Theoretical study of the hydrolysis mechanism of 2-pyrone-4,6-dicarboxylate (PDC) catalyzed by LigI.
2-Pyrone-4,6-dicarboxylate lactonase (LigI) is the first identified enzyme from amidohydrolase superfamily that does not require a divalent metal ion for catalytic activity. It catalyzes the reversible hydrolysis of 2-pyrone-4,6-dicarboxylate (PDC) to 4-oxalomesaconate (OMA) and 4-carboxy-2-hydroxymuconate (CHM) in the degradation of lignin. In this paper, a combined quantum mechanics and molecule mechanics (QM/MM) approach was employed to study the reaction mechanism of LigI from Sphingomonas paucimobilis. According to the results of our calculations, the whole catalytic reaction contains three elementary steps, including the nucleophilic attack, the cleavage of CO of lactone (substrate) and the intramolecular proton transfer. The intermediate has two intramolecular proton transfer pathways, due to which, two final hydrolysis products can be obtained. The energy profile indicates that 4-carboxy-2-hydroxymuconate (CHM) is the main hydrolysis product, therefore, the isomerization between 4-carboxy-2-hydroxymuconate (CHM) and 4-oxalomesaconate (OMA) is suggested to occur in solvent. During the catalytic reaction, residue Asp248 acts as a general base to activate the hydrolytic water molecule. Although His31, His33 and His180 do not directly participate in the chemical process, they play assistant roles by forming electrostatic interactions with the substrate and its involved species in activating the carbonyl group of the substrate and stabilizing the intermediates and transition states.


Rab6 dependent post-Golgi trafficking of HSV1 envelope proteins to sites of virus envelopment.
Herpes simplex virus 1 (HSV1) is an enveloped virus that uses undefined transport carriers for trafficking of its glycoproteins to envelopment sites. Screening of an siRNA library against 60 Rab GTPases revealed Rab6 as the principal Rab involved in HSV1 infection, with its depletion preventing Golgi-to-plasma membrane transport of HSV1 glycoproteins in a pathway used by several integral membrane proteins but not the luminal secreted protein Gaussia luciferase. Knockdown of Rab6 reduced virus yield to 1% and inhibited capsid envelopment, revealing glycoprotein exocytosis as a prerequisite for morphogenesis. Rab6-dependent virus production did not require the effectors myosin-II, bicaudal-D, dynactin-1 or rabkinesin-6, but was facilitated by ERC1, a factor involved in linking microtubules to the cell cortex. Tubulation and exocytosis of Rab6-positive, glycoprotein-containing membranes from the Golgi was substantially augmented by infection, resulting in enhanced and targeted delivery to cell tips. This reveals HSV1 morphogenesis as one of the first biological processes shown to be dependent on the exocytic activity of Rab6.


Belgian Consensus Recommendations to Prevent Vitamin K Deficiency Bleeding in the Term and Preterm Infant.
Neonatal vitamin K prophylaxis is essential to prevent vitamin K deficiency bleeding (VKDB) with a clear benefit compared to placebo. Various routes (intramuscular (IM), oral, intravenous (IV)) and dosing regimens were explored. A literature review was conducted to compare vitamin K regimens on VKDB incidence. Simultaneously, information on practices was collected from Belgian pediatric and neonatal departments. Based on the review and these practices, a consensus was developed and voted on by all co-authors and heads of pediatric departments. Today, practices vary. In line with literature, the advised prophylactic regimen is 1 or 2 mg IM vitamin K once at birth. In the case of parental refusal, healthcare providers should inform parents of the slightly inferior alternative (2 mg oral vitamin K at birth, followed by 1 or 2 mg oral weekly for 3 months when breastfed). We recommend 1 mg IM in preterm <32 weeks, and the same alternative in the case of parental refusal. When IM is perceived impossible in preterm <32 weeks, 0.5 mg IV once is recommended, with a single additional IM 1 mg dose when IV lipids are discontinued. This recommendation is a step towards harmonizing vitamin K prophylaxis in all newborns.


Trichinellosis survey in the wild boar from the Toledo mountains in south-western Spain (2007-2008): molecular characterization of Trichinella isolates by ISSR-PCR.
In Spain, trichinellosis represents a public health problem, with an average of five outbreaks per year, wild boar meat being the main source of infection. A trichinellosis survey (2007-2008 hunting campaign) was carried out on wild boars in the Toledo Mountains (south-western Spain, EU) in the context of a surveillance programme on wildlife diseases. A total of 2216 wild boars from different locations of the region were examined. The examination was carried out by veterinarians in the local abattoir (Matadero Municipal de Toledo). The positive samples were sent to the Department of Parasitology (Facultad de Farmacia, UCM) for experimental isolation and specific identification by inter-simple sequence repeat-polymerase chain reaction (ISSR-PCR). Using this technique we identified 17 isolates as Trichinella spiralis with an electrophoretic profile indistinguishable from the T. spiralis reference strain (ISS48). We confirmed that ISSR-PCR is a robust technique for the molecular identification of Trichinella isolates. According to our results, the prevalence of T. spiralis in wild boars from the Toledo Mountains (>800 m above sea level) during the hunting season was approximately 0.77%. The prevalence of T. spiralis (100% of our observations) is a good example of the persistence of this species in sylvatic conditions (coming from the domestic cycle), if a good wild host is abundant. Our observations confirm the major prevalence of T. spiralis over T. britovi in this region, as well as the risk to human health represented by the consumption of uninspected wild boar meat.


Psychological stress has no association with salivary levels of β-defensin 2 and β-defensin 3.
Recent studies suggest that stress can predispose an individual to the development of periodontal disease, but the exact biological mechanism is unknown. Considering that psychological stress can down-regulate the production of β-defensins (antimicrobial peptides produced in the oral cavity), the aim of the present study was to evaluate the association between stress and salivary levels of β-defensin 2 (HBD-2) and β-defensin 3 (HBD-3). For this purpose, seventy five volunteers, classified as periodontally healthy, were submitted to a psychological evaluation using a validated questionnaire (Questionnaire of Lipp-ISS). Following analysis of the questionnaires, the subjects were divided in two groups (Group A: Absence of stress and Group B: Presence of stress). Unstimulated saliva samples were collected and the concentration of total protein was determined using the BCA method, and the concentrations of HBD-2 and HBD-3 were determined by ELISA. The levels of total protein did not show a statistically significant difference between the groups. Analyses of HBD-2 and HBD-3 concentrations indicate that the stress condition was not associated with the levels of either peptide in saliva (P=0.3664 for HBD-2 and P=0.3608 for HBD-3). In periodontally healthy subjects, HBD-2 and HBD-3 levels are not influenced by stress.


PST-PRNA: prediction of RNA-binding sites using protein surface topography and deep learning.
Protein-RNA interactions play essential roles in many biological processes, including pre-mRNA processing, post-transcriptional gene regulation and RNA degradation. Accurate identification of binding sites on RNA-binding proteins (RBPs) is important for functional annotation and site-directed mutagenesis. Experimental assays to sparse RBPs are precise and convincing but also costly and time consuming. Therefore, flexible and reliable computational methods are required to recognize RNA-binding residues. In this work, we propose PST-PRNA, a novel model for predicting RNA-binding sites (PRNA) based on protein surface topography (PST). Taking full advantage of the 3D structural information of protein, PST-PRNA creates representative topography images of the entire protein surface by mapping it onto a unit spherical surface. Four kinds of descriptors are encoded to represent residues on the surface. Then, the potential features are integrated and optimized by using deep learning models. We compile a comprehensive non-redundant RBP dataset to train and test PST-PRNA using 10-fold cross-validation. Numerous experiments demonstrate PST-PRNA learns successfully the latent structural information of protein surface. On the non-redundant dataset with sequence identity of 0.3, PST-PRNA achieves area under the receiver operating characteristic curves (AUC) value of 0.860 and Matthew's correlation coefficient value of 0.420. Furthermore, we construct a completely independent test dataset for justification and comparison. PST-PRNA achieves AUC value of 0.913 on the independent dataset, which is superior to the other state-of-the-art methods. The code and data are available at https://www.github.com/zpliulab/PST-PRNA. A web server is freely available at http://www.zpliulab.cn/PSTPRNA. Supplementary data are available at Bioinformatics online.


99mTc-labeled antihuman epidermal growth factor receptor antibody in patients with tumors of epithelial origin: Part III. Clinical trials safety and diagnostic efficacy.
Monoclonal antibody (moAb) ior egf/r3 is an IgG2a that recognizes the epidermal growth factor receptor (EGF-R). The aim of this study was to evaluate the diagnostic efficacy of the 99mTc-labeled moAb ior egf/r3 for the detection of epithelial-derived tumors, their metastases and recurrences. One hundred forty-eight adult patients (51 women, 97 men; mean age 53 +/- 13 y) who were suspected of having cancer of epithelial origin were administered 3 mg/50 mCi (1.85 GBq) 99mTc-labeled moAb ior egf/r3 by intravenous bolus injection. Planar anterior and posterior images of the lesion sites and suspected metastases were acquired at 2, 4, 6 and 24 h after injection, and SPECT images were scanned at 5 h postinjection, using a 360 degrees circular orbit with 64 images. The backprojection method was used for image reconstruction with a Hamming-Hann filter. Labeling efficiency was always greater than 98.5% +/- 2.1 %. No adverse reactions or side effects were observed. Results of the biopsy specimens showed that 85.1% (126/148) of the patients had tumors of epithelial origin, 14.2% (21/148) were negative and 0.7% (1/148) had non-Hodgkin's lymphoma. The sensitivity rate by organ was as follows: brain (8/8, 100%), digestive tract (10/11, 90.9%), head and neck (17/23, 73.9%), lung (52/62, 83.9%) and breast (16/18, 88.9%). Overall sensitivity, specificity, accuracy, and positive and negative predictive values of the immunoscintigraphic imaging were 84.2% (106/126), 100.0% (22/22), 86.5% (128/148), 100% (106/106) and 52.4% (22/42), respectively. New metastases not identified previously by other diagnostic methods were detected in the 50% of the patients. Immunoscintigraphy with 99mTc-labeled moAb ior egf/r3 could be a useful procedure for the diagnosis and follow-up of the patients with tumors of epithelial origin.


An ab initio molecular orbital analysis of phosphorothioate mono-anion.
Ab initio calculations at the STO-3G level show that the energy of the highest occupied molecular orbital (HOMO) of the di-protonated phosphorothioate mono-anion is 0.0421 a.u. higher than that of the HOMO of the corresponding phosphate mono-anion. The electron density in the phosphorothioate HOMO is highly focused on the sulfur atom. The results of this ab initio molecular orbital analysis are consistent with the model of the phosphorothioate sulfur as a soft nucleophile. The ab initio molecular orbital analysis presented in this paper will be useful for predicting and rationalizing the probability of reaction of phosphorothioate anions with electrophiles and helps to provide a basis for the rational design of phosphorothioate-based oligonucleotide drugs.


Density estimation of Yangtze finless porpoises using passive acoustic sensors and automated click train detection.
A method is presented to estimate the density of finless porpoises using stationed passive acoustic monitoring. The number of click trains detected by stereo acoustic data loggers (A-tag) was converted to an estimate of the density of porpoises. First, an automated off-line filter was developed to detect a click train among noise, and the detection and false-alarm rates were calculated. Second, a density estimation model was proposed. The cue-production rate was measured by biologging experiments. The probability of detecting a cue and the area size were calculated from the source level, beam patterns, and a sound-propagation model. The effect of group size on the cue-detection rate was examined. Third, the proposed model was applied to estimate the density of finless porpoises at four locations from the Yangtze River to the inside of Poyang Lake. The estimated mean density of porpoises in a day decreased from the main stream to the lake. Long-term monitoring during 466 days from June 2007 to May 2009 showed variation in the density 0-4.79. However, the density was fewer than 1 porpoise/km(2) during 94% of the period. These results suggest a potential gap and seasonal migration of the population in the bottleneck of Poyang Lake.


Ictal SPECT in children with epilepsy: comparison with intracranial EEG and relation to postsurgical outcome.
In order to validate the ability of ictal single photon emission computed tomography (SPECT) to localize the epileptogenic zone (EZ) in children, we compared in 20 patients aged from 10 months to 17 years (mean 6.5 years) the topography of the area of increased ictal perfusion (IPA), determined on the basis of ictal minus interictal scan values, with that of the EZ determined by intracranial EEG recordings and assessed its relationship with the postsurgical outcome. Eighteen patients had symptomatic epilepsy and 10 had extratemporal epilepsy. All patients except one had an ictal injection (mean time lag from clinical seizure onset was 18 s). Ictal and interictal SPECT images were successively co-registered, normalized, subtracted, smoothed and superimposed on MRI. All patients with ictal injection exhibited one or several IPAs. The topography of the 'highest' IPA, i.e. the maximal cerebral blood flow (CBF) change between ictal and interictal SPECT, significantly colocalized with the site of onset of the discharge, and that of the lower IPAs with that of the area of propagation (P < 0.0001). At a threshold of 30% of the maximal CBF change, the IPAs detected the onset of the discharge with a sensitivity of 0.80 and a specificity of 0.70. The highest IPA localized the EZ in 12 out of 15 patients. In the three others it missed the EZ and showed the area of propagation because of rapid seizure propagation or of infraclinical seizure onset. Among the patients with favourable surgery outcome, the highest IPA colocalized with the resected area in 70% of cases. Ictal SPECT could therefore plays an important role as a non-invasive presurgical method of investigation by optimizing the placement of intracranial electrodes, thus improving the postsurgery outcome of paediatric partial epilepsy.


Macrophage-tumour cell interactions: identification of MUC1 on breast cancer cells as a potential counter-receptor for the macrophage-restricted receptor, sialoadhesin.
In many carcinomas, infiltrating macrophages are commonly found closely associated with tumour cells but little is known concerning the nature or significance of adhesion molecules involved in these cellular interactions. Here we demonstrate in primary human breast cancers that sialoadhesin (Sn), a macrophage-restricted adhesion molecule, is frequently expressed on infiltrating cells that often make close contact with breast carcinoma cells. To determine whether Sn could act as a specific receptor for ligands on breast cancer cell lines, binding assays were performed with a recombinant form of the protein fused to the Fc portion of human immunoglobulin G1 (IgG1) (Sn-Fc). Sn-Fc was found to bind specifically and in a sialic acid-dependent manner to the breast cancer cell lines MCF-7, T47.D and BT-20 both in solid- and solution-phase binding assays. To investigate the nature of the sialoglycoproteins recognized by Sn on breast cancer cells, MCF-7 cells were labelled with [6-3H]glucosamine. Following precipitation with Sn-Fc, a major band of approximately 240000 MW was revealed, which was shown in reprecipitation and Western blotting experiments to be the epithelial mucin, MUC1.


Impact of organophosphate exposure on farmers' health in Kulon Progo, Yogyakarta: Perspectives of physical, emotional and social health.
The exposure of organophosphate could be caused by the absorption in some parts of the body like skin and breath. Toxicity may cause nausea, vomiting and dizziness which are not too specific related with the pesticide toxicity. The purpose of this study is to understand the association between organophosphate exposure and farmers' health in Kulon Progo County from the perspectives of physical, emotional and social health. This study was conducted using descriptive observational design. The blood sample was collected during harvesting periods in 2016. The inclusion criterion of farmers was using organophosphate-contained pesticide during the planting period of red onion. The farmers who had renal disease, liver disease and cancer were excluded. The organophosphate exposure parameters were the duration and frequency of pesticide application, width of the area, serum cholinesterase activity and the completeness of personal protective equipment. Among 84 farmers, most of them were male (85.7%), and the mean age was 49.1 (standard deviation: 12.5) years; 71.4% of the subjects experienced tremor, 17.86% experienced dizziness and 8.33% subjects experienced nausea-vomiting after pesticide application. According to the pesticide application, in average, subjects used pesticide 1.4 h/day with the area of 1.285 m2. The frequency of pesticide used is three times per week. Around 97.6% subjects used incomplete personal protective equipment. The average of serum cholinesterase activity in subjects with tremor is higher than subjects without tremor (p > 0.05). There is a significant association between serum cholinesterase activity and creatinine content (p < 0.05). The farmers' quality-of-life domain scores are lower than the scores of the normal population in Yogyakarta. Organophosphate exposure may affect the farmers' physical health and quality of life.


Rationale for a preliminary operational definition of physical frailty and sarcopenia in the SPRINTT trial.
In the present article, the rationale that guided the operationalization of the theoretical concept of physical frailty and sarcopenia (PF&S), the condition of interest for the "Sarcopenia and Physical Frailty in Older People: Multicomponent Treatment Strategies" (SPRINTT) trial, is presented. In particular, the decisions lead to the choice of the adopted instruments, and the reasons for setting the relevant thresholds are explained. In SPRINTT, the concept of physical frailty is translated with a Short Physical Performance Battery score of ≥3 and ≤9. Concurrently, sarcopenia is defined according to the recent definitions of low muscle mass proposed by the Foundation for the National Institutes of Health-Sarcopenia Project. Given the preventive purpose of SPRINTT, older persons with mobility disability (operationalized as incapacity to complete a 400-m walk test within 15 min; primary outcome of the trial) at the baseline are not included within the diagnostic spectrum of PF&S.


The fibroblast mitogenic activity released from human basophilic cell line KU812 is separate from tryptase and PDGF expression.
The human leukaemia cell line KU812 has previously been used to study basophil differentiation. In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts. KU812 cells were treated with tetradecanoyl-phorbol-13-acetate (TPA), conditioned medium from the human T-cell line Mo (Mo-CM), or cultured under serum free conditions. After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity. Mo-CM and serum starvation increased the expression while TPA treatment down-regulated the expression of Fc epsilon RI-alpha chain. An increase in tryptase content in cell extracts was detected after 4 days of culture in serum-free medium or in the presence of Mo-CM. KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA. Mast cell-derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells. Furthermore, affinity-purified lung tryptase did not show any mitogenic activity. Platelet-derived growth factor was also excluded. Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.


Benefits of combined exercise training on arterial stiffness and blood pressure in spontaneously hypertensive rats treated or not with dexamethasone.
Dexamethasone (DEX)-induced arterial stiffness is an important side-effect, associated with hypertension and future cardiovascular events, which can be counteracted by exercise training. The aim of this study was to evaluate the mechanisms induced by combined training to attenuate arterial stiffness and hypertension in spontaneously hypertensive rats treated or not with dexamethasone. Spontaneously hypertensive rats (SHR) underwent combined training for 74 days and were treated with dexamethasone (50 µg/kg s. c.) or saline solution during the last 14 days. Wistar rats were used as controls. Echocardiographic parameters, blood pressure (BP) and pulse wave velocity (PWV), as well as histological analyses of the heart and aorta, carotid and femoral arteries were performed. At the beginning, SHR had higher BP and PWV compared with Wistar rats. After 60 days, while BP increased in sedentary SHR, combined exercise training decreased BP and PWV. After 74d, the higher BP and PWV of sedentary SHR was accompanied by autonomic imbalance to the heart, cardiac remodeling, and higher arterial collagen deposition. DEX treatment did not change these parameters. On the other hand, trained SHR had reduced BP and PWV, which was associated with better autonomic balance to the heart, reduced myocardial collagen deposition, as well as lower arterial collagen deposition. The results of this study suggest that combined training, through the reduction of aortic collagen deposition, is an important strategy to reduce arterial stiffness in spontaneously hypertensive rats, and these lower responses were maintained regardless of dexamethasone treatment.


Hog barn dust extract stimulates IL-8 and IL-6 release in human bronchial epithelial cells via PKC activation.
Hog barn workers have an increased incidence of respiratory tract symptoms and demonstrate an increase in lung inflammatory mediators, including interleukin (IL)-8 and IL-6. Utilizing direct kinase assays for protein kinase C (PKC) activation, we demonstrated that dust from hog confinement facilities, or hog dust extract (HDE), augments PKC activity of human airway epithelial cells in vitro. A 5% dilution of HDE typically stimulates an approximately twofold increase in human bronchial epithelial cell (HBEC) PKC activity compared with control medium-treated cells. This increase in PKC is observed with 15 min of HDE treatment, and kinase activity reaches peak activity by 1-2 h of HDE treatment before returning to baseline PKC levels between 6 and 24 h. The classic PKC inhibitor, calphostin C, blocks HDE-stimulated PKC activity and associated IL-8 and IL-6 release. Desensitization to HDE stimulation of PKC activation does not appear to occur because subsequent exposures to HDE after an initial exposure result in further augmentation of PKC. Detoxification of HDE with polymyxin B to remove endotoxin did not change PKC activation or IL-8 release, suggesting that endotoxin is not solely responsible for HDE augmentation of PKC. These data support the hypothesis that HDE exposure augments HBEC IL-8 and IL-6 release via a PKC-dependent pathway.


[Treatment of malignant intracranial germ-cell tumors in children. 7 cases].
Seven children, aged from 9 to 16 years, with intracranial germ cell tumour received post-operative chemotherapy prior to radiotherapy. Treatment consisted of several courses of vinblastine, bleomycin and cisplatin alternating or not with courses of cyclophosphamide and actinomycin D. In the 6 children whose tumours secreted alphafoetoprotein or chorionic gonadotrophin, chemotherapy brought these markers down to normal. Tumoral regression at radiology exceeded 75 per cent in 6 patients. These results demonstrate the effectiveness of chemotherapy against intracranial germ cell tumours such as pure germinomas or secreting tumours. They encourage the systematic use of pre-irradiation chemotherapy in order to reduce the radiation doses delivered and to improve the prognosis.


Sequential changes in the lipids of developing proplastids isolated from green maize leaves.
Changes in lipid composition were followed as a proplastid develops into a chloroplast. Methods were devised for the isolation of developing proplastids from sections of five different ages from the same 7-day-old maize (Zea mays var. Kelvedon Glory) leaf. Electron micrographs illustrate the homogeneity of the five types of plastid suspension, minimal contamination with other cytoplasmic membranes, and the presence of morphologically intact plastids in the proportions 85% (youngest), 85%, 80%, 70% and 60% (oldest), respectively. Both bundle sheath and mesophyll plastids are well preserved in isolation. Plastid numbers were determined from calibration curves of the chlorophyll content of each type of suspension, and lipid values then expressed as nmoles/10(6) plastids. Monogalactosyl diglyceride (MGDG), digalactosyl diglyceride (DGDG), sulfoquinovosyl diglyceride, and phosphatidyl glycerol (PG) all increase during plastid development but the rate of increase is different for each lipid. The largest changes are in MGDG (6-fold) and DGDG (4-fold). Phosphatidyl choline shows a continuous decline during plastid development. Phosphatidyl inositol and phosphatidyl ethanolamine were found in all the suspensions in low concentrations (0.4-4.0% of total lipid): calculations showed their presence could not be accounted for by bacterial or mitochondrial contamination. The increase in PG parallels the chlorophyll changes during development and at maturity 1 molecule of PG is present per 3 molecules of chlorophyll. The results are discussed in the context of the molecular structure of the photosynthetic thylakoid membranes.


Health in a fragile state: a five-year review of mortality patterns and trends at Somalia's Banadir Hospital.
The recurrent civil conflict in Somalia has impeded progress toward improving health and health care, with lack of data and poor performance of health indicators. This study aimed at making inference about Banadir region by exploring morbidity and mortality trends at Banadir Hospital. This is one of the few functional hospitals during war. A retrospective analysis was conducted with data collected at Banadir Hospital for the period of January 2008-December 2012. The data were aggregated from patient records and summarized on a morbidity and mortality surveillance form with respect to age groups and stratified by sex. The main outcome was the number of patients that died in the hospital. Chi-square tests were used to evaluate the association between sex and hospital mortality. Conditions of infectious origin were the major presentations at the hospital. The year 2011 recorded the highest number of cases of diarrhea and mortality due to diarrhea. The stillbirth rate declined during the study period from 272 to 48 stillbirths per 1,000 live births by 2012. The sum of total cases that were attended to at the hospital by the end of 2012 was four times the number at the baseline year of the study in 2008; however, the overall mortality rate among those admitted declined between 2008 and 2012. There was reduction in patient mortality at the hospital over the study period. Data from Banadir Hospital are consistent with findings from Banadir region and could give credible public health reflections for the region given the lack of data on a population level.


Non-Arteritic Anterior Ischaemic Optic Neuropathy with Progressive Macular Ganglion Cell Atrophy due to COVID-19.
A 72-year-old man with type II diabetes mellitus presented with sudden painless vision loss and an inferior visual field defect in his right eye. He had previously tested positive for COVID-19 disease with the symptoms starting 13 days before the onset of vision loss. His decimal visual acuity, 55 days after the onset of visual symptoms, was 0.3 and this decreased over the following week to counting fingers. 24-2 visual field analysis revealed an inferior altitudinal defect. Dilated fundus examination revealed mild optic disc swelling in the right eye. The left eye was normal. He was diagnosed with non-artertic anterior ischaemic optic neuropathy (NAION). On spectral domain optical coherence tomography there was retinal thinning in the supero-temporal foveal area. Macular ganglion cell layer - inner plexiform retinal layer complex analysis showed progressive atrophy that developed from the supero-temporal to the infero-nasal fovea. COVID-19 infection may lead to NAION.


Biological Landscape of Triple Negative Breast Cancers Expressing CTLA-4.
Patients with triple-negative breast cancer (TNBC) have a poor prognosis, partly because of the absence of targeted therapies. Recognition of the key role of immune responses against cancer has allowed the advent of immunotherapy, focused on the inhibition of negative immune checkpoints, such as CTLA-4. CTLA-4 is also expressed in some cancer cells, but its activity in tumor cells is not completely understood. Thus, the aim of the present work was to determine the biological landscape and functions of CTLA-4 expressed in TNBC cells through preclinical and in silico analysis. Exploration of CTLA-4 by immunohistochemistry in 50 TNBC tumors revealed membrane and cytoplasmic expression at different intensities. Preclinical experiments, using TNBC cell lines, showed that stimulation of CTLA-4 with CD80 enhances activation of the ERK1/2 signaling pathway, while CTLA-4 blockade by Ipilimumab induces the activation of AKT and reduces cell proliferation in vitro. We then developed an analytic pipeline to define the effects of CTLA-4 in available public data that allowed us to identify four distinct tumor clusters associated with CTLA-4 activation, which are characterized by enrichment of distinctive pathways associated with cell adhesion, MAPK signaling, TGF-ß, VEGF, TNF-α, drug metabolism, ion and amino acid transport, and KRAS signaling, among others. In addition, blockade of CTLA-4 induced increased secretion of IL-2 by tumor cells, suggesting that the receptor regulates cellular functions that may impact the immune microenvironment. This is relevant because a deep characterization of immune infiltrate, conducted using public data to estimate the abundancies of immune-cell types, showed that CTLA-4-activated-like tumors present a conditional immune state similar to an escape phenotype exploited by cancer cells. Finally, by interrogating transcriptional predictors of immunotherapy response, we defined that CTLA-4 activation correlates with high immune scores related to good clinical predicted responses to anti-CTLA-4 therapy. This work sheds new light on the roles of activated CLTA-4 in the tumor compartment and suggests an important interplay between tumor CLTA-4-activated portraits and immune-infiltrating cell populations.


A review of green methods used in starch-polyphenol interactions: physicochemical and digestion aspects.
The interactions of starch with lipids, proteins, and other major food components during food processing are inevitable. These interactions could result in the formation of V-type or non-V-type complexes of starch. The starch-lipid complexes have been intensively studied for over five decades, however, the complexes of starch and polyphenols are relatively less studied and are the subject of recent interest. The interactions of starch with polyphenols can affect the physicochemical properties and its digestibility. The literature has highlighted several green methods such as ultrasound, microwave, high pressure, extrusion, ball-milling, cold plasma etc., to assist interactions of starch with polyphenols. However, comprehensive information on green methods to induce starch-polyphenol interactions is still scarce. Therefore, in light of the importance and potential of starch-polyphenol complexes in developing functional foods with low digestion, this review has summarized the novel green methods employed in interactions of starch with flavonoids, phenolic acids and tannins. It has been speculated that flavonoids, phenolic acids, and tannins, among other types of polyphenols, may have anti-digestive activities and are also revealed for their interaction with starch to form either an inclusion or non-inclusion complex. Further information on the effects of these interactions on physicochemical parameters to understand the chemistry and structure of the complexes is also provided.


Selective attention and search images in pigeons.
In a visual search task, pigeons detected targets when pretrial visual cues or blocked trial sequences signaled the target's identity. Sequential priming was robust over a wide range of intertrial intervals, but visual priming was unstable when the delay between cue offset and display onset was varied. Larger target set sizes enhanced sequential, but not visual, priming. Sequential priming did not depend on display size over the range of relatively large displays used. However, ambiguously cued targets in small displays were detected more quickly than primed targets in large displays. These findings suggest that naturalistic selection biases, or "search images," may be attributable to sequential priming and that the common attentional mechanism has moderately selective properties.


Assessment of nitrogen and phosphorus flows in agricultural and urban systems in a small island under limited data availability.
Nitrogen (N) and phosphorus (P) are two essential macronutrients required in agricultural production. The major share of this production relies on chemical fertilizer that requires energy and relies on limited resources (P). Since these nutrients are lost to the environment, there is a need to shift from this linear urban metabolism to a circular metabolism in which N and P from domestic waste and wastewater are reused in agriculture. A first step to facilitate a transition to more circular urban N and P management is to understand the flows of these resources in a coupled urban-agricultural system. For the first time this paper presents a Substance Flow Analysis (SFA) approach for the assessment of the coupled agricultural and urban systems under limited data availability in a small island. The developed SFA approach is used to identify intervention points that can provide N and P stocks for agricultural production. The island of St. Eustatius, a small island in the Caribbean, was used as a case study. The model developed in this study consists of eight sub-systems: agricultural and natural lands, urban lands, crop production, animal production, market, household consumption, soakage pit and open-dump landfill. A total of 26 flows were identified and quantified for a period of one year (2013). The results showed that the agricultural system is a significant source for N and P loss because of erosion/run-off and leaching. Moreover, urban sanitation systems contribute to deterioration of the island's ecosystem through N and P losses from domestic waste and wastewater by leaching and atmospheric emission. Proposed interventions are the treatment of blackwater and greywater for the recovery of N and P. In conclusion, this study allows for identification of potential N and P losses and proposes mitigation measures to improve nutrient management in a small island context.


HEALTH ASSESSMENT OF WILD LOWLAND TAPIRS (TAPIRUS TERRESTRIS) IN THE HIGHLY THREATENED CERRADO BIOME, BRAZIL.
Over 2 yr, we assessed the health of 35 lowland tapirs (Tapirus terrestris) in the Brazilian Cerrado (CE) biome, an area that is highly affected by human activities. This involved physical examinations, hematology and blood biochemistry, urinalysis, fecal parasitologic evaluation, microbial profiling of anatomic cavities and lesions, and serologic surveys for evidence of infectious agents. Research methods closely resembled those used in previous tapir health assessments in the Atlantic Forest (AF) and Pantanal (PA) biomes, allowing for a comparison among the three populations. Although not reaching statistical significance (P>0.05), tapirs from the CE exhibited poorer body and skin condition as compared to animals from the AF and PA. Furthermore, there were higher prevalences of dental problems and traumatic lesions as compared to those from the AF and PA. Eight of the 12 hematologic parameters evaluated and 17 of the 30 biochemical parameters differed significantly (P<0.05) between the tapirs from CE and those from the AF and PA. We isolated 24 different microbiologic strains from swabs of anatomic cavities and dermal lesions, of which five taxa had not previously been found in the AF or PA. We detected serum antibodies to Leptospira interrogans, bluetongue virus, and porcine parvovirus. Overall, our results suggested that tapirs from the CE exhibited more health abnormalities than tapirs in the AF and PA, possibly due to a greater exposure to environmental disturbances in the area.


PREVALENCE OF POLYPOIDAL CHOROIDAL VASCULOPATHY IN WHITE PATIENTS WITH EXUDATIVE AGE-RELATED MACULAR DEGENERATION: Systematic Review and Meta-Analysis.
Polypoidal choroidal vasculopathy (PCV) is a disease with significant inter-ethnical differences. In this study, we systematically review the literature on the prevalence of PCV in whites referred with a diagnosis of exudative age-related macular degeneration (AMD). We searched PubMed, Embase, the Cochrane Library, and the Web of Science on 24 March, 2017 for studies evaluating the prevalence of PCV in white patients with exudative AMD. Data extraction and risk of bias assessments were performed in duplicate. Studies were included for a qualitative review and a meta-analysis, including subgroup analysis for differences in age and sex. We included data from 11 studies (>2,200 participants). For diagnosis, indocyanine green angiography was used together with a set of supporting criteria on fundus examination and optical coherence tomography. Extramacular location was more prevalent in eyes with PCV. Drusen was present in the fellow eye in 17% to 27%. Pooled prevalence of PCV in white patients with exudative AMD was 8.7% (confidence interval 95%: 7.2%-10.3%). Patients with PCV were 3.7 years (confidence interval 95%: 2.1 years-5.3 years) younger than those with other exudative AMD. Sex did not differ significantly. Polypoidal choroidal vasculopathy is not a rare subtype of exudative AMD in whites-it is present in approximately one in 11 patients.


Asymmetric Notch activity by differential inheritance of lysosomes in human neural stem cells.
Symmetric and asymmetric cell divisions are conserved strategies for stem cell expansion and the generation of more committed progeny, respectively. Here, we demonstrate that in human neural stem cells (NSCs), lysosomes are asymmetrically inherited during mitosis. We show that lysosomes contain Notch receptors and that Notch activation occurs the acidic lysosome environment. The lysosome asymmetry correlates with the expression of the Notch target gene HES1 and the activity of Notch signaling in the daughter cells. Furthermore, an asymmetry of lysosomes and Notch receptors was also observed in a human organoid model of brain development with mitotic figures showing preferential inheritance of lysosomes and Notch receptor in that daughter cell remaining attached to the apical membrane. Thus, this study suggests a previously unknown function of lysosomes as a signaling hub to establish a bias in Notch signaling activity between daughter cells after an asymmetric cell division of human NSCs.


Enhancement by Metallic Tube Filling of the Mechanical Properties of Electromagnetic Wave Absorbent Polymethacrylimide Foam.
By the addition of a carbon-based electromagnetic absorbing agent during the foaming process, a novel electromagnetic absorbent polymethacrylimide (PMI) foam was obtained. The proposed foam exhibits excellent electromagnetic wave-absorbing properties, with absorptivity exceeding 85% at a large frequency range of 4.9⁻18 GHz. However, its poor mechanical properties would limit its application in load-carrying structures. In the present study, a novel enhancement approach is proposed by inserting metallic tubes into pre-perforated holes of PMI foam blocks. The mechanical properties of the tube-enhanced PMI foams were studied experimentally under compressive loading conditions. The elastic modulus, compressive strength, energy absorption per unit volume, and energy absorption per unit mass were increased by 127.9%, 133.8%, 54.2%, and 46.4%, respectively, by the metallic tube filling, and the density increased only by 5.3%. The failure mechanism of the foams was also explored. We found that the weaker interfaces between the foam and the electromagnetic absorbing agent induced crack initiation and subsequent collapses, which destroyed the structural integrity. The excellent mechanical and electromagnetic absorbing properties make the novel structure much more competitive in electromagnetic wave stealth applications, while acting simultaneously as load-carrying structures.


Cell Wall-Treated Lactococcus lactis Increases the Plasmid Transfer Efficiency of Internal Ribosome Entry Site-Incorporated Lactococcal Bicistronic Vector into DF1 Cells.
This study demonstrates that cell wall treatment of Lactococcus lactis harbouring the internal ribosome entry site-incorporated lactococcal bicistronic vector pNZ:VIG mediated the delivery of genes into an eukaryotic cell line, DF1 cells, through bactofection. Bactofection analysis showed that the pNZ:VIG plasmid in L. lactis can be transferred into DF1 cells and that both the VP2 and gfp genes cloned in the plasmid can be transcribed and translated. The protein band relative to the Mr of VP2 protein (49 kDa) was successfully detected via Western blot analysis, while green fluorescence was successfully detected using a fluorescence microscope. The intensity of the bands detected increased for samples treated with both 1.5% (w/v) glycine and 10 μg/mL of lysozyme when compared to L. lactis treated with glycine alone and without treatment. Cell wall treatment of L. lactis with a combination of both glycine and lysozyme was not only shown to mediate plasmid transfer to DF1 cells, but also to increase the plasmid transfer efficiency.


Antiarrhythmic drug therapy. Recent advances and current status.
A number of conventional and newer antiarrhythmic agents are available for the treatment and prophylaxis of ventricular tachycardia and sudden death. Using a multifaceted approach of programmed electrical stimulation studies, drug level determinations, exercise tolerance testing, and 24-hour ambulatory electrocardiographic monitoring, the physician can identify those patients who require therapy and then predict the likelihood of efficacy with each antiarrhythmic agent. This approach affords evaluation of both aspects of the sudden death equation-ectopy frequency (triggering mechanism) and vulnerability to development of sustained ventricular tachycardia (substrate). After institution of therapy, careful follow-up is necessary to document sustained drug efficacy and detect side effects. Serious adverse reactions necessitate a change in antiarrhythmic therapy, as opposed to lowering drug dosage to an ineffective level. The unacceptably high incidence of sudden death due to electrical instability can be reversed only by a rigorous and dedicated long-term approach to the management of serious ventricular arrhythmias.


Renal interstitial hydrostatic pressure and sodium excretion during acute volume expansion in diabetic rats.
Experiments were performed to test the hypothesis that the renal interstitial hydrostatic pressure (RIHP) response to acute volume expansion is suppressed in diabetes mellitus. Sprague-Dawley rats received streptozotocin (STZ rats; 65 mg/kg ip) or vehicle (Sham rats). Two weeks later, RIHP and Na(+) excretion responses to acute graded volume expansion with isotonic saline were quantified under Inactin anesthesia (0.1 mg/kg ip). In Sham rats, acute graded volume expansion to 10% body wt produced increases in RIHP (Delta = 12.2 +/- 2.4 mmHg), urine flow (Delta = 54 +/- 8 microliter. min(-1). g(-1)), and Na(+) excretion (Delta = 11.5 +/- 1.9 mueq. min(-1). g(-1)). In STZ rats, these volume expansion-induced responses were significantly blunted (RIHP by 50%, urine flow by 81%, and Na(+) excretion by 76%). Renal decapsulation eliminated the differences between STZ and Sham rats with regard to volume expansion-induced increases in RIHP, urine flow, and Na(+) excretion. Renal denervation normalized the RIHP response to volume expansion and improved the diuretic and natriuretic responses in STZ rats. Moreover, diuretic and natriuretic responses to direct changes in RIHP (induced by renal interstitial volume expansion) were blunted in STZ rats. We conclude that diminished alterations in RIHP, as well as a reduced impact of RIHP on Na(+) excretion, contribute to the impaired diuretic and natriuretic responses to acute volume expansion during the early stage of diabetes.


Sustained G-->A hypermutation during reverse transcription of an entire human immunodeficiency virus type 1 strain Vau group O genome.
Two full-length human immunodeficiency virus type 1 O sequences are described, one of which was hypermutated in all regions of the genome. This indicates that the intracellular [dTTP]/[dCTP] bias conducive to G-->A hypermutation may be sustained throughout the synthesis of minus-strand DNA. In turn, this suggests the possibility of mutation of host sequences.


Practical total synthesis of ciguatoxin CTX3C by improved protective group strategy.
[structure: see text] Ciguatoxin CTX3C is a representative congener of ciguatoxins, which are known to be the principal causative agents of ciguatera seafood poisoning. The structure of CTX3C spans more than 3 nm and is characterized by 13 ether rings. In this paper, an improved total synthesis of CTX3C is reported. Alteration of the protective group from benzyl ether to 2-naphthylmethyl (NAP) ether drastically increases the yield for final global deprotection and has provided a sufficient amount of sample for further biological studies.


Oropharyngeal colonization with aerobic gram-negative bacilli in respiratory therapists: period prevalence.
During a 3-month winter period, swab pharyngeal cultures for aerobic gram-negative bacilli were obtained weekly for 3 weeks from 44 hospital-based respiratory therapists and 53 nontherapist control hospital personnel. The swab cultures were inoculated directly onto MacConkey agar plates. Aerobic gram-negative bacilli were isolated from 9.1% of the respiratory therapists and from 9.4% of the control subjects. One respiratory therapist was colonized with the same organism on all three occasions, whereas all other colonized subjects were only transiently colonized. Thus, despite frequent exposure to patients and aerosols likely to contain aerobic gram-negative bacilli, respiratory therapists do no appear to have higher pharyngeal colonization rates than other hospital personnel.


Inhibition of deubiquitination by PR-619 induces apoptosis and autophagy via ubi-protein aggregation-activated ER stress in oesophageal squamous cell carcinoma.
Targeting the deubiquitinases (DUBs) has become a promising avenue for anti-cancer drug development. However, the effect and mechanism of pan-DUB inhibitor, PR-619, on oesophageal squamous cell carcinoma (ESCC) cells remain to be investigated. The effect of PR-619 on ESCC cell growth and cell cycle was evaluated by CCK-8 and PI staining. Annexin V-FITC/PI double staining was performed to detect apoptosis. LC3 immunofluorescence and acridine orange staining were applied to examine autophagy. Intercellular Ca2+ concentration was monitored by Fluo-3AM fluorescence. The accumulation of ubi-proteins and the expression of the endoplasmic reticulum (ER) stress-related protein and CaMKKβ-AMPK signalling were determined by immunoblotting. PR-619 could inhibit ESCC cell growth and induce G2/M cell cycle arrest by downregulating cyclin B1 and upregulating p21. Meanwhile, PR-619 led to the accumulation of ubiquitylated proteins, induced ER stress and triggered apoptosis by the ATF4-Noxa axis. Moreover, the ER stress increased cytoplasmic Ca2+ and then stimulated autophagy through Ca2+ -CaMKKβ-AMPK signalling pathway. Ubiquitin E1 inhibitor, PYR-41, could reduce the accumulation of ubi-proteins and alleviate ER stress, G2/M cell cycle arrest, apoptosis and autophagy in PR-619-treated ESCC cells. Furthermore, blocking autophagy by chloroquine or bafilomycin A1 enhanced the cell growth inhibition effect and apoptosis induced by PR-619. Our findings reveal an unrecognized mechanism for the cytotoxic effects of general DUBs inhibitor (PR-619) and imply that targeting DUBs may be a potential anti-ESCC strategy.


Surface Self-Assembly Construction of Therapeutic Contact Lens with Bacterial "Kill-Releasing" and Drug-Reloading Capabilities for Efficient Bacterial Keratitis Treatment.
Bacterial keratitis, an ophthalmic emergency, can cause corneal perforation and even endophthalmitis, thus leading to severe visual impairment. To achieve effective treatment of bacterial keratitis, good bioavailability of antimicrobial drugs on the ocular surface is desired. In this investigation, a layer-by-layer (LBL) self-assembly combined with the host-guest recognition was used to construct an antibacterial coating on the surface of corneal contact lens (CLs) to improve drug bioavailability and achieve successful treatment of bacterial keratitis. First, a radical copolymerization of acrylic acid (AA) and 1-adamantan-1-ylmethyl acrylate (AdA) was carried out to synthesize a polyanionic copolymer P(AA-co-AdA) (defined as PAcA). Then, PAcA copolymer combined with poly(ethyleneimine) (PEI) was used for a layer-by-layer (LBL) self-assembly to fabricate multilayer films on the surface of CLs. An antibacterial conjugate, β-cyclodextrin-levofloxacin (β-CD-LEV), was successfully synthesized and utilized to generate antibacterial coating through a host-guest interaction between AdA and β-CD-LEV. The antibacterial ability and treatment effect of bacterial keratitis was evaluated by in vitro assay and in vivo test in an animal model of staphylococcal keratitis, demonstrating that the antibacterial coating had good antibacterial and germicidal efficacy both in vivo and in vitro. We believe that this work will provide a promising strategy for the treatment of bacterial keratitis.


miMsg: a target enrichment algorithm for predicted miR-mRNA interactions based on relative ranking of matched expression data.
Algorithms predicting microRNA (miR)-mRNA interactions generate high numbers of possible interactions, many of which might be non-existent or irrelevant in a certain biological context. It is desirable to develop a transparent, user-friendly, unbiased tool to enrich miR-mRNA predictions. The miMsg algorithm uses matched miR/mRNA expression data to enrich miR-mRNA predictions. It grades interactions by the number, magnitude and significance of misplacements in the combined ranking profiles of miR/mRNA expression assessed over multiple biological samples. miMsg requires minimal user input and makes no statistical assumptions. It identified 921 out of 56 262 interactions as top scoring and significant in an actual germ cell cancer dataset. Twenty-eight miR-mRNA pairs were deemed of highest interest based on ranking by miMsg and supported by current knowledge about validated interactions and biological function. To conclude, miMsg is an effective algorithm to reduce a high number of predicted interactions to a small set of high confidence interactions for further study. Matlab source code and datasets available at www.martinrijlaarsdam.nl/mimsg . Supplementary data are available at Bioinformatics online.


Evaluation of hybrid rye on growth performance, carcass traits, and efficiency of net energy utilization in finishing steers.
Crossbred beef steers with a high percentage of Angus ancestry [n = 240, initial shrunk body weight (BW), 404 ± 18.5 kg] were used in a 117-d feedlot experiment to evaluate the effect of hybrid rye (Rye; KWS Cereals USA, LLC, Champaign, IL) as a replacement for dry-rolled corn (DRC) on growth performance, carcass traits, and comparative net energy (NE) value in diets fed to finishing steers. Rye from a single hybrid (KWS Bono) with an ergot alkaloid concentration of 392 ppb was processed with a roller mill to a processing index (PI) of 78.8 ± 2.29. Four treatments were used in a completely randomized design (n = 6 pens/treatment; 10 steers/pen), where DRC (PI = 86.9 ± 4.19) was replaced by varying proportions of Rye [DRC:Rye, dry matter (DM) basis (60:0), (40:20), (20:40), and (0:60)]. Liver abscess scores and carcass characteristics were collected at the abattoir. Carcass-adjusted performance was calculated from hot carcass weight (HCW)/0.625. Performance-adjusted NE was calculated using carcass-adjusted average daily gain (ADG), DM intake (DMI), and mean equivalent shrunk BW with the comparative NE values for rye calculated using the replacement technique. Data were analyzed using the GLIMMIX procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC) with pen as the experimental unit. Treatment effects were tested using linear and quadratic contrasts, as well as between diets with and without Rye. Replacing DRC with Rye linearly decreased (P ≤ 0.01) carcass-adjusted final BW, ADG, DMI, and gain:feed (G:F). Feeding rye linearly decreased HCW and longissimus muscle area (P ≤ 0.04). Distributions of liver scores and USDA grades for quality and yield were unaffected by treatment (P ≥ 0.09). Estimated replacement NE for maintenance (NEm) and gain (NEg) values for rye, when included at 60% of diet DM, were 1.90 and 1.25 Mcal/kg, respectively. Rye can be a suitable feed ingredient in finishing diets for feedlot steers. Estimated replacement values of Rye when fed at 60% of diet DM closely agreed with current tabular standards but, when included at 20% of diet DM, estimated NEm and NEg values of Rye were increased 9.5% and 12.8%, respectively. Net energy value of Rye for gain is approximately 84% compared to DRC; thus, the complete replacement of DRC with Rye depressed DMI, ADG, G:F, and carcass weight.


Behavior of hyaluronic acid from gingival epithelium and connective tissue on the analytical ultracentrifuge.
The molecular weights of hyaluronic acid (HA) isolated from separated specimens of human gingival epithelium and connective tissue as well as standard hyaluronic acid preparations have been estimated. The values were determined following substitution of sedimentation values into a previously determined empirical relationship between the reciprocal of the sedimentation coefficient at zero concentration (S-1)0 and molecular weights estimated by sedimentation-diffusion (MsD). The values of (S-1)0 for connective tissue and standard low-molecular weight HA preparations obtained by linear regression of all points indicated molecular weights (MsD) of 340,000 and 205,000 respectively. However, epithelial and standard high-molecular weight HA behaved differently during ultracentrifugation generating a curvilinear relationship between s-1 and concentration. Nevertheless linear extrapolation of a line of best fit of the very lowest concentrations (those which approached zero concentration) provided molecular weight estimates of 860,000 and 2,500,000 respectively. Moreover, similar treatment of s-1 values derived from the previously published data of Laurent, Ryan and Pietruszkiewicz has validated the use of linear regression of s-1 at the lower concentrations alone, to calculate high-molecular weight HA. The curvilinear relationship for s-1 throughout the whole concentration range (0.15-2.3 mg/ml) has been regarded only as a qualitative indication that the HA samples are of relatively high-molecular weight, while a straight line through such data points implies a qualitatively lower molecular weight for HA.


Studies on the nature and activation of O2(-)-forming NADPH oxidase of leukocytes. Identification of a phosphorylated component of the active enzyme.
Highly active superoxide (O2-)-forming NADPH oxidase was extracted from plasmamembranes of phorbol-12-myristate-13-acetate-activated pig neutrophils and was partially purified by gel filtration chromatography. Oxidase activity copurified with cytochrome b-245 in an aggregate containing phospholipids and was almost completely separated from FAD and NAD(P)H-cytochrome c reductase. A polypeptide with molecular weight of 31,500 strictly paralleled the purification of NADPH oxidase, suggesting that it is a major component of the enzyme. The enzyme complex was then dissociated by high detergent and salt concentration and cytochrome b-245 was isolated by a further gel filtration chromatography, with a 147 fold purification with respect to the initial preparation. The cytochrome b-245 showed a 31,500 molecular weight by SDS electrophoresis, indicating that it is actually the component previously identified in the partially purified enzyme. The 31,500 protein was phosphorylated in enzyme preparations from activated but not from resting neutrophils, suggesting that phosphorylation of cytochrome b-245 is involved in the activation mechanism of the O2(-) -forming enzyme responsible for the respiratory burst in phagocytes.


Breast cancer among women over 75 years: an important public health problem?
Women aged >75 years are not invited for mammographic screening; if diagnosed with breast cancer, due to their anticipated short-life expectancy, they are expected to die of other causes. To describe the breast cancer health problem in women aged >75 years, we estimated breast cancer incidence in this age group and the risk of breast cancer death in patients diagnosed after 75 years of age in Nijmegen, the Netherlands. Our findings demonstrate that in this age group, 3.3% of the women will be diagnosed with breast cancer, and that one in three of these incident cases die of this disease. These patients could have benefited from continued screening.


The Josephson effect between triplet superconductors through a finite ferromagnetic barrier.
Charge and spin transport in a junction involving two triplet superconductors and a ferromagnetic barrier are studied. We use Bogoliubov-de Gennes wavefunctions to construct the Green's function, from which we obtain the Josephson currents in terms of the Andreev reflection coefficients. We focus on the consequences of a finite barrier width for the occurrence of 0-π transitions and for the spin currents, and examine the appropriateness of the common δ-function approximation for the tunneling region.


A multicentre study of clinical features and HLA typing in Japanese patients with ankylosing spondylitis.
Due to the low prevalence of HLA-B27 and ankylosing spondylitis (AS) in Japan, rheumatologists have little experience with AS. We conducted a multicentre study to identify the characteristics and frequency of HLA-B types. We analysed epidemiological and clinical data, blood tests, spine radiographs, and HLA-B types in Japanese AS patients. We evaluated 111 AS patients, predominantly men (82.9%). The mean age, disease onset, diagnosis, and time from onset to diagnosis were 43.7, 24.2, 36.0, and 11.6 years, respectively. Inflammatory low back pain was found in 96 cases (86.5%); peripheral arthritis in 59 (53.2%), enthesitis in 35 (31.5%), and dactylitis in 6 (5.4%). Extra-articular symptoms included uveitis, psoriasis, and inflammatory bowel disease in 41 (36.9%), 1 (0.9%), and 5 (4.5%) cases, respectively. HLA-B27 was positive in 83 cases (74.8%; odds ratio, 1146.0); and HLA-B48 in 9 (8.1%; odds ratio, 3.0). HLA-B27-positive patients were younger at onset and had a shorter diagnostic delay. AS clinical symptoms were almost the same as other countries except for the low coexistence of psoriasis. HLA-B27 positivity in Japanese patients was 78%. HLA-B27-positive patients were younger and diagnosed earlier. In addition to HLA-B27, a relationship with HLA-B48 was suggested.


Classical Wigner theory of gas surface scattering.
The scattering of atoms from surfaces is studied within the classical Wigner formalism. A new analytical expression is derived for the angular distribution and its surface temperature dependence. The expression is valid in the limit of weak coupling between the vertical motion with respect to the surface and the horizontal motion of the atom along the periodic surface. The surface temperature dependence is obtained in the limit of weak coupling between the horizontal atomic motion and the surface phonons. The resulting expression, which takes into account the surface corrugation, leads to an almost symmetric double peaked angular distribution, with peaks at the rainbow angles. The analytic expression agrees with model numerical computations. It provides a good qualitative description for the experimentally measured angular distribution of Ne and Ar scattered from a Cu surface.


HAL fluorescence cystoscopy and TURB one year of Romanian experience.
Hexaminolevulinate blue light cystoscopy (HAL-BLC) represents an increasingly acknowledged method in bladder cancer diagnostic. We aimed to evaluate the importance of this procedure in cases of non-invasive bladder tumors (NIBT), to compare it with standard white light cystoscopy (WLC), and to establish the efficiency of blue light transurethral bladder resection (BL-TURB). Between December 2007 and December 2008, WLC and BLC were performed in 70 cases. WL-TURB was performed for all lesions visible in WL, and BL-TURB for those only detected in BL. Patients diagnosed with NIBT were followed-up after an average period of 5 months (between 18 and 22 weeks) by WLC and BLC. The control group included the same number of consecutive cases of NIBT, which underwent only WLC and WL-TURB, as well as the same follow-up protocol as the study group. WLC correctly identified 115 tumors, and BLC, 157 tumors. The detection rate was 68.8% for WLC, with a 9.4% rate of false-positive results, and 94% for BLC, with a 14.6% rate of false-positive results. The diagnostic accuracy in CIS lesions was 57.3% for WLC and 95% for BLC, while in pTa tumors; it was 68.8% for WLC and 94% for BLC. 62 cases of the study group diagnosed with NIBT emphasized a recurrence rate of 6.4% after 5 months. The control group described a recurrence rate of 24.2%. HAL fluorescence cystoscopy is a valuable diagnostic method for patients with NIBT, with considerably improved accuracy by comparison to WLC, and a significant impact upon the short-term recurrence rate.


Are pre-operative vocal fold checks necessary?
Over a 26-year period, from 1970 to 1995, 2250 pre-operative vocal fold checks were carried out by the ENT team at Manchester Royal Infirmary in patients undergoing thyroid, parathyroid and cardiothoracic surgery. A register was kept of all pre-operative fold checks. This register was retrospectively analysed and 23 vocal fold palsies were documented. In six of these patients the notes were either missing or incomplete. All of the remaining 17 patients had a history suggestive of vocal fold palsy. We suggest that in the absence of symptoms, either current or on history, a routine pre-operative vocal fold check is unnecessary.


Development and validation of a dried blood spot LC-MS/MS assay to quantify ranitidine in paediatric samples.
A novel approach has been developed to determine ranitidine in paediatric samples using dried blood spots (DBS) on Guthrie cards (Whatman 903). A selective and sensitive HPLC-MS/MS assay has been developed and validated using small volumes of blood (30 μl). A 6 mm disc was punched from each DBS and extracted with methanolic solution of the internal standard (IS) nizatidine. This was further subjected to solid phase extraction (SPE), followed by reversed phase HPLC separation, using a XBridge™ C18 column and mobile phase 10 mM ammonium acetate/methanol (98:2 v/v) with a flow rate of 0.3 mL/min. This was combined with multiple reaction monitoring (MRM) mass detection using electrospray ionisation (ESI). The calibration curve for ranitidine was found linear over the range 10-500 ng/mL (r=0.996). The limit of quantification (LOQ) of the method was validated at 10 ng/mL. Accuracy and precision values for within and between days were <20% at the LOQ and <15% at all other concentrations. The validated DBS method was successfully applied to a clinical study employing 81 samples from 36 paediatric patients.


Molecular Dynamics Simulations for Resolving Scaling Laws of Polyethylene Melts.
Long-timescale molecular dynamics simulations were performed to estimate the actual physical nature of a united-atom model of polyethylene (PE). Several scaling laws for representative polymer properties are compared to theoretical predictions. Internal structure results indicate a clear departure from theoretical predictions that assume ideal chain statics. Chain motion deviates from predictions that assume ideal motion of short chains. With regard to linear viscoelasticity, the presence or absence of entanglements strongly affects the duration of the theoretical behavior. Overall, the results indicate that Gaussian statics and dynamics are not necessarily established for real atomistic models of PE. Moreover, the actual physical nature should be carefully considered when using atomistic models for applications that expect typical polymer behaviors.


Messenger ribonucleic acid encoding interferon-inducible guanylate binding protein 1 is induced in human endometrium within the putative window of implantation.
The putative window of embryo implantation in the human opens between days 19--24 of the menstrual cycle. During this period, the endometrium undergoes distinctive structural and functional changes orchestrated by steroid hormones, growth factors, and cytokines to attain a receptive phase in which it acquires the ability to implant the developing embryo. A major challenge in the study of human reproduction is to identify the molecular signals that participate in the establishment of this critical receptive phase in the context of the natural cycle. Toward this goal, we analyzed human endometrial biopsies at various days of the menstrual cycle by employing messenger RNA (mRNA) differential display technique. We isolated several complementary DNAs representing genes that are either up- or down-regulated within the putative window of implantation. We identified one of these genes as that encoding interferon (IFN)-inducible guanylate-binding protein 1 (or GBP1), which possesses GTPase activity. Analysis of endometrial biopsies by Northern blotting and RT-PCR demonstrated that GBP1 mRNA is specifically induced at the midsecretory phase of the menstrual cycle. In situ hybridization analysis revealed that GBP1 mRNA expression is localized in the glandular epithelial cells as well as in the stroma in the immediate vicinity of the glands. We observed that treatment of human endometrial adenocarcinoma cell, Ishikawa, with IFN-gamma or IFN-alpha markedly induced the expression of GBP1 mRNA. IFN-gamma was, however, a more potent inducer of GBP1 than IFN-alpha. Consistent with this finding, the temporal profile of GBP1 expression during the menstrual cycle resembled that of IFN-gamma mRNA more closely than that of IFN-alpha, predicting a regulatory role of IFN-gamma in GBP1 expression in midsecretory human endometrium. Although the precise function of GBP1 in the receptive human uterus remains unclear, its unique expression overlapping the putative window of implantation suggests that it might serve as a useful marker of uterine receptivity in the human.


Differential Activation Patterns in the Same Brain Region Led to Opposite Emotional States.
In human studies, how averaged activation in a brain region relates to human behavior has been extensively investigated. This approach has led to the finding that positive and negative facial preferences are represented by different brain regions. However, using a functional magnetic resonance imaging (fMRI) decoded neurofeedback (DecNef) method, we found that different patterns of neural activations within the cingulate cortex (CC) play roles in representing opposite directions of facial preference. In the present study, while neutrally preferred faces were presented, multi-voxel activation patterns in the CC that corresponded to higher (or lower) preference were repeatedly induced by fMRI DecNef. As a result, previously neutrally preferred faces became more (or less) preferred. We conclude that a different activation pattern in the CC, rather than averaged activation in a different area, represents and suffices to determine positive or negative facial preference. This new approach may reveal the importance of an activation pattern within a brain region in many cognitive functions.


WRKY70 prevents axenic activation of plant immunity by direct repression of SARD1.
SARD1 is an activator of plant immunity that promotes production of the hormone salicylic acid (SA) and activation of defense gene expression. SARD1 itself is strongly inducible by infection. Here, we investigated the transcriptional control of SARD1. We used yeast one-hybrid assays to identify WRKY70. The WRKY70 binding site was defined using electrophoretic mobility shift assays, and its importance was investigated using an Arabidopsis thaliana protoplast system. The effect of wrky70 mutations was studied by measurements of pathogen growth, SA concentrations, and gene expression by RNA-seq. WRKY70 binds to a GACTTTT motif in the SARD1 promoter in yeast and Arabidopsis protoplasts. Plants with wrky70 mutations have elevated expression of SARD1 in the absence of pathogens, but not when infected. Expression profiling revealed that WRKY70 represses many pathogen-inducible genes in the absence of pathogens, yet is required for activation of many other pathogen-inducible genes in infected plants. The GACTTTT motif is enriched in the promoters of both these gene sets, and conserved in SARD1 orthologs within the Brassicaceae. WRKY70 represses SARD1 by binding the motif GACTTTT in the absence of pathogens. Conservation of the WRKY70 binding among the Brassicaceae suggests that WRKY70 repression of SARD1 is important for fitness.


DOCA-salt hypertensive rat hearts exhibit altered expression of G-proteins.
We have recently demonstrated an alteration in the levels of G-proteins and their correlation with adenylyl cyclase in spontaneously hypertensive rats (SHR). In the present studies we examined if the other models of hypertensive rats, such as DOCA-salt hypertensive rats (HR), also exhibit the similar alterations in G-protein and in adenylyl cyclase activity. We have determined the adenylyl cyclase activity stimulated and inhibited by hormones, as well as the levels of G-proteins using specific antibodies and cDNA probes in the hearts from DOCA-salt HR and their sham-operated controls after 2 and 4 weeks of treatment. Adenylyl cyclase activity stimulated by GTP gamma S, isoproterenol, and glucagon was significantly decreased in heart sarcolemma from DOCA-salt HR as compared to their controls after 2 and 4 weeks of treatment. In addition, the inhibitory hormones inhibited the enzyme activity to a greater extent in hypertensive rats than controls. Furthermore, the levels of Gi alpha-2 and Gi alpha-2 mRNA, as determined by immunoblotting and Northern blotting techniques, respectively, were higher in hearts from DOCA-salt HR. However, the levels of G8 alpha 45 were decreased in these rats. These results indicate that, similar to SHR, the hearts from DOCA-salt HR exhibit the increased expression of Gi, however unlike SHR, the expression of G8 was decreased. It is suggested that the altered expression of G-proteins may partly be responsible for the decreased responsiveness of adenylyl cyclase to hormone stimulation and increased responsiveness to hormone inhibition in DOCA-salt hypertension.


[Current status of radical laparoscopic prostatectomy].
Laparoscopic surgery can be said to have come of age when it was first indicated for cancer conditions. Advances in this field are largely due to the French school, which has made it a standard practise in prostate cancer. It complies with the principles required for cancer as well as conventional surgery, but it remains to be verified whether its long-term results, both from tumoral and functional perspectives, are not only similar to those of classical surgery, but even better. In fact, increasing numbers of clinical groups are incorporating this technique in their daily work.


Percutaneous dilatational tracheostomy is as safe as open tracheostomy.
Although percutaneous dilatational tracheostomy (PDT) has been advocated as an alternative to open tracheostomy (OT) its relative safety has been questioned repeatedly. This study prospectively compared the safety and complications of PDT and OT. Ninety-four patients underwent PDT and 252 patients underwent OT at this institution from December 1998 through April 2000 with the choice of procedure left to the operator. OT was performed in the operating room whereas PDT was performed in intensive care units (ICUs). PDT was performed by surgeons and medical intensivists under a strict institutional policy and procedure governing patient selection and conduct of the procedure. Complications were defined as bleeding, loss of airway, hypotension, hypoxia, tracheostomy tube malposition, subcutaneous emphysema, infection, and conversion of PDT to OT. All patients survived the operation. PDT and OT had similar complication rates: 2.1 per cent for PDT versus 2.8 per cent for OT (P = not significant). Postoperative bleeding, which was the most frequent complication, occurred in one PDT patient and four OT patients. One PDT patient required conversion to OT as a result of extensive tracheal fibrosis. Subcutaneous emphysema, soft-tissue infection, and a malpositioned tracheostomy tube were the remaining complications in the OT patients. We conclude that the complication rates of PDT and OT are comparable. The choice of PDT or OT should be dictated by the surgeon's training and experience, the patient's condition, neck anatomy, and stability for transfer to the operating room.


[Study on the law of circulation of meridians].
To study the basic law of circulation of channels and collaterals. Inherit and develop ripe experiences of predecessors based on The Yellow Emperor's Internal Classic and other classic medical books. Circulation of channels and collaterals has the eight laws, including naming law, distribution law, converging law, exterior-interior association law, beginning-ending running law, meridian-qi bidirectional circulation law, zang- and fu-organ pathway liaison law, and liaison law of connecting with trunks and sense organs.


Lateral Collateral Ligament and Biceps Femoris Tendon Fixation with a Suture Anchor to the Tibial Metaphysis after Proximal Fibula En Bloc Resections Preserve Lateral Knee Stability.
This study aims to analyze the functional outcomes and lateral knee stability of patients who underwent lateral collateral ligament (LCL) and biceps femoris tendon reconstruction with suture anchors after proximal fibula en bloc resection for bone tumors. Patients who underwent proximal fibular en bloc resection between 2007 and 2018 were retrospectively viewed. Patients were invited to visit the clinic, and their functional scores were evaluated using the Musculoskeletal Tumor Society Scoring (MSTS) system. Lateral knee stability was evaluated by varus stress radiographs obtained at 20 degrees of flexion, and the range of motion (ROM) of the bilateral knee was assessed. Side-to-side differences were graded according to the International Knee Documentation Committee (IKDC) criteria and compared between types I and II resections. A total of 17 patients (4 males and 13 females) with a mean age of 31.1 ± 17.1 (range: 13-65) years at the time of surgery were available for radiological and clinical examination at a mean follow-up of 68.6 ± 36.4 (range: 22-124) months after surgery. In terms of ROM measurements, IKDC grades and side-to-side differences in both flexion and extension were not significantly different between the groups. On varus stress radiographs, lateral knee gapping was measured to be 0.93 ± 0.91 mm in type-I resections and 1.83 ± 0.45 mm in type-II resections, and statistically significant differences were detected among the groups (p = 0.039). When the values were graded according to IKDC criteria, none of the knees were classified as abnormal, and no difference was observed between the groups. Mean MSTS score of patients with type-I resections was significantly higher than those of patients with type-II resections (92.7 vs. 84.4%, p = 0.021). In the subscale analysis, a significant difference was observed in the support scores (type I = 94.5%, type II = 70%; p = 0.001). The reattachment of LCL and biceps femoris tendon to the tibial metaphysis with a suture anchor is a simple and effective method to prevent lateral knee instability after proximal fibula resections.


Human erythrocyte nucleoside transporter ENT1 functions at ice-cold temperatures.
The functionality of human erythrocyte nucleoside transporter ENT1 was examined at ice-cold temperatures (ICT; measured temperature, 0.5-0.7 degrees C) using rightside-out membrane vesicles (ROVs). The uptake of uridine, an ENT1 substrate, showed saturation kinetics and was inhibited by S-(4-nitrobenzyl)-6-thioinosine (NBMPR), a specific ENT1 inhibitor, at both 23 degrees C and ICT. [3H]Uridine uptake was markedly trans-stimulated by preloading ROVs with unlabeled uridine or ribavirin, another ENT1 substrate, and the overshoot phenomenon was observed at ICT. Similarly, [3H]ribavirin uptake was markedly trans-stimulated by unlabeled ribavirin or uridine at ICT. The trans-stimulated uptake of [3H]uridine at ICT was inhibited by ENT1 inhibitors/substrates such as NBMPR, dipyridamole, adenosine, and ribavirin in a concentration-dependent manner. The inhibition of [3H]uridine uptake by NBMPR and dipyridamole at ICT was also observed in intact red blood cells. Like uridine uptake, [3H]D-glucose uptake by ROVs, which is mediated by facilitative glucose transporter GLUT1, was trans-stimulated by unlabeled D-glucose at ICT, and the overshoot phenomenon was observed. In contrast, the ability of ATP-dependent transport of 5-(and-6)-carboxy-2',7'-dichlorofluorescein via multidrug resistance-associated protein 5 in inside-out membrane vesicles disappeared at ICT. These results clearly indicate that human erythrocyte transporters such as ENT1 function even at very low temperatures near 0 degrees C. The significance of these findings in transporter research is discussed.


Involvement of hydroxyl radicals in neurogenic airway plasma exudation and bronchoconstriction in guinea-pigs in vivo.
1. Cigarette smoke induces plasma exudation in the airways of rodents by activation of capsaicin-sensitive 'sensory-efferent' nerves. The response is mediated predominantly by substance P (SP) and the magnitude of exudation is regulated by neutral endopeptidase (NEP). The component(s) of the smoke responsible for the activation of the nerves may be reactive oxygen radicals. We investigated the effect of the hydroxyl radical scavenger dimethylthiourea (DMTU), a regulator of superoxide anion, superoxide dismutase (SOD), and a regulator of hydrogen peroxide, catalase, on plasma exudation (measured using Evans blue dye) induced by cigarette smoke in guinea-pig main bronchi in vivo. The effect of DMTU on plasma exudation and non-cholinergic bronchoconstriction (measured as pulmonary insufflation pressure, PIP) induced by electrical stimulation of the vagus nerves was also assessed. Interaction between hydroxyl radicals and NEP was assessed with the NEP inhibitor phosphoramidon. 2. In each of the experiments, cigarette smoke increased plasma exudation by approximately 200% above air-exposed controls. Acute administration of DMTU (1.5 g kg-1, i.v. for 20 min) significantly reduced cigarette smoke-induced plasma exudation by 69%. In contrast, neither SOD (240,000 u kg-1, i.v.) nor catalase (400,000 u kg-1, i.v.) significantly affected the exudative response. 3. Chronic pretreatment with DMTU (1.25 g kg-1 over 4 days) significantly reduced bronchial plasma exudation induced by cigarette smoke by 72%. Phosphoramidon (1.5 mg kg-1, i.v.) completely reversed the inhibition by DMTU of cigarette smoke-induced plasma exudation. 4. Vagal stimulation increased plasma exudation by approximately 200% and PIP by approximately 250%. Acute treatment with DMTU had no significant inhibitory effect on these responses, whereas chronic pretreatment inhibited them by approximately 80%. Phosphoramidon reversed the inhibition by chronic DMTU. 5. SP (1 nmol kg-1) increased plasma exudation by approximately 250%, a response which was not inhibited by either acute or chronic DMTU. 6. We conclude that hydroxyl radicals, rather than superoxide anion or hydrogen peroxide, are involved in the induction of neurogenic plasma exudation and bronchoconstriction induced by cigarette smoke or by electrical stimulation of the vagus nerves. These radicals also affect the activity of NEP. Acute DMTU may affect directly the neural actions of hydroxyl radicals contained in the cigarette smoke. Chronic pretreatment with DMTU may inhibit the neurogenic airway responses by effects on tachykinin biosynthesis and/or axonal transport.


Study of the interaction between hair protein and organic acid that improves hair-set durability by near-infrared spectroscopy.
In this study, hydrogen bonds around hair proteins were analyzed by near-infrared spectroscopy to reveal the mechanism of improving hair-set durability by treatment with a specific organic acid. The improvement of set durability was confirmed by measurement on single hair fibers, suggesting that improvement is not because of the surface adhesion increase but because of the internal changes in the hair. Through analysis by two-dimensional near-infrared correlation spectroscopy, it was found that a combination band of stretching NH and amide II is deconvoluted into three bands interacting with different hydrogen bonds. From the assignment of the three bands, the behavior of the organic acid in the hair was clarified as follows: it absorbs at the site where water originally binds, even in extremely dry conditions, prevents water penetration, and makes strong and stable hydrogen bonds with hair proteins. The formation of such strong and stable hydrogen bonds suppresses the exchange of hydrogen bonds that is the cause of the breakage of set durability.


Sequence diversity of Jeryl Lynn strain of mumps virus: quantitative mutant analysis for vaccine quality control.
The Jeryl Lynn strain of mumps vaccine live (MVL) was developed in 1966 by Merck Co. and has been widely used in the U.S. and other countries since the early 1970s. Partial sequencing has recently shown that the vaccine contains a mixture of two substrains with substantially different nucleotide sequences. We have determined the complete genomic sequences of both substrains and identified 414 nucleotide differences (2.69%), leading to 87 amino acid substitutions (1.67%). We used this information to develop methods for quantification of the substrain components in vaccine samples based on PCR and restriction enzyme cleavage and oligonucleotide microarray hybridization and monitored their dynamics in viral populations propagated in different conditions. Passaging Jeryl Lynn strain in Vero or CEF cell cultures resulted in rapid selection of the major component JL1, while growth in embryonated chicken eggs (ECE) favored accumulation of the minor component JL2. Based on the findings presented here, it is proposed that the substrain composition of Jeryl Lynn vaccine can be monitored as a part of its quality control to ensure consistency of the vaccine.


Investigation of the C-3-epi-25(OH)D3 of 25-hydroxyvitamin D3 in urban schoolchildren.
The physiological relevance C-3 epimer of 25-hydroxyvitamin D (3-epi-25(OH)D) is not well understood among youth. The objective of this study was to assess whether demographic/physiologic characteristics were associated with 3-epi-25(OH)D3 concentrations in youth. Associations between 3-epi-25(OH)D3 and demographics and between 3-epi-25(OH)D3, total 25-hydroxyvitamin (25(OH)D) (25(OH)D2 + 25(OH)D3), total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides were examined in racially/ethnically diverse schoolchildren (n = 682; age, 8-15 years) at Boston-area urban schools. Approximately 50% of participants had detectable 3-epi-25(OH)D3 (range 0.95-3.95 ng/mL). The percentage of 3-epi-25(OH)D3 of total 25(OH)D ranged from 2.5% to 17.0% (median 5.5%). Males were 38% more likely than females to have detectable 3-epi-25(OH)D3 concentrations. Both Asian and black race/ethnicity were associated with lower odds of having detectable 3-epi-25(OH)D3 compared with non-Hispanic white children (Asian vs. white, odds ratio (OR) 0.28, 95% confidence interval (CI) 0.14-0.53; black vs. white, OR 0.38, 95%CI 0.23-0.63, p < 0.001). Having an adequate (20-29 ng/mL) or optimal (>30 ng/mL) 25(OH)D concentration was associated with higher odds of having detectable 3-epi-25(OH)D3 than having an inadequate (<20 ng/mL) concentration (OR 4.78, 95%CI 3.23-6.94 or OR 14.10, 95%CI 7.10-28.0, respectively). There was no association between 3-epi-25(OH)D3 and blood lipids. However, when considering 3-epi-25(OH)D3 as a percentage of total 25(OH)D, total cholesterol was lower in children with percent 3-epi-25(OH)D3 above the median (mean difference -7.1 mg/dL, p = 0.01). In conclusion, among schoolchildren, sex, race/ethnicity, and total serum 25(OH)D concentration is differentially associated with 3-epi-25(OH)D. The physiological relevance of 3-epi-25(OH)D3 may be related to the 3-epi-25(OH)D3 as a percentage of total 25(OH)D and should be considered in future investigations.


Alcohol and inflammation: a possible mechanism for protection against ischemic heart disease.
Moderate alcohol consumption has been associated with a decreased risk for ischemic heart disease. Increases in high-density lipoprotein-cholesterol may explain approximately 50% of this apparent benefit. Another potential mechanism may involve the effects of alcohol on systemic inflammation. Accordingly, the purpose of this article is to provide a narrative review of the literature concerning alcohol and inflammation. Relevant articles were identified by searching MEDLINE from 1966 to 2001. Search terms included alcohol or ethanol combined with inflammation, interleukin-1, interleukin-6, or tumor necrosis factor alpha. Important references from these articles were also identified and reviewed. Considerable evidence exists from animal and human studies for potentially anti-inflammatory effects of alcohol consumption. Further research is needed to determine the effects of various patterns and levels of alcohol consumption on systemic markers of inflammation in humans.


Follow-up study of patients treated with monoclonal antibody-drug conjugate: report of 77 cases with colorectal cancer.
A total of 77 patients with advanced colorectal cancer, including postoperative patients with liver, lung and peritoneal metastases, were treated with single or multiple injections of monoclonal antibody A7-neocarzinostatin (A7-NCS). A follow-up study of the patients treated with A7-NCS was done and the clinical outcome was compared with that of patients given other chemotherapies. In the postoperative patients with liver metastasis, the A7-NCS treatment prolonged survival time when compared with systemic administration of anticancer drugs, while it showed a similar survival time to chemoembolization using multiple anticancer agents suspended in a lipid contrast medium. Among the patients who underwent surgical resection of primary cancer, with or without liver metastasis, there was no difference in overall 5-year survival rate between the group treated with A7-NCS and the group treated with the other chemotherapies. However, the survival time of the patients treated with A7-NCS was longer than that of the patients treated with the other chemotherapies. In addition, the patients given a higher dose of A7-NCS had a longer survival time than the patients given a lower dose of A7-NCS. Human anti-mouse antibody was detected in all the A7-NCS-treated patients examined. There were no serious side effects in any of the patients given A7-NCS. Thus, this study indicates that the A7-NCS treatment is safe and useful for colorectal cancer patients, though some problems remain, such as optimization of injection dose, route, interval, etc., and overcoming human anti-mouse antibody development.


[Diabetic examination in general practice. Screening of a practice population and a study of occurrence].
In a general practice, a screening examination for diabetes mellitus by measuring blood glucose in capillary blood was performed. A total of 2,843 persons aged 20-80 years were invited, 1,973 (69.4%) participated. At screening, 66 possible cases (BG greater than or equal to 8 mmol/l) were found. Among these, 16 diabetics were found, nine of whom had predisposing conditions. The prevalence of diabetes mellitus in the examined and known population of the practice was found to be 2.5% and calculated to be 2.2% in the total population of the practice. It was demonstrated that an investigation of this kind can be carried out but felt laborious and interfered with the daily work. Nevertheless, hitherto unknown diabetics were detected and instructed. A method of finding diabetics in general practice is thus described. The method is probably effective, as the prevalence of diabetes mellitus reached the expected, when the known and newly found diabetics were added.


Chronic Kidney Disease Education Class Improves Rates of Early Access Creation and Peritoneal Dialysis Enrollment.
Background Most patients with end-stage kidney disease begin hemodialysis (HD) in an unplanned fashion at a late stage, necessitating the commencement of HD with a temporary venous catheter, the least favorable option. Alternative modalities of kidney replacement therapy (KRT), peritoneal dialysis (PD), and preemptive transplant offer similar or better outcomes than HD at a lower overall cost, and yet they remain underutilized in Saudi Arabia. Early education may help prepare patients with advanced chronic kidney disease (CKD IV and V) to accept their disease and choose a KRT modality that minimizes complications and matches their lifestyle. The aim of the study is to assess the impact of a pilot educational class on therapy choices and outcomes. Methodology In a cross-sectional study, we conducted phone interviews and reviewed medical records of 81 attendees of the multidisciplinary monthly educational class about KRT that was held at the King Abdulaziz Medical City (KAMC) from January 2017 to October 2021. The interview was conducted at least one year after the participants attended the class. The study proposal, consent, and questionnaire were approved by the King Abdulaziz International Medical Research Center. Patient data was retrieved from KAMC electronic medical record system. Results Volunteer participation in the survey was high (62/81). For the respondents, a preemptive kidney transplant was the most preferred (48/62, 77%) option for KRT. Among the preferred fallback options, HD was the most frequently chosen (29/62, 47%) compared to PD (26/62, 41.9%). At the time of the interview, a great majority of the patients (54/62, 87%) was already on KRT, including about half (26/54, 48%) on HD via a catheter, and the rest about equally divided between those on HD via an arteriovenous (AF) fistula (13/54, 24%) and those on PD (15/54, 28%). Thus, half of the respondents on KRT (28/54, 51%) avoided urgent HD catheter commencement. However, because of an unfortunate shortage of donors, only a small minority (2/62, 3%) of patients received preemptive transplantation. Conclusion The KAMC CKD education class helped boost the fraction of patients, significantly above the national average, who accepted the diagnosis of kidney failure and pursued preemptive native HD access or enrolled in PD.


Evaluation of the aetiological spectrum of obstructive jaundice.
Jaundice is a common problem in medical and surgical gastroenterological practice. The surgical jaundice can be caused by the obstruction of the bile duct as with gall stones, strictures, malignancy, such as cholangiocarcinoma (in which the jaundice is persistent and progressive), periampullary carcinoma, carcinoma gall bladder and carcinoma head of pancreas. The objective of this descriptive study was to evaluate the Etiological spectrum of obstructive jaundice. A prospective, descriptive study was carried out at Surgical Unit-II Holy family Hospital, Rawalpindi, from mid of May 2006 till March 2007. Sixty patients, who presented in the surgical OPD of Holy family Hospital, were included in the study. Thorough history and physical examination was followed by biochemical tests and various investigations like USG abdomen, ERCP, CT-Scan, & MRCP and histopathology. The data was analyzed using SPSS ver 14.0. Of the 60 patients; 40 (66.66%) were male and 20 (33.33%) were female, their mean age being 49.50 years. Malignant obstructive jaundice was seen in 34 (56.66%) patients while 26 (43.33%) had benign etiology. Amongst the commonest symptom; clay coloured stools (75%) was more frequent in patients with malignant disease whereas abdominal pain (51.66%) was most common in benign conditions. Commonest malignancy was Carcinoma (Ca) of the head of pancreas 18/60 (30%) followed by Ca gall bladder 8/60 (13.33%), cholangiocarcinoma 7/60 (11.66%), and periampullary carcinoma 1/60 (1.66%). Choledocholithiasis 21/60 (35%) was the commonest benign cause followed by stricture of common bile duct 3/60 (5%) and acute pancreatitis 2/60 (3.33%). Obstructive jaundice is common amongst females and the cause is mostly malignant. Ca head of pancreas is the commonest malignancy while Choledocholithiasis is the commonest benign cause. USG, ERCP and CT-Scan are important diagnostic modalities for evaluation of patient with obstructive jaundice with ERCP having the additional advantage of being therapeutic as well.


Cattle infected with bovine leukaemia virus may not only develop persistent B-cell lymphocytosis but also persistent B-cell lymphopenia.
We investigated the distribution of B and T cells in the peripheral blood of haematologically inconspicuous (non-persistent lymphocytotic, PL-) cattle infected with the bovine leukaemia virus (BLV). Flow cytometric data were obtained from six PL- cattle and compared with six age-matched animals with persistent lymphocytosis (PL+) and five non-infected healthy controls (BLV-). In the PL- group, the percentage and number of surface immunoglobulin-positive (sIg+) B cells were significantly reduced. Whereas in BLV-cattle, about 40% of the peripheral blood lymphocytes (PBL) were sIg + and 24% were sIgM + B cells. In the PL- group, less than 20% of the PBL were sIg+ and sIgM+ B cells. Only 5% of the PBL co-expressed sIgM+ and CD5+ versus 16% in BLV-. This decrease was persistent over 3 years and predominantly affected: (i) B cells that did not express sIgM; (ii) sIgM + B cells co-expressing CD5 and CD11b; and (iii) equally both lambda- and K-type light chain B-cell subpopulations. In contrast, the number of all circulating lymphocytes, CD5- and CD11b- sIgM+ B cells and CD2+ T cells did not differ. In PL+ animals, about 75% of the PBL were sIgM+ CD5+ B cells. These cells were of polyclonal origin, as light chains of the lambda- and K-type were expressed in a ratio of 4:1 (57.7% of PBL lambda+, 14% kappa+) as in BLV- animals (33.6% of PBL lambda+, 8.7% kappa+). In PL+ cattle the absolute number of B-cells and, therefore, their relative percentage is significantly increased. For this reason, even in case of absolutely increased T-cell numbers, the relative percentage of T-cells could be lower than in normal controls. The cause for the observed B cell decrease in PL- cattle is unknown, but it can be assumed that cytotoxic T cells are involved in this B-cell lymphopenia.