A Glycine max homolog of NON-RACE SPECIFIC DISEASE RESISTANCE 1 (NDR1) alters defense gene expression while functioning during a resistance response to different root pathogens in different genetic backgrounds.
A Glycine max homolog of the Arabidopsis thaliana NON-RACE SPECIFIC DISEASE RESISTANCE 1 (NDR1) coiled-coil nucleotide binding leucine rich repeat (CC-NB-LRR) defense signaling gene (Gm-NDR1-1) is expressed in root cells undergoing a defense response to the root pathogenic nematode, Heterodera glycines. Gm-NDR1-1 overexpression in the H. glycines-susceptible genotype G. max[Williams 82/PI 518671] impairs parasitism. In contrast, Gm-NDR1-1 RNA interference (RNAi) in the H. glycines-resistant genotype G. max[Peking/PI 548402] facilitates parasitism. The broad effectiveness of Gm-NDR1-1 in impairing parasitism has then been examined by engineering its heterologous expression in Gossypium hirsutum which is susceptible to the root pathogenic nematode Meloidogyne incognita. The heterologous expression of Gm-NDR1-1 in G. hirsutum effectively impairs M. incognita parasitism, reducing gall, egg mass, egg and juvenile numbers. In contrast to our prior experiments examining the effectiveness of the heterologous expression of a G. max homolog of the A. thaliana salicyclic acid signaling (SA) gene NONEXPRESSOR OF PR1 (Gm-NPR1-2), no cumulative negative effect on M. incognita parasitism has been observed in G. hirsutum expressing Gm-NDR1-1. The results indicate a common genetic basis exists for plant resistance to parasitic nematodes that involves Gm-NDR1. However, the Gm-NDR1-1 functions in ways that are measurably dissimilar to Gm-NPR1-2. Notably, Gm-NDR1-1 overexpression leads to increased relative transcript levels of its homologs of A. thaliana genes functioning in SA signaling, including NPR1-2, TGA2-1 and LESION SIMULATING DISEASE1 (LSD1-2) that is lost in Gm-NDR1-1 RNAi lines. Similar observations have been made regarding the expression of other defense genes.


Effects of postoperative medical treatment and expectant treatment on dysmenorrhea after conservative laparoscopic surgery for deep-infiltrating endometriosis accompanied by dysmenorrhea.
To compare the efficacy of postoperative adjuvant treatment (gonadotropin-releasing hormone agonists [GnRHas] and oral contraceptives [OCs]) and expectant treatment in preventing recurrent dysmenorrhea following conservative laparoscopic surgery for deep infiltrating endometriosis (DIE) with dysmenorrhea. A prospective cohort study was conducted in Shanghai, China. In total, 147 patients with dysmenorrhea who underwent conservative laparoscopic surgery for DIE were enrolled. Following surgery, patients received either postoperative adjuvant therapy (GnRHa or OCs) for 6 months or expectant treatment according to a shared medical decision-making approach. The primary outcome was the postoperative recurrence of dysmenorrhea. The secondary outcomes included reproductive outcomes and drug-induced side effects. The generalized estimating equation analysis illustrated that the visual analog scale for dysmenorrhea was significantly higher in the adjuvant treatment group than in the expectant treatment group. Kaplan-Meier analysis and the log-rank test demonstrated that the cumulative recurrence rate was higher in the expectant treatment group than in the adjuvant treatment group, but no difference was noted between the two hormonal treatments. Similar cumulative 24-month clinical pregnancy rates were observed among the three groups. Compared with expectant management, postoperative medical treatment more effectively relieved symptoms and prevented the recurrence of dysmenorrhea.


Synthesis and Substrate Evaluation of (E)-5-[(3-Selenophene-2-Carboxamido)Prop-1-en-1-yl]-Uridine-5'-O-Triphosphate for RNA Polymerase.
Design, synthesis and T7 RNA polymerase substrate evaluation of (E)-5-[(3-selenophene-2-carboxamido)prop-1-en-1-yl]-uridine-5'-O-triphosphate is reported. The title compound is shown to be a good substrate for RNA polymerase by RNA labeling through in vitro transcription. pTRI-plasmid DNA with β-actin gene sequence (∼300 base pairs) with T7 promoter was used as a template for the in vitro transcription. Transcribed product is characterized for incorporation by gel assay and for integrity, full length and size by bioanalyzer. The title compound will be very useful in biophysical techniques to obtain information on dynamics and recognition properties in real time as well as 3D structure of nucleic acids.


Incidence and characterization of acute kidney injury after acetaminophen overdose.
Acute kidney injury (AKI) occurs in 2-10% of patients with acetaminophen (APAP) overdose. Elevation in creatinine (SCr) typically occurs 2 to 5 days after ingestion, with a mean peak on day 7, and normalization over a month. However, it remains unclear whether renal impairment occurs without hepatotoxicity. We hypothesized that APAP-associated acute renal failure occurs in patients with and without severe liver dysfunction after APAP overdose. We retrospectively evaluated all patients admitted to the Medical Intensive Care Unit at a tertiary hospital and received acetylcysteine between June 2009 and December 2014. Of the 303 patients meeting these criteria, 139 of these patients received acetylcysteine for APAP overdose. Of these patients, 138 had Model for End-Stage Liver Disease (MELD) Scores on Day 1 of admission. Using a modified MELD (m-MELD) score, only containing total bilirubin and international normalized ratio not the SCr, the median m-MELD score was calculated. Patients with m-MELD scores below the median were compared to those with scores above the median (low m-MELD score <2.9 or high m-MELD score >2.9). Baseline demographics were similar in the two groups with the exception of more hypertension in the low m-MELD group (24 vs 7%; P= .02). Time to admission was shorter in the low m-MELD group (7.9 ± 9.3 vs. 25.7 ± 29.2 hours; P= .001). The mean admission APAP level was 96.9 (±119) μg/mL in the low compared to 52.3 (±85.3) μg/mL in the high m-MELD group (P= .012). Day one SCr (1.2 ± 0.9 vs 2.7 ± 2.2 mg/dL; P< .0001) and change from baseline to highest SCr (0.2 ± 0.3 vs. 2.7 ± 3.3 mg/dL; P< .0001) were both lower in the low m-MELD group compared to the high m-MELD group. In addition, renal failure resolved upon discharge in all 2 patients (3%) with AKI in the low m-MELD group as compared to only 19 patients (44%) in the high m-MELD group. Mean day one SCr, maximum change in SCr, and lack of renal failure resolution were higher in patients with higher m-MELD scores. However, patients with low m-MELD scores presented much earlier than patients with high m-MELD scores and 26% developed AKI.


Post-radiation glioma in a child. Case report and review of the literature.
A case of primary brain neoplasm is reported that developed 11 years after prophylactic brain irradiation and intrathecal methotrexate (MTX) given for acute lymphocytic leukemia (ALL) at the age of 3 years. Cases of supposed postirradiation gliomas in children seem to have increased in recent years, particularly following apparently successful treatment for ALL or medulloblastoma. This increase might reflect the increased number of long-surviving cancer children and thus suggest that brain irradiation may be a potential risk factor in this population. After reviewing some general data on neurooncogenesis, two points are stressed: (1) the child's brain is particularly susceptible to the mutagenic effects of ionizing radiation owing to physiologically active postnatal glial cell proliferation, which takes place in the cerebral hemispheres, especially during myelinogenesis; (2) MTX could promote glial neoplasm by enhancing reparative glial cell proliferation. Prospective studies seem necessary to show whether or not prophylactic brain radiation in ALL plays an oncogenetic role. Nevertheless, besides recurrent leukemia, leukoencephalopathy, and radionecrosis, the remote possibility of radiation-induced glial neoplasms should always be considered in cases of children treated for leukemia who present with neurological complications.


Optimization of cryopreservation procedures for rat and human hepatocytes.
1. Rat hepatocytes were cryopreserved using a number of procedures and the viability, attachment, and metabolic activity of the cryopreserved cells were compared to freshly isolated hepatocytes. Several cryopreservation agents (dimethylsulphoxide [DMSO], glycerol, polyvinylpyrrolidone [PVP], dextrans), and combinations of these agents, were examined. Other variables tested included the freezing rate, thawing rate, and the concentration of serum in the freezing medium. 2. Recovery of viable attached cells was optimal using DMSO at concentrations of 10% or higher, a slow stepwise cooling procedure, and a quick thaw. The concentration of serum in the freezing medium (0% to 90%) did not affect cryopreservation results. Using this procedure the recovery of viable hepatocytes was 70%. 3. Levels of hepatocyte ethoxycoumarin-O-deethylase (ECOD) activity did not change following cryopreservation. The rate of decline of ECOD activity with time in culture was similar in freshly isolated and cryopreserved hepatocytes. 4. Hepatocytes isolated from three human livers were cryopreserved and recovered with viabilities similar to those obtained with the rat. A preliminary experiment also showed no loss of metabolic activity in human hepatocytes following cryopreservation.


A randomized clinical trial aimed at preventing poor psychosocial and glycemic outcomes in teens with type 1 diabetes (T1D).
Adolescents with type 1 diabetes have an increased risk for a variety of emotional and behavioral challenges as well as negative diabetes outcomes. This study was designed to compare the effectiveness of a depression-prevention, resilience promotion program with an advanced diabetes education program. Each program consisted of 9 group-based sessions. There were 264 adolescents enrolled in this multi-site randomized clinical trial. The primary outcomes were depressive symptoms and glycemic control; secondary outcomes included resilience skills, diabetes management and adherence, and diabetes-specific distress. The goal of the present paper is to describe the study design, the intervention, and the baseline characteristics of the sample. Preliminary data suggests that enrollment, randomization and retention were successful. Longitudinal follow-up and examination of mechanisms of action as they relate to psychosocial and glycemic outcomes will be explored in the future.


Enhanced tumor accumulation of sub-2 nm gold nanoclusters for cancer radiation therapy.
A new type of metabolizable and efficient radiosensitizers for cancer radiotherapy is presented by combining ultrasmall Au nanoclusters (NCs, <2 nm) with biocompatible coating ligands (glutathione, GSH). The new nanoconstruct (GSH-coated Au25 NCs) inherits attractive features of both the Au core (strong radiosensitizing effect) and GSH shell (good biocompatibility). It can preferentially accumulate in tumor via the improved EPR effect, which leads to strong enhancement for cancer radiotherapy. After the treatment, the small-sized GSH-Au25 NCs can be efficiently cleared by the kidney, minimizing any potential side effects due to the accumulation of Au25 NCs in the body.


Molecular cloning of a lobster G alpha(q) protein expressed in neurons of olfactory organ and brain.
We have isolated from an American lobster (Homarus americanus) olfactory organ cDNA library a clone, hG alpha(q), with >80% identity to mammalian and arthropod G alpha(q) sequences. In brain and olfactory organ, hG alpha(q) mRNA was expressed predominantly in neurons, including virtually all the neuronal cell body clusters of the brain. G alpha(q) protein was also expressed broadly, appearing on western blots as a single band of 46 kDa in brain, eyestalk, pereiopod, dactyl, tail muscle, olfactory organ, and aesthetasc hairs. These results suggest that hG alpha(q) plays a role in a wide variety of signal transduction events. Its presence in the olfactory aesthetasc hairs, which are almost pure preparations of the outer dendrites of the olfactory receptor neurons, the expression of a single hG alpha(q) mRNA species (6 kb) in the olfactory organ, and the localization of hG alpha(q) mRNA predominantly in the olfactory receptor neurons of the olfactory organ strongly suggest that one function of hG alpha(q) is to mediate olfactory transduction.


A decision-making tree for evaluating an esthetically compromised single dental implant.
To develop a comprehensive decision-making tree for evaluating mid-facial peri-implant soft tissue dehiscence in the esthetic zone and provide a systematic approach for assessing various clinical case scenarios, determining appropriate treatment strategies, and considering factors such as the need for soft tissue augmentation, prosthetic changes, or implant removal. This clinical decision tree illustrates numerous case scenarios with various esthetic complications around an esthetically compromised, but clinically healthy single implant and provides clinicians with possible solutions as a predictable map for horizontal and vertical soft tissue augmentation in order to manage different clinical circumstances. According to current evidence, the key to treating such esthetic complications is the use of an adequate pre-surgical prosthetic interdisciplinary approach with proper surgical techniques in order to optimize soft tissue dimensions and create better esthetic results. This may be accomplished through a purely surgical, combination of surgical and prosthetic, or purely prosthetic approaches. The present report describes a series of successfully treated peri-implant esthetic complication cases in accordance with the decision-making tree that the authors recommend in order to achieve better long-term esthetic outcomes. The combination of adequate pre-surgical prosthetic interdisciplinary collaboration and proper surgical technique is critical in the optimization of sufficient soft tissue dimensions and contributes to a more highly esthetic result. This study demonstrates a clinical decision-making tree to provide comprehensive, effective therapy of an esthetically compromised dental implant by using one of the following approaches: purely prosthetic, purely surgical, or a combination of surgical and prosthetic with or without abutment removal.


Outcome of bipolar electrocoagulation with lesionectomy in the treatment of epilepsy involving eloquent areas.
We have demonstrated previously that bipolar electrocoagulation on functional cortex (BCFC) is a safe and effective approach for epilepsy involving eloquent areas. Here, we report the results of BCFC with lesionectomy for patients with epileptogenic foci partially overlapping eloquent areas. Forty patients who had been treated with lesionectomy with BCFC were retrospectively reviewed with regard to seizure outcome and neurological deficits. Ten similar patients who had received lesionectomy with multiple subpial transections (MST) were examined as a control group. In the lesionectomy group with BCFC, Engel class I was achieved in 18 (45%) patients, class II in 8 (20%) patients, class III in 8 (20%) patients and class IV in 6 (15%) patients. Five (12.5%) patients developed mild hemiparesis and 1 (2.5%) patient mild sensory dysphasia. In the lesionectomy group with MST, Engel class I was achieved in 3 (30%) patients, class II in 2 (20%) patients, class III in 3 (30%) patients and class IV in 2 (20%) patients. Two (20%) patients developed mild hemiparesis and 1 (10%) patient moderate hemiparesis. All these complications recovered within 1-12 months. Compared with MST, the outcome of BCFC with lesionectomy is similar. But since MST leads to mechanical injury, while BCFC causes thermal injury, the complications of BCFC seem less severe.


Quantitative experimental assessment of the factors contributing to hearing loss in serous otitis media.
This study was designed to quantitatively assess the contribution of various factors to the conductive hearing loss in otitis media. In the conductive hearing loss seen in cases of serous otitis media, various volumes of fluid of different viscosities along with subatmospheric (negative) pressure are found in the middle ear. To evaluate the contribution of each of these factors to hearing loss, various volumes of saline, whole blood, or glycerol were applied to the open middle ear cavity of guinea pigs for short periods of time and auditory function was evaluated by recording the threshold of auditory nerve-brainstem evoked responses. In some of the saline experiments, the bulla cavity was also sealed, allowing a subatmospheric (negative) pressure to develop in the cavity as water was osmotically absorbed because of the gradient in colloid osmotic pressure between saline and blood in the vessels lining the middle ear cavity. In other experiments, a thoracic drainage system was connected to the middle ear cavity to induce desired negative middle ear pressures. The degree of hearing loss increased as larger volumes of fluid were introduced into the middle ear, reaching a maximum of 15 to 16 dB. There was no difference in the degree of hearing loss induced by saline or by fluids with viscosities up to 1,000 times greater than that of water (glycerol). A subatmospheric pressure in the middle ear contributed only a small additional (1-2 dB) threshold elevation. The major factor contributing to hearing loss in serous otitis media is the volume of fluid in the middle ear, irrespective of its viscosity. The contribution of negative middle ear pressure is much smaller.


Insight into the molecular mechanism of miR-192 regulating Escherichia coli resistance in piglets.
MicroRNAs (miRNAs) have important roles in many cellular processes, including cell proliferation, growth and development, and disease control. Previous study demonstrated that the expression of two highly homologous miRNAs (miR-192 and miR-215) was up-regulated in weaned piglets with Escherichia coli F18 infection. However, the potential molecular mechanism of miR-192 in regulating E. coli infection remains unclear in pigs. In the present study, we analyzed the relationship between level of miR-192 and degree of E. coli resistance using transcription activator-like effector nuclease (TALEN), in vitro bacterial adhesion assays, and target genes research. A TALEN expression vector that specifically recognizes the pig miR-192 was constructed and then monoclonal epithelial cells defective in miR-192 were established. We found that miR-192 knockout led to enhance the adhesion ability of the E. coli strains F18ab, F18ac and K88ac, meanwhile increase the expression of target genes (DLG5 and ALCAM) by qPCR and Western blotting analysis. The results suggested that miR-192 and its key target genes (DLG5 and ALCAM) could have a key role in E. coli infection. Based on our findings, we propose that further investigation of miR-192 function is likely to lead to insights into the molecular mechanisms of E. coli infection.


Pro-angiogenic cytokines and their role in tumor angiogenesis.
The development of solid tumors depends upon an adequate supply of blood. This can be achieved by way of co-option of preexisting blood vessels and by the induction of angiogenesis. During the past 30 years, tumor angiogenesis had been found to play a crucial role in the progression of solid tumors. Tumor angiogenesis was found to be induced by a variety of pro-angiogenic cytokines of which the best characterized is vascular endothelial growth factor (VEGF). Indeed, the first FDA approved anti-angiogenic drug for the treatment of cancer is Avastin, a neutralizing antibody directed against VEGF. This review focuses on cytokines which have been reported to induce tumor angiogenesis.


Methylation of the ER-alpha Promoter Is Influenced by its Ligand Estrogen in Osteosarcoma Cells SAOS-2 In Vitro.
The aggressive fast-growing osteosarcoma is the most common primary malignant bone tumor. The relevance of estrogen as a key player in bone metabolism and bone tumor is well-known. At the molecular level, estrogen activates the estrogen receptor α (ERα) as a natural ligand of this receptor. ERα acts as a transcription factor by binding to the "estrogen response element" (ERE) and regulates the expression of a various number of genes. Epigenetic processes, e.g. the methylation of the "cytosine-phosphatidyl-guanine (CpG) islands" can change the transcription of target genes and subsequently the protein expression. As DNA methylation is generally associated with gene transcription repression, up until now little is known about the ERα methylation in osteosarcoma cells. The aim of the present pilot study was to evaluate the methylation status of ERα in osteosarcoma cells SAOS-2 and MG 63 after stimulation with estrogen. SAOS-2 and MG 63 cells were cultured in DMEM. After treatment with 10 nmol estrogen (E2) for 24 h, the expression of ERα was detected by immunocytochemistry (ICC). As controls we used untreated cells. Staining was evaluated semi-quantitatively by the immunoreactive score of Remmele and Stegner (IRS). To determine mRNA gene expression, extracted RNA was transcribed into c-DNA and a quantitative real-time-PCR (qRT-PCR) was carried out. The semi quantitative evaluation of the ERα mRNA was based on the 2(-ΔΔct) method using untreated cells as reference control. One microgram of each extracted genomic DNA sample was converted with bisulfite and a real-time methylation-specific PCR (rt-MSP) was performed. The estrogen-stimulated SAOS-2 cells showed a significant increase of ERα expression. A 7-fold up-regulation of ERα mRNA confirmed the results of immunocytochemistry. Methylation of the ERα promoter was not detected in treated cells. In contrast, we identified methylation of the ERα promoters in untreated cells. The staining of MG 63 cells showed a weak gain of ERα expression in the stimulated cells, as well as a weak increase of the ER-α mRNA (2-fold). Methylation of the ERα promoters was not detectable in either treated or untreated cells. The methylation status of ERα in osteosarcoma cells is affected by estrogen. These findings indicate that epigenetic changes of genomic DNA regulate ERα synthesis. Taken together, our results suggest that SAOS-2 cells can be an interesting model for further investigating ERα synthesis. In addition, the evaluation of ERα methylation in osteosarcoma specimens is in progress.


ScrumPy: metabolic modelling with Python.
ScrumPy is a software package used for the definition and analysis of metabolic models. It is written using the Python programming language that is also used as a user interface. ScrumPy has features for both kinetic and structural modelling, but the emphasis is on structural modelling and those features of most relevance to analysis of large (genome-scale) models. The aim is at describing ScrumPy's functionality to readers with some knowledge of metabolic modelling, but implementation, programming and other computational details are omitted. ScrumPy is released under the Gnu Public Licence, and available for download from http://mudshark.brookes.ac.uk/ ScrumPy.


Locally infiltrative inflammatory fibroid polyp of the ileum: report of a case showing transmural proliferation.
Morphologically, an inflammatory fibroid polyp (IFP) is usually centred in the submucosa. Extension of an IFP to the subserosa with destruction of the muscularis propria is exceedingly rare. Herein, we describe a 70-year-old woman who presented with right lower abdominal pain but was finally diagnosed with an IFP. Contrast-enhanced computed tomography revealed a target-like structure with a hypovascular mass at the leading edge, which was consistent with intussusception due to a tumour. Following surgery, the resected specimen displayed a mass measuring 4 × 3 × 3 cm that was protruding into the lumen. Microscopically, the mass was centred in the submucosa, extending up to the mucosal surface and down to the subserosa and serosa. The muscularis mucosae and muscularis propria were destroyed focally. A PDGFRA gene mutation in exon 2 (1837_1851 del) that was found in this case, as well as a highly infiltrative growth pattern, strongly supported the neoplastic nature of IFP.


Twice Silenced: The Underreporting of Child Sexual Abuse in Orthodox Jewish Communities.
Child sexual abuse remains an underreported crime throughout the world, despite extensive research and resources dedicated both to improving investigative techniques and helping children disclose their experiences. The discovery of rampant cover-ups within the Catholic Church has exposed some of the ways religious and cultural issues can impede reporting to authorities. This article examines specific factors that contribute to the underreporting of child sexual abuse within Orthodox Jewish communities. It also explores ways in which these communities have handled child sexual abuse reporting in the past and describes recent progress. Implications are offered for CSA prevention, detection, and recovery in Orthodox Jewish communities as well as other minority religious groups.


Heritability of hair whorl position on the forehead in Konik horses.
There are studies on the relationship between the position and shape of hair whorls on bovine forehead and phenotypic traits. According to anecdotal beliefs by horse users and handlers, temperamental traits may be related to the position of hair whorls in horses. No previous research on the mechanisms of inheritance of hair whorls has been performed, so the aim of the present study was to determine the heritability of the position of the hair whorl on the forehead of Konik horses. The horses (n = 362) were classified into five groups based on the whorl position on forehead with respect to the top and bottom eye lines. The estimated heritability of hair whorl position was 0.753 (SE = 0.056). Heritability adjusted for the discontinuity of the trait was 0.836. The results show that hair whorl position in Konik Polski horses is highly heritable. The possible relationship between position of hair whorls on the forehead and other morphological traits needs further research and should be interpreted with caution.


Buckling-Induced Kirigami.
We investigate the mechanical response of thin sheets perforated with a square array of mutually orthogonal cuts, which leaves a network of squares connected by small ligaments. Our combined analytical, experimental and numerical results indicate that under uniaxial tension the ligaments buckle out of plane, inducing the formation of 3D patterns whose morphology is controlled by the load direction. We also find that by largely stretching the buckled perforated sheets, plastic strains develop in the ligaments. This gives rise to the formation of kirigami sheets comprising periodic distribution of cuts and permanent folds. As such, the proposed buckling-induced pop-up strategy points to a simple route for manufacturing complex morphable structures out of flat perforated sheets.


Comparison of operating field sterility in open versus minimally invasive microdiscectomies of the lumbar spine.
Postoperative wound infection is a preventable risk that can lead to significant adverse outcomes and increased cost of care. Minimally invasive surgeries (MIS) have been found to have lower rates of postoperative infection compared with the traditional approach. To assess if the reported difference is related to intraoperative contamination or to other factors, we assessed the surgical field for sterility. We compared 10 MIS versus 10 traditional microdiscectomies. Swabs of the operating field were obtained before and after the procedure from multiple sites in the operating room. Positive and negative controls were taken of the skin immediately before and after preparation of the incision site. All swabs were plated out on Columbia blood agar plates and grown for 48 hours. Colony counting was performed to determine growth. There was no statistically significant difference in the colony counts of swab sites in traditional microdiscectomies compared with MIS microdiscectomies. There was no significant contamination of the operating field using either approach. In this prospective study, we found that there was no significant difference in bacterial counts in swabs of operative sites in either traditional or MIS microdiscectomies, suggesting that the decreased rate of postoperative infection in the reported literature for MIS cases may be related to other factors, such as patient selection and/or postoperative care.


Hydration free energy difference of acetone, acetamide, and urea.
The hydration free energy and hydration entropy difference of urea and acetone, and of acetamide and acetone have been calculated both by free energy perturbation and by the method of thermodynamic integration. The obtained results show a striking asymmetry between the thermodynamic changes accompanying the replacement of the first and second CH(3) group of acetone by NH(2). Thus, the first CH(3)NH(2) exchange is found to lead to an about 10 kJmol decrease in the energy, 8 kJmol decrease in the Helmholtz free energy, and 5-10 Jmol K decrease in the entropy of hydration, while similar values accompanying the second CH(3)NH(2) exchange are found to be about -65 kJmol, -35 kJmol and -100 Jmol K, respectively. These results indicate that the two NH(2) groups of the urea molecule have a strong synergetic effect on the thermodynamics of the hydration of urea. The fact that the replacement of the two CH(3) groups of acetone by NH(2) leads to a strong decrease in the hydration entropy indicates that urea clearly has an ordering effect on nearby water.


Biological roles of interferon tau (IFNT) and type I IFN receptors in elongation of the ovine conceptus.
Interferon tau (IFNT) is produced by the elongating conceptus in ruminants and is the maternal recognition of the pregnancy signal. Available evidence supports the idea that IFNT acts in a paracrine and autocrine manner to modulate expression of genes in the endometrium and trophectoderm, respectively, which promote conceptus elongation. The actions of IFNT are mediated by the interferon (alpha and beta) receptor (IFNAR), which consists of two subunits, IFNAR1 and IFNAR2. To test the hypothesis that IFNT and its receptor have biological roles in conceptus elongation, an in vivo loss-of-function study was conducted by inhibiting IFNT or IFNAR1/2 mRNA translation in the trophectoderm of the ovine conceptus using morpholino antisense oligonucleotides (MAO) delivered via osmotic pumps from Days 8 to 14 postmating. Elongating, filamentous type conceptuses were recovered from Day 14 ewes receiving a control morpholino or IFNAR MAOs. In contrast, severely growth-retarded and malformed conceptuses were recovered from IFNT MAO-infused ewes. Those conceptuses contained abnormal trophectoderm cells that were apoptotic based on terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling analyses. IFNT concentrations were reduced in the uterine lumen of IFNT MAO-infused ewes as was the expression of classical Type I IFN-stimulated genes in the endometrium. IFNT concentrations were also lower in the uterine lumen of IFNAR1/2 MAO-infused ewes. These studies provide in vivo evidence that IFNT is a critical regulator of conceptus elongation and that its embryotrophic actions are primarily mediated by paracrine effects on the endometrium.


Neovascular Glaucoma as a Presenting Sign of Catastrophic Antiphospholipid Syndrome with a "Catastrophic" Heart Valve Finding.
We aimed to describe a case of neovascular glaucoma (NVG) as a first presenting sign of catastrophic antiphospholipid syndrome (CAPS) with heart valve aseptic vegetations known as Libman-Sacks endocarditis. A 39-year-old man was referred for left eye decreased visual acuity and pain, upon examination left eye high intraocular pressure; rubeosis iridis of both eyes (BE); and prominent retinal ischemia. Clinical and fluorescein angiography findings established the diagnosis of left eye NVG with vaso-occlusive disease in BE. Magnetic resonance imaging of the head showed widespread ischemic lesions and hemorrhagic foci. The transesophageal echocardiogram showed 2 big mitral valve lesions consistent with the diagnosis of Libman-Sacks endocarditis. Laboratory and clinical diagnosis of CAPS and suspected SLE was confirmed, and treatment with anticoagulants and IV steroids was initiated. This case demonstrates that severe vaso-occlusive retinopathy with severe brain ischemia should raise the suspicion of systemic autoimmune pro-coagulative diseases with heart valve aseptic vegetations.


TRB3 mediates advanced glycation end product-induced apoptosis of pancreatic β-cells through the protein kinase C β pathway.
Advanced glycation end products (AGEs), which accumulate in the body during the development of diabetes, may be one of the factors leading to pancreatic β-cell failure and reduced β-cell mass. However, the mechanisms responsible for AGE‑induced apoptosis remain unclear. This study identified the role and mechanisms of action of tribbles homolog 3 (TRB3) in AGE-induced β-cell oxidative damage and apoptosis. Rat insulinoma cells (INS-1) were treated with 200 µg/ml AGEs for 48 h, and cell apoptosis was then detected by TUNEL staining and flow cytometry. The level of intracellular reactive oxygen species (ROS) was measured by a fluorescence assay. The expression levels of receptor of AGEs (RAGE), TRB3, protein kinase C β2 (PKCβ2) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) were evaluated by RT-qPCR and western blot analysis. siRNA was used to knockdown TRB3 expression through lipofection, followed by an analysis of the effects of TRB3 on PKCβ2 and NOX4. Furthermore, the PKCβ2-specific inhibitor, LY333531, was used to analyze the effects of PKCβ2 on ROS levels and apoptosis. We found that AGEs induced the apoptosis of INS-1 cells and upregulated RAGE and TRB3 expression. AGEs also increased ROS levels in β-cells. Following the knockdown of TRB3, the AGE-induced apoptosis and intracellular ROS levels were significantly decreased, suggesting that TRB3 mediated AGE-induced apoptosis. Further experiments demonstrated that the knockdown of TRB3 decreased the PKCβ2 and NOX4 expression levels. When TRB3 was knocked down, the cells expressed decreased levels of PKCβ2 and NOX4. The PKCβ2‑specific inhibitor, LY333531, also reduced AGE-induced apoptosis and intracellular ROS levels. Taken together, our data suggest that TRB3 mediates AGE-induced oxidative injury in β-cells through the PKCβ2 pathway.


Tissue-specific expression of mast cell granule serine proteinases and their role in inflammation in the lung and gut.
Serine proteinases with trypsin-like (tryptase) and chymotrypsin-like (chymase) properties are major constituents of mast cell granules. Several tetrameric tryptases with differing specificities have been characterized in humans, but only a single chymase. In other species there are larger families of chymases with distinct and narrow proteolytic specificities. Expression of chymases and tryptases varies between tissues. Human pulmonary and gastrointestinal mast cells express chymase at lower levels than tryptase, whereas rodent and ruminant gastrointestinal mast cells express uniquely mucosa-specific chymases. Local and systemic release of chymases and tryptases can be quantified by immunoassay, providing highly specific markers of mast cell activation. The expression and constitutive extracellular secretion of the mucosa-specific chymase, mouse mast cell proteinase-1 (mMCP-1), is regulated by transforming growth factor-beta1 (TGF-beta1) in vitro, but it is not clear how the differential expression of chymases and tryptases is regulated in other species. Few native inhibitors have been identified for tryptases but the tetramers dissociate into inactive subunits in the absence of heparin. Chymases are variably inhibited by plasma proteinase inhibitors and by secretory leucocyte protease inhibitor (SLPI) that is expressed in the airways. Tryptases and chymases promote vascular permeability via indirect and possibly direct mechanisms. They contribute to tissue remodelling through selective proteolysis of matrix proteins and through activation of proteinase-activated receptors and of matrix metalloproteinases. Chymase may modulate vascular tissues through its ability to process angiotensin-I to angiotensin-II. Mucosa-specific chymases promote epithelial permeability and are involved in the immune expulsion of intestinal nematodes. Importantly, granule proteinases released extracellularly contribute to the recruitment of inflammatory cells and may thus be involved in innate responses to infection.


Multiplex family with GAD65-Abs neurologic syndromes.
Neurologic autoimmune syndromes associated with anti-glutamate acid decarboxylase 65 antibodies (GAD65-Abs) are rare and mostly sporadic. We describe a niece and her aunt with GAD65-Abs neurologic syndromes. High-resolution HLA typing of Class I and Class II alleles was performed using next-generation sequencing. The proband had cerebellar ataxia and probable limbic encephalitis features, whereas her niece had stiff-person syndrome. Both had a high titer of GAD65-Abs in serum and CSF and showed signs of inflammation in CSF. Both affected members carried the same rare recombinant DRB1*15:01:01∼DQA1*01:02:01∼DQB1*05:02:01 haplotype, which may or may not be involved in disease susceptibility. Of interest, other unaffected members of the family either had the same HLA haplotype but normal serum GAD65-Abs or had different HLA types but a high titer of serum GAD65-Abs without neurologic symptoms, suggesting cumulative effects. This unique association strengthens the concept that hereditary factors, possibly including specific HLA haplotypes, play a role in neurologic syndromes associated with GAD65-Abs.


Outcome of recurrent pterygium with intraoperative 0.02% mitomycin C and free flap limbal conjunctival autograft.
To determine the re-recurrence and the postoperative complications in recurrent pterygium cases treated by intraoperative 0.02% Mitomycin C (MMC) and conjunctival autograft (CAG). Quasi-experimental. Department of Ophthalmology, Liaquat University of Medical and Health Sciences, Jamshoro, Hyderabad and Chandka Medical College Hospital, Larkana, from January to December 2010. Cases with recurrent pterygium were included in this study. Cases with history of first time pterygium and pterygium with conjunctival scarring, dry eye, glaucoma and vitreoretinal disease were excluded. After topical and subconjunctival anaesthesia, pterygium was excised in single piece. Intraoperative 0.02% MMC was applied on bare sclera for 2 minutes. CAG was excised from supero-temporal conjunctiva, and implanted on bare sclera. All cases were followed-up for 6 months. Re-recurrence was defined as postoperative fibrovascular re-growth of 1.0 mm or more crossing the corneo-scleral limbus. Data was analysed as descriptive statistics. A total of 65 eyes of 65 cases were studied having mean age of 43.26 ± 12.81 years. Among those, 41 (63.1%) were males and 48 (73.8%) belonged to rural area. The size of pterygium on cornea was 2-3 mm in 44 (67.7%) cases and 4-5 mm in 21 (32.3%) cases. Re-recurrence of pterygium was seen in 3 (4.6%) cases. Postoperative complications included conjunctival granuloma in 2 (3.1%) cases, graft necrosis in 2 (3.1%) cases, graft oedema in 3 (4.6%) cases and graft displacement in 3 (4.6%) cases. The intraoperative application of 0.02% MMC with CAG markedly reduces the risk of re-recurrence of pterygium and postoperative complications.


Study on vapor-liquid equilibria and surface tensions for nonpolar fluids by renormalization group theory and density gradient theory.
An equation of state (EOS) applicable for both the uniform and nonuniform fluids is established by using the density-gradient theory (DGT). In the bulk phases, the EOS reduces to statistical associating fluid theory (SAFT). By combining the EOS with the renormalization group theory (RGT), the vapor-liquid-phase equilibria and surface tensions for 10 nonpolar chainlike fluids are investigated from low temperature up to the critical point. The obtained results agree well with the experimental data.


Sea urchin injuries: a review and clinical approach algorithm.
Sea urchin injuries (SUIs) are among the most common marine injuries; however, there are no guidelines to dictate appropriate workup and treatment. Complications vary significantly depending on the extent of injury, site of injury and elapsed time to medical attention. Timely and appropriate management can prevent long-term morbidity despite the deceptively innocuous appearance of these injuries. We review the English literature on SUIs and offer an algorithm to aid in the management of affected patients. We found that superficially retained spines may trigger a local granulomatous inflammatory response, while spines retained near deep structures may induce sea urchin arthritis, inflammatory tenosynovitis, among other delayed complications. Therefore, we recommend immediate inactivation of pro-inflammatory compounds by hot water soaks at the time of injury, followed by extraction of all spines by a physician. Imaging is a valuable component of risk stratification to determine depth and location of spines, which guides selection of appropriate extraction technique to prevent long-term morbidity.


Variation Quality and Kinetic Parameter of Commercial n-3 PUFA-Rich Oil during Oxidation via Rancimat.
Different biological sources of n-3 polyunsaturated fatty acids (n-3 PUFA) in mainstream commercial products include algae and fish. Lipid oxidation in n-3 PUFA-rich oil is the most important cause of its deterioration. We investigated the kinetic parameters of n-3 PUFA-rich oil during oxidation via Rancimat (at a temperature range of 70~100 °C). This was done on the basis of the Arrhenius equation, which indicates that the activation energies (Ea) for oxidative stability are 82.84-96.98 KJ/mol. The chemical substrates of different oxidative levels resulting from oxidation via Rancimat at 80 °C were evaluated. At the initiation of oxidation, the tocopherols in the oil degraded very quickly, resulting in diminished protection against further oxidation. Then, the degradation of the fatty acids with n-3 PUFA-rich oil was evident because of decreased levels of PUFA along with increased levels of saturated fatty acids (SFA). The quality deterioration from n-3 PUFA-rich oil at the various oxidative levels was analyzed chemometrically. The anisidine value (p-AV, r: 0.92) and total oxidation value (TOTOX, r: 0.91) exhibited a good linear relationship in a principal component analysis (PCA), while oxidative change and a significant quality change to the induction period (IP) were detected through an agglomerative hierarchical cluster (AHC) analysis.


Adjuvant radiation in the TORS era: Is there a benefit to omitting the tumor bed?
Trans-oral robotic surgery (TORS) followed by adjuvant radiation therapy (RT) is becoming popular in treating node-positive tonsillar squamous cell carcinoma (SCC). One potential benefit is eliminating irradiation of the primary site when the tumor is widely resected, and targeting the neck only. This study assessed whether omitting the tumor bed provides any dosimetric or clinical advantage. We identified 21 patients with primary tonsillar SCC that were treated with TORS followed by RT to the ipsilateral neck and primary (N+P) site. We subsequently replanned each case to allow target coverage to the neck only, excluding the tumor bed. For each case, we created 2 plans: neck only (NO) and N+P. Additionally, we identified patients (n = 7) with primary SCC tonsil who did receive NO irradiation to compare their dosimetry and toxicity profile with those treated to both the primary site and ipsilateral neck. Contralateral neck radiation was not administered in any of these cases. Despite excluding the primary site in the NO plans, the mean dose to the tumor bed was high: 53.9 Gy. The only significant differences in omitting the primary site were lower doses to the oral cavity, 29.8 Gy in NO plan versus 34.6 Gy in N+P plan, P = .002, and superior constrictors, 42.9 Gy in NO plan versus 46.1 Gy in N+P plan, P = .01. No appreciable differences in toxicities were noted in the limited cohort treated with NO irradiation. Sparing the primary site after TORS in node-positive tonsillar SCC does not appear to provide any significant dosimetric or clinical advantage. Furthermore, the tumor bed receives a significant but potentially subtherapeutic dose, limiting options for irradiation in a salvage setting. At this time, we do not recommend omitting the tumor bed from the radiation target volume following TORS.


Fatty acid synthase as a potential therapeutic target in cancer.
Fatty acid synthase (FASN) is a key enzyme involved in neoplastic lipogenesis. Overexpression of FASN is common in many cancers, and accumulating evidence suggests that it is a metabolic oncogene with an important role in tumor growth and survival, making it an attractive target for cancer therapy. Early small-molecule FASN inhibitors such as cerulenin, C75 and orlistat have been shown to induce apoptosis in several cancer cell lines and to induce tumor growth delay in several cancer xenograft models but their mechanism is still not well understood. These molecules suffer from pharmacological limitations and weight loss as a side effect that prevent their development as systemic drugs. Several potent inhibitors have recently been reported that may help to unravel and exploit the full potential of FASN as a target for cancer therapy in the near future. Furthermore, novel sources of FASN inhibitors, such as green tea and dietary soy, make both dietary manipulation and chemoprevention potential alternative modes of therapy in the future.


Congenital intractable diarrhea with possible defective crypt regeneration: report of a case.
A male infant, born uneventfully from a consanguinous marriage, presented with intractable watery diarrhea from his third day of life, with subsequent malnutrition and failure to thrive. He received central parenteral nutrition beginning at three months of age after a poor response to a semielemental diet and peripheral parenteral nutrition. He was totally dependent on central parenteral nutrition thereafter. Although diarrhea disappeared with strict bowel rest, intolerance to minimal enteral feedings persisted throughout his 2 years 4 months of life. Investigations including stool examinations and repeated cultures, immune function studies, radiologic studies of the small bowel and screening for galactosemia and cystic fibrosis could not demonstrate a specific cause for the diarrhea. Repeated small intestinal biopsies at 1 month, 4 months and 1 year 5 months of age showed persistent villous atrophy with crypt hypoplasia and a low crypt mitotic index. Electron microscopic examination revealed normal-appearing microvilli. This child may have had a congenital enteropathy due to an inborn crypt regeneration defect causing lifelong intolerance to enteral feedings.


Radiotherapy preceded by multidrug chemotherapy in carcinoma of the esophagus: a pilot study of the Radiation Therapy Oncology Group.
This is a report on R.T.O.G. #77-07, a phase II pilot study aimed at determining the toxicity, primary tumor control, and survival achieved with a combination of triple-drug chemotherapy prior to radiotherapy in carcinoma of the esophagus. The drugs used were vincristine, bleomycin, and methotrexate. A total of 26 cases were registered, one of which died during chemotherapy; 11 had only one chemotherapy course, and 14 had two chemotherapy courses as planned. Drug toxicity could be evaluated in 23 patients: one died from liver damage secondary to chemotherapy effect (4%), two had nausea and vomiting (9%), one had weakness and skin rash (4%), and one had fever and vomiting (4%). There was no complete tumor response to chemotherapy in 22 evaluable cases; 12 of 22 (55%) showed some measurable tumor reduction. Radiation toxicity could be evaluated in 25 patients: 1 of 25 (4%) developed clinical pericarditis, and 2 of 25 (8%) developed severe esophagitis. A complete response to radiotherapy was observed in 15 of 25 (60%) patients; 7 of 25 (28%) showed a partial response; and 3 of 25 (12%) had no measurable tumor reduction. The median survival in 25 evaluable cases was 22 months. In 11 patients who received a single course of chemotherapy, the median survival was 19 months, while in 14 patients who received two courses of chemotherapy, the median survival was only 9 months. However, this difference is not statistically significant.


Computer-based education for patients with cancer at Latino border hospital.
There is a paucity of relevant educational material for Latino patients with cancer. Education interventions used for non-Latinos must be assessed for relevancy for Latino patients. This study examined the use and usefulness of computer-based education interventions among Latinos. It was conducted using the CancerHelp software survey in a Texas institution on the US-Mexico border. Nonparametric test statistics were used to test the hypothesized relationships and the usefulness of CancerHelp software. The following statistic hypotheses were not rejected: (1) CancerHelp use is independent of age group, (2) use of CancerHelp is equally distributed among patients and family members, (3) among Latinos, CancerHelp use is independent of education, and (4) CancerHelp usefulness for patients with cancer is independent of ethnicity.


Regression of nonalcoholic fatty liver disease is associated with reduced risk of incident diabetes: A longitudinal cohort study.
Non-alcoholic fatty liver disease (NAFLD) is potentially reversible. However, whether improvement of NAFLD leads to clinical benefits remains uncertain. We investigated the association between regression of NAFLD and the risk of incident diabetes in a longitudinal way. A cohort of 11,260 adults who had NAFLD at in an initial exam, had the second evaluation for NAFLD status at 1~2 years from an initial exam were followed up for incident diabetes from 2001 and 2016. NAFLD was diagnosed with abdominal ultrasound. At baseline, NAFLD was regressed in 2,559 participants (22.7%). During 51,388 person-years of follow-up (median 4 years), 1,768 participants developed diabetes. The fully adjusted hazard ratio (HR) for incident diabetes in participants with regressed NAFLD compared to those with persistent NAFLD was 0.81 [95% confidence interval (CI) 0.72-0.92]. When assessed by NAFLD severity, among participants with a low NAFLD fibrosis score (NFS) (< -1.455), participants with regressed NAFLD had a lower risk of incident diabetes than those with persistent NAFLD (HR 0.77, 95% CI 0.68-0.88). However, in participants with an intermediate to high NFS (≥ -1.455), the risk of incident diabetes was not different between NAFLD regression and persistence groups (HR 1.12, 95% CI 0.82-1.51). Regression of NAFLD was associated with decreased risk of incident diabetes compared to persistent NAFLD. However, the benefit was evident only for NAFLD patients with low NFS. This suggests that early intervention for NAFLD, before advanced fibrosis is present, may maximize the metabolic benefit from NAFLD regression.


Quantifying exposure to diagnostic medical radiation in patients with inflammatory bowel disease: are we contributing to malignancy?
While potential risks of diagnostic medical radiation are acknowledged, actual exposure of patients in routine clinical practice is poorly documented. To quantify such exposure to vulnerable abdominal organs in patients with inflammatory bowel disease who are already at risk of intestinal cancer. All incidences of exposure to diagnostic medical radiation were documented in a consecutive series of 100 patients with inflammatory bowel disease (62 Crohn's disease, 37 ulcerative colitis, 1 indeterminate colitis) attending a hospital-based clinic. Total effective dose (mSv) was calculated using published tables. Predictors of high or no irradiation were evaluated by multivariate logistic regression analysis. Thirteen patients had no documented diagnostic irradiation. Twenty-three patients received an effective dose greater than 25 mSv. An at-risk effective dose >50 mSv was received by 11 patients. Dosage was higher in patients with Crohn's disease than ulcerative colitis (P = 0.02) and in patients undergoing surgery (P = 0.004). However, no predictive factors for high radiation dosage or for no exposure were identified. At-risk irradiation from diagnostic medical radiation is common in patients with inflammatory bowel disease, and might potentially contribute to the elevated risk of intra-abdominal and other cancers. The level of irradiation should be considered in clinical decisions regarding abdominal imaging.


Health-related quality of life in long term breast cancer survivors treated with breast conserving therapy: impact of age at therapy.
Aim was to compare the functional status in long-term breast cancer survivors related to age at diagnosis and to asses the effects of adjuvant therapy on health-related quality of life (HRQoL). Data were obtained from 370 patients after breast conserving therapy (BCT) at follow-up (F/U) visit. The self-administered EORTC QoL questionnaire (C30) and the breast module (QLQ-BR23) measuring global health, global QoL, physical, role, emotional, cognitive and social functioning, body image, sexual function, future perspective and arm/breast symptoms were used. Patients were grouped according to F/U (12 years, 7 years), age at therapy (< 50 years, 50-65 years, >65 years) and adjuvant treatment (none, chemotherapy, hormone replacement therapy). The global HRQoL was increased in patients with longer F/U (p < 0.01). Physical functioning, role functioning and sexual functioning were decreased in patients being older at therapy (> 65 years, p < 0.01). Increased arm symptoms were noticed in older women at longer follow-up. Younger women at therapy complained financial difficulties at follow-up (p < 0.006). Adjuvant chemotherapy and hormone replacement therapy did not affect physical and mental functioning. Women of different age treated with BCT for breast cancer should be considered at a different risk for HRQoL disturbance at long term F/U in both, physical and psychological dimensions. In clinical practice, specific identification of those women with negative impact of diagnosis and treatment on long term HRQoL would help for targeted interventions. In clinical studies focusing on HRQoL, the compared groups need to be age-adjusted.


Elucidation of the role of Grr1p in glucose sensing by Saccharomyces cerevisiae through genome-wide transcription analysis.
The role of Grr1p in glucose sensing in Saccharomyces cerevisiae was elucidated through genome-wide transcription analysis. From triplicate analysis of a strain with deletion of the GRR1-gene from the genome and an isogenic reference strain, 68 genes were identified to have significantly altered expression using a Student's t-test with Bonferroni correction. These 68 genes were widely distributed across different parts of the cellular metabolism and GRR1-deletion is therefore concluded to result in polytrophic effects, indicating multiple roles for Grr1p. Using a less conservative statistical test, namely the SAM test, 232 genes were identified as having significantly altered expression, and also these genes were widely distributed across different parts of the cellular metabolism. Promoter analyses on a genome-wide scale and on the genes with significant changes revealed an over-representation of DNA-binding motifs for the transcriptional regulators Mig1p and Rgt1p in the promoter region of the significantly altered genes, indicating that Grr1p plays an important role in the regulatory pathways that ultimately lead to transcriptional regulation by each of the components Mig1p and Rgt1p.


Experimental and Mathematical Optimization of a Pooling Test for Detection of SARS-CoV-2 in a Population with Low Viral Load.
A pooling test is a useful tool for mass screening of coronavirus disease 2019 (COVID-19) in the pandemic era. We aimed to optimize a simple two-step pooling test by estimating the optimal pool size using experimental and mathematical validation. Experimental pools were created by mixing one positive respiratory sample with various numbers of negative samples. We selected positive samples with cycle threshold (Ct) values greater than 32 to validate the efficiency of the pooling test assuming a high likelihood of false-negative results due to low viral loads. The positivities of the experimental pools were investigated with a single reverse-transcription polymerase chain reaction (RT-PCR) using the U-TOP™ COVID-19 Detection Kit Plus (Seasun Biomaterials, Daejeon, Korea). We used the Dorfman equation to calculate the optimal size of a pooling test mathematically. Viral RNA could be detected in a pool with a size up to 11, even if the Ct value of a positive sample was about 35. The Dorfman equation showed that the optimal number of samples in a pool was 11 when the prevalence was assumed to be 0.66% based on the test positivity in Daejeon, Korea from April 1, 2020 to November 10, 2020. The efficiency of the pooling test was 6.2, which can save 83.9 of 100 individual tests. Eleven samples in a pool were validated optimal experimentally assuming a prevalence of 0.66%. The pool size needs modification as the pandemic progresses; thus, the prevalence should be carefully estimated before pooling tests are conducted.


The history of treatment of congenital clubfoot at the Royal Liverpool Children's Hospital: improvement of results by early extensive posteromedial release.
The results of treatment in 77 clubfeet were assessed after an average follow-up of 8 years. In those patients with surgical treatment it was found that the Turco posteromedial release within the first 3 months of life resulted in 13 of 16 satisfactory feet. In comparison, only five of 21 feet had satisfactory results when the average age of operation was delayed to 3.7 years and a less extensive release performed. At follow-up, abnormal radiological findings were not found in the early operated cases. The average talocalcaneal index was 47 degrees. The late operated feet showed major abnormal X-ray film findings, and their average index was 38 degrees. Good functional results, however, often coexisted with abnormal radiological findings in the midfoot. Abnormal findings of the hindfoot were always associated with unsatisfactory results. We conclude that the use of early (3 months) Turco posteromedial release can offer excellent results in treating severe resistant clubfeet.


Genetic tests of ancient asexuality in root knot nematodes reveal recent hybrid origins.
The existence of "ancient asexuals", taxa that have persisted for long periods of evolutionary history without sexual recombination, is both controversial and important for our understanding of the evolution and maintenance of sexual reproduction. A lack of sex has consequences not only for the ecology of the asexual organism but also for its genome. Several genetic signatures are predicted from long-term asexual (apomictic) reproduction including (i) large "allelic" sequence divergence (ii) lack of phylogenetic clustering of "alleles" within morphological species and (iii) decay and loss of genes specific to meiosis and sexual reproduction. These genetic signatures can be hard to assess since it is difficult to demonstrate the allelic nature of very divergent sequences, divergence levels may be complicated by processes such as inter-specific hybridization, and genes may have secondary roles unrelated to sexual reproduction. Apomictic species of Meloidogyne root knot nematodes have been suggested previously to be ancient asexuals. Their relatives reproduce by meiotic parthenogenesis or facultative sexuality, which in combination with the abundance of nematode genomic sequence data, makes them a powerful system in which to study the consequences of reproductive mode on genomic divergence. Here, sequences from nuclear protein-coding genes are used to demonstrate that the first two predictions of ancient asexuality are found within the apomictic root knot nematodes. Alleles are more divergent in the apomictic taxa than in those species exhibiting recombination and do not group phylogenetically according to recognized species. In contrast some nuclear alleles, and mtDNA, are almost identical across species. Sequencing of Major Sperm Protein, a gamete-specific gene, from both meiotic and ameiotic species reveals no increase in evolutionary rate nor change in substitution pattern in the apomictic taxa, indicating that the locus has been maintained by selection. The data strongly suggests the tropical root knot nematode apomicts have a recent origin and are not anciently asexual. The results support that interspecific hybridization has been involved in the origin of this asexual group and has played a role in shaping the patterns of genetic diversity observed. This study suggests that genetic signatures of ancient asexuality must be taken with caution due to the confounding effect of interspecific hybridization, which has long been implicated in the origins of apomictic species.


Solid state and solution structure of Boc-L-Ala-delta Phe-delta Phe-NHMe: a dehydropeptide showing propensity for 3(10)-helices of both screw senses.
The crystal and molecular structure of the peptide Boc-L-Ala-delta Phe-delta Phe-NHMe, containing two consecutive dehydro-phenylalanine (delta Phe) residues, has been solved by x-ray diffraction. Two independent molecules, X and Y, are present in the crystallographic unit. Their conformation corresponds approximately to an incipient 3(10)-helix stabilized by two intramolecular hydrogen bonds. The (phi, psi) torsion angles, however, have negative and positive signs in the two molecules X and Y, respectively. Therefore, in spite of the presence of an amino acid residue of the L configuration, the two helical molecules have opposite screw senses, even though the right-handed helix is less distorted than the left-handed one in correspondence of the L-Ala residue. The CD spectra in various solvents exhibit exciton bands originating from dipole-dipole interaction between the delta Phe side chains. Addition of DMSO to the chloroform solution produces, as a first step, a strong increasing of the CD bands, which are then progressively canceled by increasing DMSO concentration. The nmr data parallel the behavior observed in the CD spectra. In CDCl3 solution, the temperature coefficients of the NH resonances are consistent with the involvement of the last two amide protons of the sequence in intramolecular hydrogen bonds, but only negligibly small nuclear Overhauser effects (NOE) are observed. Addition of 5% DMSO-d6 allows the observation of diagnostic NOEs. CD and nmr data indicate that the solid state structure is retained in solution, and are consistent with the presence of right-handed and left-handed conformers, with a prevalence of the more stable right-handed one.


Physiological and psychological effects of alpha and EMG feedback training with college drug users: a pilot study.
Twenty-four volunteer college students who were regular drug users were randomly allocated to three training groups of equal size: alpha feedback, EMG feedback and a joked control group. The subjects were unaware of which feedback condition they received and were asked to practise at home during a six-month follow-up period in order to achieve a relaxed state similar to that experienced during training. No group was successful in retaining gains made in their alpha levels during each session, but the EMG group significantly reduced their muscular activity during training and retained the improvement during follow-up. The alpha and joked groups did not greatly improve their EMG during training but at follow-up achieved the same levels as the EMG group. There was evidence to suggest that a reduction in drug use among light and medium users was maintained during follow-up. Significant and lasting improvements were made by each group in the duration and quality of their sleep and anxiety levels were reduced.


The Mechanisms of Sevoflurane-Induced Neuroinflammation.
Sevoflurane is one of the most commonly used inhaled anesthetics due to its low blood gas coefficient, fast onset, low airway irritation, and aromatic smell. However, recent studies have reported that sevoflurane exposure may have deleterious effects on cognitive function. Although neuroinflammation was most widely mentioned among the established mechanisms of sevoflurane-induced cognitive dysfunction, its upstream mechanisms have yet to be illustrated. Thus, we reviewed the relevant literature and discussed the most mentioned mechanisms, including the modulation of the microglial function, blood-brain barrier (BBB) breakdown, changes in gut microbiota, and ease of cholinergic neurotransmission to help us understand the properties of sevoflurane, providing us new perspectives for the prevention of sevoflurane-induced cognitive impairment.


Skepticism, statistical methods, and the cigarette: a historical analysis of a methodological debate.
The discipline of modern "risk factor" epidemiology was in its formative stages in the early 1950s, when epidemiologic studies revealed a strong association between cigarette smoking and lung cancer mortality. Many medical scientists and physicians were reluctant to accept these studies as a demonstration of causation because the methods were "statistical" and involved data collected in uncontrolled conditions outside the laboratory. But a substantial number of senior biostatisticians and epidemiologists also voiced concerns, albeit more methodologically sophisticated, about the quality of the evidence at the time. Statistical methods were just beginning to work their way into medicine and public health, and many epidemiologists and statisticians were concerned about the potential misuse of these methods by untrained investigators. When studies of smoking and lung cancer gained increasing publicity and were being used to recommend public health policies, some prominent epidemiologists and statisticians highlighted this debate in their efforts to pursue methodological reform. Participants in the debate over smoking and lung cancer saw the need for explicit and rigorous standards for evaluating etiologic hypotheses, but they held conflicting views about what those standards should be. These diverging views reflect an underlying tension within the discipline of epidemiology between the search for "objective" methods of scientific inference and the practical needs of public health research that persists today.


Carotid Body Tumor: Clinical Features.
Although carotid body tumor (CBT) is a rare neoplasm, it should always be considered in differential diagnosis of lateral neck masses. We shared the 10 years of CBT experience in our clinic and started a discussion on CBT with literature support. A total of 21 patients with CBT diagnosis in Fırat University Hospital, Otorhinolaryngology Clinic, participated in the study. Patients were evaluated based on demographical data and particularities of the tumor. Participant patients were 19 women and 2 men, and their ages were between 21 and 79 (mean age 54.06 ± 12.48). The most frequent reason for the patients to apply to the hospital was painless swelling in the neck (76.2%). Tumor was located in the right side of the neck in 10 patients (47.6%), and in the left side of the neck in 11 (52.4%). Twenty patients (95.2%) had undergone computerized tomography angiography. Surgical treatment was applied to 19 patients (90.5%) and the tumor was totally excised. According to Shamblin classification, 15 of the tumors of these patients were class II (78.9%) and 4 were class III (21.1%). In 1 patient (5.3%), postoperative contusion infection that recovered after medical treatment was observed; in 2 patients (10.5%), n. vagus injury was observed because of tumor's pervasion of n. vagus; and in 1 of these patients vocal cord paralysis was developed and this patient was later taken into thyroplasty surgery. Two patients (10.5%) suffered n. hypoglossus injury, 1 of these recovered within 3 months postoperative and the other developed n. hypoglossus palsy. The size and extension of the tumor should be determined by preoperative imaging for the correct planning of surgical procedure. It should be taken into consideration that despite advanced surgical techniques, the rate of postoperative cranial nerve damage is still high.


Actions required to implement integrated care for older people in the community using the World Health Organization's ICOPE approach: A global Delphi consensus study.
Integrated care is recognised as an important enabler to healthy ageing, yet few countries have managed to sustainably deliver integrated care for older people. We aimed to gather global consensus on the key actions required to realign health and long-term systems and integrate services to implement the World Health Organization (WHO) Integrated Care for Older People (ICOPE) approach. A two-round eDelphi study, including a global consultation meeting, was undertaken to identify, refine and generate consensus on the actions required across high-, middle- and low-income countries to implement the WHO ICOPE approach. In round 1, a framework of 31 actions, empirically derived from previous WHO evidence reviews was presented to panellists to judge the relative importance of each action (numeric rating scale; range:1-9) and provide free-text comments concerning the scope of the actions. These outcomes were discussed and debated at the global consultation meeting. In round 2, a revised framework of 19 actions was presented to panellists to measure their extent of agreement and identify 'essential' actions (five-point Likert scale; range: strongly agree to strongly disagree). A threshold of ≥80% for agree/strongly agree was set a priori for consensus. After round 1 (n = 80 panellists), median scores across 31 actions ranged from 6 to 9. Based on pre-defined category thresholds for median scores, panellists considered 28 actions (90·3%) as 'important' and three (9·7%) as 'uncertain'. Fifteen additional actions were suggested for inclusion based on free-text comments, creating 46 for consideration at the global consultation meeting. In round 2 (n = 84 panellists), agreement (agree or strongly agree) ranged from 84·6-97·6%, suggesting consensus. Fourteen (73·7%) actions were rated as essential. Fourteen essential actions and five important actions are necessary at system (macro; n = 10) and service (meso; n = 9) levels to implement community-based integrated care for older people.


The genetic determination of plasma apolipoprotein A-I levels measured by radioimmunoassay: a study of high-risk pedigrees.
Apolipoprotein A-I (apo A-I) is the major protein component of the high-density lipoprotein (HDL) found in all primates. Using radioimmunoassay, we measured plasma apo A-I levels in 97 individuals from 23 pedigrees ascertained through cases of hypertension or early coronary artery disease (CAD). Using complex segregation analysis, we found that a genetic model with both a single locus with a major effect and polygenic loci gave the best explanation for the distribution of apo A-I levels in these pedigrees. There was no evidence for a major locus effect on HDL cholesterol in these pedigrees. This is the first study to show evidence of a major effect of a single genetic locus on the quantitative variation of plasma apo A-I in a sample of pedigrees enriched for individuals at risk for CAD.


Modified local sands for the mitigation of harmful algal blooms.
A new method was developed for marine harmful algal bloom (HAB) mitigation using local beach sand or silica sand modified with chitosan and polyaluminum chloride (PAC). Untreated sand was ineffective in flocculating algal cells, but 80% removal efficiency was achieved for Amphidinium carterae Hulburt and a Chlorella sp. in 3 min (t80 = 3 min) using 120 mg L-1 sand modified with 10 mg L-1 PAC and 10 mg L-1 chitosan. After several hours 92% - 96% removal was achieved. The t80 for removing A. carterae using the modifiers only (PAC and chitosan combined) was 60 min and for Chlorella sp. 120 min, times which are much slower than with the corresponding modified sand. Sands were critical for speeding up the kinetic processes of flocculation and sedimentation of algal flocs. PAC was helpful in forming small flocs and chitosan is essential to bridge the small flocs into large dense flocs. Chitosan was also important in inhibiting the escape of cells from the flocs. Chitosan and PAC used together as modifiers make it possible to use local beach sands for HAB mitigation in seawater. Economical and environmental concerns could be reduced through the use of sands and biodegradable chitosan, but the potential impacts of PAC need further study.


Structural features of membrane fusion between influenza virus and liposome as revealed by quick-freezing electron microscopy.
The structure of membrane fusion intermediates between the A/PR/8(H1N1) strain of influenza virus and a liposome composed of egg phosphatidylcholine, cholesterol, and glycophorin was studied using quick-freezing electron microscopy. Fusion by viral hemagglutinin protein was induced at pH 5.0 and 23 degrees C. After a 19-s incubation under these conditions, small protrusions with a diameter of 10-20 nm were found on the fractured convex faces of the liposomal membranes, and small pits complementary to the protrusions were found on the concave faces. The protrusions and pits corresponded to fractured parts of outward bendings of the lipid bilayer or "microprotrusions of the lipid bilayer." At the loci of the protrusions and pits, liposomal membranes had local contacts with viral membranes. In many cases both the protrusions and the pits were aligned in regular polygonal arrangements, which were thought to reflect the array of hemagglutinin spikes on the viral surface. These structures were induced only when the medium was acidic with the virus present. Based on these observations, it was concluded that the microprotrusions of the lipid bilayer are induced by hemagglutinin protein. Furthermore, morphological evidence for the formation of the "initial fusion pore" at the microprotrusion was obtained. The protrusion on the convex face sometimes had a tiny hole with a diameter of <4 nm in the center. The pits transformed into narrow membrane connections <10 nm in width, bridging viruses and liposomes. The structures of the fusion pore and fusion neck with larger sizes were also observed, indicating growth of the protrusions and pits to distinct fusion sites. We propose that the microprotrusion of the lipid bilayer is a fusion intermediate induced by hemagglutinin protein, and suggest that the extraordinarily high curvature of this membrane structure is a clue to the onset of fusion. The possible architecture of the fusion intermediate is discussed with regard to the localization of intramembrane particles at the microprotrusion.


[A rare case of a post-traumatic intraventricular defect].
Cardiac injury occurs in about 20-30% of major chest traumas; 80% of penetrating cardiac wounds are fatal; the survival rate is related to the mechanism of injury (whether by stab wound or gunshot wound), the extent of injury, the need for emergency room thoracotomy for resuscitation, and the presence of cardiac tamponade. The right ventricle and atrium are the most commonly involved cardiac chambers. We present a case of 63-year-old male patient who presented with a systolic murmur after a chest wall stab wound. Transthoracic echocardiogram revealed a small pericardial effusion and laceration of the interventricular septum. Cardiac catheterization revealed a moderate increase in mean pulmonary artery pressure and a moderate left to right shunt, due to interventricular defect which was closed percutaneously. The presented case is very unusual: the clinical picture was very poor because of the spontaneous closure of the wound, the absence of signs of cardiac tamponade, and the moderate size of the defect. We emphasize the role of echocardiography in the anatomic and functional evaluation of such lesions.


Demographic features, laboratory findings and outcomes of ICU patients with COVID-19 in Ahvaz, Iran: A single-centered prospective study.
COVID-19 can lead to severe acute respiratory syndrome so that some patients need to be admitted to the Intensive Care Unit (ICU). The aim of the current study is to investigate the frequency of demographic, laboratory and imaging findings and type of treatment and their relationship with disease outcomes in patients with COVID-19. This prospective cross-sectional study was conducted on all patients with COVID-19 who were admitted in the ICU of Razi Hospital in Ahvaz, Iran from January 20 to February 20, 2021. Patient information including demographic features, laboratory and imaging findings and clinical outcomes was recorded. One hundred and thirty-three patients were recruited in the present study, out of which 74 patients (55.6%) were males and 59 patients (44.4%) were females. The overall mortality rate of patients was 35.3% (47 patients) and was higher in patients over 65 years of age. There was a significant difference in terms of thrombocytopenia (P value: 0.001), lymphopenia (P value: 0.004), progression of lung involvement in imaging, shock, disseminated intravascular coagulation (DIC), sepsis and receiving invasive respiratory support in living and deceased patients (P value < 0.001). Furthermore, the difference in life status and the length of in-ICU stay in patients with hyperkalemia and renal failure was statistically significant (P value = 0.033, P value < 0.001 respectively). Mortality rate of patients with COVID- 19 admitted to ICU is generally high. According to the findings of this study, thrombocytopenia, lymphopenia, hyperkalemia and AKI are laboratory disorders associated with increased mortality. Moreover, the progression of pulmonary involvement in imaging, shock, DIC, sepsis, and need to invasive respiratory support is associated with low survival of patients.


Evaluation of in vitro activity of ceftolozane-tazobactam against recent clinical bacterial isolates from Brazil - the EM200 study.
The emergence of antibiotic resistance is increasing and there are few effective antibiotics to treat infections caused by resistant and multidrug resistant bacterial pathogens. This study aimed to evaluate the in vitro activity of ceftolozane-tazobactam against clinical bacterial isolates from Brazil. A total of 673 Gram-negative bacterial isolates including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and other Enterobacterales collected from 2016 to 2017 were tested, most of them isolated from patients in intensive care units. Minimum inhibitory concentrations (MIC50/90) were determined by broth microdilution for amikacin, aztreonam, cefepime, cefotaxime, cefoxitin, ceftolozane-tazobactam, ceftazidime, ceftriaxone, ciprofloxacin, colistin, ertapenem, imipenem, levofloxacin, meropenem, and piperacillin-tazobactam using dried panels. Antimicrobial susceptibility results were interpreted according to Clinical and Laboratory Standards Institute criteria. Susceptibility rates to ceftolozane-tazobactam ranged from 40.4% to 94.9%. P. aeruginosa susceptibility rate to ceftolozane-tazobactam was 84.9% (MIC50/90, 1/16μg/mL) and 99.2% to colistin. For E. coli, ceftolozane-tazobactam inhibited 94.9% (MIC50/90, 0.25/1μg/mL) of the microorganisms. The susceptibility rate of K. pneumoniae to ceftolozane-tazobactam was 40.4% (MIC50/90, 16/>32μg/mL). Other Enterobacterales have shown susceptibility rates of 81.1% (MIC50/90, 0.5/16μg/mL) to ceftolozane-tazobactam, 93.9% to meropenem, 90.9% to amikacin (90.9%), and 88.6% to ertapenem. In non-carbapenemase producing isolates, AmpC mutations were found three isolates. Ceftolozane-tazobactam has shown relevant activity against a large variety of the analyzed microorganisms collected from multiple centers in Brazil, showing promising results even in multidrug resistant strains.


Analgesia in esophagectomy: a narrative review.
Optimal pain management for esophagectomy facilitates prevention of postoperative complications such as pneumonia, but also chronic pain. Historically, multimodal intravenous analgesia was employed. In the last decades, regional anesthesia including epidural and paravertebral analgesia is frequently used. In this narrative review, we provide a comprehensive overview of the available evidence for the different analgesia regimens for esophagectomy. A search was conducted in the PubMed/MEDLINE database in November 2022. Only reports in English or Dutch were included. Editorials or articles lacking full text were excluded. A review of different analgesia regimens after esophagectomy is provided. Epidural analgesia (EA) was suggested to reduce postoperative pneumonia and prevent chronic postsurgical pain (CPSP) as compared to opioid-based systemic analgesia and was considered the gold standard of pain management for esophagectomy. In the last decades, the side-effects of EA became more evident. Next to mild or moderate side-effects such as hypotension and urinary retention, several reports emphasized the incidence of serious neurologic complications to be much higher than estimated before. In addition, minimally invasive surgery fostered that other regional analgesia (RA) techniques are potential alternatives for EA. Paravertebral catheter placement can be performed under videoscope view during the thoracic phase of esophagectomy, making it a safe and easily placed block. Evidence on the effectiveness of erector spinae plane block (ESPB) is limited in this context. Several analgesia regimens after esophagectomy are described. EA is most common, however paravertebral analgesia is a good alternative. Other techniques are also gaining ground but randomized clinical trials are lacking. Future studies should focus on the efficacy of paravertebral and erector spinae blocks for postoperative pain management for esophagectomy.


Studies dealing with hepatic RNA metabolism in rats force-fed a threonine-devoid diet.
Young rats were force-fed for 3 days a purified diet devoid of threonine and a number of aspects relating to RNA metabolism in the livers were studied. The findings in the livers of rats force-fed the threonine-devoid diet in comparison with those force-fed the complete diet were as follows: a) poly(A)-mRNA was increased in nuclei and in polyribosomes; b) DNA-dependent RNA polymerases I and II activities were increased; c) in vitro release of 14C-orotic acid labeled RNA from nuclei revealed that transport was unchanged and nucleoside triphosphatase activity of nuclear envelopes was unchanged; d) polyribosomes (total, free and membrane-bound) shifted toward heavier aggregation and in vitro 14C-leucine incorporation into protein was increased; e) RNase activities (at pH 5.4, 7.6, 9.5) were essentially unaltered; and f) in vivo 14C-choline incorporation into microsomal membranes was increased. By administering selected inhibitors of RNA and protein synthesis, such as actinomycin D, alpha-amanitin or cycloheximide, prior to killing the rats force-fed the threonine-devoid or complete diet for 3 days, it was demonstrated that the stimulatory effect on hepatic polyribosomes and protein synthesis in the experimental group was dependent upon new synthesis of poly(A)mRNA and of protein.


Bell's palsy in Singapore: a view from the patient's perspective.
Bell's palsy is a well-recognised disease with robust research on its possible aetiologies and epidemiology, but scant information on patients' concerns and concepts regarding the condition is available. We aimed to evaluate the ideas, concerns and expectations of patients with Bell's palsy in Singapore. A cross-sectional study was conducted at a single tertiary-care hospital in Singapore. Participants were all patients with newly diagnosed Bell's palsy referred to the otolaryngology department either from the emergency department or by general practitioners. Participants were given a self-administered questionnaire and their facial nerve palsies were graded by the consultant doctor. A total of 52 patients were recruited, of which 41 were available for analysis. 78.0% of patients were concerned that they were having a stroke upon presentation of the symptoms. Other beliefs about the cause of the disease included overwork or stress (36.6%), something that the patient had eaten (9.8%) and supernatural forces (2.4%). About 50% of patients had tried some form of complementary or alternative therapy other than the steroids/medicines prescribed by their general practitioner or emergency physician. While 39.0% of patients agreed that the Internet had helped them understand more about their condition in addition to the information provided by the physician, 9.8% of them specifically disagreed with this statement. We have found that patients with Bell's palsy in Singapore are not very knowledgeable about the disease. Although the Internet is a useful resource, a physician's explanation of the disease and its natural progression remains of utmost importance.


Effects of maxillary expansion on the upper airways: Evidence from systematic reviews and meta-analyses.
Constricted maxilla is frequently associated with reduced nasal airway dimensions. Wheatear skeletal maxillary expansion (ME) is effective on the dimension of the upper airways is still a debated issue. This overview aimed to report the evidence provided by systematic reviews (SRs) on the effect of ME on the upper airways and to assess the methodological quality of the included SRs. Six electronic databases have been explored up to November 2017. After title and abstract screening, SRs addressing the effects of fixed palatal expanders on the dimension and function of the nasal airways were included. The methodological quality of the included SRs was assessed using the updated version of A Measurement Tool to Assess Systematic Review (AMSTAR-2). Eight SRs were included. The methodological quality of most of the included SRs ranged between low and critically low. One SR was rated of high quality. A significant increase in nasal linear dimensions was reported both in the short and long term, but supported by low-/critically low-quality SRs. The significant increase in nasal cavity volume was the only outcome supported by a high-quality SR Controversial results were found with regards to nasal function. Whenever a constricted maxilla is present general dentists, paediatricians and ENTs should be familiar with the potential improvement provided by ME. However, due to the low/critically low quality of SRs supporting these results, ME cannot be indicated only for upper airways enhancement, but should be supported by an orthodontic indication.


Mucin-type O-glycosylation in Mesocestoides vogae (syn. corti).
Protein glycosylation is an important post-translational modification underlying host-parasite interactions, which may determine the outcome of infection. Although Mesocestoides vogae represents an important model for investigating the various aspects of cestode biology, virtually no information is available about the structure and synthesis of glycans in this parasite. In this work, focused on the initiation pathway of mucin-type O-glycosylation in M. vogae, we characterized O-glycoproteins bearing the simple mucin-type cancer-associated Tn and sialyl-Tn antigens, and the expression and activity of ppGalNAc-T, the key enzyme responsible for the first step of mucin-type O-glycosylation. Using immunohistochemistry, Tn and sialyl-Tn antigens were detected mainly in the tegument (microtriches) and in parenchymal cells. Tn expression was also observed in lateral nerve cords. Both Tn and sialyl-Tn antigens were detected in in vitro cultured parasites. Based on their electrophoretic mobility, Tn- and sialyl-Tn-bearing glycoproteins from M. vogae were separated into several components of 22 to 60 kDa. The observation that Tn and sialyl-Tn glycoproteins remained in the 0.6N perchloric acid-soluble fraction suggested that they could be good candidates for characterizing mucin-type glycosylation in this parasite. O-glycoproteins were purified and initially characterized using a proteomic approach. Immunohistochemical analysis of the tissue distribution of ppGalNAc-T revealed that this enzyme is expressed in the sub-tegumental region and in the parenchyma of the parasite. In M. vogae cultured in vitro, ppGalNAc-T was mainly detected in the suckers. Using a panel of 8 acceptor substrate synthetic peptides, we found that M. vogae ppGalNAc-T preferentially glycosylate threonine residues, the best substrates being peptides derived from human mucin MUC1 and from Trypanosoma cruzi mucin. These results suggest that M. vogae might represent a useful model to study O-glycosylation, and provide new research avenues for future studies on the glycopathobiology of helminth parasites.


Development of an indirect ELISA for the detection of serum antibodies to Aspergillus fumigatus in captive penguins.
Aspergillosis is a significant cause of mortality in captive penguins (Sphenisciformes). An indirect ELISA for the detection of Aspergillus fumigatus-specific immunoglobulin in penguins was developed and standardised by making use of a family-specific antiserum (anti-Aptenodyptes patagonica patagonicus). The results were calculated quantitatively as ELISA units, derived by polynomial regression analysis, and semi-quantitatively as end titres. Serum samples from 61 captive penguins were tested with the assay, and the results were compared with those obtained by counterimmunoelectrophoresis (CIE). The ELISA results correlated with the CIE results only when end titres were reported (R(s) = -0.676, P < 0.002). Fifty-seven of the penguins (93 per cent) were seropositive, but the detection of immunoglobulin did not correlate with clinical disease. At Whipsnade Wild Animal Park, Humboldt's penguins (Spheniscus humboldti) demonstrated higher seropositivity than king penguins (Aptenodyptes patagonicapatagonicus) (P = 0.022), but Humboldt's penguins at Fota Wildlife Park had a significantly higher seropositivity than Humboldt's penguins at Whipsnade (P = 0.035).


Psychosocial characteristics of women and men attending infertility counselling.
Little is known about the psychosocial characteristics of infertile couples seeking psychological help. This study describes couples attending infertility counselling. Questionnaires pertaining to socio-demographic factors, motives for wanting a child, lay aetiology of their infertility, dimensions of life and partnership satisfaction, and a complaints list were completed by 974 women and 906 men. Of those who indicated an openness to counselling, almost half actually attended infertility counselling, and two groups, 'no counselling' (358 women and 292 male partners) and 'taking up counselling' (275 women and 243 male partners), were therefore compared. More couples with stressful life events were found in the counselling group. For women taking up counselling, psychological distress, in the form of suffering from childlessness and depression as well as subjective excessive demand (as a potential cause for infertility), was higher in comparison to women not counselled. The higher distress for men in the counselling group was indicated by relative dissatisfaction with partnership and sexuality and by accentuating the women's depression. Infertile couples seeking psychological help are characterized by high levels of psychological distress, primarily in women. The women's distress seems to be more important for attending infertility counselling than that of the men.


ESTIMATING RELATEDNESS USING GENETIC MARKERS.
A new method is described for estimating genetic relatedness from genetic markers such as protein polymorphisms. It is based on Grafen's (1985) relatedness coefficient and is most easily interpreted in terms of identity by descent rather than as a genetic regression. It has several advantages over methods currently in use: it eliminates a downward bias for small sample sizes; it improves estimation of relatedness for subsets of population samples; and it allows estimation of relatedness for a single group or for a single pair of individuals. Individual estimates of relatedness tend to be highly variable but, in aggregate, can still be very useful as data for nonparametric tests. Such tests allow testing for differences in relatedness between two samples or for correlating individual relatedness values with another variable.


Ipsilateral but not contralateral blockade of excitatory amino acid receptors in the caudal ventrolateral medulla inhibits aortic baroreceptor reflex in rats.
The caudal ventrolateral medulla (CVLM) contains vasodepressor neurons which, when activated, decrease vasomotor tone. To investigate whether excitatory amino acid receptors in the CVLM of the rat are involved in mediation of the aortic baroreceptor reflex, we microinjected amino acid antagonists unilaterally into the CVLM and examined their effects on the depressor response to electrical stimulation of the aortic nerve which contains mainly baroreceptor afferent fibers in rats. Male Wistar rats were anaesthetized with urethane, paralyzed and artificially ventilated. To block reflex vagal effects, methylatropine (1 mg/kg) was given intravenously. Kynurenate (227 ng), an excitatory amino acid antagonist, injected ipsilaterally but not contralaterally into the CVLM markedly inhibited the depressor response to aortic nerve stimulation, while both injections produced a similar small increase in basal blood pressure. Muscimol (1 ng), a GABA receptor agonist, injected ipsilaterally into the CVLM partly inhibited the baroreflex response, while it produced a moderate increase in basal blood pressure. 2-Amino-5-phosphonovalerate (APV) (10 ng), a N-methyl-D-aspartate (NMDA) receptor antagonist, and MK-801 (30 ng), a NMDA receptor channel blocker, partly inhibited the baroreflex response. MK-801 (30 ng) injected into the CVLM reduced the depressor response to the NMDA receptor agonist NMDA (0.3 ng) but not to the quisqualate receptor agonist quisqualate (0.1 ng) and the kainate receptor agonist kainate (0.1 ng), while kynurenate (227 ng) inhibited the depressor response to all three excitatory amino acid receptor agonists. These findings provide further evidence for the presence of excitatory amino acid receptors involved in mediating the aortic baroreceptor reflex in the rat CVLM.(ABSTRACT TRUNCATED AT 250 WORDS)


Sex-specific behavioral outcomes of early-life adversity and emerging microglia-dependent mechanisms.
Early-life adversity (ELA) is known to alter brain circuit maturation as well as increase vulnerability to cognitive and emotional disorders. However, the importance of examining sex as a biological variable when researching the effects of ELA has not been considered until recently. This perspective discusses the sex-specific behavioral outcomes of ELA in both humans and animal models, then proposes microglia-mediated mechanisms as a potential underlying cause. Recent work in rodent models suggests that ELA provokes cognitive deficits, anhedonia, and alcohol abuse primarily in males, whereas females exhibit greater risk-taking and opioid addiction-related behaviors. In addition, emerging evidence identifies microglia as a key target of ELA. For example, we have recently shown that ELA inhibits microglial synapse engulfment and process dynamics in male mice, leading to an increase in excitatory synapse number onto corticotrophin-releasing hormone (CRH)-expressing neurons in the paraventricular nucleus of the hypothalamus (PVN) and aberrant stress responses later in life. However, ELA-induced synaptic rewiring of neural circuits differs in females during development, resulting in divergent behavioral outcomes. Thus, examining the role of microglia in the sex-specific mechanisms underlying ELA-induced neuropsychiatric disorders is an important topic for future research.


How can the study of biological processes help design new interventions for children with severe antisocial behavior?
Children with severe antisocial behavior have an increased risk of showing violently aggressive and other forms of problem behavior in adolescence and adulthood. It is well established that both biological and social factors are involved in the development of antisocial behavior. The primary aim of this paper is to discuss the evidence that specific neurobiological systems are involved in the etiology of childhood-onset antisocial behavior. These factors are responsible for the severity of the behavioral problems observed in antisocial children, but they also play a role in their persistence, because they influence children's interactions with their environment. We will discuss the possible causes of disruptions in neurobiological systems in childhood antisocial behavior and point out the implications of these findings for theory and clinical practice. We will argue that familial factors (e.g., genetic influences, early childhood adversity) are linked to negative behavioral outcomes (e.g., antisocial behavior problems) through the mediating and transactional interplay with neurobiological deficits. An investigation of neurobiological functioning in antisocial children might not only indicate which children are most likely to persist in engaging in severe antisocial behavior, but also guide the development of new interventions.