CRF and urocortin differentially modulate GluRdelta2 expression and distribution in parallel fiber-Purkinje cell synapses.
Corticotropin-releasing factor (CRF) and urocortin (UCN) are closely related multifunctional regulators, governing, among other processes, Purkinje cell development. Here, we investigate the effects of CRF and UCN on Purkinje cells in organotypic slices. We show that both peptides upregulate delta2 ionotropic glutamate receptor gene expression, and increase the abundance of the receptor in the postsynaptic density. However, only UCN treatment results in increased delta2 protein level per Purkinje cell, implying the existence of posttranscriptional regulation of GluRdelta2 mRNA. CRF, in contrast, reduces the number of delta2-positive dendritic shafts per cell, implying that the increase of GluRdelta2 in remaining synapses may be mainly due to its retargeting. We further observed different patterns of GluRdelta2 distribution in the zone of postsynaptic density upon CRF and UCN treatment. CRF treatment results in a clustered distribution of GluRdelta2 along the postsynaptic density, whereas UCN treatment provides a linear distribution.


Food intake and body weight: effects of specific and non-specific lesions in the midbrain path of the ascending noradrenergic neurons of the rat.
Bilateral injections of 4.8 or 12 microgram of 6-hydroxydopamine hydrobromide were made in the midbrain path of the ascending norepinephrine (NE) fibers. Maximum NE depletion was obtained with the 4 microgram dose. Increasing the dose did not increase NE depletion in either the hypothalamus or the telencephalon. Increases in food intake were observed following the 12 microgram dose only. Electrolytic, copper sulfate, or 5,6-dihydroxytryptamine lesions at the same site caused increases in both food intake and body weight with only moderate (-32 to -58%) decreases in NE levels. Behavioral changes were seen in only those groups in which a large amount of non-specific tissue damage was also observed. Therefore, the behavioral effects were attributed to general tissue damage in the area and not the NE fibers specifically.


Heparan sulfate niche for cell proliferation in the adult brain.
In adulthood, new neurons and glial cells are generated from stem cells in restricted zones of the brain, namely the olfactory bulb (OB), rostral migratory stream (RMS), subventricular zone (SVZ) of the lateral ventricle, sub-callosum zone (SCZ) and sub-granular layer (SGL) of the dentate gyrus. What makes these zones germinal? We previously reported that N-sulfated heparan sulfates (N-sulfated HS) present in specialized extracellular matrix structures (fractones) and vascular basement membranes bind the neurogenic factor FGF-2 (fibroblast growth factor-2) next to stem cells in the anterior SVZ of the lateral ventricle, the most neurogenic zone in adulthood. To determine to which extent cell proliferation is associated with N-sulfated HS, we mapped N-sulfated HS and proliferating cells by immunohistochemistry throughout the adult mouse brain. We found that cell proliferation is associated with N-sulfated HS in the OB, RMS, the whole germinal SVZ, and the SCZ. Cell proliferation was weakly associated with N-sulfated HS in the SGL, but the SGL was directly connected to a sub-cortical N-sulfated HS+ extension of the meninges. The NS-sulfated HS+ structures were blood vessels in the OB, RMS and SCZ, and primarily fractones in the SVZ. N-sulfated HS+ fractones, blood vessels and meninges formed a continuum that coursed along the OB, SVZ, RMS, SCZ and SGL, challenging the view that these structures are independent germinal entities. These results support the possibility that a single anatomical system might be globally responsible for mitogenesis and ultimately the production of new neurons and glial cells in the adult brain.


A Simple Nomogram for Predicting Hospital Mortality of Patients Over 80 Years in ICU: An International Multicenter Retrospective Study.
This study aimed to develop and validate an easy-to-use intensive care unit (ICU) illness scoring system to evaluate the in-hospital mortality for very old patients (VOPs, over 80 years old). We performed a multicenter retrospective study based on the electronic ICU (eICU) Collaborative Research Database (eICU-CRD), Medical Information Mart for Intensive Care Database (MIMIC-III CareVue and MIMIC-IV), and the Amsterdam University Medical Centers Database (AmsterdamUMCdb). Least Absolute Shrinkage and Selection Operator regression was applied to variables selection. The logistic regression algorithm was used to develop the risk score and a nomogram was further generated to explain the score. We analyzed 23 704 VOPs, including 3 726 deaths (10 183 [13.5% mortality] from eICU-CRD [development set], 12 703 [17.2%] from the MIMIC, and 818 [20.8%] from the AmsterdamUMC [external validation sets]). Thirty-four variables were extracted on the first day of ICU admission, and 10 variables were finally chosen including Glasgow Coma Scale, shock index, respiratory rate, partial pressure of carbon dioxide, lactate, mechanical ventilation (yes vs no), oxygen saturation, Charlson Comorbidity Index, blood urea nitrogen, and urine output. The nomogram was developed based on the 10 variables (area under the receiver operating characteristic curve: training of 0.792, testing of 0.788, MIMIC of 0.764, and AmsterdamUMC of 0.808 [external validating]), which consistently outperformed the Sequential Organ Failure Assessment, acute physiology score III, and simplified acute physiology score II. We developed and externally validated a nomogram for predicting mortality in VOPs based on 10 commonly measured variables on the first day of ICU admission. It could be a useful tool for clinicians to identify potentially high risks of VOPs.


Vitreous opacities in infants born full-term and preterm by handheld swept-source optical coherence tomography.
To compare vitreous opacity density in infants born at term and in infants born prematurely using an investigational handheld swept-source optical coherence tomography (SS-OCT). Infants born at term underwent imaging once between 12 and 48 hours after birth; infants born prematurely were imaged at each routine retinopathy of prematurity (ROP) examination. Three masked, trained graders analyzed images. Semiautomated methods were used to quantify vitreous opacity density, which was correlated with ROP severity based on indirect ophthalmoscopy, other SS-OCT findings, and medical comorbidities. Between April 2018 and June 2019, 251 SS-OCT imaging sessions were performed on 78 infants (49% female; 36% preterm, with mean birth weight of 1018 ± 338 g and gestational age of 28.6 ± 3.2 weeks). All SS-OCT sessions produced images of adequate quality. Punctate vitreous opacities were present in 25 of 28 term infants (89%) and 41 of 50 premature infants (82%). Dice coefficient and F1 scores for intergrader agreement were 0.99 ± 0.03 and 0.77 ± 0.31, respectively. Vitreous opacity density was 0.118 ± 0.187 in prematurely born infants and 0.031 ± 0.118 in infants born at term (P = 0.009). In the former, vitreous opacity density was associated with ROP zone (P = 0.044) and stage (P = 0.031), intraventricular hemorrhage (P = 0.028), subchorionic hemorrhage (P = 0.026), and African American race (P = 0.023). In the latter, vitreous opacity density was associated with maternal diabetes (P = 0.049). Our investigational handheld SS-OCT achieved high-quality vitreoretinal images. In our study cohort, punctate vitreous opacities were a frequent finding in infants born at term and those born prematurely, with increased density in those born prematurely, particularly those with severe ROP.


Punishment inhibits an instrumental response in hooded rats.
Punishment led to the cessation of a food-rewarded bar-pressing response in hooded rats deprived of food for 23 hours. The response remained inhibited for 2 weeks, and only one of four rats resumed responding when food deprivation was increased to 47 and 71 hours.


Hall current effect in bioconvection Oldroyd-B nanofluid flow through a porous medium with Cattaneo-Christov heat and mass flux theory.
Bioconvection due to microorganisms is important area of research, considerably importance for environment and sustainable fuel cell technologies. Buongiorno nanofluid model for Cattaneo-Christov heat and mass flux theory taken into account the Oldroyd-B nanofluid and gyrotactic microorganisms in a rotating system with the effects of Hall current, and Darcy porous medium is scrutinized. The constitutive equations of the problem are transformed into nondimensional equations with the help of similarity transformations. Homotopy analysis method is used to obtain the solution. Graphs and table support the comprehesive representation of the achieved results. Radial velocity is reduced with the increasing values of relaxation time, retardation time and magnetic field parameters while heat transfer is augmented with thermal relaxation time parameter. The nanoparticles concentration is reduced with the increasing values of Schmidt number and the gyrotactic microorganisms concentration is enhanced with the increasing values of Peclet number. A nice agreement is obtained while comparing the present results numerically with the published results. The proposed mathematical model is used in biochemical engineering, meteorology, power and transportation production, optoelectronic and sensing microfabrication.


The complete chloroplast genome of Chinese medicinal herb Cynanchum chinense R. Br. (Apocynaceae) and its phylogenetic position.
Cynanchum chinense R. Br. is an indigenous Chinese medicinal herb. In this study, the complete chloroplast genome of C. chinense was reported for the first time. The genome was 158,615 bp in length, with a large single-copy region of 89,958 bp, a small single-copy region of 19,415 bp, and 2 inverted repeat regions of 24,621 bp. The genome consisted of 132 genes, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content is 37.82%. Phylogenetic analysis based on 21 complete genomes indicated that C. chinense is most closely related to Cynanchum auriculatum and Cynanchum wilfordii.


Data for molecular dynamic simulations in the OPLSAA force field: Partial charges of cholesterol, C7-hydroxycholesterol and C7-hydroperoxycholesterol, torsional parameters for the hydroperoxy group of C7-hydroperoxycholesterol.
This data article contains partial charges generated for cholesterol, C7-hydroxycholesterol and C7-hydroperoxycholesterol and torsional parameters for hydroperoxy of C7-hydroperoxycholesterol for molecular dynamics simulations in the OPLSAA force field [1] using the package Gromacs [2]. The hydroperoxy group remained unparameterized in the OPLSAA force field and the parameters obtained have the potential for re-use in similar simulations. The atom-centred point charges on each sterol molecule were derived using the restrained electrostatic potential (RESP) approach [3]. The parameters for the C7-OET-OH-HO and C8-C7-OET-OH torsion angles were derived by fitting the parameters of the torsional term (Ryckaert-Bellemans function) of the OPLSAA potential energy function to the quantum mechanical rotational energy profile calculated at CCSD(T)/cc-pVQZ level of theory. This article presents data used in the research article "Chirality affects cholesterol-oxysterol association in water, a computational study" [4].


Genetic transformation of extremophilic fungi Acidea extrema and Acidothrix acidophila.
Intact, growing cells of strongly acidophilic fungi Acidea extrema and Acidothrix acidophila have been successfully transformed by introduction of heterologous DNA fragment (composed of the glyceraldehyde-phosphate-dehydrogenase gene promoter from Emericella nidulans, a metallothionein-coding gene AsMt1 from Amanita strobiliformis and glyceraldehyde-phosphate-dehydrogenase gene terminator from Colletotrichum gloeosporioides) with the length of 1690 bp. The transformation procedure was based on the DNA transfer mediated by Agrobacterium tumefaciens bearing disarmed helper plasmid pMP90 and binary vector pCambia1300 with inserted DNA fragment of interest. The transformants proved to be mitotically stable, and the introduced gene was expressed at least at the level of transcription. Our work confirms that metabolic adaptations of strongly acidophilic fungi do not represent an obstacle for genetic transformation using conventional methods and can be potentially used for production of heterologous proteins. A promising role of the fast growing A. acidophila as active biomass in biotechnological processes is suggested not only by the low susceptibility of the culture grown at low pH to contaminations but also by reduced risk of accidental leaks of genetically modified microorganisms into the environment because highly specialized extremophilic fungi can poorly compete with common microflora under moderate conditions.


Stem-cell-based human and mouse embryo models.
Synthetic embryology aims to develop embryo-like structures from stem cells to provide new insight into early stages of mammalian development. Recent advances in synthetic embryology have highlighted the remarkable capacity of stem cells to self-organize under certain biochemical or biophysical stimulations, generating structures that recapitulate the fate and form of early mouse/human embryos, in which symmetry breaking, pattern formation, or proper morphogenesis can be observed spontaneously. Here we review recent progress on the design principles for different types of embryoids and discuss the impact of different biochemical and biophysical factors on the process of stem-cell self-organization. We also offer our thoughts about the principal future challenges.


'Worrying picture' of violence and work-related stress in staff survey.
Almost two fifths of NHS nurses in England experienced work-related stress last year, while 60% felt pressured to work while sick, according to the latest NHS staff survey.


Cingulin, a new peripheral component of tight junctions.
The tight junction (Zonula occludens), a belt-like region of contact between cells of polarized epithelia, serves as a selective barrier to small molecules and as a total barrier to large molecules, and is involved in the separation between lumenal and basolateral compartments of the epithelium. In the electron microscope, tight junctions show focal regions of apparent fusion between the adjoining cell membranes, and freeze-fractured membranes display an elaborate network of branching and anastomosing strands. Very little is known about the molecular composition and architecture of tight junctions. The first specific zonula occludens-associated protein, designated ZO-1, has recently been identified in mammalian epithelial and endothelial cells. Here we describe the identification and purification of a new component of this junctional complex in avian brush-border cells, which we name cingulin. Cingulin is an acidic, heat-stable protein, with a highly elongated shape. Immunofluorescence and immunoelectron microscopy of brush-border cells with anti-cingulin antibodies show that cingulin is localized in the apical zone of the terminal web, at the endofacial surfaces of the zonula occludens.


Quantitative differential proteomics of yeast extracellular matrix: there is more to it than meets the eye.
Saccharomyces cerevisiae multicellular communities are sustained by a scaffolding extracellular matrix, which provides spatial organization, and nutrient and water availability, and ensures group survival. According to this tissue-like biology, the yeast extracellular matrix (yECM) is analogous to the higher Eukaryotes counterpart for its polysaccharide and proteinaceous nature. Few works focused on yeast biofilms, identifying the flocculin Flo11 and several members of the HSP70 in the extracellular space. Molecular composition of the yECM, is therefore mostly unknown. The homologue of yeast Gup1 protein in high Eukaryotes (HHATL) acts as a regulator of Hedgehog signal secretion, therefore interfering in morphogenesis and cell-cell communication through the ECM, which mediates but is also regulated by this signalling pathway. In yeast, the deletion of GUP1 was associated with a vast number of diverse phenotypes including the cellular differentiation that accompanies biofilm formation. S. cerevisiae W303-1A wt strain and gup1∆ mutant were used as previously described to generate biofilm-like mats in YPDa from which the yECM proteome was extracted. The proteome from extracellular medium from batch liquid growing cultures was used as control for yECM-only secreted proteins. Proteins were separated by SDS-PAGE and 2DE. Identification was performed by HPLC, LC-MS/MS and MALDI-TOF/TOF. The protein expression comparison between the two strains was done by DIGE, and analysed by DeCyder Extended Data Analysis that included Principal Component Analysis and Hierarchical Cluster Analysis. The proteome of S. cerevisiae yECM from biofilm-like mats was purified and analysed by Nano LC-MS/MS, 2D Difference Gel Electrophoresis (DIGE), and MALDI-TOF/TOF. Two strains were compared, wild type and the mutant defective in GUP1. As controls for the identification of the yECM-only proteins, the proteome from liquid batch cultures was also identified. Proteins were grouped into distinct functional classes, mostly Metabolism, Protein Fate/Remodelling and Cell Rescue and Defence mechanisms, standing out the presence of heat shock chaperones, metalloproteinases, broad signalling cross-talkers and other putative signalling proteins. The data has been deposited to the ProteomeXchange with identifier PXD001133. yECM, as the mammalian counterpart, emerges as highly proteinaceous. As in higher Eukaryotes ECM, numerous proteins that could allow dynamic remodelling, and signalling events to occur in/and via yECM were identified. Importantly, large sets of enzymes encompassing full antagonistic metabolic pathways, suggest that mats develop into two metabolically distinct populations, suggesting that either extensive moonlighting or actual metabolism occurs extracellularly. The gup1∆ showed abnormally loose ECM texture. Accordingly, the correspondent differences in proteome unveiled acetic and citric acid producing enzymes as putative players in structural integrity maintenance.


UV-filter octyl methoxycinnamate causes reproductive toxicity associated with germline apoptosis and vitellogenin decrease in Caenorhabditis elegans.
Octyl methoxycinnamate (OMC) is a common UV filter found in personal care products such as sunscreen and cosmetics. However, OMC's presence in wastewater has raised concerns that it could potentially pollute aquatic ecosystems because of its limited biodegradability and its estrogenic disrupting properties. In this study, we investigated the environmental toxicity of OMC and its potential biomarkers using the nematode Caenorhabditis elegans. Our results showed that body length, eggs in utero, and total brood size decreased with increasing dose (experimental concentrations = 0, 1, 5, 10, 100, 500 μM for body length and eggs in utero, and 0, 5, 10 μM for total brood size) in C. elegans after L1 larval stage (the first larval stage for 0 - 12 hours post-hatching) larval stage exposure to OMC. The minimum effective concentrations were 1, 5, and 10 μM, respectively. Modeling results demonstrated that the threshold concentration of OMC inducing 10% inhibited eggs in utero was 0.33 μM (95.11 μg/L). Furthermore, germline apoptosis was induced in 10 μM OMC-treated worms (experimental concentrations = 0, 5, 10 μM). Decreased mRNA levels of vitellogenin-related genes (vit-2 and vit-6) and increased mRNA levels of apoptosis-related genes (egl-1 and ced-3) were observed in 10 μM OMC-treated C. elegans (experimental concentrations = 0, 10 μM), suggesting that reproductive toxicity was associated with decreased vitellogenin levels and germline apoptosis. In summary, our study shows that OMC is reproductively toxic and leads to reduced egg formation and decreased brood size in C. elegans by reducing vitellogenin levels and promoting germline apoptosis.


Lesson learned from the application of intersection safety devices in Edmonton.
The City of Edmonton began its intersection safety device (ISD) program in 2009 with the installation of 50 cameras throughout the city. The ISDs are approach-specific and combine automated enforcement of red-light running and speed violations during the red and green phases of the intersection control. The goal of this study is to evaluate the safety performance of ISDs within the city of Edmonton, Canada and to identify factors that can lead to successful selection of future ISD sites. A before-and-after Empirical Bayes (EB) method is used to account for regression-to-the-mean and other confounding factors. A safety performance function and yearly calibration factors are developed using data from a set of reference intersections within Edmonton. The before-and-after analysis is applied at the overall intersection level and for each approach of the ISD intersections. The results showed significant reductions that ranged from 12% to 25% for total collisions, and from 33% to 43% for angle collisions. No significant reduction was observed for severe collisions at the intersection level, however significant reductions were found at the approach level at locations with a relatively higher collision history. The impact of site selection criteria on collision reduction was also evaluated. Greater reductions were found at sites with a higher collision frequency. Additionally, the impact of intersection characteristics on collision reduction was investigated. Speed limits, presence of separated right turn lane and the number of lanes were found to impact ISD collision reduction.


Determinants of aciclovir-induced nephrotoxicity in children.
Aciclovir is the drug of choice for severe systemic herpes virus infections. Nephrotoxicity is one of the clinically significant adverse effects of this drug, but studies examining nephrotoxicity in children are scarce. To identify risk factors for aciclovir-associated nephrotoxicity in the pediatric population. A retrospective review was conducted on all children (mean age 81 months; n = 126 [74 boys]) who were treated with aciclovir in a tertiary center between July 2005 and January 2006 and who met our inclusion criteria. Glomerular filtration rate (GFR) was calculated on the first day of treatment and at the peak measured creatinine level while on therapy, using Schwartz's method. Aciclovir therapy was associated with a significant increase in serum creatinine levels and a parallel decrease in GFR (n = 93; both p <or= 0.0001). Children with immunosuppression who received a variety of other nephrotoxic drugs exhibited more severe nephrotoxicity than those not receiving nephrotoxic drugs. In multiple regression analysis, the use of nephrotoxic drugs (p = 0.02) and impaired GFR at baseline (p = 0.04) were predictive for nephrotoxicity. Within the recommended age-dependent dosage schedule of aciclovir there was no effect of dose per kg, age, or sex on nephrotoxicity. The predictors of aciclovir nephrotoxicity were the concomitant use of nephrotoxic drugs and impaired GFR at baseline.


[Use of a biocompatible antimicrobial polymer film for the prevention of incompetence of intestinal sutures in peritonitis].
The antimicrobic polymer films were used in experiment and in operations on 56 patients for strengthening the line of intestinal sutures. Good results were obtained.


Potassium bromate-induced oxidative stress, genotoxicity and cytotoxicity in the blood and liver cells of mice.
Potassium bromate (KBrO3) is an oxidising agent that is extensively used as a food additive, it is also a product of cosmetic and pharmaceutical relevance. The objective of this study was to evaluate the oxidative stress, genotoxicity, and apoptosis induced by KBrO3 in an experimental animal model. To study the toxic effects and oxidative stress, different doses of KBrO3 below LD50 (The half maximal lethal dose, 50, 100 and 150 mg/kg body weight) were given intraperitoneally to the mice for multiple time periods (24, 48, and 72 h). The results showed that KBrO3 significantly induces oxidative damage by increasing the levels of reactive oxygen species (ROS) and lipid peroxidase and depleted the levels of catalase (CAT), superoxide dismutase (SOD) and glutathione (GSH) enzymes in the serum and liver. Moreover, a significant increase of chromosomal aberrations in bone marrow cells and an elevated incidence of micronuclei in the peripheral blood of mice were observed. KBrO3 induces 3 ´ -OH end double-strand DNA breaks, which was evident in liver sections of the treated mice, and increases the percentage of apoptotic cells, as observed in TUNEL assays and flow cytometry analysis. The present findings indicate that KBrO3 induces oxidative stress, genotoxicity, and cytotoxicity in a dose- and time-dependent manner in mice.


Profile of Acute Kidney Injury in Hospitalized Children with Idiopathic Nephrotic Syndrome.
To determine the incidence, risk factors and outcome of acute kidney injury (AKI) in hospitalized children with nephrotic syndrome. All consecutive hospitalized children (aged 1-14 years) with diagnosis of nephrotic syndrome between February 2016 and February 2017 were enrolled for the study. Children (aged 1-14 years) with features of nephritis, underlying secondary causes of nephrotic syndrome as well as children admitted for diagnostic renal biopsy and intravenous cyclophosphamide or rituximab infusion were excluded. A total of 73 children (81 admissions) were enrolled; incidence of AKI was 16% (95% CI, 9-23). On multivariate logistic regression analysis, furosemide infusion was observed as an independent risk factor for acute kidney injury (OR 23; 95% CI, 3-141; P<0.001). Out of 13 children with AKI, three died. Acute kidney injury in hospitalized children with nephrotic syndrome has high risk of mortality. Children receiving furosemide infusion should be closely monitored for occurrence of acute kidney injury.


A multiplex polymerase chain reaction to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus -species venerealis: use on UK isolates of C. fetus and other Campylobacter spp.
Subspeciation of Campylobacter fetus subsp. fetus (CFF) and Campylobacter fetus subsp. venerealis (CFV) is important for international animal import regulations. Phenotyping can be unreliable, and genotyping by techniques like pulsed field gel electrophoresis is difficult in routine diagnostic laboratories. A PCR subspeciation technique has been reported [Aust Vet J (1997) 75, 827]; we aimed to develop this PCR and investigate its use on UK C. fetus isolates. We augmented the PCR with further primers, and tested 76 isolates of C. fetus and 16 isolates of other Campylobacter spp. PCR failed to correlate well with phenotyping, especially for CFV. We characterized the amplicon of the CFV-specific primers (reported as plasmid derived, but unavailable on the public databases); and predicted a parA gene sequence, anticipated to be plasmid-associated. However, although plasmid isolations from selected CFV isolates demonstrated the presence of several plasmids, there was no correlation between plasmid profile and PCR result. Further, the parA sequence was not detected by PCR in any of the plasmid bands. This PCR is not suitable for subspeciation of C. fetus in the UK. The results suggest that this is a reflection of the presence of an unusual clone of CFV currently present in cattle in this country. PCR cannot substitute for phenotyping of C. fetus isolates in the UK. The reasons for failure of PCR genotyping may reflect local strains and/or plasmid profiles. Further study is required to better elucidate molecular sub-speciation of C. fetus.


Age at first episode of venereal syphilis in an aboriginal population: an application of survival analysis.
To identify the age at which screening for syphilis could be ceased in Kimberley Aboriginal people who had remained seronegative and whether this age was the same for people with and without a past history of gonorrhea. Retrospective cohort study. The Kimberley region in the far north of Western Australia. Kimberley Aboriginal residents. Age at seroconversion. Survival analysis showed that seroconversion due to venereal syphilis occurred largely during the years between the late teens and early 20s. The probability of seroconverting by a given age was related to the time at which the subject's last gonorrhea infection occurred. As a result of this study, annual periodic screening for syphilis is now offered to Kimberley Aboriginal people only between the ages of 15 and 25 years, allowing for the redistribution of resources to other activities that have greater potential to decrease the incidence of all STDs.


A survey of opinions on emergency contraception in young women in Southern Italy.
To assess opinions and attitudes towards the use of hormonal emergency contraception (HEC) in a sample of young women in Puglia, Italy. A self-administered standardised anonymous questionnaire was administered to female school-leavers who were attending a preparatory course at the University of Bari. Seven hundred and fifty-four women, with an average age of 19.5 years, completed the questionnaire. Sixteen percent of respondents had used HEC and 3% had used it more than once. The reasons given for resorting to HEC were: rupture of the condom (57%), the lack of thought about the risk of an undesired pregnancy (25%), and the unavailability of contraceptives at the time of coitus (18%). The opinions expressed by the interviewees in our survey do not suggest that the use of HEC would increase the risks linked to sexual behaviour. In Italy, over-the-counter availability of HEC should be implemented in parallel with the improvement of sex education programmes aimed at the young.


Effects of plasticizers and plastic bags on granulocyte function during storage.
The influence of the plasticizers, di-(2-ethylhexyl)phthalate (DEHP) and tri-(2-ethylhexyl)trimellitate (TOTM), on granulocyte function was examined. Polyvinyl chloride (PVC) bags with DEHP (DEHP-PVC) leaked DEHP into plasma, but TOTM did not dissolve in plasma under the same conditions. Glow discharge treatment inhibited the leakage of DEHP from DEHP-PVC bags. Depending on the amount of DEHP added into granulocyte suspension, chemotaxis and bactericidal activity decreased, but cell counts and phagocytosis were not affected. During storage for 24 h at 22 degrees C, granulocyte function decreased greatly in DEHP-PVC, but was well maintained in the bags which did not leak plasticizers, TOTM-PVC and glow-discharged DEHP-PVC.


Molecular cloning of large alternative transcripts based on comparative phylogenetic analysis and exploration of an EST database.
In animals, a gene may be 50 kb or over and contain multiple alternative transcripts with sequences that are not experimentally validated. Under these special circumstances, PCR-based cloning may become very difficult. Here a simple cloning strategy is described using the mNLRC5 gene as an example. We performed comparative phylogenetic analysis between murine and human NLR protein families to anchor the translation start codon, searched an EST database with the 3' end of the genomic DNA sequence to obtain ESTs from the farthest 3' end of the gene, and isolated the full-length CDS of the mNLRC5 of about 6 kb through conventional RT-PCR and 3' RACE.


The mediation and moderation effect of social support on the relationship between opioid misuse and suicide attempts among native American youth in New Mexico: 2009-2019 Youth Risk Resiliency Survey (NM-YRRS).
Suicide attempt and opioid misuse continue to be major behavioral health challenges among American Indians and Alaska Natives (AI/AN). The aim of the study is to evaluate the mediating and moderating role that social support (SS) plays in their association among AI/AN high-school students in New Mexico (NM). An aggregated NM Youth Resiliency and Risk Survey (NM-YRRS, 2009-2019: odd years) dataset was used. Multivariable logistic regression modeling and mediation analysis were conducted while adjusting for confounding variables. Overall, 12.0 and 14.0% of AI/AN students reported opioid misuse and suicide attempt, respectively. The adjusted odds ratio of suicide attempt in students with high SS relative to low SS who misused opioids was 0.43 (p-value = 0.007). The effect of high SS relative to low SS among males who misused opioids was more pronounced (AOR = 0.24, p-value < 0.0001) compared to females (AOR = 0.43, p-value = 0.007). Relative to low SS, high SS was protective for suicide attempt among AI/AN students who misused opioids and attended school in off-reservation (AOR = 0.42, p-value = 0.012) communities, rural communities (AOR = 0.44, p = 0.040), and in communities that are both rural and off-reservation (AOR = 0.39, p = 0.035). Overall, 23.64, and 41.05% of the association between opioid misuse, and suicide attempt was mediated and moderated by SS, respectively. The mediation effect of SS was lowest for rural, on-reservation schools. More resources need to be allocated to rural on-reservation schools to enhance social support. The study highlights key insights into the significant role SS plays in promoting health and mitigating the association between opioid misuse and suicide attempt.


Postoperative Lactate Levels and Hospital Length of Stay After Cardiac Surgery.
The objective of this study was to characterize the association between lactate levels and hospital length of stay (LOS) after cardiac surgery. A retrospective study using prospectively collected data from the Society of Thoracic Surgeons adult cardiac surgery database. A tertiary-care hospital. Patients in the database who presented for major cardiac surgery between 2002 and 2014 and whose lactate level was measured within 3 hours after skin closure. None. The authors performed multivariable linear regression with adjustment for more than 30 variables to assess the association between postoperative lactate levels and hospital LOS. The study included 1,208 patients whose median LOS was 6 days (quartiles: 5, 9). Median LOS in the low-, moderate-, and high-lactate groups was 5 days (quartiles: 4, 7), 6 days (quartiles: 5, 9) and 9 days (quartiles: 6, 17), respectively; p<0.001. In multivariable analysis, patients with a moderate lactate level had a 1.08 times (95% CI: 1.00-1.17; p = 0.04) longer LOS compared with those with a low lactate level. Patients with a high lactate level had a 1.12 times (95% CI: 1.00-1.26; p = 0.04) longer LOS compared with those with a low lactate level. Lactate levels also were associated with intensive care unit LOS and nonsurgical postoperative complications. Postoperative lactate levels are associated with increased hospital LOS for patients undergoing major cardiac surgery.


Electrochemistry of purine derivatives. 1: Direct determination for the antiviral drug 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine by anodic differential pulse voltammetry.
Differential pulse voltammetry at a stationary glassy carbon electrode was used for the sensitive and selective analysis of a potent new antiviral analogue of 2-deoxyguanosine in a pharmaceutical formulation. In the electrochemical method for analysis of 9-[1,(3-dihydroxy-2-propoxy)methyl] guanine (1), an electroactive internal standard (uric acid) was used. Linear peak current-concentration relationships were obtained at 1 concentrations of 0.4-2.0 mM, with a quantitation limit of 0.1 mM. Degraded solutions of 1 were assayed directly by differential pulse voltammetry and also by two chromatographic methods to demonstrate the specificity of the electrochemical method. The voltammetric method reliably provides accurate and reproducible results in considerably less time than conventional chromatographic analysis.


Unique synthesis of hollow Co3O4 nanoparticles embedded in thin Al2O3 nanosheets for enhanced lithium storage.
The designed synthesis of advanced nanocomposite architecture is significant for its applications in energy storage, catalysis, sensing, etc. Herein, thin Al2O3 hexagonal nanosheets with encapsulated hollow Co3O4 nanoparticles (Co3O4-HNPs) are successfully synthesized by using Co6Al2CO3(OH)16·4H2O nanosheets as templates followed by a two-step annealing process. When used as an anode material in lithium ion batteries (LIBs), the homogeneous Co3O4-HNP/Al2O3 nanosheet composite exhibits an excellent performance with high reversible capacity and rate capability, and enhanced cycling stability.


Effect of rofecoxib on the pharmacokinetics of chronically administered oral contraceptives in healthy female volunteers.
The effect of rofecoxib, a highly selective cyclooxygenase (COX)-2 inhibitor, on the pharmacokinetics of ethinyl estradiol (EE) and norethindrone (NET), two common components of a combination oral contraceptive product, was examined. A double-blind, two-period crossover study was conducted in 18 healthy women who received ORTHO-NOVUM 1/35, a combination of EE (35 microg) and NET (1 mg), concurrently for 14 days with either 175 mg rofecoxib or matching placebo during two consecutive menstrual cycles. Plasma was sampled for EE, NET, sex hormone binding globulin (SHBG), and albumin. The AUC(0-24 h) geometric mean ratio (GMR: rofecoxib/placebo) with corresponding 90% confidence interval (CI) of EE and NET was 1.13 (1.06, 1.19) and 1.18 (1.13, 1.24), respectively. The Cmax GMR of EE and NET was 1.06 (0.98, 1.16) and 1.04 (0.99, 1.09), respectively. In each case, the 90% CIs satisfied the predefined bioequivalence limits of (0.80, 1.25). Measures of SHBG and albumin and routine clinical and laboratory safety parameters showed no clinically meaningful changes. The addition of rofecoxib to the oral contraceptive was not associated with any clinically important changes in EE or NET pharmacokinetics and thus would not be anticipated to influence the efficacy of this contraceptive regimen.


Influence of postharvest ultrasounds treatments on tomato (Solanum lycopersicum, cv. Zinac) quality and microbial load during storage.
Whole tomato fruits were treated at ultrasonic power levels from 10% to 100%, and at a constant frequency of 45 kHz, for different times (1-19 min). A central composite rotatable design (CCRD) was applied to optimise ultrasonic treatments for tomato quality (colour, texture and total phenolic content (TPC)) maintenance. According to response surface analysis, the optimal treatment parameters were 55%_10 min, 80%_15 min and 100%_19 min. At these conditions, and especially at higher power levels, a maximum retention of colour and texture, as well as an increase of TPC and microbial reduction were obtained in comparison with untreated fruits during 15 storage days at 10°C. The ultrasounds treatment was found to be effective in delaying colour development and texture losses, preserving sensorial quality of whole tomato, with increase of TPC and microbial load reduction. Moreover, this postharvest treatment can be used as an alternative for extending fresh fruits shelf-life.


A direct and melanopsin-dependent fetal light response regulates mouse eye development.
Vascular patterning is critical for organ function. In the eye, there is simultaneous regression of embryonic hyaloid vasculature (important to clear the optical path) and formation of the retinal vasculature (important for the high metabolic demands of retinal neurons). These events occur postnatally in the mouse. Here we have identified a light-response pathway that regulates both processes. We show that when mice are mutated in the gene (Opn4) for the atypical opsin melanopsin, or are dark-reared from late gestation, the hyaloid vessels are persistent at 8 days post-partum and the retinal vasculature overgrows. We provide evidence that these vascular anomalies are explained by a light-response pathway that suppresses retinal neuron number, limits hypoxia and, as a consequence, holds local expression of vascular endothelial growth factor (VEGFA) in check. We also show that the light response for this pathway occurs in late gestation at about embryonic day 16 and requires the photopigment in the fetus and not the mother. Measurements show that visceral cavity photon flux is probably sufficient to activate melanopsin-expressing retinal ganglion cells in the mouse fetus. These data thus show that light--the stimulus for function of the mature eye--is also critical in preparing the eye for vision by regulating retinal neuron number and initiating a series of events that ultimately pattern the ocular blood vessels.


Measuring the relationship between employees' health risk factors and corporate pharmaceutical expenditures.
This study demonstrates the relationship between self-reported health risk factors on a health risk appraisal and pharmaceutical expenditures for a large employer. A total of 3554 employees who were participants in a pharmacy benefit plan for the entire year of 2000 completed a health risk appraisal. As the number of self-reported health risk factors increased from zero to six or more, corporate pharmaceutical costs increased in a stepwise manner: US dollars 345, 443, 526, 567, 750, 754, and 1121 US dollars, respectively. After controlling for age, gender, and the number of self-reported diseases, each additional risk factor was associated with an average annual increase in pharmacy claims costs of 76 US dollars per employee. Specific health risks were associated with significantly higher expenditures. The results provide estimates of incremental expenditures associated with common, potentially modifiable risk factors. Pharmaceutical expenditures should be considered by corporations in their estimates of total health-related costs and in prioritizing disease management initiatives based on health risk appraisal data.


Discovering small-molecule therapeutics against SARS-CoV-2.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly become a global health pandemic. The lack of effective treatments, coupled with its etiology, has resulted in more than 400,000 deaths at the time of writing. The SARS-CoV-2 genome is highly homologous to that of SARS-CoV, the causative agent behind the 2003 SARS outbreak. Based on prior reports, clinicians have pursued the off-label use of several antiviral drugs, while the scientific community has responded by seeking agents against traditional targets, especially viral proteases. However, several avenues remain unexplored, including disrupting E and M protein oligomerization, outcompeting host glycan-virus interactions, interfering with the heparan sulfate proteoglycans-virus interaction, and others. In this review, we highlight some of these opportunities while summarizing the drugs currently in use against coronavirus 2019 (COVID-19).


Sensitivity, Specificity and Accuracy of Ultrasound in Diagnosis of Breast Cancer Metastasis to the Axillary Lymph Nodes in Chinese Patients.
The use of ultrasound in the diagnosis of axillary lymph node metastases from breast cancer in a Chinese population was investigated. Data for 1,049 with breast cancer were retrospectively collected. All patients had undergone pre-operative axillary ultrasound and then axillary lymph node dissection. The sensitivity, specificity and accuracy of axillary ultrasound in this cohort were 69.4%, 81.8% and 77.0%, respectively. The overall false-negative rate of ultrasound images was 30.6% (123/402). False-negative ultrasound rates for pathologic N1, N2 and N3 patients were 46.2%, 21.8% and 9.3%, respectively. In patients with stage T1 disease and fewer than three metastatic lymph nodes, the false-negative ultrasound rate was 52.2% (47/90). Moreover, breast cancer patients with a false-negative axillary ultrasound were more likely to have a large tumor (p < 0.001) and high tumor grade (p = 0.009). However, there were no statistically significant differences between accuracy of axillary ultrasound and age of patients or experiences of ultrasound practitioners. In conclusion, the sensitivity, specificity and accuracy of ultrasound in the diagnosis of breast cancer metastasis to the axillary lymph nodes in Chinese patients were assessed. These data could help us to carefully use axillary ultrasound to diagnose and predict breast cancer axillary lymph node metastasis.


Validity of longitudinal sections for determining the apical constriction.
To validate the use of longitudinal sections against cross sections using micro-CT for disclosing the topography and location of the apical constriction. Seventy extracted human teeth with 117 completely developed roots were micro-CT scanned and reconstructed at a voxel size of 27 μm. The 3DSlicer program was used to navigate the longitudinal sections parallel to the long axis of the canal and also to rotate and tilt the views. Each root canal was evaluated in both mesio-distal and bucco-lingual planes. Constriction topographies were identified as described in the literature. In each canal, the number of different topographies detected was recorded. Further, serial cross-sectional analysis of the apical portion of the canal was performed. Reconstructed plots of canal areas were assessed to locate the constriction and determine its form. A descriptive analysis of both longitudinal and cross section methods was conducted. In each canal, the frequency of constriction forms was calculated in the mesio-distal or bucco-lingual aspects and the 99% confidence interval was computed. When both aspects of the longitudinal sections were pooled, all root canals had two or more topographies and consequently different locations of the apical constriction. In contrast, cross-sectional analysis constantly yielded one constriction form per canal. Compared to cross-sectional analysis, longitudinal sections of the root canal conveyed inconsistent results regarding the topography and the location of the apical constriction.


Dietary trends of 10- and 13-year-old children in a biracial community--the Bogalusa Heart Study.
Dietary intakes of two cross-sectional cohorts of 10-year-old children were collected in 1973 and 1976 to examine dietary trends over time. A longitudinal sample of the initial 1973 cohort examined at both 10 and 13 years of age identified changes in patterns and aging effect on intake. Significantly higher energy, total protein, and polyunsaturated fatty acids were noted for both 10- and 13-year-olds in 1976 compared with the 1973 sample. Saturated fat intake per 1,000/kcal was significantly lower but polyunsaturated fat was significantly higher per 1,000/kcal for the second 10-year-old group. Boys consumed more energy, protein, saturated fat, cholesterol, and iron than did girls. A sex X age interaction was noted for energy, sucrose, starch, and potassium, with boys ingesting more at 13 than at 10 years. A race X age interaction occurred for sodium, with black children having higher levels than white children at 10 but not at 13 years. Statistically significant correlations were noted between intake at 10 and 13 years of age for protein, cholesterol, and potassium for boys. Eighty-five percent of all children exceeded the protein Recommended Dietary Allowance. Over half the children exceeded the maximum recommended sodium range; potassium intakes were low. Excessive intakes of saturated fat, sucrose, animal protein, and sodium with low potassium and vegetable protein intakes create a potential health risk, influencing cardiovascular risk.


The role of primary plastic surgery in the management of open fractures.
A study was undertaken to determine the requirement for primary plastic surgery in the treatment of open fractures. We reviewed 3297 consecutive open fractures in a 22-year period in a defined population. Analysis showed that 12.6% of patients required primary plastic surgery with 5.6% being treated with split skin grafting and 7.2% with a flap. Only 3.5% of open upper limb fractures required primary plastic surgery compared to 27.9% of open lower limb fractures. The fractures that required most primary plastic surgery were those of the femoral diaphysis and all fractures between the proximal tibia and the midfoot. The incidence of open fractures that require primary plastic surgery was 28/10(6)/year. The incidence in open upper and lower limb fractures was 5.3/10(6)/year and 22.7/10(6)/year respectively. Using these figures it is possible to estimate the numbers of open fractures that will require primary plastic surgery each year in the United Kingdom.


Dual β-oxidation pathway and transcription factor engineering for methyl ketones production in Saccharomyces cerevisiae.
Methyl ketones (MK) are highly valuable fatty acid derivatives with broad applications. Microbes based biosynthesis represents an alternative route for production of these usually fossil based chemicals. In this study, we reported metabolic engineering of Saccharomyces cerevisiae to produce MK, including 2-nonanone, 2-undecanone, 2-tridecanone and 2-pentadecanone. Besides enhancing inherent peroxisomal fatty acids β-oxidation cycle, a novel heterologous cytosolic fatty acids β-oxidation pathway was constructed, and this resulted in an increased production of MK by 2-fold. To increase carbon fluxes to methyl ketones, the supply of precursors was enhanced by engineering lipid metabolism, including improving the intracellular biosynthesis of acyl-CoAs, weakening the consumption of acyl-CoAs for lipids storage, and reinforcing activation of free fatty acids to acyl-CoAs. Hereby the titer of MK was improved by 7-fold, reaching 143.72 mg/L. Finally, transcription factor engineering was employed to increase the biosynthesis of methyl ketones and it was found that overexpression of ADR1 can mimic the oleate activated biogenesis and proliferation of peroxisomes, which resulted in a further increased production of MK by 28%. With these modifications and optimization, up to 845 mg/L total MK were produced from glucose in fed-batch fermentation, which is the highest titer of methyl ketones reported produced by fungi.


Activation of protein kinase C inhibits kainate-induced currents in oocytes expressing glutamate receptor subunits.
The effect of protein kinase C (PKC) activation on maximal kainate (KA)-induced currents was studied in Xenopus oocytes expressing the glutamate receptor (GluR) subunits GluR3, GluR1 + 3, GluR2 + 3, and GluR6. The PKC activator phorbol 12-myristate 13-acetate (PMA) inhibited peak KA responses in a time-dependent manner. The magnitude of inhibition was greatest in GluR6-expressing oocytes. Desensitizing KA currents characterized by a peak, transient current followed by a slower, desensitizing current were observed in oocytes expressing GluR3 and GluR1 + 3 receptors. PMA inhibited the desensitization, and this effect could be observed before PMA's inhibition of peak current amplitude. PMA-mediated inhibition of both desensitization and peak current amplitude was prevented by intracellular injection of the protein kinase C (PKC) inhibitor peptide. These results suggest that the function of GluRs is regulated by PKC-dependent phosphorylation.


Esmolol attenuates lung injury and inflammation in severe acute pancreatitis rats.
Recent studies suggest that beta-adrenergic blockers attenuate systemic inflammation and improve survival in sepsis. We investigated whether esmolol can reduce lung injury and modulate inflammatory response in a rat model of severe acute pancreatitis (SAP). A taurocholate-induced SAP was used, with or without continuously intravenous pumping of esmolol (15 mg/kg/h). Heart rate and arterial pressure were monitored. Nine hrs after esmolol administration, blood was drawn for blood gas analyses and cytokine (interleukin(IL)-6, tumor necrosis factor (TNF)-α) detections, lungs and pancreata were isolated for measurements of myeloperoxidase (MPO) activity and histological damage. In an additional 20 animals, rats were randomized into SAP or SAP + esmolol groups to assess effects of esmolol on survival time. Treatment with esmolol was associated with improved survival time (11.1 ± 1.6 h vs. 9.2 ± 2.0 h, p = 0.044) and less severe disease, as assessed by lung and pancreas histology. Blood gas analyses were ameliorated in esmolol group. Arterial PO2 increased (109.7 ± 12.4 mmHg vs 93.9 ± 4.1 mmHg, p = 0.008) while lactate levels (2.1 ± 0.5 vs 3.1 ± 0.7 mmol/L, p = 0.001) decreased in SAP + esmolol group as compared with SAP group. Esmolol treatment also abated the increase in bronchoalveolar lavage fluid protein and proinflammatory cytokines. Furthermore, esmolol reduced SAP-induced plasma amylase activity (p = 0.02), blunted the expression of TNF-α (p = 0.003) and IL-6 (p < 0.001), and decreased pancreas/lung MPO activities. Continuous infusion of esmolol, a selective beta-1 adrenergic blocker, improves outcome, reduces inflammatory responses and also offers lung and pancreas protective effects in SAP rats. This may offer novel therapeutic strategies in treating patients suffering from SAP.


The effect of an acute phase response on tissue carotenoid levels of growing chickens (Gallus gallus domesticus).
Plasma, liver and skin carotenoids decrease following infectious disease challenges. Since these challenges often involve substantial host pathology and chronic immune responses, the mechanism underlying altered carotenoid deposition is unclear. Therefore, changes in tissue carotenoid levels were examined during an acute phase response induced by lipopolysaccharide (LPS) or interleukin-1 (IL-1). In two experiments, chicks were hatched from carotenoid-deplete eggs (n=28, n=64, respectively) and fed 0, 8 or 38 mg carotenoids (lutein+canthaxanthin)/kg diet. For chicks fed 38 mg carotenoids, but not those fed 0 or 8 mg, LPS generally reduced plasma lutein, canthaxanthin and total carotenoids (P<0.05), and liver lutein, zeaxanthin, canthaxanthin and total carotenoids (P<0.05). Additionally, LPS reduced thymic total carotenoids (P=0.05) and increased thymocyte lutein (P=0.07), zeaxanthin (P=0.07) and total carotenoids (P=0.07). Finally, LPS increased bursal canthaxanthin (P<0.01), but had no effect on shank carotenoids (P>0.5). In chicks hatched from carotenoid-replete eggs (n=36) and fed dietary lutein (38 mg/kg diet), LPS reduced plasma and liver zeaxanthin and liver total carotenoids (P<0.05); IL-1 reduced plasma and liver lutein, zeaxanthin and total carotenoids (P<0.05). Therefore, an acute phase response plays a role in reduced tissue carotenoids during infectious disease.


Bed nucleus of the stria terminalis NMDA receptors and nitric oxide modulate contextual fear conditioning in rats.
The bed nucleus of the stria terminalis (BNST) modulates anxiety-like responses, including conditioned emotional responses. Evidence suggests that glutamatergic neurotransmission in the BNST plays a role in the modulation of defensive responses. However, little is known about the involvement of glutamate NMDA receptor activation within the BNST, and its resultant increase in nitric oxide (NO) levels, in the expression of contextual fear conditioning (CFC). We investigated whether the antagonism of NMDA receptors or the reduction of NO levels in the BNST would attenuate behavioral and autonomic responses (i.e. increase in arterial pressure and heart rate, and decrease in tail cutaneous temperature) of rats submitted to a CFC paradigm. Intra-BNST infusion of AP7, an NMDA receptor antagonist, attenuated both behavioral and autonomic changes induced by CFC. Similar results were observed with NPLA and c-PTIO, an nNOS inhibitor and an NO scavenger, respectively. A positive correlation between BNST NO levels and the time spent in freezing behavior was also observed for animals submitted to the CFC. These findings indicate that the expression of CFC involves a facilitation of BNST NMDA receptor-NO signaling. This article is part of the Special Issue entitled 'Ionotropic glutamate receptors'.


Irpex lacteus polysaccharide exhibits therapeutic potential for ovarian fibrosis in PCOS rats via the TGF-β1/smad pathway.
Polycystic ovarian syndrome (PCOS) is one of the commonest endocrinopathies in childbearing women. The research was conducted to assess the impact of Irpex lacteus polysaccharide (ILP, 1000 mg/kg) on the letrozole (1 mg/kg)-induced PCOS model in female rats. Metformin (Met, 265 mg/kg) as the positive control. The study suggested that ILP restored the estrous cycle in rats with PCOS as well as lowered relative ovarian weight and body weight, in comparison to normal. Rats with PCOS showed improvement in ovarian structure and fibrosis when given ILP. ILP decreased the testosterone (T), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), total cholesterol (TC), luteinizing hormone (LH), homeostasis model assessment-insulin resistance (HOMA-IR), fasting blood glucose (FBG), and insulin (INS) levels and elevated the follicle-stimulating hormone (FSH) and estrogen (E2) levels in PCOS rats. In addition, ILP increased the content of superoxide dismutase (SOD) in serum and the antioxidant enzymes (Prdx3, Sod1, Gsr, Gsta4, Mgst1, Gpx3, Sod2 and Cat) expression levels in the ovaries and decreased the serum expression of malondialdehyde (MDA). In addition, ILP treatment slowed down the process of the fibrosis-associated TGF-β1/Smad pathway and downregulated α-smooth muscle actin (α-SMA) and connective tissue growth factor (CTGF) levels in PCOS rats ovaries. According to these findings, ILP may be able to treat letrozole-induced PCOS in rats by ameliorating metabolic disturbances, sex hormone levels, oxidative stress, and ovarian fibrosis.


Epidemiological, virological, and entomological characteristics of dengue from 1978 to 2009 in Guangzhou, China.
To understand its unprecedented resurgence, we examined the epidemiological, virological, and entomological features of dengue in Guangzhou during 1978-2009. Cases reported to the Guangzhou Centre for Disease Control and Prevention and data from virological and entomological surveillance were analyzed from three periods: 1978-1988, 1989-1999, and 2000-2009. Although cases decreased over time: 6,649 (1978-1988) to 6,479 (1989-1999) to 2,526 (2000-2009), geographical expansion resulted in districts with an average incidence >2.5/100,000, increasing from five (1978-1988, 1989-1999) to seven (2000-2009). Age distribution (mean age: 34.9 years) provided a trend of increasing dengue incidence among adults, and there was a significantly higher incidence among men with a sex ratio of 1.15:1 (P<0.001). Cases occurred from May through November with a peak between August and October, and a long-term trend was characterized by a three to five-year cyclical pattern. The most frequently isolated serotypes were DENV-2 (1978-1988) and DENV-1 (1989-1999 and 2000-2009). Seasonal fluctuations in immature densities of Aedes albopictus (sole transmission vector in Guangzhou) were consistent with the dengue seasonality. After a 30-year apparent absence, DENV-3 had reemerged in 2009. The current epidemiological situation is highly conducive to periodic dengue resurgences. Thus, a high degree of surveillance and strict control measures in source reduction should be maintained.


Heat shock proteins and the calcineurin-crz1 signaling regulate stress responses in fungi.
The heat shock proteins (Hsps) act as a molecular chaperone to stabilize client proteins involved in various cell functions in fungi. Hsps are classified into different families such as HSP90, HSP70, HSP60, HSP40, and small HSPs (sHsps). Hsp90, a well-studied member of the Hsp family proteins, plays a role in growth, cell survival, and pathogenicity in fungi. Hsp70 and sHsps are involved in the development, tolerance to stress conditions, and drug resistance in fungi. Hsp60 is a mitochondrial chaperone, and Hsp40 regulates fungal ATPase machinery. In this review, we describe the cell functions, regulation, and the molecular link of the Hsps with the calcineurin-crz1 calcium signaling pathway for their role in cell survival, growth, virulence, and drug resistance in fungi and related organisms.


Orthodeoxia Platypnea Syndrome in the Setting of Cholangiocarcinoma: A Case Report.
Platypnea-orthodeoxia syndrome is a rare cause of positional hypoxemia and dyspnea. We present the case of a 54-year-old man with right-to-left shunting through a patent foramen ovale in the setting of metastatic cholangiocarcinoma resulting in platypnea-orthodeoxia syndrome. The shunt was originally not visualized on cardiac magnetic resonance imaging but later detected with transesophageal echocardiography. This case highlights the importance of complimentary multimodality cardiac imaging in the diagnosis of both common and uncommon disorders.


FeCl3-Assisted Niobium-Catalyzed Cycloaddition of Nitriles and Alkynes: Synthesis of Alkyl- and Arylpyrimidines Based on Independent Functions of NbCl5 and FeCl3 Lewis Acids.
NbCl5-catalyzed [2 + 2 + 2] cycloaddition of nitriles with alkynes was used to synthesize pyrimidine derivatives. In this reaction, the use of individual Lewis acids, namely NbCl5 and FeCl3, is a key strategy for achieving the reaction using a catalytic amount of NbCl5. The roles of the two Lewis acids were investigated using FT-IR spectroscopy. The results showed that NbCl5 served as an efficient Lewis acid catalyst for nitrile activation, whereas FeCl3 showed stronger Lewis acidity toward pyrimidines, releasing NbCl5 into the catalytic cycle.


Constructing future scenarios as a tool to foster responsible research and innovation among future synthetic biologists.
The emerging field of synthetic biology, the (re-)designing and construction of biological parts, devices and systems for useful purposes, may simultaneously resolve some issues and raise others. In order to develop applications robustly and in the public interest, it is important to organize reflexive strategies of assessment and engagement in early stages of development. Against this backdrop, initiatives related to the concept of Responsible Research and Innovation (RRI) have also appeared. This paper describes such an initiative: the construction of future scenarios to explore the plausibility and desirability of potential synthetic biology innovations. We guided teams of synthetic biology students who participated in the large international Genetically Engineered Machines (iGEM) competition, in constructing scenarios aimed at exploring the plausibility and desirability of potential synthetic biology innovations. In this paper we aim to examine to what extent, and how, constructing such future scenarios contributes to RRI. In order to do so, we conducted observations and interviews to understand what kind of learning and reflection was promoted by constructing the scenarios in terms of four dimensions, which are discussed prominently in the literature on RRI: anticipation, inclusion, reflexivity and responsiveness. While we focus on how constructing future scenarios can contribute to strengthening RRI at a project (and individual) level, we also consider how far our experiment may foster RRI in the iGEM competition in general, and perhaps even inspire constructive collaboration between 'social scientists' and 'natural scientists' in the context of larger scientific research programmes.


Dissolved-Cl2 triggered redox reaction enables high-performance perovskite solar cells.
Constructing 2D/3D perovskite heterojunctions is effective for the surface passivation of perovskite solar cells (PSCs). However, previous reports that studying perovskite post-treatment only physically deposits 2D perovskite on the 3D perovskite, and the bulk 3D perovskite remains defective. Herein, we propose Cl2-dissolved chloroform as a multifunctional solvent for concurrently constructing 2D/3D perovskite heterojunction and inducing the secondary growth of the bulk grains. The mechanism of how Cl2 affects the performance of PSCs is clarified. Specifically, the dissolved Cl2 reacts with the 3D perovskite, leading to Cl/I ionic exchange and Ostwald ripening of the bulk grains. The generated Cl- further diffuses to passivate the bulk crystal and buried interface of PSCs. Hexylammonium bromide dissolved in the solvent reacts with the residual PbI2 to form 2D/3D heterojunctions on the surface. As a result, we achieved high-performance PSCs with a champion efficiency of 24.21% and substantially improved thermal, ambient, and operational stability.


Determination of stoichiometry and conformational changes in the first step of the P22 tail assembly.
Large oligomeric portal assemblies have a central role in the life-cycles of bacteriophages and herpesviruses. The stoichiometry of in vitro assembled portal proteins has been a subject of debate for several years. The intrinsic polymorphic oligomerization of ectopically expressed portal proteins makes it possible to form rings of diverse stoichiometry (e.g., 11-mer, 12-mer, 13-mer, etc.) in solution. In this study, we have investigated the stoichiometry of the in vitro-assembled portal protein of bacteriophage P22 and characterized its association with the tail factor gp4. Using native mass spectrometry, we show for the first time that the reconstituted portal protein (assembled in vitro using a modified purification and assembly protocol) is exclusively dodecameric. Under the conditions used here, 12 copies of tail factor gp4 bind to the portal ring, in a cooperative fashion, to form a 12:12 complex of 1.050 MDa. We applied tandem mass spectrometry to the complete assembly and found an unusual dimeric dissociation pattern of gp4, suggesting a dimeric sub-organization of gp4 when assembled with the portal ring. Furthermore, native and ion mobility mass spectrometry reveal a major conformational change in the portal upon binding of gp4. We propose that the gp4-induced conformational change in the portal ring initiates a cascade of events assisting in the stabilization of newly filled P22 particles, which marks the end of phage morphogenesis.


Production of disulfide-linked hirudin dimer by in vitro folding.
A simple process of in vitro folding has been developed for the preparation of hirudin dimer. A variant of recombinant hirudin with Asp33 replaced by Cys was expressed in yeast and isolated by HPLC. Crude Cys33-hirudin contains heterogeneous products that are made of one species of primary sequence. They were together reduced/denatured, and allowed to re-fold in the sodium bicarbonate buffer (pH 8.3) alone. Active, homogeneous Cys33-hirudin monomer folded spontaneously with a first order rate constant of 0.05 +/- 0.01 min-1, followed by the oxidation of two Cys33 to produce the pure dimer. The folding yield was 90%. On an equal weight basis, both Cys33-hirudin monomer and the dimer exhibit thrombin inhibitory activity comparable to that of wild-type hirudin. Due to the presence of an extra cysteine, the folding of active hirudin monomer (formation of three native disulfides) was accelerated by at least 12-fold.


Two novel mutations in the thyroglobulin gene as cause of congenital hypothyroidism: identification a cryptic donor splice site in the exon 19.
Thyroglobulin (TG) is a homodimeric glycoprotein synthesized by the thyroid gland. To date, 52 mutations of the TG gene have been identified in humans. The purpose of the present study was to identify and characterize new mutations in the TG gene. We report a French patient with congenital hypothyroidism, mild enlarged thyroid gland and low levels of serum TG. Sequencing of DNA, genotyping, expression of chimeric minigenes as well as bioinformatics analysis were performed. DNA sequencing identified the presence of compound heterozygous mutations in the TG gene: the paternal mutation consists of a c.3788-3789insT or c.3788dupT, whereas the maternal mutation consists of g.IVS19+3_+4delAT. Minigene analysis of the g.IVS19+3_+4delAT mutant showed that the exon 19 is skipped during pre-mRNA splicing or partially included by use of cryptic 5' splice site located to 100 nucleotides downstream of the wild type exon-intron junction. The c.3788-3789insT mutation results in a putative truncated protein of 1245 amino acids, whereas g.IVS19+3_4delAT mutation originates two putative truncated proteins of 1330 and 1349 amino acids. In conclusion, we show that the g.IVS19+3_+4delAT mutation promotes the activation of a cryptic donor splice site in the exon 19 of the TG gene. These results open up new perspectives in the knowledge of the mechanism of splicing for the TG pre-mRNA.


When Phosphosugars Meet Gold: Synthesis and Catalytic Activities of Phostones and Polyhydroxylated Phosphonite Au(I) Complexes.
The synthesis and characterization of P-chiral phosphonite-, phosphonate- and thiophosphonate-Au(I) complexes are reported. These novel ligands for Au(I) are based on glycomimetic phosphorus scaffolds, obtained from the chiral pool. The catalytic activities of these complexes are shown in the cyclization of allenols and the hydroamination of 2-(2-propynyl)aniline combined with an organocatalyzed reduction to the corresponding 2-phenyl tetrahydroquinoline. All described gold complexes present excellent catalytic activities.


Suppression of colon cancer growth by berberine mediated by the intestinal microbiota and the suppression of DNA methyltransferases (DNMTs).
The purpose of this study was to demonstrate the regulatory effect of berberine (BBR) on the intestinal microbiota and related epigenetics during the inhibition of colon cancer cell growth in vitro and in vivo. We used a nude mouse xenograft model with HT29 colon cancer cells to establish and divide into a model group and BBR group. The mice were treated for four weeks, and HT29 cells in the BBR group were cultured for 48 h. Cetuximab and the DNA transmethylase (DNMT) inhibitor 5-AZA-dC were added to HT29 cells. Tumour volume and weight were measured by hematoxylin-eosin (HE) staining for histopathological observation. Mouse faeces were collected, and the gut microbiota was analysed with 16S rDNA amplicons. The levels of cytokines in the supernatant of HT29 cells were measured by ELISA. A CCK-8 kit was used to examine the proliferation of HT29 cells, and RT‒PCR was used to measure the levels of c-Myc, DNMT1, DNMT3A, and DNMT3B. We found that BBR reduced the growth of colon cancer cells to a certain extent in vitro and in vivo, although the difference was not statistically significant compared with that in the model group. BBR significantly mediated the abundance, composition and metabolic functions of the intestinal microbial flora in mice with colon cancer. The effect of BBR on inflammatory cytokines, including IL-6, FGF, and PDGF, was not obvious, but BBR significantly downregulated IL-10 levels (P < 0.05) and reduced c-Myc, DNMT1, and DNMT3B levels (P < 0.05). Inhibiting DNMTs with 5-AZA-dC significantly suppressed the proliferation of HT29 cells, which was consistent with the effect of BBR. The inhibitory effect of berberine on colon cancer is related not only to the intestinal microbiota and its metabolic functions but also to the regulation of DNMTs.


Strategies for obstetricians and gynecologists to advance reproductive autonomy in a post-Roe landscape.
The monumental reversal of Roe vs Wade dramatically impacted the landscape of reproductive healthcare access in the United States. The decision most significantly affects communities that historically have been and continue to be marginalized by systemic racism, classism, and ableism within the medical system. To minimize the harm of restrictive policies that have proliferated since the Supreme Court overturned Roe, it is incumbent on obstetrician-gynecologists to modify practice patterns to meet the pressing reproductive health needs of their patients and communities. Change will require cross-discipline advocacy focused on advancing equity and supporting the framework of reproductive justice. Now, more than ever, obstetrician-gynecologists have a critical responsibility to implement new approaches to service delivery and education that will expand access to evidence-based, respectful, and person-centered family planning and early pregnancy care regardless of their practice location or subspecialty.


Prognostic Impact of EGFR Amplification and Visceral Pleural Invasion in Early Stage Pulmonary Squamous Cell Carcinomas Patients after Surgical Resection of Primary Tumor.
Over the last few decades, an increasing amount of information has been accumulated on biomarkers in non-small cell lung cancer (NSCLC). Despite these advances, most biomarkers have been identified in the adenocarcinoma histological subtype (AC). However, the application of molecular-targeted therapies in the prognosis and treatment of SCC in the clinical setting is very limited, becoming one of the main focus areas in research. Here, we prospectively analyzed the frequency of numerical/structural abnormalities of chromosomes 5, 7, 8, 9, 13 and 22 with FISH in 48 pulmonary SCC patients. From a total of 12 probes, only abnormalities of the 7p12 and 22q12 chromosomal regions were identified as unique genetic variables associated with the prognosis of the disease. The study for these two chromosomal regions was extended to 108 patients with SCC. Overall, chromosome losses were observed more frequently than chromosome gains, i.e., 61% versus 19% of all the chromosome abnormalities detected. The highest levels of genetic amplification were detected for the 5p15.2, 7p12, 8q24 and 22q11 chromosome bands, of which several genes are potentially involved in the pathogenesis of SCC, among others, include the EGFR gene at chromosome 7p12. Patients who displayed EGFR amplification (n = 13; 12%) were mostly older than 65 years (p = 0.07) and exclusively patients in early T-primary tumor stage (pT1−pT2; p = 0.03) with a significantly shortened overall survival (OS) (p ≤ 0.001). Regarding prognosis, the clinical, biological, and histopathologic characteristics of the disease that displayed a significant adverse influence on OS in the univariate analysis included patients older than 65 years (p = 0.02), the presence of lymph node involvement (p = 0.005), metastasis (p = 0.01) and, visceral pleural invasion (VPI) at diagnosis (p = 0.04). EGFR amplification also conferred an adverse impact on patient OS in the whole series (p = 0.02) and especially in patients in early stages (pT1−pT2; p = 0.01). A multivariate analysis of the prognostic factors for OS showed that the most informative combination of independent variables to predict an adverse outcome was the presence of VPI and/or EGFR amplification (p < 0.001). Based on these two variables, a scoring system was built to stratify patients into low- (no adverse features: score 0; n = 69), intermediate- (one adverse feature: score 1; n = 29) and high-risk (two adverse features: score 2; n = 5) groups, with significantly different (p = 0.001) OS rates at 50 months, which were as following: 32%, 28% and 0%, respectively. In the present study, we show that the presence of a high level of 7p12 (EGFR) amplification, exclusively detected in early stage SCC (pT1−pT2), is an independent adverse prognostic factor for OS. The identification of the EGFR gene copy number using FISH techniques may provide a more accurate diagnosis of high-risk populations after the complete resection of the primary tumor. When combined with VPI, three groups of pulmonary SCC were clearly identified that show the extent of the disease. This is of such importance that further prospective studies are necessary in larger series of SCC patients to be classified at the time of diagnosis. This could be achieved with the combined assessment of 7p12 amplification and VPI in primary tumor samples.


Description of the new species Coptera tonic (Hymenoptera, Diapriidae), a pupal parasitoid of Rhagoletis juniperina Marcovitch (Diptera, Tephritidae), and revised partial keys to Nearctic Coptera Say.
A new species of the parasitic wasp Coptera Say was previously distinguished from other species via correspondence between ecological (host) differences and DNA barcodes. A description and figures for Coptera tonic sp. nov., along with revisions to existing keys that allow it to be distinguished from other Nearctic species without the aid of molecular characters, is provided in this work.


Gastric outlet obstruction due to a pancreatic pseudocyst: report of two cases.
Two cases of gastric outlet obstruction, secondary to a pseudocyst of the pancreas, are described. The striking radiologic pictures are shown. The pathogenesis of this complication is discussed and a greater awareness of this entity is stressed.


The accuracy and validity of the Sheffield Features of Gait Tool.
Gait is now widely used in the UK as a contributor to identification, and increasing interest is being shown in its use in both Europe and the US. One of the long standing criticisms of the use of gait as evidence has been the lack of a validated standard methodology. With the publication of the 'Code of practice for forensic gait analysis', and the adoption of the code as part of the 'Codes of Practice and Conduct for forensic science providers and practitioners in the Criminal Justice System' by the Forensic Science Regulator, forensic gait analysts are now required to provide evidence of the testing of the methods used. The Sheffield Features of Gait Tool is specifically designed to assist observational gait analysis in the forensic context, and was developed by forensic gait analysis practitioners based on their casework and trial experience. Birch et al 2019 reported the findings of a study undertaken to assess the repeatability and reproducibility of the tool. This paper reports the findings of a study undertaken to assess the accuracy with which analysts identified features of gait when using the tool. Fourteen participants, with experience in observational gait analysis, viewed footage of computer generated avatars walking, and completed the features of gait tool on multiple occasions. The results showed a mean accuracy score of 134.92 out of a possible 180 (74.96%), a standard deviation of 9.49 (5.27%) and a coefficient of variation of 7.03%, demonstrating a good degree of consistency between the scores (Cronbach's alpha <0.90; ANOVA p-value <0.05). The findings of this study, coupled with those of the Birch et al 2019 study which showed there to be good levels of both repeatability and reproducibility of observations of features of gait made by the participants, suggest that the Sheffield Features of Gait Tool is a valid and fit for purpose method of observing and recording features of gait in the forensic context. The use of the tool provides the basis of a standardised methodology for observational gait analysis in the forensic context.


Biochemical characterization of Escherichia coli DNA helicase I.
The gene product of F tral is a bifunctional protein which nicks and unwinds the F plasmid during conjugal DNA transfer. Further biochemical characterization of the Tral protein reveals that it has a second, much lower, Km for ATP hydrolysis, in addition to that previously identified. Measurement of the single-stranded DNA-stimulated ATPase rate indicates that there is co-operative interaction between the enzyme monomers for maximal activity. Furthermore, 18O-exchange experiments indicate that Tral protein hydrolyses ATP with, at most, a low-level reversal of the hydrolytic step during each turnover.


Assessment of T-cell receptor beta-chain diversity by heteroduplex analysis.
The aim of this work was to search for a simple and alternative approach to the currently used methodologies for the analysis of T-cell receptor repertoire diversity. To this end we studied whether the heteroduplex analysis could be adapted to study the clonality of the T-cell receptor beta chain (TCRBV). We therefore analyzed, by sequencing, the molecular characteristics of the V-D-J junctions of numerous TCRBV chains from a variety of patients and from normal individuals, and compared the results with those obtained with the heteroduplex analysis. The latter procedure involves the amplification of the target TCRBV chains and the denaturation and renaturation of the amplified product to permit the random association of the distinct DNA strands encoding the different junctional regions. Whereas amplified material from polyclonal lymphoid cells migrates on a polyacrylamide gel as a "smear" of bands composed of different-sized polyclonal PCR fragments, the mismatched chains derived from oligoclonal populations migrate as discrete "heteroduplexes" and can be separated from the matched "homoduplex" obtained from homogeneous clonal cells. Our results provide evidence demonstrating that heteroduplex analysis can successfully be applied to the analysis of T-cell clonality in a variety of samples and can be complementary or substitute for the standard approach of TCR cloning and multiple sequencing of junctional regions. Thus, the procedure should facilitate the implementation of the analysis of TCR in diagnostic routine and should find applications in numerous physiologic and pathologic conditions.


Expression of multiple alpha adrenergic receptor subtype messenger RNAs in the adult rat brain.
Multiple subtypes of alpha adrenergic receptors with CNS expression (alpha 1A, alpha 1B, alpha 2A and alpha 2C) have been identified through pharmacological and molecular biological means. To characterize the localization of these subtypes and attempt to correlate subtype expression with physiological significance, the expression of the mRNAs encoding the alpha 1A, alpha 1B, alpha 2A and alpha 2C adrenergic receptor subtypes was examined in the adult rat brain by in situ hybridization histochemistry. Each subtype demonstrated a unique pattern of distribution, with the alpha 1 adrenergic receptors more restricted in their distribution and the alpha 2 receptors more widespread. The alpha 1A was primarily localized in the olfactory bulb, intermediate layers of the cortex, the hippocampus and the reticular nucleus of the thalamus. The alpha 1B was expressed in intermediate and deep layers of the cortex, thalamus, hippocampus, dorsal raphe and cerebellum. Although the alpha 2A message was relatively low in abundance, it was identified in the olfactory bulb, cortex, hippocampus, locus coeruleus, pons and cerebellum. The alpha 2C messenger RNA was localized in the cortex (particularly cingulate), hippocampus, caudoputamen, pons and cerebellum. Multiple alpha adrenergic receptor subtypes have significant sequence homology and similar pharmacologic properties; however, they each possess a unique pattern of messenger RNA distribution throughout the brain. The multiplicity of subtypes of alpha adrenergic receptors in specific brain regions may dictate the physiological and pharmacological responses to catecholamines.


Tuftsin ameliorates splenic inflammatory injury by promoting neuropilin-1 in severe acute pancreatitis.
Severe acute pancreatitis (SAP)-associated spleen injury causing immune disturbances aggravates organs injuries, which contributes to higher mortality rate. However, there are no effective drugs to cure SAP-induced spleen injury. Here, we found that Tuftsin (TN) is effective for ameliorating SAP-induced pathological damage and inflammation of spleen, mainly via alleviating mitochondrial dysfunction, oxidative stress, ATP depletion and the expression of pro-inflammatory factors. We further found that TN promoted anti-inflammatory macrophage phenotype M2 via up-regulating NRP1 on macrophage in spleen during SAP. Meanwhile, EG00229 (an inhibitor of NRP1 bound to TN) weakened TN's therapeutic effect in SAP-associated spleen injury. And EG00229 also inhibited M2 macrophage, leading to increasing inflammasome formation. Additionally, EG00229 reduced the protective efficiency of TN on mitochondrial dysfunction, and inflammation injury via NRP1 in spleen caused by SAP. Similarly, siRNA-Nrp1 into macrophage also prevented TN's inhibition on apoptosis. These findings reveal that TN alleviates SAP-induced spleen injury by promoting NRP1.


[Agenesis of the left lung in combination with a persistent left superior vena cava and al cor triatriatum sinister in a 77 year old man].
We report the case of a 77 year old man, who was hospitalized in the final stadium of his congenital disease. Following a pneumonia in the right lung the respirator therapy was needed. Later the patient died by a chronic right heart failure.


Antihyperglycemic activity of L-norvaline and L-arginine in high-fat diet and streptozotocin-treated male rats.
A decrease in nitric oxide (NO) bioavailability has been shown to cause hyperglycemia, type II diabetes mellitus (DM), and chronic cardio-metabolic complications. In turn, hyperglycemia and hypercholesterolemia are associated with increased oxidative stress that leads to reduced nitric oxide bioavailability through disruption of L-arginine transport into cells, inactivation of nitric oxide synthase, and activation of arginase. Upregulation of arginase has been demonstrated in both diabetic patients and animal models of hyperglycemia and type 2 diabetes. L-norvaline is a nonselective inhibitor of arginase that increases NO production and promotes the normal functioning of the vascular endothelium. Another means of increasing NO bioavailability in the cardiovascular system is L-arginine supplementation. Whether L-norvaline and L-arginine have antihyperglycemic effects has not been studied. We hypothesized that inhibition of arginase will provide an antihyperglycemic effect and, as a result of the recovery of NO bioavailability, will protect against oxidative stress and hypercholesterolemia. Rats were fed a high-fat diet (HFD) for three weeks concomitant with the two-time injection of 30 mg/kg of streptozotocin (STZ) to induce stable hyperglycemia. We studied the antihyperglycemic properties of arginase inhibition (via L-norvaline) and its combination with NOS substrate supplementation (via L-arginine). Treatment of HFD/STZ mice with L-norvaline and L-arginine reduced fasting blood glucose levels by 27.1% vs. untreated HFD/STZ rats (p < 0.001). Blood levels of total cholesterol, low-density lipoprotein (LDL), and malondialdehyde (MDA), a marker for oxidative stress, were significantly decreased in both L-norvaline- and L-norvaline+L-arginine-treated HFD/STZ rats when compared with untreated rats. In addition, administration of L-norvaline and L-arginine reversed the progression of pancreatic and kidney pathology in HFD/STZ rats as assessed by histology (p < 0.001). Both L-norvaline and L-arginine act as potent antihyperglycemic agents and can represent alternative therapeutic tools in individuals with hyperglycemia and pre-diabetes.


Resistance to eyespot of wheat, caused by Tapesia yallundae, derived from Thinopyrum intermedium homoeologous group 4 chromosome.
Thinopyrum intermedium was identified previously as resistant to Tapesia yallundae, cause of eyespot of wheat. Using GUS-transformed isolates of T. yallundae as inoculum, we determined that wheat lines carrying Th. intermedium chromosome 4 Ai#2 or the short arm of chromosome 4 Ai#2 were as resistant to the pathogen as the eyespot-resistant wheat- Th. ponticum chromosome substitution line SS 767 (PI 611939) and winter wheat cultivar Madsen, which carries gene Pch 1 for eyespot resistance. Chromosome 4 E from Th. elongatum and chromosome 4 J from Th. bessarabicum did not confer resistance to T. yallundae. Genome-specific PCR primers confirmed the presence of Thinopyrum chromatin in these wheat- Thinopyrum lines. Genomic in situ hybridization using an St genomic probe from Pseudoroegneria strigosa demonstrated that chromosome 4 Ai#2 belongs to the J(s) genome of Thinopyrum. The eyespot resistance in the wheat- Th. intermedium lines is thus controlled by the short arm of this J(s) chromosome. This is the first report of resistance to T. yallundae controlled by a J(s) genome chromosome of Th. intermedium.


A fully protected hydrogenase/polymer-based bioanode for high-performance hydrogen/glucose biofuel cells.
Hydrogenases with Ni- and/or Fe-based active sites are highly active hydrogen oxidation catalysts with activities similar to those of noble metal catalysts. However, the activity is connected to a sensitivity towards high-potential deactivation and oxygen damage. Here we report a fully protected polymer multilayer/hydrogenase-based bioanode in which the sensitive hydrogen oxidation catalyst is protected from high-potential deactivation and from oxygen damage by using a polymer multilayer architecture. The active catalyst is embedded in a low-potential polymer (protection from high-potential deactivation) and covered with a polymer-supported bienzymatic oxygen removal system. In contrast to previously reported polymer-based protection systems, the proposed strategy fully decouples the hydrogenase reaction form the protection process. Incorporation of the bioanode into a hydrogen/glucose biofuel cell provides a benchmark open circuit voltage of 1.15 V and power densities of up to 530 µW cm-2 at 0.85 V.


Development of antibody-labelled superparamagnetic nanoparticles for the visualisation of benzo[a]pyrene in porous media with magnetic resonance imaging.
Biogeochemical interfaces in soil are dynamic in the spatial and temporal domain and require advanced visualisation and quantification tools to link in vitro experiments with natural systems. This study presents the development, characterization and application of functional nanoparticles coated with monoclonal antibodies to visualise the distribution of benzo[a]pyrene in porous media using magnetic resonance imaging. The labelled particles are 450 nm in diameter and interact with benzo[a]pyrene covalently bound to silanized silica gel. They did not bind to benzo[a]pyrene adsorbed to plain silica gel. Although unspecific filtration was low, washing steps are required for visualisation. The ability to visualise benzo[a]pyrene is inversely correlated to the heterogeneity of the soil materials. There are access restrictions to narrow pore spaces which allow the visualisation of only those pathways which are also accessible to bacteria and hydrocolloids. The production of the particles is applicable to other antibodies which extends the range of potential target contaminants.


Association of BARD1 and BRIP1 Gene Polymorphisms with the Risk of Uveal Melanoma.
Considering the limited information about the role of hereditary predisposition to the development of uveal melanoma, we have performed an analysis of the frequencies of BARD1 (rs1048108, rs2229571, rs2070094) and BRIP1 (rs4986764) gene polymorphisms in patients with uveal melanoma and benign choroidal nevus in comparison with healthy volunteers (control). It has been found that the minor alleles of BRIP1 rs4986764 and BARD1 rs2070094 polymorphisms, as well as the homozygosity of T allele at the BARD1 rs1048108 locus are common genetic markers for the predisposition to uveal melanoma and benign choroidal nevus, while the homozygous genotype GG for the BARD1 rs2229571 polymorphism is a specific marker for the predisposition to uveal melanoma and progressive choroidal nevus. We have also found that the heterozygous genotype at BARD1 rs1048108 polymorphic locus is a specific marker for protection against uveal melanoma and progressive choroidal nevus. Thus, our results indicate the advisability of studying polymorphisms of the BARD1 gene (rs1048108, rs2229571, and rs2070094) and the BRIP1 gene (rs4986764) in patients with uveal melanoma and progressive choroidal nevus. The obtained findings can be used for forming risk groups, prevention of uveal melanoma, and differential diagnosis of intraocular neoplasms.


Expression of a candidate marker for progenitor cells, Musashi-1, in the proliferative regions of human antrum and its decreased expression in intestinal metaplasia.
Reliable makers for progenitor cells in the human stomach have not been elucidated. The aim of the present study was to clarify whether Musashi-1 (Msi-1), which has recently been proposed as a stem cell marker in mouse intestine, serves as a marker for progenitor cells in human stomach. Immunohistochemistry revealed that Msi-1+ cells were detected especially in the isthmus/neck region (the putative position of stem cells) of the adult antrum, but were limited to the basal regions of fetal pyloric glands during the early stages of development. These results suggest that Msi-1 expression occurs specifically in the stem cell-containing regions. Msi-1+ cells were intermingled with proliferating cell nuclear antigen (PCNA)+ cells in the isthmus/neck region of the adult antrum, but did not coexpress PCNA or Ki 67. Msi-1 expression overlapped partly with expression of MUC 5 AC and MUC 6, indicating that Msi-1+ cells retain some features of both foveolar and pyloric gland cell differentiation phenotypes. In contrast, Msi-1 expression in gastric glands showing intestinal metaplasia (IM) became weaker than that in the glands without IM. The specific expression of Msi-1 within the proliferative regions suggests that Msi-1 is a marker of cells with progenitor characteristics before active proliferation in human antrum.


Tris(trimethylsilyl)silane as a co-initiator for dental adhesive: Photo-polymerization kinetics and dynamic mechanical property.
The purpose of this study was to evaluate the polymerization behavior of a model dentin adhesive with tris(trimethylsilyl)silane (TTMSS) as a co-initiator, and to investigate the polymerization kinetics and mechanical properties of copolymers in dry and wet conditions. A co-monomer mixture based on HEMA/BisGMA (45/55, w/w) was used as a model dentin adhesive. The photoinitiator system included camphorquinone (CQ) as the photosensitizer and the co-initiator was ethyl-4-(dimethylamino) benzoate (EDMAB) or TTMSS. Iodonium salt, diphenyliodonium hexafluorophosphate (DPIHP) serving as a catalyst, was selectively added into the adhesive formulations. The control and the experimental formulations were characterized with regard to the degree of conversion (DC) and dynamic mechanical properties under dry and wet conditions. In two-component photoinitiator system (CQ/TTMSS), with an increase of TTMSS concentration, the polymerization rate and DC of CC double bond increased, and showed a dependence on the irradiation time and curing light intensity. The copolymers that contained the three-component photoinitiator system (CQ/TTMSS/DPIHP) showed similar dynamic mechanical properties, under both dry and wet conditions, to the EDMAB-containing system. The DC of formulations using TTMSS as co-initiator showed a strong dependence on irradiation time. With the addition of TTMSS, the maximum polymerization rate can be adjusted and the network structure became more homogenous. The results indicated that the TTMSS could be used as a substitute for amine-type co-initiator in visible-light induced free radical polymerization of methacrylate-based dentin adhesives.


Acute, distribution, and subchronic toxicological studies of succinate tartrates.
The estimated single-dose oral toxicity (50% lethality) of succinate tartrates (ST) was 2-3 g/kg in rats. ST produced minimal to moderate dermal irritation but no evidence of systemic toxicity in a standard acute percutaneous toxicity test in rabbits. ST was not an eye irritant in a standard rabbit low-volume eye irritation test. ST was not genotoxic in a series of six genotoxicity tests. A 14-day oral gavage study in rats at a dose range of 0.05-1.0 g ST/kg/day produced only gastric irritation. The no-observed-effect level (NOEL) for gastric irritation was 0.1 g/kg for males and 0.05 g/kg for females. A 28-day percutaneous toxicity study in rabbits produced minimal to moderate dermal irritation and no adverse systemic effects at a high dose of 450 mg ST/kg/day. Single-dose absorption, distribution, and elimination (ADE) studies in male rats showed that 10-15% of an oral dose and 1-3% of a dermal dose were absorbed. Approximately 98% of the orally administered ST was eliminated as 14C in urine, feces, or expired CO2 after 72 hr. Approximately 80% of the dermally absorbed 14C dose was eliminated in urine, feces, or expired CO2 after 72 hr. In conclusion, no adverse effects were noted in acute toxicity, genotoxicity, or subchronic toxicity studies conducted with ST.


[Device therapy of chronic heart failure: Update 2015].
Cardiac pacemakers, implantable cardioverter defibrillators (ICD) and systems for cardiac resynchronization therapy (CRT) represent an important component of heart failure therapy. Pacemakers only play a role in bradycardia-associated heart failure and require optimal programming to prevent ventricular desynchronization. Primary prophylactic ICD implantation is indicated in patients with a left ventricular ejection fraction of ≤ 35 %, clinical stages NYHA II-III and a life expectancy > 1 year. The CRT is indicated in patients with a left bundle branch block but only in individual cases for other QRS morphologies of < 150 ms duration. The combination of CRT with a pacemaker or defibrillator must be decided on an individual basis. Device therapy in heart failure should always include remote monitoring to detect events early and to implement treatment accordingly. New developments include quadripolar left ventricular leads and pacing from multiple sites simultaneously thus enabling better resynchronization. Stimulation for modulation of cardiac contractility and the autonomous nervous system are currently being clinically tested. The optimal utilization of device therapy improves the course of heart failure and prevents cardiac decompensation and fatalities.


The genetics of a small autosomal region of Drosophila melanogaster containing the structural gene for alcohol dehydrogenase. VII. Characterization of the region around the snail and cactus loci.
The genetic interval 35C to 36A on chromosome arm 2L of Drosophila melanogaster has been saturated for mutations with visible or lethal phenotypes. 38 loci have been characterized, including several maternal-effect lethals (vasa, Bic-C, chiffon, cactus and cornichon) and several early embryonic lethals, including snail and fizzy. About 130 deletions have been used to order these loci. Complex interactions between mutant alleles have been uncovered in the immediate genetic environs of the snail gene, as has further evidence for an interaction between this region and that including the nearby genes no-ocelli and elbow.