Single-Cell Transcriptomics Reveal Immune Mechanisms of the Onset and Progression of IgA Nephropathy.
IgA nephropathy (IgAN) is the leading cause of kidney failure due to an incomplete understanding of its pathogenesis. We perform single-cell RNA sequencing (RNA-seq) on kidneys and CD14+ peripheral blood mononuclear cells (PBMCs) collected from IgAN and normal samples. In IgAN, upregulation of JCHAIN in mesangial cells provides insight into the trigger mechanism for the dimerization and deposition of IgA1 in situ. The pathological mesangium also demonstrates a prominent inflammatory signature and increased cell-cell communication with other renal parenchymal cells and immune cells, suggesting disease progress from the mesangium to the entire kidney. Specific gene expression of kidney-resident macrophages and CD8+ T cells further indicates abnormal regulation associated with proliferation and inflammation. A transitional cell type among intercalated cells with fibrosis signatures is identified, suggesting an adverse outcome of interstitial fibrosis. Altogether, we systematically analyze the molecular events in the onset and progression of IgAN, providing a promising landscape for disease treatment.


Increased uptake of 5-FU in experimental liver tumours by simultaneous infusion of norepinephrine.
The effect of the simultaneous administration of norepinephrine and 5-fluorouracil (5-FU) on the uptake of radiolabelled 5-FU by liver tumours was studied in rats. Three different concentrations of 5-FU were used (15, 1.5 and 0.15 microgram/g body weight). The drugs were infused over a 30 min period via the hepatic artery, following cannulation of the gastroduodenal artery. The radioactivity in liver tumour, normal liver, lungs and intestines was estimated by liquid scintillation counting. At all concentrations tested, an increased uptake of radioactive 5-FU was found in the tumour when norepinephrine was infused. Tumour/liver ratios also increased significantly in all these cases. No significant differences were noted between norepinephrine infused and control animals in the radioactivity in normal liver, lungs and intestines. The effects noted were possibly due to changes in blood flow within the liver, but the possibility of a direct effect of norepinephrine on DNA metabolism is discussed.


Clinicopathological and Immunohistochemistry Study of a Long Survivor of Giant Cell Glioblastoma in a Patient With Neurofibromatosis 1: Case Report.
Glioblastoma multiforme (IDH wild type) is an aggressive glial tumor of astrocytic origin (WHO-grade 4) with a two-year median survival period. Patients who live more than three years are considered as long survivors. In this study, we present a long survivor of a known case of neurofibromatosis type 1 who developed GBM of the giant cell type at age 14 years, and now the patient, at age 28, has been cancer-free for more than 14 years.


Surface positive-charged modification of inorganic fillers to optimize lithium ion conductive pathways in composite polymer electrolytes for lithium-metal batteries.
The incorporation of inorganic fillers into composite polymer electrolytes (CPEs) is a common strategy to improve ionic conductivity. However, the high surface energy of inorganic fillers typically aggravates poor interfacial contact with polymer chains. Herein, we develop a surface positive-charge modification strategy for enhancing the intermolecular interaction of poly(ethylene oxide) (PEO) electrolytes with inorganic fillers and optimizing lithium ion (Li+) conductive pathways in CPEs. The SiO2 nanoparticles are coated with a polydopamine adhesive layer and then functionalized with a branched polyethyleneimine positively charged functional layer. Such surface modification not only effectively induces more amorphous structure into the PEO matrix but also promotes the dissociation of lithium salts and activates more free Li+ in the PEO to accelerate Li+ transport. The CPEs achieved a superior ionic conductivity of 6.12 × 10-5 S cm-1 at 30 °C. In addition, the modified fillers could induce the formation of a lithium fluoride (LiF)-rich solid-state interphase and correspondingly achieve excellent compatibility with Li metal. The Li symmetric battery using the as-prepared CPEs delivered stable Li plating/stripping performances over 3960 h under 0.2 mA cm-2. The resulting LiFePO4|Li battery has an excellent capacity retention of 92.8 % after 260 cycles at 0.5C and 60 °C.


Impact of substance use on the onset and course of early psychosis.
The strong comorbidity between psychosis and substance use is already identifiable in early psychosis, raising the question of the direction of the association between substance use and psychosis onset. It has long been considered that this association was explained by the self-medication hypothesis. This hypothesis has been recently challenged by several prospective studies carried out in population-based samples, showing a dose-response relationship between cannabis exposure and risk of psychosis. This association was independent from potential confounding factors such as exposure to other drugs and pre-existence of psychotic symptoms. As a large percentage of subjects from the general population is now exposed to this drug, even a small increase in the risk of adverse effects may have significant deleterious consequences for the health of the population. Hence, reducing exposure to cannabis may contribute to prevention of some incident cases of psychosis. Regarding prognosis, persistent substance misuse after the onset of psychosis has a deleterious impact on clinical outcome. Therapeutic programs for subjects with dual diagnosis should be implemented early in the course of psychosis to maximise their impact on the course of illness.


Analysis of phylogenetic and functional diverge in plant nine-cis epoxycarotenoid dioxygenase gene family.
During different environmental stress conditions, plant growth is regulated by the hormone abscisic acid (an apocarotenoid). In the biosynthesis of abscisic acid, the oxidative cleavage of cis-epoxycarotenoid catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED) is the crucial step. The NCED genes were isolated in numerous plant species and those genes were phylogenetically investigated to understand the evolution of NCED genes in various plant lineages comprising lycophyte, gymnosperm, dicot and monocot. A total of 93 genes were obtained from 48 plant species to statistically estimate their sequence conservation and functional divergence. Selaginella moellendorffii appeared to be evolutionarily distinct from those of the angiosperms, insisting the substantial influence of natural selection pressure on NCED genes. Further, using exon-intron structure analysis, the gene structures of NCED were found to be conserved across some species. In addition, the substitution rate ratio of non-synonymous (Ka) versus synonymous (Ks) mutations using the Bayesian inference approach, depicted the critical amino acid residues for functional divergence. A significant functional divergence was found between some subgroups through the co-efficient of type-I functional divergence. Our results suggest that the evolution of NCED genes occurred by duplication, diversification and exon intron loss events. The site-specific profile and functional diverge analysis revealed NCED genes might facilitate the tissue-specific functional divergence in NCED sub-families, that could combat different environmental stress conditions aiding plant survival.


Elucidating paramylon and other carbohydrate metabolism in Euglena gracilis: Kinetic characterization, structure and cellular localization of UDP-glucose pyrophosphorylase.
Many oligo and polysaccharides (including paramylon) are critical in the Euglena gracilis life-cycle and they are synthesized by glycosyl transferases using UDP-glucose as a substrate. Herein, we report the molecular cloning of a gene putatively coding for a UDP-glucose pyrophosphorylase (EgrUDP-GlcPPase) in E. gracilis. After heterologous expression of the gene in Escherichia coli, the recombinant enzyme was characterized structural and functionally. Highly purified EgrUDP-GlcPPase exhibited a monomeric structure, able to catalyze synthesis of UDP-glucose with a Vmax of 3350 U.mg-1. Glucose-1P and UTP were the preferred substrates, although the enzyme also used (with lower catalytic efficiency) TTP, galactose-1P and mannose-1P. Oxidation by hydrogen peroxide inactivated the enzyme, an effect reversed by reduction with dithiothreitol or thioredoxin. The redox process would involve sulfenic acid formation, since no pair of the 7 cysteine residues is close enough in the 3D structure of the protein to form a disulfide bridge. Electrophoresis studies suggest that, after oxidation, the enzyme arranges in many enzymatically inactive structural conformations; which were also detected in vivo. Finally, confocal fluorescence microscopy provided evidence for a cytosolic (mainly in the flagellum) localization of the enzyme.


Romosozumab: A Review in Postmenopausal Osteoporosis.
Romosozumab (Evenity®), a humanized monoclonal antibody, promotes bone formation and inhibits bone resorption by inhibiting sclerostin, a protein involved in the regulation of bone formation. Subcutaneous romosozumab is approved in several countries, including those of the EU for treating severe osteoporosis as well as in the USA for osteoporosis in postmenopausal women at high risk of fracture. In pivotal phase III trials (FRAME and ARCH), 12 months' once-monthly romosozumab 210 mg significantly reduced vertebral and clinical fracture risk versus placebo and oral alendronate in postmenopausal women with osteoporosis. After patients transitioned from romosozumab to 12-24 months of subcutaneous denosumab or oral alendronate, fracture risks were significantly improved versus placebo-to-denosumab and alendronate-only treatment. In these trials and a phase IIIb trial, romosozumab significantly increased bone mineral density (BMD) relative to placebo, alendronate and subcutaneous teriparatide at 12 months, with these benefits maintained 12-24 months after patients transitioned from romosozumab to alendronate or denosumab in pivotal trials. Romosozumab had a generally manageable tolerability profile. While further clinical experience is needed to more definitively establish its efficacy and safety, including its CV safety, romosozumab extends the treatment options in postmenopausal women with osteoporosis who have a high risk of fracture and in those who have failed or are intolerant to other available osteoporosis therapy.


112 Gb/s transmission with a directly-modulated laser using FFT-based synthesis of orthogonal PAM and DMT signals.
We report the experimental measurement of 112 Gb/s transmission back-to-back and through 12 km of S-SMF with a single directly-modulated laser (DML) using the novel modulation format Orthogonal PAM-DMT. This work demonstrates a record DML-based 112 Gb/s receiver sensitivity of -7.1 dBm at a BER of 10(-3), outperforming conventional PAM and DMT by approximately 2.5 dB.


Clinical course of duodenal mucosa-associated lymphoid tissue lymphoma: Comparison with gastric mucosa-associated lymphoid tissue lymphoma.
There are few cases of duodenal mucosa-associated lymphoid tissue (MALT) lymphoma reported in the literature, and little is known about the clinical course of this disease. Here, we aimed to characterize the clinical features of duodenal MALT lymphoma by comparison with gastric MALT lymphoma. Thirteen patients diagnosed with duodenal MALT lymphoma at Asan Medical Center from March 1997 to February 2017 were included in this retrospective study, along with patients with gastric MALT lymphoma, matched by age and sex at a 1:10 ratio. Median age of patients with duodenal MALT lymphoma was 49 (range 20-72) years, and 53.8% (7/13) were male. Comparison of patient characteristics indicated that Helicobacter pylori infection (46.2% vs 90.8%, P < 0.001) and lymph node metastasis (23.1% vs 5.4%, P = 0.049) rates differed between patients with duodenal and gastric MALT lymphoma. Overall complete remission (61.5% vs 86.2%, P = 0.021) and complete remission after initial H. pylori eradication therapy (50% vs 87.7%, P = 0.037) were significantly lower in patients with duodenal than gastric MALT lymphoma. Complications including bleeding, stricture, and transformation to high-grade lymphoma occurred in a total of seven patients (4.9%), with a higher incidence in patients with duodenal than gastric MALT lymphoma (38.5% vs 1.5%, P < 0.001). Duodenal MALT lymphoma is very rare, and treatment outcomes appear to be inferior to those of gastric MALT lymphoma.


A calcium-responsive transcription factor, CaRF, that regulates neuronal activity-dependent expression of BDNF.
Transcription of the brain-derived neurotrophic factor (BDNF) gene is regulated in a calcium- and neuron-selective manner; however, the mechanisms that underlie this selectivity are not known. We have characterized a new calcium-response element, CaRE1, that is required for activity-dependent transcription of BDNF exon III and have cloned a transcription factor, CaRF, that activates transcription from BDNF promoter III in a CaRE1-dependent manner. The transcriptional activity of CaRF is regulated in a calcium- and neuron-selective manner, suggesting that CaRF may confer selectivity upon the activity-dependent induction of BDNF exon III expression.


High level of serum complement 3 is a risk factor for vascular stenosis progression in TA patients receiving tocilizumab: a prospective observational study.
The IL-6R antibody tocilizumab has been proven effective in treating Takayasu arteritis (TA). However, some patients show silent vascular stenosis progression (VSP) despite treatment with tocilizumab. The aim of the study was to explore the related risk factors of VSP in patients treated with tocilizumab. Patients receiving tocilizumab were enrolled from the prospective living ongoing East China Takayasu Arteritis cohort. Their medical information was uniformly recorded with a homogenized evaluation method. Magnetic resonant angiography or computed tomographic angiography was employed to monitor VSP during the follow-up period, and Cox regression analysis was performed to explore the related risk factors. Thirty-eight patients were enrolled, among whom 18 (47.4%) experienced VSP, and seven and three patients experienced new and worsened vascular ischemic symptoms and events (VISE) during follow-up, respectively. The median period for VSP occurrence was 6.9 months during follow-up. Patients with VSP showed higher levels of baseline complement 3 (C3) than those in the patients without VSP. Multivariate Cox regression analysis revealed baseline C3 level (hazard ratio [HR] = 7.05, 95% confidence interval: 1.50-33.07, p = 0.013) was independently associated with VSP, with a cut-off value of 1.22 g/L. 47.4% of TA patients treated with tocilizumab would suffer VSP. A high C3 level is a risk factor for VSP in TA patients receiving tocilizumab, which may facilitate the option of tocilizumab in the future.


O-antigen and core carbohydrate of Vibrio fischeri lipopolysaccharide: composition and analysis of their role in Euprymna scolopes light organ colonization.
Vibrio fischeri exists in a symbiotic relationship with the Hawaiian bobtail squid, Euprymna scolopes, where the squid provides a home for the bacteria, and the bacteria in turn provide camouflage that helps protect the squid from night-time predators. Like other gram-negative organisms, V. fischeri expresses lipopolysaccharide (LPS) on its cell surface. The structure of the O-antigen and the core components of the LPS and their possible role in colonization of the squid have not previously been determined. In these studies, an O-antigen ligase mutant, waaL, was utilized to determine the structures of these LPS components and their roles in colonization of the squid. WaaL ligates the O-antigen to the core of the LPS; thus, LPS from waaL mutants lacks O-antigen. Our results show that the V. fischeri waaL mutant has a motility defect, is significantly delayed in colonization, and is unable to compete with the wild-type strain in co-colonization assays. Comparative analyses of the LPS from the wild-type and waaL strains showed that the V. fischeri LPS has a single O-antigen repeat composed of yersiniose, 8-epi-legionaminic acid, and N-acetylfucosamine. In addition, the LPS from the waaL strain showed that the core structure consists of L-glycero-D-manno-heptose, D-glycero-D-manno-heptose, glucose, 3-deoxy-D-manno-octulosonic acid, N-acetylgalactosamine, 8-epi-legionaminic acid, phosphate, and phosphoethanolamine. These studies indicate that the unusual V. fischeri O-antigen sugars play a role in the early phases of bacterial colonization of the squid.


Chitinophaga salinisoli sp. nov., isolated from saline soil.
A novel aerobic bacterium, designated strain LAM9153T, was isolated from a saline soil sample collected from Lingxian County, Shandong Province, China. Cells of strain LAM9153T were observed to be Gram-stain negative, non-motile, non-spore-forming and rod-shaped. The new isolate grew optimally at 30-35 °C, pH 7.0 and 0.5% of NaCl concentration (w/v). According to the phylogenetic analysis based on the 16S rRNA gene sequence, strain LAM9153T shares high similarity with Chitinophaga terrae Gsoil 238T (96.9%) and Chitinophaga niabensis JS 13-10T (95.9%), forming a subcluster with C. terrae Gsoil 238T, Chitinophaga cymbidii R156-2T, C. niabensis JS 13-10T and Chitinophaga soli Gsoil 219T in the phylogenetic tree. The major cellular fatty acids (> 10%) were identified as iso-C15:0, iso-C17:0 3-OH and summed features 3 (C16:1 ω6c and/or C16:1 ω7c). The predominant respiratory quinone was identified as menaquinone MK-7. The polar lipids consisted of phosphatidylethanolamine, aminophospholipid, three unidentified aminolipids and five unidentified lipids. The genomic DNA G+C content was determined to be 53.2 ± 1.6 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic data, strain LAM9153T is concluded to represent a novel species of the genus Chitinophaga, for which the name Chitinophaga salinisoli sp. nov. is proposed. The type strain is LAM9153T (= ACCC 19960T = JCM 30847T).


Real-time imaging of evoked activity in local circuits of the salamander olfactory bulb.
The encoding of olfactory information in the central nervous system (CNS) depends on spatially distributed patterns of activity generated simultaneously in many neuronal circuits. Optical neurophysiological recording permits analysis of neural activity non-invasively and with high spatial and temporal resolution. Here, a video method for imaging voltage-sensitive dye fluorescence in vivo is used to map neuronal activity in local circuits of the salamander olfactory bulb. The method permits the imaging of simultaneous ensemble transmembrane activity in real time. After electrical stimulation of the olfactory nerve, activity spreads centripetally from the sites of synaptic input to generate nonhomogeneous response patterns that are presumably mediated by local circuits within the bulbar layers. The results also show the overlapping temporal sequences of activation of cell groups in each layer. The method thus provides high resolution, sequential video images of the spatial and temporal progression of transmembrane events in neuronal circuits after afferent stimulation and offers the opportunity for studying ensemble events in other brain regions.


Association between travel times and food procurement practices among female supplemental nutrition assistance program participants in eastern North Carolina.
To examine associations between self-reported vehicular travel behaviors, perceived stress, food procurement practices, and body mass index among female Supplemental Nutrition Assistance Program (SNAP) participants. The authors used correlation and regression analyses to examine cross-sectional associations between travel time and distance, perceived stress, food procurement practices (grocery shopping frequency, fast-food consumption, home meal consumption), and body mass index among female SNAP participants aged 20-65 years (n = 215) in eastern North Carolina. There were positive associations (P < .05) between travel patterns and stress (r = 0.18 for average miles traveled), grocery shopping frequency (r = 0.16 for average miles traveled), and fast-food consumption (r = 0.19 for average miles traveled, r = 0.29 for average minutes traveled). SNAP education materials should focus on healthful interactions with the food environment, even when participants must travel long distances.


Interferon-gamma in the treatment of systemic sclerosis: a randomized controlled multicentre trial.
We report the results of a randomized controlled multicentre study on interferon-gamma (IFN-gamma) treatment of systemic sclerosis as determined by skin sclerosis, renal and other organ involvement, global assessment, subjective symptoms and quality of life. Forty-four patients were enrolled into the trial, 27 in the treatment group and 17 in the control group. All patients presented with type I or type II scleroderma. Twenty-nine patients (64%) finished the study. The mean duration of Raynaud's phenomenon and skin sclerosis was 15.3 and 10.8 years, respectively. The skin scores tended to improve in the treatment group (P > 0.05). Mouth aperture increased significantly from 38.5 to 47.7 mm in the treatment group (P < 0.001). Subanalysis of IFN-gamma treated patients with normalized skin sclerosis scores >/=1 showed significant improvement in both skin involvement and subjective symptoms (P < 0.05). Organ involvement improved in eight of 18 treatment patients and in three of 11 control patients. It worsened in three of 18 treatment patients and in four of 11 control patients. One control patient died due to cardiorespiratory failure during the study. No deterioration of renal function occurred during IFN-gamma treatment. There was a significant improvement in quality of life parameters in the control group but not in the treatment group. Plasma levels of neopterin increased significantly during IFN-gamma treatment but not in the control group, whereas N-terminal procollagen III peptide levels did not change in either group. There was a high frequency of mild to moderate influenza-like adverse events during IFN-gamma treatment. Only four of nine drop-out patients, however, experienced symptoms most probably associated with IFN-gamma treatment. We conclude that IFN-gamma therapy has mild beneficial effects on skin sclerosis and disease-associated symptoms in type I and II scleroderma. IFN-gamma treatment was associated with acceptable tolerability and did not induce major renal dysfunction in our patients.


High pressure liquid chromatographic analysis of in vivo metabolites of N-(substituted phenyl)-N'-(1,3,5-trimethyl pyrazole-4-yl)thioureas in rats.
The aim of this study was to investigate the in vivo metabolism of N-(substituted phenyl)-N'-(1,3,5-trimethylpyrazole-4-yl)thioureas (substrate) as model compounds in rats via HPLC. The substrates, N-(4-fluoro/chlorophenyl)-N'-(1,3,5-trimethylpyrazole-4-yl)thioureas (T2 and T3), and their possible metabolites were synthesized and the structures of the compounds were elucidated both by spectral and elemental analysis. Substrates were dissolved in 5% gum arabic and administered 100 mg/kg intraperitoneally (i.p.) in a volume of 0.1 ml/100 g. Blood samples were withdrawn before and at 30 min, 1, 2, 4, 8, 12 and 24 h post-dose. Chromatographic separation of the substrate and its metabolites was performed using a Hichrom chromasil C18 column (150 mm x 4.6 mm i.d., 5 microm particle size). The optimal composition of the mobile phase was achieved by using different mixtures of pure methanol and water. From the biotransformation of these thiourea compounds, N-dealkylation metabolites N-(4-fluoro/chloro-phenyl)-N'-(3,5-dimethylpyrazole-4-yl)thioureas (TI and T4) were identified together with unchanged substrate (T2 and T3) in the plasma by comparing them to reference standards via HPLC UV/DAD.


Direct activation of TGF-beta1 transcription by androgen and androgen receptor complex in Huh7 human hepatoma cells and its tumor in nude mice.
Importance of androgen for promotion of hepatocelullar carcinoma (HCC) has long been supported by clinical and experimental evidences. However, mechanisms involved in the carcinogenesis have not yet been fully elucidated. Moreover, unbalanced expression of TGF-beta1 during tumor progression results in prooncogenic rather than growth inhibition. To investigate the effect of androgen on transcriptional regulation of TGF-beta1, we isolated rat TGF-beta1 promoter, based on our previous report (GenBank AF249327), and examined regulation of its promoter activity by dihydrotestosterone in Huh7, LNCaP, and PC3 cells. Several putative transcription factor-binding sites were found, but no TATA box. When the full-length (-4784 to +68) and variously deleted promoter DNAs were evaluated, the promoter region spanning from -2732 to -1203 showed the highest activity towards dihydrotestosterone in a dose-dependent manner in both Huh7 and PC3 cells with androgen receptor (AR) expression. Putative androgen response sequence half site (5'-TGTCCT-3') was identified to be located within -1932 to -1927, proved by mutant (5'-AGACCT-3') analysis and chromatin immunoprecipitation (ChIP) assay. AR mediated upregulation of TGF-beta1 expression was confirmed by HCC developed in nude mice with AR-overexpressed Huh7-cells. This work presents in vivo and in vitro evidences of activation of TGF-beta1 expression by androgen and AR, and implicates the modulation of hepatocarcinogenesis by AR through the regulation of TGF-beta1 expression.


The temporal dimensions of anticonvulsant action of some newer benzodiazepines against metrazol induced seizures in mice and rats.
In a time-distribution study, the anticonvulsant effects of four benzodiazepine compounds were compared with those of three standard antiepileptics against metrazol-induced seizures in mice and rats. Ethosuximide and trimethadione had the shortest duration of action in mice, but protected the rats up to 6 hr. Phenobarbitone, diazepam, flurazepam and nitrazepam protected the mice up to 12 hr, but the rats were effectively protected only up to 3-4 hr. Clonazepam, the most potent and effective agent, protected the mice from clonic-tonic seizures up to 18-20 hr and the rats up to 6-7 hr. Comparison of the PD50 from clonic seizure at the peak-effect hours revealed that the benzodiazepines were 16 to 96 times more potent than phenobarbitone on a molar basis, while phenobarbitone itself was 12 to 26 times more potent than ethosuximide and trimethadione. Tonic seizures and mortality were largely suppressed by all drugs until 18-20 hr in mice and 6-7 hr in rats. Seizure latency and mortality patterns varied from drug to drug but not in a dose-dependent manner.


Crystallization and preliminary diffraction data for horse heart metmyoglobin.
Reddish-brown crystals of metmyoglobin from horse heart have been obtained by both the hanging drop and batch crystallization methods in the space group P2(1), having a = 64.3 A, b = 28.9 A and c = 35.9 A, with beta = 107.1 degree. Morphologically similar crystal forms have been obtained for three derivatives of horse heart myoglobin having modified heme prosthetic groups.


Embryopathic risks of cigarette smoking.
Cigarette smoking has been associated with infertility, a higher frequency of spontaneous abortions, intrauterine growth retardation and an increased incidence of birth defects. Nicotine is the major pharmacological component of cigarette smoke. Its potential reproductive and teratological risks are critically reviewed.


RAB5A is associated with genes involved in exosome secretion: Integration of bioinformatics analysis and experimental validation.
Exosomes, as cell-cell communicators with an endosomal origin, are involved in the progression of various diseases. RAB5A, a member of the small Rab GTPases family, which is well known as a key regulator of cellular endocytosis, is expected to be involved in exosome secretion. Here, we found the impact of RAB5A on exosome secretion from human hepatocellular carcinoma cell line using a rapid yet reliable bioinformatics approach followed by experimental analysis. Initially, RAB5A and exosome secretion-related genes were gathered from bioinformatics tools, namely, CTD, COREMINE, and GeneMANIA; and published papers. Protein-protein interaction (PPI) was then constructed by the Search Tool for Retrieval of Interacting Genes (STRING) database. Among them, several genes with different combined scores were validated by the real-time quantitative polymerase chain reaction (RT-qPCR) in stable RAB5A knockdown cells. Thereafter, to validate the bioinformatics results functionally, the impact of RAB5A knockdown on exosome secretion was evaluated. Bioinformatics analysis showed that RAB5A interacts with 37 genes involved in exosome secretion regulatory pathways. Validation by RT-qPCR confirmed the association of RAB5A with candidate interacted genes and interestingly showed that even medium to low combined scores of the STRING database could be experimentally valid. Moreover, the functional analysis demonstrated that the stable silencing of RAB5A could experimentally decrease exosome secretion. In conclusion, we suggest RAB5A as a regulator of exosome secretion based on our bioinformatics approach and experimental analysis. Also, we propose the usage of PPI-derived from the STRING database regardless of their combined scores in advanced bioinformatics analysis.


A UK consensus statement on thromboprophylaxis for autologous breast reconstruction.
Microsurgical breast reconstruction accounts for 22% of breast reconstructions in the UK. Despite thromboprophylaxis, venous thromboembolism (VTE) occurs in up to 4% of cases. Using a Delphi process, this study established a UK consensus on VTE prophylaxis strategy, for patients undergoing autologous breast reconstruction using free-tissue transfer. It captured geographically divergent views, producing a guide that reflected the peer opinion and current evidence base. Consensus was ascertained using a structured Delphi process. A specialist from each of the UK's 12 regions was invited to the expert panel. Commitment to three to four rounds of questions was sought at enrollment. Surveys were distributed electronically. An initial qualitative free-text survey was distributed to identify likely lines of consensus and dissensus. Each panelist was provided with full-text versions of key papers on the topic. Initial free-text responses were analyzed to develop a set of structured quantitative statements, which were refined via a second survey as a consensus was approached. The panel comprised 18 specialists: plastic surgeons and thrombosis experts from across the UK. Each specialist completed three rounds of surveys. Together, these plastic surgeons reported having performed more than 570 microsurgical breast reconstructions in the UK in 2019. A consensus was reached on 27 statements, detailing the assessment and delivery of VTE prophylaxis. To our knowledge, this is the first study to collate current practice, expert opinion from across the UK, and a literature review. The output was a practical guide for VTE prophylaxis for microsurgical breast reconstruction in any UK microsurgical breast reconstruction unit.


Single-molecule studies of flavivirus envelope dynamics: Experiment and computation.
Flaviviruses are simple enveloped viruses exhibiting complex structural and functional heterogeneities. Decades of research have provided crucial basic insights, antiviral medication and moderately successful gene therapy trials. The most infectious particle is, however, not always the most abundant one in a population, questioning the utility of classic ensemble-averaging virology approaches. Indeed, viral replication is often not particularly efficient, prone to errors or containing parallel routes. Here, we review different single-molecule sensitive fluorescence methods that are employed to investigate flaviviruses. In particular, we review how (i) time-resolved Förster resonance energy transfer (trFRET) was applied to probe dengue envelope conformations; (ii) FRET-fluorescence correlation spectroscopy to investigate dengue envelope intrinsic dynamics and (iii) single particle tracking to follow the path of dengue viruses in cells. We also discuss how such methods may be supported by molecular dynamics (MD) simulations over a range of spatio-temporal scales, to provide complementary data on the structure and dynamics of flaviviral systems. We describe recent improvements in multiscale MD approaches that allowed the simulation of dengue particle envelopes in near-atomic resolution. We hope this review is an incentive for setting up and applying similar single-molecule studies and combine them with MD simulations to investigate structural dynamics of entire flavivirus particles over the nanosecond-to-millisecond time-scale and follow viruses during infection in cells over milliseconds to minutes.


Multimodality evoked potentials and electrically elicited blink reflex in optic neuritis.
Pattern shift visual evoked potentials, brain stem auditory evoked potentials, spinal and scalp recorded somatosensory evoked potentials, and electrically elicited blink reflexes were investigated in 32 patients with isolated optic neuritis. Eleven patients were shown to have one additional lesion in the central nervous system outside the optic nerve. Therefore, cases with optic neuritis of unknown origin should be considered as possible cases of multiple sclerosis.


Validated capillary electrophoresis method for the determination of atropine and scopolamine derivatives in pharmaceutical formulations.
The simultaneous determination of atropine and scopolamine derivatives, which have similar structures, was investigated by using capillary zone electrophoresis. The effects of buffer pH, buffer concentration and hydroxypropyl-beta-cyclodextrin concentration on migration time and resolution of the investigated compounds were systematically studied. The selected electrophoretic buffer consisted of a 80 mM sodium citrate pH 2.5, containing 2.5 mM hydroxypropyl-beta-cyclodextrin as the complexing agent. Quantitative analysis was validated by testing the reproducibility of the method, giving a relative standard deviation less than 1 and 2% for the intermediate precision of migration times and peak area ratios, respectively. The linearity of the method was assessed between 50 and 150% of the theoretical content (coefficient of correlation greater than 0.99). The proposed method was found to be suitable and accurate for the determination of these basic drugs in pharmaceutical preparations.


Application of creatinine-based eGFR equations in Chinese septuagenarians and octogenarians.
The utilization of creatinine-based estimated glomerular filtration rate (eGFR) equations in the adult population is acknowledged. Nevertheless, the appropriateness of creatinine-based eGFR in septuagenarians and octogenarians is debatable. This study evaluates the creatinine-based equations in Chinese septuagenarians and octogenarians cohorts. This study employed a retrospective design, utilizing a review of the hospital medical records system to identify 347 hospitalized participants within the Division of Geriatrics or the Division of Nephrology. These participants underwent renal dynamic imaging with 99 m Tc-DTPA and serum creatinine testing. Comparison of the equations was performed, including the full age-spectrum equation (FAS-Cr equation), European Kidney Function Consortium equation (EKFC equation), Chronic Kidney Disease Epidemiology Collaboration equation for Asian (Asian CKD-EPI equation), Xiangya equation, and Lund-Malmö revised equation (LMR equation). Most equations tended to underestimate GFR. The FAS-Cr equation had the smallest interquartile range (IQR), while the Asian CKD-EPI equation (mGFR ≥ 30) and Xiangya equation (mGFR < 30) had the biggest IQRs. The FAS-Cr equation had the highest overall P30 of 63.98%, while the Asian CKD-EPI equation had the highest P30 of 75.64% in mGFR ≥ 60. The Xiangya equation, on the other hand, reported the lowest P30 of 36.36% in mGFR < 30. We discovered similar patterns in root-mean-square error (RMSE) as P30. GFR category misclassification rates in the entire cohort ranged from 46.11 to 49.86% for all equations. The FAS-Cr equation exhibited an advantage in octogenarians over other equations in the GFR category misclassification with mGFR lower than 60 ml/min/1.73 m2. None of the creatinine-based equations in this study could perform well regarding precision, accuracy, and CKD stages' classification for the Chinese elderly. Nevertheless, the FAS-Cr equation should be suitable for octogenarians with mGFR lower than 60 ml/min/1.73 m2.


Photochemical localization of the semotiadil binding region within the cardiac Ca2+ channel alpha1 subunit. Comparison with the skeletal muscle counterpart.
We have previously identified the binding region of a new Ca2+ antagonist semotiadil in the skeletal muscle Ca2+ channel. To the same semotiadil derivatives, the cardiac counterpart showed distinct and different binding characteristics: semotiadil and its photoaffinity analog D51-4700 inhibited [3H]PN200-110 binding to cardiac membrane preparations with IC50 values of 13-20 microM, which are 10 times higher than those in skeletal muscle. Hill slopes of the binding inhibition were 0.94-1.0 for the cardiac channels compared to 0.63-0.67 for the skeletal muscle channels. A possible explanation for the difference is that the semotiadil binding site is differently conferred in cardiac and skeletal muscle Ca2+ channels. To reveal this within the primary structure, photoaffinity labeling of cardiac membranes was employed. [3H]D51-4700 was photo-incorporated in several polypeptides but only the alpha1 subunit of the Ca2+ channel was photolabeled in a specific manner. Antibody mapping of the [3H]D51-4700-labeled alpha1 subunit with several anti-peptide antibodies revealed that the labeled site was located solely in a peptide fragment between Cys1461 and Lys1529. This region encompasses the labeled site of skeletal muscle, but contains several non-identical amino acid residues, which may participate in expressing different binding characteristics between the two muscle type Ca2+ channels.


Corrigendum: Serum CYR61 is associated with airway inflammation and is a potential biomarker for severity in chronic obstructive pulmonary disease.
[This corrects the article DOI: 10.3389/fmed.2021.781596.].


Preprophase bands in a suspension culture of the monocot Spartina pectinata.
Continuous suspension cultures of the marsh grass Spartina pectinata grow as either unorganized colonies or files of cells. Immunofluorescence of tubulin revealed microtubule (MT) structures similar to those encountered in meristematic cells, including cortical microtubule (MT) bands in some interphase cells and in all prophase cells. These MT bands were judged to be preprophase bands (PPBs) on the basis of their temporal appearance in the cell cycle and their position and orientation relative to division planes. Although PPBs are widely thought to be associated with organized tissues and polarized divisions, there are reports of PPBs in suspension cultures of four dicot species. This is the first report of a PPB in suspension cultures of a monocot species.


High-throughput genetic screening of meiotic commitment using fluorescence microscopy in Saccharomyces cerevisiae.
Simple genetic screens in budding yeast have identified many conserved meiotic regulators. However, the identification of genes involved in specific steps of meiosis may require a more complex genetic screen that allows visualization of meiosis. Here, we describe a high-throughput protocol using fluorescence microscopy to systematically screen an overexpression library to identify genes involved in meiotic commitment. We also explain how this protocol can be adapted for identifying proteins that function at different stages of meiosis. For complete details on the use and execution of this protocol, please refer to Gavade et al. (2022).


Comparison of SARS-CoV-2 detection from combined nasopharyngeal/oropharyngeal swab samples by a laboratory-developed real-time RT-PCR test and the Roche SARS-CoV-2 assay on a cobas 8800 instrument.
Although several assays have been developed to detect SARS-CoV-2 RNA in clinical specimens, their relative performance is unknown. The concordance between the cobas 8800 SARS-CoV-2 and a laboratory developed (LD) reverse transcriptase-polymerase chain reaction (RT-PCR) assay was assessed on 377 combined nasopharyngeal/oropharyngeal swabs in Hanks medium. The positive and negative agreement between these assays were 99.3 % (95 % CI, 97.3-99.9) and 77.1 % (95 % CI, 67.7-84.4), respectively, for an overall agreement of 93.6 % (95 % CI, 90.7-95.7) beyond random chance (kappa of 0.82, 95 % CI, 0.75-0.85). Of the 22 samples positive by cobas SARS-CoV-2 only, 9 were positive only for ORF-1 gene and had Cycle thresholds (Ct) > 35.1, 8 were positive only for the E gene with Ct > 35.5 and 5 were positive for both targets with Ct > 33.9. Samples positive only with the cobas assay were more often positive with only one gene target (77.3 %) than samples positive in both assays (16.9 %, p < 0.0001). Ct values in the cobas SARS-CoV-2 assay were significantly higher in the 279 samples testing positive in both assays (32.9 %, 95 % CI 32.3-33.6) compared to the 22 samples with discordant results (36.6 %, 95 % CI 36.2-37.1; p = 0.0009). An excellent correlation (r2 = 0.98) was obtained between Ct values of the ORF-1 and E targets in the cobas assays and a good correlation was obtained between LD RT-PCR test and cobas SARS CoV-2 ORF-1 target (r2 = 0.82). Our study demonstrated an excellent concordance between a LD RT-PCR and the cobas SARS-CoV-2 tests on the 8800 platform.


Alpha particle chemistry. On the formation of stable complexes between He2+ and other simple species: implications for atmospheric and interstellar chemistry.
The possibility that stable complexes may be formed between alpha particles (He2+) and small molecules is investigated using QCISD quantum mechanical calculations. Implications for their presence in the terrestrial atmosphere and/or in interstellar space are discussed.


Tumor Infiltration Depth as Predictor of Nodal Metastasis in Early Tongue Squamous Cell Carcinoma.
A retrospective longitudinal study was conducted to identify the cutoff value of infiltration depth for predicting the risk of lymph node metastasis of the neck in a well-defined population of surgically treated patients affected by stage T1 to T2 oral squamous cell carcinoma of the tongue. Sixty-seven patients were enrolled in this study. Forty-four patients (65.5%) had pN0 status and 23 (34.5%) had pN(+) status. Thirty-five positive lymph nodes were analyzed. The median follow-up for these patients was 51.4 months. The mean infiltration depth of the N-negative group was 2.4 mm; this was substantially different from the mean value observed in the N-positive group at 5.5 mm. A meaningful cutoff was identified at an infiltration depth value of 4 mm. Infiltration depth was identified as an important predictor for neck nodal status. In this specific population, mortality was associated with increasing tumor infiltration depth.


Differential increase in prevalence estimates of inadequate sleep among black and white Americans.
The National Health Interview Survey (NHIS) was used to ascertain whether increases in inadequate sleep differentially affected black and white Americans. We tested the hypothesis that prevalence estimates of inadequate sleep were consistently greater among blacks, and that temporal changes have affected these two strata differentially. NHIS is an ongoing cross-sectional study of non-institutionalized US adults (≥18 years) providing socio-demographic, health risk, and medical factors. Sleep duration was coded as very short sleep [VSS] (<5 h), short sleep [SS] (5-6 h), or long sleep [LS] (>8 h), referenced to 7-8 h sleepers. Analyses adjusted for NHIS' complex sampling design using SAS-callable SUDAAN. Among whites, the prevalence of VSS increased by 53% (1.5% to 2.3%) from 1977 to 2009 and the prevalence of SS increased by 32% (19.3% to 25.4 %); prevalence of LS decreased by 30% (11.2% to 7.8%). Among blacks, the prevalence of VSS increased by 21% (3.3% to 4.0%) and the prevalence of SS increased by 37% (24.6 % to 33.7%); prevalence of LS decreased by 42% (16.1% to 9.4%). Adjusted multinomial regression analysis showed that odds of reporting inadequate sleep for whites were: VSS (OR = 1.40, 95% CI = 1.13-1.74, p < 0.001), SS (OR = 1.34, 95 % CI = 1.25-1.44, p < 0.001), and LS (OR = 0.94, 95% CI = 0.85-1.05, NS). For blacks, estimates were: VSS (OR = 0.83, 95% CI = 0.60-1.40, NS), SS (OR = 1.21, 95% CI = 1.05-1.50, p < 0.001), and LS (OR = 0.84, 95% CI = 0.64-1.08, NS). Blacks and whites are characteristically different regarding the prevalence of inadequate sleep over the years. Temporal changes in estimates of inadequate sleep seem dependent upon individuals' race/ethnicity.


Sequential reduction and fixation for windswept pelvic ring injuries (LC3) corrects the deformity until healed.
The restoration, and fixation, of normal pelvic anatomy after a windswept type injury can be a difficult endeavor and our purpose is to describe a method to accomplish this. A stepwise and sequential technique was utilized to effectively reduce and stabilize this injury pattern. By first closing down the open disruption posteriorly and fixing with a partially threaded SI screw, a stable platform was created upon which to work from and subsequently distract and reduce the contralateral side via an anterior internal fixator (seven), external fixator (one), or plate (one). This was followed by a fully threaded SI screw in the compression side of the sacral fracture to hold the distraction. Nine consecutive patients with LC3 (61-B3.2) were included in the study with an average FU of 15 months. The Keshishyan deformity index revealed an initial mean deformity of 0.0456 which was corrected to 0.0170 (postop) and 0.0181 at latest follow up. This entailed an average correction of 62 % at the latest follow up. The follow-up group was significantly different from pre-op (p = 0.0040), but not post-op (p = 0.6833). Furthermore, post-op was significantly different from pre-op (p = 0.0089). This is an effective method of correcting and maintaining reduction until healing for this relatively rare and difficult-to-treat injury pattern.


Study of the prevalence of agents of sexually transmitted diseases in monkeys by the gene diagnosis method.
Carriership of agents of sexually transmitted diseases (Trichomonas, Chlamydia, Mycoplasma, Ureaplasma) is highly prevalent in healthy monkeys living in the Adler Breeding Center. The incidence of these microorganisms is appreciably higher in animals with gestoses and labor abnormalities in comparison with animals with normal genital function. Mixed infection caused by 2-4 agents is much more incident than monoinfection.


Case Report of a Left-sided Superior Vena Cava Causing Unique Positioning of Central Line.
Persistent left-sided superior vena cava is a rare congenital venous malformation. While often clinically asymptomatic, these variations in normal anatomy may give rise to complications with central venous catheter placement. We present a case of a 71-year-old male who presented to the emergency department with sepsis of unknown etiology. A right-sided central venous catheter was placed, and due to a persistent left-sided superior vena cava the post-procedure chest radiograph showed a uniquely positioned catheter tip within the left atrium. A persistent left-sided superior vena cava may lead to uniquely positioned catheter tip placement on post-procedural imaging. This case demonstrates the need to consider variants in normal venous anatomy, such as persistent left-sided superior vena cava, to aid with correct interpretation of post-procedure imaging findings.


Identification and characterization of an ectoine biosynthesis gene cluster from Aestuariispira ectoiniformans sp. nov., isolated from seawater.
An ectoine-producing bacterium, designated SWCN16T, was isolated from seawater and could be grown in a medium containing up to 12 % NaCl. A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SWCN16T belonged to the genus Aestuariispira, class Alphaproteobacteria, and shared the highest 16S rRNA gene sequence similarity of 96.8% with Aestuariispira insulae CECT 8488T. The phenotypic, chemotaxonomic, and genotypic characteristics findings of this study suggested that strain SWCN16T represented a novel species of the genus Aestuariispira. We propose the name Aestuariispira ectoiniformans sp. nov. for this species. Whole-genome sequencing analysis of the isolate revealed a putative ectABC gene cluster for ectoine biosynthesis. These genes were found to be functional using ectoine synthesis testing and S16-ectBAC cells, which were pET21a-ectBAC-transformed E. coli BL21 cells. We found that S16-ectBAC synthesized about 1.67 g/L extracellular ectoine and about 0.59 g/L intracellular ectoine via bioconversion at optimum conditions.


Comparative study of pituitary and bacteria-derived human growth hormone by monoclonal antibodies.
We have established hybridoma lines which secrete mouse monoclonal antibodies (Mabs) to human pituitary growth hormone, hGH. Using indirect competitive ELISA and indirect passive hemagglutination inhibition twelve different Mabs were characterized with regard to cross-reactivity with the hGH-related hormones, human chorionic somatomammotropin, hCS, and human prolactin, hPRL. The reactivity of these Mabs with pituitary hGH was compared to that with either bacterially-produced methionyl-hGH or to that of reduced and S-carboxymethylated hGH, which has an altered conformation. None of the Mabs reacted with hPRL. Four did not react with hCS whereas the others showed varying degree of cross-reactivity with hCS. All Mabs reacted more weakly with reduced and S-carboxymethylated hGH than with the native form of the hormone, which was not seen with conventional rabbit antisera to hGH. Thus in the case of hGH the Mabs are superior to conventional antisera in revealing small conformational differences. However the pituitary and bacterially-derived methionyl-hGH were indistinguishable as determined by the 12 Mabs.


The relationship between thermodynamics and the toxicity of gadolinium complexes.
The suitability of gadolinium complexes as magnetic resonance imaging contrast agents depends on a number of factors. A thermodynamic relationship to toxicity exists if one assumes that the chemotoxicity of the intact complex is minimal but that the toxicity of the components of the complex (free metal and uncomplexed ligands) is substantial. Release of Gd3+ from the complex is responsible for the toxicity associated with gadolinium complexes; this release appears to be a consequence of Zn2+, Cu2+, and Ca2+ transmetallation in vivo. This hypothesis is supported by acute toxicity experiments, which demonstrate that despite a 50-fold range of LD50 values for four Gd complexes, all become lethally toxic when they release precisely the same quantity of Gd3+, and by subchronic rodent toxicity experiments, which demonstrate a set of gross and microscopic findings similar to those known to be caused by Zn2+ deficiency. Finally, this hypothesis predicts that subtle changes in formulation can further enhance the intrinsic safety of these complexes.


The acute phase protein alpha-1-antitrypsin inhibits transferrin uptake in PLC/PRF/5 cells and increases release of hepatitis B virus surface antigen and alpha-fetoprotein.
The present study was designed to investigate whether the acute phase protein alpha-1-antitrypsin (alpha1-AT), which has an inhibitory effect on transferrin (tf) receptor-mediated iron uptake in K562 and THP1 cells, has a similar effect in PLC/PRF/5 cells. This hepatic cell line is of specific interest because it is infected with hepatitis B virus (HBV). Therefore, we addressed the additional question whether alpha1-AT has any effect on cellular protein synthesis and replication of HBV in PLC/PRF/5 cells. Cells were incubated with various concentrations of alpha1-AT, dexamethasone, IL-6 and desferrioxamine. HBs-AG, alpha-fetoprotein and albumin concentrations in culture media were measured using commercially available methods. For equilibrium inhibition binding experiments, cells were incubated with 85-182 pmol/l [125I]tf. To study the potential effect of alpha1-AT on DNA synthesis we measured the incorporation of [3H]thymidine into DNA. In equilibrium saturation binding experiments, [125I]tf bound to PLC/PRF/5 cells with K(D) 17.45 +/- 4.57 nM and a maximum density of binding sites of 267,285 +/- 39,915 sites/cell. In inhibition studies alpha1-AT demonstrated an apparently monophasic inhibition of [125I]tf to its receptor. At concentrations > 30 micromol/l alpha1-AT inhibited the growth of PLC/PRF/5 cells up to approximately 50%. The inhibitory effect of alpha1-AT on DNA synthesis was not as potent as that on growth. At the highest concentration of 100 micromol/l, alpha1-AT produced a 35% maximum inhibition of [3H]thymidine incorporation. Incubating PLC/PRF/5 cells with corticosteroids enhanced HBs-AG release significantly. Interestingly, alpha1-AT showed the same pattern of effects on cell metabolism and HBs-AG release as the corticosteroids. When we incubated the cells with 50 micromol/l alpha1-AT, alpha-fetoprotein production increased significantly and HBs-AG release almost doubled. We have to assume that there is a specific mechanism inducing HBs-AG release by alpha1-AT, as has been shown to be the case with steroids.


A Clinico-Radiological Study: Veria Technique of Cochlear Implant-A Study of 50 Cases.
Main limitation of classical technique is limited access to cochlea in terms of cochlear rotations and related structures, thus causing difficulty in electrode insertion. Veria technique allows full access to cochlea after raising tympanomeatal flap. To our best knowledge this is first clinic-radiological study for Veria technique studying distance between tympanic segment of facial nerve and posterior wall of external auditory canal (EAC) demonstrating facial nerve safety. Prospective study was done on 50 patients having bilateral sensori-neural hearing loss. Patients underwent cochlear implant surgery irrespective of age and gender. Preoperative high-resolution computed tomography temporal bone and magnetic resonance imaging head was done, distance between tympanic segment of facial nerve and posterior wall of EAC and basal turn angle were measured. Intraoperative NRT followed by orbito-meatal X-ray was done in post operative period. 25 (50%) were male, 25 (50%) female. 35 patients (70%) showed that the distance between tympanic segment of facial nerve and posterior wall of EAC was more than 3 mm with mean 4.41 mm (± 0.63 SD). Distance calculated was greater in older age group than younger group. Patient having inner ear malformation, mean was 3.96 mm (± .44 SD). Whereas patients having acquired disease, mean distance was 4.30 mm (± .47 SD). On gender comparison of basal turn angle score, no significant difference was observed male (54.34° ± 4.48°) versus female (55.66° ± 4.15°) (p = 0.282). Mean of basal turn angle (BTA) in ≤ 5 years age group was 54.89° ± 3.65°, in 6-10 years age group was 55.21° ± 5.23° and in age group ≥ 11 years was 54.93° ± 4° with no significant difference in mean value between the groups (p = 0.282). High jugular bulb was seen in 4 patients (2 in right side, 2 in left side), hypoplastic jugular bulb was seen in 10 patients (9 in left, 1 in right), otosclerosis in 2 patients. Veria technique is safe for facial nerve, as preoperatively distance between tympanic segment of facial nerve and posterior canal wall can be identified. It is suitable method for rotated cochlea (identified preoperatively through BTA) and deformed cochlea as it offers a wide visibility and accessibility as compared to posterior tympanotomy approach. BTA and distance between posterior canal wall of EAC and tympanic segment of facial nerve should be done in all cases to see any cochlear rotation and feasibility of surgery.


Targetable BRAF and RAF1 Alterations in Advanced Pediatric Cancers.
RAF family protein kinases signal through the MAPK pathway to orchestrate cellular proliferation, survival, and transformation. Identifying BRAF alterations in pediatric cancers is critically important as therapeutic agents targeting BRAF or MEK may be incorporated into the clinical management of these patients. In this study, we performed comprehensive genomic profiling on 3,633 pediatric cancer samples and identified a cohort of 221 (6.1%) cases with known or novel alterations in BRAF or RAF1 detected in extracranial solid tumors, brain tumors, or hematological malignancies. Eighty percent (176/221) of these tumors had a known-activating short variant (98, 55.7%), fusion (72, 40.9%), or insertion/deletion (6, 3.4%). Among BRAF altered cancers, the most common tumor types were brain tumors (74.4%), solid tumors (10.8%), hematological malignancies (9.1%), sarcomas (3.4%), and extracranial embryonal tumors (2.3%). RAF1 fusions containing intact RAF1 kinase domain (encoded by exons 10-17) were identified in seven tumors, including two novel fusions TMF1-RAF1 and SOX6-RAF1. Additionally, we highlight a subset of patients with brain tumor with positive clinical response to BRAF inhibitors, demonstrating the rationale for incorporating precision medicine into pediatric oncology. IMPLICATIONS FOR PRACTICE: Precision medicine has not yet gained a strong foothold in pediatric cancers. This study describes the landscape of BRAF and RAF1 genomic alterations across a diverse spectrum of pediatric cancers, primarily brain tumors, but also encompassing melanoma, sarcoma, several types of hematologic malignancy, and others. Given the availability of multiple U.S. Food and Drug Administration-approved BRAF inhibitors, identification of these alterations may assist with treatment decision making, as described here in three cases of pediatric cancer.


Understanding and modulating cyclin-dependent kinase inhibitor specificity: molecular modeling and biochemical evaluation of pyrazolopyrimidinones as CDK2/cyclin A and CDK4/cyclin D1 inhibitors.
Cyclin-dependent kinases (CDKs) play a key role in regulating the cell cycle. The cyclins, their activating agents, and endogenous CDK inhibitors are frequently mutated in human cancers, making CDKs interesting targets for cancer chemotherapy. Our aim is the discovery of selective CDK4/cyclin D1 inhibitors. An ATP-competitive pyrazolopyrimidinone CDK inhibitor was identified by HTS and docked into a CDK4 homology model. The resulting binding model was consistent with available SAR and was validated by a subsequent CDK2/inhibitor crystal structure. An iterative cycle of chemistry and modeling led to a 70-fold improvement in potency. Small substituent changes resulted in large CDK4/CDK2 selectivity changes. The modeling revealed that selectivity is largely due to hydrogen-bonded interactions with only two kinase residues. This demonstrates that small differences between enzymes can efficiently be exploited in the design of selective inhibitors.


Production and glycosylation of sperm constitutive proteins in the lizard Lacerta vivipara. Evolution during the reproductive period.
From epididymal fluid samples taken at three different times during the reproductive period (early April, late April, mid-May), the soluble proteins were separated with one dimensional electrophoresis on polyacrylamide gel. Their evolution was studied: firstly quantitatively, after staining with Coomassie blue, or, for one protein (the "L" protein), by immunodetection; secondly, according to their glycosylation after transfer to nitrocellulose and treatment with a set of labelled lectins: from Wheat germ, Ricinus communis, Lens culinaris, Asparagus pea or Canavalia ensiformis, with or without use of their specific inhibitor sugars. At least 15 proteins underwent a quantitative and/or qualitative evolution, mainly during the month of April. Protein "L" (19 kDa), which is androgen dependent and which fixates on to spermatozoa during their epididymal transit, appears to be little or not glycosylated. By contrast its accumulation in the epididymal canal increases considerably during the month of April. Five other proteins proved to be especially interesting because of their evolution during this same period, notably the MW 94, 67, 35, 29 and 25.5 kDa proteins. With the exception of the 67 kDa all the others increased quantitatively. All were decisively enriched in mannose or in methyl-mannoside residues. The proteins of MW 29 and 25.5 kDa were also enriched in galactose or N-acetyl galactosamine residues. These findings are of physiological significance since they are set up concomitantly with the acquisition of maximum motility of spermatozoa in the distal segment of the epididymis, and they coincide with a very great increase in testosteronemia.


Contraceptive choices for chronically ill adolescents.
Ten percent of teenagers have chronic medical illness and need effective contraception for pregnancy prevention. There are available safe and effective methods; however, the selection of a contraceptive may be challenging because of the complexity of the underlying medical illness. This article offers options for contraception for girls with various chronic medical conditions. Considerations of some of the newer contraception methods are discussed as future options for these girls.


Team-Based Learning in the Surgery Clerkship: Impact on Student Examination Scores, Evaluations, and Perceptions.
There is little evidence for effectiveness of team-based learning (TBL) in specialties such as Surgery. We developed and instituted TBLs in surgery clerkship and compared National Board of Medical Examiners (NBME) Surgery Subject Exam scores before and after implementation. We also analyzed students' feedback for their perception of TBLs. The TBLs were transitioned into the curriculum during the 2013-2014 academic year. The "before" and "after" implementation periods were 2011-2013 and 2014-2016, respectively. NBME Surgery Subject Examination scores at our institution and nationally were compared using the independent samples t test. Satisfaction with the clerkship was assessed with Association of American Medical Colleges Graduate Questionnaire data. Student feedback regarding TBL was gathered at the end of each surgery rotation and were analyzed for themes, both positive and negative. Mean NBME score was higher at our institution than nationally, both before (77.10 ± 8.75 vs. 75.20 ± 8.95, p = 0.032) and after (74.65 ± 8.0 vs. 73.10 ± 8.55, p = 0.071) TBL implementation. The mean score decreased following TBL implementation at our medical school (77.10 ± 8.75 vs. 74.65 ± 8.00, p = 0.039), but it was also lower nationally (75.20 ± 8.95 vs. 73.10 ± 8.55, p < 0.001). Further, students were more likely to rate the surgery clerkship as "good and/or excellent" on the Association of American Medical Colleges Graduate Questionnaire after TBL implementation (84.6% vs. 73.7%). In qualitative assessment, learners stated that TBLs were informative, helpful in studying for the shelf exam, and viewed them as an opportunity for interactive learning, and thus requested more TBLs. Areas for improvement included reading materials, directions, and organization of sessions. Student perception of TBL into our surgery clerkship has been both positive and provided feedback for improvement. In addition, our medical school graduates have continued to assess their surgery experience as "good" or "excellent" by a large majority. Concurrently, our NBME scores remain above the national mean. We believe our medical students benefit from a well-organized TBL and its active approach to learning during the surgery clerkship with no loss of fundamental surgery knowledge.


The role of hypoalbuminemia in the development of the TPN-associated cholestasis.
Forty-one patients undergoing total parenteral nutrition (TPN), were studied in a prospective manner. We found that patients with low serum albumin were more likely to develop cholestasis than patients with normal serum albumin. None of the patients with normal serum albumin developed cholestasis. Fifty-one per cent of patients with low serum albumin (< 3.0 g/dl) developed cholestasis (p < 0.05). In those who developed cholestasis there was a significant, correlation (r = 0.816, p < 0.001) between the serum albumin and the number of days after start of TPN when cholestasis appeared.


Segmental Gracilis Muscle Transplantation for Midfacial Animation in Möbius Syndrome: A 29-Year Experience.
Möbius syndrome is a complex congenital disorder of unclear cause involving multiple cranial nerves and typically presenting with bilateral facial and abducens nerves palsies. At The Hospital for Sick Children, Toronto, Ontario, Canada, microneurovascular transfer of free-muscle transplant is the procedure of choice for midfacial animation. The primary aim of this study was to investigate surgical outcomes of the procedure in terms of complications, secondary revisions, and smile excursion gains. A retrospective 29-year review was performed using patient records from a single tertiary care center. The authors included children with Möbius syndrome who had undergone facial animation surgery with a free segmental gracilis muscle transfer and microneurovascular repair between January 1, 1985, and August 31, 2014. Smile excursion measures were obtained using the Facial Assessment by Computer Evaluation-Gram on a subset of the included patients. A total of 107 patients undergoing 197 reconstructive procedures met inclusion criteria. Most reconstructions relied on the motor nerve branch to the masseter for innervation [n = 174 (88 percent)]. Thirteen complications were reported, of which six required surgical interventions. Three revision procedures were performed: scar revision, muscle repositioning, and removal of infected permanent suture material. The use of the motor nerve branch to the masseter resulted in good commissure excursion gains (average, 4.61 mm for bilateral cases and 9.34 mm for unilateral reconstructions). Midfacial animation with segmental gracilis muscle transfer for patients with Möbius syndrome provides gains in the amplitude and symmetry of oral commissure excursion and carries a reasonably low complication rate. Therapeutic, IV.


Comparison of Mineral Oil, Insecticidal, and Biopesticide Spraying Regimes for Reducing Spread of Three Potato virus Y Strains in Potato Crops.
In-field management of Potato virus Y (PVY) faces challenges caused by the changing availability and environmental acceptability of chemical agents to control aphid vectors of the virus and by proliferation of PVY strains with different symptoms and rates of spread. From 2018 to 2020, foliar spray treatments were compared in field experiments in New Brunswick, Canada, to measure effectiveness at reducing spread of PVYO, PVYN:O, and PVYNTN strains. Mineral oil, insecticide, combined oil and insecticide spray, and a biopesticide (i.e., LifeGard WG) were compared. Insecticide-only and mineral oil-only treatments were not effective, but several combined oil and insecticide treatments and biopesticide treatments significantly reduced PVY spread. The biopesticide was proportionately more effective with recombinant PVYN:O and PVYNTN strains, possibly by exciting the plant's hypersensitive resistance response, caused naturally only in cultivar 'Goldrush' by PVYO. Pesticide residue analysis showed that mineral oil increased the retention of pyrethroid insecticide in the potato foliage longer than with insecticide applied alone, which may explain the beneficial synergistic effect of combined sprays for reducing PVY spread. Tuber yields were generally unchanged in chemical insecticide treatments but were slightly lower in biopesticide treatment. The cost per PVY treatment was competitive across all effective treatments, including biopesticide; however, there was some revenue loss from lower yield with the biopesticide. This biopesticide is certified organic, however, and thus a small premium on the price for organic production could offset this yield deficit.


Predictors of moderate-to-vigorous physical activity (MVPA) in African American young adolescents.
African American adolescents residing in the South are at increased risk for obesity and physical inactivity, yet our understanding of potential influences is limited. Using an ecological framework, this study explored multilevel predictors (individual, family, home, and neighborhood environment) of moderate-to-vigorous physical activity (MVPA) among 116 African American adolescents (ages 12-16). Adolescents and their parents completed self-report surveys for hypothesized predictors. Youth physical activity was measured using accelerometry. In multiple regression models, decreased daily MVPA was associated with female sex (β = -24.27, p < 0.0001). Family social support (β = 1.07, p = 0.004) and adolescent self efficacy for PA (β = 6.89, p = 0.054) were positively associated with daily MVPA. Adolescent demographics along with family social support and self-efficacy influence younger African American adolescent physical activity. Further exploration of the complex interaction of multiple levels of influence is needed to develop appropriate interventions for this vulnerable group.


Microcylinders of endoplasmic reticulum in histiocytes in patients suffering from Degos' syndrome and dermatomyositis.
Rare intracytoplasmic membranous inclusions derived from the endoplasmic reticulum were observed in the histiocytes of one case of Dego' syndrome and one case of dermatomyositis. Cross-section and longitudinal sections made it possible to establish a three-dimensional model in which both microcylinders and layers of rolled-up double membranes appear.


Histaminergic nerve fibers in the median eminence and hypophysis of rats demonstrated immunocytochemically with antibodies against histidine decarboxylase and histamine.
Histaminergic fibers in the median eminence and hypophysis of rats were examined immunocytochemically with antibodies against histidine decarboxylase (HDC), the sole histamine-synthesizing enzyme, and against histamine itself. A similar distribution of immunoreactive fibers was observed with these two antibodies. In the median eminence, immunoreactive fibers were mainly located in the internal layer and could be traced to the posterior lobe of the hypophysis. A few fibers were detected in the external layer of the median eminence, but none in the anterior or intermediate lobe of the hypophysis. These observations suggest that neuronal histamine may take part in regulation of the hypothalamo-neurohypophysial neuroendocrine system in rats.


Kinetics and Mechanism of Selenium(IV) Oxidation by Aqueous Bromine Solution.
The bromine-selenite reaction at strongly acidic conditions was investigated by monitoring the absorbance-time traces at the isosbestic point of the bromine-tribromide system at a constant ionic strength (0.5 M adjusted by sodium perchlorate) and temperature. Despite the simplicity of the stoichiometry, the kinetics was found to be very complex. Although the formal kinetic orders of the reactants bromine and selenite are strictly 1, that of the hydrogen ion varies from -2 to less than -3 and notably depends on the initial bromide concentration as well. The bromide ion also inhibits the reaction, making the whole system as a sound example of efficient autoinhibition. We have clearly shown that the inhibitory effect of the bromide ion cannot be explained quantitatively by either exclusively considering the unreactivity of the tribromide ion over elemental bromine or driving the reaction via hypobromous acid formed from the well-known hydrolysis of bromine in aqueous solutions. Instead of that, bromonium ion transfer initiating equilibrium is suggested between the selenium(IV) and bromine species to produce bromide ion and SeO3Br- followed by the hydrolysis of this short-lived intermediate. This hydrolytic transformation was found to be catalytic with respect to hydroxide and bromide ions as well. We have also demonstrated that, among the wide variety of selenium species present in the acidic aqueous solution, the best result can be obtained by considering HSeO3 - as the kinetically active species toward bromine. The proposed mechanism containing 10 acid-base equilibria with known equilibrium constants, the above-mentioned initiating equilibrium, and the hydrolysis of SeO3Br- is able to fit all 49 kinetic absorbance-traces simultaneously, taking into account properly the most important characteristics of the measured data at strongly acidic conditions. Furthermore, this kinetic model was further extended by the direct reactions of hypobromous acid with selenium(IV) species suggested previously with reasonably modified rate coefficients to describe the pH dependence of the apparent second-order rate coefficients over the pH = 1-13 range, providing a useful tool to predict more accurately the kinetic behavior of selenium(IV) species in water treatment process conditions.


Low Rate of Invasive Fungal Infections During Induction and Consolidation Chemotherapy for Adults with De Novo Acute Myeloid Leukemia Without Anti-mold Prophylaxis: Single-Center 2002-2018 Empirical/Pre-emptive Approach.
Broad-spectrum antifungal prophylaxis is currently considered the standard of care for adults with de novo AML for the prevention of invasive fungal infections (IFIs), especially invasive pulmonary aspergillosis (IPA). Because fluconazole has been used in our center as anti-yeast prophylaxis, we sought to analyze in detail the incidence of IFIs over a 17-year period, as well as their impact on outcome. A standardized protocol of patient management, including serum galactomannan screening and thoracic CT-guided diagnostic-driven antifungal therapy, was used in all patients. A total of 214 consecutive adults with de novo AML who were treated in 3 CETLAM (Grupo Cooperativo para el Estudio y Tratamiento de las Leucemias Agudas y Mielodisplasias) protocols from 2002 to 2018 were included. The 90-day incidence of any IFI (including possible cases) was 11% (95% CI 4-15%), most cases occurred during induction chemotherapy (8%, 95% CI 4-12%), and most cases were probable/proven IPA (8%, 95% CI 3-13%). Developing an IFI during induction and consolidation had no impact on 1-year survival. A case-control study with 23 cases of IPA and 69 controls identified induction/re-induction chemotherapy, chronic pulmonary disease and age > 60 years/poor baseline performance status as potential pretreatment risk factors. The current study proves that inpatient induction and consolidation chemotherapy for de novo AML can be given in areas with "a priori" high-burden of airborne molds with fluconazole prophylaxis, while the selective use of anti-mold prophylaxis in patients at very high risk may further reduce the incidence of IFI in this specific clinical scenario.


Working within a spiritual framework.
Chris Barber, Visiting Lecturer (Learning Disabilities), Birmingham City University ( email@example.com ), offers tips for nurses on supporting patients in the spiritual aspects of their lives.


The feasibility of an inter-professional transitions of care service in an older adult population.
Older adults discharged from the Emergency Department (ED) are at high risk for medication interactions and side effects; examples of practice models addressing this transition of care are lacking. This was a prospective cohort study for adults in one of two urban community EDs. Patients ≥50 years of age discharged with at least one new, non-schedule II prescription medication were included. Patients had the option of three transitions of care services: 1) pharmacist-only with home delivery of discharge medications and full medication reconciliation, 2) pharmacist and home health care, including home delivery, medication reconciliation, and a visit from a home health nurse, or 3) either of the above without home delivery. Over seven months, 440 ED patients were screened. Of those, 43 patients were eligible, and three patients elected to join the study. All three patients selected pharmacy-only. Identified barriers to enrollment include the rate of schedule II prescriptions from the ED (53% of potential patients) and high patient loyalty to their community pharmacist. A pharmacy and home health care transitions of care program was not feasible at an urban community ED. While the pharmacist team identified and managed multiple medication issues, most patients did not qualify due to prescriptions ineligible for delivery. Patients did not want pharmacist or home health nurse involvement in their post ED visit care, many due to loyalty to their community pharmacy. Multiple barriers must be addressed to create a successful inter-professional transition of care model.


Upper Gastrointestinal Tract Motility Disorders in Women, Gastroparesis, and Gastroesophageal Reflux Disease.
This article reviews the sex differences in upper gastrointestinal (GI) motility for both healthy and common dysmotility conditions. It focuses on gastroesophageal reflux disease and other esophageal motor disorders for the esophagus and on gastroparesis and accelerated gastric emptying for the stomach. It also describes differences in upper GI motility signs and symptoms during each female hormonal stage (ie, menstrual cycle, pregnancy, perimenopause, menopause) for both healthy participants and those suffering from one of the aforementioned upper GI dysmotility conditions. More research still needs to be conducted to better understand sex differences in upper GI motility.


Mechanisms of allergic sensitization to foods: bypassing immune tolerance pathways.
The default response of the mucosal immune system to antigens derived from food is one of active immune tolerance carried out by regulatory T cells and induced by dendritic cells residing in the intestinal mucosa. This tolerance response must be inhibited or bypassed to generate allergic sensitization in experimental food allergy and this has been achieved by 3 main approaches: genetic modifications, experimental adjuvants, and bypassing oral tolerance by administering the antigen through alternative routes. This article discusses the implications of these approaches for understanding the mechanisms of sensitization to food allergens in human disease.


Expression of lipase-solubilized bovine liver microsomal cytochrome b5 in Escherichia coli as a glutathione S-transferase fusion protein (GST-cyt b5).
The gene coding for the lipase-solubilized bovine liver microsomal cytochrome b5 (cyt b5) was expressed in Escherichia coli BL21 cells as a glutathione S-transferase fusion protein (GST-cyt b5) using the constructed expression vector pGEX-cyt b). The GST-cyt b5 fusion protein can be matured in vivo as a holoprotein with heme incorporated into cyt b5 during the fermentation, and the purification procedures were simplified by using a one-step affinity column chromatography with glutathione-agarose gel. The fusion protein was characterized by its spectroscopic and electrochemical properties, the interaction between GST-cyt b5 and cyt c was also investigated. The results show that GST-cyt b5 fusion protein shares similar properties and functions to that of isolated cyt b5. Although cyt b5 and GST were fused together, the two partners have not made significant structural and functional alterations of their counterparts, the protein-protein interactions between them are apparently very weak. To our knowledge, the present study is the first report to express cyt b5 as a GST-cyt b5 fusion protein, which provides a good example for the in vivo maturation of a hemoprotein as a GST fusion protein and sheds new light on the protein-protein interactions within the GST fusion protein.


microRNA-199a-5p protects hepatocytes from bile acid-induced sustained endoplasmic reticulum stress.
Sustained endoplasmic reticulum (ER) stress has been linked to cell death and the pathogenesis of many liver diseases, including toxic liver, cholestasis, and infectious liver disease. The cellular pathways that attenuate hepatic ER stress have been the focus of many recent studies, but the role of microRNAs (miRNA) in this process remains unknown. Here, we report that one of the most abundant miRNAs in hepatocytes, miR-199a-5p, was elevated in both bile acid- and thapsigargin (TG)-stimulated cultured hepatocytes, as well as in the liver of bile duct-ligated mice. We identify the misfolded protein chaperone GRP78, as well as the unfolded protein response transducers endoplasmic reticulum to nucleus signaling 1 and activating transcription factor 6 as direct targets of miR-199a-5p, and show that endogenous miR-199a-5p represses the 3' untranslated regions (UTRs) of their mRNAs. Through gain-of-function and loss of function approaches, we demonstrate that the elevated miR-199-5p disrupts sustained ER stress and prevents hepatocytes from undergoing bile acid- or TG-induced cell death. Furthermore, we reveal that the transcription factor AP-1 is a strong positive regulator of miR-199a-5p. In brief, our study demonstrates that AP-1/miR-199a-5p and ER stress mediators form a feedback loop, which shields hepatocytes from sustained ER stress and protects the liver from injury. On the basis of these findings, we also suggest that the miRNA miR-199a-5p is a potential target for clinical approaches aiming to protect hepatocytes in liver disease.


Mechanistic insight into gel-induced aggregation of amorphous curcumin during dissolution process.
Amorphous curcumin (CUR) exhibited a decreased dissolution rate in comparison with the crystalline counterpart due to its gel formation during dissolution. The main purpose of the present study is to explore the mechanism of such gelation phenomenon. It was found that the dissolution of amorphous CUR and gel properties were influenced by the temperature and pH of the media. The formed gels were characterized by TPA, SEM, DSC, XRPD, FTIR and PLM. The results indicated that the gelation process led to the formation of a porous structure in which water molecules infiltrate, and entered into its supercooled liquid state with high viscosity when contacting aqueous media, accompanied by decreased Tg and crystalline transformation. In addition, mixing with hydrophilic excipients (such as hydrophilic silica) accelerated the gel formation of amorphous CUR, while the addition of hydrophobic excipients (such as hydrophobic silica and magnesium stearate) could effectively weaken and even eliminate the gelation, hence significantly improving its dissolution. Furthermore, according to contact angle measurement and fluorescence microscope observation, hydrophilic excipients were found to be able to accelerate water entering into the interior of amorphous CUR, hence facilitating the gelation, while hydrophobic excipients would hinder water infiltration into the powder and thus achieve degelation. In conclusion, it is important to recognize that the gelation potential of some amorphous materials should be considered in developing robust amorphous drug product of high quality and performance.


Ventrolateral medullary control of cardiovascular activity during muscle contraction.
An overview of the role of ventrolateral medulla (VLM) in regulation of cardiovascular activity is presented. A summary of VLM anatomy and its functional relation to other areas in the central nervous system is described. Over the past few years, various studies have investigated the VLM and its involvement in cardiovascular regulation during static muscle contraction, a type of static exercise as seen, for example, during knee extension or hand-grip exercise. Understanding the neural mechanisms that are responsible for regulation of cardiovascular activity during static muscle contraction is of particular interest since it helps understand circulatory adjustments in response to an increase in physical activity. This review surveys the role of several receptors and neurotransmitters in the VLM that are associated with changes in mean arterial pressure and heart rate during static muscle contraction in anesthetized animals. Possible mechanisms in the VLM that modulate cardiovascular changes during static muscle contraction are summarized and discussed. Localized administration of an excitatory amino-acid antagonist into the rostral portion of the VLM (RVLM) attenuates increases in blood pressure and heart rate during static muscle contraction, whereas its administration into the caudal part of the VLM (CVLM) augments these responses. Opioid or 5-HT1A receptor stimulation in the RVLM, but not in the CVLM, attenuates cardiovascular responses to muscle contraction. Furthermore, intravenous, intracerebroventricular or intracisternal injection of an alpha 2-adrenoceptor agonist or a cholinesterase inhibitor attenuates increases in blood pressure and heart rate during static muscle contraction. Finally, the possible involvement of endogenous neurotransmitters in the RVLM and the CVLM associated with cardiovascular responses during static muscle contraction is discussed. An overview of the role of the VLM in the overall cardiovascular control network in the brain is presented and critically reviewed.


Polymer nanocomposites with graphene-based hierarchical fillers as materials for multifunctional water treatment membranes.
Phase inversion of polymer casting mixtures filled with hierarchical functional nanostructures is proposed as a synthetic route for the design of multifunctional membranes. The study tested the hypothesis that by regulating the relative content of components representing different levels in the nanofiller hierarchy, the structure and additional functions of such membranes could be controlled separately. Exfoliated graphite nanoplatelets (xGnPs) decorated by Au nanoparticles (Au NPs), used as a model hierarchical nanofiller, were added to the casting mixture of polysulfone, N-Methyl-2-pyrrolidone and polyethylene glycol prior to forming the membrane by phase inversion. The resulting porous asymmetric nanocomposites were shown to be permselective and catalytically active ultrafiltration membranes that were more resistant to compaction, more permeable than xGnP-free membranes and at least as selective. By designing membrane compositions with different relative amounts of Au-decorated xGnPs and Au-free xGnPs, the structure (controlled by the loading of xGnPs) and catalytic activity (controlled by the loading of Au NPs) could be controlled largely independently.


Schwannoma of the skull base with intracranial extension.
A case is reported of solitary schwannoma involving the skull base with extension into the middle cranial fossa and parasellar region. The patient presented with an exophthalmos and ipsilateral facial numbness. The tumor was successfully excised via a two-stage procedure involving an initial intracranial and subsequent transmaxillary approach. The relevant literature on the presentation and treatment of schwannomas of the skull base with intracranial extension is also reviewed.


Thoracoscopic sympathectomy for axillary hyperhidrosis: the influence of T4.
Recent data suggest that severe compensatory sweating after sympathectomy for hyperhidrosis is more common than previously reported. In particular, T2-T4 sympathectomy for axillary hyperhidrosis leads to significantly more disabling sweating compared with T2-T3 sympathectomy for palmar hyperhidrosis. However, it is not known whether this is a result of the additional transection of the T4 segment or if patients with primary axillary hyperhidrosis are more prone to experience disabling compensatory sweating. A follow-up study by questionnaire was made of 100 consecutive patients who underwent thoracoscopic sympathectomy for axillary hyperhidrosis at two university hospitals. Patients underwent T2-T3 sympathectomy (n = 35) or T2-T4 sympathectomy (n = 65) depending on the surgeon's preference. The questionnaire was returned by 91% of patients after a median of 31 months. Compensatory sweating occurred in 90% of patients and was so severe in 61% that they often had to change clothes during the day. There were no significant differences in occurrence or severity of compensatory sweating between the two extents of sympathectomy. Surgical outcome, however, was significantly better after T2-T4 sympathectomy. In contrast with previous reports, the incidence of compensatory sweating was not significantly related to the extent of sympathectomy for axillary hyperhidrosis. This result suggests that patients with primary axillary hyperhidrosis are more prone to experience compensatory sweating. Although the majority of patients with axillary hyperhidrosis were satisfied after thoracoscopic sympathectomy, many regret the operation. Patients should undergo surgery only if medical treatments fail; and provided there is an indication, we recommend T2-T4 sympathectomy.


The effect of sulfhydryl and amino group reagents on human lymphocyte--sheep erythrocyte rosettes.
We have studied the influence of certain chemical groups, located on the surface of lymphocytes or sheep red blood cells, on the ability of these cells to form spontaneous rosettes. We found that fluorescamine, which reacts with amino groups, inhibits rosette formation, while p-chloromercuriphenylsulfonic acid, which binds to sulfydryl groups, increases the percentage of the rosette-forming lymphocytes. The possible mechanism of action of the above reagents is discussed.


Low calcium intake is associated with high plasma homocysteine levels in postmenopausal women.
Nutritional interventions targeting homocysteine remain controversial, and further nutritional research is warranted. We thus sought to explore the determinants of plasma homocysteine other than B-group vitamins. This cross-sectional study surveyed the nutritional status of 713 Japanese postmenopausal women using a semiquantitative food frequency questionnaire. Associations between total energy, protein, fat, carbohydrate, and vitamin A and K intakes and homocysteine were insignificant. Mean homocysteine in the second (536.1 ± 34.7 mg/day) and third (712.9 ± 115.6 mg/day) tertiles of calcium intake were lower than in the first tertile (379.6 ± 76.6 mg/day) by -0.57 nmol/mL (95 % confidence interval, -1.10 to-0.04, p = 0.04) and -1.18 nmol/mL (-1.76 to -0.60, p<0.01), respectively, after adjustment for lifestyle and clinical factors (trend p\0.01). Mean homocysteine in those with dietary calcium intake above the median (>536 mg/day) were lower regardless of the folic acid concentration; the differences were -1.59 nmol/mL (-2.33 to -0.85, p = 0.02) and -0.75 nmol/mL (-1.37 to-0.12, p<0.01) for the high (<7.8 ng/mL) and low folic acid groups, respectively. There was no significant association between calcium and folic acid (p = 0.08). In conclusion, further prospective research to confirm our findings is needed for the development of nutritional inventions targeting homocysteine.


Formocresol and ferric sulfate have similar success rates in primary molar pulpotomy. In carious primary molars does a pulpotomy performed with ferric sulphate, compared with formocresol, result in greater clinical/radiographic success?
Medline Ovid Library, the Cochrane Library, PubMed, Embase, Science Citation Index (SCI) and System for Information on Grey Literature in Europe (SIGLE). Reviews, full reports, or research abstracts of prospective, retrospective, comparative, and/or radiographic studies were included, while case reports and letters were excluded. A total of 13 studies (three randomized clinical trials and 10 clinical trials) contributed to the meta-analysis, one randomized clinical trial and one clinical trial were analyzed by the direct technique, and all 13 trials were analyzed by the indirect technique. Data from trials were divided into clinical and radiographic data, and separate statistical analyses were conducted using the direct technique. Odds ratios (ORs) were used to compare the relative success of ferric sulphate and formocresol. Data homogeneity was tested using the chi(2)-test of consistency on the ORs for each trial. Clinical data indicated that ferric sulphate was significantly more successful than formocresol (OR=1.95; CI=1.01-3.80). Radiographic data indicated no difference between medicaments (OR=0.90; CI=0.58-1.39). Medicaments did not differ with t-tests of clinical (P>0.10) and radiographic (P>0.50) data. In human carious primary molars with reversible coronal pulpitis, pulpotomies performed with either formocresol or ferric sulphate are likely to have similar clinical/radiographic success.


A feeding regime for the study of the interaction of ethanol and aging.
A suitable model for the study of the interaction of the aging process and prolonged treatment with ethanol is described. Animals were given 15% ethanol in the drinking water for up to 25 months. There was little disruption in the growth of the animals. Ethanol treatment resulted in a number of neurological and morphological changes some of which persisted throughout the treatment period. Both young and old animals displayed metabolic tolerance by the criteria of resistance to ethanol-induced hypothermia and increased metabolism of [14C]ethanol. The regime used has proved useful in the study of the interaction of ethanol and aging in the rat.


Dietary fat and fatty acid consumptions and the odds of asthenozoospermia: a case-control study in China.
Are dietary fat and fatty acid (FA) intakes related to the odds of asthenozoospermia? Plant-based fat consumption was associated with decreased asthenozoospermia odds, while the consumption of animal-based monounsaturated fatty acid (MUFA) was positively related to asthenozoospermia odds. Dietary fat and FA are significant ingredients of a daily diet, which have been demonstrated to be correlated to the reproductive health of men. However, to date, evidence on fat and FA associations with the odds of asthenozoospermia is unclear. The hospital-based case-control study was performed in an infertility clinic from June 2020 to December 2020. Briefly, 549 asthenozoospermia cases and 581 controls with normozoospermia were available for final analyses. We collected dietary data through a verified food frequency questionnaire of 110 food items. Asthenozoospermia cases were ascertained according to the World Health Organization guidelines. To investigate the correlations of dietary fat and FA consumptions with the odds of asthenozoospermia, we calculated the odds ratios (ORs) and corresponding 95% CIs through unconditional logistic regression models. Relative to the lowest tertile of consumption, the highest tertile of plant-based fat intake was inversely correlated to the odds of asthenozoospermia (OR = 0.68, 95% CI = 0.50-0.91), with a significant dose-response relation (OR = 0.85, 95% CI = 0.75-0.97, per standard deviation increment). Inversely, animal-based MUFA intake (OR = 1.49, 95% CI = 1.04-2.14) was significantly correlated to increased odds of asthenozoospermia, and an evident dose-response relation was also detected (OR = 1.24, 95% CI = 1.05-1.45, per standard deviation increment). Subgroup analyses showed similar patterns of associations to those of the primary results. Moreover, we observed significant interactions on both multiplicative and additive scales between animal-based MUFA and cigarette smoking. Selection bias and recall bias were unavoidable in any of the observational studies. As we failed to obtain the information of trans-fatty acid (TFA) consumption, the relation of TFA intake and asthenozoospermia odds was unclear. This study indicated that different sources of fat and FAs might exert different effects on the etiology of asthenozoospermia, and cigarette smoking could exacerbate the adverse effect of high animal-based MUFA intake on asthenozoospermia. Our findings provide novel evidence pertaining to the fields of prevention of asthenozoospermia through decreasing animal-derived fat and FA consumptions and smoking cessation. This work was supported by the JieBangGuaShuai Project of Liaoning Province, Natural Science Foundation of Liaoning Province, Clinical Research Cultivation Project of Shengjing Hospital, and Outstanding Scientific Fund of Shengjing Hospital. All authors have no conflict of interest to declare. N/A.


Changes in the facial profile following correction for mandibular prognathism.
A retrospective study of 33 patients was undertaken to investigate the changes occurring in the facial profile following surgical correction of mandibular prognathism. The parameters measured included surface area, shift of selected points, facial outline length and directional measurement in relation to the sella-nasion plane. The calculation of linear regression equations enabled prediction of the shift of selected soft tissue points and facial outline length. It is considered, that the analysis provides an accurate picture of the behaviour of the soft tissues in response to a given amount of bony movement following correction of the deformity.


Molecular characterisation of glutamate dehydrogenase gene defects in Japanese patients with congenital hyperinsulinism/hyperammonaemia.
Congenital hyperinsulinism and hyperammonaemia (CHH) is caused by dysregulation of glutamate dehydrogenase (GDH). We characterised the GDH gene in two Japanese patients with CHH. Patient 1 showed late-onset and mild hypoglycaemic episodes and mild hyperammonaemia, compared with patient 2. In GDH activity of lymphoblasts, patient 1 showed twofold higher basal GDH activity than control subjects and mild insensitivity for GTP inhibition. Patient 2 showed severe insensitivity for GTP inhibition, and similar allosteric stimulation by ADP in the controls. Genetic studies identified heterozygous and de novo L413V and G446D mutations in patients 1 and 2, respectively. COS cell expression study confirmed that both mutations were disease-causing gene. The insensitivity for GTP inhibition in L413V and G446D was emphasised in COS cell expression system as a result of the dosage effect of mutant GDH gene. L413V showed less impairment of GDH than G446D based on biochemical and genetic results, which was consistent with the clinical phenotype. Based on the structure of bovine GDH, G446D was located in GTP binding site of pivot helix and its surroundings, while L413V was located in alpha-helix of antenna-like structure. These different locations of mutations gave different effects on GDH enzyme. The antenna-like structure plays an important role in GDH activity.