Resilience to stress and risk of gastrointestinal infections.
Exposure to psychological stress can elicit a physiological response that may influence characteristics of the gastrointestinal mucosa, including increased intestinal permeability, in turn possibly increasing susceptibility to gastrointestinal infections. We investigated whether low stress resilience in adolescence is associated with an 'increased' risk of gastrointestinal infections in subsequent adulthood. Data were provided by Swedish registers for a cohort of 237 577 men who underwent military conscription assessment in late adolescence (1969-76). As part of the assessment procedure, certified psychologists evaluated stress resilience through semi-structured interviews. The cohort was followed from conscription assessment until 31 December 2009 (up to age 57 years). Cox regression assessed the association of stress resilience with gastrointestinal infections (n = 5532), with adjustment for family background measures in childhood and characteristics in adolescence. Peptic ulcer disease (PUD) in adulthood was modelled as a time-dependent covariate. Compared with high stress resilience, lower stress resilience was associated with a 'reduced' risk of gastrointestinal infections after adjustment for family background in childhood, characteristics in adolescence and PUD in adulthood, with hazard ratios (and 95% confidence intervals) of 0.88 (0.81-0.97) and 0.83 (0.77-0.88) for low and moderate stress resilience, respectively. Lower stress resilience in adolescence is associated with reduced risk of gastrointestinal infections in adulthood, rather than the hypothesized increased risk.


Actinobacterial 16S rRNA genes from freshwater habitats cluster in four distinct lineages.
We analysed the phylogenetic relatedness of 16S rRNA genes from freshwater bacteria affiliated with the class Actinobacteria. A polymerase chain reaction assay was developed to identify reliably rare Actinobacteria-related inserts within 16S rRNA gene clone libraries. In 18 libraries constructed from seven freshwater systems, altogether 63 actinobacterial sequence types were collected from a total of > 1800 clones. Sixty of the newly obtained sequences grouped within four distinct phylogenetic lineages. They constitute approximately 75% of the nearly complete sequences within these clusters that are presently available. A comparison with > 300 sequences from various soil habitats revealed that two of these monophyletic actinobacterial clades (acI and acII) almost exclusively harbour 16S rRNA sequence types from freshwaters and estuaries. This may indicate that such bacteria are not inoculated to freshwaters from terrestrial sources, but are autochthonous components of freshwater microbial assemblages. In contrast, sequence types from freshwaters, marine sediments and soils were clearly mixed in another of the actinobacterial lineages (acIV). Sequence divergence within acIV was the highest of all four lineages (88% minimum similarity), which potentially reflects its radiation across several habitat types. Within the freshwater lineages, groups of essentially identical sequence types were retrieved from geographically distant aquatic systems with strikingly different hydrological and limnological characteristics. This points to the necessity to investigate genotypic variability, in situ abundances and activities of these Actinobacteria in freshwater plankton in greater detail by cultivation-independent techniques.


Personal Cooling Garments: A Review.
Thermal comfort is of critical importance to people during hot weather or harsh working conditions to reduce heat stress. Therefore, personal cooling garments (PCGs) is a promising technology that provides a sustainable solution to provide direct thermal regulation on the human body, while at the same time, effectively reduces energy consumption on whole-building cooling. This paper summarizes the current status of PCGs, and depending on the requirement of electric power supply, we divide the PCGs into two categories with systematic instruction on the cooling materials, working principles, and state-of-the-art research progress. Additionally, the application fields of different cooling strategies are presented. Current problems hindering the improvement of PCGs, and further development recommendations are highlighted, in the hope of fostering and widening the prospect of PCGs.


Capillary electrophoresis-a high performance analytical separation technique.
Capillary electrophoresis (CE) is often one of the preferred techniques in pharmaceutical quality control and in clinical chemistry, particularly considering the high selectivity and lower costs compared to HPLC. The precision of CE is as good as in liquid chromatography (LC). The sample-throughput is high due to short analysis times. Efforts for sample pre-treatment are usually minor in CE. Urine and even blood plasma can be directly injected without further pre-treatment. After summarising the basic principles of CE, general strategies for method development are described to achieve selective, efficient, precise, fast, sensitive, and validated methods. Sample pre-treatment requirements are discussed. Standard buffer recipes, surfactants used in micellar electrokinetic chromatography (MEKC), chiral selectors, useful buffer additives, actions to deal with complex matrices, and aspects of validation have been collected. Other techniques that can be performed with CE instruments, such as capillary isoelectric focusing (CIEF), capillary isotachophoresis (CITP), and capillary electrochromatography (CEC), are briefly discussed.


[Bacterial lipopolsaccharides in pathogenesis of gynecological diseases and obstetric complications].
Analysis of literature data and author studies on the role of lipopolysaccharides (endotoxin) of Gram negative bacteria in women genital tract pathology and obstetric complications is presented. The role of endogenous infection associated with altered microecology of intestinal tract and vaginal biotopes of women in the development of endotoxinemia is discussed. The participation of endotoxin in embryo resorption, delay of intrauterine development and antenatal death of fetus, premature birth, pre-eclampsia, placental dysfunction is examined. The level of endotoxinema and pro-inflammatory cytokines is a marker of chronic endogenous infectious-inflammatory disease of various parts of genital tract with damage of a network of female reproduction system organs.


Site-specific regulation of gene expression by n-3 polyunsaturated fatty acids in rat white adipose tissues.
Dietary n-3 polyunsaturated fatty acids (n-3 PUFAs) limit abdominal fat depot hypertrophy. This could be due to regulation of the expression of proteins involved in adipose tissue metabolism. We investigated in vivo whether fatty acid synthase (FAS), hormone-sensitive lipase (HSL), lipoprotein lipase (LPL), phosphoenolpyruvate carboxykinase (PEPCK), CCAAT/enhancer binding protein alpha (C/EBP alpha), and leptin mRNA levels are affected in retroperitoneal (RP) and subcutaneous adipose tissues (SC) of rats fed n-3 PUFAs. For 4 weeks rats were fed high fat diets (20% fat) containing n-3 PUFAs given as eicosapentaenoic acid (EPA group), docosahexaenoic acid (DHA group), a mixture of these two fatty acids (MIX group), or native fish oil (FO group). A control group was fed with lard plus olive oil (LOO group). Final mean fat cell weight in RP ranged according to: LOO > or = EPA > or = DHA = FO = MIX. There was no difference in fat cell size of SC when comparing the LOO and MIX groups. The fatty acid compositions of RP and SC were similar and resemble that of dietary fat within each experimental group. In RP and compared to the LOO group, FAS, HSL, PEPCK, LPL, C/EBP alpha, and leptin mRNA levels decreased although not significantly in the EPA group, and were 40-75% lower in the DHA and MIX groups. mRNA levels were positively correlated to fat cell size in RP. In contrast, n-3 PUFAs had no effect on gene expression in SC. We conclude that n-3 PUFAs and mainly 22:6n-3 affect gene expression in a site-dependent manner in white adipose tissues via possible antiadipogenic effects.


Oropharyngeal microbiome profiled at admission is predictive of the need for respiratory support among COVID-19 patients.
The oropharyngeal microbiome, the collective genomes of the community of microorganisms that colonizes the upper respiratory tract, is thought to influence the clinical course of infection by respiratory viruses, including Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of Coronavirus Infectious Disease 2019 (COVID-19). In this study, we examined the oropharyngeal microbiome of suspected COVID-19 patients presenting to the Emergency Department and an inpatient COVID-19 unit with symptoms of acute COVID-19. Of 115 initially enrolled patients, 50 had positive molecular testing for COVID-19+ and had symptom duration of 14 days or less. These patients were analyzed further as progression of disease could most likely be attributed to acute COVID-19 and less likely a secondary process. Of these, 38 (76%) went on to require some form of supplemental oxygen support. To identify functional patterns associated with respiratory illness requiring respiratory support, we applied an interpretable random forest classification machine learning pipeline to shotgun metagenomic sequencing data and select clinical covariates. When combined with clinical factors, both species and metabolic pathways abundance-based models were found to be highly predictive of the need for respiratory support (F1-score 0.857 for microbes and 0.821 for functional pathways). To determine biologically meaningful and highly predictive signals in the microbiome, we applied the Stable and Interpretable RUle Set to the output of the models. This analysis revealed that low abundance of two commensal organisms, Prevotella salivae or Veillonella infantium (< 4.2 and 1.7% respectively), and a low abundance of a pathway associated with LPS biosynthesis (< 0.1%) were highly predictive of developing the need for acute respiratory support (82 and 91.4% respectively). These findings suggest that the composition of the oropharyngeal microbiome in COVID-19 patients may play a role in determining who will suffer from severe disease manifestations.


[Optimization of physico-chemical parameters of reaction media for determining the activity of RNA-polymerases of mollicutes].
RNA-polymerases of mollicutes differ considerably from certain species of bacteria in the temperature optimum of the activity manifestation. The activity of mollicute enzymes is considerably higher in the presence of manganese ions than in the presence of magnesium ions. They differ from Escherichia coli transcriptase in this character but are similar to the RNA-polymerase of lactic bacteria, hypothetic ancestors of acholeplasm. Sensitivity of RNA-polymerases to metals may be one of arguments explaining phylogeny of mycoplasms. It is established that for studying mollicute transcriptase the reacting mixture for examining the enzymic activity besides the major components should have the following parameters: pH 8.0; the MnCl concentration--8-10 mM; for form II of the A. laidlawii subsp. granulum 118 enzyme--4.5 mM; ammonium sulphate concentration--40 mM; for form II of st. 118-20 mM; the reaction should be conducted at the temperature of 27 degrees C for 30 min.


Multiple signaling pathways coordinate to induce a threshold response in a chordate embryo.
In animal development, secreted signaling molecules evoke all-or-none threshold responses of target gene transcription to specify cell fates. In the chordate Ciona intestinalis, the neural markers Otx and Nodal are induced at early embryonic stages by Fgf9/16/20 signaling. Here we show that three additional signaling molecules act negatively to generate a sharp expression boundary for neural genes. EphrinA signaling antagonizes FGF signaling by inhibiting ERK phosphorylation more strongly in epidermal cells than in neural cells, which accentuates differences in the strength of ERK activation. However, even weakly activated ERK activates Otx and Nodal transcription occasionally, probably because of the inherently stochastic nature of signal transduction processes and binding of transcription factors to target sequences. This occasional and undesirable activation of neural genes by weak residual ERK activity is directly repressed by Smad transcription factors activated by Admp and Gdf1/3-like signaling, further sharpening the differential responses of cells to FGF signaling. Thus, these signaling pathways coordinate to evoke a threshold response that delineates a sharp expression boundary.


Metallurgical consideration on the calibration curve for binary alloy samples in low-pressure argon laser-induced plasma spectrometry.
The composition dependence of the emission intensities was investigated in Cu-Ni as well as Ni-Zn binary alloy samples when a low-pressure argon laser-induced plasma was employed as the excitation source. The calibration curve in the Cu-Ni alloy system gave a linear relationship, implying that the selective evaporation of Cu or Ni atoms was caused less in those alloys having several chemical compositions. The Cu-Ni binary alloy has a solid solution phase all over the chemical compositions (all-proportional solid solution): Cu and Ni atoms form no intermetallic compounds but can occupy any positions of a face-centered cubic lattice without any particular interaction. This metallurgical structure would enable Cu and Ni atoms to be released from the sample surface individually, which means that the amount of ablation corresponds to the chemical composition of the alloy sample. For a comparison, a Ni-Zn binary alloy system was also investigated to find calibration curves yielding a nonlinear relationship, differing from those of the Cu-Ni alloy. The reason for this is that the Ni-Zn binary alloy has several metallurgical phases comprising different intermetallic compounds which would each vary the evaporation behavior of Zn. It should be paid attention in LIPS that the ablated composition after laser irradiation is sometimes different from the chemical composition, depending on the kinds of samples and their metallurgical structures.


Integrated Case-Based Applied Pathology (ICAP): a diagnostic-approach model for the learning and teaching of veterinary pathology.
Integrative Case-Based Applied Pathology (ICAP) cases form one component of learning and understanding the role of pathology in the veterinary diagnostic process at the Faculty of Veterinary Science, University of Sydney. It is a strategy that focuses on student-centered learning in a problem-solving context in the year 3 curriculum. Learning exercises use real case material and are primarily delivered online, providing flexibility for students with differing learning needs, who are supported by online, peer, and tutor support. The strategy relies heavily on the integration of pre-clinical and para-clinical information with the introduction of clinical material for the purposes of a logical three-level, problem-oriented approach to the diagnosis of disease. The focus is on logical diagnostic problem solving, primarily using gross pathology and histopathological material, with the inclusion of microbiological, parasitological, and clinical pathological data. The ICAP approach is linked to and congruent with the problem-oriented approach adopted in veterinary medicine and the case-based format used by one of the authors (PJC) for the teaching and learning of veterinary clinical pathology in year 4. Additionally, final-year students have the opportunity, during a diagnostic pathology rotation, to assist in the development and refinement of further ICAPs, which reinforces the importance of pathology in the veterinary diagnostic process. Evidence of the impact of the ICAP approach, based primarily on student surveys and staff peer feedback collected over five years, shows that discipline-specific learning, vertical and horizontal integration, alignment of learning outcomes and assessment, and both veterinary and generic graduate attributes were enhanced. Areas for improvement were identified in the approach, most specifically related to assistance in the development of generic teamwork skills.


Rural Melanoma Patients Have Less Surgery and Higher Melanoma-Specific Mortality.
Melanoma causes most skin cancer-related deaths, and disparities in mortality persist. Rural communities, compared to urban, face higher levels of poverty and more barriers to care, leading to higher stage at presentation and shorter survival in melanoma. To further evaluate these disparities, we sought to assess the association between rurality and melanoma cause-specific mortality and receipt of recommended surgery in a national cohort. Patients with primary non-ocular, cutaneous melanoma from the SEER database, 2000-2017, were included. Outcomes included melanoma-specific survival and receipt of recommended surgery. Rurality was based on Rural-Urban Continuum Codes. Variables included age, sex, race, ethnicity, income, and stage. Multivariate regression models assessed the effect of rurality on survival and receipt of recommended surgery. 103,606 patients diagnosed with non-ocular cutaneous primary melanoma met criteria during this period. 93.3% (n = 96620) were in urban areas and 6.7% (n = 6986) were in rural areas. On multivariate regression controlling for age, sex, race, ethnicity, and stage patients living in a rural area were less likely to receive recommended surgery (aOR .52, 95% CI: .29-.90, P = .02) and had increased hazard of melanoma-specific mortality (aHR 1.19, 95% CI: 1.02-1.40, P = .03) even after additionally controlling for surgery receipt. Using a large national cohort, our study found that rural patients were less likely to receive recommended surgery and had shorter melanoma cause-specific survival. Our findings highlight the importance of access to cancer care in rural areas and how this ultimately effects survival for these patients.


Augmented visual feedback increases finger tremor during postural pointing.
Physiological tremor in the upper limb of eight adults was examined during the performance of a unilateral pointing task under conditions where the visual feedback, limb used and target size were altered. All subjects were required to aim a hand-held laser pointer at a circular target 5.5 m away with the goal of keeping the laser emission within the centre of the target. Visual feedback was defined as either normal vision (NV) of their limb tremor, where the laser was switched off, or augmented vision (AV) where the laser was switched on. Postural tremor from the segments of the upper limb, forearm muscle EMG activity, and target accuracy measures were recorded and analysed in the time and frequency domains. Accuracy-tremor relations were assessed using cross correlation and linear regression. Results revealed a high degree of similarity in the general pattern of the tremor output seen for each limb segment across conditions with only scalar (amplitude) changes being seen as a function of the different constraints imposed. For any single condition the tremor amplitude increased from proximal to distal segments. The frequency profile for the tremor in any segment displayed two prominent frequency peaks (at 2-4 Hz and 8-12 Hz). A third, higher frequency peak (18-22 Hz) was observed in the index fingers only. Across all conditions significant coupling relations were observed only between the hand-finger and forearm-upper arm segment pairs. Altering the visual feedback was shown to have the greatest effect on limb tremor with increased tremor and EMG activity and decreased coupling being seen under AV conditions. In trying to reduce tremor output when the augmented feedback was provided novice subjects instead increased muscle activity which resulted in increased tremor. Overall these results indicate that the physiological tremor output observed in neurologically normal subjects is not simply the product of intrinsic oscillations but is influenced by the nature of the task being performed.


The transpeptidase PBP2 governs initial localization and activity of the major cell-wall synthesis machinery in E. coli.
Bacterial shape is physically determined by the peptidoglycan cell wall. The cell-wall-synthesis machinery responsible for rod shape in Escherichia coli is the processive 'Rod complex'. Previously, cytoplasmic MreB filaments were thought to govern formation and localization of Rod complexes based on local cell-envelope curvature. Using single-particle tracking of the transpeptidase and Rod-complex component PBP2, we found that PBP2 binds to a substrate different from MreB. Depletion and localization experiments of other putative Rod-complex components provide evidence that none of those provide the sole rate-limiting substrate for PBP2 binding. Consistently, we found only weak correlations between MreB and envelope curvature in the cylindrical part of cells. Residual correlations do not require curvature-based Rod-complex initiation but can be attributed to persistent rotational motion. We therefore speculate that the local cell-wall architecture provides the cue for Rod-complex initiation, either through direct binding by PBP2 or through an unknown intermediate.


Positive and negative selection on mammalian Y chromosomes.
Y chromosomes are genetically degenerate in most organisms studied. The loss of genes from Y chromosomes is thought to be due to the inefficiency of purifying selection in nonrecombining regions, which leads to the accumulation of deleterious mutations via the processes of hitchhiking, background selection, and Muller's ratchet. As the severity of these processes depends on the number of functional genes linked together on the nonrecombining Y, it is not clear whether these processes are still at work on the old, gene-poor mammalian Y chromosomes. If purifying selection is indeed less efficient in the Y-linked, compared to the X-linked genes, deleterious nonsynonymous substitutions are expected to accumulate faster on the Y chromosome. However, positive selection on Y-linked genes could also increase the rate of amino acid-changing substitutions. Thus, the previous reports of an elevated nonsynonymous substitution rate in Y-linked genes are still open to interpretation. Here, we report evidence for positive selection in two out of three studied mammalian Y-linked genes, suggesting that adaptive Darwinian evolution may be common on mammalian Y chromosomes. Taking positive selection into account, we demonstrate that purifying selection is less efficient in mammalian Y-linked genes compared to their X-linked homologues, suggesting that these genes continue to degenerate.


The prevalence of Helicobacter pylori infection and the status of gastric acid secretion in patients with Barrett's esophagus in Japan.
The acidity of the refluxate into the esophagus is a key factor for the pathogenesis of gastroesophageal reflux disease. Helicobacter pylori (H. pylori) infection can influence gastric acid secretion. We have reported that H. pylori infection prevents reflux esophagitis by decreasing gastric acid secretion in Japanese patients, but the role of this organism in Barrett's esophagus is unclear. The aim of this study was to investigate the prevalence of H. pylori infection and gastric acid secretion in Japanese patients with reflux esophagitis with or without Barrett's esophagus. We enrolled 112 reflux esophagitis patients who were examined for the status of H. pylori and acid secretion in this study. They were divided into three groups, according to the presence or absence of Barrett's esophagus as follows: reflux esophagitis group without Barrett's esophagus (reflux esophagitis alone) (80 patients); short-segment Barrett's esophagus group (16 patients); and long-segment Barrett's esophagus group (LSBE) (16 patients). Age- and sex-matched control subjects were also assigned to the 80 patients with reflux esophagitis alone. The prevalence of H. pylori infection was determined by histology, rapid urease tests, and serum IgG antibodies. Gastric acid secretion was evaluated by the endoscopic gastrin test (EGT). The overall prevalence of H. pylori infection in the reflux esophagitis patient group (24.1%) was significantly lower than the control group (71.2%) (odds ratio 0.13, 95% confidence interval 0.07-0.24; p < 0.0001). The prevalence of H. pylori infection in the patients with Barrett's esophagus tended to be lower than that in the patients with reflux esophagitis alone (reflux esophagitis alone; 30.0%, SSBE; 18.7%, LSBE; 0%), especially in the patients with LSBE compared with the reflux esophagitis alone group (p < 0.01). The EGT value of the respective reflux esophagitis patient group was significantly higher than the control group. The EGT value in the patients with Barrett's esophagus tended to be higher than that in the patients with reflux esophagitis alone, but the difference was not statistically significant. When examined in H. pylori-negative subjects, no difference was found in the EGT value between the control subjects and the patients with reflux esophagitis alone, but it was significantly higher in patients with Barrett's esophagus than the control subjects (p < 0.05). On the other hand, when examined in the H. pylori-positive subjects, the EGT value was significantly higher in the patients with reflux esophagitis alone than in the control subjects (p < 0.01). H. pylori infection may play a protective role in the development of Barrett's esophagus, especially in the development of LSBE in Japan. Gastric acid hypersecretion may be concerned with the development of Barrett's esophagus in addition to the absence of H. pylori infection.


Engineering cytochrome c peroxidase into cytochrome P450: a proximal effect on heme-thiolate ligation.
In an effort to investigate factors required to stabilize heme-thiolate ligation, key structural components necessary to convert cytochrome c peroxidase (CcP) into a thiolate-ligated cytochrome P450-like enzyme have been evaluated and the H175C/D235L CcP double mutant has been engineered. The UV-visible absorption, magnetic circular dichroism (MCD) and electron paramagnetic resonance (EPR) spectra for the double mutant at pH 8.0 are reported herein. The close similarity between the spectra of ferric substrate-bound cytochrome P450cam and those of the exogenous ligand-free ferric state of the double mutant with all three techniques support the conclusion that the latter has a pentacoordinate, high-spin heme with thiolate ligation. Previous efforts to prepare a thiolate-ligated mutant of CcP with the H175C single mutant led to Cys oxidation to cysteic acid [Choudhury et al. (1994) J. Biol. Chem. 267, 25656-25659]. Therefore it is concluded that changing the proximal Asp235 residue to Leu is critical in forming a stable heme-thiolate ligation in the resting state of the enzyme. To further probe the versatility of the CcP double mutant as a ferric P450 model, hexacoordinate low-spin complexes have also been prepared. Addition of the neutral ligand imidazole or of the anionic ligand cyanide results in formation of hexacoordinate adducts that retain thiolate ligation as determined by spectral comparison to the analogous derivatives of ferric P450cam. The stability of these complexes and their similarity to the analogous forms of P450cam illustrates the potential of the H175C/D235L CcP double mutant as a model for ferric P450 enzymes. This study marks the first time a stable cyanoferric complex of a model P450 has been made and demonstrates the importance of the environment around the primary coordination ligands in stabilizing metal-ligand ligation.


Identification of molecular targets associated with ethanol toxicity and implications in drug development.
Alcohol dependence is a major disease burden of adults in modern society worldwide. There is no cure for alcohol dependence. In this study, we have examined the molecular targets of ethanol-induced toxicity in humans based on a systematic review of literature data and then discussed current and potential therapeutic targets for alcohol abuse and dependence. Using human samples with ethanol exposure, microarray analyses of gene expression have shown that numerous genes are up- and/or down-regulated by alcohol exposure. The ethanol-responsive genes mainly encode functional proteins such as proteins involved in nucleic acid binding, transcription factors, selected regulatory molecules, and receptors. These genes are also correlated with important biological pathways, such as angiogenesis, integrin signalling pathway, inflammation, wnt signaling pathway, platelet-derived growth factor signaling pathway, p53 pathway, epidermal growth factor receptor signaling pathway and apoptosis signaling pathway. Currently, only three medications were approved by the U.S. Food and Drug Administration (FDA) for the treatment of alcohol abuse and alcohol dependence, including the aldehyde dehydrogenase inhibitor disulfiram, the micro-opioid receptor antagonist naltrexone, and the N-methyl-D-aspartate (NMDA) receptor inhibitor acamprosate (oral and injectable extended-release formulations). In addition, a number of agents are being investigated as novel treatments for alcohol abuse and dependence. These include selective 5-HT reuptake inhibitors (e.g. fluoxetine), 5-HT(1) receptor agonists (e.g. buspirone), 5-HT(2) receptor antagonists (e.g. ritanserin), 5-HT(3) receptor antagonists (e.g. ondansetron), dopamine receptor antagonists (e.g. aripiprazole and quetiapine), dopamine receptor agonists (e.g. bromocriptine), GABA(B) receptor agonists (e.g. baclofen), and cannabinoid-1 (CB(1)) receptor antagonists. Some of these agents have shown promising efficacy in initial clinical studies. However, further randomized studies with larger samples are warranted to establish their efficacy and safety profiles in the treatment of alcohol dependence.


Prevalence of meconium ileus marks the severity of mutations of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene.
Meconium ileus (MI) is a perinatal complication in cystic fibrosis (CF), which is only minimally influenced by environmental factors. We derived and examined MI prevalence (MIP) scores to assess CFTR phenotype-phenotype correlation for severe mutations. MIP scores were established using a Canadian CF population (n = 2,492) as estimates of the proportion of patients with MI among all patients carrying the same CFTR mutation, focusing on patients with p.F508del as the second allele. Comparisons were made to the registries from the US CF Foundation (n = 43,432), Italy (Veneto/Trentino/Alto Adige regions) (n = 1,788), and Germany (n = 3,596). The prevalence of MI varied among the different registries (13-21%). MI was predominantly prevalent in patients with pancreatic insufficiency carrying "severe" CFTR mutations. In this severe spectrum MIP scores further distinguished between mutation types, for example, G542X (0.31) with a high, F508del (0.22) with a moderate, and G551D (0.08) with a low MIP score. Higher MIP scores were associated with more severe clinical phenotypes, such as a lower forced expiratory volume in 1 second (P = 0.01) and body mass index z score (P = 0.04). MIP scores can be used to rank CFTR mutations according to their clinical severity and provide a means to expand delineation of CF phenotypes.Genet Med 18 4, 333-340.


Lymph node density in muscle-invasive transitional cell carcinoma of the urinary bladder; De novo versus progressive disease.
The prognosis of bladder cancer patients with positive lymph node (LN) disease is affected by both the extent of lymphadenectomy and LN density retrieved during radical cystectomy. This study aimed at assessing the differences in LN metastasis between patients who presented primarily with muscle-invasive transitional cell carcinoma of the bladder "de novo disease" versus "progressive disease." The latter is defined as patients who progressed to muscle-invasive bladder cancer (MIBC) following prior conservative management of a non-muscle-invasive disease. Data were prospectively collected from consecutive 41 radical cystectomies that were divided into two groups: Group I included de novo MIBC cases and Group II included progressive MIBC cases. The median age was 60 years (44-75). Thirty-four patients exhibited de novo disease versus 7 patients who presented as progressive MIBC with a median duration of 9 months between the resection of the first non-invasive tumor and the diagnosis of progressive MIBC (range: 6-56 months). The median number of retrieved LNs in both groups was 15 LNs (range: 4-36). Ten patients (24.39%) had positive pathological LN disease; distributed as 9 patients in Group I and 1 patient in Group II. The median LN density of LN-positive patients was 15.73% (6.46 % in Group I, 28.57% in Group II). Five patients had LN density >20%. Although non-muscle-invasive urothelial bladder tumor may progress to muscle-invasive disease, it still carries less aggressive course than de novo MIBC based on differences in LN metastasis and density.


Continuous paravertebral block for postoperative pain compared to general anaesthesia and wound infiltration for major oncological breast surgery.
We hypothesized that improved acute postoperative pain relief will be achieved using general anaesthesia (GA) either in combination with continuous thoracic paravertebral block (GA-cPVB) or single shot (GA-sPVB) as compared to GA supplemented by local wound infiltration (GA-LWI) after unilateral major breast cancer surgery. A randomised controlled trial was conducted in 46 adult women in a day-care or short-stay hospital setting after major breast cancer surgery. Pain-intensity was measured using an 11-point visual analogue scale (VAS) until postoperative day 2. GA-sPVB was stopped due to slow inclusion. No significant difference in VAS score was noted between GA-LWI (VAS median 0.5 (interquartile range 0.18-2.00)) and GA-cPVB, (VAS 0.3 (0.00-1.55, p = 0.195)) 24 hours after surgery or at any point postoperatively until postoperative day 2. We conclude that both GA-LWI and GA-cPVB anaesthetic techniques are equally effective in treatment of acute postoperative pain after major oncological breast surgery. As GA-LWI is easily to perform with fewer complications and it is more cost-effective it should be preferred over GA-cPVB.


[Temporal bone transverse fracture and peripheric facial paralysis: our surgical approach and results].
In this study, our surgical approaches in temporal bone transvers fracture cases with facial paralysis and their results were evaluated. Five temporal bone transverse fracture cases (2 females, 3 males; mean age 32.8 years; range 4 to 62 years) with facial paralysis which were treated in our clinic between July 1998 and July 2008 were included in this study. Radiologic, audiologic, topographic and electrodiagnostic investigations were made preoperatively. The evaluation of facial nerve function was performed by using House-Brackmann (HB) classification. Tympanic segment, ganglion geniculi and labyrinthine segment were exposed by translabyrinthine approach in all cases. Integrity of the facial nerve was achieved via re-routing and end-to-end anastomosis or n. auricularis major interpositional graft. The cases were evaluated regarding complications and facial nerve function postoperatively. In the audiologic investigation ipsilateral total neurosensorial hearing loss was determined in all cases. In four cases facial paralysis was recognized immediately and in one patient it was recognized when the patient was discharged from the intensive care unit. Facial nerve function was HB stage V in all cases. Surgery was performed in the first six weeks and it was observed that integrity of the labyrinthine segment was distorted in all cases. Reconstruction was performed by re-routing and end-to-end anastomosis in three cases and n. auricularis major interpositional graft in two cases. There were no postoperative complications. Facial nerve function was HB stage II in three cases (re-routing) and HB stage III in two cases (graft). In temporal bone fracture cases with neurosensorial hearing loss and facial paralysis, we achieved good exposure via translabyrintine approach. Better functional results were obtained in re-routing and end-to-end anastomosis technique when compared to interpositional grafting.


Cardiopulmonary bypass parameters and hemostatic response to cardiopulmonary bypass in infants versus children.
Because infants have relatively more blood loss (mL/kg) than older children during cardiac surgery involving cardiopulmonary bypass (CPB), the authors compared hemostatic activation between infants and older children undergoing cardiac surgery. Observational study. University-affiliated children's hospital. Twenty-eight children (18 infants <1 year and 10 children >1 year) undergoing cardiac surgery with CPB. None. Markers of coagulation and fibrinolysis were evaluated at 9 sample points before, during, and after CPB in the 28 children. Infants had greater chest tube output, longer CPB times, and a larger drop in platelet counts during CPB than children. Active tissue plasminogen activator (tPA) increased during CPB in both groups, with infants showing lower levels than children (p < 0.001). In both groups, active plasminogen activator inhibitor type 1 (PAI-1) first decreased during CPB and then increased above baseline postoperatively. Infants had higher PAI-1 than children near the end of CPB (p = 0.01). Thrombin-antithrombin complex levels increased during and after CPB, with infants showing lower levels only during CPB (p = 0.01). D-dimer and prothrombin activation peptide (F1.2) levels increased in a similar pattern for both groups during and after CPB. The length of aortic cross-clamp time and the level of F1.2 after protamine administration correlated significantly and independently with 12-hour chest tube output. Compared with children, infants had greater blood loss (mL/kg), greater drop in platelets during CPB, lower active tPA, and higher active PAI-1. Cumulative thrombin generation after CPB, indicated by F1.2 levels, correlated with early blood loss.


Sustainability of motor performance after robotic-assisted treadmill therapy in children: an open, non-randomized baseline-treatment study.
The aim of the study was to investigate the sustainability of motor improvements achieved after a three week trial of robotic assisted treadmill therapy in children and adolescents with central gait disorders within a follow up period of about six months. Open, non-randomized, baseline-treatment study. Fourteen patients (mean age 8.2+/-5.4) underwent a trial of 12 sessions of robotic-assisted treadmill therapy using the Lokomat over a period of three weeks. Outcome measures were the dimensions D (standing) and E (walking) of the Gross Motor Function Measure, the ten meter walking test and the six minute walking test. Outcome variables were evaluated immediately before and after the trial and at a follow up of about six months. Improvements after the trial in the dimension D from 49.5% to 54.4% (P=0.008) and from 38.9% to 42.3% (P=0.012) in the dimension E of the GMFM were seen and are within the same range of previously published results. The mean score at the follow up after six months was 56.8% and 43.3% for dimension D and E, respectively. Gait speed improved from 0.80 m/s to 1.01 m/s (P=0.006) after the trial and was 1.11 m/s at the follow-up visit at six months. Similar results were obtained for endurance. The improvements of motor function after a three-week trial of robotic-assisted treadmill therapy appear to be sustained after a mean period of six months.


A Petri net modeling approach to explore the temporal dynamics of the provision of multiple ecosystem services.
The representation of the temporal dynamics of ecosystem services (ES) is a crucial research frontier in the field of ES modeling. In fact, most current ES models focus on static ES assessments, that need to be repeated with different inputs per time step to explore potential changes in ES. Here, we present a new approach for the dynamic modeling of multiple ES, based on the Petri Net modeling framework. The key features are: (i) multiple ES are modeled together as a single network, using a social-ecological systems (SES) perspective; (ii) the model accounts for the interactions occurring among ES, by distinguishing between the ES whose provision is mediated by some type of human input, which can produce some side-effects on the system, and those that are generated directly through ecosystem functions and do not generate side-effects; (iii) the model can reproduce the effects of changing drivers on the elements of the SES. These features allow to use the model to explore how ES can evolve over time under different "what-if" scenarios. The importance of considering the ES interactions is tested, showing that failing to include them in the model remarkably affects the results. Due to its complexity, the model should be used as an exploratory tool, focusing on the analysis of the general trends of multiple ES provision, rather than on the generation of quantitative projections. A first conceptual application to the Venice lagoon, Italy, is presented, in which the trends of 13 different ES are simulated. This application shows the potential of the model in exploring the development produced by climate change and socio-economic pressures, and the effects of a set of possible management actions. This modeling approach can contribute to generate new perspectives on the dynamic modeling of multiple ES and on the integrated management of SES.


Psychological Distress and Quality of Life in Pediatric Crohn Disease: Impact of Pain and Disease State.
For patients with Crohn disease (CD), symptom reporting may not coincide with disease state; patients in remission may continue to report symptoms and pain, whereas other patients may be symptom-free despite a flare. This phenomenon has been documented in adults but only recently assessed in pediatric patients. The present study assessed the role of pain reporting and disease state in pediatric patients with CD in understanding psychological distress and quality of life. Participants included 116 children and adolescents ages 8 to 18 years with CD who completed self-report questionnaires assessing pain, disease symptoms, depression, anxiety, functional disability, and quality of life. Physicians completed the Pediatric Crohn's Disease Activity Index to assess disease activity (scores ≤10 = remission, scores >10 = flare). Approximately two thirds of participants reported pain concordant with disease state. For patients in remission, those with pain experienced significantly increased disability and decreased quality of life compared to patients in remission without pain. For patients in a flare, those without pain experienced significantly decreased disability and depressive symptoms, and improved quality of life compared to patients in a flare with pain. For pediatric patients with CD, report of pain, while in remission or a flare, is associated with increased disability and reduced quality of life. Although levels of depression did not differ by disease state, depressive symptoms did differ by pain report (presence or absence) for those in a flare. Pain reporting in CD appears to be associated with both physical and psychological state and should be assessed regardless of disease activity.


The biological effects of long-term exposure of human bronchial epithelial cells to total particulate matter from a candidate modified-risk tobacco product.
Cigarette smoking is the leading cause of preventable lung cancer (LC). Reduction of harmful constituents by heating rather than combusting tobacco may have the potential to reduce the risk of LC. We evaluated functional and molecular changes in human bronchial epithelial BEAS-2B cells following a 12-week exposure to total particulate matter (TPM) from the aerosol of a candidate modified-risk tobacco product (cMRTP) in comparison with those following exposure to TPM from the 3R4F reference cigarette. Endpoints linked to lung carcinogenesis were assessed. Four-week 3R4F TPM exposure resulted in crisis and epithelial to mesenchymal transition (EMT) accompanied by decreased barrier function and disrupted cell-to-cell contacts. By week eight, cells regained E-cadherin expression, suggesting that EMT was reversible. Increased levels of inflammatory mediators were noted in cells treated to 3R4F TPM but not in cells treated to the same or a five-fold higher concentration of cMRTP TPM. A 20-fold higher concentration of cMRTP TPM increased oxidative stress and DNA damage and caused reversible EMT. Anchorage-independent growth was observed in cells treated to 3R4F or a high concentration of cMRTP TPM. 3R4F TPM-derived clones were invasive, while cMRTP TPM-derived clones were not. Long-term exposure to TPM from the cMRTP had a lower biological impact on BEAS-2B cells compared with that of exposure to TPM from 3R4F.


A cGAS-dependent response links DNA damage and senescence in alveolar epithelial cells: a potential drug target in IPF.
Alveolar epithelial cell (AEC) senescence is implicated in the pathogenesis of idiopathic pulmonary fibrosis (IPF). Mitochondrial dysfunction including release of mitochondrial DNA (mtDNA) is a feature of senescence, which led us to investigate the role of the DNA-sensing guanine monophosphate-adenine monophosphate (GMP-AMP) synthase (cGAS) in IPF, with a focus on AEC senescence. cGAS expression in fibrotic tissue from lungs of patients with IPF was detected within cells immunoreactive for epithelial cell adhesion molecule (EpCAM) and p21, epithelial and senescence markers, respectively. Submerged primary cultures of AECs isolated from lung tissue of patients with IPF (IPF-AECs, n = 5) exhibited higher baseline senescence than AECs from control donors (Ctrl-AECs, n = 5-7), as assessed by increased nuclear histone 2AXγ phosphorylation, p21 mRNA, and expression of senescence-associated secretory phenotype (SASP) cytokines. Pharmacological cGAS inhibition using RU.521 diminished IPF-AEC senescence in culture and attenuated induction of Ctrl-AEC senescence following etoposide-induced DNA damage. Short interfering RNA (siRNA) knockdown of cGAS also attenuated etoposide-induced senescence of the AEC line, A549. Higher levels of mtDNA were detected in the cytosol and culture supernatants of primary IPF- and etoposide-treated Ctrl-AECs when compared with Ctrl-AECs at baseline. Furthermore, ectopic mtDNA augmented cGAS-dependent senescence of Ctrl-AECs, whereas DNAse I treatment diminished IPF-AEC senescence. This study provides evidence that a self-DNA-driven, cGAS-dependent response augments AEC senescence, identifying cGAS as a potential therapeutic target for IPF.


Low-dose phase retrieval of biological specimens using cryo-electron ptychography.
Cryo-electron microscopy is an essential tool for high-resolution structural studies of biological systems. This method relies on the use of phase contrast imaging at high defocus to improve information transfer at low spatial frequencies at the expense of higher spatial frequencies. Here we demonstrate that electron ptychography can recover the phase of the specimen with continuous information transfer across a wide range of the spatial frequency spectrum, with improved transfer at lower spatial frequencies, and as such is more efficient for phase recovery than conventional phase contrast imaging. We further show that the method can be used to study frozen-hydrated specimens of rotavirus double-layered particles and HIV-1 virus-like particles under low-dose conditions (5.7 e/Å2) and heterogeneous objects in an Adenovirus-infected cell over large fields of view (1.14 × 1.14 μm), thus making it suitable for studies of many biologically important structures.


Diet selection by Japanese quail (Coturnix coturnix japonica) in relation to ambient temperature and metabolic rate.
1. A choice between a high-energy, wheat-based, low protein mixture and a lower-energy, soya-based, high protein mixture offered to growing Japanese quail at ambient temperatures of 20 degrees, 25 degrees, 30 degrees and 35 degrees C. 2. the quail were kept in open-circuit respiration calorimeters, so that diet selection could be related to energy requirement. 3. Increasing ambient temperature had no significant effect on food intake by weight, but the proportion of the high energy choice decreased and, conversely, the proportion of the lower-energy but higher-protein choice increased. 4. Energy intake was therefore negatively correlated with ambient temperature, but protein intake per unit of energy intake increased, allowing the birds to gain weight at about the same rate at all temperatures. 5. Heat production decreased as ambient temperature increased. Respiratory quotient decreased with increasing temperature, which indicates a reduced utilisation of carbohydrate as an energy source. 6. Water intake increased with temperature but there were no overt signs of heat stress and there was no significant change in body temperature. 7. Japanese quail selected a dietary mixture which maintained similar growth rates over a wide range of ambient temperature, by sustaining protein intake but altering energy intake in line with thermoregulatory energy demands.


Singlet-triplet conversion and the long-range proximity effect in superconductor-ferromagnet structures with generic spin dependent fields.
The long-range proximity effect in superconductor-ferromagnet (S/F) hybrid nanostructures is observed if singlet Cooper pairs from the superconductor are converted into triplet pairs which can diffuse into the ferromagnet over large distances. It is commonly believed that this happens only in the presence of magnetic inhomogeneities. We show that there are other sources of the long-range triplet component (LRTC) of the condensate and establish general conditions for their occurrence. As a prototypical example, we consider first a system where the exchange field and spin-orbit coupling can be treated as time and space components of an effective SU(2) potential. We derive a SU(2) covariant diffusive equation for the condensate and demonstrate that an effective SU(2) electric field is responsible for the long-range proximity effect. Finally, we extend our analysis to a generic ferromagnet and establish a universal condition for the LRTC. Our results open a new avenue in the search for such correlations in S/F structures and make a hitherto unknown connection between the LRTC and Yang-Mills electrostatics.


Adolescent dual diagnosis: a 12-step transformational model.
A structured model inpatient treatment program for multiple-diagnosed treatment-resistant adolescents is described. The chemical dependence disease model of Predisposition+Drug+Enabling System=Disease provides the fundamental theoretical approach. This model is elaborated and used similarly for psychiatric disorders, with examples presented. The role of psychiatric evaluation for pharmacological interventions is outlined. The psychotherapeutic treatment focuses on a commitment to abstinence as the goal. A "psychology of commitment" is described using a reframing of traditional psychotherapeutic approaches. This approach is used in conjunction with the first five steps of Alcoholics Anonymous as an empowering therapeutic model. The role of defenses, denial, confrontation, and cognitive distortions is described. The value of this approach as a neutral ground for discussion using concepts from traditional Freudian therapy, self-psychology, cognitive and learning therapies, and rational emotive therapy is elaborated.


Design strategies and rigorous conditions for single-polarization single-mode waveguides.
We establish rigorous necessary analytical conditions for the existence of single-polarization single-mode (SPSM) bandwidths in index-guided microstructured waveguides (such as photonic-crystal fibers). These conditions allow us to categorize designs for SPSM waveguides into four strategies, at least one of which seems previously unexplored. Conversely, we obtain rigorous sufficient conditions for the existence of two cutoff-free index-guided modes in a wide variety of microstructured dielectric waveguides with arbitrary periodic claddings, based on the existence of a degenerate fundamental mode of the cladding (a degenerate light line). We show how such a degenerate light line, in turn, follows from the symmetry of the cladding.


Effects of a stability ball exercise programme on low back pain and daily life interference during pregnancy.
most pregnant women experience back pain during pregnancy, a serious issue that negatively impacts life quality during pregnancy. Research into an exercise intervention programme targeting low back pain and daily life interference is lacking. this study evaluates how a stability ball exercise programme influences low back pain and daily life interference across the second and third pregnancy trimester. the study was non-randomised and controlled, examining a target population of low-risk pregnancy women between 20 and 22 weeks of gestation located in a regional hospital in northern Taiwan. All participants had at least minimal low back pain, no prior history of chronic low back pain before pregnancy, and no indications of preterm labour. In total, 89 individuals participated: 45 in the control group and 44 in the experimental group (who attended an antenatal stability ball exercise programme). This programme lasted 12 weeks, composed of at least three sessions per week. Fitness workouts lasted from 25 to 30 minutes. The women completed their basic personal information, the Brief Pain Inventory-Short Form, and the Family Exercise Support Attitude Questionnaire. after adjusting for demographic data and antenatal exercise status by propensity scores, experimental-group women who participated in the antenatal stability ball exercise programme reported significantly less low back pain and daily life interferences than the control group at 36 weeks of gestation. the inclusion of stability ball exercises during pregnancy may reduce pregnancy low back pain and boost daily life functions. This stability ball exercise programme provides health-care professionals with an evidence-based intervention.


Mental Health Outcomes Among Homeless, Runaway, and Stably Housed Youth.
Runaway youth and homeless youth are at risk for adverse mental health outcomes. These 2 populations are frequently pooled together in both research and interventions yet may have unique health needs. We sought to assess differences in mental health outcomes among these populations. We conducted a secondary data analysis of ninth- and 11th-graders in the 2016 minnesota Student Survey (n = 68 785). We categorized youth into 4 subgroups based on housing status in the previous year: (1) unaccompanied homeless youth (0.5%), (2) runaway youth (4%), (3) youth who had both run away and been homeless (0.6%), and (4) stably housed youth (95%). We performed multivariable logistic regression to compare 4 mental health outcomes (self-injury, suicidal ideation, suicide attempts, and depressive symptoms) across groups, controlling for demographics and abuse history. Unstably housed youth had poorer mental health outcomes when compared with their stably housed peers (P < .05). For example, 11% of homeless youth, 20% of runaways, and 33% of youth who had experienced both had attempted suicide in the previous year compared with 2% of stably housed youth (adjusted odds ratios 2.4, 4.9, and 7.1, respectively). Other outcomes showed a similar pattern. Our findings suggest that runaway and homeless youth represent unique populations with high levels of mental health needs who would benefit from targeted clinical and community interventions. Pediatric clinicians represent one potential point of screening and intervention.


Glutamate mediates a slow synaptic response in hippocampal slice cultures.
Glutamate (GLU) mediates its 'fast' excitatory transmitter action in the brain by directly gating cation-selective ion channels ('ionotropic' receptors). However, GLU can also activate another type of receptor, coupled to phospholipase C ('metabotropic' receptor). In hippocampal cells, stimulation of this metabotropic receptor by GLU, or by a racemic mixture of (1S-3R and 1R-3S) 1-aminocyclopentyl-1,3-dicarboxylate (ACPD), induces a slower excitation mediated by inhibition of K+ currents. We have assessed whether this slow form of metabotropic receptor excitation can contribute to the effects of synaptically released GLU in hippocampal slice cultures, by recording the responses of CA3 pyramidal cells to afferent mossy fibre stimulation. When the fast ionotropic response was blocked pharmacologically, mossy fibre stimulation produced a slow depolarizing postsynaptic potential associated with a decrease in membrane conductance, a depression of the slow after-hyperpolarization following a train of action potentials, and reduced accommodation during the action potential train. Under voltage-clamp, mossy fibre stimulation produced a slow voltage-dependent inward current which resembled that produced by application of exogenous ACPD or quisqualate (QUIS), and which was occluded by these metabotropic agonists. We therefore suggest that synaptically released GLU can induce two types of postsynaptic responses: a fast excitation through activation of ionotropic receptors and a slower excitation associated with inhibition of K+ conductances through activation of metabotropic receptors. This is analogous to the dual action of acetylcholine on ionotropic (nicotinic) and metabotropic (muscarinic) receptors.


Advice that includes food sources of unsaturated fat supports future risk management of gestational diabetes mellitus.
Abstract Women with gestational diabetes mellitus (GDM) have a greater risk of developing type 2 diabetes mellitus (DM) and heart disease than pregnant women without GDM. Advice given during the GDM pregnancy provides an opportunity to develop protective dietary patterns for the long-term management of this risk. Dietary guidelines for the prevention and management of type 2 DM support the inclusion of unsaturated fats, but food advice needs to target this outcome. The aim of this study was to compare the dietary intakes of women with GDM given general low-fat advice (control group) to women with GDM given the same advice with additional targets for food sources of unsaturated fats (intervention group). After approximately 6 weeks, the intervention group reported more ideal dietary fatty acid intakes than the control group, with polyunsaturated:saturated fat ratios of 1:1 and 0.4:1, respectively ( P < .001), assessed using repeated measures analysis of variance. These results confirm the need to include food sources of unsaturated fats in advice strategies to assure optimal protective eating habits in this at-risk group.


Assortative mating, sexual selection, and their consequences for gene flow in Littorina.
When divergent populations are connected by gene flow, the establishment of complete reproductive isolation usually requires the joint action of multiple barrier effects. One example where multiple barrier effects are coupled consists of a single trait that is under divergent natural selection and also mediates assortative mating. Such multiple-effect traits can strongly reduce gene flow. However, there are few cases where patterns of assortative mating have been described quantitatively and their impact on gene flow has been determined. Two ecotypes of the coastal marine snail, Littorina saxatilis, occur in North Atlantic rocky-shore habitats dominated by either crab predation or wave action. There is evidence for divergent natural selection acting on size, and size-assortative mating has previously been documented. Here, we analyze the mating pattern in L. saxatilis with respect to size in intensively sampled transects across boundaries between the habitats. We show that the mating pattern is mostly conserved between ecotypes and that it generates both assortment and directional sexual selection for small male size. Using simulations, we show that the mating pattern can contribute to reproductive isolation between ecotypes but the barrier to gene flow is likely strengthened more by sexual selection than by assortment.


Membrane insertion of the N-terminal alpha-helix of equinatoxin II, a sea anemone cytolytic toxin.
Equinatoxin II (Eqt-II) is a member of the actinoporins, a unique family of cytotoxins comprising 20 kDa pore-forming proteins isolated from sea anemones. Actinoporins bind preferentially to lipid membranes containing sphingomyelin, and create cation-selective pores by oligomerization of three to four monomers. Previous studies have shown that regions of Eqt-II crucial for its cytolytic mechanism are an exposed aromatic cluster and the N-terminal region containing an amphipathic alpha-helix. In the present study, we have investigated the transfer of the N-terminal alpha-helix into the lipid membrane by the use of three mutants containing an additional tryptophan residue in different positions within the amphipathic alpha-helix (Ile18-->Trp, Val22-->Trp and Ala25-->Trp). The interaction of the mutants with different model systems, such as lipid monolayers, erythrocytes and ghost membranes, was extensively characterized. Intrinsic fluorescence measurements and the use of vesicles containing brominated phospholipids indicated a deep localization of the N-terminal amphipathic helix in the lipid bilayer, except for the case of Val22-->Trp. This mutant is stabilized in a state immediately prior to final pore formation. The introduction of additional tryptophan residues in the sequence of Eqt-II has proved to be a suitable approach to monitor the new environments that surround defined regions of the molecule upon membrane interaction.


A good surgical option for ischemic mitral regurgitation in co-morbid patients: semicircular reduction annuloplasty.
Ring annuloplasty is the standard treatment of ischemic mitral regurgitation (MR), however, it has been associated with some drawbacks. It abolishes normal annular dynamics and freezes the posterior leaflet. In the present study, we evaluated Paneth suture annuloplasty in chronic ischemic MR and both early and mid-term outcomes of the technique on a selected population. The study period was from June 2010 to June 2012. We operated on 21 patients who had the diagnosis of coronary artery disease and MR of grade 3 or 4. The patients had both a coronary artery bypass operation and the mitral semicircular reduction annuloplasty described by Paneth-Burr. The data on the patients were retrospectively collected. Patients were contacted by outpatient clinic controls for mid-term results. The male/female ratio was 10/11. The mean age of the patients was 71.0 ± 6.4 years. Preoperative and postoperative left ventricular ejection fraction was statistically similar (P = 0.973). Early postoperative MR grade (mean, 0.57 ± 0.51) was statistically lower than the preoperative MR grades (mean, 3.38 ± 0.50) (P < 0.001). There was no revision for excess bleeding. Two patients had prolonged hospitalization, one for sternal infection and the other for severe chronic obstructive pulmonary disease. No hospital or late postoperative deaths occurred. The mean late postoperative MR grade was 0.66 ± 0.97 degrees. One patient had progression of MR in the later follow-up, which was treated by mitral valve replacement. Semicircular reduction annuloplasty is an effective, inexpensive and easy surgical annuloplasty technique with low mortality and morbidity in severe symptomatic ischemic MR.


A blinded randomised controlled trial to determine the effect of enteric coating on enzyme treatment for canine exocrine pancreatic efficiency.
Enzyme treatment is the mainstay for management of exocrine pancreatic insufficiency (EPI) in dogs. 'Enteric-coated' preparations have been developed to protect the enzyme from degradation in the stomach, but their efficacy has not been critically evaluated. The hypothesis of the current study was that enteric coating would have no effect on the efficacy of pancreatic enzyme treatment for dogs with EPI.Thirty-eight client-owned dogs with naturally occurring EPI were included in this multicentre, blinded, randomised controlled trial. Dogs received either an enteric-coated enzyme preparation (test treatment) or an identical preparation without the enteric coating (control treatment) over a period of 56 days. There were no significant differences in either signalment or cobalamin status (where cobalamin deficient or not) between the dogs on the test and control treatments. Body weight and body condition score increased in both groups during the trial (P<0.001) but the magnitude of increase was greater for the test treatment compared with the control treatment (P<0.001). By day 56, mean body weight increase was 17% (95% confidence interval 11-23%) in the test treatment group and 9% (95% confidence interval 4-15%) in the control treatment group. The dose of enzyme required increased over time (P<0.001) but there was no significant difference between treatments at any time point (P=0.225). Clinical disease severity score decreased over time for both groups (P=0.011) and no difference was noted between groups (P=0.869). No significant adverse effects were reported, for either treatment, for the duration of the trial. Enteric coating a pancreatic enzyme treatment improves response in canine EPI.


Phthalate diesters in airborne PM(2.5) and PM(10) in a suburban area of Shanghai: seasonal distribution and risk assessment.
Concentrations of nine phthalate diesters in 24-h airborne PM2.5 and PM10 were determined from October 2011 to August 2012 in a suburban area in Shanghai, China. Dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), di-iso-butyl phthalate (DIBP), benzyl butyl phthalate (BzBP), and di(2-ethylhexyl) phthalate (DEHP) were frequently detected in airborne particulate matter at sum concentrations of these six compounds ranging from 13.3 to 186 ng/m(3), with an average value of 59.8 ng/m(3) in PM2.5, and from 10.1 to 445 ng/m(3), with an average value of 132 ng/m(3) in PM10. DEHP, DBP, and DIBP were the major phthalate diesters found in PM samples. DEHP was found predominantly in coarse (size fraction of between PM2.5 and PM10) particles, whereas DMP, DEP, DBP, DIBP, and BzBP were found predominantly in fine (PM2.5) particles. The concentrations of phthalates in PM during warm months (207 ng/m(3) for PM10 and 71.9 ng/m(3) for PM2.5, on average) were significantly higher than those during cold months (76.9 ng/m(3) for PM10 and 50.4 ng/m(3) for PM2.5). Significant positive correlations were found between concentrations of total phthalates, DEHP, and BzBP, with the total mass and organic carbon content of PM. Based on the concentrations of DEHP, incremental lifetime cancer risks (ILCR) from inhalation exposure were estimated using a Monte Carlo simulation. Although the 95% probabilities for the ILCR values for the general population were below the U.S. Environmental Protection Agency (EPA) threshold of 10(-6), our result is an underestimate of the actual health risk because we only considered the outdoor inhalation exposure to DEHP in this study.


Assessment of the Prognostic Value of Radiomic Features in 18F-FMISO PET Imaging of Hypoxia in Postsurgery Brain Cancer Patients: Secondary Analysis of Imaging Data from a Single-Center Study and the Multicenter ACRIN 6684 Trial.
Hypoxia is associated with resistance to radiotherapy and chemotherapy in malignant gliomas, and it can be imaged by positron emission tomography with 18F-fluoromisonidazole (18F-FMISO). Previous results for patients with brain cancer imaged with 18F-FMISO at a single center before conventional chemoradiotherapy showed that tumor uptake via T/Bmax (tissue SUVmax/blood SUV) and hypoxic volume (HV) was associated with poor survival. However, in a multicenter clinical trial (ACRIN 6684), traditional uptake parameters were not found to be prognostically significant, but tumor SUVpeak did predict survival at 1 year. The present analysis considered both study cohorts to reconcile key differences and examine the potential utility of adding radiomic features as prognostic variables for outcome prediction on the combined cohort of 72 patients with brain cancer (30 University of Washington and 42 ACRIN 6684). We used both 18F-FMISO intensity metrics (T/Bmax, HV, SUV, SUVmax, SUVpeak) and assessed radiomic measures that determined first-order (histogram), second-order, and higher-order radiomic features of 18F-FMISO uptake distributions. A multivariate model was developed that included age, HV, and the intensity of 18F-FMISO uptake. HV and SUVpeak were both independent predictors of outcome for the combined data set (P < .001) and were also found significant in multivariate prognostic models (P < .002 and P < .001, respectively). Further model selection that included radiomic features showed the additional prognostic value for overall survival of specific higher order texture features, leading to an increase in relative risk prediction performance by a further 5%, when added to the multivariate clinical model..


The Family Check-up Intervention Moderates Polygenic Influences on Long-Term Alcohol Outcomes: Results from a Randomized Intervention Trial.
Alcohol problems are influenced by both genetic and environmental factors. Evidence from twin models and measured gene-environment interaction studies has demonstrated that the importance of genetic influences changes as a function of the environment. Research has also shown that family-centered interventions may protect genetically susceptible youth from developing substance use problems. In this study, we brought large-scale gene identification findings into an intervention study to examine gene-by-intervention effects. Using genome-wide polygenic scores derived from an independent genome-wide association study of adult alcohol dependence, we examined whether an adolescent family-centered intervention would moderate the effect of genetic risk for alcohol dependence on lifetime alcohol dependence in young adulthood, approximately 15 years after the start of intervention, among European American (N = 271; 48.3% in the intervention condition) and African American individuals (N = 192; 51.6% in the intervention condition). We found that among European American individuals, the intervention moderated the association between alcohol dependence polygenic scores and lifetime alcohol dependence diagnosis in young adulthood. Among participants in the control condition, higher alcohol dependence polygenic scores were associated with a greater likelihood of receiving an alcohol dependence diagnosis; in contrast, among participants in the intervention condition, there was no association between alcohol dependence polygenic scores and alcohol dependence diagnosis. No moderation effect was found among African Americans. These results demonstrate that modifying environments of genetically vulnerable youth could reduce the likelihood of developing alcohol dependence and underscore the significance of environmentally focused prevention and intervention efforts.


Change in Perceived Stress and Health Behaviors of Emerging Adults in the Midst of the COVID-19 Pandemic.
The COVID-19 pandemic has resulted in unprecedented disruptions, restrictions, and concerns about physical and mental health. Emerging adulthood, including the first year of college, is associated with declines in healthy eating and physical activity, as well as possible heightened distress. The impact of COVID-19 may exacerbate these concerns. The purpose of this study was to examine changes in health behaviors and perceived stress in emerging adults over the first year of college and to determine whether prepandemic health behaviors were protective for mental health and stress during the initial changes after the COVID-19 pandemic. First-year college students (N = 234, 58.6% female) completed three surveys during their first year of school, the third being after the onset of COVID-19 and during a stay-at-home order. At Time 3, we also assessed symptoms of anxiety and depression. Using linear mixed modeling, sedentary time increased and physical activity decreased over time, but 20%-35% of students reported improvements in these behaviors. Dietary changes appeared mixed, with some improvements noted early during COVID-19. Perceived stress increased over time. Multiple regression indicated that of the health behaviors examined for protective effects on mental health and stress during the pandemic, only diet quality emerged as a significant predictor. Although notable declines in some health habits were observed over time, including following COVID-19 disruptions, some students reported improved health behaviors. Efforts should be directed at identifying and intervening with students most at risk for poor functioning.


Giant abdominal wall abscess dissecting into thorax as a complication of ESWL.
Abscess as a complication of extracorporeal shock wave lithotripsy is a rare condition. We present the computed tomography findings of an abdominal wall abscess that occurred after extracorporeal shock wave lithotripsy for which prophylactic antibiotics had not been given. The abscess destroyed the posterior abdominal wall muscles and dissected into the thorax. The muscles were thickened and showed enhancement. A parenchymal defect in the right kidney adjacent to a caliceal stone, with strands extending from this defective region to the abscess, was observed, and was thought to be the result of parenchymal destruction caused by the shock waves. Other parts of the kidney and psoas muscle were normal. Microbiologic examination revealed Escherichia coli.


Candesartan cilexetil loaded solid lipid nanoparticles for oral delivery: characterization, pharmacokinetic and pharmacodynamic evaluation.
Candesartan cilexetil (CC) is used in the treatment of hypertension and heart failure. It has poor aqueous solubility and low oral bioavailability. In this work, CC loaded solid lipid nanoparticles (CC-SLNs) were developed to improve the oral bioavailability. Components of the SLNs include either of trimyristin/tripalmitin/tristearin, and surfactants (Poloxamer 188 and egg lecithin E80). The CC loaded nanoparticles were prepared by hot homogenization followed by ultrasonication method. The physicochemical properties, morphology of CC-SLNs were characterized, the pharmacokinetic and pharmacodynamic behaviour of CC-SLNs were evaluated in rats. Stable CC-SLNs having a mean particle size of 180-220 nm with entrapment efficiency varying in between 91-96% were developed. The physical stability of optimized formulation was studied at refrigerated and room temperature for 3 months. Further, freeze drying was tried for improving the physical stability. DSC and XRD analyses indicated that the drug incorporated into SLN was in amorphous form but not in crystalline state. The SLN-morphology was found to be nearly spherical by electron microscopic studies. Pharmacokinetic results indicated that the oral bioavailability of CC was improved over 2.75-fold after incorporation into SLNs. Pharmacodynamic study of SLNs in hypertensive rats showed a decrease in systolic blood pressure for 48 h, while suspension showed a decrease in systolic blood pressure for only 2 h. Taken together, these effects are due to enhanced bioavailability coupled with sustained action of CC in SLN formulation. Thus, the results conclusively demonstrated the role of CC-SLNs for a significant enhancement in oral bioavailability along with improved pharmacodynamic effect.


Behavior of an extradural pressure monitor in clinical use. Comparison of extradural with intraventricular pressure in patients with acute and chronically raised intracranial pressure.
A comparison of intraventricular pressure (IVP) and extradural pressure (EDP) was carried out in 17 patients being investigated for normal-pressure hydrocephalus, and in six patients with acutely raised intracranial pressure following events such as head injury or intracerebral hematoma. Extradural pressure was measured using the CardioSearch monitor. There was a reasonably good correlation between EDP and IVP in the chronic stable group with pressures up to 25 mm Hg. In the acute group there was no predictable relationship between EDP and IVP, and during a 24-hour period the pressures could vary by as much as 30 mm Hg. Subdural pressure, measured with the same instruments, was compared to IVP in both acute and stable situations in eight other patients: there was a close and constant correlation between pressures in these two spaces. The authors conclude that misleading information may be obtained from EDP monitoring, and erroneous management decisions may result from dependence on such a technique. Possible explanations for this are discussed.


LEDGF/p75 determines cellular trafficking of diverse lentiviral but not murine oncoretroviral integrase proteins and is a component of functional lentiviral preintegration complexes.
Human immunodeficiency virus type 1 (HIV-1), feline immunodeficiency virus (FIV), and Moloney murine leukemia virus (MoMLV) integrases were stably expressed to determine their intracellular trafficking. Each lentiviral integrase localized to cell nuclei in close association with chromatin while the murine oncoretroviral integrase was cytoplasmic. Fusions of pyruvate kinase to the lentiviral integrases did not reveal transferable nuclear localization signals. The intracellular trafficking of each was determined instead by the transcriptional coactivator LEDGF/p75, which was required for nuclear localization. Stable small interfering RNA expression eliminated detectable LEDGF/p75 expression and caused dramatic, stable redistribution of each lentiviral integrase from nucleus to cytoplasm while the distribution of MoMLV integrase was unaffected. In addition, endogenous LEDGF/p75 coimmunoprecipitated specifically with each lentiviral integrase. In vitro integration assays with preintegration complexes (PICs) showed that endogenous LEDGF/p75 is a component of functional HIV-1 and FIV PICs. However, HIV-1 and FIV infection and replication in LEDGF/p75-deficient cells was equivalent to that in control cells, whether cells were dividing or growth arrested. Two-long terminal repeat circle accumulation in nondividing cell nuclei was also equivalent to that of LEDGF/p75 wild-type cells. Virions produced in LEDGF/p75-deficient cells had normal infectivity. We conclude that LEDGF/p75 fully accounts for cellular trafficking of diverse lentiviral, but not oncoretroviral, integrases and is the main lentiviral integrase-to-chromatin tethering factor. While lentiviral PIC nuclear import is unaffected by LEDGF/p75 knockdown, this protein is a component of functional lentiviral PICs. A role in HIV-1 integration site distribution merits investigation.


The yolk stalk sign: evidence of death in small embryos without heartbeats.
The purpose of this study was to assess the positive predictive value for confirming early embryonic death in the clinical sonographic scenario wherein an embryo is identified without a visible heartbeat; the embryonic crown-rump length (CRL) is 5 mm or less; and the embryo is not immediately adjacent to the yolk sac. A retrospective study of 882 first-trimester sonograms was performed among women who had an intrauterine pregnancy of uncertain viability based on 1 or more sonographic findings (eg, no visible heartbeat in an embryo with a CRL of < or =5 mm). Eight hundred six cases met the inclusion criteria. Among the cohort of 806 cases, 520 (64.5%) had an identifiable embryo. One hundred fifty-nine of these embryos had no demonstrable heartbeat and a CRL of 5 mm or less. The CRLs of these embryos ranged from 1.7 to 5.4 mm. This cohort's sonograms were reviewed to determine whether there was a separation between the embryo and yolk sac. Twenty-one cases were discovered. Recall that as a retrospective study, no specific effort was made to show this finding. Thus, a computation of the sensitivity of this finding would result in an underestimate of indeterminate magnitude. All of these cases were subsequently proven to be failed pregnancies. The positive predictive value of the "yolk stalk sign" in determining early pregnancy failure for an embryo with a CRL of 5 mm or less and no visible heartbeat was 100% in this cohort.


Association between subarachnoid hemorrhage outcomes and number of cases treated at California hospitals.
Studies of several complex medical conditions have shown that outcomes are better at hospitals that treat more cases. We tested the hypothesis that patients with subarachnoid hemorrhage treated at high-volume hospitals have better outcomes. Using a database of all admissions to nonfederal hospitals in California from 1990 to 1999, we obtained discharge abstracts for patients with a primary diagnosis of subarachnoid hemorrhage who were admitted through the emergency department. Hospital volume, defined as the average number of subarachnoid hemorrhage cases admitted each year, was divided into quartiles. Rates of mortality, adverse outcomes (death or discharge to long-term care), length of stay, and hospital charges were computed by univariate analysis and by multivariable general estimating equations, with adjustment for demographic and admission characteristics. A total of 12 804 patients were admitted for subarachnoid hemorrhage through the emergency departments of 390 hospitals. Hospital volumes varied from 0 to 8 cases per year in the first quartile to 19 to 70 cases per year in the fourth quartile. The mortality rate in the lowest volume quartile (49%) was larger than that in the highest volume quartile (32%, P<0.001). In multivariable analysis, the difference persisted (odds ratio comparing highest with lowest volume quartiles 0.57, 95% CI 0.48 to 0.67; P<0.001). At higher volume hospitals, lengths of stay were longer, and hospital charges were greater in univariate and multivariable models (all P<0.001). Only 4.8% of those admitted to hospitals in the lowest volume quartile were transferred to hospitals in the highest quartile. In this study of discharge abstracts in California, hospitals that treated more cases of subarachnoid hemorrhage had substantially lower rates of in-hospital mortality. Few patients with subarachnoid hemorrhage are being transferred to high-volume centers.


The aggregation-enhancing huntingtin N-terminus is helical in amyloid fibrils.
The 17-residue N-terminus (htt(NT)) directly flanking the polyQ sequence in huntingtin (htt) N-terminal fragments plays a crucial role in initiating and accelerating the aggregation process that is associated with Huntington's disease pathogenesis. Here we report on magic-angle-spinning solid-state NMR studies of the amyloid-like aggregates of an htt N-terminal fragment. We find that the polyQ portion of this peptide exists in a rigid, dehydrated amyloid core that is structurally similar to simpler polyQ fibrils and may contain antiparallel β-sheets. In contrast, the htt(NT) sequence in the aggregates is composed in part of a well-defined helix, which likely also exists in early oligomeric aggregates. Further NMR experiments demonstrate that the N-terminal helical segment displays increased dynamics and water exposure. Given its specific contribution to the initiation, rate, and mechanism of fibril formation, the helical nature of htt(NT) and its apparent lack of effect on the polyQ fibril core structure seem surprising. The results provide new details about these disease-associated aggregates and also provide a clear example of an amino acid sequence that greatly enhances the rate of amyloid formation while itself not taking part in the amyloid structure. There is an interesting mechanistic analogy to recent reports pointing out the early-stage contributions of transient intermolecular helix-helix interactions in the aggregation behavior of various other amyloid fibrils.


3 W, 300 μJ, 25 ns pulsed 473 nm blue laser based on actively Q-switched Nd:YAG single-crystal fiber oscillator at 946 nm.
We report the realization of a frequency doubled, actively Q-switched and polarized oscillator based on Nd:YAG single-crystal fiber. A laser output of 8 W, 10 kHz, and 30 ns at 946 nm is reported. The laser is extracavity frequency doubled in a BiBO crystal to obtain 3 W and 300 μJ of blue laser with a beam quality of M(2)y=1.12 and M(2)x=1.38. The obtained blue power is stable with a root-mean-square stability of less than 2% in 1 h. This is more than two times the previously reported average power and energy at 473 nm.


[Evaluation of leptin levels in plasma and their reliance on other hormonal factors affecting tissue fat levels in people with various levels of endogenous cotisol].
The discovery of leptin (LEP) shed new light on mechanisms regulating body fat mass (BFM). In this aspect, interactions between LEP and glucocorticoids at hypothalamic level may be of great importance. Factors that influence plasma LEP levels have not been fully recognized and available data on LEP levels are often inconsistent. The aim of this study was to evaluate absolute and BFM-corrected plasma LEP levels and their diurnal variation, as well as to assess the relationship between LEP levels, body fat distribution, and hormones influencing body fat in subjects with various levels of endogenous cortisol and different nutritional status. Group I was composed of 14 women aged 14-58 yrs, BMI of 23.9-37.1 kg/m2, with hypercortisolism due to ACTH-dependent and ACTH-independent Cushing's syndrome (CUS). 17 women with visceral obesity (OTY) and normal or disturbed carbohydrate metabolism, i.e. impaired glucose tolerance (IGT) and diabetes mellitus (DM), aged 24 do 50 yrs, BMI 30.0-46.1 kg/m2, were included in group II. Group III consisted of 14 women with Addison's disease (AD), aged 18 do 63 yrs, BMI 15.4-31.6 kg/m2. The control group IV (KON) included 17 healthy women with normal BMI. BMI, WHR, body composition, and body fat distribution (DEXA method) were assessed in all subjects. Basal plasma levels of LEP, beta-endorphin (B-EP), cortisol (F), insulin-like growth factor-1 (IGF-1) were measured with RIA test kits. Plasma adrenocorticotrophin (ACTH) levels, serum levels of insulin (IRI) and growth hormone (GH) were measured with IRMA test kits. Blood glucose (G) concentration was determined with an enzymatic method. Adiposity-corrected LEP levels were expressed as LEP/BFM and LEP/%BF indices. Fasting insulin resistance index (FIRI) was also calculated. Higher BFM and %BF values were found in the OTY group as compared with CUS KON and AD groups. BFM distribution did not differ in KON and AD groups whereas CUS subjects exhibited a higher accumulation of fat in the trunk when compared to OTY subjects. Absolute LEP levels were correlated with trunk BF in CUS patients whereas in KON and AD groups these levels were correlated only with limb fat. Absolute LEP levels in CUS and OTY groups were comparable, whereas LEP/BFM and LEP/%BF indices were higher in the CUS group (Table 1) reflecting upregulation of LEP levels (Figs. 1, 2). BFM-corrected LEP levels were comparable in groups with normal cortisolemia, i.e. in OTY and KON groups, whereas in the AD group both absolute and BFM-corrected LEP levels were lower than in controls. No correlation was found between plasma levels of F and LEP in CUS and AD groups. This correlation was negative in KON (Fig. 3) and positive in OTY groups (Fig. 4). Moreover, KON and AD groups demonstrated a negative correlation between plasma ACTH and LEP levels. CUS patients showed positive, BFM-independent correlations between LEP levels, FIRI and G values, and a positive, BFM-dependent correlation between IRI and LEP levels. OTY patients exhibited a BFM-dependent positive correlation between FIRI and LEP levels. In these and in AD patients, a positive, BFM-independent correlation between IRI and LEP levels was found. Moreover, a negative, BFM-dependent correlation between GH and LEP levels was found in OTY patients. In this group, B-EP levels were positively correlated with LEP/BFM and LEP/%BF indices (Fig. 5). A negative correlation between LEP levels, LEP/BFM and LEP/%BF indices was ascertained in the AD group. In CUS, OTY, and KON groups, but not in the AD group, a midnight increase in leptin levels was observed. In conclusion, upregulation of leptin levels in relation to body fat in Cushing's syndrome is independent of the source of hypercortisolism. Apparently, it results from insulin resistance and hyperglycaemia and contributes to coexisting metabolic abnormalities. In Addison's disease, downregulation of leptin may reflect an adaptation mechanism to cortisol deficiency and result from low insulin and extremely high adrenocorticotrophin levels. In women with normal cortisol levels, irrespectively of nutritional status; leptin levels reflect body fat content. In obese subjects, leptin levels may be influenced by cortisol levels, high levels of insulin, IGF-1, and beta-endorphin as well as low levels of growth hormone. Disturbed function of hypothalamic-pituitary-adrenal axis (CUS, AD) does not directly influence diurnal variation in plasma leptin levels. In Cushing's syndrome, visceral fat may be a predominant source of leptin, whereas in women with normal or low cortisol levels peripherally accumulated fat may determine leptin secretion.


Agonist-driven conformational changes in the inner beta-sheet of alpha7 nicotinic receptors.
Cys-loop ligand-gated ion channels assemble as pentameric proteins, and each monomer contributes two structural elements: an extracellular ligand-binding domain (LBD) and a transmembrane ion channel domain. Models of receptor activation include rotational movements of subunits leading to opening of the ion channel. We tested this idea using substituted cysteine accessibility to track conformational changes in the inner beta sheet of the LBD. Using a nondesensitizing chick alpha7 background (L(247)T), we constructed 18 consecutive cysteine replacement mutants (Leu(36) to Ile(53)) and tested each for expression of acetylcholine (ACh)-evoked currents and functional sensitivity to thiol modification. We measured rates of modification in the presence and absence of ACh to identify conformational changes associated with receptor activation. Resting modification rates of eight substituted cysteines in the beta1 and beta2 strands and the sequence between them (loop 2) varied over several orders of magnitude, suggesting substantial differences in the accessibility or electrostatic environment of individual side chains. These differences were in general agreement with structural models of the LBD. Eight of 18 cysteine replacements displayed ACh-dependent changes in modification rates, indicating a change in the accessibility or electrostatic environment of the introduced cysteine during activation. We were surprised that the effects of agonist exposure were difficult to reconcile with rotational models of activation. Acetylcholine reduced the modification rate of M(40)C but increased it at N(52)C despite the close physical proximity of these residues. Our results suggest that models that depend strictly on rigid-body rotation of the LBD may provide an incomplete description of receptor activation.


[Endoscopic diagnosis of local chemical burn of mucous membranes of the stomach, induced with the purpose of simulation of gastric ulcer].
With the purpose of improvement of diagnosis of induced gastric ulcer were examined 11 patients who took aggressive agents for simulation of gastric ulcer and 33 patients who took pseudo-aggressive agents. Observables, conduced diagnosis of local chemical burn of mucous coat of stomach during initial 6 days after taking aggressive agents. Stages of ulcerous process, resulting from local chemical burn of mucous coat of stomach, coressponds to real gactric ulcer. Gelatin capsule using as a container for delivery of aggressive agents, melts in stomach in 5-6 minutes after taking. Independent from body position, mucous coat of greater curvature of the stomach is damaged. It is impossible to simulate duodenal bulb ulcer using the gelatine capsule or ball made of breadcrumb. The last method of delivery of aggressive agent can damage the small intestine because of uncontrollability of the place of breaking the ball.


Simultaneous Prediction of four ATP-binding Cassette Transporters' Substrates Using Multi-label QSAR.
Efflux by the ATP-binding cassette (ABC) transporters affects the pharmacokinetic profile of drugs and it has been implicated in drug-drug interactions as well as its major role in multi-drug resistance in cancer. It is therefore important for the pharmaceutical industry to be able to understand what phenomena rule ABC substrate recognition. Considering a high degree of substrate overlap between various members of ABC transporter family, it is advantageous to employ a multi-label classification approach where predictions made for one transporter can be used for modeling of the other ABC transporters. Here, we present decision tree-based QSAR classification models able to simultaneously predict substrates and non-substrates for BCRP1, P-gp/MDR1 and MRP1 and MRP2, using a dataset of 1493 compounds. To this end, two multi-label classification QSAR modelling approaches were adopted: Binary Relevance (BR) and Classifier Chain (CC). Even though both multi-label models yielded similar predictive performances in terms of overall accuracies (close to 70 %), the CC model overcame the problem of skewed performance towards identifying substrates compared with non-substrates, which is a common problem in the literature. The models were thoroughly validated by using external testing, applicability domain and activity cliffs characterization. In conclusion, a multi-label classification approach is an appropriate alternative for the prediction of ABC efflux.


Mechanism for leptin's acute insulin-independent effect to reverse diabetic ketoacidosis.
The mechanism by which leptin reverses diabetic ketoacidosis (DKA) is unknown. We examined the acute insulin-independent effects of leptin replacement therapy in a streptozotocin-induced rat model of DKA. Leptin infusion reduced rates of lipolysis, hepatic glucose production (HGP), and hepatic ketogenesis by 50% within 6 hours and were independent of any changes in plasma glucagon concentrations; these effects were abrogated by coinfusion of corticosterone. Treating leptin- and corticosterone-infused rats with an adipose triglyceride lipase inhibitor blocked corticosterone-induced increases in plasma glucose concentrations and rates of HGP and ketogenesis. Similarly, adrenalectomized type 1 diabetic (T1D) rats exhibited decreased rates of lipolysis, HGP, and ketogenesis; these effects were reversed by corticosterone infusion. Leptin-induced decreases in lipolysis, HGP, and ketogenesis in DKA were also nullified by relatively small increases (15 to 70 pM) in plasma insulin concentrations. In contrast, the chronic glucose-lowering effect of leptin in a STZ-induced mouse model of poorly controlled T1D was associated with decreased food intake, reduced plasma glucagon and corticosterone concentrations, and decreased ectopic lipid (triacylglycerol/diacylglycerol) content in liver and muscle. Collectively, these studies demonstrate marked differences in the acute insulin-independent effects by which leptin reverses fasting hyperglycemia and ketoacidosis in a rodent model of DKA versus the chronic pleotropic effects by which leptin reverses hyperglycemia in a non-DKA rodent model of T1D.


The L1624Q Variant in SCN1A Causes Familial Epilepsy Through a Mixed Gain and Loss of Channel Function.
Variants of the SCN1A gene encoding the neuronal voltage-gated sodium channel NaV1.1 cause over 85% of all cases of Dravet syndrome, a severe and often pharmacoresistent epileptic encephalopathy with mostly infantile onset. But with the increased availability of genetic testing for patients with epilepsy, variants in SCN1A have now also been described in a range of other epilepsy phenotypes. The vast majority of these epilepsy-associated variants are de novo, and most are either nonsense variants that truncate the channel or missense variants that are presumed to cause loss of channel function. However, biophysical analysis has revealed a significant subset of missense mutations that result in increased excitability, further complicating approaches to precision pharmacotherapy for patients with SCN1A variants and epilepsy. We describe clinical and biophysical data of a familial SCN1A variant encoding the NaV1.1 L1624Q mutant. This substitution is located on the extracellular linker between S3 and S4 of Domain IV of NaV1.1 and is a rare case of a familial SCN1A variant causing an autosomal dominant frontal lobe epilepsy. We expressed wild-type (WT) and L1642Q channels in CHO cells. Using patch-clamp to characterize channel properties at several temperatures, we show that the L1624Q variant increases persistent current, accelerates fast inactivation onset and decreases current density. While SCN1A-associated epilepsy is typically considered a loss-of-function disease, our results put L1624Q into a growing set of mixed gain and loss-of-function variants in SCN1A responsible for epilepsy.


Polymer-Based Nitric Oxide Therapies: Recent Insights for Biomedical Applications.
Since the discovery of nitric oxide (NO) in the 1980s, this cellular messenger has been shown to participate in diverse biological processes such as cardiovascular homeostasis, immune response, wound healing, bone metabolism, and neurotransmission. Its beneficial effects have prompted increased research in the past two decades, with a focus on the development of materials that can locally release NO. However, significant limitations arise when applying these materials to biomedical applications. This Feature Article focuses on the development of NO-releasing and NO-generating polymeric materials (2006-2011) with emphasis on recent in vivo applications. Results are compared and discussed in terms of NO dose, release kinetics, and biological effects, in order to provide a foundation to design and evaluate new NO therapies.


Pleural effusion in childhood falciparum malaria.
Pulmonary complication is a rare manifestation of childhood malaria and isolated pleural effusion without pulmonary edema has never been reported in children. We report here an 11-year-old boy who suffered from cerebral malaria and massive right pleural effusion. The patient was treated with intravenous artesunate, albumin, and other supportive treatments. He recovered completely after eight days. The clinical and laboratory courses suggested that the plasma leakage played a role in the pathogenesis of pleural effusion.