Novel SERS-based process analysis for label-free segmented flow screenings.
In microfluidic segmented flow processes label-free analytical techniques like surface enhanced Raman spectroscopy (SERS) can reveal the chemical composition of the individual droplet contents. The SERS system developed in this work enables a simple connection to micro segmented flow processes through miniaturization. The concept is based on the parallelization of silver/polyacrylamide composite SERS spots on a carrier plate on which the segments are deposited. The transfer of the segments allows an easy connection to existing flow processes and provides optimal conditions for Raman measurements using miniaturized spectrometers. The preparation of the SERS polymer composite was optimized in terms of the silver content in the polymer matrix to obtain a high SERS signal. The performance and long-term stability of the polymer have been successfully demonstrated. The deamination of adenine with sodium nitrite to hypoxanthine was chosen as a case study to demonstrate the capability of the novel SERS-based process analysis. A sequence of approximately one hundred segments in combination with a gradient of the nitrite concentration (0 to 0.4 mol L-1) was generated at two temperatures. The concentration of adenine and hypoxanthine were determined by using a multivariate calibration model, since the Raman spectra of both substances are overlapping. It was shown that the conversion of adenine is increased with higher nitrite concentration and temperature. A conversion of 35% was obtained at 50 °C and a conversion of 60% at 80 °C, respectively.


[Spongy cardiomyopathy in an elderly woman. Echocardiographic description].
Isolated left ventricular noncompaction, also known as spongy myocardium or spongy cardiomyopathy, is a recently described congenital disease caused by an arrest in the left ventricular myocardial embriogenesis that makes the ventricular wall to persist thickened with multiple trabecular formations and deep sinusoidal recesses. It is clinically characterized by heart failure, cardiac arrhythmia and systemic embolic events. Most of the affected subjects are detected during childhood or adolescence, others in the adult life but very few elderly patients have been reported in the worldwide medical literature. We here report the case of a 75-year-old woman that is one of the oldest patients ever reported, whose clinical picture and echocardiographic findings are typical of this modality of cardiomyopathy. We do comments on this case in regard to the most relevant facts that appear in the limited medical literature about this interesting disease.


Expression of TMEFF2 in Human Pancreatic Cancer Tissue and the Effects of TMEFF2 Knockdown on Cell, Proliferation, and Apoptosis in Human Pancreatic Cell Lines.
BACKGROUND The TMEFF2 gene encodes the transmembrane protein with EGF like and two follistatin-like domains 2 and has been reported to be a tumor suppressor gene, but its role remains unknown in pancreatic cancer. This study aimed to investigate the expression of TMEFF2 in human pancreatic cancer tissue and the effects of knockdown of TMEFF2 on cell, proliferation, and apoptosis in human pancreatic cell lines. MATERIAL AND METHODS Thirty-five samples of human pancreatic tissue and adjacent normal pancreatic tissue, and five human pancreatic cancer cell lines, CAPAN1, ASPC1, BXPC3, SW1990, and CFPAC were studied. RNA expression, protein expression, cell proliferation, and apoptosis were studied using real-time polymerase chain reaction (RT-PCR), Western blot, the cell counting kit-8 (CCK-8) assay, and flow cytometry, respectively. A co-immunoprecipitation assay evaluated protein interactions. RESULTS TMEFF2 expression was down-regulated in pancreatic cancer tissue compared with normal pancreas. In human pancreatic cancer cell lines, overexpression of TMEFF2 suppressed cell proliferation and enhanced apoptosis, suppressed the expression of p-STAT3, MCL1, VEGF and increased the expression of the tyrosine-specific protein phosphatase, SHP-1. The co-immunoprecipitation assay showed that TMEFF2 interacted with SHP-1. Knockdown of expression of TMEFF2 resulted in the increased expression of p-STAT3, MCL1, and VEGF, increased cell proliferation and decreased cell apoptosis, which were reversed by overexpression of SHP-1. CONCLUSIONS In pancreatic cancer, TMEFF2 exerted as a tumor suppressor effect by regulating p-STAT3, MCL1, and VEGF via SHP-1.


Discovery and antitumor evaluation of novel inhibitors of spermine oxidase.
Increasing knowledge of the relationship between cancer and dysregulated polyamine catabolism suggests interfering with aberrant polyamine metabolism for anticancer therapy that will have considerable clinical promise. SMO (spermine oxidase) plays an essential role in regulating the polyamines homeostasis. Therefore, development of SMO inhibitors has increasingly attracted much attention. Previously, we successfully purified and characterised SMO. Here, we presented an in silico drug discovery pipeline by combining pharmacophore modelling and molecular docking for the virtual screening of SMO inhibitors. In vitro evaluation showed that N-(3-{[3-(dimethylamino)propyl]amino}propyl)-8-quinolinecarboxamide (SI-4650) inhibited SMO enzyme activity, increased substrate spermine content and reduced product spermidine content, indicating that SI-4650 can interfere with polyamine metabolism. Furthermore, SI-4650 treatment suppressed cell proliferation and migration. Mechanistically, SI-4650 caused cell cycle arrest, induced cell apoptosis, and promoted autophagy. These results demonstrated the properties of interfering with polyamine metabolism of SI-4650 as a SMO inhibitor and the potential for cancer treatment.


Use of tazarotene foam for the treatment of acne vulgaris.
Acne vulgaris is a common inflammatory chronic disease of the pilosebaceous unit. It often requires long-term treatment, resulting in increased demand for topical medications that are popular with patients in order to achieve long-term compliance. Tazarotene foam 0.1% is a novel formulation of tazarotene. We review efficacy and tolerability studies of the new formulation, and suggest a possible place for the product in the management of acne vulgaris.


Estrogen action in the regulation of cell proliferation, cell survival, and tumorigenesis in the rat anterior pituitary gland.
Estrogens act as important regulators of cell proliferation, cell survival, and differentiation in a variety of organ systems and tissues and have been implicated in the etiology of a variety of malignant cancers and benign tumors. The anterior pituitary gland of the rat provides an excellent model for the study of estrogen action in the regulation of cell proliferation and survival. Estrogens stimulate proliferation of the prolactin (PRL)-producing lactotroph and enhance lactotroph survival. Through these actions on lactotroph proliferation and survival, estrogens induce or contribute to the development of PRL-producing pituitary tumors in several rat strains. Data from our laboratory and others indicate that estrogen-induced pituitary growth is rat strain specific and segregates as a quantitative genetic trait in crosses between different rat strains. The purpose of this review is to summarize current knowledge pertaining to estrogen action in the regulation of cell proliferation, cell survival, and tumorigenesis in the anterior pituitary gland of the rat species, Rattus norvegicus, and to illustrate the advantages of the rat pituitary gland as a model for elucidating the mechanisms through which estrogens regulate these processes.


Machine Learning Prediction of Crossbred Pig Feed Efficiency and Growth Rate From Single Nucleotide Polymorphisms.
This research assessed the ability of a Support Vector Machine (SVM) regression model to predict pig crossbred (CB) performance from various sources of phenotypic and genotypic information for improving crossbreeding performance at reduced genotyping cost. Data consisted of average daily gain (ADG) and residual feed intake (RFI) records and genotypes of 5,708 purebred (PB) boars and 5,007 CB pigs. Prediction models were fitted using individual PB genotypes and phenotypes (trn.1); genotypes of PB sires and average of CB records per PB sire (trn.2); and individual CB genotypes and phenotypes (trn.3). The average of CB offspring records was the trait to be predicted from PB sire's genotype using cross-validation. Single nucleotide polymorphisms (SNPs) were ranked based on the Spearman Rank correlation with the trait. Subsets with an increasing number (from 50 to 2,000) of the most informative SNPs were used as predictor variables in SVM. Prediction performance was the median of the Spearman correlation (SC, interquartile range in brackets) between observed and predicted phenotypes in the testing set. The best predictive performances were obtained when sire phenotypic information was included in trn.1 (0.22 [0.03] for RFI with SVM and 250 SNPs, and 0.12 [0.05] for ADG with SVM and 500-1,000 SNPs) or when trn.3 was used (0.29 [0.16] with Genomic best linear unbiased prediction (GBLUP) for RFI, and 0.15 [0.09] for ADG with just 50 SNPs). Animals from the last two generations were assigned to the testing set and remaining animals to the training set. Individual's PB own phenotype and genotype improved the prediction ability of CB offspring of young animals for ADG but not for RFI. The highest SC was 0.34 [0.21] and 0.36 [0.22] for RFI and ADG, respectively, with SVM and 50 SNPs. Predictive performance using CB data for training leads to a SC of 0.34 [0.19] with GBLUP and 0.28 [0.18] with SVM and 250 SNPs for RFI and 0.34 [0.15] with SVM and 500 SNPs for ADG. Results suggest that PB candidates could be evaluated for CB performance with SVM and low-density SNP chip panels after collecting their own RFI or ADG performances or even earlier, after being genotyped using a reference population of CB animals.


Child abuse and neglect.
Child abuse and neglect are important public health problems and recent estimates of their prevalence suggest that they are considerably more common than had hitherto been realized. Intervening to change parenting practices may, however, be important in their treatment. Despite their frequent occurrence among dental patients, neglect is the least known and identified type of abuse. The present case reports a 3-year-old girl suffering from abuse and neglect. The girl had dental neglect and also performed poorly in all parameters of growth and development. This case describes the process that was followed in order to reach a diagnosis and safeguard the child. The parent was counseled and the girl got her due share of access to comprehensive oral and general health. This case emphasizes that the dental professional should always be vigilant and collaborates among like-minded individuals to increase the prevention, detection and treatment of these conditions.


Resting energy expenditure in obese women: comparison between measured and estimated values.
Assessing energy requirements is a fundamental activity in clinical dietetic practice. The aim of this study was to investigate which resting energy expenditure (REE) predictive equations are the best alternatives to indirect calorimetry before and after an interdisciplinary therapy in Brazilian obese women. In all, twelve equations based on weight, height, sex, age, fat-free mass and fat mass were tested. REE was measured by indirect calorimetry. The interdisciplinary therapy consisted of nutritional, physical exercise, psychological and physiotherapy support during the course of 1 year. The average differences between measured and predicted REE, as well as the accuracy at the ±10 % level, were evaluated. Statistical analysis included paired t tests, intraclass correlation coefficients and Bland-Altman plots. Validation was based on forty obese women (BMI 30-39·9 kg/m2). Our major findings demonstrated a wide variation in the accuracy of REE predictive equations before and after weight loss in non-morbid, obese women. The equations reported by Harris-Benedict and FAO/WHO/United Nations University (UNU) were the only ones that did not show significant differences compared with indirect calorimetry and presented a bias <5 %. The Harris-Benedict equation provided 40 and 47·5 % accurate predictions before and after therapy, respectively. The FAO equation provided 35 and 47·5 % accurate predictions. However, the Bland-Altman analysis did not show good agreement between these equations and indirect calorimetry. Therefore, the Harris-Benedict and FAO/WHO/UNU equations should be used with caution for obese women. The need to critically re-assess REE data and generate regional and more homogeneous REE databases for the target population is reinforced.


Aerosol Gas-Phase Components from Cannabis E-Cigarettes and Dabbing: Mechanistic Insight and Quantitative Risk Analysis.
Consumption of cannabis by nontraditional methods has surged since the advent of legalization in North America and worldwide. Inhaling cannabis extracts using vaporizers and via dabbing has risen in popularity, while concerns over product safety have not hindered their proliferation. The work herein is the first step toward assessing the safety of vaporizing and dabbing concentrated cannabis extracts as a function of gas-phase reaction products. The gas-phase thermal degradants of Δ9-tetrahydrocannabinol (THC) have not been previously investigated. It was found that users may be exposed to concerning degradants such as methacrolein, benzene, and methyl vinyl ketone when using cartridge vaporizers and dabbing. It was shown that THC alone and mixed with terpenes generated similar degradation products and, most notably, elevated levels of isoprene. Importantly, it was shown that added terpenes led to higher levels of gas-phase products compared to THC alone. To estimate cancer and noncancer risks associated with exposure to these and other degradants, quantitative risk assessment was applied to experimentally determined values for dabbing and vaping and literature-sourced levels of hazardous components in cannabis smoke. Overall, gas-phase aerosol products had significantly lower values in dabbing and vaporizing compared to cannabis smoking, although these results should be interpreted in light of potential variations in degradant levels due to disparate usage patterns and the dangers of the higher aerosol concentration of THC.


Differential gene expression profiles of scirrhous gastric cancer cells with high metastatic potential to peritoneum or lymph nodes.
Scirrhous gastric cancer is often accompanied by metastasis to the peritoneum and/or lymph nodes, resulting in the highest mortality rate among gastric cancers. Mechanisms involved in gastric cancer metastasis are not fully clarified because metastasis involves multiple steps and requires the accumulation of altered expression of many different genes. Thus, independent analysis of any single gene would be insufficient to understand all of the aspects of gastric cancer metastasis. In this study, we performed global analysis on differential gene expression of a scirrhous gastric cancer cell line (OCUM-2M) and its derivative sublines with high potential for metastasis to the peritoneal cavity (OCUM-2MD3) and lymph nodes (OCUM-2MLN) in a nude mice model. By applying a high-density oligonucleotide array method, expression of approximately 6800 genes was analyzed, and selected genes were confirmed by the Northern blot method. In our observations in OCUM-2MD3 cells, 12 genes were up-regulated, and 20 genes were down-regulated. In OCUM-2MLN cells, five genes were up-regulated, and five genes were down-regulated. The analysis revealed two functional gene clusters with altered expression: (a) down-regulation of a cluster of squamous cell differentiation marker genes such as small proline-rich proteins [SPRRs (SPRR1A, SPRR1B, and SPRR2A], annexin A1, epithelial membrane protein 1, cellular retinoic acid-binding protein 2, and mesothelin in OCUM-2MD3 cells; and (b) up-regulation of a cluster of antigen-presenting genes such as MHC class II (DP, DR, and DM) and invariant chain (II) in OCUM-2MLN cells through up-regulation of CIITA (MHC class II transactivator). We then analyzed six gastric cancer cell lines by Northern blot and observed preferential up-regulation of trefoil factor 1, alpha-1-antitrypsin, and galectin 4 and down-regulation of cytidine deaminase in cells prone to peritoneal dissemination. Genes highly correlated with invasion or peritoneal dissemination of gastric cancer, such as E-cadherin or integrin beta4, were down-regulated in both of the derivative cell lines analyzed in this study. This is the first demonstration of global gene expression analysis of gastric cancer cells with different metastatic potentials, and these results provide a new insight in the study of human gastric cancer metastasis.


Brain injury biomarkers may improve the predictive power of the IMPACT outcome calculator.
Outcome prediction following severe traumatic brain injury (sTBI) is a widely investigated field of research. A major breakthrough is represented by the IMPACT prognostic calculator based on admission data of more than 8500 patients. A growing body of scientific evidence has shown that clinically meaningful biomarkers, including glial fibrillary acidic protein (GFAP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), and αII-spectrin breakdown product (SBDP145), could also contribute to outcome prediction. The present study was initiated to assess whether the addition of biomarkers to the IMPACT prognostic calculator could improve its predictive power. Forty-five sTBI patients (GCS score≤8) from four different sites were investigated. We utilized the core model of the IMPACT calculator (age, GCS motor score, and reaction of pupils), and measured the level of GFAP, UCH-L1, and SBDP145 in serum and cerebrospinal fluid (CSF). The forecast and actual 6-month outcomes were compared by logistic regression analysis. The results of the core model itself, as well as serum values of GFAP and CSF levels of SBDP145, showed a significant correlation with the 6-month mortality using a univariate analysis. In the core model, the Nagelkerke R(2) value was 0.214. With multivariate analysis we were able to increase this predictive power with one additional biomarker (GFAP in CSF) to R(2)=0.476, while the application of three biomarker levels (GFAP in CSF, GFAP in serum, and SBDP145 in CSF) increased the Nagelkerke R(2) to 0.700. Our preliminary results underline the importance of biomarkers in outcome prediction, and encourage further investigation to expand the predictive power of contemporary outcome calculators and prognostic models in TBI.


In vivo activity of murine de novo methyltransferases, Dnmt3a and Dnmt3b.
The putative de novo methyltransferases, Dnmt3a and Dnmt3b, were reported to have weak methyltransferase activity in methylating the 3' long terminal repeat of Moloney murine leukemia virus in vitro. The activity of these enzymes was evaluated in vivo, using a stable episomal system that employs plasmids as targets for DNA methylation in human cells. De novo methylation of a subset of the CpG sites on the stable episomes is detected in human cells overexpressing the murine Dnmt3a or Dnmt3b1 protein. This de novo methylation activity is abolished when the cysteine in the P-C motif, which is the catalytic site of cytosine methyltransferases, is replaced by a serine. The pattern of methylation on the episome is nonrandom, and different regions of the episome are methylated to different extents. Furthermore, Dnmt3a also methylates the sequence methylated by Dnmt3a on the stable episome in the corresponding chromosomal target. Overexpression of human DNMT1 or murine Dnmt3b does not lead to the same pattern or degree of de novo methylation on the episome as overexpression of murine Dnmt3a. This finding suggests that these three enzymes may have different targets or requirements, despite the fact that weak de novo methyltransferase activity has been demonstrated in vitro for all three enzymes. It is also noteworthy that both Dnmt3a and Dnmt3b proteins coat the metaphase chromosomes while displaying a more uniform pattern in the nucleus. This is the first evidence that Dnmt3a and Dnmt3b have de novo methyltransferase function in vivo and the first indication that the Dnmt3a and Dnmt3b proteins may have preferred target sites.


Automated Opt-Out vs Opt-In Patient Outreach Strategies for Breast Cancer Screening: A Randomized Clinical Trial.
Optimal strategies for population-based outreach for breast cancer screening remain unknown. To evaluate the effect on breast cancer screening of an opt-out automatic mammography referral strategy compared with an opt-in automated telephone message strategy. This pragmatic randomized clinical trial was conducted from April 2022 to January 2023 at a single Veterans Affairs (VA) medical center. Participants were female veterans aged 45 to 75 years who were eligible for breast cancer screening and enrolled in VA primary care. Veterans were randomized 1:1 to receive either an automatic mammography referral (opt-out arm) or an automated telephone call with an option for mammography referral (opt-in arm). The primary outcome was completed mammography 100 days after outreach. Secondary outcomes were scheduled or completed mammography by 100 days after outreach and referrals canceled if mammography was not scheduled within 90 days. Both intention-to-treat analyses and a restricted analysis were conducted. The restricted analysis excluded veterans who were unable to be reached by telephone (eg, a nonworking number) or who were found to be ineligible after randomization (eg, medical record documentation of recent mammography). Of 883 veterans due for mammography (mean [SD] age, 59.13 [8.24] years; 656 [74.3%] had received prior mammography), 442 were randomized to the opt-in group and 441 to the opt-out group. In the intention-to-treat analysis, there was no significant difference in the primary outcome of completed mammography at 100 days between the opt-out and opt-in groups (67 [15.2%] vs 66 [14.9%]; P = .90) or the secondary outcome of completed or scheduled mammography (84 [19%] vs 106 [24.0%]; P = .07). A higher number of referrals were canceled in the opt-out group compared with the opt-in group (104 [23.6%] vs 24 [5.4%]; P < .001). The restricted analysis demonstrated similar results except more veterans completed or scheduled mammography within 100 days in the opt-out group compared with the opt-in group (102 of 388 [26.3%] vs 80 of 415 [19.3%]; P = .02). In this randomized clinical trial, an opt-out population-based breast cancer screening outreach approach compared with an opt-in approach did not result in a significant difference in mammography completion but did lead to substantially more canceled mammography referrals, increasing staff burden. ClinicalTrials.gov Identifier: NCT05313737.


Serological markers predict inflammatory bowel disease years before the diagnosis.
Anti-neutrophil cytoplasmic antibodies and anti-Saccharomyces cerevisiae mannan antibodies (ASCAs) have been detected in the serum of patients with ulcerative colitis (UC) and Crohn's disease (CD) and their unaffected family members. The aim of this study was to establish the value of serological markers as predictors of UC and CD. Individuals who developed CD or UC were identified from the European Prospective Investigation into Cancer and Nutrition (EPIC) study. At recruitment, none of the participants had a diagnosis of CD or UC. For each incident case, two controls were randomly selected matched for centre, date of birth, sex, date of recruitment and time of follow-up. Serum of cases and controls obtained at recruitment were analysed for ASCA IgG, ASCA IgA, perinuclear anti-neutrophil cytoplasmic antibody (pANCA), antibodies against Escherichia coli outer membrane porin C (OmpC) and flagellin CBir1. Conditional logistic regression was used to determine risk of CD and UC. Receiver operating characteristic curves were constructed to test accuracy. A total of 77 individuals were diagnosed with CD and 167 with UC after a mean follow-up of 4.5 (SD 3.2) and 4.4 (SD 3.1) years following blood collection, respectively. Combinations of pANCA, ASCA, anti-CBir1 and anti-OmpC were most accurate in predicting incident CD and UC (area under curve 0.679 and 0.657, respectively). The predictive value of the combination of markers increased when time to diagnosis of CD or UC decreased. A panel of serological markers is able to predict development of CD and UC in individuals from a low-risk population.


Preservation of hatchery waste by lactic acid fermentation. 1. Laboratory scale fermentation.
In Experiment 1, two types of hatchery wastes, including cockerel chicks (CC), and shell waste (SW) blended with CC (60:40 CC:SW), were ground and mixed with a by-product carbohydrate (5, 10, and 15% wt/wt) with or without a bacterial culture and fermented for a period of 21 d. Cockerel chicks fermented with 15% carbohydrate and the culture had a pleasant smell and little H2S production. Elevated H2S concentrations were recorded for CC:SW samples at all carbohydrate levels when the culture was not added. No NH3 was detected from any treatments during fermentation. The addition of culture to the CC and CC:SW by-products resulted in pH values lower (P < 0.05) than those without culture on Day 21, and the 15% carbohydrate treatment significantly reduced pH beyond the 5% carbohydrate. Final proximate composition of CC and CC:SW samples with culture were not significantly different from those without culture added. In Experiment 2, carbohydrate was added at 10.0, 13.3, 16.7, and 20.0% wt/wt to CC and CC:SW in the presence of the bacterial culture. Shell waste alone was fermented with 15, 20, and 25% carbohydrate and the culture. Moisture level in this experiment was adjusted to approximately 70% for all treatments. The lowest pH for the CC and CC:SW treatments was observed at the 16.7% carbohydrate level. Shell waste pH was better maintained at the 20 and 25% carbohydrate levels. After fermentation for 21 d CC, CC:SW and SW treatments from Experiments 1 and 2 contained negligible Escherichia coli, and no Salmonella were detected.


Acknowledging the impact of seasonal blood pressure variation in hypertensive CKD and non-CKD patients living in a Mediterranean climate.
This study aims to assess seasonal blood pressure (BP) variation in chronic kidney disease (CKD) and non-CKD patients living in a Mediterranean climate, and to find out if this variation entails significant adjustment of treatment and if it impacts renal outcomes and mortality. This retrospective study included all hypertensive patients seen between February 2006 and April 2020 in two Lebanese clinics. Regression analyses were used to assess the association of seasonal BP variability and treatment adjustment with eGFR change from baseline, dialysis initiation and death. A total of 398 patients of 64.2 ±13.9 years were followed for 51.1 ±44.3 months, 67% had eGFR< 60 mL/min. Mean systolic and diastolic BP was 137.7 ±14.7 and 76.5 ±9.5 mmHg respectively. Systolic and diastolic BP were significantly lower in the warm season in CKD and non-CKD patients (P<0.001). The majority (91.4%) needed seasonal treatment modifications. After adjustment to age, sex, baseline eGFR, BP and number of antihypertensive drugs, we found a significant loss of eGFR with treatment modifications in both seasons, double risk of dialysis with the increase of antihypertensive treatment in both seasons and a 2.5 more risk of death with reduced treatment in the warm season. This study confirmed the seasonal BP variability in CKD and non-CKD patients from a Mediterranean climate. All types of treatment adjustment were associated with eGFR loss. Low BP in the warm season was highly associated with death.


Limited Variation between SARS-CoV-2-Infected Individuals in Domain Specificity and Relative Potency of the Antibody Response against the Spike Glycoprotein.
The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is arranged as a trimer on the virus surface, composed of three S1 and three S2 subunits. Infected and vaccinated individuals generate antibodies against spike, which can neutralize the virus. Most antibodies target the receptor-binding domain (RBD) and N-terminal domain (NTD) of S1; however, antibodies against other regions of spike have also been isolated. The interhost variability in domain specificity and relative neutralization efficacy of the antibodies is still poorly characterized. To this end, we tested serum and plasma samples collected from 85 coronavirus disease 2019 (COVID-19) convalescent subjects. Samples were analyzed using seven immunoassays that employ different domains, subunits, and oligomeric forms of spike to capture the antibodies. Samples were also tested for their neutralization of pseudovirus containing SARS-CoV-2 spike and of replication-competent SARS-CoV-2. While the total amount of anti-spike antibodies produced varied among convalescent subjects, we observed an unexpectedly fixed ratio of RBD- to NTD-targeting antibodies. The relative potency of the response (defined as the measured neutralization efficacy relative to the total level of spike-targeting antibodies) also exhibited limited variation between subjects and was not associated with the overall amount of antispike antibodies produced. These studies suggest that host-to-host variation in the polyclonal response elicited against SARS-CoV-2 spike in early pandemic subjects is primarily limited to the quantity of antibodies generated rather than their domain specificity or relative neutralization potency. IMPORTANCE Infection by SARS-CoV-2 elicits antibodies against various domains of the spike protein, including the RBD and NTD of subunit S1 and against subunit S2. The antibody responses of different infected individuals exhibit different efficacies to inactivate (neutralize) the virus. Here, we show that the observed variation in the neutralizing activity of the antibody responses in COVID-19 convalescent subjects is caused by differences in the amounts of antibodies rather than their recognition properties or the potency of their antiviral activity. These findings suggest that COVID-19 vaccine strategies that focus on enhancing the overall level of the antibodies will likely elicit a more uniformly efficacious protective response.


Integrating health belief model and technology acceptance model: an investigation of health-related internet use.
Today, people use the Internet to satisfy health-related information and communication needs. In Malaysia, Internet use for health management has become increasingly significant due to the increase in the incidence of chronic diseases, in particular among urban women and their desire to stay healthy. Past studies adopted the Technology Acceptance Model (TAM) and Health Belief Model (HBM) independently to explain Internet use for health-related purposes. Although both the TAM and HBM have their own merits, independently they lack the ability to explain the cognition and the related mechanism in which individuals use the Internet for health purposes. This study aimed to examine the influence of perceived health risk and health consciousness on health-related Internet use based on the HBM. Drawing on the TAM, it also tested the mediating effects of perceived usefulness of the Internet for health information and attitude toward Internet use for health purposes for the relationship between health-related factors, namely perceived health risk and health consciousness on health-related Internet use. Data obtained for the current study were collected using purposive sampling; the sample consisted of women in Malaysia who had Internet access. The partial least squares structural equation modeling method was used to test the research hypotheses developed. Perceived health risk (β=.135, t1999=2.676) and health consciousness (β=.447, t1999=9.168) had a positive influence on health-related Internet use. Moreover, perceived usefulness of the Internet and attitude toward Internet use for health-related purposes partially mediated the influence of health consciousness on health-related Internet use (β=.025, t1999=3.234), whereas the effect of perceived health risk on health-related Internet use was fully mediated by perceived usefulness of the Internet and attitude (β=.029, t1999=3.609). These results suggest the central role of perceived usefulness of the Internet and attitude toward Internet use for health purposes for women who were health conscious and who perceived their health to be at risk. The integrated model proposed and tested in this study shows that the HBM, when combined with the TAM, is able to predict Internet use for health purposes. For women who subjectively evaluate their health as vulnerable to diseases and are concerned about their health, cognition beliefs in and positive affective feelings about the Internet come into play in determining the use of health-related Internet use. Furthermore, this study shows that engaging in health-related Internet use is a proactive behavior rather than a reactive behavior, suggesting that TAM dimensions have a significant mediating role in Internet health management.


First trimester diagnosis of beta-thalassaemia in a twin pregnancy.
This paper reports the results of first trimester prenatal diagnosis in a twin pregnancy at risk for homozygous beta 0-thalassaemia (beta 0-39 mutant). Trophoblast samples from both twins were obtained at 10 weeks gestation with a forceps guided by ultrasound. Trophoblast DNA analysis, carried out with the oligonucleotide technique, revealed that one fetus was homozygous and the other heterozygous for the beta-39 mutant. This diagnosis was confirmed at 17 weeks gestation by amniocyte DNA analysis. DNA polymorphism analysis within the alpha-globin gene provided useful genetic markers for twin differentiation.


An elderly woman with increasing dyspnoea after a fall.
An otherwise healthy 92-year-old woman was admitted to our department with shortness of breath and dysphagia 10 h after a fall in her bathroom. Medical checkup at another institution had not uncovered the causation of the complaints. Clinical and radiological examinations at our department then revealed an expanding retropharyngeal and prevertebral haematoma. Because of increasing dyspnoea, a lateral cervical approach was used to remove the haematoma and to achieve haemostasis. The authors could demonstrate that the source of bleeding was a minor injury of the anterior longitudinal ligament. Retropharyngeal haematoma is a potentially life-threatening condition because it can rapidly progress to airway obstruction. Large retropharyngeal haematoma after minor blunt head and neck trauma is not a well-recognised condition. This case, however, illustrates that precarious retropharyngeal haematoma can occur after low-energy trauma even without anticoagulation therapy. A high index of suspicion for this airway collapse is advisable in older patients.


Novel speed sintered zirconia by microwave technology.
Continuous efforts have been made to hasten the zirconia densification process without compromising properties. This study evaluated the long-term structural durability of microwave speed-sintered zirconia (MWZ) relative to a conventionally sintered zirconia (CZ). As-machined dental 3Y-TZP discs (Ø12 × 1.2 mm) were speed sintered at 1450 °C for 15 min using an industrial microwave oven, while conventional sintering was conducted in a standard dental furnace at 1530 °C for 2 h. Both were followed by natural cooling. The total sintering time was 105 min for MWZ and 600 min for CZ. Groups were compared regarding density, grain size, phase composition, and fracture resistance. Structural durability was investigated employing two fatigue protocols, step-stress and dynamic fatigue. Compared to CZ, MWZ exhibited a slightly lower density (MWZ = 5.98 g/cm3, CZ = 6.03 g/cm3), but significantly smaller grain sizes (MWZ = 0.53 ± 0.09 μm, CZ = 0.89 ± 0.10 μm), lower cubic-zirconia contents (MWZ = 15.3%, CZ = 22.7%), and poorer translucency properties (TP) (MWZ = 13 ± 1, CZ = 29 ± 0.8). However, the two materials showed similar flexural strength (MWZ = 978 ± 112 MPa, CZ = 1044 ± 161 MPa). Additionally, step-stress testing failed to capture the fatigue effect in 3Y-TZP, whereas dynamic fatigue revealed structural degradation due to moisture-assisted slow-crack-growth (SCG). Finally, MWZ possessed a slightly higher Weibull modulus (MWZ = 7.9, CZ = 6.7) but similar resistance to SCG (MWZ = 27.5, CZ = 24.1) relative to CZ. Dental 3Y-TZP with similar structural durability can be fabricated six-times faster by microwave than conventional sintering.


Mash1 and neurogenin 2 enhance survival and differentiation of neural precursor cells after transplantation to rat brains via distinct modes of action.
Neural precursor cells (NPCs) are regarded as a promising source of donor cells in transplantation-based therapies for neurodegenerative disorders. However, poor survival and limited neuronal differentiation of the transplanted NPCs remain critical limitations for developing therapeutic strategies. In this study, we investigated the effects of the proneural basic helix-loop-helix (bHLH) transcription factors Mash1 and Neurogenin 2 (Ngn2) in neuronal differentiation and survival of NPCs. Induction of Mash1 or Ngn2 expression strikingly enhanced neuronal differentiation of cultured NPCs in vitro. Ngn2-transduced NPCs underwent a rapid cell cycle arrest, which often accompanies differentiation. In contrast, cells continuously expanded upon Mash1 expression during NPC differentiation. Notably, sonic hedgehog (SHH) was upregulated by Mash1 and mediated the proliferative and survival effects of Mash1 on NPCs. Upon transplantation into adult rat brains, Mash1-expressing NPCs yielded large grafts enriched with neurons compared to control LacZ-transduced NPCs. Interestingly, enhancements in neuronal yield, as well as in donor cell survival, were also achieved by transplanting Ngn2-transduced NPCs. We show that a differentiation stage- and cell density-dependent survival effect of Ngn2 involves neurotrophin3 (NT3)/TrkC-mediated signaling. Together, these findings suggest potential benefits of bHLH gene manipulation to develop successful transplantation strategies for brain disorders.


Restaging in gynaecological cancers.
Regardless of recent technical developments in the scientific arena, stage is still the most important prognostic factor in gynaecological cancers. Surgical staging is performed in all types of gynaecologic cancers except for cervical cancer. Adjuvant therapies that contribute to survival are planned in the light of information obtained from staging procedures. Therefore, necessary information for further therapeutic management should be revealed by the end of surgical staging. A staging surgery that is not completed for any reason will not only deprive the patient of necessary treatments, but can also cause administration of unnecessary adjuvant treatments. This is especially important, given the undesired effects and cost of both chemotherapy and radiotherapy. A particularly relevant case in point is tumours that look like early stage; this is because upstaging up to 30% has been reported in ovarian and endometrial cancers. As for vulvar cancer, clinical staging has been reported to lead to about 15% over-diagnosis in comparison to surgical staging. Thus, the first step in all gynaecological cancers, except cervical cancer, should be to perform surgical staging when possible and unveil all surgical-pathological prognostic factors in the light of data obtained. Accordingly, restaging surgery should be considered in all cases that had incomplete staging. However, care should be taken to evaluate the benefits to be reaped together with the operative morbidity risk associated with the restaging procedure. This will both ensure accurate planning of postoperative treatment and provide a universal standard of approaching cancer patients and their treatments.


Cloning and characterization of the human homolog of mouse Jak2.
Members of the Jak family play a critical role in signal transduction mediated by cytokine and hormone receptors. In this study, we report the cloning and characterization of human Jak2. The predicted amino acid sequence shows 91% homology to the described murine Jak2, but with a significant difference in the extreme C-terminal sequence. Using the human cDNA as a probe, we localized the gene for human Jak2 to chromosome 9p23-24. Human Jak2 mRNA is highly expressed in the spleen, lymph nodes, and peripheral blood lymphocytes (PBLs). A polyclonal antibody raised against the unique C-terminus of human Jak2 was used to characterize Jak2 protein. Levels of Jak2 protein expression increased significantly in mitogen- and anti-IgM-stimulated B cells and to a lesser degree in activated T cells. In addition, high levels of Jak2 protein were detected in pre-B leukemia cells.


Studies on functional domains of the regulatory subunit of bovine heart adenosine 3':5'-monophosphate-dependent protein kinase.
The functional domains of the regulatory subunit of isozyme II of cAMP-dependent protein kinase were studied. It was shown using Edman degradation that the regulatory subunit contained a phosphorylated residue which was very close in primary sequence to the site most sensitive to hydrolysis by low trypsin concentrations as postulated previously (Corbin, J.D., Sugden, P.H., West, L., Flockhart, D.A., Lincoln, T.M., and McCarthy, D. (1978) J. Biol. Chem. 253, 3997-4003). Catalytic subunit incorporated 0.9 mol of 32P from [gamma-32P]ATP into a preparation of regulatory subunit that contained 1.1 mol of endogenous phosphate. After phosphorylation by the catalytic subunit, the regulatory subunit contained 2.2 mol of chemical phosphate. The effects of heat denaturation upon the rate and extent of phosphorylation of the regulatory subunit were compared with the effects of these treatments upon the cAMP binding and inhibitory domains. These data suggested that the regulatory subunit required factors in addition to an intact phosphorylatable primary sequence in order for inhibitory activity to be expressed. Such factors might be part of the secondary or tertiary structure of the protein. These studies are discussed with respect to the mechanism of inhibition of catalytic activity, and a model of the regulatory subunit structure is proposed.


Evaluation of a quality improvement intervention for labour and birth care in Brazilian private hospitals: a protocol.
In Brazilian private hospitals, caesarean section (CS) is almost universal (88%) and is integrated into the model of birth care. A quality improvement intervention, "Adequate Birth" (PPA), based on four driving components (governance, participation of women and families, reorganisation of care, and monitoring), has been implemented to help 23 hospitals reduce their CS rate. This is a protocol designed to evaluate the implementation of PPA and its effectiveness at reducing CS as a primary outcome of birth care. Case study of PPA intervention conducted in 2017/2018. We integrated quantitative and qualitative methods into data collection and analysis. For the quantitative stage, we selected a convenient sample of twelve hospitals. In each of these hospitals, we included 400 women. This resulted in a total sample of 4800 women. We used this sample to detect a 2.5% reduction in CS rate. We interviewed managers and puerperal women, and extracted data from hospital records. In the qualitative stage, we evaluated a subsample of eight hospitals by means of systematic observation and semi-structured interviews with managers, health professionals and women. We used specific forms for each of the four PPA driving components. Forms for managers and professionals addressed the decision-making process, implemented strategies, participatory process in strategy design, and healthcare practice. Forms for women and neonatal care addressed socio-economic, demographic and health condition; prenatal and birth care; tour of the hospital before delivery; labour expectation vs. real experience; and satisfaction with care received. We will estimate the degree of implementation of PPA strategies related to two of the four driving components: "participation of women and families" and "reorganisation of care". We will then assess its effect on CS rate and secondary outcomes for each of the twelve selected hospitals, and for the total sample. To allow for clinical, socio-demographic and obstetric characteristics in women, we will conduct multivariate analysis. Additionally, we will evaluate the influence of internal context variables (the PPA driving components "governance" and "monitoring") on the degree of implementation of the components "participation of women and families" and "reorganisation of care", by means of thematic content analysis. This analysis will include both quantitative and qualitative data. The effectiveness of quality improvement interventions that reduce CS rates requires examination. This study will identify strategies that could promote healthier births.


Extracting Optical Fiber Background from Surface-Enhanced Raman Spectroscopy Spectra Based on Bi-Objective Optimization Modeling.
We propose a bi-objective optimization model for extracting optical fiber background from the measured surface-enhanced Raman spectroscopy (SERS) spectrum of the target sample in the application of fiber optic SERS. The model is built using curve fitting to resolve the SERS spectrum into several individual bands, and simultaneously matching some resolved bands with the measured background spectrum. The Pearson correlation coefficient is selected as the similarity index and its maximum value is pursued during the spectral matching process. An algorithm is proposed, programmed, and demonstrated successfully in extracting optical fiber background or fluorescence background from the measured SERS spectra of rhodamine 6G (R6G) and crystal violet (CV). The proposed model not only can be applied to remove optical fiber background or fluorescence background for SERS spectra, but also can be transferred to conventional Raman spectra recorded using fiber optic instrumentation.


Entertainment-education and HIV/AIDS prevention: a field experiment in Tanzania.
Entertainment-education is the process of designing and implementing an entertainment program to increase audience members' knowledge about a social issue, create more favorable attitudes, and change their overt behaviors regarding the social issue. The results of a field experiment in Tanzania to measure the effects of a long-running entertainment-education radio soap opera, Twende na Wakati (Let's Go with the Times), on knowledge, attitudes, and adoption of human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) prevention behaviors are presented. Multiple independent measures of effects and the experimental design of this study confer strong internal and external validity regarding the results of this investigation. The effects of the radio program in Tanzania include (1) a reduction in the number of sexual partners by both men and women, and (2) increased condom adoption. The radio soap opera influenced these behavioral variables through certain intervening variables, including (1) self-perception of risk of contracting HIV/AIDS, (2) self-efficacy with respect to preventing HIV/AIDS, (3) interpersonal communication about HIV/AIDS, and (4) identification with, and role modeling of, the primary characters in the radio soap opera.


Surface Structure and Stability of Partially Hydroxylated Silica Surfaces.
Surface energies of silicates influence crack propagation during brittle fracture and decrease with surface relaxation caused by annealing and hydroxylation. Molecular-level simulations are particularly suited for the investigation of surface processes. In this work, classical MD simulations of silica surfaces are performed with two force fields (ClayFF and ReaxFF) to investigate the effect of force field reactivity on surface structure and energy as a function of surface hydroxylation. An unhydroxylated fracture surface energy of 5.1 J/m2 is calculated with the ClayFF force field, and 2.0 J/m2 is calculated for the ReaxFF force field. The ClayFF surface energies are consistent with the experimental results from double cantilever beam fracture tests (4.5 J/m2), whereas ReaxFF underestimated these surface energies. Surface relaxation via annealing and hydroxylation was performed by creating a low-energy equilibrium surface. Annealing condensed neighboring siloxane bonds increased the surface connectivity, and decreased the surface energies by 0.2 J/m2 for ClayFF and 0.8 J/m2 for ReaxFF. Posthydroxylation surface energies decreased further to 4.6 J/m2 with the ClayFF force field and to 0.2 J/m2 with the ReaxFF force field. Experimental equilibrium surface energies are ∼0.35 J/m2, consistent with the ReaxFF force field. Although neither force field was capable of replicating both the fracture and equilibrium surface energies reported from experiment, each was consistent with one of these conditions. Therefore, future computational investigations that rely on accurate surface energy values should consider the surface state of the system and select the appropriate force field.


Susceptibility of Gram-positive bacteria from ICU patients in UK hospitals to antimicrobial agents.
Microbiologists in 25 sentinel laboratories were each asked to refer up to 100 clinically-significant Gram-positive bacteria isolated from consecutive intensive care unit (ICU) patients. A total of 1595 isolates were collected from patients in 23 hospitals; these included Staphylococcus aureus (47.6%), coagulase-negative staphylococci (CNS) (30.6%), enterococci (14.3%), pneumococci (2.8%) and other streptococci (3.5%). A few coryneforms, other bacilli and a Nocardia sp. were also collected. Rates of oxacillin resistance among S. aureus and CNS isolates were 59.3 and 78.5%, respectively. Vancomycin-resistant S. aureus were not detected, although two isolates (0.3%) were resistant to teicoplanin [minimum inhibitory concentrations (MICs) 8 mg/L]. In contrast, 13.7% of CNS were teicoplanin resistant (MICs 8-32 mg/L) and 1.2% were resistant to vancomycin. Among the enterococci, 72.5% were Enterococcus faecalis and 24.5% were Enterococcus faecium, the remainder including isolates of Enterococcus casseliflavus or Enterococcus gallinarum. Eighteen percent of E. faecium isolates were vancomycin-resistant, compared with only 3% of E. faecalis isolates. Rates of high-level gentamicin resistance in E. faecalis and E. faecium were 40 and 25%, respectively. Nine percent of pneumococci and streptococci were resistant to penicillin, with 7 and 11%, respectively, resistant to erythromycin. None of the isolates showed resistance to linezolid, with the MICs for the entire study population falling in the range of 0.5-4 mg/L.


Detection of Candidatus Mycoplasma haemobos in cattle with anaemia.
Haemotrophic mycoplasmas are unculturable eperythrocytic pathogens that are found in a wide range of domestic and wild animals. In this study an outbreak of haemotrophic mycoplasmosis in cattle herds in Northern Germany is reported. Affected animals exhibited anaemia and depression and infection was confirmed following microscopic examination of blood smears and on PCR. Sequence analysis indicated that in addition to infection with Mycoplasma wenyonii, animals were infected with a novel bovine haemotrophic mycoplasma Candidatus Mycoplasma haemobos.


Differential effects of glycogen synthase kinase 3 (GSK3) inhibition by lithium or selective inhibitors in the central nervous system.
Glycogen synthase kinase (GSK3) is a constitutively active serine-threonine kinase associated to neurological and psychiatric disorders. GSK3 inhibition is considered a mediator of the efficacy of the mood-stabiliser lithium. This study aimed at comparing the central nervous system effect of lithium with the selective GSK3 inhibitors AZ1080 and compound A in biochemical, cellular, and behavioural tests. Collapsin response mediator protein 2 is a neuron-specific GSK3 substrate. Lithium, AZ1080, and compound A inhibited its phosphorylation in rat primary neurons with different pIC50. After systemic treatments with lithium or GSK3 inhibitors to assess specific functional responses, phosphorylation was unchanged in adult rat brain, while it was strongly inhibited by GSK3 inhibitors in pups, differently from lithium. Lithium may exert neurotrophic effect by increasing brain-derived neurotrophic factor (BDNF) levels: in the present experimental conditions, lithium exerted opposite effects on plasma BDNF levels compared to GSK3 inhibitors, suggesting this effect might not be necessarily mediated by GSK3 inhibition alone. While plasma thyroid-stimulating hormone and luteinising hormone were not affected by lithium, they were decreased by selective inhibitors. GH and prolactin displayed similar responses towards reduction. Follicle-stimulating hormone levels were not altered by treatments, whereas melatonin was specifically increased by AZ1080. Lithium impaired mouse spontaneous locomotion and decreased amphetamine-induced hyper-locomotion. AZ1080 had no effects on locomotion, while compound A reduced spontaneous locomotor activity without effects on amphetamine-induced hyper-locomotion. The present results indicate that a broad correlation between the effects of lithium and selective GSK3 inhibitors could not be devised, suggesting alternative mechanisms, whereas overlapping results could be obtained in specific assays.


Identification of apolipoprotein A-I in BALF as a biomarker of sarcoidosis.
Background: Sarcoidosis goes into remission in two-thirds of patients with sarcoidosis, but about 20 % of patients develop pulmonary fibrosis. The mechanisms of pulmonary fibrosis in sarcoidosis and differences in pathogenesis between clinical stages are still unclear. Objectives: The aim of this study was investigating proteins associated with clinical stages by comparing bronchoalveolar lavage fluid (BALF) protein between stage I and stage IV using proteome analysis. Methods: Proteomic differences in BALF were compared between stage I and stage IV by examining BALF from 8 stage I patients and 5 stage IV patients by two-dimensional gel electrophoresis and mass spectrometry. Results: In individual comparisons of BALF samples, the levels of apolipoprotein (Apo) A-I fragment, fibrinogen γ chain, calcyphosine, complement C3, and surfactant protein A were significantly higher in stage I than in stage IV. In contrast, none of the proteins examined significantly higher in stage IV than in stage I. To confirm the results of Apo A-I in the BALF proteome, we performed enzyme-linked immunosorbent assay (ELISA) in a larger group. The concentration of BALF Apo A-I was significantly higher in stage I patients than in stage IV patients (0.70 [0.13-0.89] vs. 0.15 [0.08-0.21] ng/μg protein, p=0.003). Conclusion: The involvement of BALF Apo A-I in sarcoidosis may differ between stage I and stage IV. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 5-15).


Identification and characterization of ace1-type acetylcholinesterase in insecticide-resistant and -susceptible Propylaea japonica (Thunberg).
Characterization and gene cloning of acetylecholinesterase (AChE) in the insecticide-resistant (R) and -susceptible (S) insects have been reported in the past. However, the studies focused mostly on herbivorous pests, rather than predacious species, such as ladybird beetles. Using R and S Propylaea japonica (thunberg), a full-length cDNA sequence (2928 bp) of the ace1-type AChE gene was determined for the first time. The ace1 encoding a protein of 645 amino acids contained typical conserved motifs, such as FGESAG domains, catalytic triad, acyl pocket, oxyanino hole, choline binding site, peripheral anionic site, omega loop and conserved aromatic residues. R P. japonica displayed 50-times greater resistance to chlorpyrifos or mathamidophos with a significantly lower AChE sensitivity to paraoxon, malaoxon, chlorpyrifos or methamidophos than its S counterpart. Five amino acids in the ace1 of R P. japonica differed from those found in S P. japonica. One of them, F358S, located in the acyl-binding pocket, might play a crucial role in the resistance of the insect to organophosphates (OPs). Whereas, K493E and I538V, which were close to some of the conserved aromatic amino acids (i.e., H509, Y511, and W499) in the gorge, and G571R and T576A near C593 that formed the disulfide bonds with C471, might also involve in the change of insecticide resistance in P. japonica. AChE insensitivity and amino acid replacements, particularly F358S, might be the determining factors in the alteration of OPs-resistance in P. japonica.


Characteristics associated with anxiety, depressive symptoms, and quality-of-life in a large cohort of implantable cardioverter defibrillator recipients.
Although most patients with implantable cardioverter defibrillators (ICDs) adjust well, some have considerable psychological distress. Factors associated with psychological adjustment in ICD-recipients are still not well understood. Our purpose was to describe quality-of-life (QoL) and prevalence of self-reported symptoms of anxiety and depression in a large national cohort of ICD-recipients, and to determine socio-demographic, clinical, and ICD-related factors associated with these variables. A cross-sectional, correlational design was used. All eligible adult ICD-recipients in the Swedish ICD- and Pacemaker Registry were invited to participate. Symptoms of anxiety and depression were measured using the Hospital Anxiety and Depression Scale (HADS), and QoL with the EuroQol-5D. A total of 3067 ICD-recipients (66±11years, 80% male) were included. The mean HADS score was 3.84±3.70 for anxiety symptoms and 2.99±3.01 for symptoms of depression. The mean EQ-5D index score was 0.82±0.21. The probability of symptoms of anxiety and depression was associated with younger age, living alone, and a previous history of myocardial infarction or heart failure. Additionally, female ICD-recipients had a higher probability of symptoms of anxiety. A higher level of ICD-related concerns was most prominently related to symptoms of anxiety, depressive symptoms and poorer QoL, while number of shocks, ICD-indication and time since implantation were not independently related. In this large cohort of ICD-recipients, the association of ICD-related concerns with symptoms of anxiety, depressive symptoms, and poor QoL suggests that ICD specific factors should be addressed in order to improve outcomes.


Comparative functional anatomy of hindlimb muscles and bones with reference to aquatic adaptation of the sea otter.
Although the sea otter (Enhydra lutris) is a complete aquatic species, spending its entire life in the ocean, it has been considered morphologically to be a semi-aquatic animal. This study aimed to clarify the unique hindlimb morphology and functional adaptations of E. lutris in comparison to other Mustelidae species. We compared muscle mass and bone measurements of five Mustelidae species: the sea otter, Eurasian river otter (Lutra lutra), American mink (Neovison vison), Japanese weasel (Mustela itatsi) and Siberian weasel (M. sibirica). In comparison with the other 4 species, E. lutris possessed significantly larger gluteus, popliteus and peroneus muscles, but smaller adductor and ischiopubic muscles. The popliteus muscle may act as a medial rotator of the crus, and the peroneus muscle may act as an abductor of the fifth toe and/or the pronator of the foot. The bundles of the gluteus superficialis muscle of E. lutris were fused with those of the tensor fasciae latae muscle and gluteofemoralis muscles, and they may play a role in femur abduction. These results suggest that E. lutris uses the abducted femur, medially rotated crus, eversion of the ankle and abducted fifth digit or extended interdigital web as a powerful propulsion generator. Therefore, we conclude that E. lutris is a complete aquatic animal, possessing differences in the proportions of the hindlimb muscles compared with those in other semi-aquatic and terrestrial mustelids.


Clinical, histologic, microbiologic, and biochemical characterization of the causative agent of bacillary (epithelioid) angiomatosis: a rickettsial illness with features of bartonellosis.
It has been suggested that bacillary (epithelioid) angiomatosis (BEA) is a manifestation of cat scratch disease (CSD). Because of clinical similarity between this condition and the verruga peruana phase of bartonellosis, we sought to further characterize this disease as well as its causative agent and to compare it to bartonellosis. We isolated a small flagellated pleomorphic bacillus from skin lesions of two patients with BEA. Organisms were stained successfully with Warthin-Starry silver stains, but immunohistochemistry failed to demonstrate binding with a polyclonal antibody directed against the cat scratch bacillus. Whole cell fatty-acid gas chromatography performed on both BEA organisms and Bartonella bacilliformis demonstrated marked similarity between the two. Electron microscopy of BEA organisms in tissue and in suspension revealed features characteristic of a gram negative bacillus. Based on these findings, we propose that this unusual rickettsial infectious disease with vascular proliferation may represent an unusual variant of infection with a bartonella-like organism rather than a manifestation of cat scratch disease.


MIB-MIP is a mycoplasma system that captures and cleaves immunoglobulin G.
Mycoplasmas are "minimal" bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB-IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB-MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas.


Delineation of T-wave in ECG by wavelet transform using multiscale differential operator.
This paper proposes an improved wavelet-based method for the delineation of T wave in the electrocardiogram by using multiscale dif-operator (MDO). MDO was employed to automatically categorize the T wave into one of the three categories of T wave morphologies and to improve the precision of T wave delineation for each category. The new algorithm was evaluated on QT database (QTDB) and new annotations of 2160 beats from QTDB were performed by two cardiologists. To evaluate the performance of the wave delineation, the time differences between automatic detection versus cardiologists' annotations and intercardiologist differences were measured. The new algorithm can attain better results than the previous methods by achieving the smallest standard deviation of the differences and qualifying the strict error criterion for T-off measurement. This new algorithm also exhibited excellent T wave categorization agreement with the cardiologists, resulting in kappa values above 0.75.


Selective optogenetic stimulation of the retrotrapezoid nucleus in sleeping rats activates breathing without changing blood pressure or causing arousal or sighs.
Combined optogenetic activation of the retrotrapezoid nucleus (RTN; a CO2/proton-activated brainstem nucleus) with nearby catecholaminergic neurons (C1 and A5), or selective C1 neuron stimulation, increases blood pressure (BP) and breathing, causes arousal from non-rapid eye movement (non-REM) sleep, and triggers sighs. Here we wished to determine which of these physiological responses are elicited when RTN neurons are selectively activated. The left rostral RTN and nearby A5 neurons were transduced with channelrhodopsin-2 (ChR2(+)) using a lentiviral vector. Very few C1 cells were transduced. BP, breathing, EEG, and neck EMG were monitored. During non-REM sleep, photostimulation of ChR2(+) neurons (20s, 2-20 Hz) instantly increased V̇e without changing BP (13 rats). V̇e and BP were unaffected by light in nine control (ChR2(-)) rats. Photostimulation produced no sighs and caused arousal (EEG desynchronization) more frequently in ChR2(+) than ChR2(-) rats (62 ± 5% of trials vs. 25 ± 2%; P < 0.0001). Six ChR2(+) rats then received spinal injections of a saporin-based toxin that spared RTN neurons but destroyed surrounding catecholaminergic neurons. Photostimulation of the ChR2(+) neurons produced the same ventilatory stimulation before and after lesion, but arousal was no longer elicited. Overall (all ChR2(+) rats combined), ΔV̇e correlated with the number of ChR2(+) RTN neurons whereas arousal probability correlated with the number of ChR2(+) catecholaminergic neurons. In conclusion, RTN neurons activate breathing powerfully and, unlike the C1 cells, have minimal effects on BP and have a weak arousal capability at best. A5 neuron stimulation produces little effect on breathing and BP but does appear to facilitate arousal.


Stigma against mental health disorders in Nepal conceptualised with a 'what matters most' framework: a scoping review.
Stigma related to mental disorders is a barrier to quality mental healthcare. This scoping review aimed to synthesise literature on stigma related to mental disorders in Nepal to understand stigma processes. The anthropological concept of 'what matters most' to understand culture and stigma was used to frame the literature on explanatory models, manifestations, consequences, structural facilitators and mitigators, and interventions. We conducted a scoping review with screening guided by the Preferred Reporting Items for Systematic Review and Meta-analysis Extension for Scoping Reviews (PRISMA-ScR). A structured search was done using three international databases (PsycINFO, Medline and Web of Science), one Nepali database (NepJol) and cross-referencing for publications from 1 January 2000 through 24 June 2020. The search was repeated to include structural stigma-related terms. Quality of quantitative studies was assessed using the Systematic Assessment of Quality in Observational Research (SAQOR) tool. The review was registered through the Open Science Framework (OSF) (osf.io/u8jhn). The searches yielded 57 studies over a 20-year period: 19 quantitative, 19 qualitative, nine mixed methods, five review articles, two ethnographies and three other types of studies. The review identified nine stigma measures used in Nepal, one stigma intervention, and no studies focused on adolescent and child mental health stigma. The findings suggest that 'what matters most' in Nepali culture for service users, caregivers, community members and health workers include prestige, productivity, privacy, acceptance, marriage and resources. Cultural values related to 'what matters most' are reflected in structural barriers and facilitators including lack of policies, programme planning and resources. Most studies using quantitative tools to assess stigma did not describe cultural adaptation or validation processes, and 15 out of the 18 quantitative studies were 'low-quality' on the SAQOR quality rating. The review revealed clear gaps in implementation and evaluation of stigma interventions in Nepal with only one intervention reported, and most stigma measures not culturally adapted for use. As stigma processes are complex and interlinked in their influence on 'what matters most' and structural barriers and facilitators, more studies are required to understand this complexity and establish effective interventions targeting multiple domains. We suggest that stigma researchers should clarify conceptual models to inform study design and interpretations. There is a need to develop procedures for the systematic cultural adaptation of stigma assessment tools. Research should be conducted to understand the forms and drivers of structural stigma and to expand intervention research to evaluate strategies for stigma reduction.


In vivo evaluation of the "TinyPump" as a pediatric left ventricular assist device.
Pediatric patients with end-stage heart failure require mechanical circulatory support (MCS) just as adults do. In order to meet the special requirements for neonates' and infants' MCS, pediatric circulatory support devices must be compact with low priming volume, easily controllable with low flow, less traumatic for blood cells and tissues, and biocompatible with minimum anticoagulation. We have designed and developed a miniature rotary centrifugal blood pump, "TinyPump," with a priming volume of 5 mL, which has already demonstrated its controllable performance for low flow and durability in vitro. To evaluate the feasibility of the TinyPump as a left ventricular assist device (LVAD) suitable for neonates and infants, we have examined the biocompatibility and hemodynamic performance of the TinyPump in a pediatric animal model using Shiba goats. The TinyPump is a miniaturized centrifugal pump weighing 150 g comprising a disposable pump head with a 30-mm diameter impeller having six straight-vanes and a reusable motor driver. The impeller in the pump head is supported by a hydrodynamic bearing at its center and is driven by radial magnetic force coupled to the motor driver. TinyPump implantations were performed in 22 Shiba goats (17 female and 5 male), with body weights ranging from 8.4 to 27.2 kg. Under gas anesthesia, via left lateral thoracotomy, a 22 Fr inflow cannula was inserted through the left ventricular apex, while a 6-mm outflow graft was anastomosed to the descending aorta, which were then connected to a TinyPump mounted on the animal's back. Postoperative hemodynamic monitoring included heart rate, arterial and central venous pressure, pump flow, and rotation speed. Target pump flow in all animals was maintained at 0.9 ± 0.1 L/min, which is approximately half the normal pulmonary artery flow measured in control animals. Blood samples were collected to evaluate peripheral organ functions, hemolysis, and thrombosis. Goats were divided into three groups-acute phase (6 h; n = 4), subchronic phase (6 h 2 postoperative days [POD]; n = 11), and chronic phase (3 POD-16 POD; n = 8)-based on their survival duration. In the early experiments, hemolysis and thrombi formation at the impeller bearing resulted in termination of the study. Subsequent modifications of the bearing design, pump housing design, and magnetic coupling force helped to minimize the hemolysis and thrombi formation, prolonging the survival duration of the Shiba goats to 2 weeks with minimum adverse effects on the blood components and organ functions. With further experiments and improvements in pump durability and hemocompatibility, the TinyPump can serve as a suitable circulatory support device for neonates and infants bridging to heart transplantation as well as to heart recovery.


Assessment of the potential of insecticides, emulsifiers, and solvent mixtures to enhance viral infection in cultured mammalian cells.
Various insecticides, solvents, emulsifiers, and mixtures thereof were tested to determine whether these compounds are capable of enhancing the sensitivity of cultured mammalian cells to infection with vesicular stomatitis virus. None of 42 compounds tested significantly enhanced the viral infection.


Transvaginal Ultrasound Can Accurately Predict the American Society of Reproductive Medicine Stage of Endometriosis Assigned at Laparoscopy.
To evaluate the diagnostic accuracy of transvaginal ultrasound in predicting a laparoscopic, surgically assigned, revised American Society of Reproductive Medicine (ASRM) endometriosis stage. A multicenter, retrospective, diagnostic accuracy study. The patients visited 1 of 2 academic gynecologic ultrasound units and underwent laparoscopy led by 1 of 6 surgeons in metropolitan Sydney, Australia, between 2016 and 2018. Patients with suspected endometriosis (n = 204). Ultrasound followed by laparoscopy. Surgical cases were identified. The preoperative ultrasound report and surgical operative notes were each used to retrospectively assign an ASRM score and stage. The breakdown of surgical findings was as follows: ASRM 0 (i.e., no endometriosis), 24/204 (11.8%); ASRM 1, 110/204 (53.9%); ASRM 2, 22/204 (10.8%); ASRM 3, 16/204 (7.8%); ASRM 4, 32 204 (15.7%). The overall accuracy of ultrasound in predicting the surgical ASRM stage was as follows: ASRM 1, 53.4%; ASRM 2, 93.8%; ASRM 3, 89.7%; ASRM 4, 93.1%; grouped ASRM 0, 1, and 2, 94.6%; and grouped ASRM 3 and 4 of 94.6%. Ultrasound had better test performance in higher disease stages. When the ASRM stages were dichotomized, ultrasound had sensitivity and specificity of 94.9% and 93.8%, respectively, for ASRM 0, 1, and 2 and of 93.8% and 94.9%, respectively, for ASRM 3 and 4. Ultrasound has high accuracy in predicting the mild, moderate, and severe ASRM stages of endometriosis and can accurately differentiate between stages when ASRM stages are dichotomized (nil/minimal/mild vs moderate/severe). This can have major positive implications on patient triaging at centers of excellence in minimally invasive gynecology for advanced-stage endometriosis.


Chronic electrical stimulation increases MCT1 and lactate uptake in red and white skeletal muscle.
We examined whether chronic stimulation of red and white rat muscles increased the concentrations of the monocarboxylate transporter MCT1. Red and white tibialis anterior (RTA and WTA, respectively) and extensor digitorum longus (EDL) muscles were chronically stimulated via the peroneal nerve for 7 days. Stimulated and contralateral control muscles were examined for MCT1 content, L-lactate uptake, lactate dehydrogenase (LDH) isoforms, and muscle fiber composition. MCT1 was 1.5 times greater in stimulated RTA, 3 times greater in stimulated WTA, and 1.9 times greater in stimulated EDL compared with respective control muscles (P < 0.05). L-Lactate uptake increased in all stimulated muscles (P < 0.05), and this was highly correlated with the increase in MCT1 (r = 0.96). The heart-type LDH (H-LDH) subunits also increased in all stimulated muscles (P < 0.05). The H-LDH subunits correlated highly with MCT1 in the muscles (r = 0.83). There was no change in muscle-type LDH subunits (P > 0.05). There were negligible alterations in muscle fiber composition in the stimulated muscles, suggesting that the increase in MCT1 was independent of changes in muscle fiber composition. These studies are the first to demonstrate that chronic muscle contraction increases MCT1 concentrations in both red and white skeletal muscles.


Comparison of biometric measurements obtained by the Verion Image-Guided System versus the auto-refracto-keratometer.
To compare the biometric measurements obtained from the Verion Image-Guided System to those obtained by auto-refracto-keratometer in normal eyes. This is a prospective, observational, comparative study conducted at the Asociación para Evitar la Ceguera en México I.A.P., Mexico. Three sets of keratometry measurements were obtained using the image-guided system to assess the coefficient of variation, the within-subject standard deviation and intraclass correlation coefficient (ICC). A paired Student t test was used to assess statistical significance between the Verion and the auto-refracto-keratometer. A Pearson's correlation coefficient (r) was obtained for all measurements, and the level of agreement was verified using Bland-Altman plots. The right eyes of 73 patients were evaluated by each platform. The Verion coefficient of variation was 0.3% for the flat and steep keratometry, with the ICC being greater than 0.9 for all parameters measured. Paired t test showed statistically significant differences between groups (P = 0.0001). A good correlation was evidenced for keratometry values between platforms (r = 0.903, P = 0.0001 for K1, and r = 0.890, P = 0.0001). Bland-Altman plots showed a wide data spread for all variables. The image-guided system provided highly repeatable corneal power and keratometry measurements. However, significant differences were evidenced between the two platforms, and although values were highly correlated, they showed a wide data spread for all analysed variables; therefore, their interchangeable use for biometry assessment is not advisable.


Effect of bolus volume and flow time on temporospatial coordination in oropharyngeal pressure production in healthy subjects.
The effects of bolus volume and flow time on the sequential coordination of tongue pressure (TP) and pharyngeal pressure (PP), which are important in the biomechanics of swallowing, are unclear. In this study, we measured TP and PP simultaneously in 10 healthy adults at multiple points during dry swallowing and the swallowing of 5 ml and 15 ml of liquids with different viscosities, and investigated changes in the timing of the onset, peak, and offset of these pressures. TP was measured using a sensor sheet system with five measuring points on the hard palate, and PP was measured using a manometry catheter with four measuring points. The order and correlations of sequential events, such as onset, peak, and offset times of pressure production, at each pressure measuring point were analyzed on the synchronized waveforms. We found that the differences between the TP and PP onset times decreased when the bolus volume was larger. The change in bolus volume had very little effect on peak time or offset time. The flow time of the bolus affected the appearance of onset and peak time for both TP and PP. A time difference between TP and PP emerged as the flow time increased, with TP starting to appear before PP. This may be the first detailed analysis of pressure-flow dynamics that treats the mouth and pharynx as a single functional unit. We believe that our analysis is an important step toward extending future research to include a wider range of age groups and dysphagia patients.


Immunosuppression and Multiple Primary Malignancies in Kidney-Transplanted Patients: A Single-Institute Study.
Immunodeficiency is associated with higher cancer incidence. However, it is unknown whether there is a link between immunodeficiency and development of multiple primary malignancies. In the present study we analyse this link focusing on kidney-transplanted patients, as they are at higher risk of developing cancer due to the chronic assumption of immunosuppressants. We followed up 1200 patients who underwent kidney transplantation between 1980 and 2012. A total of 77/1200 kidney-transplanted patients developed cancer and 24 of them developed multiple cancers. Most multiple cancers were synchronous with a nonsignificant association between cancer and rejection episodes. In the general cancer population, one-ninth of patients are at higher risk of developing a second tumor over a lifetime; hence it would be reasonable to conclude that, from a merely theoretical and statistical viewpoint, long-term transplanted patients potentially have a higher risk of developing MPMs. However, data did not confirm this assumption, probably because these patients die before a second primary malignancy appears. Despite many observations on the increased incidence of different tumor types in immunodeficient patients and despite immunosuppression certainly being a predisposing factor for the multicancer syndrome, data so far are not robust enough to justify a correlation between immunodeficiency and multiple primary malignancies in transplanted patients.


[Larvae of Anisakidae nematodes and Trypanorhyncha cestodes of public health importance in Aluterus monoceros (Linnaeus, 1758) in Rio de Janeiro State, Brazil].
One hundred specimens of unicorn leatherjacket, Aluterus monoceros purchased from markets of municipalities of Niterói and Rio de Janeiro from May to August 2006. The fishes were measured, necropsied, fileted and analysed their organs. Sixteen fishes were parasitized by nematode Anisakidae: Anisakis spp. and Contracaecum sp. with respectively, 1 and 16% of prevalence, 2 and 3.31 of mean intensity, and 0.02 and 0.53 of mean abundance. Two larvae of Anisakis sp. were found in mesentery of one fish and Contracaecum sp. was found in liver and mesentery with 1 to 9 specimens of range of infection. Fifty-one fishes were parasitized on the liver and mesentery by metacestodes of Trypanorhyncha. The collected species were Floriceps saccatus and Callitetrarhynchus speciosus with respectively, 45 and 6% of prevalence, 3.17 and 2.83 of mean intensity, and 1.43 and 0.06 of mean abundance, the range of infection by F. saccatus was 1 to 20 and by C. speciosus was 1 to 5. Anisakis sp. and these two species of Trypanothyncha were reported in A. monoceros for the first time.


Localization-delocalization transition in Hessian matrices of topologically disordered systems.
Using the level-spacing (LS) statistics, we have investigated the localization-delocalization transitions (LDTs) in Hessian matrices of a simple fluid with short-ranged interactions. The model fluid is a prototype of topologically disordered systems and its Hessian matrices are recognized as an ensemble of Euclidean random matrices with elements subject to several kinds of constraints. Two LDTs in the Hessian matrices are found, with one in the positive-eigenvalue branch and the other in the negative-eigenvalue one. The locations and the critical exponents of the two LDTs are estimated by the finite-size scaling for the second moments of the nearest-neighbor LS distributions. Within numerical errors, the two estimated critical exponents are almost coincident with each other and close to that of the Anderson model (AM) in three dimensions. The nearest-neighbor LS distribution at each LDT is examined to be in a good agreement with that of the AM at the critical disorder. We conclude that the LDTs in the Hessian matrices of topologically disordered systems exhibit the critical behaviors of orthogonal universality class.


[Meiosis-like reduction in the culture system for somatic embryogenesis of Arabidopsis thaliana].
The somatic meiosis-like reduction was observed in some cells in the embryogenic calli of Arabidopsis thaliana. One type was somatic chromosome reductional grouping, in which the chromosomes in a cell either separated into two groups, or separated into more than two groups at prophase or metaphase. The chromosomes separated directly, the morphology of the chromosomes did not show any role of the spindle fibers. Chromosome reductional grouping happened more frequently in polyploid cells. The other type was somatic meiossi which was analogous to that existed in the process of gametogenesis, and characterized by the pairing and synapsis of homologues chromosomes. In the embryogenic calli cultured on dedifferentiation medium, about 3% of the observed cells contained haploid number of chromosomes, while the tetrad cells occurred at a frequency of about 4%. The somatic meiosis-like reduction could result in the recombination of chromosomes in the cells generated by these processes. The roles of somatic meiosis-like reduction in the somatic embryogenesis and somaclonal variations were discussed.


[Nursing Experience of Caring for a Patient With COVID-19 During Isolation].
The ongoing coronavirus disease-2019 (COVID-19) epidemic continues to have a global impact. This article describes the author's experience providing nursing care to a female patient who was diagnosed with COVID-19 after returning to Taiwan from overseas. During the patient's period of isolation (February 5th to February 29th, 2020), the author used Roy's adaptation model to perform first-level holistic physical, psychological, and social nursing assessments and collected health-problem information using face-to-face interviews, telephone interviews, and observations via a remote monitoring system. A second level of assessment confirmed that the problems faced by the patient included (1) existing infections related to COVID-19 and (2) anxiety related to uncertainties about disease prognosis, forgiveness from the family, and potential violations of personal privacy by the media. Due to the special nature of the isolation ward, the author used a remote physiological monitoring system to monitor the vital signs, fever, and shortness of breath status of the patient to quickly decrease her physical discomfort and to improve her self-care ability during hospitalization. Environmental cleanliness was strictly maintained to reduce the risk of cross-infection and ensure patient safety. To alleviate patient anxiety, the author established a good therapeutic interpersonal relationship with the patient by making 10-60 minutes of caring calls to her each day, by providing individual care measures, and by using the Internet to play audio and video to teach mindfulness meditation. Caring for COVID-19 cases is a completely new experience. The author hopes that this experience may be used as a reference for caregivers.


[Toxicity of spiromesifen to the developmental stages of Bactericera cockerelli (Sulc) (Hemiptera: Triozidae)].
Spiromesifen is an insecticide that inhibits the synthesis of lipids and, in Mexico, its use against the Tomato-Potato Psyllid, Bactericera cockerelli (Sulc), on chili pepper (Capsicum annum), tomato (Lycopersicon sculentum) and potato (Solanum tuberosum) began in 2005; however more information is needed to understand its toxicity on this insect pest. The aim of this research was to determine the toxicity of spiromesifen against each of the biological stages of tomato-potato psyllid, its effect on fertility and viability of eggs deposited by treated females, as well as the female preference to lay eggs on treated and non treated plants. The relative toxicity at 95% mortality (highest LC95 value /LC95 value of the respective biological stage) of spiromesifen in egg, nymph 1, nymph 2, nymph 3, nymph 4, and nymph 5 were 517.5; 31316.2; 2950.1; 315.6; 18.2 and 1-fold, respectively. There were no differences in the toxicity of spiromesifen between adult males and females. The number of laid eggs was reduced as the spiromesifen concentration used to treat female increased and egg hatch was reduced in all tested doses. In the "no choice" test, females deposited 38.6+/-2.01 eggs by leaf of non treated chili pepper type jalapeño, while in the treated with 360 mg L(-1) we observed 0.3+/-0.08 eggs by leaf. In the "choice" test, the oviposition decreased as the dose increased. There were no eggs on plants treated with 2400 mg L(-1) of spiromesifen.


Quantitative phosphoproteomics of cytotoxic T cells to reveal protein kinase d 2 regulated networks.
The focus of the present study was to characterize the phosphoproteome of cytotoxic T cells and to explore the role of the serine threonine kinase PKD2 (Protein Kinase D2) in the phosphorylation networks of this key lymphocyte population. We used Stable Isotope Labeling of Amino acids in Culture (SILAC) combined with phosphopeptide enrichment and quantitative mass-spectrometry to determine the impact of PKD2 loss on the cytotoxic T cells phosphoproteome. We identified 15,871 phosphorylations on 3505 proteins in cytotoxic T cells. 450 phosphosites on 281 proteins were down-regulated and 300 phosphosites on 196 proteins were up-regulated in PKD2 null cytotoxic T cells. These data give valuable new insights about the protein phosphorylation networks operational in effector T cells and reveal that PKD2 regulates directly and indirectly about 5% of the cytotoxic T-cell phosphoproteome. PKD2 candidate substrates identified in this study include proteins involved in two distinct biological functions: regulation of protein sorting and intracellular vesicle trafficking, and control of chromatin structure, transcription, and translation. In other cell types, PKD substrates include class II histone deacetylases such as HDAC7 and actin regulatory proteins such as Slingshot. The current data show these are not PKD substrates in primary T cells revealing that the functional role of PKD isoforms is different in different cell lineages.


Identification of N-Phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine Derivatives as Novel, Potent, and Selective NF-κB Inducing Kinase (NIK) Inhibitors for the Treatment of Psoriasis.
A series of N-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine derivatives with NF-κB inducing kinase (NIK) inhibitory activity were obtained through structure-based drug design and synthetic chemistry. Among them, 4-(3-((7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-morpholinophenyl)-2-(thiazol-2-yl)but-3-yn-2-ol (12f) was identified as a highly potent NIK inhibitor, along with satisfactory selectivity. The pharmacokinetics of 12f and its ability to inhibit interleukin 6 secretion in BEAS-2B cells were better than compound 1 developed by Amgen. Oral administration of different doses of 12f in an imiquimod-induced psoriasis mouse model showed effective alleviation of psoriasis, including invasive erythema, swelling, skin thickening, and scales. The underlying pathological mechanism involved attenuation of proinflammatory cytokine and chemokine gene expression, and the infiltration of macrophages after the treatment of 12f. This work provides a foundation for the development of NIK inhibitors, highlighting the potential of developing NIK inhibitors as a new strategy for the treatment of psoriasis.


Role of glucocorticoid-induced leucine zipper (GILZ) in bone acquisition.
Glucocorticoids (GCs) have both anabolic and catabolic effects on bone. However, no GC anabolic effect mediator has been identified to date. Here we show that targeted expression of glucocorticoid-induced leucine zipper (GILZ), a GC anti-inflammatory effect mediator, enhances bone acquisition in mice. Transgenic mice, in which the expression of GILZ is under the control of a 3.6-kb rat type I collagen promoter, exhibited a high bone mass phenotype with significantly increased bone formation rate and osteoblast numbers. The increased osteoblast activity correlates with enhanced osteogenic differentiation and decreased adipogenic differentiation of bone marrow stromal cell cultures in vitro. In line with these changes, the mRNA levels of key osteogenic regulators (Runx2 and Osx) increased, and the level of adipogenic regulator peroxisome proliferator-activated receptor (PPAR) γ2 decreased significantly. We also found that GILZ physically interacts with C/EBPs and disrupts C/EBP-mediated PPARγ gene transcription. In conclusion, our results showed that GILZ is capable of increasing bone acquisition in vivo, and this action is mediated via a mechanism involving the inhibition of PPARγ gene transcription and shifting of bone marrow MSC/progenitor cell lineage commitment in favor of the osteoblast pathway.


Nonwage losses associated with occupational injury among health care workers.
To examine nonwage losses after occupational injury among health care workers and the factors associated with the magnitude of these losses. Inception cohort of workers filing an occupational injury claim in a Canadian province. Worker self-reports were used to calculate (1) the nonwage economic losses in 2010 Canadian dollars, and (2) the number of quality-adjusted days of life lost on the basis of the EuroQOL Index. Most workers (84%; n = 123) had musculoskeletal injuries (MSIs). Each MSI resulted in nonwage economic losses of Can$3131 (95% confidence interval, Can$3035 to Can$3226), lost wages of Can$5286, and 7.9 quality-adjusted days of life lost within 12 weeks after injury. Losses varied with type of injury, region of the province, and occupation. Non-MSIs were associated with smaller losses. These estimates of nonwage losses should be considered in workers' injury compensation policies and in economic evaluation studies.


Magnesium limitation is an environmental trigger of the Pseudomonas aeruginosa biofilm lifestyle.
Biofilm formation is a conserved strategy for long-term bacterial survival in nature and during infections. Biofilms are multicellular aggregates of cells enmeshed in an extracellular matrix. The RetS, GacS and LadS sensors control the switch from a planktonic to a biofilm mode of growth in Pseudomonas aeruginosa. Here we detail our approach to identify environmental triggers of biofilm formation by investigating environmental conditions that repress expression of the biofilm repressor RetS. Mg(2+) limitation repressed the expression of retS leading to increased aggregation, exopolysaccharide (EPS) production and biofilm formation. Repression of retS expression under Mg(2+) limitation corresponded with induced expression of the GacA-controlled small regulatory RNAs rsmZ and rsmY and the EPS biosynthesis operons pel and psl. We recently demonstrated that extracellular DNA sequesters Mg(2+) cations and activates the cation-sensing PhoPQ two-component system, which leads to increased antimicrobial peptide resistance in biofilms. Here we show that exogenous DNA and EDTA, through their ability to chelate Mg(2+), promoted biofilm formation. The repression of retS in low Mg(2+) was directly controlled by PhoPQ. PhoP also directly controlled expression of rsmZ but not rsmY suggesting that PhoPQ controls the equilibrium of the small regulatory RNAs and thus fine-tunes the expression of genes in the RetS pathway. In summary, Mg(2+) limitation is a biologically relevant environmental condition and the first bonafide environmental signal identified that results in transcriptional repression of retS and promotes P. aeruginosa biofilm formation.


Current status of orthopaedic microsurgery in Japan. Personal experience and review of the literature.
Among the Japanese contributions in reconstructive microsurgery, transplantation of composite tissues and replantation of amputated extremities are the most representative. With the standardization of the techniques of microsurgery and replantation, these operations have become relatively common. By July 1981, 3126 replantations had been performed in 78 hospitals with an 80%-90% success rate. Recently, surgeons have focused on the recovery of function. A recent analysis of the author's series of replantations through the end of 1980, using a scoring system of function evaluation criteria, showed that results in 72% of the 181 patients involving 259 upper extremity replants were excellent or good.


Assessment of Driver's Stress using Multimodal Biosignals and Regularized Deep Kernel Learning.
In this work, we classify the stress state of car drivers using multimodal physiological signals and regularized deep kernel learning. Using a driving simulator in a controlled environment, we acquire electrocardiography (ECG), electrodermal activity (EDA), photoplethysmography (PPG), and respiration rate (RESP) from N = 10 healthy drivers in experiments of 25min duration with different stress states (5min resting, 10min driving, 10min driving + answering cognitive questions). We manually remove unusable segments and approximately 4h of data remain. Multimodal time and frequency features are extracted and employed to regularized deep kernel machine learning based on a fusion framework. Task-specific representations of different physiological signals are combined using intermediate fusion. Subsequently, the fused multimodal features are fed a support vector machine (SVM) and a random forest (RF) for stress classification. The experimental results show that the proposed approach can discriminate between stress states. The combination of PPG and ECG using RF as classifier yields the highest F1-score of 0.97 in the test set. PPG only and RF yield a maximum F1-score of 0.90. Furthermore, subject-specific cross-validation improves performance. ECG and PPG signals are reliable in classifying the stress state of a car driver. In summary, the proposed framework could be extended to real-time stress state assessment in driving conditions.


Measurement of nitric oxide in hippocampal slices: induction with nitroso compounds and the effect of depolarization.
Nitric oxide not only acts as a messenger for different physiological processes, but also mediates neurotoxicity associated with a variety of neurological disorders including epilepsy. The molecular mechanisms behind these actions are unclear. In this study, we aimed to detect relative amounts of NO released from rat hippocampal slices by chemiluminescence measurements under NMDA stimulation and spontaneous depolarization conditions. Hippocampal slices were preferred because of their functional integrity useful in simulating in vivo conditions. The reliability of the system was verified by administering increasing concentrations of a NO donor sodium nitroprusside in different redox milieu and a NO scavenger, carboxy-2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (carboxy PTIO). The redox versatility of NO allows interconversion from neuroprotective to neurotoxic species by a change in the ambient redox milieu. We have quantitated NO formed under NMDA stimulation and spontaneous depolarization conditions, and showed that depolarization increased NO formation and was excitotoxic for the neural tissue.