A novel series of indole-2-carboxamidine derivatives was prepared and identified as NR2B selective NMDA receptor antagonists. The influence of the substituents on the indole skeleton as well as the substitution of the benzyl moiety on the biological activity of the compounds was studied. Compound 5a was po active in the formalin test in mouse.


Characteristics and incidence of large eggs in Trichuris muris.
The production of small numbers of large eggs among the standard-sized eggs of Trichuris trichiura is well known. Large eggs have also been observed in Trichuris muris, but they have not been studied previously. This paper compares the characteristics of the large eggs (LEs, ≥74.5 μm long) and standard-sized eggs (SEs, <74.5 μm long) in cultures of T. muris. Among 112,554 cultured eggs, LEs occurred at very low frequency (0.03 %, i.e., about three large eggs per 10(4) cultured eggs). Embryonated eggs represented 93.72 % of SEs, but only 25.00 % of LEs were embryonated. Embryonated LEs and SEs contained fully matured larvae. An atypical category of unembryonated egg, which contained an incompletely developed larva, an abnormal larva, or granular components, was common among the LEs. However, similar atypical unembryonated SEs were rarely observed. These observations suggest that the LEs that occur very infrequently in T. muris result from an abnormality of embryonation (larval development).


RNA interference screening identifies a novel role for PCTK1/CDK16 in medulloblastoma with c-Myc amplification.
Medulloblastoma (MB) is the most common malignant brain tumor in children and is associated with a poor outcome. cMYC amplification characterizes a subgroup of MB with very poor prognosis. However, there exist so far no targeted therapies for the subgroup of MB with cMYC amplification. Here we used kinome-wide RNA interference screening to identify novel kinases that may be targeted to inhibit the proliferation of c-Myc-overexpressing MB. The RNAi screen identified a set of 5 genes that could be targeted to selectively impair the proliferation of c-Myc-overexpressing MB cell lines: AKAP12 (A-kinase anchor protein), CSNK1α1 (casein kinase 1, alpha 1), EPHA7 (EPH receptor A7) and PCTK1 (PCTAIRE protein kinase 1). When using RNAi and a pharmacological inhibitor selective for PCTK1, we could show that this kinase plays a crucial role in the proliferation of MB cell lines and the activation of the mammalian target of rapamycin (mTOR) pathway. In addition, pharmacological PCTK1 inhibition reduced the expression levels of c-Myc. Finally, targeting PCTK1 selectively impaired the tumor growth of c-Myc-overexpressing MB cells in vivo. Together our data uncover a novel and crucial role for PCTK1 in the proliferation and survival of MB characterized by cMYC amplification.


Use of the esophageal lead in the diagnosis of mechanisms of reciprocating supraventricular tachycardia.
Recent studies have emphasized the role of concealed accessory pathways in reciprocating supraventricular tachycardia. Diagnosis has generally required multicatheter electrophysiologic study. We recorded esophageal electrograms during study in 16 patients with reciprocating tachycardia due to reentry using an accessory atrioventricular pathway, and in 12 patients with reciprocating tachycardia due to reentry in the AV node. The interval from onset of ventricular depolarization to earliest atrial activation (V-AMIN), earliest atrial activity on the esophageal lead (V-AESO), and high right atrium (V-HRA) was measured. No patient with RT due to an accessory atrioventricular pathway had a V-AMIN or V-AESO less than 70 ms, or a V-HRA less than 95 ms. In contrast, 11 of 12 patients with reentry in the AV node had V-AESO intervals less than 70 ms. Esophageal recording during reciprocating tachycardia provides a simple screening procedure available to all practicing physicians to exclude the diagnosis of accessory atrioventricular pathways in the genesis of paroxysmal supraventricular tachycardia.


Efficient Nanostructuring of Silicon by Electrochemical Alloying/Dealloying in Molten Salts for Improved Lithium Storage.
Application of nanostructured silicon (nSi) is significantly retarded by challenges in the production of affordable nSi. We herein report a high-yield (ca. 100 %) and low-energy (2 kWh Kg-nSi-1 ) nanostructuring of industrial microsized silicon (mSi) through a closed-loop electrochemical Mg alloying/dealloying in molten MgCl2 /NaCl/KCl at 773 K. The resulting nSi unexpectedly shows a salt-unwetted character, allowing an automatic separation from the melts. Thus water washing and accompanying oxidation of the nSi can be avoided. The final product has a nanoporous structure and comprises Si nanorods (ca. 30 nm in diameter) with an ultrathin oxide coating. It can be used for Li storage giving a combination of high initial coulombic efficiency, high specific capacity, and long cycling stability. This nanostructuring process consumes very few chemicals except for the mSi and produces almost zero waste.


Statistical inference of the relative concentration index for complex surveys.
The relative concentration index is a widely used measure for assessing relative differences in health across all socioeconomic population groups. We extend its usage to individual-level data collected through complex surveys by deriving its variance using the Taylor linearization (TL) method. Two existing plug-in variance estimators that only require grouped data are also compared. We discuss sources of uncertainty that each variance estimator considers and present simulation studies to compare the performance of the three estimators under various sampling designs. The proposed TL variance estimator consistently produces valid results; however, it requires the access to individual-level data. Both plug-in variance estimators are biased because of failure to account for certain error sources. However, when only grouped data is available, one of the plug-in estimators can be valid as long as the socioeconomic groups are treated equally sized, a commonly used analytic strategy to emphasize group's instead of individual's burden of disease in health disparity assessment. We illustrate the three variance estimators by applying them to assessing socioeconomic disparities in child and adolescent obesity using complex survey sampled drawn from the National Health and Nutrition Examination Survey.


Estimation of disruption of animal cells by laminar shear stress.
Using mechanical cell properties measured by micromanipulation, and a model of cell distortion in laminar flow fields, a method has been developed for predicting disruption of animal cells by laminar shear stresses. Predictions of the model were compared with measured losses of cell number and viability of TB/C3 murine hybridomas sheared in a cone and plate viscometer at shear rates up to 3950 s(-1), and shear stresses up to 600 Nm(-2), achieved by enhancement of viscosity with dextran. In all cases, the experimental, results and predictions were within 30%. Such excellent agreement suggests it might be possible to use micromanipulation measurements of animal cell mechanical properties to predict cell damage in more complex flow fields, such as those in bioreactors.


Removal of Cd(II), Cu(II) and Zn(II) from aqueous solutions by live Phanerochaete chrysosporium.
Living Phanerochaete chrysosporium mycelia were used to remove heavy metals of Cd(II), Cu(II) and Zn(II) in auqeous solution. The uptake of heavy metal by the mycelia was dependent on the environmental conditions. The optimum biosorption conditions of Cd(II), Cu(II), and Zn(II) were pH 5.5-6.5 at 37 degrees C, and 6 h. Under these conditions, the fungal biosorbent removed Cd(II), Cu(II), and Zn(II) rapidly and efficiently with maximum metal removal capacities of 59.77 mg/g, 74.78 mg/g, and 54.12 mg/g, respectively. The pseudo second-order kinetic model was superior to the pseudo first-order kinetic model, which indicated that the change ofthe surface sorption sites number was proportional to the square ofthe remaining unoccupied surface sites number. Fourier transform infrared (FT-IR) spectra indicated that hydroxyl and carboxyl groups were relevant to biosorption. Some crystal particles were found on the surface of the P. chrysosporium under scanning electron microscopy. Energy dispersive X-ray analysis and FT-IR revealed that amino acids and proteins were involved in binding metal ions. The results demonstrated that P. chrysosporium was a good potential biosorbent for adsorbing heavy metals.


Acquisition of immunity in mothers of infants administered trivalent oral poliovirus vaccine.
The hypothesis that live oral poliovirus vaccine (OPV) confers immunity to persons in close contact with the vaccine recipient was tested by comparing cord blood antibody titers in 31 first-born neonates to those of 48 neonates with one or more siblings adequately immunized with OPV. A borderline or negative result (defined as a reciprocal titer of < or = 8) for at least one type was significantly more prevalent among first-born neonates than among neonates with one or more siblings [17/31 (55%) versus 17/48 (27%) respectively; p < 0.03]. This difference was consistent for all three poliovirus types. The geometric mean titer (GMT) was consistently higher for each serotype in infants with one or more siblings compared with first-born neonates: 134.9 versus 64.5 for poliovirus 1; 262.1 versus 95.6 for poliovirus 2; and 48.6 versus 19.4 for poliovirus 3, respectively. When cord blood of neonates with two or more siblings was compared to that of neonates with only one sibling, no difference in titers was observed. Since mothers of one or more infants were on average older and less educated, the results were adjusted accordingly, but the same trend was observed again. These findings support the notion that OPV is important, not only as a vaccine for the individual infants, but also as a means of conferring immunity to persons in close contact.


[Lytic enzymes of the pneumococcal system].
The cell wall lytic enzymes of Streptococcus pneumoniae and its bacteriophages provide a reliable experimental model to support the extended idea concerning the modular organization of most proteins. The comparative analysis of the deduced amino acid sequences of these enzymes, as well as the construction of functional chimeric phage-bacterial enzymes, demonstrate that the C-terminal domain, which contains a large number of repeated amino acid motifs, is the substrate-binding domain, whereas the N-terminal domain provides enzymatic specificity. The development of chimeric proteins from genes encoding enzymes that specifically degrade cell wall of bacteria belonging to different genera (e.g. Streptococcus and Clostridium), supports the hypothesis concerning domainal interchange as a driving force for the construction of novel functional proteins that bacteria might create to allow them to become adapted to new environmental situations.


The difference in pharmacokinetics and pharmacodynamics between extended-release fluvastatin and immediate-release fluvastatin in healthy Chinese subjects.
The aim of this study was to evaluate the difference in pharmacokinetics and pharmacodynamics between extended-release (ER) fluvastatin tablet and its immediate-release (IR) capsule in Chinese healthy subjects. This was an open-label, single/multiple-dose, two-period, two-treatment, crossover, randomized trial with a minimum washout period of 7 days. Twenty healthy male adult subjects were given fluvastatin ER tablet 80 mg QD by oral administration or fluvastatin IR capsule 40 mg BID for seven days. Blood samples were collected up to 24 hours after dosing on day 1 and day 7. Serum concentrations of fluvastatin were determined by LC-MS/MS. For fluvastatin ER tablet 80 mg QD, C(max) was 61.0 ± 39.0 and 63.9 ± 29.7 ng/mL, and AUC(0-24 h) was 242 ± 156 and 253 ± 91.1 ng·h/mL on day 1 and 7, respectively. For fluvastatin IR capsule 40 mg BID, C(max) was 283 ± 271 and 382 ± 255 ng/mL, and AUC(0-24 h) was 720 ± 776 and 917 ± 994 ng·h/mL on day 1 and day 7, respectively. The relative bioavailability of fluvastatin ER tablet 80 mg QD to fluvastatin IR capsule 40 mg BID is (45.3 ± 23.9)% and (43.3 ± 24.1)% on day 1 and day 7, respectively. T(max) for fluvastatin ER tablet was 2.50 and 2.60 h and for capsule was 0.78 and 0.88 h on day 1 and day 7, respectively. In the first period, compared to baseline, cholesterol decreased 15.3% in fluvastatin ER tablet 80 mg QD and 16.9% in fluvastatin IR capsule 40 mg BID. Triglyceride decreased 3.7% in fluvastatin ER tablet 80 mg QD and 19.1% in fluvastatin IR capsule 40 mg BID. The difference has no statistical significance at P > 0.05 in reduction percent of cholesterol and triglyceride between the two groups. No adverse events were recorded. The results indicated that C(max) of fluvastatin ER tablet is reduced and T(max) is prolonged compared with IR capsule. There is no accumulation for ER formulation after multiple doses.


Analysis of experts' quantitative assessment of adolescent basketball players and the role of anthropometric and physiological attributes.
(1) can measurements of anthropometric and physiological attributes substitute for expert assessment of adolescent basketball players, and (2) how much does the quantitative assessment of a player vary among experts? The first question is relevant to the potential simplification of the player selection process. The second question pertains directly to the validity of expert quantitative assessment. Our research was based on data from 148 U14 female and male basketball players. For each player, an array of anthropometric and physiological attributes was recorded, including body height, body mass, BMI, and several motor skill tests. Furthermore, each player's current ability and potential ability were quantitatively evaluated by two different experts from a group of seven experts. Analysis of the recorded data showed that the anthropometric and physiological attributes explained between 15% and 40% of the variance in experts' scores. The primary predictive attributes were speed and agility (for predicting current ability) and body height and growth potential (for predicting potential ability). We concluded that these attributes were not sufficiently informative to act as a substitute for expert assessment of the players' current or potential ability. There is substantial variability in different experts' scores of the same player's ability. However, the differences between experts are mostly in scale, and the relationships between experts' scores are monotonic. That is, different experts rank players on ability very similarly, but their scores are not well calibrated.


XBP1 signalling is essential for alleviating mutant protein aggregation in ER-stress related skeletal disease.
The unfolded protein response (UPR) is a conserved cellular response to the accumulation of proteinaceous material in endoplasmic reticulum (ER), active both in health and disease to alleviate cellular stress and improve protein folding. Multiple epiphyseal dysplasia (EDM5) is a genetic skeletal condition and a classic example of an intracellular protein aggregation disease, whereby mutant matrilin-3 forms large insoluble aggregates in the ER lumen, resulting in a specific 'disease signature' of increased expression of chaperones and foldases, and alternative splicing of the UPR effector XBP1. Matrilin-3 is expressed exclusively by chondrocytes thereby making EDM5 a perfect model system to study the role of protein aggregation in disease. In order to dissect the role of XBP1 signalling in aggregation-related conditions we crossed a p.V194D Matn3 knock-in mouse model of EDM5 with a mouse line carrying a cartilage specific deletion of XBP1 and analysed the resulting phenotype. Interestingly, the growth of mice carrying the Matn3 p.V194D mutation compounded with the cartilage specific deletion of XBP1 was severely retarded. Further phenotyping revealed increased intracellular retention of amyloid-like aggregates of mutant matrilin-3 coupled with dramatically decreased cell proliferation and increased apoptosis, suggesting a role of XBP1 signalling in protein accumulation and/or degradation. Transcriptomic analysis of chondrocytes extracted from wild type, EDM5, Xbp1-null and compound mutant lines revealed that the alternative splicing of Xbp1 is crucial in modulating levels of protein aggregation. Moreover, through detailed transcriptomic comparison with a model of metaphyseal chondrodysplasia type Schmid (MCDS), an UPR-related skeletal condition in which XBP1 was removed without overt consequences, we show for the first time that the differentiation-state of cells within the cartilage growth plate influences the UPR resulting from retention of a misfolded mutant protein and postulate that modulation of XBP1 signalling pathway presents a therapeutic target for aggregation related conditions in cells undergoing proliferation.


The sites of cleavage on Escherichia coli glutamine synthetase by dithiothreitol, Fe(III) and O2.
Native conformation of an enzyme molecule is required for the specific non-enzymatic cleavage of Escherichia coli glutamine synthetase by a metal-catalyzing oxidation system comprised of dithiothreitol, Fe(III) and O2. The cleavage reaction is greatly inhibited by the addition of Mg(II). Two major cleavage sites are identified between amino acid residues 264 and 268, and roughly between amino acid residues 31 and 34, which are located on the protein segments forming the active site of the enzyme. These results suggest that the cleavage reaction is a largely site-specific process involving active oxygen species generated at the divalent cation binding sites on glutamine synthetase.


Loss of Function of the E1-Like-b Gene Associates With Early Flowering Under Long-Day Conditions in Soybean.
Photoperiod response of flowering determines plant adaptation to different latitudes. Soybean, a short-day plant, has gained the ability to flower under long-day conditions during the growing season at higher latitudes, mainly through dysfunction of phytochrome A genes (E3 and E4) and the floral repressor E1. In this study, we identified a novel molecular genetic basis of photoperiod insensitivity in Far-Eastern Russian soybean cultivars. By testcrossing these cultivars with a Canadian cultivar Harosoy near-isogenic line for a recessive e3 allele, followed by association tests and fine mapping, we determined that the insensitivity was inherited as a single recessive gene located in an 842-kb interval in the pericentromeric region of chromosome 4, where E1-Like b (E1Lb), a homoeolog of E1, is located. Sequencing analysis detected a single-nucleotide deletion in the coding sequence of the gene in insensitive cultivars, which generated a premature stop codon. Near-isogenic lines (NILs) for the loss-of-function allele (designated e1lb) exhibited upregulated expression of soybean FLOWERING LOCUS T (FT) orthologs, FT2a and FT5a, and flowered earlier than those for E1Lb under long-day conditions in both the e3/E4 and E3/E4 genetic backgrounds. These NILs further lacked the inhibitory effect on flowering by far-red light-enriched long-day conditions, which is mediated by E4, but not that of red-light-enriched long-day conditions, which is mediated by E3. These findings suggest that E1Lb retards flowering under long-day conditions by repressing the expression of FT2a and FT5a independently of E1. This loss-of-function allele can be used as a new resource in breeding of photoperiod-insensitive cultivars, and may improve our understanding of the function of the E1 family genes in the photoperiod responses of flowering in soybean.


Structure-guided design enables development of a hyperpolarized molecular probe for the detection of aminopeptidase N activity in vivo.
Dynamic nuclear polarization (DNP) is a cutting-edge technique that markedly enhances the detection sensitivity of molecules using nuclear magnetic resonance (NMR)/magnetic resonance imaging (MRI). This methodology enables real-time imaging of dynamic metabolic status in vivo using MRI. To expand the targetable metabolic reactions, there is a demand for developing exogenous, i.e., artificially designed, DNP-NMR molecular probes; however, complying with the requirements of practical DNP-NMR molecular probes is challenging because of the lack of established design guidelines. Here, we report Ala-[1-13C]Gly-d2-NMe2 as a DNP-NMR molecular probe for in vivo detection of aminopeptidase N activity. We developed this probe rationally through precise structural investigation, calculation, biochemical assessment, and advanced molecular design to achieve rapid and detectable responses to enzyme activity in vivo. With the fabricated probe, we successfully detected enzymatic activity in vivo. This report presents a comprehensive approach for the development of artificially derived, practical DNP-NMR molecular probes through structure-guided molecular design.


Whole-Genome Characteristics and Polymorphic Analysis of Vietnamese Rice Landraces as a Comprehensive Information Resource for Marker-Assisted Selection.
Next generation sequencing technologies have provided numerous opportunities for application in the study of whole plant genomes. In this study, we present the sequencing and bioinformatic analyses of five typical rice landraces including three indica and two japonica with potential blast resistance. A total of 688.4 million 100 bp paired-end reads have yielded approximately 30-fold coverage to compare with the Nipponbare reference genome. Among them, a small number of reads were mapped to both chromosomes and organellar genomes. Over two million and eight hundred thousand single nucleotide polymorphisms (SNPs) and insertions and deletions (InDels) in indica and japonica lines have been determined, which potentially have significant impacts on multiple transcripts of genes. SNP deserts, contiguous SNP-low regions, were found on chromosomes 1, 4, and 5 of all genomes of rice examined. Based on the distribution of SNPs per 100 kilobase pairs, the phylogenetic relationships among the landraces have been constructed. This is the first step towards revealing several salient features of rice genomes in Vietnam and providing significant information resources to further marker-assisted selection (MAS) in rice breeding programs.


Minimally invasive surgery treatment for thoracic spine tumor removal: a mini-open, lateral approach.
Prospective registry. The objective of this study is to examine procedural and long-term outcomes of a mini-open, lateral approach for tumor removal in the thoracic spine. The majority of spinal tumors present as metastatic tumors in the thoracic spine. Conventional surgical treatments have been associated with high rates of approach-related morbidities as well as difficult working windows for complete tumor excision. Recent advances in minimally invasive techniques, particularly mini-open (minimally invasive, not endoscopic) approaches, help to reduce the morbidities of conventional procedures with comparable outcomes. Twenty-one consecutively treated patients at 2 institutions were treated between 2007 and 2009. Treatment variables, including operating time, estimated blood loss, length of hospital stay, and complications were collected, as were outcome measures, including the visual analog scale for pain and the Oswestry disability index. Twenty-one patients with thoracic spinal tumors were successfully treated with a minimally invasive lateral approach. Operating time, estimated blood loss, and length of hospital stay were 117 minutes, 291 mL, and 2.9 days, respectively. One (4.8%) perioperative complication occurred (pneumonia). Mean follow-up was 21 months. Two patients had residual tumor at last follow-up. Two patients died during the study as the result of other metastases (spine tumor was secondary). Visual analog scale improved from 7.7 to 2.9 and Oswestry disability index improved from 52.7% to 24.9% from preoperative to the last follow-up. The mini-open lateral approach described here can be performed safely and without many of the morbidities and difficulties associated with conventional and endoscopic procedures. Proper training in minimally invasive techniques and the use of direct-visualization minimally invasive retractors are required to safely and reproducibly treat these complex indications.


Epigenetic effects of inhibition of heat shock protein 90 (HSP90) in human pancreatic and colon cancer.
Silencing of tumor suppressor and DNA repair genes through methylation plays a role in cancer development, growth and response to therapy in colorectal and pancreatic cancers. Heat shock protein 90 (HSP90) regulates transcription of DNA methyltransferase enzymes (DNMT). In addition, DNMTs are client proteins of HSP90. The aim of this study is to evaluate the effects of HSP90 inhibition on DNA methylation in colorectal and pancreatic cancer cell lines. Our data shows that inhibition of HSP90 using ganetespib resulted in downregulation of mRNA and protein expression of DNMT1, DNMT3A, and DNMT3B in HT-29 and MIA PaCa-2 cell lines. This in turn was associated with a drop in the fraction of methylated cytosine residues and re-expression of silenced genes including MLH-1, P16 and SPARC. These effects were validated in HT-29 tumors implanted subcutaneously in mice following in vivo administration of ganetespib. This work demonstrates the effectiveness of ganetespib, an HSP90 inhibitor in modulating DNA methylation through downregulation of DNMT expression.


A novel splice variant of folate receptor 4 predominantly expressed in regulatory T cells.
Regulatory T cells (Tregs) are required for proper maintenance of immunological self-tolerance and immune homeostasis. Folate receptor 4 (FR4) is expressed at high levels in transforming growth factor-beta (TGF-β)-induced Tregs and natural Tregs. Moreover, antibody-mediated targeting of FR4 is sufficient to mediate Treg depletion. In this study, we describe a novel FR4 transcript variant, FR4D3, in which exon 3 is deleted. The mRNA of FR4D3 encodes a FR4 variant truncated by 189 bp. FR4D3 was found to be predominantly expressed in CD4(+)CD25(+) Treg cells. Overexpression of FR4D3 in CD4(+)CD25(+) Treg cells in vitro stimulated proliferation, which may modulate the ability of these cells to bind and incorporate folic acid. Our results suggested that high levels of FR4D3 may be critical to support the substantial proliferative capacity of Treg cells.


Probing the Conformational Changes Associated with DNA Binding to PARP1.
Poly(ADP-ribose) polymerase 1 (PARP1) is an important first responder in the mechanism of DNA repair in eukaryotic cells. It is also a validated drug target, with four different PARP inhibitors (PARPi) approved for the treatment of BRCA-negative cancers. Despite past efforts, many aspects of PARPi are poorly understood, in particular their ability to trap PARP1 on chromatin and the relationships between their potencies, cellular toxicities, and trapping efficiencies. Because PARP trapping is widely believed to originate in allosteric coupling between DNA binding and the catalytic site, we further investigated the binding properties of PARP1 to a model for DNA with a double-strand break in the presence and absence of PARPi. Specifically, we have used sequential mixing stopped-flow spectroscopy to identify a slow conformational change that follows rapid DNA binding. Using a range of DNA concentrations and different mutants of PARP1 we demonstrate that this conformational change is one of the steps of the "monkey bar mechanism" that promotes DNA-dependent dissociation of DNA. This conformational change also corresponds to the previously identified conformational change associated with DNA-dependent activation of PARP1. Despite linking the conformational change associated with DNA binding and release to DNA activation, we find no evidence for PARPi perturbing this allosteric coupling.


Suicide risk profiles among service members and veterans exposed to suicide.
Rates of suicide exposure are high among service members and Veterans and are especially concerning given the link between suicide exposure and subsequent suicide risk. However, to date, it is unclear which individuals who are exposed to suicide are subsequently at high risk for suicide. Latent profile analysis (LPA) can provide information on unique risk profiles and subgroups of service members and Veterans who have higher suicide risk after suicide exposure, which has not yet been empirically studied. The purpose of this study was to utilize LPA to identify subgroups of service members and Veterans who are at the highest risk for suicidal thoughts and behaviors following suicide exposure. We analyzed data using LPA from 2570 service members and Veterans (82.1% male, 69.5% White, and 12.1% Latino/a/x) who completed the Military Suicide Research Consortium's Common Data Elements, a battery of self-report suicide-related measures. Psychopathology, substance use, mental health service utilization, interpersonal theory of suicide, and suicide exposure variables were used to validate classes. Three latent classes emerged from analyses, one low-risk class and two-high risk classes with differing profile compositions (one primarily differentiated by anxiety symptoms and one differentiated by substance use). Class-specific recommendations for suicide prevention efforts will be discussed.


Negative thoughts after childbirth: development and preliminary validation of a self-report scale.
This study describes the development and initial validation of a questionnaire that is suitable for detecting and measuring postpartum negative thoughts. Semistructured interviews with mothers who had suffered from postnatal depression were conducted to inform the content of the questionnaire. The initial questionnaire, alongside other measures, was then administered to a nonclinical sample of mothers with babies aged 0-7 months. Using principal components analysis, a two-factor structure was obtained for the Postnatal Negative Thoughts Questionnaire (PNTQ). The factors included appraisal of cognition, emotion, and situation (ACES) and baby-related and motherhood negative thoughts (BRM-NT). The psychometric properties demonstrated acceptable validity, satisfactory test-retest reliability, and internal consistency. These findings suggest that the PNTQ is a reliable and valid measure for assessing postpartum negative thoughts. Consistent with previous research, findings also suggest that appraisal of negative thoughts is more strongly related to postpartum depression than to the experience of negative thoughts per se. Clinicians may use the PNTQ to offer new mothers the opportunity to assess whether negative thoughts or metacognitive appraisals are being experienced as problematic. Additionally, a direct focus upon the metacognitive appraisals of postpartum negative thoughts may provide a useful adjunct to traditional cognitive therapy approaches. Recommendations for future research are discussed.


MACULAR ADD-ON INTRAOCULAR LENS SUCCESSFULLY RESTORES READING VISION IN EYES WITH END-STAGE DIABETIC MACULAR DISEASE.
To report the outcomes of macular add-on intraocular lens implantation in improving reading vision in patients with bilateral advanced diabetic maculopathy. In this retrospective study, a supplementary bifocal sulcus intraocular lens (Scharioth Macular Lens) was implanted in the better-seeing eye of five patients. Baseline-corrected distance vision, corrected near visual acuity, a preoperative simulation test, and multimodal imaging were collected. The primary outcome was the uncorrected near visual acuity at a working distance of 15 cm, at a 12-month follow-up. Study patients included 3 cases of refractory subfoveal exudation and 2 cases of diabetic macular ischemia. A preoperative test to assess the potential gain in near vision showed an improvement of ≥2 paragraphs on the RADNER reading chart in all patients. At 12 months, median reading vision (corrected near visual acuity at 15 cm) significantly improved from 20/125 (range 20/50-20/200) preoperatively to uncorrected near visual acuity (at 15 cm) of 20/50 (range 20/40-20/80) (P = 0.042; Wilcoxon signed-ranks test). Distance vision remained unchanged in four patients. All patients were able to achieve the size of newsprint (20/50 Snellen equivalent), within the first 3 months. The macular add-on intraocular lens improves reading vision in visually impaired patients due to end-stage diabetic macular disease.


[Evaluation of efficacy of 1 year after completing the 2-year specific sublingual immunotherapy treatment course in allergic rhinitis].
Objective: To evaluate the efficacy of treatment at 1 year after completing the 2-year specific sublingual immunotherapy in patients with allergic rhinitis (AR). Methods: A retrospective analysis of 124 patients with AR who were allergic to dust mite and treated in the First Affiliated Hospital of Wenzhou Medical University since 2012 was performed. All patients achieved sublingual immunotherapy (SLIT) for 2 years and had a regular follow-up over 1 year. Efficacy evaluation index such as VAS score, total nasal symptom score and total medication score were compared between the time of SLIT for 2 years, and 1 year after termination of SLIT. Paired t test was used for the comparison of scores. Results: Compared with those before treatment, the VAS score, total nasal symptom score and total medication score at the time of SLIT for 2 years were significantly decreased in all patients with AR [(9.40±5.96) vs (24.78±6.36), (4.53±2.06) vs (9.51±2.02), (0.42±0.87) vs (3.02±0.41), t value was 17.627, 24.600, 5.331, respectively, all P<0.01]. Compared with 2 years SLIT treatment, VAS score and total nasal symptom score were significantly increased than 1 year after termination of immunotherapy [(12.52±6.92) vs (9.40±5.96), (5.30±2.36) vs (4.53±2.06), t=5.199, 3.744, respectively, all P<0.01], but the total medication score showed no significant difference [(0.34±0.84) vs (0.42±0.87), t=-1.043, P>0.05]. Conclusions: The symptoms after 2 years SLIT in patients with AR have been well controlled. One year after the termination of SLIT, there is a certain rebound in symptoms, 2 years SLIT may not be the ideal treatment cycle for AR.


Diverse Systems for Efficient Sequence Insertion and Replacement in Precise Plant Genome Editing.
CRISPR-mediated genome editing has been widely applied in plants to make uncomplicated genomic modifications including gene knockout and base changes. However, the introduction of many genetic variants related to valuable agronomic traits requires complex and precise DNA changes. Different CRISPR systems have been developed to achieve efficient sequence insertion and replacement but with limited success. A recent study has significantly improved NHEJ- and HDR-mediated sequence insertion and replacement using chemically modified donor templates. Together with other newly developed precise editing systems, such as prime editing and CRISPR-associated transposases, these technologies will provide new avenues to further the plant genome editing field.


Improving hand hygiene compliance rates in the haemodialysis setting: more than just more hand rubs.
Haemodialysis patients are at high risk for developing healthcare-associated infections as well as acquiring multidrug-resistant microorganisms. Hand hygiene is considered to be the single most effective tool to prevent healthcare-associated infections. The number of indications and the extent of indication-specific compliance with hand rubs in the haemodialysis setting are currently unknown. We conducted a prospective, three-phase, observational intervention study on hand hygiene during haemodialysis treatments. Optimized hand hygiene standard operating procedures (SOPs) for dialysis connections (Intervention I) and disconnections (Intervention II) were compiled and implemented during two predefined intervention periods. A total of 8897 indications for hand rubs were observed throughout this study. In the course of the study, we identified an increase in the number of hand rubs performed (6-9, mean number per dialysis procedure), parallelled by a decrease in the indications for hand rubs (21-15), resulting in a significant increase of overall hand rub compliance (30-62%). The greatest improvement was seen before aseptic tasks (21-52%), the indication with the greatest impact on preventing healthcare-associated infections. There was no difference between haemodialysis via central venous catheter access or arterio-venous (AV) fistulas. This study provides the first detailed data on the number of and indications for hand rubs during dialysis. An >100% increase in overall hand hygiene compliance could be achieved by a comparably moderate increase in hand rubs performed in combination with optimized hand hygiene SOPs.


Synthesis and evaluation of new 1-oxa-8-azaspiro[4.5]decane derivatives as candidate radioligands for sigma-1 receptors.
We report the design, synthesis, and evaluation of a series of 1-oxa-8-azaspiro[4.5]decane and 1,5-dioxa-9-azaspiro[5.5]undecane derivatives as selective σ1 receptor ligands. All seven ligands exhibited nanomolar affinity for σ1 receptors (Ki(σ1) = 0.47 - 12.1 nM) and moderate selectivity over σ2 receptors (Ki(σ2)/ Ki(σ1) = 2 - 44). Compound 8, with the best selectivity among these ligands, was selected for radiolabeling and further evaluation. Radioligand [18F]8 was prepared via nucleophilic 18F-substitution of the corresponding tosylate precursor, with an overall isolated radiochemical yield of 12-35%, a radiochemical purity of greater than 99%, and molar activity of 94 - 121 GBq/μmol. Biodistribution studies of [18F]8 in mice demonstrated high initial brain uptake at 2 min. Pretreatment with SA4503 resulted in significantly reduced brain-to-blood ratio (70% - 75% at 30 min). Ex vivo autoradiography in ICR mice demonstrated high accumulation of the radiotracer in σ1 receptor-rich brain areas. These findings suggest that [18F]8 could be a lead compound for further structural modifications to develop potential brain imaging agents for σ1 receptors.


Evaluation of Probiotic Diversity from Soybean (Glycine max) Seeds and Sprouts Using Illumina-Based Sequencing Method.
There is increasing interest in the use of plant probiotics as environmental-friendly and healthy biofertilizers. The study aimed at selecting for novel probiotic candidates of soybean (Glycine max). The bacteriome and mycobiome of soybean sprouts and seeds were analyzed by Illumina-based sequencing. Seeds contained more diverse bacteria than those in sprouts. The seeds contained similar fungal diversity with sprouts. Total 15 bacterial OTUs and 4 fungal OTUs were detected in seeds and sprouts simultaneously, suggesting that the sprouts contained bacterial and fungal taxa transmitted from seeds. The Halothiobacillus was the most dominant bacterial genus observed and coexisted in seeds and sprouts. The OTUs belonged to Ascomycota were the most dominant fungal taxa observed in seeds and sprouts. Halothiobacillus was firstly identified as endophytic probiotics of soybean. The results suggested that sprouts might contain diverse plant probiotics of mature plants and Illumina-based sequencing can be used to screen for probiotic candidates.


Insights Into Emissions and Exposures From Use of Industrial-Scale Additive Manufacturing Machines.
Emerging reports suggest the potential for adverse health effects from exposure to emissions from some additive manufacturing (AM) processes. There is a paucity of real-world data on emissions from AM machines in industrial workplaces and personal exposures among AM operators. Airborne particle and organic chemical emissions and personal exposures were characterized using real-time and time-integrated sampling techniques in four manufacturing facilities using industrial-scale material extrusion and material jetting AM processes. Using a condensation nuclei counter, number-based particle emission rates (ERs) (number/min) from material extrusion AM machines ranged from 4.1 × 1010 (Ultem filament) to 2.2 × 1011 [acrylonitrile butadiene styrene and polycarbonate filaments). For these same machines, total volatile organic compound ERs (μg/min) ranged from 1.9 × 104 (acrylonitrile butadiene styrene and polycarbonate) to 9.4 × 104 (Ultem). For the material jetting machines, the number-based particle ER was higher when the lid was open (2.3 × 1010 number/min) than when the lid was closed (1.5-5.5 × 109 number/min); total volatile organic compound ERs were similar regardless of the lid position. Low levels of acetone, benzene, toluene, and m,p-xylene were common to both AM processes. Carbonyl compounds were detected; however, none were specifically attributed to the AM processes. Personal exposures to metals (aluminum and iron) and eight volatile organic compounds were all below National Institute for Occupational Safety and Health (NIOSH)-recommended exposure levels. Industrial-scale AM machines using thermoplastics and resins released particles and organic vapors into workplace air. More research is needed to understand factors influencing real-world industrial-scale AM process emissions and exposures.


Obstetric anaesthetists' workload.
The current recommendation of one consultant session per 500 deliveries with full sessional cover for units over 3000 deliveries is arbitrary and is not based on workload. The Audit Commission has questioned the wide variability of anaesthetic staffing on labour wards. The aim of this study was to investigate whether there is a relationship between current workload and obstetric anaesthetic staffing in five maternity units. In 1998, a 2-week diary of workload in the participating obstetric units was assessed in terms of staffing and clinical and nonclinical activities. The busiest time was 08:00 to 12:30. The working patterns between consultants and trainees varied considerably. Time spent on nonclinical activity by consultants averaged 51%. Any calculation of consultant sessions will need to take into account nonclinical activities. Other factors requiring consideration are the number of high-risk cases, as well as the number and experience of trainees.


[New Evidence of Efficacy and Safety of Rivaroxaban in Routine Clinical Practice].
The article contains results of two studies assessing efficacy and safety of rivaroxaban in routine clinical practice which were presented at European Congress of Cardiology 2015. These studies (recently completed XANTUS and ongoing post-marketing safety surveillance study in the United States) in total included more than 45 000 patients from Europe, Russia, Canada and the United States.


How do clinicians search for and access biomedical literature to answer clinical questions?
This paper presents a retrospective data analysis on how 75 clinicians searched for and accessed biomedical literature from an online information retrieval system to answer six clinical scenarios. Using likelihood ratio measures to quantify the impact of documents on a decision, and a graphical representation to model clinicians' journeys of accessing documents, this analysis reveals that clinicians did not necessarily arrive at the same answer after having accessed the same document, and that documents did not influence clinicians in the same manner. A possible explanation for these phenomena is that people experience cognitive biases during information searching which influence their decision outcome. This analysis raises the hypotheses that people experience the anchoring effect, order effects, exposure effect and reinforcement effect while searching for information and that these biases may subsequently influence the way decisions are made.


Theoretical analysis of catalytic-sRNA-mediated gene silencing.
Small regulatory RNA (sRNA) that acts by an antisense mechanism is critical for gene regulation at the posttranscriptional level. Recently, an Hfq-dependent sRNA named MicM, which is related to the regulation of outer membrane protein, was verified as a novel antisense sRNA due to its catalytic mode of regulation. Here we propose a simple kinetic model for the enzyme-like regulation mode of sRNA and study in detail the noise properties of the target gene under various recycling rates of the regulator. We predict that the recycling rate of sRNA and other relative parameters have significant influence on the noise strength of target expression. In comparison with the stoichiometric regulatory mode, a lesser fluctuation of target expression was observed near the threshold at which the transcription rates of both sRNA and target mRNA equal each other. We also found that the new mode is better in terms of rapid response to external signals. However, it needs more time to achieve target recovery if the stimulating signal disappears. Additionally, the obtained time evolution results of the MicM-ybfM interaction system based on our model are consistent with previous experimental results, serving as experimental evidence to back up our theoretical analysis.


Glaucomatous changes in lamina pores shape within the lamina cribrosa using wide bandwidth, femtosecond mode-locked laser OCT.
The lamina cribrosa (LC) is known to play a critical role in the pathogenesis of glaucoma. Although it has been reported that striae-shaped or slit-shaped lamina pores are more frequent in eyes with primary open angle glaucoma (POAG), this observation is based only on fundus photography. The primary object of this study is to perform layer-by-layer comparisons of the shape of lamina pores within the LC in vivo. Cross-sectional study. Optic nerve head B-scans were obtained using custom-made broad-wavelength optical coherence tomography with a mode-locked laser. A total of 300 single B-scans per eye were obtained and three-dimensional images were rendered from these image sequences to obtain 2-μm thin-slice en face images of the LC. Elongation indices (EIs) of the lamina pores were measured from the anterior surface (AS) of the LC to the deeper layers in 40-μm increments. Thirteen eyes from 10 primary open angle glaucoma (POAG) patients of mean deviation -15.2 (-16.5, -12.9) (median [25,75 percentile]) dB and 10 eyes from 7 normal controls were studied. Although the EI value was not significantly different between the superior, temporal and inferior regions of the LC at any depth level in either group, it was greater at the AS than at the 40 μm and 80 μm depth levels (P < .001) in both groups, and was greater in the POAG group only at the AS and 40 μm depth level (P ≤ .05). After adjustment for age and refraction, the effects of depth and presence of POAG on the EI value remained significant. Also, the severity of glaucoma and depth were significant factors associated with EI in multivariate analysis. Elongation of lamina pores was significantly more evident at the anterior surface and the 40-μm depth level of the LC in POAG eyes than in normal eyes, suggesting that nerve fiber bundles passing through the LC were under greater stress in the anterior layers of the LC.


Problematic eating behaviour in Turkish children aged 12-72 months: characteristics of mothers and children.
The aim of this study was to determine prevalence of problematic eating behaviour (PEB), associated risk factors, feeding practices including place of meal, variety of diet, and habits of consuming junk food, the mothers' perception of the child growth status in comparison to his/ her peers, and the effects on anthropometric measurements. This study was carried out among children aged 12-72 months who attended the outpatient clinic in the Ihsan Dogramaci Children's Hospital between February-June 2007. Three hundred and thirty-one mothers of children were asked to complete an extensive questionnaire covering socio-demographic characteristics and their child's general eating behaviour and feeding practices at mealtimes. Children with PEB were identified based on their mothers' statements. Three hundred and thirty-one cases were 3.32 +/- 1.39 years old. One hundred thirty-five mothers reported having a child with PEB. The mothers described the children's problematic behaviour as: need to walk around with the child during mealtime (45.6%), watching TV during meals (41.9%), picky or fussy eating (39%), vomiting and/or retching (25.7%), retaining food in the mouth for a long time (20.6%), and not eating solid foods (11.8%). In children who had ate neither meat nor vegetables and fruits, took cod-liver oil-containing supplement during the course of the study, and had taken iron supplements in the first year of life, PEB was more frequent than in others. The mean z scores of weight for age (WAZ) were significantly lower in cases with PEB than without PEB. Counselling and supporting of the mother/caregiver could alleviate the effect of inappropriate solutions taken by families. Insistence on composing of the diet variety including especially vegetables, fruits and meat may be promoted by provision of alternative cooking/presentation samples to mothers of children who refuse some foods. Pediatricians should be alerted that lower WAZ values may be a warning indicating a problem which may cause stagnated growth in children with PEB.


The effects of rigidity of internal fixation plates on long bone remodeling. A biomechanical and quantitative histological study.
The effect of rigidity of internal fixation plates on long bone remodeling was studied using two types of plates with considerable differences in stiffness. The plated bones were subjected to bioengineering quantitative histological, and cortical thickness studies after 9 and 12 months. The biomechanical results, together with the quantitative histological measurements of the macroscopic architecture, showed that tissue characteristics of the plated bones were similar. However, because of the larger cortical area, the less regidly plated femora can sustain significantly higher loads and energy before failure. Cortical thickness measurements also showed that rigid plate immobilization results in thinning of the cortex of the underlying bone. The experimental results suggest that cortical bone remodels according to functional stress demands, and the osteoporosis secondary to rigid plate protection is consequent to thinking of its cortex.


Association between body condition score change during the dry period and postpartum health and performance.
The objectives of the current study were to determine the association between body condition score change during the dry period (ΔBCS) and postpartum health and reproductive and productive performance of Holstein cows. Data from 16,104 lactations from 9,950 parous cows from 2 dairies located in the San Joaquin Valley of California were used. Within dairy, cows were scored for body condition at dry off and parturition by the same herd workers, who were trained by veterinarians from the Veterinary Medicine Teaching and Research Center of the University of California Davis. Cows were classified as having excessive loss of BCS (ΔBCS ≤-0.75; n = 1,604), moderate loss of BCS (ΔBCS = -0.5 to -0.25; n = 6,430), no change in BCS (ΔBCS = 0; n = 4,819), and gained BCS (ΔBCS ≥0.25; n = 3,251). Data regarding morbidity, mortality, and reproductive and productive performance were recorded until 305 d postpartum or until cows were dried off or left the herd. Loss of BCS during the dry period was associated with greater incidence of uterine disease and indigestion. Additionally, loss of BCS during the dry period was associated with greater likelihood of treatment with antimicrobials, anti-inflammatories, and supportive therapy. Loss of BCS during the dry period was associated with reduced likelihood of pregnancy after the first and second postpartum inseminations. Cows that gained BCS during the dry period had greater yield of milk, fat, and protein and had reduced somatic cell linear score in the subsequent lactation. In the current study, loss of BCS during the dry period was a predisposing factor associated with health disorders and reduced productive and reproductive performance in Holstein cows.


D-Aspartate Upregulates DAAM1 Protein Levels in the Rat Testis and Induces Its Localization in Spermatogonia Nucleus.
Cell differentiation during spermatogenesis requires a proper actin dynamic, regulated by several proteins, including formins. Disheveled-Associated-Activator of Morphogenesis1 (DAAM1) belongs to the formins and promotes actin polymerization. Our results showed that oral D-Aspartate (D-Asp) administration, an excitatory amino acid, increased DAAM1 protein levels in germ cells cytoplasm of rat testis. Interestingly, after the treatment, DAAM1 also localized in rat spermatogonia (SPG) and mouse GC-1 cells nuclei. We provided bioinformatic evidence that DAAM1 sequence has two predicted NLS, supporting its nuclear localization. The data also suggested a role of D-Asp in promoting DAAM1 shuttling to the nuclear compartment of those proliferative cells. In addition, the proliferative action induced by D-Asp is confirmed by the increased levels of PCNA, a protein expressed in the nucleus of cells in the S phase and p-H3, a histone crucial for chromatin condensation during mitosis and meiosis. In conclusion, we demonstrated, for the first time, an increased DAAM1 protein levels following D-Asp treatment in rat testis and also its localization in the nucleus of rat SPG and in mouse GC-1 cells. Our results suggest an assumed role for this formin as a regulator of actin dynamics in both cytoplasm and nuclei of the germ cells.


Methylation and expression of p16INK4 tumor suppressor gene in primary colorectal cancer tissues.
It is known that p16(INK4) tumor suppressor gene expression in colon cancer cells is repressed by methylation at the CpG island of promoter, but in vivo silencing of p16 gene is not fully understood. Some studies showed that primary colorectal cancer (CRC) tissues often overexpress the p16 protein, while others showed the high incidence of p16 methylation. The aim of this study was to clarify p16 gene regulation in vivo. We used real-time methylation-specific PCR (MSP) to examine density of p16 methylation, and immunohistochemistry, Western blot analysis to determine p16 protein expression. Methylation was detected in 5 CRC cell lines tested and 9 of 21 (42.9%) CRCs. Four of 5 CRC cell lines did not express p16 mRNA, but 6 of 9 CRCs did express p16 mRNA even with methylation. Real-time MSP showed that CRC tissues had a wide variety in methylation density (methylation index: 0.28-0.91) and that highly methylated CRC tissues displayed significantly lower p16 mRNA expression than those with no-methylation or low-methylation. Immunohistochemistry showed that the majority of CRCs (53 of 55: 96.4%) overexpressed the p16 protein. Low p16 expression was associated with lymph node metastasis (p=0.003) and large tumor size (p=0.048). Western blot in a subset of non-tumor and tumor samples showed a consistent overexpression of the p16 protein. These results showed that CRC tissues displayed variable methylation density, which may be characteristics of p16 gene methylation in vivo. Our data suggest that a low p16 expression due to methylation may contribute to tumor enlargement and expansion of CRC.


Recombinant AfusinC, an anionic fungal CSαβ defensin from Aspergillus fumigatus, exhibits antimicrobial activity against gram-positive bacteria.
Antimicrobial peptides (AMPs) are short and generally positively charged peptides found in a wide variety of organisms. CSαβ defensins are a group of AMPs. These defensins are composed of an α-helix and a β-sheet linked by three or four disulphide bridges. In this study, we describe the antimicrobial activity of an anionic CSαβ fungal defensin from Aspergillus fumigatus, AfusinC. AfusinC was recombinantly produced as a fusion protein in Escherichia coli. The tag was removed by proteolytic cleavage, and AfusinC was purified by size exclusion chromatography. About 0.8 mg of recombinant AfusinC was obtained from 1 L of culture. Recombinant AfusinC was active against mainly gram-positive bacteria including human pathogens and a multiresistant-strain of A. aureus. Additionally, AfusinC showed bactericidal effect against Micrococcus luteus.


Scandinavian total ankle replacement: a 3.7-year average followup of 65 patients.
The purpose of the current prospective study was to determine the midterm results of 68 total ankle replacements with the Scandinavian Total Ankle Replacement (S.T.A.R.) prosthesis. The 65 patients (34 women and 31 men; mean age at surgery, 56.1 years [range, 22-85 years]) were assessed clinically and radiologically after 3.7 years (range, 2.4-6.2 years). Thirty-five patients (54%) were totally pain-free. The overall clinical score was graded as excellent or good in 67 ankles. The American Orthopaedic Foot and Ankle Society hindfoot score improved from 24.7 points (range, 3-44 points) preoperatively to 84.3 points (range, 44-100 points) at followup. Three patients (three ankles, 4.4%) had a ballooning bone lysis on the tibial side. Despite prophylaxis, periarticular hypertrophic bone formation was seen in 43 ankles (63%; 42 patients), associated with a decrease of dorsiflexion and plantar flexion. Nine ankles (13%; nine patients) had revision surgery because of problems with the components and 14 ankles (21%; 14 patients) had secondary or additional operations. All revision or secondary surgeries were successful, and no ankle had to be converted to an ankle arthrodesis. The early experience with the S.T.A.R. ankle implant is encouraging, although we have encountered more complications and potential problems than previously reported.


Lovastatin and perillyl alcohol inhibit glioma cell invasion, migration, and proliferation--impact of Ras-/Rho-prenylation.
Alterations in small GTPase mediated signal transduction pathways have emerged as a central step in the molecular pathogenesis of glioblastoma (GBM), the most common malignant brain tumor in adults. Farnesylpyrophosphate (FPP) and geranylgeranylpyrophosphate (GGPP) are derived from mevalonate, whose production is catalyzed by 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase. Prenylation by FPP and GGPP is required for membrane insertion and oncogenic function of Ras- and Rho-proteins, within the stimulation of the Ras-Raf-MEK-ERK pathway. A straightforward prediction from HMG-CoA reductase inhibitor studies is that statins decrease FPP and GGPP levels and diminish ERK signaling ensuring less proliferation and migration of cancer cells. Perillyl alcohol (POH), a naturally occurring monoterpene inhibits prenyltransferases and is able to inhibit cancer cell growth, but the underlying mechanism is still unclear. We here report that lovastatin (LOV) and POH impair the regulation of the mevalonate- and the Ras-Raf-MEK-ERK pathway in U87 and U343 glioblastoma cells. Both compounds affected the post-translational modification of H-Ras and Rac1. While LOV diminished the substrates of the transferase reaction that catalyze prenylation, POH inhibited the enzymes itself. Our data highlight the impact of isoprenoids for post-translational modification of small GTPases promoting proliferation, migration and invasion capabilities in glioma cells.


Cyclin mRNA stability does not vary during the cell cycle.
The cyclins are tightly regulated elements governing eukaryotic cell cycle progression by means of sequential activation-inactivation of cyclin-dependent kinases. In one manifestation of this regulation, the mRNA levels of several cyclin genes oscillate during the cycle in mammalian cells. Such cycle-dependent fluctuations in transcript levels could result from changes not only in rates of transcription, but also in mRNA stability. Here we used a new, minimally-disturbing method for producing multi-cycle synchronous growth of human MOLT-4 cells, in combination with quantitative real-time RT-PCR, to compare cell cycle-dependent transcript levels and half-lives of cyclin A2, B1, D3, E and PCNA mRNAs. While all mRNA levels except cyclin D3 varied in the cycle, there were no apparent variations in message half-lives. This differs from several previous reports of dramatic fluctuations in the stabilities of cyclin mRNAs, and infers that fluctuations in cyclin mRNA transcript levels during the MOLT-4 cell cycle are not due to variations in half-lives. The discrepancy in mRNA stability determinations could be due to differences in cell types or synchronization methods, but our findings may be representative of mRNA processing in the cycle of cells in unstressed steady-state growth.


Substance P and bombesin elevate cytosolic Ca2+ by different molecular mechanisms in a rat pancreatic acinar cell line.
1. Dual-excitation microfluorometry (Fura-2 as indicator) was employed to monitor directly changes in the cytosolic calcium concentration [( Ca2+]i) in single cells. We investigated and compared the effects of stimulation of AR42J rat pancreatic acinar cells by two peptide agonists, substance P and bombesin. 2. Substance P (10(-7) M) and bombesin (10(-8) M) each gave rise to a marked, but transient, elevation in [Ca2+]i. The calcium signals evoked by the two peptides were qualitatively and quantitatively very similar. However, in the absence of extracellular Ca2+ the response to substance P, but not bombesin, was abolished. These results suggest that substance P induces calcium influx across the cell surface membrane but does not release calcium from internal stores. Bombesin in marked contrast releases calcium from intracellular stores in the absence of any detectable calcium influx. 3. Depolarization by high-K+ extracellular solutions evoked a marked, but transient, rise in [Ca2+]i. This elevation in [Ca2+]i was strictly dependent upon the presence of Ca2+ in extracellular media. 4. Nifedipine (5 x 10(-6) M), an antagonist of L-type voltage-dependent Ca2+ channels, blocked the elevations in [Ca2+]i induced by either substance P or high-K+ solutions, but not that evoked by application of bombesin. 5. Patch-clamp, single-channel current recordings from cell-attached patches of membrane confirmed the presence of voltage-dependent calcium channels in the surface membranes of AR42J cells. Whole-cell current recordings demonstrated voltage-dependent inward Ca2+ (Ba2+) currents which were increased in amplitude by substance P and blocked by nifedipine. 6. The protein kinase C (PKC) activators, the phorbol diester, phorbol 1,2-myristate 13-acetate (PMA, 10(-7) M), and cell-permeable diacylglycerol analogues, 1-oleoyl-2-acetyl-sn-glycerol (OAG, 2.5 x 10(-6) M) and sn-2-dioctanoyl glycerol (DiC8, 2.5 x 10(-6) M), mimicked the effect of substance P, but not bombesin, in elevating [Ca2+]i in a manner that was blocked by removal of extracellular Ca2+ or application of nifedipine. 7. The PKC inhibitor, polymyxin B (2.5 x 10(-6) M), applied 2 min prior to stimulation blocked the effects of substance P and PKC activators, but not bombesin, in elevating [Ca2+]i. 8. The calcium signals evoked by substance P and bombesin are achieved by activation of different molecular mechanisms. Substance P, the evidence suggests, activates PKC which in turn stimulates calcium influx by opening voltage-dependent Ca2+ channels in the cell surface membranes.(ABSTRACT TRUNCATED AT 400 WORDS)


Prediction of gestational hypertension by cosinor analysis of second trimester blood pressure.
We evaluated the efficacy of cosinor analysis of blood pressure (BP) at the second trimester in predicting the subsequent development of gestational hypertension. We performed the cosinor analysis of BP in 313 normotensive non-proteinuric nulliparous women at the second trimester. We followed the pregnancy up to delivery and recorded the subsequent occurrence of gestational hypertension (sGH). We compared data from this group of 33 patients with those from the 280 women with uncomplicated normotensive pregnancy. We used the ANOVA to compare the means, statistical significance was assessed at p < .05. Diastolic MESOR of women with uncomplicated pregnancy was significantly lower than the GH patients, although both groups were definitely normotensive at the time of the recording. (Diastolic: Controls M = 62.2 +/- 1.5, A = 5.1 +/- 0.7, O = 14:07, p = .0001; sGH M = 73.5 +/- 6.2m, A = 5.9 +/- 1.1, O = 14:48, p = .003; Systolic: Controls M = 108.3 +/- 2.2, A = 5.2 +/- 0.7, O = 15:00, p = .0001; sGH M = 122.4 +/- 2.1, A = 4.1 +/- 0.8, O = 15:59, p = .01) We did not find any other significant difference among the population-mean cosinors. The most sensitive cutoff of diastolic blood pressure MESOR proved to be 68 mmHg.


Advances in thickness measurements and dynamic visualization of the tear film using non-invasive optical approaches.
The thickness of tear film has been investigated under both invasive and non-invasive methods. While invasive methods are largely historical, more recent noninvasive methods are generally based on optical approaches that provide accurate, precise, and rapid measures. Optical microscopy, interferometry, and optical coherence tomography (OCT) have been developed to characterize the thickness of tear film or certain aspects of the tear film (e.g., the lipid layer). This review provides an in-depth overview on contemporary optical techniques used in studying the tear film, including both advantages and limitations of these approaches. It is anticipated that further developments of high-resolution OCT and other interferometric methods will enable a more accurate and precise measurement of the thickness of the tear film and its related dynamic properties.


Analysis of pre- and intraoperative clinical for successful operating room extubation after living donor liver transplantation: a retrospective observational cohort study.
Early extubation after liver transplantation is safe and accelerates patient recovery. Patients with end-stage liver disease undergo sarcopenic changes, and sarcopenia is associated with postoperative morbidity and mortality. We investigated the impact of core muscle mass on the feasibility of immediate extubation in the operating room (OR) after living donor liver transplantation (LDLT). A total of 295 male adult LDLT patients were retrospectively reviewed between January 2011 and December 2017. In total, 40 patients were excluded due to emergency surgery or severe encephalopathy. A total of 255 male LDLT patients were analyzed in this study. According to the OR extubation criteria, the study population was classified into immediate and conventional extubation groups (39.6 vs. 60.4%). Psoas muscle area was estimated using abdominal computed tomography and normalized by height squared (psoas muscle index [PMI]). There were no significant differences in OR extubation rates among the five attending transplant anesthesiologists. The preoperative PMI correlated with respiratory performance. The preoperative PMI was higher in the immediate extubation group than in the conventional extubation group. Potentially significant perioperative factors in the univariate analysis were entered into a multivariate analysis, in which preoperative PMI and intraoperative factors (i.e., continuous renal replacement therapy, significant post-reperfusion syndrome, and fresh frozen plasma transfusion) were associated with OR extubation. The duration of ventilator support and length of intensive care unit stay were shorter in the immediate extubation group than in the conventional extubation group, and the incidence of pneumonia and early allograft dysfunction were also lower in the immediate extubation group. Our study could improve the accuracy of predictions concerning immediate post-transplant extubation in the OR by introducing preoperative PMI into predictive models for patients who underwent elective LDLT.


Pathogen-Reduced Platelets for the Prevention of Bleeding in People of Any Age.
Are pathogen-reduced platelets as effective as standard platelets for the prevention of bleeding in people of any age requiring platelet transfusions? In people with cancer who have a low platelet count due to their disease or its treatment, pathogen-reduced platelet transfusions increase risk of platelet refractoriness and platelet transfusion requirement (high-quality evidence) but probably do not effect all-cause mortality, risk of clinically significant or severe bleeding, or risk of a serious adverse event (moderate-quality evidence). There was insufficient evidence for people with other diagnoses.


End-stage renal interstitial fibrosis in an adult ten years after ifosfamide therapy.
A 33-year-old male presented with end-stage renal failure. Renal biopsy showed severe interstitial fibrosis without glomerulopathy or vasculopathy. More than 10 years previously the patient had been successfully treated for recurrent rhabdomyosarcoma. The treatment included ifosfamide, a drug known to cause acute tubular dysfunction. Though a possible synergistic effect of previous radiation which was well within accepted tolerance limits cannot be excluded, it would appear that ifosfamide was almost certainly the major cause of the late onset chronic renal disease.


Vitamin K deficiency bleeding and early infant male circumcision in Africa.
Early infant (1-60 days of life) male circumcision is being trialed in Africa as a human immunodeficiency virus prevention strategy. Postcircumcision bleeding is particularly concerning where most infants are breastfed, and thus these infants are at increased risk of vitamin K deficiency bleeding. During a circumcision trial, one infant bled for 90 minutes postprocedure. After discovering he had not received standard prophylactic vitamin K, we gave 2 mg phytomenadione (vitamin K1) intramuscularly; bleeding stopped within 30 minutes. Vitamin K's extremely rapid action is not commonly appreciated. Neonatal vitamin K has been shown to be cost-effective. To increase availability and promote awareness of its importance, especially in low-resource settings where blood products and transfusions are limited, vitamin K should be included in the World Health Organization's Model List of Essential Medicines for Children.


Rare disease diagnosis: A review of web search, social media and large-scale data-mining approaches.
Physicians and the general public are increasingly using web-based tools to find answers to medical questions. The field of rare diseases is especially challenging and important as shown by the long delay and many mistakes associated with diagnoses. In this paper we review recent initiatives on the use of web search, social media and data mining in data repositories for medical diagnosis. We compare the retrieval accuracy on 56 rare disease cases with known diagnosis for the web search tools google.com, pubmed.gov, omim.org and our own search tool findzebra.com. We give a detailed description of IBM's Watson system and make a rough comparison between findzebra.com and Watson on subsets of the Doctor's dilemma dataset. The recall@10 and recall@20 (fraction of cases where the correct result appears in top 10 and top 20) for the 56 cases are found to be be 29%, 16%, 27% and 59% and 32%, 18%, 34% and 64%, respectively. Thus, FindZebra has a significantly (p < 0.01) higher recall than the other 3 search engines. When tested under the same conditions, Watson and FindZebra showed similar recall@10 accuracy. However, the tests were performed on different subsets of Doctors dilemma questions. Advances in technology and access to high quality data have opened new possibilities for aiding the diagnostic process. Specialized search engines, data mining tools and social media are some of the areas that hold promise.


Developmental absence of the premolar teeth: dental management.
A boy aged 11 years presented with dental pain, several carious teeth and a localized area of acute necrotizing ulcerative gingivitis (ANUG). Developmental absence of the premolar teeth was notable and additional anomalies included mid-facial hypoplasia, mandibular prognathism, transposed teeth and delayed exfoliation of the deciduous teeth. These abnormalities have significant oral, dental, orthodontic and orthognathic implications.


Spanish multicenter normative studies (Neuronorma project): norms for the abbreviated Barcelona Test.
The abbreviated Barcelona Test (a-BT) is an instrument widely used in Spain and Latin American countries for general neuropsychological assessment. The purpose of the present study was to provide new norms for the a-BT as part of the Neuronorma project. The sample consisted of 346 healthy controls. Overlapping cell procedure and midpoint techniques were applied to develop the normative data. Age, education, and sex influences were studied. Results indicated that although age and education affected the score on this test, sex did not. Raw scores were transformed to age-adjusted scaled scores (SS(A)) based on percentile ranks. These SS(A) were also converted into age-education scaled scores using a linear regression model. Norms were presented on age-education scaled scores. Also, the a-BT cognitive profile was presented and should prove to be clinically useful for interpretation. These co-normed data will allow clinicians to compare scores from a-BT with all the tests included in the Neuronorma project.


The effect of different plaque control modalities on the keratinizing potential of the sulcular epithelium in monkeys.
The purpose of this study was to determine if various plaque control regimes would be effective in permitting the gingival sulcular epithelium to keratinize. Three adult Rhesus monkeys had their teeth scaled and were placed on specific plaque control regimes including various combinations of prophylaxes, topical chlorhexidine and systemic tetracycline. Over the 2-month experimental period, weekly Plaque and Gingival Indices were determined. Subgingival plaque samples were obtained during the first 2 treatment weeks and at the end of the 1st month. Block sections of individual teeth from the different experimental regimes were serially sectioned to be evaluated histologically for inflammation and keratinization. Plaque and Gingival Indices were significantly reduced by the end of the 8th week compared to the pretreatment levels for all experimental regimes, except that the regime consisting only of three prophylaxes per week failed to reduce the Gingival Index significantly. Rubber cup prophylaxes were less effective in reducing plaque and inflammation than when the prophylaxes were combined with topical chlorhexidine application or systemic tetracycline administration. However, all experimental regimes were capable of reducing the subgingival bacterial flora to a level permitting sufficient reduction in gingival inflammation so that sulcular keratinization would occur. Some degree of sulcular keratinization was observed for every tooth of all experimental regimes. All regimes caused a significant decrease in the Inflammatory Indices relative to the control values. A negative correlation was found between the Crestal Inflammatory Index and the Keratin Width and Keratin Length, while a positive correlation was apparent between the Keratin Width and Keratin Length.


Photochemical transformations of tetrabromobisphenol A and related phenols in water.
A method was developed for studies of the phototransformation at UV irradiation of aqueous solutions of tetrabromobisphenol A (TBBPA), tribromobisphenol A (TriBBPA), tetrachlorobisphenol A (TCBPA), 2,4-dichlorophenol at various pHs as well as 2-chlorophenol, 2-bromophenol, 3,4-dichlorophenol and bisphenol A at pH 11. The absorbance spectra of the compounds and the emission spectra of the light-source were determined and used to calculate disappearance quantum yields of the photochemical reactions that were taking place. No major differences between the disappearance quantum yields of TBBPA and TCBPA were observed at pH 10, while the disappearance quantum yield of TriBBPA was approximately two times higher. The rate of decomposition of TBBPA was six times higher at pH 8 than at pH 6. Identification of the degradation products of TBBPA and TriBBPA, by GC-MS analysis and by comparison to synthesised reference compounds, indicated that TBBPA and TriBBPA decompose via different mechanisms. Three isopropylphenol derivatives; 4-isopropyl-2,6-dibromophenol, 4-isopropylene-2,6-dibromophenol and 4-(2-hydroxyisopropyl)-2,6-dibromophenol, were identified as major degradation products of TBBPA while the major degradation product of TriBBPA was tentatively identified as 2-(2,4-cyclopentadienyl)-2-(3,5-dibromo-4-hydroxyphenyl)propane.


Electric control of a canted-antiferromagnetic Chern insulator.
The interplay between band topology and magnetism can give rise to exotic states of matter. For example, magnetically doped topological insulators can realize a Chern insulator that exhibits quantized Hall resistance at zero magnetic field. While prior works have focused on ferromagnetic systems, little is known about band topology and its manipulation in antiferromagnets. Here, we report that MnBi2Te4 is a rare platform for realizing a canted-antiferromagnetic (cAFM) Chern insulator with electrical control. We show that the Chern insulator state with Chern number C = 1 appears as the AFM to canted-AFM phase transition happens. The Chern insulator state is further confirmed by observing the unusual transition of the C = 1 state in the cAFM phase to the C = 2 orbital quantum Hall states in the magnetic field induced ferromagnetic phase. Near the cAFM-AFM phase boundary, we show that the dissipationless chiral edge transport can be toggled on and off by applying an electric field alone. We attribute this switching effect to the electrical field tuning of the exchange gap alignment between the top and bottom surfaces. Our work paves the way for future studies on topological cAFM spintronics and facilitates the development of proof-of-concept Chern insulator devices.


Dosing, efficacy, and safety of recombinant factor VIIa (rFVIIa) in pediatric versus adult patients: the experience of the Hemostasis and Thrombosis Research Society (HTRS) Registry (2004-2008).
The longitudinal registry of the Hemostasis and Thrombosis Research Society (HTRS) provided data on treatment of bleeding episodes for pediatric and adult patients with congenital hemophilia complicated by inhibitors to factor VIII or IX. Data from 2,041 recombinant factor VIIa (rFVIIa)-treated episodes were recorded in the HTRS Registry between January 2004 and November 2008 and included in this retrospective analysis. The authors compared dosing, efficacy, and safety in 1,712 episodes in 284 children (age ≤18 years) and 329 episodes in 145 adults (>18 years). Patients were predominantly non-Hispanic white (52% of children, 68% of adults) with hemophilia A (91%). The majority of bleeds were spontaneous (55% pediatric, 69% adult) and occurred in the joint (55% pediatric, 65% adult). rFVIIa was used first line in most episodes (90% pediatric, 93% adult); 27% of episodes in both age groups were treated with a single dose. Children and adults received comparable median number of doses (three) and duration of therapy (1 day); adults had lower median [range] initial doses (90 [46-400] µg/kg vs. 120 [37-400] µg/kg) and median [range] total doses (270 [46-21,392] µg/kg vs. 480 [50-43,200] µg/kg) than children. Effectiveness was comparable between the groups. No thromboembolic complications were reported. Dosing of rFVIIa was somewhat higher in pediatric than adult patients in the HTRS registry population, with similar effectiveness. Safety assessment, demonstrating lack of thromboembolic events associated with treatment of bleeding episodes, was similar between the children and adults in this retrospective registry.


Liquid-chromatographic assay of urinary 5-hydroxy-3-indoleacetic acid, with electrochemical detection.
After extraction with two organic solvent mixtures, urinary 5-hydroxy-3-indoleacetic acid can be assayed by "high-performance" liquid chromatography on a reversed-phase column, with electrochemical detection. Compared to the nitrosonaphthol method (J. Biol. Chem. 216: 499, 1955), this method is more specific for detection of patients with carcinoid tumors.


Genetic dissection of reproductive performance of dairy cows under heat stress.
Heat stress negatively impacts the reproductive performance of dairy cows. The main objective of this study was to dissect the genetic basis underlying dairy cow fertility under heat stress conditions. Our first goal was to estimate genetic components of cow conception across lactations considering heat stress. Our second goal was to reveal individual genes and functional gene-sets that explain a cow's ability to conceive under thermal stress. Data consisted of 74 221 insemination records on 13 704 Holstein cows. Multitrait linear repeatability test-day models with random regressions on a function of temperature-humidity index values were used for the analyses. Heritability estimates for cow conception under heat stress were around 2-3%, whereas genetic correlations between general and thermotolerance additive genetic effects were negative and ranged between -0.35 and -0.82, indicating an unfavorable relationship between cows' ability to conceive under thermo-neutral vs. thermo-stress conditions. Whole-genome scans identified at least six genomic regions on BTA1, BTA10, BTA11, BTA17, BTA21 and BTA23 associated with conception under thermal stress. These regions harbor candidate genes such as BRWD1, EXD2, ADAM20, EPAS1, TAOK3, and NOS1, which are directly implicated in reproductive functions and cellular response to heat stress. The gene-set enrichment analysis revealed functional terms related to fertilization, developmental biology, heat shock proteins and oxidative stress, among others. Overall, our findings contribute to a better understanding of the genetics underlying the reproductive performance of dairy cattle under heat stress conditions and point out novel genomic strategies for improving thermotolerance and fertility via marker-assisted breeding.


The emerging science of precision medicine and pharmacogenomics for Parkinson's disease.
Current therapies for Parkinson's disease are problematic because they are symptomatic and have adverse effects. New drugs have failed in clinical trials because of inadequate efficacy. At the core of the problem is trying to make one drug work for all Parkinson's disease patients, when we know this premise is wrong because (1) Parkinson's disease is not a single disease, and (2) no two individuals have the same biological makeup. Precision medicine is the goal to strive for, but we are only at the beginning stages of building the infrastructure for one of the most complex projects in the history of science, and it will be a long time before Parkinson's disease reaps the benefits. Pharmacogenomics, a cornerstone of precision medicine, has already proven successful for many conditions and could also propel drug discovery and improve treatment for Parkinson's disease. To make progress in the pharmacogenomics of Parkinson's disease, we need to change course from small inconclusive candidate gene studies to large-scale rigorously planned genome-wide studies that capture the nuclear genome and the microbiome. Pharmacogenomic studies must use homogenous subtypes of Parkinson's disease or apply the brute force of statistical power to overcome heterogeneity, which will require large sample sizes achievable only via internet-based methods and electronic databases. Large-scale pharmacogenomic studies, together with biomarker discovery efforts, will yield the knowledge necessary to design clinical trials with precision to alleviate confounding by disease heterogeneity and interindividual variability in drug response, two of the major impediments to successful drug discovery and effective treatment. © 2017 International Parkinson and Movement Disorder Society.


Differences and similarities between arrhythmogenic right ventricular cardiomyopathy and athlete's heart adaptations.
Regular intensive physical activity is associated with non-pathological changes in cardiac morphology. Differential diagnosis with arrhythmogenic right ventricular cardiomyopathy (ARVC) constitutes a frequent problem, especially in athletes showing ventricular arrhythmias with left bundle branch block morphology. To assess the different clinical and non-invasive instrumental features of the subjects affected by ARVC and by athletes. Three groups of subjects (40 ARVC patients, 40 athletes and 40 controls, mean age 27 (9) years) were examined with family and personal history, physical examination, 12-lead ECG, 24-h ECG, signal-averaged ECG and 2-D and Doppler echocardiography. 12-Lead ECG was abnormal in 62% of ARVC patients versus 7.5% of athletes and 2.5% of controls (p<0.0001). Ventricular arrhythmias and late potentials were present in 70% and 55% of ARVC subjects, respectively (vs 5% of athletes and 7.5% of controls, p<0.0001). Left ventricular parietal wall thickness and left ventricular end-diastolic diameters were significantly higher in athletes. Both athletes and ARVC patients presented a right ventricular (RV) enlargement compared with controls. Moreover, RV outflow tract, measured on parasternal long axis and at the level of aortic root, was significantly larger in ARVC patients (33.6 (4.7) mm vs 29.1 (3.4) mm and 35.6 (6.8) mm vs 30.1 (2.9) mm; p<0.0001), and RV fractional shortening and ejection fraction were significantly lower in ARVC patients compared with athletes (40 (7.9)% vs 44 (10)%; p=0.05 and 52.9 (8)% vs 59.9 (4.5)%; p<0.0001). A thickened moderator band was found to be present in similar percentage in ARVC patients and athletes. An accurate clinical and instrumental non-invasive evaluation including echocardiography as imaging technique allows to distinguish RV alterations typical of ARVC from those detected in athletes as a consequence of intensive physical activity.


Cytotoxic T lymphocytes produce immune interferon in response to antigen or mitogen.
Interferon (IFN) was found to be secreted by cloned lines of murine cytotoxic T lymphocytes in response to mitogenic or antigen specific stimulation. The IFN produced was shown to be of the immune type based on its sensitivity to pH 2.0 and on the ability of an antiserum to immune IFN to neutralize the antiviral activity.


Efficient way to convert propagating waves into guided waves via gradient wire structures.
We propose a method for the design of gradient wire structures that are capable of converting propagating waves into guided waves along the wire. The conversion process is achieved by imposing an additional wave vector to the scattered waves via the gradient wire structure, such that the wave vector of scattered waves is beyond the wave number in the background medium. Thus, the scattered waves turn into evanescent waves. We demonstrate that two types of gradient wire structures, with either a gradient permittivity and a fixed radius, or a gradient radius and a fixed permittivity, can both be designed to realize such a wave conversion effect. The principle demonstrated in our work has potential applications in various areas including nanophotonics, silicone photonics, and plasmonics.


Effect of oral genistein administration in early and late phases of allergic encephalomyelitis.
Experimental allergic encephalomyelitis (EAE) is an autoimmune disease validated as animal model of multiple sclerosis (MS). Administration of genistein, a phytoestrogenic component of soy, to mice at the onset of EAE is known to attenuate the clinical signs of the disease. The potential effects of genistein on established EAE is less studied. In the current study, we aimed to compare the effects of genistein administration on EAE severity in early and late phases of the disease. The C57BL/6 mice were induced with EAE, using MOG 35-55 and gavaged with genistein (300 mg/kg) either after the appearance of the first clinical sign or 30 days post disease induction for ten days. 24 hr after the last gavage, mice were sacrificed. Brains and spleens were removed for assessing lymphocyte proliferation, cell cytotoxicity, and cytokine profile. Spinal cords were dissected to assess the amount of demyelination using Luxol fast blue/cresyl violet staining. Administering mice with genistein, after the establishment of EAE, did not reverse the clinical signs of disease. However, treating with genistein at the onset of disease alleviated the clinical signs by reducing neuronal demyelination. Genistein suppressed the production of IFN-γ and enhanced IL-10 secretion in splenocyte and brain. Genistein also reduced IL-12 and TNF-α secretion in splenocytes, suppressed the proliferation of T-cells, and reduced the cell cytotoxicity. Genistein oral therapy might only reduce EAE severity if started in early phases of the disease.


Variation of HIV infectibility of macrophages as a function of donor, stage of differentiation, and site of origin.
Heterosexual transmission of HIV-1 is likely to involve transmission of virus present in seminal fluid to inflammatory cells, particularly macrophages, present in the endometrium and peritoneal cavity. We have investigated the susceptibility of peritoneal macrophages and the corresponding autologous blood monocytes from normal women to infection by the BA-L strain of HIV-1. In 10 of 18 examples, peritoneal macrophages showed signs of infection within 4-5 days, which was earlier than the autologous monocytes. In contrast to peritoneal macrophages, lung macrophages from 10 of 11 normal donors failed to show significant reverse transcriptase (RT) values 3 weeks post infection. Monolayer cultures of monocytes cultured for 5 days prior to infection developed RT values similar overall to those of freshly isolated cells although individual donors varied as to which culture condition was optimal. The ease of infection of peritoneal macrophages did not correlate with levels of CD4 antigen or degree of pelvic inflammatory development, nor were macrophages harvested from women early in the menstrual cycle significantly more susceptible to infection than those collected from midcycle on. This unexplained heightened infectibility of peritoneal macrophages in a proportion of normal women suggests that those individuals could be more at risk for heterosexual transmission of HIV-1 infection.


Thermal Effects.
This review focuses on the research literatures published in 2014 relating to topics of thermal effects in water pollution control. This review is divided into the following sections: anaerobic wastewater and sludge treatment, biological nitrogen and phosphorus removal, membrane biological treatment, sewage sludge pyrolysis, natural treatment, resource recovery, electrolysis, oxidation and adsorption treatment.


Kampo formulations, chotosan, and yokukansan, for dementia therapy: existing clinical and preclinical evidence.
Cognitive deficits and behavioral and psychological symptoms of dementia (BPSD) are typical features of patients with dementia such as Alzheimer's disease (AD), vascular dementia (VD), and other forms of senile dementia. Clinical evidence has demonstrated the potential usefulness of chotosan (CTS) and yokukansan (YKS), traditional herbal formulations called Kampo medicines, in the treatment of cognitive disturbance and BPSD in dementia patients, although the indications targeted by CTS and YKS in Kampo medicine differ. The availability of CTS and YKS for treating dementia patients is supported by preclinical studies using animal models of dementia that include cognitive/emotional deficits caused by aging and diabetes, dementia risk factors. These studies have led not only to the concept of a neuronal basis for the CTS- and YKS-induced amelioration of cognitive function and emotional/psychiatric symptom-related behavior in animal models, but also to a proposal that ingredient(s) of Uncariae Uncis cum Ramulus, a medicinal herb included in CTS and YKS, may play an important role in the actions of these formulae in dementia patients. Further studies are needed to clarify the active ingredients of these formulae and their target endogenous molecules implicated in the anti-dementia drug-like actions.


Development and validation of an Onchocerca ochengi adult male worm gerbil model for macrofilaricidal drug screening.
Onchocerciasis currently afflicts an estimated 15 million people and is the second leading infectious cause of blindness world-wide. The development of a macrofilaricide to cure the disease has been hindered by the lack of appropriate small laboratory animal models. This study therefore, was aimed at developing and validating the Mongolian gerbil, as an Onchocerca ochengi (the closest in phylogeny to O. volvulus) adult male worm model. Mongolian gerbils (Meriones unguiculatus) were each implanted with 20 O. ochengi male worms (collected from infected cattle), in the peritoneum. Following drug or placebo treatments, the implanted worms were recovered from the animals and analyzed for burden, motility and viability. Worm recovery in control gerbils was on average 35%, with 89% of the worms being 100% motile. Treatment of the gerbils implanted with male worms with flubendazole (FBZ) resulted in a significant reduction (p = 0.0021) in worm burden (6.0% versus 27.8% in the control animals); all recovered worms from the treated group had 0% worm motility versus 91.1% motility in control animals. FBZ treatment had similar results even after four different experiments. Using this model, we tested a related drug, oxfendazole (OFZ), and found it to also significantly (p = 0.0097) affect worm motility (22.7% versus 95.0% in the control group). We have developed and validated a novel gerbil O. ochengi adult male worm model for testing new macrofilaricidal drugs in vivo. It was also used to determine the efficacy of oxfendazole in vivo.


A rapid review and narrative synthesis of hospital to community pharmacy transfer of care services in England.
A new discharge medicines service in England has been proposed for rollout in July 2020. This study aims to appraise the evidence for hospital to community pharmacy referral services in England. A rapid review methodology was adopted, findings were synthesised and reported narratively. The PubMed search engine was used and specific pharmacy journals were searched in March 2020. Studies published since 2012, reporting on transfer of care (ToC) services provided from hospitals and community pharmacies in England were included. Study data analysed included clinical outcomes; findings relating to implementation, staff and patient perceptions and experiences. Studies were assessed for risk of bias using a critical appraisal checklist. Ten studies met the inclusion criteria. The ToC services varied in relation to patients targeted, mode of referral, and post-discharge service provided. There were some conservative observations that ToC services were associated with reduced hospital readmission rates, however other outcomes were less well substantiated. Pharmacy staff were generally supportive of these services for patient benefit, however, patient engagement and accessibility issues have been reported. There is some emerging evidence that hospital to community pharmacy referral services are associated with reduced hospital readmission. Service delivery reported in these studies highlights barriers and facilitators to patient recruitment, engagement and follow-up and limitations on recording and measuring meaningful outcomes. Future work is recommended to take advantage of the implementation and delivery of the national service that includes the analysis of patient data at scale to substantiate evidence in this area.