Determination of metal-based nanoparticles in the river Dommel in the Netherlands via ultrafiltration, HR-ICP-MS and SEM.
We investigated the occurrence of metal-based nanoparticles in a natural system, the river Dommel in the Netherlands. The river itself is well-studied as far as hydrology and water quality is concerned, easily accessible and contains one major wastewater treatment plant discharging onto this river. We sampled water from various locations along the river and collected samples of influent, effluent and sewage sludge from the wastewater treatment plant. The sampling campaign was carried out in June 2015 and these samples were analysed for seven elements using high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS), ultrafiltration with a sequence of mesh sizes and scanning electron microscopy (SEM). From the results we conclude that there are indeed nanoparticles present in the treatment plant we studied, as we found titanium and gold particles in the influent and effluent. In the river water only 10 to 20% of the mass concentration of titanium, cerium and other elements we examined is made up of free, i.e. unattached, particles with a size smaller than 20 nm or of dissolved material. The rest is attached to natural colloids or is present as individual particles or clusters of smaller particles, as it could be filtered out with 450 nm ultrafilters. We found evidence that there is no appreciable anthropogenic emission of cerium into the river, based on the geochemical relationship between cerium and lanthanum. Besides, the effluent of the treatment plant has lower concentrations of some examined elements than the surface water upstream. The treatment plant discharges much less of these elements than estimated using previous publications. However, a potential diffuse source of titanium dioxide in the form of nanoparticles or of larger particles is their use in paints and coatings, as the concentration of titanium increased considerably in the urbanised area of the river Dommel.


Mushroom biomass and diversity are driven by different spatio-temporal scales along Mediterranean elevation gradients.
Mushrooms are important non-wood-forest-products in many Mediterranean ecosystems, being highly vulnerable to climate change. However, the ecological scales of variation of mushroom productivity and diversity, and climate dependence has been usually overlooked due to a lack of available data. We determined the spatio-temporal variability of epigeous sporocarps and the climatic factors driving their fruiting to plan future sustainable management of wild mushrooms production. We collected fruiting bodies in Pinus sylvestris stands along an elevation gradient for 8 consecutive years. Overall, sporocarp biomass was mainly dependent on inter-annual variations, whereas richness was more spatial-scale dependent. Elevation was not significant, but there were clear elevational differences in biomass and richness patterns between ectomycorrhizal and saprotrophic guilds. The main driver of variation was late-summer-early-autumn precipitation. Thus, different scale processes (inter-annual vs. spatial-scale) drive sporocarp biomass and diversity patterns; temporal effects for biomass and ectomycorrhizal fungi vs. spatial scale for diversity and saprotrophic fungi. The significant role of precipitation across fungal guilds and spatio-temporal scales indicates that it is a limiting resource controlling sporocarp production and diversity in Mediterranean regions. The high spatial and temporal variability of mushrooms emphasize the need for long-term datasets of multiple spatial points to effectively characterize fungal fruiting patterns.


Deletion of Bmal1 Prevents Diet-Induced Ectopic Fat Accumulation by Controlling Oxidative Capacity in the Skeletal Muscle.
Brain and muscle arnt-like protein 1 (BMAL1), is a transcription factor known to regulate circadian rhythm. BMAL1 was originally characterized by its high expression in the skeletal muscle. Since the skeletal muscle is the dominant organ system in energy metabolism, the possible functions of BMAL1 in the skeletal muscle include the control of metabolism. Here, we established that its involvement in the regulation of oxidative capacity in the skeletal muscle. Muscle-specific Bmal1 KO mice (MKO mice) displayed several physiological hallmarks for the increase of oxidative capacity. This included increased energy expenditure and oxygen consumption, high running endurance and resistance to obesity with improved metabolic profiles. Also, the phosphorylation status of AMP-activated protein kinase and its downstream signaling substrate acetyl-CoA carboxylase in the MKO mice were substantially higher than those in the Bmal1flox/flox mice. In addition, biochemical and histological studies confirmed the substantial activation of oxidative fibers in the skeletal muscle of the MKO mice. The mechanism includes the regulation of Cacna1s expression, followed by the activation of calcium-nuclear factor of activated T cells (NFAT) axis. We thus conclude that BMAL1 is a critical regulator of the muscular fatty acid level under nutrition overloading and that the mechanism involves the control of oxidative capacity.


Dietary exposure to metals and other elements in the 2006 UK Total Diet Study and some trends over the last 30 years.
Concentrations of 24 elements including metals in the 2006 UK Total Diet Study (TDS) were measured and dietary exposures estimated. Composite samples for the 20 TDS food groups (bread, fish, fruit, etc.) were collected from 24 UK towns and analysed for their levels of aluminium, antimony, arsenic, barium, bismuth, cadmium, chromium, copper, germanium, indium, lead, manganese, mercury, molybdenum, nickel, palladium, platinum, rhodium, ruthenium, selenium, strontium, thallium, tin, and zinc. Concentrations of each of the elements in the food groups were lower than or similar to those reported in the previous TDS survey, conducted in 2000, with the exception of aluminium, barium, and manganese. Dietary exposures to the 24 elements were estimated for UK consumers and compared with previous estimates made over the last 30 years in order to examine any trends in exposure to these elements in the typical UK diet. Population exposures to the elements have generally declined over time, and exposures to most of these elements remain at low levels. The independent UK Government scientific Committee on Toxicity of Chemicals in Food, Consumer Products and the Environment (COT) commented on the estimated dietary exposures, taking into account their previous evaluations (in 2003 and 2008), and identified no major concerns for the health of consumers, but did advise that there was a need for more information on aluminium and barium, and also commented that dietary exposure to inorganic arsenic and to lead should continue to be reduced.


Piriformis syndrome: a case series of 31 Bangladeshi people with literature review.
To describe a series of piriformis syndrome patient among Bangladesh people with literature review. Consecutive 31 piriformis syndrome patients were enrolled. Besides history and clinical examination, piriformis muscle thickness was also measured with diagnostic ultrasound (3.5 MHZ). MRI of lumbar spine, X-rays of lumbo-sacral spine, and pelvis were performed in all patients. Statistical Package for the Social Sciences (SPSS), Windows 8.0, was used for statistical calculation, and univariate analysis of primary data was done. Data present with frequency table. For literature review concerning piriformis syndrome we used Embase, Pubmed, Medline, and Cochrane database. A total of 31 patients (21 female) with PS were enrolled, 21 housewives. Mean age 42.2 ± 14.5 years. All presented with buttock pain, aggravating with long sitting (31), lying on the affected side (31), during rising from a chair(24), and forward bending (28). Six reported pain improvement while walking. Gluteal tenderness, positive FAIR test, and Pace sign were elicited in all patients. A palpable gluteal mass was found in 8 cases, gluteal atrophy in 5 other patients. The mean piriformis muscle thickness on the diseased side was more than on the healthy side (13.6 ± 3.7 vs 10.9 ± 1.9, (p > 0.05). Common conditions associated with PS were: preceding fall (9, 29 %), overuse of piriformis muscle, lumbar spinal stenosis, fibromyalgia, intra-muscular gluteal injection, blunt trauma over the buttock, leg length discrepancy and use of rear pocket's wallet. In Bangladesh piriformis syndrome is more common in female, especially among housewives. A fall often precedes the condition. Piriformis syndrome should be considered as possible diagnosis when sciatica occurs without a clear spine pathology.


Low Fe(II) Concentrations Catalyze the Dissolution of Various Fe(III) (hydr)oxide Minerals in the Presence of Diverse Ligands and over a Broad pH Range.
Dissolution of Fe(III) (hydr)oxide minerals by siderophores (i.e., Fe-specific, biogenic ligands) is an important step in Fe acquisition in environments where Fe availability is low. The observed coexudation of reductants and ligands has raised the question of how redox reactions might affect ligand-controlled (hydr)oxide dissolution and Fe acquisition. We examined this effect in batch dissolution experiments using two structurally distinct ligands (desferrioxamine B (DFOB) and N, N'-di(2-hydroxybenzyl)ethylene-diamine- N, N'-diacetic acid (HBED)) and four Fe(III) (hydr)oxide minerals (lepidocrocite, 2-line ferrihydrite, goethite and hematite) over an environmentally relevant pH range (4-8.5). The experiments were conducted under anaerobic conditions with varying concentrations of (adsorbed) Fe(II) as the reductant. We observed a catalytic effect of Fe(II) on ligand-controlled dissolution even at submicromolar Fe(II) concentrations with up to a 13-fold increase in dissolution rate. The effect was larger for HBED than for DFOB. It was observed for all four Fe(III) (hydr)oxide minerals, but it was most pronounced for goethite in the presence of HBED. It was observed over the entire pH range with the largest effect at pH 7 and 8.5, where Fe deficiency typically occurs. The occurrence of this catalytic effect over a range of environmentally relevant conditions and at very low Fe(II) concentrations suggests that redox-catalyzed, ligand-controlled dissolution may be significant in biological Fe acquisition and in redox transition zones.


Temporal Tracking of Microglia Activation in Neurodegeneration at Single-Cell Resolution.
Microglia, the tissue-resident macrophages in the brain, are damage sensors that react to nearly any perturbation, including neurodegenerative diseases such as Alzheimer's disease (AD). Here, using single-cell RNA sequencing, we determined the transcriptome of more than 1,600 individual microglia cells isolated from the hippocampus of a mouse model of severe neurodegeneration with AD-like phenotypes and of control mice at multiple time points during progression of neurodegeneration. In this neurodegeneration model, we discovered two molecularly distinct reactive microglia phenotypes that are typified by modules of co-regulated type I and type II interferon response genes, respectively. Furthermore, our work identified previously unobserved heterogeneity in the response of microglia to neurodegeneration, discovered disease stage-specific microglia cell states, revealed the trajectory of cellular reprogramming of microglia in response to neurodegeneration, and uncovered the underlying transcriptional programs.


Midazolam versus diazepam for the treatment of status epilepticus in children and young adults: a meta-analysis.
Rapid treatment of status epilepticus (SE) is associated with better outcomes. Diazepam and midazolam are commonly used, but the optimal agent and administration route is unclear. The objective was to determine by systematic review if nonintravenous (non-IV) midazolam is as effective as diazepam, by any route, in terminating SE seizures in children and adults. Time to seizure cessation and respiratory complications was examined. We performed a search of PubMed, Web of Knowledge, Embase, Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects, American College of Physicians Journal Club, Cochrane Central Register of Controlled Trials, the Cumulative Index to Nursing and Allied Health Literature, and International Pharmaceutical Abstracts for studies published January 1, 1950, through July 4, 2009. English language quasi-experimental or randomized controlled trials comparing midazolam and diazepam as first-line treatment for SE, and meeting the Consolidated Standards of Reporting Trials (CONSORT)-based quality measures, were eligible. Two reviewers independently screened studies for inclusion and extracted outcomes data. Administration routes were stratified as non-IV (buccal, intranasal, intramuscular, rectal) or IV. Fixed-effects models generated pooled statistics. Six studies with 774 subjects were included. For seizure cessation, midazolam, by any route, was superior to diazepam, by any route (relative risk [RR] = 1.52; 95% confidence interval [CI] = 1.27 to 1.82). Non-IV midazolam is as effective as IV diazepam (RR = 0.79; 95% CI = 0.19 to 3.36), and buccal midazolam is superior to rectal diazepam in achieving seizure control (RR = 1.54; 95% CI = 1.29 to 1.85). Midazolam was administered faster than diazepam (mean difference = 2.46 minutes; 95% CI = 1.52 to 3.39 minutes) and had similar times between drug administration and seizure cessation. Respiratory complications requiring intervention were similar, regardless of administration route (RR = 1.49; 95% CI = 0.25 to 8.72). Non-IV midazolam, compared to non-IV or IV diazepam, is safe and effective in treating SE. Comparison to lorazepam, evaluation in adults, and prospective confirmation of safety and efficacy is needed.


Acute serious rebleeding after angiographically successful coil embolization of ruptured cerebral aneurysms.
The present study investigated the incidence of acute rebleeding after successful coil embolization of a ruptured cerebral aneurysm, including clinical outcomes, and possible mechanisms of the events other than coil compaction and/or incomplete embolization. This study included 591 consecutive patients who presented with aneurysmal subarachnoid hemorrhage, were treated with coil embolization, and whose post-procedural angiography revealed successful embolization. Data were collected retrospectively from six patients who showed acute rebleeding despite that angiographically successful coil embolization was achieved. All clinical, radiological data and intraoperative videos were reviewed to identify causative factors which could have contributed to the occurrence of rebleeding. Incidence of acute rebleeding after successful coil embolization of ruptured cerebral aneurysm was 1.0% (6/591). In all of these six patients, complete angiographic occlusion was achieved except in one case where a small residual neck was intentionally left to avoid compromise of the parent artery. Four of the six patients showed poor clinical courses, either died or recovered with severe disability. Whenever possible, we performed an immediate craniotomy for exploration and additional clipping. Based on intraoperative findings, we hypothesized that uneven distribution of the coil masses and spontaneous resolution of thrombus among the strands of coil (inter-coil-loop thrombolysis) could be possible mechanisms of rebleeding. Acute rebleeding is extremely rare, but is possible as a complication of coil embolization of a ruptured cerebral aneurysm even when a case is angiographically successful. The higher degree of morbidity and mortality is a major concern. Therefore, further investigation to discover risk factors and causative mechanisms for such a complication is sorely needed.


Prescribers' perceptions of benefits and limitations of direct acting oral anticoagulants in non-valvular atrial fibrillation.
There is an acknowledged lack of robust and rigorous research focusing on the perspectives of those prescribing direct acting oral anticoagulants (DOACs) for non-valvular atrial fibrillation (AF). The objective was to describe prescribers' experiences of using DOACs in the management of non-valvular AF, including perceptions of benefits and limitations. A cross-sectional survey of prescribers in a remote and rural area of Scotland. Among other items, the questionnaire invited free-text description of positive and negative experiences of DOACs, and benefits and limitations. Responses were independently analysed by two researchers using a summative content analysis approach. This involved counting and comparison, via keywords and content, followed by interpretation and coding of the underlying context into themes. One hundred and fifty-four responses were received, 120 (77.9%) from physicians, 18 (11.7%) from nurse prescribers and 10 (6.4%) from pharmacist prescribers (6 unidentified professions). Not having to monitor INR was the most cited benefit, particularly for prescribers and patients in remote and rural settings, followed by potentially improved patient adherence. These benefits were reflected in respondents' descriptions of positive experiences and patient feedback. The main limitations were the lack of reversal agents, cost and inability to monitor anticoagulation status. Many described their experiences of adverse effects of DOACs including fatal and non-fatal bleeding, and upper gastrointestinal disturbances. While prescribers have positive experiences and perceive benefits of DOACs, issues such as adverse effects and inability to monitor anticoagulation status merit further monitoring and investigation. These issues are particularly relevant given the trajectory of increased prescribing of DOACs.


Targeting the PELP1-KDM1 axis as a potential therapeutic strategy for breast cancer.
The estrogen receptor (ER) co-regulator proline glutamic acid and leucine-rich protein 1 (PELP1) is a proto-oncogene that modulates epigenetic changes on ER target gene promoters via interactions with lysine-specific histone demethylase 1 (KDM1). In this study, we assessed the therapeutic potential of targeting the PELP1-KDM1 axis in vivo using liposomal (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine; DOPC) siRNA to downregulate PELP1 expression and KDM1 inhibitors, pargyline and N-((1S)-3-(3-(trans-2-aminocyclopropyl)phenoxy)-1-(benzylcarbamoyl)propyl)benzamide using preclinical models. Preclinical xenograft models were used to test the efficacy of drugs in vivo. Ki-67 and terminal deoxynucleotidyl transferase dUTP nick end-labeling immunohistochemical analysis of epigenetic markers was performed on tumor tissues. The in vitro effect of PELP1-KDM axis blockers was tested using proliferation, reporter gene, chromatin immunoprecipitation and real-time RT-PCR assays. The efficacy of the KDM1 targeting drugs alone or in combination with letrozole and tamoxifen was tested using therapy-resistant model cells. Treatment of ER-positive xenograft-based breast tumors with PELP1-siRNA-DOPC or pargyline reduced tumor volume by 58.6% and 62%, respectively. In a postmenopausal model, in which tumor growth is stimulated solely by local estrogen synthesis, daily pargyline treatment reduced tumor volume by 78%. Immunohistochemical analysis of excised tumors revealed a combined decrease in cellular proliferation, induction of apoptosis and upregulation of inhibitory epigenetic modifications. Pharmacological inhibition of KDM1 in vitro increased inhibitory histone mark dimethylation of histone H3 at lysine 9 (H3K9me2) and decreased histone activation mark acetylation of H3K9 (H3K9Ac) on ER target gene promoters. Combining KDM1 targeting drugs with current endocrine therapies substantially impeded growth and restored sensitivity of therapy-resistant breast cancer cells to treatment. Our results suggest inhibition of PELP1-KDM1-mediated histone modifications as a potential therapeutic strategy for blocking breast cancer progression and therapy resistance.


Computer-assisted analysis and epidemiological value of genotyping methods for Campylobacter jejuni and Campylobacter coli.
For epidemiological tracing of the thermotolerant Campylobacter species C. jejuni and C. coli, reliable and highly discriminatory typing techniques are necessary. In this study the genotyping techniques of flagellin typing (flaA typing), pulsed-field gel electrophoresis (PFGE), automated ribotyping, and amplified fragment length polymorphism (AFLP) fingerprinting were compared. The following aspects were compared: computer-assisted analysis, discriminatory power, and use for epidemiological typing of campylobacters. A set of 50 campylobacter poultry isolates from The Netherlands and neighboring countries was analyzed. Computer-assisted analysis made cluster analysis possible and eased the designation of different genotypes. AFLP fingerprinting was the most discriminatory technique, identifying 41 distinct genotypes, while PFGE identified 38 different types, flaA typing discriminated 31 different types, and ribotyping discriminated 26 different types. Furthermore, AFLP analysis was the most suitable method for computer-assisted data analysis. In some cases combining the results of AFLP fingerprinting, PFGE, and flaA typing increased our ability to differentiate strains that appeared genetically related. We conclude that AFLP is a highly discriminatory typing method and well suited for computer-assisted data analysis; however, for optimal typing of campylobacters, a combination of multiple typing methods is needed.


[Individual resistance to factors of monotony reflected in the EEG].
Three groups of operators substantially different in their performance quality were examined. Under conditions of monotonous activity, subjects with the highest initial level of activation, minimum EEG total spectral power, and minimum level of EEG coherence in the frontal cortical areas worked most steadily. Under the same conditions, subjects with a rather high spectral power of the theta and beta2 EEG frequency components, highest coherence in the frontal areas, and low coherence in the caudal areas of the cortex worked least steadily. EEG phenomena testify to a rather low level of activation of the frontal cortical areas associated with a facilitation of cortico-subcortical neuronal interactions and an attenuation of the operating neural streams. This results in a decrease in the level of any attention, its involuntary switching, and short-term loss of the control over the current performance.


Communication gaps and readmissions to hospital for patients aged 75 years and older: observational study.
(1) To identify communication gaps at hospital discharge for older people who are readmitted within 28 days; (2) to assess the contribution of incomplete discharge information to readmission; (3) to identify measures that might improve communication at hospital discharge. Retrospective case-note review. A teaching hospital in Nottingham, UK. 108 consecutive patients aged 75 and over who were readmitted as an emergency within 28 days of previous discharge (1) Proportion of patients discharged with insufficient arrangements and/or information for immediate safe continuity of care; (2) proportion of patients with medication management information missing; (3) proportion of readmissions related to incomplete medication management information and proportion of patients for whom this was preventable. Thirty patients (28%) returned within 3 days of discharge, 48 (44%) within 7 days and the remainder within 28 days. Sixty-seven (62%) patients either had no discharge letter or returned before the letter was typed. Documentation of changes in medication was incomplete on two-thirds of all discharge documents. Readmission was considered to be related to medication for 41 (38%) patients and to be preventable for 25 (61%) of these. There were preventable discharge communication gaps, including monitoring information, for 22 (54%) of these patients. Incomplete documentation at discharge was common, particularly for medication management. It is likely that communication gaps contributed to many of the preventable adverse events and readmissions.


Potent induction of total cellular and mitochondrial antioxidants and phase 2 enzymes by cruciferous sulforaphane in rat aortic smooth muscle cells: cytoprotection against oxidative and electrophilic stress.
Sulforaphane, a cruciferous isothiocyanate compound, upregulates cytoprotective genes in liver, but its effects on antioxidants and phase 2 defenses in vascular cells are unknown. Here we report that incubation of rat aortic smooth muscle A10 cells with sulforaphane (0.25-5 microM) resulted in concentration-dependent induction of a spectrum of important cellular antioxidants and phase 2 enzymes, including superoxide dismutase (SOD), catalase, the reduced form of glutathione (GSH), glutathione peroxidase, glutathione reductase (GR), glutathione S-transferase (GST), and NAD(P)H:quinone oxidoreductase 1 (NQO1). Sulforaphane also increased levels/activities of SOD, catalase, GSH and GST in isolated mitochondria of aortic smooth muscle cells. Time-dependent sulforaphane-induced increases in the mRNA levels for MnSOD, catalase, the catalytic subunit of gamma-glutamylcysteine ligase, GR, GST-A1, GST-P1, and NQO1 were observed. Pretreatment with sulforaphane (0.5, 1, and 5 microM) protected aortic smooth muscle cells from oxidative and electrophilic cytotoxicity induced by xanthine oxidase (XO)/xanthine, H2O2, SIN-1-derived peroxynitrite, 4-hydroxy-2-nonenal, and acrolein. Furthermore, sulforaphane pretreatment prevented intracellular accumulation of reactive oxygen species (ROS) after exposure of the cells to XO/xanthine, H2O2, or SIN-1. Taken together, this study demonstrates that in the aortic smooth muscle cells sulforaphane at physiologically relevant concentrations potently induces a series of total cellular as well as mitochondrial antioxidants and phase 2 enzymes, which is accompanied by dramatically increased resistance of these vascular cells to oxidative and electrophilic stress.


A study of the influence of sex on genome wide methylation.
Sex differences in methylation status have been observed in specific gene-disease studies and healthy methylation variation studies, but little work has been done to study the impact of sex on methylation at the genome wide locus-to-locus level or to determine methods for accounting for sex in genomic association studies. In this study we investigate the genomic sex effect on saliva DNA methylation of 197 subjects (54 females) using 20,493 CpG sites. Three methods, two-sample T-test, principle component analysis and independent component analysis, all successfully identify sex influences. The results show that sex not only influences the methylation of genes in the X chromosome but also in autosomes. 580 autosomal sites show strong differences between males and females. They are found to be highly involved in eight functional groups, including DNA transcription, RNA splicing, membrane, etc. Equally important is that we identify some methylation sites associated with not only sex, but also other phenotypes (age, smoking and drinking level, and cancer). Verification was done through an independent blood cell DNA methylation data (1298 CpG sites from a cancer panel array). The same genomic site-specific influence pattern and potential confounding effects with cancer were observed. The overlapping rate of identified sex affected genes between saliva and blood cell is 81% for X chromosome, and 8% for autosomes. Therefore, correction for sex is necessary. We propose a simple correction method based on independent component analysis, which is a data driven method and accommodates sample differences. Comparison before and after the correction suggests that the method is able to effectively remove the potentially confounding effects of sex, and leave other phenotypes untouched. As such, our method is able to disentangle the sex influence on a genome wide level, and paves the way to achieve more accurate association analyses in genome wide methylation studies.


Radiographic method for anatomic study of the teat canal: changes between milking periods.
Radiographs of the teat canal (papillary duct) of 14 Holstein-Friesian cows were made immediately after removal of the milking machine and at 2, 4, 6, and 8 hours. The distal, middle, and proximal diameters of the canal were dilated when the machine was removed. The distal and the middle parts of the teat canal were more dilated than the proximal part of the canal. Two hours later, the canal was constricted in the 3 areas. During the next 6 hours, the canal dilated-the greatest kilation being at the proximal paet of the teat canal. The average lenght of the teat canal remained unchanged. Apparently, microorganisms can easily pass through the teat canal when it is dilated at the end of machine milking and for up to 2 hours after milking.


Translation Inhibition by Rocaglates Activates a Species-Specific Cell Death Program in the Emerging Fungal Pathogen Candida auris.
Fungal infections are a major contributor to infectious disease-related deaths worldwide. Recently, global emergence of the fungal pathogen Candida auris has caused considerable concern because most C. auris isolates are resistant to fluconazole, the most commonly administered antifungal, and some isolates are resistant to drugs from all three major antifungal classes. To identify novel agents with bioactivity against C. auris, we screened 2,454 compounds from a diversity-oriented synthesis collection. Of the five hits identified, most shared a common rocaglate core structure and displayed fungicidal activity against C. auris These rocaglate hits inhibited translation in C. auris but not in its pathogenic relative Candida albicans Species specificity was contingent on variation at a single amino acid residue in Tif1, a fungal member of the eukaryotic initiation factor 4A (eIF4A) family of translation initiation factors known to be targeted by rocaglates. Rocaglate-mediated inhibition of translation in C. auris activated a cell death program characterized by loss of mitochondrial membrane potential, increased caspase-like activity, and disrupted vacuolar homeostasis. In a rocaglate-sensitized C. albicans mutant engineered to express translation initiation factor 1 (Tif1) with the variant amino acid that we had identified in C. auris, translation was inhibited but no programmed cell death phenotypes were observed. This surprising finding suggests divergence between these related fungal pathogens in their pathways of cellular responses to translation inhibition. From a therapeutic perspective, the chemical biology that we have uncovered reveals species-specific vulnerability in C. auris and identifies a promising target for development of new, mechanistically distinct antifungals in the battle against this emerging pathogen.IMPORTANCE Emergence of the fungal pathogen Candida auris has ignited intrigue and alarm within the medical community and the public at large. This pathogen is unusually resistant to antifungals, threatening to overwhelm current management options. By screening a library of structurally diverse molecules, we found that C. auris is surprisingly sensitive to translation inhibition by a class of compounds known as rocaglates (also known as flavaglines). Despite the high level of conservation across fungi in their protein synthesis machinery, these compounds inhibited translation initiation and activated a cell death program in C. auris but not in its relative Candida albicans Our findings highlight a surprising divergence across the cell death programs operating in Candida species and underscore the need to understand the specific biology of a pathogen in attempting to develop more-effective treatments against it.


Changes in preschool children's oral health-related quality of life following restorative dental general anaesthesia.
Background Early childhood caries is the most common childhood disease in preschool children which often requires dental treatment under restorative dental general anaesthesia (RDGA).Aim To assess the effect of RDGA on preschool children and their families' oral health-related quality of life (OHRQoL).Methods A cross-sectional study using a RDGA pre- and post-operative survey to evaluate changes in OHRQoL. Parents completed the Early Childhood Oral Health Impact Scale (ECOHIS) survey before and after their children underwent specialist paediatric RDGA in the United Arab Emirates from 1 March 2017 to 28 February 2018. The ECOHIS and its effect size (ES) served to evaluate children's OHRQoL status and changes, and the Wilcoxon signed-rank and the Kruskal-Wallis tests were used for statistical analyses.Results The mean number of decayed, missing and filled teeth (DMFT) of the children (N = 173, mean age 4.6 years) before the treatment was very high (13.8 ± 3.07). Children's pain and eating problems, and parents feeling upset and guilty, were the most frequently reported impacts at baseline. The ECOHIS scores decreased significantly (p <0.001) after RDGA, revealing a large ES for the child (2.19) and family (2.79) sections of the ECOHIS.Conclusions Preschool children's RDGA resulted in significant improvement in all child and family physical, psychological and social aspects of OHRQoL.


Anti-diabetic effect of dietary mango (Mangifera indica L.) peel in streptozotocin-induced diabetic rats.
In the present study, the composition of mango peel powder (MPP) collected from the mango pulp industry was determined and the effect of MPP on ameliorating diabetes and its associated complications was studied. Mango peel was rich in polyphenols, carotenoids and dietary fibre. Peel extract contained various bioactive compounds and was found to be rich in soluble dietary fibre. Peel extract exhibited antioxidant properties and protected against DNA damage. Therefore, the effect of peel on ameliorating diabetes was investigated in a rat model of diabetes. A significant increase in urine sugar, urine volume, fasting blood glucose, total cholesterol, triglycerides and low density lipoprotein, and decrease in high density lipoprotein were observed in the rats; however, these parameters were ameliorated in diabetic rats fed with diet supplemented with mango peel at 5% and 10% levels in basal diet. Treatment of diabetic rats with MPP increased antioxidant enzyme activities and decreased lipid peroxidation in plasma, kidney and liver compared to untreated diabetic rats. Glomerular filtration rate and microalbuminuria levels were ameliorated in MPP treated diabetic group. Mango peel, a by-product, can be used as an ingredient in functional and therapeutic foods.


Localized scleroderma in childhood is not just a skin disease.
Juvenile localized scleroderma is usually considered a disease that is confined to the skin and subcutaneous tissue. We studied the prevalence and clinical features of extracutaneous manifestations in a large cohort of children with juvenile localized scleroderma. Data from a multinational study on juvenile scleroderma was used for this in-depth study. Clinical features of patients with extracutaneous manifestations were compared with those of patients who had exclusively skin involvement. Seven hundred fifty patients entered the study. One hundred sixty-eight patients (22.4%) presented with a total of 193 extracutaneous manifestations, as follows: articular (47.2%), neurologic (17.1%), vascular (9.3%), ocular (8.3%), gastrointestinal (6.2%), respiratory (2.6%), cardiac (1%), and renal (1%). Other autoimmune conditions were present in 7.3% of patients. Neurologic involvement consisted of epilepsy, central nervous system vasculitis, peripheral neuropathy, vascular malformations, headache, and neuroimaging abnormalities. Ocular manifestations were episcleritis, uveitis, xerophthalmia, glaucoma, and papilledema. In more than one-fourth of these children, articular, neurologic, and ocular involvements were unrelated to the site of skin lesions. Raynaud's phenomenon was reported in 16 patients. Respiratory involvement consisted essentially of restrictive lung disease. Gastrointestinal involvement was reported in 12 patients and consisted exclusively of gastroesophageal reflux. Thirty patients (4%) had multiple extracutaneous features, but systemic sclerosis (SSc) developed in only 1 patient. In patients with extracutaneous involvement, the prevalence of antinuclear antibodies and rheumatoid factor was significantly higher than that among patients with only skin involvement. However, Scl-70 and anticentromere, markers of SSc, were not significantly increased. Extracutaneous manifestations of juvenile localized scleroderma developed in almost one-fourth of the children in this study. These extracutaneous manifestations often were unrelated to the site of the skin lesions and sometimes were associated with multiple organ involvement. The risk of developing SSc was very low. This subgroup of patients with juvenile localized scleroderma should be evaluated extensively, treated more aggressively, and monitored carefully.


LFA-1 expression by blood eosinophils is increased in atopic asthmatic children and is involved in eosinophil locomotion.
Allergic asthma is characterized by eosinophil migration in the airways, which is strictly dependent on the expression of adhesion molecules. This study investigated whether the expression of adhesion molecules on eosinophils is increased and associated with disease activity in allergic asthma. Twenty atopic asthmatic (AA) subjects and nine controls were studied and the expression of lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18), Mac-1 (CD11b/CD18) and very late antigen-4 (VLA-4; CD49d/CD29) on blood eosinophils was evaluated by specific monoclonal antibody (Mab) staining and flow-cytometric analysis. Compared with controls, eosinophils from AA showed increased expression of LFA-1 (p<0.005), but not of Mac-1 or VLA-4 (p>0.1). In addition, LFA-1 expression correlated positively with blood eosinophil number (r=0.792, p<0.05), while no correlations were observed between Mac-1 or VLA-4 expression and blood eosinophil number. The migration of eosinophils through human umbilical vein endothelial cells with or without anti-LFA-1, Mac-1 and VLA-4-blocking Mab was studied. Compared with controls, eosinophils from AA showed increased migration toward C5a (p<0.01). Cell migration was totally inhibited by preincubating eosinophils with anti-LFA-1 (p<0.05), while anti-Mac-1 had no effect (p>0.1). Thus, the expression of lymphocyte function-associated antigen-1 by blood eosinophils is increased in atopic asthmatics and seems to modulate the enhanced eosinophil migration observed in allergic asthma.


[Biogenic amines in rat brain structures in an adjuvant arthritis model].
The role of changes in biogenic amines neurochemistry in cerebral structures in abnormal immunoreactivity accompanying development of autoimmune disease was investigated in experiments on 24 Wistar male rats. Biogenic amines and their metabolites (dopamine, dioxyphenylacetic acid, serotonin, oxyindolacetic acid) were assays in the sensomotor cortex, hypothalamus and hippocamp by means of high-performance liquid chromatography. Development of adjuvant arthritis was associated with definite neurochemical changes in the brain: inhibition of serotonin metabolism and enhancement of dopaminergic systems in the hypothalamus and sensomotor cortex. Prophylactic administration of oil solution D,L-alpha-tocopherol acetate promoted enhancement of serotonin metabolism in hypothalamic and hippocampal neurons. Bitemporal exposure to UHF electric field aroused stronger effects on the neurochemical metabolism in hypothalamic, hippocampal and sensomotor cortex neurons than TPA. It is concluded that bitemporal exposure to UHF electric field and antioxidant activate central immunoreactivity mechanisms in animals with adjuvant arthritis. Serotonin and dopamine systems in this process are leading.


Exploration and exploitation during sequential search.
When we learn how to throw darts we adjust how we throw based on where the darts stick. Much of skill learning is computationally similar in that we learn using feedback obtained after the completion of individual actions. We can formalize such tasks as a search problem; among the set of all possible actions, find the action that leads to the highest reward. In such cases our actions have two objectives: we want to best utilize what we already know (exploitation), but we also want to learn to be more successful in the future (exploration). Here we tested how participants learn movement trajectories where feedback is provided as a monetary reward that depends on the chosen trajectory. We mathematically derived the optimal search policy for our experiment using decision theory. The search behavior of participants is well predicted by an ideal searcher model that optimally combines exploration and exploitation.


Slope efficiency over 30% single-frequency ytterbium-doped fiber laser based on Sagnac loop mirror filter.
A high-slope-efficiency single-frequency (SF) ytterbium-doped fiber laser, based on a Sagnac loop mirror filter (LMF), was demonstrated. It combined a simple linear cavity with a Sagnac LMF that acted as a narrow-bandwidth filter to select the longitudinal modes. And we introduced a polarization controller to restrain the spatial hole burning effect in the linear cavity. The system could operate at a stable SF oscillating at 1064 nm with the obtained maximum output power of 32 mW. The slope efficiency was found to be primarily dependent on the reflectivity of the fiber Bragg grating. The slope efficiency of multi-longitudinal modes was higher than 45%, and the highest slope efficiency of the single longitudinal mode we achieved was 33.8%. The power stability and spectrum stability were <2% and <0.1%, respectively, and the signal-to-noise ratio measured was around 60 dB.


Quality of life and social integration of severely mentally ill patients: a longitudinal study.
The quality of life concept serves to measure functional changes and program outcome. Patients with schizophrenia have an improved prognosis. Is quality of life improving over time, and if so, over what period? These questions were addressed in a longitudinal study where subjective quality of life (SQOL) was rated by severely mentally ill patients living in the community and using support services located in an outlying area of Quebec. The Satisfaction for Life Domains Scale (SLDS) (Baker and Intagliata 1982) measuring SQOL as a whole and in specific domains (for example, housing, finances, social relationships) was repeated over a period of 7 y. Results show that SQOL ratings received the same scores after 7 y. Functional status was decreased, while social integration improved and more services were used. The results could be due to sample characteristics or to the ambiguity of the SQOL construct. In our opinion, extensive community-based support services may have played a key role in the maintenance of patient's quality of life.


Auditing stillbirths at Lower Umfolozi War Memorial Regional Hospital: A 12-month review.
Although the total number of stillbirths worldwide was estimated at 2.6 million in 2009, there is currently a dearth of literature on stillbirths in developing countries and rural settings, where the majority of such births occur. The 'Hands Up' Mortality and Morbidity Extraction Tool (HUMMET), developed at Lower Umfolozi War Memorial Regional Hospital (LUWMRH) in 2010, outlines a systematic approach to summarising individual cases of adverse perinatal outcomes. To depict the HUMMET form by describing the detailed demographic and obstetric profile of patients who delivered a stillborn infant at LUWMRH, as well as risk factors associated with these stillbirths between 1 April 2014 and 31 March 2015. The findings add to a global initiative advanced by the Lancet series on stillbirths, aimed at raising awareness of stillbirth statistics in low- and middle-income countries. A total of 310 detailed stillbirth case summaries of 305 patients were collected during the study period, representing 90% of the total number of stillborn infants delivered at LUWMRH. A retrospective audit of the HUMMET forms was conducted and the cases were further summarised in a Microsoft Excel spreadsheet that allowed for a univariate analysis of the variables. The stillbirth rate at LUWMRH is much higher than that at other regional hospitals owing to the number of at-risk referrals and emergency cases from surrounding clinics and district hospitals. Referrals were from local clinics (49%) and district hospitals (45%), 35% of stillbirths were due to abruptio placentae and a large proportion were associated with gestational hypertension, pre-eclampsia and/or eclampsia. Avoidable factors were predominantly a late patient response to reduced fetal movements and delays in transfer to hospital. Twenty percent of stillbirths were associated with inappropriate monitoring or management of the obstetric condition at the district hospital. The HUMMET form provides a systematic approach to analysing cases of perinatal morbidity and mortality in line with the requirements of the Perinatal Problem Identification Programme database, but provides more details on the circumstances and contributing factors. A repeat audit is recommended to determine whether interventions have been effective.


Granular cells are required for encapsulation of foreign targets by insect haemocytes.
Haemocytes play an essential role in defending invertebrates against pathogens and parasites that enter their haemocoel. A primary defense response is encapsulation; a process in which haemocytes attach to the foreign organism and kill it. Whether encapsulation requires cooperation between specific subpopulations of haemocytes is unknown. Using purified subpopulations of haemocytes and an in vitro encapsulation assay, we investigated the process of capsule formation in the insect Pseudoplusia includens. Immunocytochemical staining revealed that capsule formation involves a three step process. Encapsulation began when granular cells attached to the foreign target. This was followed by attachment of multiple layers of plasmatocytes. Termination of capsule formation occurred when a subpopulation of granular cells formed a monolayer around the periphery of the capsule. Neither granular cells nor plasmatocytes were capable of forming a capsule independently. However, plasmatocytes encapsulated targets if granular cells were present or if targets were preincubated in medium conditioned by granular cells. The effect of granular cell-conditioned medium could be blocked by the addition of the cell adhesion recognition sequence, RGDS, but not by RGES. These results demonstrate experimentally that granular cells are required for encapsulation of foreign targets by plasmatocytes in vitro, and that the role of granular cells in this process involves an RGD-dependent cell adhesion mechanism.


The DiGeorge syndrome and the fetal alcohol syndrome.
Four patients with clinical and laboratory features of the DiGeorge syndrome had a definite history of maternal alcoholism. Certain clinical abnormalities in the patients were characteristic of the DiGeorge syndrome as well as the fetal alcohol syndrome, including abnormalities of the eyes, ears, mouth, face, cardiovascular system, CNS, and immune system. Maternal alcoholism may result in the DiGeorge syndrome either as the single causative factor or in conjunction with other as yet unknown teratogens.


Laser capture microdissection for the analysis of gene expression during embryogenesis of Arabidopsis.
It is during embryogenesis that the body plan of the developing plant is established. Analysis of gene expression during embryogenesis has been limited due to the technical difficulty of accessing the developing embryo. Here we demonstrate that laser capture microdissection can be applied to the analysis of embryogenesis. We show how this technique can be used in concert with DNA microarray for the large-scale analysis of gene expression in apical and basal domains of the globular-stage and heart-stage embryo, respectively, when critical events of polarity, symmetry and biochemical differentiation are established. This high resolution spatial analysis shows that up to approximately 65% of the genome is expressed in the developing embryo, and that differential expression of a number of gene classes can be detected. We discuss the validity of this approach for the functional analysis of both published and previously uncharacterized essential genes.


Bupivacaine anesthesia in retinal detachment surgery.
Retinal detachment surgery and other long-duration ocular procedures can be performed safely with local anesthesia, but a long-acting anesthetic is required. A randomized and masked prospective series of 126 scleral buckling procedures was undertaken to test a mixture of bupivacaine (Marcaine), lidocaine (Xylocaine), and epinephrine against lidocaine and epinephrine. The data collected show a conclusive superiority of the bupivacaine mixture in longevity of action and overall performance. Also, there was significantly less variability in the quality of anesthesia with the mixture than with lidocaine (p = .0015).


The Role of Working Memory in Early Literacy and Numeracy Skills in Kindergarten and First Grade.
The working memory system supports learning processes such as acquiring new information and the development of new skills. Working memory has been found to be related to both early literacy and early numeracy in kindergarten and to linguistic and mathematical academic skills at older ages, but the contribution of each of the memory components at these ages is not yet clear. The purpose of this study is to examine the unique connections among the various systems of WM, early literacy, and early numeracy using various assessment tests of simple WM and complex WM, as well as a variety of tasks in math and language skills administered to the same 250 children in kindergarten and 150 children in first grade. Consistent with the predictions, significant relations among all components of memory and mathematics and language knowledge at both ages were found, although these connections were differential for the different types of tasks and memory systems. The connection of complex WM was stronger in its contribution and more significant in first grade in both mathematics and language domains. Complex WM resources were more important in early literacy at kindergarten age, while simple WM seems to be important in early numeracy. The theoretical and educational implications of these results are discussed accordingly.


Staining proteins in gels with silver nitrate.
INTRODUCTIONSilver staining is one of the commonly used procedures for visualizing proteins in acrylamide gels. All silver staining methods rely on the reduction of ionic to metallic silver to provide metallic silver images; the selective reduction at gel sites occupied by proteins compared to nonprotein sites is dependent on differences in the oxidation-reduction potentials at these sites. There are two broad methodologies for silver staining. One approach (nondiamine silver nitrate stains) uses silver nitrate as the silvering agent and formaldehyde in alkaline carbonate solution as the developing agent, whereas the other approach (diamine or ammoniacal stains) uses ammoniacal silver as the silvering agent and formaldehyde in dilute citric acid as the developing agent. Although protocols using ammoniacal silver are arguably more sensitive and give darker hues than those based on silver nitrate, they are more prone to negative staining, resulting in hollow or "doughnut" spots, give unacceptable backgrounds with tricine-based gel systems, and are not very robust because of their reliance on the ammonia-silver ratio. Additionally, ammoniacal silver staining is more sensitive for basic proteins but less so for very acidic proteins. This protocol describes a silver nitrate staining approach. Its sensitivity is in the low-nanogram range, which is 50-100 times more sensitive than classical Coomassie Blue staining, ~10 times better than colloidal Coomassie Blue staining, and at least twice as sensitive as the zinc/imidazole negative staining method.


Levomilnacipran for the treatment of major depressive disorder: a review.
Levomilnacipran (LVM, Fetzima(®)) was recently approved by the US Food and Drug Administration for the treatment of major depressive disorder. It is a unique dual neurotransmitter reuptake inhibitor. In contrast with other selective serotonin norepinephrine reuptake inhibitors, including duloxetine, venlafaxine, and desvenlafaxine, it has greater selectivity for inhibiting norepinephrine reuptake than serotonin reuptake. Our review focuses on the efficacy, safety, and tolerability data for five double-blind, placebo-controlled, short-term studies and two long-term studies. In the short-term studies, LVM was found to be more effective than placebo in reducing depression (Montgomery-Åsberg Depression Rating Scale) scores as well as improving functional impairment (Sheehan Disability Scale) scores. Long-term studies found LVM to be similarly effective but in the only placebo-controlled long-term study, LVM was not significantly superior to placebo. LVM is fairly well tolerated, with the most common adverse events being nausea, headache, dry mouth, hyperhidrosis, and constipation. Discontinuation rates were mildly increased in those being treated with LVM (9%) versus placebo (3%). Adverse events were not dose-related except for urinary hesitancy and erectile dysfunction. LVM was weight neutral, was not toxic to the liver, and did not cause clinically significant QTc prolongation. Consistent with being a predominant potentiator of norepinephrine, pulse and blood pressure were significantly elevated by LVM but rarely induced tachycardia or hypertension. LVM is a relatively safe alternative antidepressant treatment with minimal drug-drug interactions. It is the only antidepressant that has in its labeling that it is not only effective in improving depression but also effective in improving impaired functioning. Whether this important effect on functioning is unique to LVM must be researched. In addition, whether LVM might be effective in norepinephrine-deficit depression, refractory depression, atypical depression, or seasonal depression is yet to be evaluated. Ultimately, head-to-head studies comparing LVM with other antidepressants will determine the place of LM in antidepressant treatment.


Iron chelator deferoxamine alters iron-regulatory genes and proteins and suppresses osteoblast phenotype in fetal rat calvaria cells.
There are few studies describing the extent to which low iron status affects osteoblastogenesis, despite evidence that iron deficiency produces adverse effects on bone density. The purpose of this study was to evaluate alterations in intracellular iron status by measuring iron-regulated gene and protein expression and to describe development of osteoblast phenotype in primary cells treated with iron chelator deferoxamine (DFOM) during differentiation. Using the well-described fetal rat calvaria model, cells were incubated with 0-8 microM DFOM throughout differentiation (confluence to day (D) 21), or only during early differentiation (confluence to D13-15) or late differentiation (D13-15 to D21). Changes in intracellular iron status were determined by measuring alterations in gene and protein expression of transferrin receptor and ferritin light chain and heavy chain. Development of osteoblast phenotype was monitored by measuring expression of genes that are known to be up-regulated during differentiation, analyzing the percentage of mineralized surface area, and counting the number of multi-layered bone nodules at the end of culture. Results indicate that treatment throughout differentiation with 8 microM DFOM alters iron-regulated genes and proteins by mid-differentiation (D13-15) in a pattern consistent with iron deficiency with concomitant down-regulation of osteoblast phenotype genes, especially osteocalcin. Additionally, alkaline phosphatase staining was lower and there was about 70% less mineralized surface area (p<0.05) by D21 in wells treated throughout differentiation with 8 microM DFOM compared to control. Down-regulation of osteocalcin and alkaline phosphatase mRNA (p<0.05) and suppressed mineralization (p<0.05) was also evident at D21 in cells treated only during early differentiation. In contrast, treatment during late differentiation did not alter osteoblastic outcomes by D21. In conclusion, it appears that iron is required for normal osteoblast phenotype development, and that early rather than late differentiation events may be more sensitive to iron availability.


Chronic recurrent multifocal osteomyelitis: a case report and review of literature.
Chronic recurrent multifocal osteomyelitis (CRMO) is a rare condition of largely unknown aetiology and pathogenesis with variable clinical and radiological features. There are no reports on CRMO to the best of our knowledge; in the West African sub region. To present a case and review the literature on chronic recurrent multifocal osteomyelitis. A 13-year-old male presented with a three-year history of recurrent discharging right thigh sinus and an 11-day history of a left shoulder swelling with discharging sinus. Detailed history was obtained and physical examination and radiological and microbiological tests carried out before treatments. There were periodic exacerbations of pain, swelling and discharge over affected areas. He had a short limb gait and shoulder asymmetry. His left shoulder was warm, erythematous and there was decreased range of movement in all directions. Investigations revealed an erythrycyte sedimentation rate (ESR) of 150mm/hr. Wound swabs taken at different times from the right thigh and shoulder sinuses revealed no growth. Radiographs of the left arm, right thigh and hip showed features consistent with chronic osteomyelitis. There were associated destruction of the left hip and soft tissues welling in the left shoulder. He was principally treated with non-steroidal anti-inflammatory drugs (NSAIDs) , antibiotics and dressing of sinuses for three months and had only mild relief of clinical features but no improvement in radiological picture. Chronic recurrent multifocal osteomyelitis is a diagnosis of exclusion which is usually under diagnosed because amongst other things, not much is known about it. Successful treatment is difficult to achieve, though some authors have reported good results with combined medical and surgical treatment.


Kaempferol Improves Breast Cancer-Related Depression through the COX-2/PGE2 Pathway.
Breast cancer-related depression (BCRD) is strongly associated with BC and increases recurrence and mortality. This study investigated the role of kaempferol in the pathogenesis of BCRD and its underlying mechanism. 4T1 mouse BC cells were treated with corticosterone (Cort) in vitro to develop a neuronal injury model, and a BCRD mouse model was established by injecting 4T1 cells and Cort. The effects of kaempferol on 4T1 cells and BCRD models were measured by behavioral tests, Cell Counting Kit-8 assay, wound healing assay, colony formation assay, Western blot analysis, quantitative real-time PCR, hematoxylin and eosin staining, enzyme-linked immunosorbent assay, and immunofluorescence. BCRD cells were transfected with the cyclo-oxygenase-2 (COX-2) overexpression plasmid to study the role of the COX-2/prostaglandin E2 (PGE2) axis in the anti-BCRD activity of kaempferol. The connection between kaempferol and COX-2 was analyzed by molecular docking. Kaempferol reduced the viability, migration, and clones of 4T1 cells and inhibited BC growth and depression-like behavior in mice. Kaempferol alleviated inflammation in BCRD, decreased interleukin 1 beta (IL-1β) and IL-6 levels, and increased transforming growth factor beta 1 (TGF-β1) and IL-10 levels. In addition, kaempferol elevated the levels of serotonin, dopamine, and norepinephrine and the amount of 5-Bromo-2'-deoxyuridine/neuronal nuclei-positive cells. Kaempferol downregulated COX-2 and PGE2, and kaempferol could dock with the protein structure of COX-2. Overexpression of COX-2 reduced BCRD viability, upregulated IL-1β and IL-6 levels, and downregulated TGF-β1 and IL-10 expression. Overexpression of COX-2 reversed the protective effects of kaempferol. Kaempferol exerted anti-BCRD effects, at least in part by inhibiting the COX-2/PGE2 pathway, which regulates neuroinflammation, neurotransmitter imbalance, and defective neurogenesis. Therefore, kaempferol may be a promising candidate active ingredient for treating BCRD.


Effects and experiences of warm versus cold skin disinfection.
The intention of preoperative skin antisepsis is to reduce the risk of postoperative surgical wound infections through the prevention of contamination of the wound site from the patient's skin flora, and to provide a sterile work area. This pilot study aimed to compare warm versus cold skin disinfection with a chlorhexidine solution on bacterial growth, skin temperatures and the experiences of the disinfection. Ten healthy volunteers without any association to Sweden's healthcare services were included in an experimental study with crossover design. The results showed that warm (38°C) disinfection seems to be comparable with cold (20°C) disinfection in reducing bacterial growth. Experiences differ, but warm disinfection is generally considered more pleasant than cold as a result of heat-loss to the skin. However, further research with larger sample sizes and on patients undergoing surgery is needed.


Cell-type-specific expression of epidermal cytokeratin genes during gastrulation of Xenopus laevis.
Analysis of the spatial pattern of expression of embryo-specific epidermal cytokeratin genes in Xenopus laevis shows earliest activity in the animal pole cells of stage-9 blastulae. These genes are transcribed predominantly in the epithelial or outer ectoderm, to a lesser extent in the sensorial or inner ectoderm, and at low levels if at all in other regions of the embryo. In the early gastrula the entire ectoderm, including preneural and preepidermal regions, expresses cytokeratin mRNAs; accumulation of these mRNAs in preneural cells is terminated after contact is made with involuting chordamesoderm. On the basis of this and earlier work (Sargent et al. 1986) we suggest that the pattern of expression of cytokeratin genes in frog embryogenesis is based on prelocalized components modulated by the inductive influence of involuting chordamesoderm. The cytokeratin proteins are deposited in the form of filamentous networks in both layers of the epidermis. In the epithelial layer, a much denser mesh of filaments is facing the outside of the embryo. This polarity is established at the onset of the polymerization of these filaments. Thus, intraembryonic and intracellular localization of keratin gene expression and protein deposition is established at the onset of activation of these genes.


Attitudes of paediatricians to HIV and hepatitis B virus infection.
There is a recent upsurge of interest among health care professionals regarding the risk of accidental occupational exposure to HIV virus. We evaluated knowledge of virus carriage prevalence, needlestick injuries, venepuncture practices, and glove use among paediatricians in Wales and South West Regional Health Authorities. We also attempted to evaluate hepatitis B immunisation uptake in this group. Paediatricians have traditionally been considered a low risk group in the context of accidental occupational exposure to these viruses. We targeted a four point questionnaire at 221 paediatricians. Results suggested that despite recent increasing concern about these viruses, that is reflected in the amount of medical literature recently published, and the issuing of Department of Health guidelines on venepuncture, knowledge of prevalence of HIV and hepatitis B carriage rates, and hence assessment of risk magnitude, was surprisingly poor. Safe venepuncture practices were not widely used. In the 12 months before receiving the questionnaire 55% had suffered a needlestick injury with only 10% reporting the fact. Hepatitis B immunisation uptake was highest in the junior grades (but this does not necessarily mean those at greatest risk). There were many inconsistencies between the clinicians' perceptions of risk and their practices. As the virus attains a firm hold in the heterosexual population paediatricians by virtue of the nature of venepuncture in children will almost certainly see their risk of acquiring HIV/hepatis B viruses secondary to accidental occupational exposure increase over the next decade. Without an improvement in current knowledge of carriage prevalence in high risk areas and alteration in venepuncture practices/hepatitis B immunisation uptake some will unfortunately, though avoidably, contract these bloodborne viral infections.


Genetic analysis of type 1 diabetes: embryonic stem cells as new tools to unlock biological mechanisms in type 1 diabetes.
The nonobese diabetic (NOD) mouse has provided an important animal model for studying the mechanism and genetics of type 1 diabetes over the past 30 years. Arguably, the bio-breeding (BB) rat model may be an even closer phenotypic mimic of the typical human disease. A large number of distinct genetic traits which influence diabetes development have been defined through an extraordinary effort, most conspicuously in the mouse model. However, in both NOD and BB models the lack of availability of robust means for experimental genetic manipulation has restricted our understanding of the mechanisms underlying this spontaneous autoimmune disease. Recent developments in the derivation of embryonic stem (ES) cells have the potential to transform this picture. We argue here that targeting of NOD strain ES cells can bring much needed certainty to our present understanding of the genetics of type 1 diabetes in the NOD mouse. In addition, ES cells can play important roles in the future, in both the NOD mouse and BB rat models, through the generation of new tools to investigate the mechanisms by which genetic variation acts to promote diabetes.


Pharmacological characterization of two calcium currents in GH3 cells.
Whole cell patch-clamp techniques were used to investigate the pharmacological properties of calcium currents in the clonal rat pituitary cell line GH3. Current traces induced by a 100-ms pulse to 0 mV from a holding potential of -80 mV consisted of a component that rapidly inactivated during the pulse and a component that slowly inactivated during the pulse. When the holding potential was reduced to -32 mV, the rapidly inactivating component of the trace disappeared. The dihydropyridine calcium channel blocker nitrendipine affected only the slowly inactivating component of the trace. At a holding potential of -80 mV, nitrendipine blocked the slowly inactivating current with an IC50 of 1 microM. The IC50 for nitrendipine was found to be dependent on the holding potential, decreasing to 10 nM when the holding potential was -32 mV. The dihydropyridine agonist Bay-K 8644, like nitrendipine, affected only the slowly inactivating component. The inorganic blocker Cd2+ blocked both components but the slowly inactivating current was three- to fourfold more sensitive. These results are best explained by the existence of two types of calcium channels in these cells, one sensitive to dihydropyridines and one insensitive to dihydropyridines. These channels appear analogous to the T-type channel (inactivating current) and L-type channel (slowly inactivating current) described in other preparations.


Drug Resistance Mutations Against Protease, Reverse Transcriptase and Integrase Inhibitors in People Living With HIV-1 Receiving Boosted Protease Inhibitors in South Africa.
The South African national combination antiretroviral therapy (cART) roll-out program started in 2006, with over 4.4 million people accessing treatment since it was first introduced. HIV-1 drug resistance can hamper the success of cART. This study determined the patterns of HIV-1 drug-resistance associated mutations (RAMs) in People Living with HIV-1 (PLHIV-1). Receiving first (for children below 3 years of age) and second-line (for adults) cART regimens in South Africa. During 2017 and 2018, 110 patients plasma samples were selected, 96 samples including those of 17 children and infants were successfully analyzed. All patients were receiving a boosted protease inhibitor (bPI) as part of their cART regimen. The viral sequences were analyzed for RAMs through genotypic resistance testing. We performed genotypic resistance testing (GRT) for Protease inhibitors (PIs), Reverse transcriptase inhibitors (RTIs) and Integrase strand transfer inhibitors (InSTIs). Viral sequences were subtyped using REGAv3 and COMET. Based on the PR/RT sequences, HIV-1 subtypes were classified as 95 (99%) HIV-1 subtype C (HIV-1C) while one sample as 02_AG. Integrase sequencing was successful for 89 sequences, and all the sequences were classified as HIV-1C (99%, 88/89) except one sequence classified CRF02_AG, as observed in PR/RT. Of the 96 PR/RT sequences analyzed, M184V/I (52/96; 54%) had the most frequent RAM nucleoside reverse transcriptase inhibitor (NRTI). The most frequent non-nucleoside reverse transcriptase inhibitor (NNRTI) RAM was K103N/S (40/96, 42%). Protease inhibitor (PI) RAMs M46I and V82A were present in 12 (13%) of the sequences analyzed. Among the InSTI major RAM two (2.2%) sequences have Y143R and T97A mutations while one sample had T66I. The accessory RAM E157Q was identified in two (2.2%). The data indicates that the majority of the patients failed on bPIs didn't have any mutation; therefore adherence could be major issue in these groups of individuals. We propose continued viral load monitoring for better management of infected PLHIV.


Preliminary evidence of sex differences in behavioral and neural responses to palatable food reward in rats.
The female bias in eating disorder prevalence is the largest in all of psychiatry. Binge eating on palatable food (PF) is a core, maladaptive symptom that cuts across all major types of eating disorders and can be studied via animal models. Using an individual differences rat model of binge eating that identifies binge eating prone (BEP) and binge eating resistant (BER) phenotypes, we previously showed that, compared with males, females consume more PF and are more likely to be classified as BEP. One potential explanation for this sex difference is that PF is inherently more rewarding to females, leading to higher rates of binge eating. Here we tested the hypothesis that females have more robust behavioral and neural responses to PF reward than males. Adult male (N=18) and female (N=17) Sprague-Dawley rats were exposed to the Conditioned Place Preference paradigm using PF as the unconditioned stimulus. Select males (N=9) and females (N=9) were video-recorded during three of the PF-paired conditioning sessions to score feeding behavior. Following CPP, 13 male and 12 female rats were exposed to PF just prior to sacrifice to induce expression of the neural activation marker Fos, and Fos expression was quantified in mesocorticolimbic, hypothalamic, and amygdalar circuits. In the CPP paradigm, females displayed a more robust shift in preference for the chamber paired with PF compared with males, and behavioral analyses revealed that average duration of individual feeding bouts during pairing sessions was longer in females than in males. Fos expression was significantly higher in females vs. males in select regions of the mesocorticolimbic reward circuit, with no sex differences in hypothalamic or amygdalar regions. These results provide initial evidence that PF may be more rewarding to females than to males, possibly due to heightened responsiveness of neural substrates that mediate the hedonic and motivational responses to PF, which in part, may underlie sex differences in binge eating proneness.


Hepatocellular injury induced by chronic low-dose CCl4 in spontaneous and renal hypertensive rats: a correlation to the reversal of experimental rat hypertensive models.
Renal hypertension has been reduced to normal levels in the rat with subcutaneous injections of carbon tetrachloride (CCl4). Previous light microscopic evaluation of liver sections demonstrated varying degrees of hepatocellular and hepatic parenchymal injury secondary to CCl4 administration. In normotensive rat populations, saline injections have been shown not to cause hypertension, and the treatment of normal rats with subcutaneous CCl4 did not change their blood pressure over that of noninjected controls. In this study we compare the extent of blood pressure reduction with the degree of CCl4-induced liver injury in the renal-induced, spontaneous hypertensive rat (SHR), and normotensive rat. Statistically significant blood pressure reduction followed CCl4 injection of renal (p less than .01) and SHR (p less than .001) hypertensive animals. The morphological liver injury appeared most sensitively reflected in hepatocellular nuclear atypia. The SHR animals were the most resistant to CCl4 liver damage by chronic CCl4 treatment. There was no measurable effect of CCl4 treatment on the animals as assessed by their activity, rate of development and weight gain. The degree of blood pressure reduction due to CCl4 treatment in the SHR animals was similar to that previously reported in DOCA and renal-induced hypertensive animals.


Minimally Invasive Therapy for Upper Tract Urothelial Cell Cancer.
Minimally invasive therapy for upper tract urothelial cell cancer has assumed an important role in the management of this relatively uncommon genitourinary malignancy. In this review, current standards for patient selection, surgical approach, as well as overall outcomes will be discussed. Moreover, the use of intraluminal agents in upper tract disease is examined along with its efficacy and associated complications.


Perioperative Outcomes of Radical Trachelectomy in Early-Stage Cervical Cancer: Vaginal Versus Laparoscopic Approaches.
This study aimed to compare the laparoscopic-assisted radical vaginal trachelectomy (LARVT) and laparoscopic radical trachelectomy (LRT) surgical approaches and provide outcome data on patients who have undergone radical trachelectomy. We identified patients who had undergone LARVT or LRT at Samsung Medical Center between January 2005 and March 2013. A total of 38 patients were identified, and 21 patients had undergone LARVT, whereas 17 patients had undergone LRT. The median age was 32 years for both groups. Most of the patients had a squamous cell carcinoma (68.4%) and International Federation of Gynecology and Obstetrics stage IB1 disease (76.3%). Twenty (52.6%) of 38 patients had tumor size greater than 2 cm. There were no significant differences between groups in the baseline characteristics except for the tumor size. Patients undergoing LRT had significantly larger tumor size than patients undergoing LARVT (median tumor size, 2.7 cm [range, 1.2-3.7] vs 2.1 cm [range, 0.4-3.0], P = 0.032). Perioperative outcomes were similar between groups except for the decline of hemoglobin after surgery. The median decline of hemoglobin indicating blood loss was significantly smaller in the LRT group than in the LARVT group (1.8 g/dL [range, 0.5-3.5] vs 2.6 g/dL [range, 0.7-6.2], P = 0.017). Intraoperative complications occurred in 2 patients (9.5%, 2/21) in LARVT group. Although 52.6% of tumors were larger than 2 cm, recurrence occurred only in 3 (7.9%) patients who underwent LARVT. The study shows the feasibility of LRT, with the advantage of reduced blood loss. The LRT could be an alternative option for patients with large tumors. Further researches are needed to investigate the long-term outcomes.


Predicting antigenic variants of H1N1 influenza virus based on epidemics and pandemics using a stacking model.
H1N1 is the earliest emerging subtype of influenza A viruses with available genomic sequences, has caused several pandemics and seasonal epidemics, resulting in millions of deaths and enormous economic losses. Timely determination of new antigenic variants is crucial for the vaccine selection and flu prevention. In this study, we chronologically divided the H1N1 strains into several periods in terms of the epidemics and pandemics. Computational models have been constructed to predict antigenic variants based on epidemic and pandemic periods. By sequence analysis, we demonstrated the diverse mutation patterns of HA1 protein on different periods and that an individual model built upon each period can not represent the variations of H1N1 virus. A stacking model was established for the prediction of antigenic variants, combining all the variation patterns across periods, which would help assess a new influenza strain's antigenicity. Three different feature extraction methods, i.e. residue-based, regional band-based and epitope region-based, were applied on the stacking model to verify its feasibility and robustness. The results showed the capability of determining antigenic variants prediction with accuracy as high as 0.908 which performed better than any of the single models. The prediction performance using the stacking model indicates clear distinctions of mutation patterns and antigenicity between epidemic and pandemic strains. It would also facilitate rapid determination of antigenic variants and influenza surveillance.


A pragmatic parallel arm randomized-controlled trial of a multi-pronged electronic health record-based clinical decision support tool protocol to reduce low-value antipsychotic prescriptions among older adults with Alzheimer's and related dementias.
Among patients with Alzheimer's disease and its related dementias (ADRD) with behavioral disturbances, antipsychotic prescriptions have limited efficacy and increase the risk of death. Yet, physicians continue to routinely prescribe low-value antipsychotic medications for behavioral disturbances among patients with ADRD. We designed a pragmatic randomized-controlled trial to measure the impact of a behavioral economic electronic health record (EHR) clinical decision support (CDS) intervention to reduce physician prescriptions of new antipsychotic medications among patients with ADRD. Utilizing a pragmatic parallel arm randomized-controlled trial design, the study will randomize eligible physicians from a large academic health system to either receive a EHR CDS intervention or not (control) when they prescribe a new antipsychotic medication during visits with patients with ADRD. The intervention will include three components: 1) alerts prescribers that antipsychotic prescriptions increase mortality risk (motivating physicians' intrinsic desire for non-malfeasance); 2) offers non-pharmacological behavioral resources for caregivers; 3) auto-defaults the prescription to contain the lowest dose and number of pill-days (n = 30) without refills if the prescriber does not cancel the order (appealing to default bias). Over 1 year, we will compare the cumulative total of new antipsychotic pill-days prescribed (primary outcome) by physicians in the intervention group versus in the control group. The study protocol meets international SPIRIT guidelines. Behavioral economics, or the study of human behavior as a function of more than rational incentives, considering a whole host of cognitive and social psychological preferences, tendencies, and biases, is increasingly recognized as an important conceptual framework to improve physician behavior. This pragmatic trial is among the first to combine two distinct behavioral economic principles, a desire for non-malfeasance and default bias, to improve physician prescribing patterns for patients with ADRD. We anticipate this trial will substantially advance understanding of how behavioral-economic informed EHR CDS tools can potentially reduce harmful, low-value care among patients with ADRD.


Effects of fuels, engine load and exhaust after-treatment on diesel engine SVOC emissions and development of SVOC profiles for receptor modeling.
Diesel exhaust emissions contain numerous semivolatile organic compounds (SVOCs) for which emission information is limited, especially for idling conditions, new fuels and the new after-treatment systems. This study investigates exhaust emissions of particulate matter (PM), polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs (NPAHs), and sterane and hopane petroleum biomarkers from a heavy-duty (6.4 L) diesel engine at various loads (idle, 600 and 900 kPa BMEP), with three types of fuel (ultra-low sulfur diesel or ULSD, Swedish low aromatic diesel, and neat soybean biodiesel), and with and without a diesel oxidation catalyst (DOC) and diesel particulate filter (DPF). Swedish diesel and biodiesel reduced emissions of PM2.5, Σ15PAHs, Σ11NPAHs, Σ5Hopanes and Σ6Steranes, and biodiesel resulted in the larger reductions. However, idling emissions increased for benzo[k]fluoranthene (Swedish diesel), 5-nitroacenaphthene (biodiesel) and PM2.5 (biodiesel), a significant result given the attention to exposures from idling vehicles and the toxicity of high-molecular-weight PAHs and NPAHs. The DOC + DPF combination reduced PM2.5 and SVOC emissions during DPF loading (>99% reduction) and DPF regeneration (83-99%). The toxicity of diesel exhaust, in terms of the estimated carcinogenic risk, was greatly reduced using Swedish diesel, biodiesel fuels and the DOC + DPF. PAH profiles showed high abundances of three and four ring compounds as well as naphthalene; NPAH profiles were dominated by nitro-naphthalenes, 1-nitropyrene and 9-nitroanthracene. Both the emission rate and the composition of diesel exhaust depended strongly on fuel type, engine load and after-treatment system. The emissions data and chemical profiles presented are relevant to the development of emission inventories and exposure and risk assessments.


Immunohistochemical demonstration of metallothionein in eyes with choroidal melanomas.
The since immunohistochemically detectable metallothionein (MT) overexpression has been described in a variety of human tumours, including skin melanomas, in relation to different stages of tumour development and progression. We used a monoclonal antibody to investigate the distribution of MT in 18 formalin-fixed, paraffin-embedded, surgically enucleated eyes with choroidal melanomas, from 18 patients (8 male, 10 female; age range 30-83 years, mean 58.7). Clinico-pathological details and follow-up data (2-124 months, mean 36.1) were also available. MT immunoreactivity was recorded and the percentage of stained cells was graded for semiquantitative purposes. Correlations between immunohistochemical data and morphological characteristics of melanomas were investigated using non-parametric methods; survival analysis was done by the Kaplan-Meier method and the survival curves were compared by the Mantel-Cox log-rank test. MT immunoexpression was found in 15/18 cases (83.3%) with staining scores from 1 to 3; MT staining varied in intensity and was mainly localized in the cytoplasm, although a combined nuclear/cytoplasmic reactive pattern was seen in neoplastic elements. No differences in MT immunostaining were seen in relation to age or sex, tumour size, histotype and amount of pigment; univariate analysis of survival data showed no prognostic significance regarding MT expression. The immunohistochemical evidence of MT in neoplastic elements could be related to the production of this scavenging protein in the tumour for cell defense mechanisms against hydroxyl free radicals, and to act as a Zn donor, since Zn is required for the synthesis of DNA and DNA-repair enzymes.


Identification of an attenuating region in the bovine follicle-stimulating hormone beta subunit mRNA that decreases its expression in E. coli.
Follicle-stimulating hormone (FSH) is one of the key regulators of gonadal function in mammals. Recombinant DNA expression of this hormone has proved to be a difficult task as expression levels are invariably low, irrespective of the expression system employed. In the present study, we have attempted to identify reasons for this low expression using bacterial expression vectors, and we report here the identification of a theoretically predicted hairpin structure in the mRNA corresponding to the N-terminal portion of the mature coding portion of bFSHbeta cDNA that is responsible for attenuating its expression in E. coli. When full-length FSHbeta was expressed using the bacterial expression vector, a very low expression was obtained. However, when fragments of FSHbeta with N-terminal deletions (amino acids 24-110 and 13-110) were expressed using the same expression strategy, a 30- to 40-fold higher expression was observed. This low expression of FSHbeta could be attributed to a hairpin structure present in the first 12 codons of mature FSHbeta mRNA. Disruption of this structure without changing the amino acid sequence resulted in a higher level of expression of FSHbeta. The predicted hairpin structure, though away from the transcriptional and translational start site, was able to downregulate the expression of FSHbeta probably by impeding the movement of ribosomes.


PLANETARY SCIENCE: 'Spiders' Channel Mars Polar Ice Cap.
Scientists studying the latest high-resolution photos of the martian south polar ice cap think they may have found additional clues to its ebb and flow. These hints of the planet's bizarre atmosphere come from a new class of dramatic-looking terrain features whose dark, multilimbed, vaguely radial designs have earned them the moniker "black spiders," and another group of dusky, spreading features called "dark fans." At a recent gathering here of Mars researchers, a planetary scientist proposed that the spiders might be subsurface gas channels, visible through an unusually transparent section of the martian ice.


Current treatment strategies for clozapine-induced sialorrhea.
To provide an understanding of the underlying pathophysiology and current treatment options for clozapine-induced sialorrhea. Literature was retrieved through MEDLINE (1977-February 2011) using the key search terms clozapine, sialorrhea, hypersalivation, drooling, and treatment. In addition, reference citations from identified publications were reviewed. All articles published in English identified from the data source were evaluated and included in the review. Sialorrhea is a common and disabling adverse effect of clozapine use. Current treatment options include topical and oral antimuscarinic medications and α-adrenergic agents. New areas of investigation include glycopyrrolate, botulinum toxin, and substitute benzamide derivatives. Thirteen clinical trials (2 retrospective, 5 open-label, 6 double-blind) and 13 case reports were reviewed. Overall, there are weak data on use of antimuscarinic agents, consisting mostly of small open-label or retrospective studies. Glycopyrrolate, however, demonstrated significant reduction of hypersalivation in a randomized controlled trial. Medications with activity at α-adrenergic receptors have shown positive results in case reports, retrospective evaluations, and an open-label trial, but have not been investigated in a double-blind, controlled fashion. Botulinum toxin also significantly improved sialorrhea in both a case report and double-blind study, although the trial included hypersalivation from other etiologies in addition to clozapine. Substitute benzamide derivatives have demonstrated significant improvements in randomized controlled trials; however, they are not available in the US. Overall, few treatment strategies have been evaluated in controlled settings, warranting further randomized controlled trials to identify more effective treatment options. Current pharmacologic treatment options for clozapine-induced sialorrhea are limited in number and efficacy. Although few randomized controlled trials have been conducted, this review identifies potential treatment alternatives for this common and sometimes severe adverse effect.


[Transorbital penetrating injury without sequlae].
A 33-year-old man was admitted to hospital with a butter knife stuck in his head after having attempted to commit suicide by forcing the knife through his skull. Physical examination revealed a stable patient with only minimal associated haemorrhage. A CT scan revealed, that the penetrating knife had passed through the sphenoid bone, the posterior orbit and the ethmoid cells and ended in the contralateral nasal cavity. The knife was removed by its own trajectory. After surgery, the patient was neurologically intact with normal sight and well-functioning eye movements, and he made a full recovery.


Mass media as a population-level intervention tool for Chlamydia trachomatis screening: report of a pilot study.
To determine the feasibility and affect of mass media use in a population-level intervention for chlamydia screening promotion. A population-level chlamydia intervention protocol was field tested. The intervention, targeting 15-25-year-old individuals, was designed to: (a) increase awareness of personal risk for chlamydial infection; (b) facilitate dissemination of chlamydia knowledge by use of a telephone hot line; and (c) promote care-seeking behavior (report for a chlamydia screening program). The intervention activities included: (a) mail outreach, (b) a television and radio campaign, (c) a prerecorded Check-It-Out chlamydia hot line, (d) a staffed chlamydia Options information line, and (e) a free confidential urine ligase chain reaction (LCR) test for chlamydia. Mass mailings were scheduled at intervals, starting two-weeks before the beginning of the television advertisement. The 30-second television advertisement was aired on local television stations 130 times in a 6-week period. The outcome measures were quantity and characteristics of incoming calls to the automated hot line and staffed chlamydia information phone line in response to the chlamydia campaign, and response to the urine screening program. Descriptive and bivariate analyses were used to evaluate the outcomes. The hot line was called 642 times during the monitoring period (November 1, 1999 to March 8, 2000), the majority (92%) during the 6 weeks of television advertisement, with an average of 99 calls per week, compared with an average of 9 calls per week after the commercial ended. Each bulk mailing was accompanied by a boost in the incoming hot line calls. The research staff triaged 133 calls to the "Options" phone line, 81% in the 6 weeks of the TV ad. The mean age of the 133 callers was 23.9 +/- 7.7 years (range 14-49 years). A majority called for screening information; 67% of callers were females and 84% of female callers were under age 26 years. Five percent of callers identified themselves as a parent of a teenager. The majority credited the TV ad as their source of the hot line number. Thirty-one individuals reported for a confidential chlamydia screening, 27 of 31 (87%) during the 6 weeks of TV advertising. No negative responses regarding the chlamydia campaign were encountered. This report describes strategies used to implement and measure the effectiveness of a mass media campaign and demonstrates evidence that mass media is effective in delivering STD intervention messages to young people.


Continuous monitoring of enzyme reactions on a microchip: application to catalytic RNA self-cleavage.
Kinetic analysis of RNA enzymes, or ribozymes, typically involves the tedious process of collecting and quenching reaction time points and then fractionating by polyacrylamide gel electrophoresis (PAGE). As a way to automate and simplify this process, continuous analysis of a ribozyme reaction is demonstrated here using completely automated capillary sample introduction onto a microfabricated device with laser-induced fluorescence detection. The method of injection is extremely reproducible thereby standardizing data analysis. A 30-nucleotide ribozyme model, the self-cleaving lead-dependent ribozyme, or "leadzyme", which cleaves into a 24-mer and a 6-mer in the presence of Pb(2+), was end-labeled with fluorescein (FAM) and used to demonstrate the potential of this technique. After manually initiating the cleavage reaction by Pb(2+) addition, reaction samples were automatically injected directly into the parallel separation lanes of the chip via a capillary at predetermined time intervals, thus eliminating the need for additional sample-handling steps. The FAM-labeled leadzyme starting material and products were monitored for 60 min in order to ascertain kinetic information. The effect of lead acetate concentration on cleavage rates was also studied, and the results are in agreement with rates determined by conventional hand-mixing/PAGE analysis. This work demonstrates, through the use of a simple ribozyme model, the potential of this method to provide valuable kinetic information for other, more complex, biologically relevant RNA and protein enzymes.


Afferent renal inputs onto subfornical organ neurons responsive to angiotensin II.
Experiments were done in pentobarbital sodium-anesthetized rats to investigate the effect of electrical stimulation of afferent renal nerves (ARN) on the discharge rate of subfornical organ (SFO) neurons that responded to changes in plasma levels of angiotensin II (ANG II) and projected directly to the paraventricular nucleus of the hypothalamus (PVH). Extracellular recordings were made from 76 histologically verified single neurons in the SFO that were excited by intracarotid infusions of ANG II. Of these units, 54.8% (23 of 42) responded with excitation to ARN stimulation (mean onset latency, 125 +/- 35 ms). None of the SFO units excited by plasma ANG II were found to be inhibited by ARN stimulation. An additional 34 units in the SFO that were excited by plasma ANG II were also antidromically activated by stimulation of the PVH. Of these neurons, 17.8% (6 of 34) were also excited by stimulation of ARN. The results indicate that inputs from ARN converge onto SFO neurons that alter their discharge rate during changes in plasma concentration of ANG II and project directly to the PVH. These data suggest that ARN may play an important role in body fluid balance and circulatory regulation by modulating the activity of SFO neurons that function in the detection of blood-borne signals resulting from the decrease in extracellular fluid volume and arterial pressure and that influence the activity of hypothalamic nuclei that contain neurosecretory neurons.


Impact of developmental maturity of soybean on the seasonal abundance of soybean aphid (Hemiptera: Aphididae).
The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), alternates between a primary overwintering host (buckthorn, Rhamnus sp.) and a secondary summer host (soybean, Glycine max). Selection of soybean cultivars with different maturity groups may provide growers with a management tool for A. glycines, either directly through its effect on summer populations that cause economic damage or indirectly through its effect on the production of migrants that disperse to the primary host in fall. This study investigated the abundance and seasonality of A. glycines on soybean cultivars with different maturity rates in central Indiana. The abscission of soybean foliage occurred earlier for early maturing than late maturing cultivars, but no other consistent difference in development or yield was detected among the cultivars tested in this study. The abundance of aphids did not vary consistently among cultivars when soybean was most susceptible to economic damage. A laboratory assay evaluating the larviposition preference of A. glycines alate females, combined with a 7-yr survey documenting the colonization of buckthorn by winged aphids, indicated that the production of gynoparae on soybean began in mid-September and continued until leaf abscission. The abundance of aphids during this period was higher on late maturing cultivars than on early maturing cultivars in both 2006 and 2008, whereas no significant effect was detected in 2007. Altogether, these results suggest that planting early maturing soybean cultivars has little effect on damage by aphids on the current season crop but may reduce the number of fall migrants to the primary host.


Monoclonal antibody-based discrepancies between two-site immunometric tests for lutropin.
We compared five two-site immunometric assays (including three commercially available kits) for measuring lutropin (LH) in serum. Four of the assays involved monoclonal antibodies directed against the alpha-beta dimer, intact LH; these assays measured significantly lower concentrations of LH in 19 (out of 83) samples than did a commercial method not involving such antibodies. In five serum samples, two of the intact LH assays failed to detect any significant immunoreactivity above the detection limit. Findings of normal in vitro LH bioactivity in these samples did not confirm the low immunoreactivity of the intact LH assays. The inability of these assays to detect bioactive LH creates confusion in daily, routine testing as well as in research monitoring of bio/immuno ratios. The data presented here confirm our previous findings (Clin Chem 1991;37:333-40) and emphasize the need to avoid the use of monoclonal antibodies specific for the intact LH dimer.


In vivo evaluation of NM441, a new thiazeto-quinoline derivative.
NM441 is a lipophilic prodrug of a new thiazeto-quinoline carboxylic acid derivative NM394, and when it is administered orally it is readily absorbed and hydrolyzed to its parent compound. After oral administration of NM441 at a dose of 20 mg/kg to dogs, the peak concentration of NM394 in plasma was 2.39 micrograms/ml, whereas it was 0.63 micrograms/ml for NM394 administered alone. The in vivo activity of NM441 was compared with those of ciprofloxacin, ofloxacin, and enoxacin in mouse protection studies. NM441 was as effective as ofloxacin and was twice as effective as ciprofloxacin against systemic infection with Staphylococcus aureus. Against infections with streptococci, NM441 was two to three times as effective as ofloxacin and five times as effective as ciprofloxacin. Against infection with Escherichia coli, NM441 was as effective as ciprofloxacin and ofloxacin, but against infections with Klebsiella pneumoniae, Serratia marcescens, and Pseudomonas aeruginosa, NM441 was two to four times as effective as ciprofloxacin and ofloxacin. NM441 was three to seven times as effective as enoxacin in systemic infections. Against urinary tract infections with E. coli, NM441 reduced the number of bacterial CFU per gram of kidney by 1 to 2 log10 more and, with P. aeruginosa, by 1 to 6 log10 more than did ciprofloxacin and ofloxacin. Against respiratory tract infections with K. pneumoniae, NM441 was as effective as ofloxacin and was twice as effective as ciprofloxacin.