Using Games to Promote English as a Foreign Language Learners' Willingness to Communicate: Potential Effects and Teachers' Attitude in Focus.
This study aims to find out the role of games in promoting students' willingness to communicate (WTC) and their teachers' attitude toward it. In order to collect the data, the researchers employed a 28-item questionnaire which was given to 60 English as a Foreign Language (EFL) learners in an English institute. Then, the students were randomly divided into two groups of 30 learners functioning as control and experimental groups. The students in the experimental group received games in their language lessons and classes, while control group learners did not. At the end of the term, the same questionnaire was given to the students to know if playing games had a significant impact on their WTC. In addition, the teachers were asked to answer a 30-item questionnaire to investigate their attitudes toward playing games in language classes. The results showed that most of the teachers in this study believe that games have a positive influence on the students' attitudes towards learning English and that using them in class serves many educational purposes. In addition, games played a significant role in improving the EFL leaners' willingness to communicate. In the light of these findings, the researchers suggested using games as energizers and practical activities at the end of class not only to improve enthusiasm for learning, but also to improve the learners' WTC.


Improved graft function in liver-transplanted patients after partial splenic embolization: reversal of splenic artery steal syndrome?
To describe the functional effect of partial splenic embolization (PSE) in liver-transplanted (LT) patients with hypersplenism and hepatitis C virus (HCV) recurrence. From May 2002 to May 2005, five LT patients with persistent hypersplenism, viral recurrence and graft dysfunction underwent PSE prior to pegylated interferon/ribavirin (peg-IFN/RBV). The mean splenic size was 19.5 cm (16-21) and there was evidence of an enlarged splenic artery (10.7+/-1 mm). PSE produced a median splenic infarction of 90% and a significant reduction in the splenic artery diameter to 5.8+/-0.4 mm (p=0.04). PSE significantly improved hematologic parameters, bilirubin levels, prothrombin activity, international normalized ratio and the Model for End-stage Liver Disease (MELD) score despite high HCV-RNA (6.2 log10 IU/mL). It was demonstrated histologic amelioration of ischemic changes in all subjects. PSE allowed the safe use of full-dose peg-IFN plus RBV in all subjects. In HCV LT patients with chronic graft dysfunction and cholestasis the improvement in the liver function following PSE might be due to the reversal of an undiagnosed splenic artery steal syndrome related to chronic hypersplenism masked by HCV recurrence.


Esophageal bronchus in an infant--a rare cause of recurrent pneumonia.
A 4-month-old female infant presented to our institute with recurrent pneumonia and was diagnosed with a right lower lobe esophageal bronchus. This congenital anomaly is extremely rare with very few cases reported in the literature. We describe the diagnostic workup and management that led to a successful outcome.


Successful immunization of naturally reared pigs against porcine cysticercosis with a recombinant oncosphere antigen vaccine.
Taenia solium causes cysticercosis in pigs and taeniasis and neurocysticercosis in humans. Oncosphere antigens have proven to be effective as vaccines to protect pigs against an experimental infection with T. solium. A pair-matched vaccination trial field, using a combination of two recombinant antigens, TSOL16 and TSOL18, was undertaken in rural villages of Peru to evaluate the efficacy of this vaccine under natural conditions. Pairs of pigs (n=137) comprising one vaccinated and one control animal, were allocated to local villagers. Animals received two vaccinations with 200 μg of each of TSOL16 and TSOL18, plus 5mg Quil-A. Necropsies were performed 7 months after the animals were distributed to the farmers. Vaccination reduced 99.7% and 99.9% (p<0.01) the total number of cysts and the number of viable cysts, respectively. Immunization with the TSOL16-TSOL18 vaccines has the potential to control T. solium transmission in areas where the disease is endemic, reducing the source for tapeworm infections in humans.


The use of nanomechanic sensor for studies of morphofunctional properties of lymphocytes from healthy donors and patients with chronic lymphoblastic leukemia.
Tapping mode-atomic force microscopy and force spectroscopy were used for studies of the topography of the cell surface and elastic properties of lymphocytes from healthy donors and patients with chronic lymphoblastic leukemia. It was demonstrated that the decrease in lymphocytes stiffness in patients with chronic lymphoblastic leukemia by 51.4% (p<0.05) was accompanied by spatial modification of the cell surface, in particular, increase in the number of globular protrusions and depressions by 247 and 122%, respectively (p<0.05), in comparison with normal lymphocytes.


25-Hydroxyvitamin D [25(OH)D] levels and diabetic foot ulcer: is there any relationship?
In recent years, there has been an effort to understand possible roles of 25(OH)D, including its role in the immune system particularly on T cell medicated immunity, pancreatic insulin secretion and insulin action. 25(OH)D stimulates the cell differentiation and reduces cell proliferation, which is essential for cell growth and wound healing. However, data on the association between low level of plasma 25(OH)D and diabetic foot syndrome are scarce. Circulating plasma levels of 25(OH)D were measured in diabetic patients with ulcer (n=162) and without ulcer (n=162) in a prospective cohort hospital based study. Of these patients, 85.1% had type 2 diabetes. Subjects with diabetic foot ulcer showed lower median plasma level of 25(OH)D [6.3(4.2-11.1) vs 28.0(21.4-37.0)] ng/ml after adjusting the age and BMI. Regardless of the low levels of 25(OH)D in cases and controls, it was associated with neuropathy, sex (female), duration of ulcer healing, and smoking status and independent of confounding factors, including BMI (kg/m²), A1c (%), hypertension, nephropathy, foot ulcer, retinopathy, CAD, PAD, HDL-C (mg/dl) and LDL-C (mg/dl). The factors which predict the risk of developing ulcer independent of 25(OH)D status were A1c (>6.9%) [OR 4.37; RR 1.77], HDL-C (<40mg/dl) [OR 1.16; RR 1.07], LDL-C (>100mg/dl) [OR 1.07; RR 1.03], triglycerides (>200mg/dl) [OR 1.40; RR 1.19], neuropathy [OR 6.88; RR 3.12], retinopathy [OR 3.34; RR 1.91], hypertension [OR 1.64; RR 1.28], nephropathy [OR 3.12; RR 1.87] and smoking [OR 4.53; RR 2.99] using odds and risk ratios. It is not clear whether the suppression of delayed wound healing seen during 25(OH)D deficiency is due to the secondary effect or is a direct action of vitamin D on certain components of the immune system. Long-term randomized trials are needed to see the impact of vitamin D supplementation on the outcome of diabetic foot patients.


Treatment of tumours of the posterior part of the third ventricle and the pineal region: a long term follow-up.
This paper represents a further investigation into the origninal series, with additional cases from N.I.N.U. This follow-up has been immeasureably enhanced by the availability of the EMI Scanner. It seems clear that there continues to be adequate evidence that the use of shunting procedures, followed by radiotherapy, is a rational approach to tumours of the pineal and posterior third ventricular region. Short of more extensive evidence to the contrary, we feel that it remains the method of choice in the initial treatment of these difficult lesions.


Need for improved detection of voluntary medical male circumcision adverse events in Mozambique: a mixed-methods assessment.
Adverse events (AE) resulting from voluntary medical male circumcision (VMMC) are commonly used to measure program quality. Mozambique's VMMC program data reports a combined moderate and severe AE rate of 0.2% through passive surveillance. With active surveillance, similar programs report AE rates ranging from 1.0 to 17.0%. The objective of this activity was to assess potential underreporting of AEs via the passive surveillance system in Mozambique. This mixed-methods assessment randomly selected one third (16) of all 46 VMMC clinics through stratified sampling, based on volume. A retrospective record review was conducted including patient clinical files, stock records of Amoxicillin/Clavulanic Acid (the choice antibiotic for VMMC-related infections), and clinic-level AE rates from the national database. Records from the month of April 21 to May 20, 2017 were analyzed to identify both reported and potentially unreported AEs. In addition, external, expert clinicians observed post-operative visits (n = 167). Descriptive statistics were calculated, including difference between reported and identified AEs, an adjusted retrospective AE rate, and an observed prospective AE rate in each clinic. A total of 5352 circumcisions were performed in the 16 clinics: 8 (0.15%) AEs were reported. Retrospective clinical record reviews identified 36 AEs (0.67%); AE severity or type was unknown. Using Amoxicillin/Clavulanic Acid dispensation as a proxy for VMMC-related infections, 39 additional AEs infections were identified, resulting in an adjusted AE rate of 1.4%, an 8.3 fold increase from the reported AE rate. Prospective, post-operative visit observations of 167 clients found 10 AEs (5.9%); infection was common and boys 10-14 years old represented 80% of AE clients. Evidence suggests underreporting of AEs in the Mozambican VMMC program. Quality improvement efforts should be implemented in all VMMC sites to improve AE identification, documentation and prevention efforts.


Preparation of surfaces of composite samples for tip based micro-analyses using ion beam milling.
Ion beam milling, as a method of surface design for tip analytical techniques, was explored. A sample of clay, embedded in a resin, was treated by the ion beam and allowed AFM (a typical tip technique) to be successfully applied. The method is suitable for advanced tip analyses based on AFM, like TERS or SNOM, and for samples that are not possible to prepare by standard mechanical methods. The approach can be useful for characterisation of the surfaces of many different types of materials in versatile applications such as catalysis, corrosion science or advanced material characterisation.


The effects of nalmefene on emotion processing in alcohol use disorder - A randomized, controlled fMRI study.
Nalmefene is a µ- and δ-opioid receptor antagonist and a partial κ-opioid receptor agonist. The drug is suggested to reduce the craving for, and the consumption of alcohol effectively, also alleviating anxiety and anhedonia. The present fMRI study is the first to investigate the processing of emotions as a possible mechanism of action of nalmefene in humans. Fifteen non-treatment-seeking participants suffering from alcohol use disorder (AUD) (24-66 years; 5 females) finished this randomized, placebo controlled, double blind study. Following a cross over design, participants received either a single dose nalmefene or a placebo, with an interval of one week between sessions. Using fMRI, we investigated neural reactivity during the presentation of emotional faces picture sets. Additionally, we performed a visual dot-probe task to detect nalmefene's effects on attentional bias. We detected an increase in the response to emotional faces in the supramarginal gyrus, the angular gyrus as well as the putamen in the nalmefene vs. placebo condition. However, contradictory to our initial hypotheses, amygdala activation was not altered significantly in the placebo condition - a limitation, which might be associated with a lack of activation in the placebo condition maybe due to the small sample size. Attentional bias analyses revealed an interaction effect by trend, which was driven by a significant effect in a sub-analysis showing increased attentional shift towards happy compared to fearful facial expressions under nalmefene. Nalmefene increased brain activation in areas responsible for empathy, social cognition and behavior, which might help alleviating the reinforcing properties of alcohol.


Deep inferior epigastric perforator (DIEP) flap safety profile in slim versus non-slim BMI patients: A systematic review and meta-analysis.
The complications of breast reconstruction using deep inferior epigastric perforator (DIEP) flaps in non-slim patients are well recognized. However, the effects of this surgery performed on slim patients are yet to be consolidated. This study aims to compare the outcomes of performing DIEP flap breast reconstruction in slim and non-slim body mass index (BMI) patients. Meta-analysis was performed with a multi-database search (Cochrane, EMBASE, OVID Medline, PubMed, and Web of Sciences) according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines on 1 February 2021. Data from articles meeting inclusion criteria were extracted and analyzed with an inverse variance statistical model. Seven studies were included, comprising 574 slim patients and 901 non-slim BMI patients who underwent autologous DIEP flap breast reconstruction. When comparing between the slim and non-slim groups, no statistically significant difference was found in terms of complete flap loss (OR=0.53, 95% CI: 0.11-2.68, p=0.44), partial flap loss (OR=0.92, 95% CI: 0.3-2.82, p=0.88), and fat necrosis (OR=0.91, 95% CI: 0.61-1.37, p=0.66). Similarly, in terms of general surgical complications, there was no statistically significant difference between groups in terms of all complications (OR=0.83, 95% CI: 0.45-1.51, p=0.54), abdominal wound healing complications (OR=1.01, 95% CI: 0.59-1.73, p=0.97), infections (OR=0.74, 95% CI: 0.41-1.37, p=0.34), and seroma (OR=0.89, 95% CI: 0.35-227, p=0.81). There is no increased risk of postoperative complications in either group. DIEP flaps can be safely performed in slim patients, though higher quality research may be required to further confirm this.


Aspartic acid in the arcuate nucleus attenuates the depressive effects of naloxone on ventilation.
Ventilation, oxygen consumption, the ventilatory equivalent for oxygen, and ventilatory responses to hypoxia and to hypercapnia were evaluated in conscious male rats who received each of four treatments: (1) microinjection of artificial cerebrospinal fluid (aCSF) into the arcuate nucleus and subcutaneously saline (CS); (2) aspartic acid into the arcuate nucleus and saline subcutaneously (AS); (3) aCSF into the arcuate nucleus and naloxone subcutaneously (CN); and (4) aspartic acid into the arcuate nucleus and naloxone subcutaneously (AN). Rats treated with CN exhibited a depression of ventilation, ventilatory equivalent, ventilatory response to hypercapnia, and tidal volume response to hypoxia and to hypercapnia. AS had no effect on any parameters. Administration of both aspartic acid and naloxone attenuated all the effects of CN except the depression of minute ventilation in response to hypercapnia. Therefore the naloxone (a mu opioid receptor antagonist) induced a depression of ventilation that was attenuated by aspartic acid acting on N-methyl-D-aspartic acid receptors in the arcuate nucleus.


Diagnostic Accuracy of the Explicit Diagnostic Criteria for Transient Ischemic Attack: A Validation Study.
Background and Purpose- The clinical diagnosis of a transient ischemic attack (TIA) can be difficult. Evidence-based criteria hardly exist. We evaluated if the recently proposed Explicit Diagnostic Criteria for TIA (EDCT), an easy to perform clinical tool focusing on type, duration, and mode of onset of clinical features, would facilitate the clinical diagnosis of TIA. Methods- We used data from patients suspected of a TIA by a general practitioner and referred to a TIA service in the region of Utrecht, the Netherlands, who participated in the MIND-TIA (Markers in the Diagnosis of TIA) study. Information about the clinical features was collected with a standardized questionnaire within 72 hours after onset. A panel of 3 experienced neurologists ultimately determined the definite diagnosis based on all available diagnostic information including a 6-month follow-up period. Two researchers scored the EDCT. Sensitivity, specificity, and predictive values of the EDCT were assessed using the panel diagnosis as reference. A secondary analysis was performed with modified subcriteria of the EDCT. Results- Of the 206 patients, 126 (61%) had a TIA (n=104) or minor stroke (n=22), and 80 (39%) an alternative diagnosis. Most common alternative diagnoses were migraine with aura (n=24; 30.0%), stress related or somatoform symptoms (n=16; 20.0%), and syncope (n=9; 11.3%). The original EDCT had a sensitivity of 98.4% (95% CI, 94.4-99.8) and a specificity of 61.3% (49.7-71.9). Negative and positive predictive values were 96.1% (86.0-99.0) and 80.0% (75.2-84.1), respectively. The modified EDCT showed a higher specificity of 73.8% (62.7-83.0) with the same sensitivity and a similar negative predictive value of 96.7%, but a higher positive predictive value of 85.5% (80.3-89.5). Conclusions- The EDCT has excellent sensitivity and negative predictive value and could be a valuable diagnostic tool for the diagnosis of TIA.


The development of responsiveness to substance P and glutamate in the spinal motoneurons of rat fetuses.
The development of responsiveness of motoneurons to substance P (SP) and glutamate was investigated in the isolated spinal cord of the rat fetus at embryonic days 13.5-21.5. The motoneurons at embryonic day 14.5 first responded to SP with a slow depolarization. The responsiveness to SP increased by embryonic days 19.5-21.5. In a low Ca2+ solution, responsiveness was reduced after embryonic day 17.5. Glutamate caused a slow depolarization of motoneurons from embryonic day 13.5. The responsiveness increased until embryonic day 17.5 and decreased thereafter.


Reductive activation of nitroaromatics and enzyme inhibition: misonidazole and xanthine oxidase.
Reductive activation of misonidazole and misondiazole acetate, a simple derivative, in the presence of xanthine oxidase causes inactivation of the enzyme. The inactivation is not accompanied by binding of the misonidazole to the enzyme. The nitroreductase activity of xanthine oxidase is inhibited as measured by the reduction of 3,5-dinitrobenzonitrile (DNBN). Cysteine does not appear to protect against the enzyme inactivation by misonidazole but, by itself, cysteine has a strong stimulating effect on the reduction of DNBN. The possible significance of these reactions to the toxicity of misonidazole are discussed.


Impaired endothelial function in persons with obstructive sleep apnoea: impact of obesity.
Obstructive sleep apnoea (OSA) and obesity are both associated with endothelial dysfunction, which precedes the development of atherosclerosis. As obesity is highly prevalent in OSA, we wanted to test the hypothesis that OSA is associated with endothelial dysfunction independently of obesity. Cross-sectional, population-based study. Norwegian university hospital. Seventy-one subjects (median age 44 years, 35% female) were recruited from a population-based study in Norway. Participants were categorised as obese (body mass index (BMI) ≥30 kg/m(2)), non-obese (BMI<30 kg/m(2)) with OSA (apnoea-hypopnoea index (AHI)≥10), or non-obese without OSA (AHI<5). None. Endothelial function measured by brachial artery ultrasound and expressed as percentage of flow-mediated dilation (FMD%). When non-obese subjects without OSA were used as the reference (FMD% (mean±SD) 10.1±6.3), endothelial function was found to be impaired in subjects with OSA (FMD% 6.4±3.2) (p=0.003). FMD% did not differ between obese (6.0±3.4) and non-obese (6.7±3.1) OSA subjects (p=0.3). By univariate linear regression analysis, AHI, BMI, gender and baseline brachial artery diameter were significantly associated with FMD%. When these variables were entered into a multivariate model, only AHI was significantly associated with FMD%. OSA is associated with endothelial dysfunction independently of obesity and conventional risk factors.


Long-term burden of higher body mass index from childhood on adult cardiometabolic biomarkers: A 30-year cohort study.
Data are limited regarding the association between long-term burden of higher body mass index (BMI) from childhood and cardiometabolic biomarkers. A total of 1553 individuals aged 6-15 years, who were examined 4 or more times for BMI since childhood and followed for 30 years were included in our analysis. Total area under the curve (AUCt) and incremental AUC (AUCi) were calculated as the long-term burden and trends of BMI. Cardiometabolic biomarkers including serum uric acid (SUA), fasting blood-glucose (FBG), and triglyceride to high-density lipoprotein cholesterol ratio (TG/HDL-C) were obtained from venous blood samples. The results showed a positive association of BMI AUCt and AUCi with cardiometabolic biomarkers. After adjusting for demographic variables, the AUCt and AUCi of BMI were significantly associated with a higher level of SUA (β = 3.71; 2.87), FBG (β = 0.09; 0.09), and TG/HDL-C (β = 0.14; 0.11). We performed further studies after dividing subjects into four groups according to AUCt and AUCi of BMI by quartiles. Compared with the lowest quartile group, the highest quartile group had significantly increased risk ratios of hyperuricemia (RR = 2.01; 1.74), type 2 diabetes mellitus (RR = 8.18; 3.96), and high-risk TG/HDL-C (RR = 4.05; 3.26). Our study identifies all subjects' BMI growth curve from childhood and indicates that the long-term burden of higher BMI significantly increases the cardiometabolic risk, and the impact of excessive body weight on cardiometabolic health originates in early life. We emphasize the importance of weight control from childhood for cardiometabolic health.


Plant Systems Biology at the Single-Cell Level.
Our understanding of plant biology is increasingly being built upon studies using 'omics and system biology approaches performed at the level of the entire plant, organ, or tissue. Although these approaches open new avenues to better understand plant biology, they suffer from the cellular complexity of the analyzed sample. Recent methodological advances now allow plant scientists to overcome this limitation and enable biological analyses of single-cells or single-cell-types. Coupled with the development of bioinformatics and functional genomics resources, these studies provide opportunities for high-resolution systems analyses of plant phenomena. In this review, we describe the recent advances, current challenges, and future directions in exploring the biology of single-cells and single-cell-types to enhance our understanding of plant biology as a system.


Small heat shock proteins in Drosophila may confer thermal tolerance.
The four small heat shock proteins in Drosophila melanogaster are genetically linked and simultaneously synthesized, both in response to high temperature and, developmentally, during puparium formation. In tissue culture cells their synthesis is inducible by the molting hormone, ecdysterone. We show here that accompanying their induction and accumulation, the cells and animals acquire thermal tolerance.


Atmospheric oxygen inhibits growth and differentiation of marrow-derived mouse mesenchymal stem cells via a p53-dependent mechanism: implications for long-term culture expansion.
Large scale expansion of human mesenchymal stem cells (MSCs) is routinely performed for clinical therapy. In contrast, developing protocols for large scale expansion of primary mouse MSCs has been more difficult due to unique aspects of rodent biology. Currently, established methods to isolate mouse MSCs select for rapidly dividing subpopulations that emerge from bone marrow cultures following long-term (months) expansion in atmospheric oxygen. Herein, we demonstrate that exposure to atmospheric oxygen rapidly induced p53, TOP2A, and BCL2-associated X protein (BAX) expression and mitochondrial reactive oxygen species (ROS) generation in primary mouse MSCs resulting in oxidative stress, reduced cell viability, and inhibition of cell proliferation. Alternatively, procurement and culture in 5% oxygen supported more prolific expansion of the CD45(-ve) /CD44(+ve) cell fraction in marrow, produced increased MSC yields following immunodepletion, and supported sustained MSC growth resulting in a 2,300-fold increase in cumulative cell yield by fourth passage. MSCs cultured in 5% oxygen also exhibited enhanced trilineage differentiation. The oxygen-induced stress response was dependent upon p53 since siRNA-mediated knockdown of p53 in wild-type cells or exposure of p53(-/-) MSCs to atmospheric oxygen failed to induce ROS generation, reduce viability, or arrest cell growth. These data indicate that long-term culture expansion of mouse MSCs in atmospheric oxygen selects for clones with absent or impaired p53 function, which allows cells to escape oxygen-induced growth inhibition. In contrast, expansion in 5% oxygen generates large numbers of primary mouse MSCs that retain sensitivity to atmospheric oxygen, and therefore a functional p53 protein, even after long-term expansion in vitro.


A snare for the endoscopic extraction of coins.
A special snare is described, by which especially coins can be easily removed from the upper GI tract of children and adults without general anesthesia.


Identification, evolution and expression analyses of mapk gene family in Japanese flounder (Paralichthys olivaceus) provide insight into its divergent functions on biotic and abiotic stresses response.
Mitogen-activated protein kinase (MAPK) are a series of serine/threonine protein kinases showing evolutionary conservation, which can be activated by many stimulus signals and then transfer them from cell membrane to nucleus. MAPKs regulate a variety of biological processes, such as apoptosis, hormone signaling and immune response. In this study, 14 putative mapk genes in Japanese flounder were identified, and their basic physical and chemical properties were characterized. Phylogenetic analysis showed that mapk genes were divided into three main subfamilies, including ERK, JNK and the p38 MAPK. Selection pressure analysis revealed they were evolutionarily-constrained and undergone strong purifying selection. Gene structure and conserved protein motif comparison suggested high levels of conservation in members of mapk gene family. The expression patterns were further investigated in each embryonic and larval development stages and different tissues. In addition, RNA-seq analyses after bacteria and temperature stresses suggested mapk genes had different expression patterns. Three mapk genes showed significant differences in response to E. tarda challenge and five were induced significantly after temperature stress, indicating their potential functions. This systematic analysis provided valuable information for further understanding of the regulation mechanism of mapk gene family under different stresses in Japanese flounder.


Environmental stresses induce the expression of putative glycine-rich insect cuticular protein genes in adult Leptinotarsa decemlineata (Say).
The deposition of cuticular proteins in insects usually occurs during the moulting process. Three putative glycine-rich insect cuticular proteins, Ld-GRP1 to 3, were identified and characterized from the Colorado potato beetle, Leptinotarsa decemlineata. The Ld-GRPs contained conserved GXGX and/or GGXG sequence repeats. Ld-GRP1 also contained a conserved AAPA/V motif commonly found in cuticular proteins. The transcripts of Ld-GRP1 and Ld-GRP2 were detected in the epidermal cell layer by in situ hybridization, making them putative insect cuticular proteins. The putative cuticular protein genes were highly induced by the insecticide azinphosmethyl (organophosphorous) 2-3 weeks after adult moulting. Putative cuticular protein gene expression level was higher in azinphosmethyl-resistant beetles than in susceptible beetles. Furthermore, two of the putative cuticular protein genes were highly induced by dry environmental conditions. These results suggest that the insect might increase cuticular component deposition in the adult stage in response to environmental stresses. This ability may allow the insect to adapt to new or changing environments.


Cholecystokinin-58 and cholecystokinin-8 produce similar but not identical activations of myenteric plexus and dorsal vagal complex.
The enteric nervous system (ENS: myenteric and submucosal plexuses) of the gastrointestinal tract may have a role in the reduction of food intake by cholecystokinin (CCK). Exogenous cholecystokinin-8 (CCK-8) activates the myenteric plexus and the feeding control areas of the dorsal vagal complex (DVC) of the brainstem. An increasing number of reports, however, have shown that CCK-58 is the sole or the major circulating form of CCK in rat, human and dog, and that it is qualitatively different from CCK-8 in evoking various gastrointestinal physiological responses (e.g., contraction of the gallbladder and exocrine pancreatic secretion). In the current report, we compared the abilities of exogenous CCK-58 to activate the myenteric plexus and the dorsal vagal complex with those of exogenous CCK-8 by quantifying Fos-like immunoreactivity (Fos-LI; a marker for neuronal activation). We report that CCK-58 (1, 3, and 5 nmol/kg) increased Fos-LI in the myenteric plexus (p<0.001) and in the DVC (p<0.001) compared to the saline vehicle. The highest dose of CCK-58 increased Fos-LI more than an equimolar dose of CCK-8 in the myenteric plexus and the area postrema. Thus, CCK-8 and CCK-58 produce the same qualitative pattern of activation of central and peripheral neurons, but do not provoke identical quantitative patterns at higher doses. The different patterns produced by the two peptides at higher doses, in areas open to the circulation (myenteric plexus and area postrema) may reflect endocrine actions not observed at lower doses.


Quantifying spatiotemporal heterogeneity of MERS-CoV transmission in the Middle East region: A combined modelling approach.
MERS coronavirus cases notified in the Middle East region since the identification of the virus in 2012 have displayed variations in time and across geography. Through a combined modelling approach, we estimate the rates of generation of cases along the zoonotic and human-to-human transmission routes and assess their spatiotemporal heterogeneity. We consider all cases notified to WHO from March 2012 to mid-September 2014. We use a stochastic modelling of the time series of case incidence in the Middle East region to estimate time- and space-dependent zoonotic and human-to-human transmission parameters. The model also accounts for possible lack of identification of secondary transmissions among notified cases. This approach is combined with the analysis of imported cases out of the region to assess the rate of underreporting of cases. Out of a total of 32 possible models, based on different parameterisation and scenario considered, the best-fit model is characterised by a large heterogeneity in time and across space for both zoonotic and human-to-human transmission. The variation in time that occurred during Spring 2014 led to a 17-fold and 3-fold increase in the two transmissions, respectively, bringing the reproductive rate to values above 1 during that period for all regions under study. The model suggests that 75% of MERS-CoV cases are secondary cases (human-to-human transmission), which is substantially higher than the 34% of reported cases with an epidemiological link to another case. Overall, estimated reporting rate is 0.26. Our findings show a higher level of spatial heterogeneity in zoonotic transmission compared to human-to-human, highlighting the strong environmental component of the epidemic. Since sporadic introductions are predicted to be a small proportion of notified cases and are responsible for triggering secondary transmissions, a more comprehensive understanding of zoonotic source and path of transmission could be critical to limit the epidemic spread.


Traditional Chinese medicine for the prevention and treatment of COVID-19: A protocol for systematic review and network meta-analysis.
The coronavirus disease 2019 (COVID-19) pandemic spread to most of the world's countries during its first and subsequent waves, often increasing in an almost exponential trend. Traditional Chinese medicine has played a vital role in this epidemic. Nonetheless, size of effect, certainty of the evidence, optimal therapy regimen, and selection of patients who are likely to benefit most are factors that remain to be evaluated. This study aims to assess and rank where appropriate the relative effects of interventions for the prevention and treatment of COVID-19. This study will follow the Preferred Reporting Items for Systematic Reviews and Meta-analysis Protocols. We will search Chinese electronic database (CBM, Wanfang and CNKI) and international electronic databases (PubMed, Embase, Cochrane Library, and Web of Science) for identify all relevant published studies. Study selection, data collection and assessment of study bias will be conducted independently by a pair of independent reviewers. The Cochrane risk of bias tool will be used for the risk of bias assessment. We will use the advance of GRADE to rate the certainty of network meta-analysis. Data analysis will be performed with R-3.6.1 and WinBUGS software. The results of this study will be published in a peer-reviewed journal. This systematic review and network meta-analysis will use both direct and indirect evidence to compare the differences of all Traditional Chinese medicine treatment for COVID-19 patients, providing decision-makers and clinical practitioners with a complete, high-quality and up-to-date synthesis of evidence.


Protein-Ligand Binding Molecular Details Revealed by Terahertz Optical Kerr Spectroscopy: A Simulation Study.
Picosecond fast motions and their involvement in the biochemical processes such as protein-ligand binding has engaged significant attention. Terahertz optical Kerr spectroscopy (OKE) has the superior potential to probe these fast motions directly. Application of OKE in protein-ligand binding study is, however, limited by the difficulty of quantitative atomistic interpretation, and the calculation of Kerr spectrum for entire solvated protein complex was considered not yet feasible, due to the lack of one consistent polarizable model for both configuration sampling and polarizability calculation. Here, we analyzed the biochemical relevance of OKE to the lysozyme-triacetylchitotriose binding based on the first OKE simulation using one consistent Drude polarizable model. An analytical multipole and induced dipole scheme was employed to calculate the off-diagonal Drude polarizability more efficiently and accurately. Further theoretical analysis revealed how the subtle twisting and stiffening of aromatic protein residues' spatial arrangement as well as the confinement of small water clusters between ligand and protein cavity due to the ligand binding can be examined using Kerr spectroscopy. Comparison between the signals of bound complex and that of uncorrelated protein/ligand demonstrated that binding action alone has reflection in the OKE spectrum. Our study indicated OKE as a powerful terahertz probe for protein-ligand binding chemistry and dynamics.


Emotional state and cancer-related self-efficacy as affecting resilience and quality of life in kidney cancer patients: a cross-sectional study.
To assess the relationship between resilience and quality of life (QOL) of kidney cancer patients, including influencing factors. Based on a cross-sectional study design, participants (N = 103) were recruited from patients who were admitted to the urology clinic of a medical center in Taiwan between April 2020 and January 2021. Data collection was accomplished via a questionnaire. The study variables included demographic information, disease attributes, happiness level, depression, cancer-related self-efficacy, resilience, and QOL. One-way analysis of variance, Pearson correlation coefficients, independent-sample t-tests, hierarchical regression, and process analysis were the statistical methods used to analyze the data. Kidney cancer patients who were less depressed exhibited better cancer-related self-efficacy and have better resilience. In non-depressed individuals, higher levels of happiness and better resilience resulted in better QOL. Resilience is a mediator that affects the relationship between depression and QOL. Patients with better emotional state experience better resilience and QOL. Patients' better cancer-related self-efficacy is related to better resilience while better resilience is associated with better QOL. Clinical care providers need to evaluate and improve cancer-related self-efficacy, emotional state, and resilience of kidney cancer patients, which will improve their resilience and QOL.


Links between mothers' coping styles, toddler reactivity, and sensitivity to toddler's negative emotions.
The extent to which engaged maternal coping styles moderate the association between toddler's temperamental reactivity and mothers' sensitivity to children's negative emotions was examined in 89 mother-child dyads. Primiparous mothers completed a measure of coping styles prenatally. When toddlers were 16 months old, mothers completed a measure of perceived toddler temperament and a self-report of how they respond to toddler negative emotions, and maternal sensitivity and temperamental reactivity were observed during emotionally arousing tasks in the laboratory. Mothers' disengaged coping style was positively associated with self-reported insensitive responses to children's negative emotions. Engaged coping moderated the association between toddler temperamental reactivity and both self-reported insensitive responses and observed maternal sensitivity, such that temperamental reactivity was more strongly linked with less sensitive maternal behavior when engaged coping was low.


Optimization of xylanase production by filamentous fungi in solid-state fermentation and scale-up to horizontal tube bioreactor.
Five microorganisms, namely Aspergillus niger CECT 2700, A. niger CECT 2915, A. niger CECT 2088, Aspergillus terreus CECT 2808, and Rhizopus stolonifer CECT 2344, were grown on corncob to produce cell wall polysaccharide-degrading enzymes, mainly xylanases, by solid-state fermentation (SSF). A. niger CECT 2700 produced the highest amount of xylanases of 504 ± 7 U/g dry corncob (dcc) after 3 days of fermentation. The optimization of the culture broth (5.0 g/L NaNO3, 1.3 g/L (NH4)2SO4, 4.5 g/L KH2PO4, and 3 g/L yeast extract) and operational conditions (5 g of bed loading, using an initial substrate to moistening medium of 1:3.6 (w/v)) allowed increasing the predicted maximal xylanase activity up to 2,452.7 U/g dcc. However, different pretreatments of materials, including destarching, autoclaving, microwave, and alkaline treatments, were detrimental. Finally, the process was successfully established in a laboratory-scale horizontal tube bioreactor, achieving the highest xylanase activity (2,926 U/g dcc) at a flow rate of 0.2 L/min. The result showed an overall 5.8-fold increase in xylanase activity after optimization of culture media, operational conditions, and scale-up.


Effectiveness of washed platelet concentrate and red cell transfusions for a patient with anhaptoglobinemia with antihaptoglobin antibody.
A 71-year-old Japanese male with myelodysplastic syndrome progressing to overt leukaemia and hepatocellular carcinoma developed dyspnea and urticaria immediately after infusion of platelet concentrate (PC). He exhibited an identical reaction following blood transfusion. Serum haptoglobin was undetectable. The patient was determined to be homozygous for Hp(del) by polymerase chain reaction (PCR). Antibody to haptoglobin was detected by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. No antibodies against human leucocyte antigen (HLA) or platelet-specific antigens were detected. Washed PC and washed red blood cells were effective in preventing the transfusion-related anaphylactoid reactions.


Longitudinal changes in growth hormone response to growth hormone-releasing hormone in neonatal rhesus monkeys.
To determine whether differential response to growth hormone-releasing hormone (GHRH) could cause the developmental changes seen in growth hormone (GH) secretion, we administered 10 micrograms/kg GHRH (1-44 NH2) to a group of four unanesthetized, fasted, rhesus monkeys via acutely placed venous catheters at 1, 7, 14, and 28 d postnatal age. Serum GH was assayed by hGH RIA in sera collected at -60, -30, 0, 15, 30, 45, 60, and 90 min relative to the GHRH bolus. Serum cortisol was measured by ELISA in the 0-, 30-, and 60-min samples. Differences between age groups were analyzed by repeated measures analysis of variance and paired t tests. Mean basal GH levels were higher at 1 d (9.4 +/- 1.2 micrograms/L, mean +/- SEM) than at 7 (5.5 +/- 0.4), 14 (5.6 +/- 0.5), and 28 d (5.3 +/- 0.5) of age. There were no other significant differences in mean basal GH values between the age groups. Mean post-GHRH GH concentrations decreased significantly with each age after 1 d (22.6 +/- 1.6): 7 d (16.4 +/- 1.3); 14 d (11.3 +/- 1.0); and 28 d (7.9 +/- 0.9). Similarly, mean delta-GH values decreased with each increase in age from 1 d (15.0 +/- 1.9): 7 d (10.9 +/- 1.6); 14 d (5.9 +/- 1.1); and 28 d (2.7 +/- 0.8). Serum cortisol was not correlated with serum GH at any age. Our study demonstrates decreasing basal GH concentration and GH responses to GHRH with advancing age from 1 to 28 d in the rhesus monkey.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular cloning of cDNA for rat acyl-CoA oxidase.
Poly(A+) RNA was prepared from hepatic free polysomes of rats which had been fed di(2-ethylhexyl) phthalate for the induction of peroxisomal beta-oxidation enzymes. This preparation was enriched for the mRNAs of these enzymes by sucrose density gradient centrifugation, and used for the synthesis of double-stranded cDNA. Recombinant plasmids were constructed from the cDNA and pBR322 by dG X dC-tailing method and used for the transformation of an Escherichia coli strain, chi 1776. By differential colony hybridization using [32P]cDNA of partially purified liver poly(A+) RNA from induced and noninduced rats as probes, and then by hybridization-selected translation, we obtained two clones with cDNA inserts which specifically selected acyl-CoA oxidase mRNA. On Northern blotting, both cDNA inserts hybridized to 3.8-kilobase RNA which was increased about 10-fold by di(2-ethylhexyl) phthalate treatment of the rats. The cleavage maps of the cDNA inserts showed they overlap with each other. We conclude that the above two recombinant plasmid clones contain cDNA sequences for rat acyl-CoA oxidase.


Plasma creatine kinase isozyme electrophoretic pattern of long-term vitamin E-deficient rats.
Weanling male Sprague Dawley rats were fed a vitamin E-deficient basal diet with or without supplementation of 100 IU/kg diet of vitamin E for 12 months. The isozyme pattern of creatine kinase (CPK) in plasma was determined electrophoretically. The CPK activity of the plasma of control rats averaged 43.2 nmoles/min/ml and was predominantly CPK3 isozyme, while that of the plasma of vitamin E-deficient rats averaged 100.8 nmoles/min/ml and consisted of CPK2 and CPK3 isozymes. The results suggest that the increased CPK activity in the plasma of vitamin E-deficient rats may originate from heart and skeletal muscle degeneration.


Amphetamine increases activity but not exploration in humans and mice.
Cross-species quantification of physiological behavior enables a better understanding of the biological systems underlying neuropsychiatric diseases such as bipolar disorder (BD). Cardinal symptoms of manic BD include increased motor activity and goal-directed behavior, thought to be related to increased catecholamine activity, potentially selective to dopamine homeostatic dysregulation. The objective of this study was to test whether acute administration of amphetamine, a norepinephrine/dopamine transporter inhibitor and dopamine releaser, would replicate the profile of activity and exploration observed in both humans with manic BD and mouse models of mania. Healthy volunteers with no psychiatric history were randomized to a one-time dose of placebo (n = 25), 10 mg d-amphetamine (n = 18), or 20 mg amphetamine (n = 23). Eighty mice were administered one of four doses of d-amphetamine or vehicle. Humans and mice were tested in the behavioral pattern monitor (BPM), which quantifies motor activity, exploratory behavior, and spatial patterns of behavior. In humans, the 20-mg dose of amphetamine increased motor activity as measured by acceleration without marked effects on exploration or spatial patterns of activity. In mice, amphetamine increased activity, decreased specific exploration, and caused straighter, one-dimensional movements in a dose-dependent manner. Consistent with mice, amphetamine increased motoric activity in humans without increasing exploration. Given that BD patients exhibit heightened exploration, these data further emphasize the limitation of amphetamine-induced hyperactivity as a suitable model for BD. Further, these studies highlight the utility of cross-species physiological paradigms in validating biological mechanisms of psychiatric diseases.


Ketamine for acute neuropathic pain in patients with spinal cord injury.
Ketamine, an N-methyl-d-aspartic acid (NMDA) receptor antagonist, may be useful for treating neuropathic pain, which is often difficult to control. We report a prospective study of 13 patients with acute neuropathic pain due to spinal cord injury (SCI) treated with ketamine. All underwent a test challenge with 5mg ketamine. Patients with satisfactory responses were then treated intravenously and subsequently perorally with ketamine. Pre- and post-treatment pain was recorded on a visual analogue scale. All 13 patients responded positively to the ketamine test challenge and underwent continued ketamine administration. At the cessation of treatment and alter at final follow up, pain was decreased by 74.7% and 96.8%, respectively. The average administration period was 17.2 days; it was longer (59 days) in one patient treated in the subacute phase. All patients suffered allodynia-type pain and experienced 30% or less of their original pain intensity upon test challenge. Side effects were noted in five patients, although their severity did not require treatment cessation. In patients with SCI, ketamine reduced allodynia. Particularly good results were obtained in patients treated in the acute phase and these patients did not experience post-treatment symptom recurrence. Our results suggest that in patients with SCI, ketamine is useful for treating neuropathic pain in the acute phase.


Transforming genetic research practices with marginalized communities: a case for responsive justice.
Genetics researchers often work with distinct communities. To take moral account of how their research affects these communities, they need a richer conception of justice and they need to make those communities equal participants in decision-making about how the research is conducted and what is produced and published out of it.


Simultaneous quantitation of urinary cotinine and acrylonitrile-derived mercapturic acids with ultraperformance liquid chromatography-tandem mass spectrometry.
Acrylonitrile (AN), a widely used industrial chemical also found in tobacco smoke, has been classified as a possible human carcinogen (group 2B) by the International Agency for Research on Cancer. AN can be detoxified by glutathione S-transferase (GST) to form glutathione (GSH) conjugates in vivo. It can be metabolically activated by cytochrome P450 2E1 to form 2-cyanoethylene oxide, which can also be detoxified by GST to generate GSH conjugates. The GSH conjugates can be further metabolized to mercapturic acids (MAs), namely, N-acetyl-S-(2-cyanoethyl)cysteine (CEMA), N-acetyl-S-(2-hydroxyethyl)cysteine (HEMA), and N-acetyl-S-(1-cyano-2-hydroxyethyl)cysteine (CHEMA). This study developed an ultraperformance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method to quantitatively profile the major AN urinary metabolites (CEMA, HEMA, and CHEMA) to assess AN exposure, as well as analyze urinary cotinine (COT) as an indicator for tobacco smoke exposure. The limits of quantitation were 0.1, 0.1, 1.0, and 0.05 μg/L for HEMA, CEMA, CHEMA, and COT, respectively. This method was applied to analyze the three AN-derived MAs in 36 volunteers with no prior occupational AN exposure. Data analysis showed significant correlations between the level of COT and the levels of these MAs, suggesting them as biomarkers for exposure to low levels of AN. The results demonstrate that a highly specific and sensitive UPLC-MS/MS method has been successfully developed to quantitatively profile the major urinary metabolites of AN in humans to assess low AN exposure.


Vemurafenib activates the sonic hedgehog pathway and promotes thyroid cancer stem cell self-renewal.
B-Raf kinase inhibitors such as vemurafenib (PLX4032) and dabrafenib have limited therapeutic efficacy on BRAF-mutated thyroid cancer. Cancer stem cells (CSCs) play important roles in tumor recurrence, drug resistance, and metastasis. Whether CSCs play a role in dampening the antitumor activity of B-Raf kinase inhibitors remains unknown. Here, we report that vemurafenib (PLX4032) induced the expression of several stemness-related genes including Gli1, Snail, BMI1, and SOX2 in two anaplastic thyroid cancer cell lines, SW1736 and 8505C, but decreased the expression of these genes in A375 cells, a human melanoma cell line. PLX4032 promoted thyroid cancer stem cell self-renewal, as evidenced by increased numbers of aldehyde dehydrogenase-positive cells and thyrospheres. Mechanistically, PLX4032 activates the PI-3 and mitogen-activated protein kinase pathways through HER3 to cross-activate Gli1, a transcription factor of the sonic hedgehog (Shh) pathway. GANT61, a specific inhibitor of Gli1, blocked the expression of the stemness-related genes in PLX4032-treated thyroid cancer cells in vitro and in vivo in two thyroid cancer xenograft models. GANT61 treatment alone weakly inhibited SW1736 tumor growth but enhanced the antitumor activity of PLX4032 when used in combination. Our study provides mechanistic insights into how thyroid cancer poorly responds to B-Raf kinase inhibitors and suggests that targeting B-Raf and the Shh pathway in combination may overcome thyroid cancer drug resistance.


Cardiovascular effects of ethanol in anaesthetized, conscious and pithed rats.
The influence of ethanol (2.0 g/kg p.o.) on heart rate and blood pressure in anaesthetized, conscious and pithed rats was studied. In anaesthetized rats we observed an increase in heart rate and a marked and very sudden decrease in blood pressure. In pithed rats blood pressure decreased more slowly than in anaesthetized and conscious animals and there were no changes in heart rate. These results show that the central nervous system may be responsible for the increase in heart rate and early phase of hypotension caused by acute ethanol administration. The slower decrease (up to 1 h) in blood pressure may be caused by central and different indirect peripheral mechanisms.


Role of mitochondrial uncoupling protein-2 (UCP2) in higher brain functions, neuronal plasticity and network oscillation.
Major psychiatric illnesses, affecting 36% of the world's population, are profound disorders of thought, mood and behavior associated with underlying impairments in synaptic plasticity and cellular resilience. Mitochondria support energy demanding processes like neural transmission and synaptogenesis and are thus points of broadening interest in the energetics underlying the neurobiology of mental illness. These experiments interrogated the importance of mitochondrial flexibility in behavior, synaptic and cortical activity in a mouse model. We studied mice with ablated uncoupling protein-2 expression (UCP2 KO) and analyzed cellular, circuit and behavioral attributes of higher brain regions. We found that mitochondrial impairment induced by UCP2 ablation produces an anxiety prone, cognitively impaired behavioral phenotype. Further, NMDA receptor blockade in the UCP2 KO mouse model resulted in changes in synaptic plasticity, brain oscillatory and sensory gating activities. We conclude that disruptions in mitochondrial function may play a critical role in pathophysiology of mental illness. Specifically, we have shown that NMDA driven behavioral, synaptic, and brain oscillatory functions are impaired in UCP2 knockout mice.


Carnoy solution versus GEWF solution for lymph node revealing in colorectal cancer: a randomized controlled trial.
This study aimed to compare the performance of two lymph node revealing solutions. This randomized clinical trial (NTC02704988) investigated patients with colon or rectal cancer who underwent surgical resection with D2 lymphadenectomy. Specimens submitted for conventional pathological examination were randomly assigned for additional fixation with Carnoy or GEWF solution, and dissection was performed to examine the missed lymph nodes. The number of lymph nodes retrieved, additional identified metastatic lymph nodes, lymph node upstaging, and complementary indication of adjuvant therapy were investigated. The number of lymph nodes retrieved was significantly higher with the use of lymph node revealing solutions than with the conventional method in colon cancer (GEWF: 29.5 vs 27; p < 0.001; Carnoy: 27.7 vs 25.2; p < 0.001) and rectal cancer (GEWF: 25.8 vs 23.6; p < 0.001; Carnoy: 23.1 vs 20.8; p < 0.001). There were no differences between the solutions and conventional examination with respect to the median number of additional metastatic lymph nodes identified (0 in all arms), the number of patients with lymph node upstaging (colon cancer: 1 in the Carnoy arm, 0 in the GEWF arm; rectal cancer: 1 in the GEWF arm, 0 in the Carnoy arm), or the number of patients with complementary indication of adjuvant therapy (colon cancer: 1 in the Carnoy arm, 0 in the GEWF arm; rectal cancer: 0 in both arms). Despite the higher number of lymph nodes retrieved, neither solution resulted in significant changes in patient staging or treatment. Both solutions exhibited equal performance with respect to all outcomes. NTC02704988.


Sex-dependent effects of periadolescent exposure to the cannabinoid agonist CP-55,940 on morphine self-administration behaviour and the endogenous opioid system.
Early cannabinoid consumption may predispose individuals to the misuse of addictive drugs later in life. However, there is a lack of experimental evidence as to whether cannabinoid exposure during adolescence might differently affect opiate reinforcing efficacy and the opioid system in adults of both sexes. Our aim was to examine whether periadolescent chronic exposure to the cannabinoid agonist CP-55,940 could exert sex-dependent effects on morphine reinforcing and the opioid system in adulthood. Morphine reinforcing was studied under a progressive ratio (PR) reinforcement schedule in adult male and female rats that previously acquired morphine self-administration under a fixed ratio 1 (FR1) schedule. Binding levels and functionality of mu-opioid receptors were also evaluated. Periadolescent cannabinoid exposure altered morphine self-administration and the opioid system in adult rats in a sex-dependent manner. CP-55,940-exposed males exhibited higher self-administration rates under a FR1, but not under a PR schedule. In females, CP-55,940 did not modify morphine self-administration under either schedule. Moreover, CP-55,940 also increased mu-opioid receptor levels in the subcallosal streak of pre-treated animals and decreased mu-opioid receptor functionality in the nucleus accumbens shell but again, only in males. Our data indicate that adult male rats exposed to the cannabinoid in adolescence self-administer more morphine than females, but only when the demands required by the schedule of reinforcement are low, which might be related to the decrease in mu-opioid receptor functionality in the NAcc-shell observed in these animals.


Development in vitro of cytotoxic lymphocytes against murine cytomegalovirus.
Lymphocytes cytotoxic for mouse embryo fibroblasts (MEF) infected with murine cytomegalovirus (MCMV) were produced by in vitro culture of "memory" spleen cells with UV-irradiated, MCMV-infected, MEF. Cytotoxic lymphocytes were developed from spleen cells of mice 10 to 240 days after infection with MCMV. The cytotoxic cells carried the theta and Ly 2 antigens, and were H-2 restricted in the recognition of infected target cells.


Dilatation and curettage fails to detect most focal lesions in the uterine cavity in women with postmenopausal bleeding.
To determine the prevalence of focally growing lesions in the uterine cavity in women with postmenopausal bleeding and endometrium > or = 5 mm and the extent to which such lesions can be correctly diagnosed by D&C. In a prospective study, 105 women with postmenopausal bleeding and endometrium > or = 5 mm at transvaginal ultrasound examination underwent diagnostic hysteroscopy, D&C and hysteroscopic resection of any focally growing lesion still left in the uterine cavity after D&C. Twenty-four women also underwent hysterectomy. If the histological diagnosis differed between specimens from the same patient, the most relevant diagnosis was considered the final one. Eighty percent (84/105) of the women had pathology in the uterine cavity, and 98% (82/84) of the pathological lesions manifested a focal growth pattern at hysteroscopy. In 87% of the women with focal lesions in the uterine cavity, the whole or parts of the lesion remained in situ after D&C. D&C missed 58% (25/43) of polyps, 50% (5/10) of hyperplasias, 60% (3/5) of complex atypical hyperplasias, and 11% (2/19) of endometrial cancers. The agreement between the D&C diagnosis and the final diagnosis was excellent (94%) in women without focally growing lesions at hysteroscopy. If there are focal lesions in the uterine cavity, hysteroscopy with endometrial resection is superior to D&C for obtaining a representative endometrial sample in women with postmenopausal bleeding and endometrium > or = 5 mm.


Fulminant severe retinopathy in a newly diagnosed diabetic without risk factors.
The authors describe a 26-year-old female developing severe proliferative retinopathy within 2 weeks of diagnosis of insulin-dependent diabetes mellitus. The patient presented with profound diabetic ketoacidosis, but had no other risk factors for retinopathy. She had marked bilateral myopia which did not protect her from retinopathy. Nine months after diagnosis of diabetes, she has profound visual impairment despite correction for myopia, and has also developed marked autonomic neuropathy and diabetic nephropathy.


Analysis of simplicial complexes to determine when to sample for quantitative DCE MRI of the breast.
A method is presented to select the optimal time points at which to measure DCE-MRI signal intensities, leaving time in the MR exam for high-spatial resolution image acquisition. Simplicial complexes are generated from the Kety-Tofts model pharmacokinetic parameters Ktrans and ve . A geometric search selects optimal time points for accurate estimation of perfusion parameters. The DCE-MRI data acquired in women with invasive breast cancer (N = 27) were used to retrospectively compare parameter maps fit to full and subsampled time courses. Simplicial complexes were generated for a fixed range of Kety-Tofts model parameters and for the parameter ranges weighted by estimates from the fully sampled data. The largest-area manifolds determined the optimal three time points for each case. Simulations were performed along with retrospectively subsampled data fits. The agreement was computed between the model parameters fit to three points and those fit to all points. The optimal three-point sample times were from the data-informed simplicial complex analysis and determined to be 65, 204, and 393 s after arrival of the contrast agent to breast tissue. In the patient data, tumor-median parameter values fit using all points and the three selected time points agreed with concordance correlation coefficients of 0.97 for Ktrans and 0.67 for ve . It is possible to accurately estimate pharmacokinetic parameters from three properly selected time points inserted into a clinical DCE-MRI breast exam. This technique can provide guidance on when to capture images for quantitative data between high-spatial-resolution DCE-MRI images.


Multiple functions of Hes genes in the proliferation and differentiation of neural stem cells.
The HES proteins (hairy and Enhancer of split (E(spl)) homologs) are basic helix-loop-helix (bHLH) transcription factors that regulate the proliferation and differentiation of stem cells. Family members HES1, 3, and 5 are all critical regulators of nervous system development. The Hes genes exhibit oscillatory expression levels, and this dynamic expression allows for the complex regulation of numerous downstream genes such as Ascl1, Neurog2, Olig2 involved in the differentiation of specific cell types. In addition, HES proteins act as hubs for the molecule crosstalk among Notch, Wnt, and other signaling pathways that regulate nervous system development.


A Random Matrix Theory Approach to Denoise Single-Cell Data.
Single-cell technologies provide the opportunity to identify new cellular states. However, a major obstacle to the identification of biological signals is noise in single-cell data. In addition, single-cell data are very sparse. We propose a new method based on random matrix theory to analyze and denoise single-cell sequencing data. The method uses the universal distributions predicted by random matrix theory for the eigenvalues and eigenvectors of random covariance/Wishart matrices to distinguish noise from signal. In addition, we explain how sparsity can cause spurious eigenvector localization, falsely identifying meaningful directions in the data. We show that roughly 95% of the information in single-cell data is compatible with the predictions of random matrix theory, about 3% is spurious signal induced by sparsity, and only the last 2% reflects true biological signal. We demonstrate the effectiveness of our approach by comparing with alternative techniques in a variety of examples with marked cell populations.


Esophageal manometry in 95 healthy adult volunteers. Variability of pressures with age and frequency of "abnormal" contractions.
Although esophageal manometry is widely used in clinical practice, the normal range of esophageal contraction parameters is poorly defined. Therefore, 95 healthy volunteers (mean age: 43 years; range 22-79 years) were studied with a low-compliance infusion system and 4.5-mm-diameter catheter. All subjects were given 10 wet swallows (5 cc H2O) and 38 subjects also were given 10 dry swallows. Amplitude, but not duration, was greater (P less than 0.05) after wet compared to dry swallows. Both distal mean contractile amplitude and duration of wet swallows significantly increased with age and peaked in the fifties. Double-peaked waves frequently occurred after both wet (11.3%) and dry (18.1%) swallows, but triple-peaked waves were rare (less than 1%). Nonperistaltic contractions were more common (P less than 0.001) after dry compared to wet swallows (18.1% vs 4.1%). This difference resulted from frequent simultaneous contractions after dry swallows (12.6% vs 0.4%). Distal esophageal contractile amplitude and duration after wet swallows increases with age. Triple-peaked waves and wet-swallow-induced simultaneous contractions should suggest an esophageal motility disorder. Double-peaked waves are a common variant of normal. Dry swallows have little use in the current evaluation of esophageal peristalsis.


Analysis of arsenic and calcium in soil samples by laser ablation mass spectrometry.
We present an analytical procedure based on laser ablation mass spectrometry (LAMS) in order to detect and quantify arsenic and calcium in soil samples and we analyze the diverse factors that influence the precision of LAMS, such as laser fluence and matrix effect. The results indicate that a Zn matrix is a good choice for the analysis of those metals in soil samples. This work also provides a method for the direct determination of As in soil samples whose concentrations are lower than 100 ppm with a 70 ppm minimum detection limits (MDL).


Comparative ecotoxicity of graphene, functionalized multi-walled CNTs, and their mixture in freshwater microalgae, Scenedesmus obliquus: analyzing the role of oxidative stress.
Due to their remarkable properties, the applications of carbon-based nanomaterials (CNMs) such as graphene and functionalized multi-walled carbon nanotubes (f-MWCNTs) are increasing. These CNMs can enter the freshwater environment via numerous routes, potentially exposing various organisms. The current study assesses the effects of graphene, f-MWCNTs, and their binary mixture on the freshwater algal species Scenedesmus obliquus. The concentration for the individual materials was kept at 1 mg L-1, while graphene and f-MWCNTs were taken at 0.5 mg L-1 each for the combination. Both the CNMs caused a decrease in cell viability, esterase activity, and photosynthetic efficiency in the cells. The cytotoxic effects were accompanied by increased hydroxyl and superoxide radical generation, lipid peroxidation, antioxidant enzyme activity (catalase and superoxide dismutase), and mitochondrial membrane potential. Graphene was more toxic compared to f-MWCNTs. The binary mixture of the pollutants demonstrated a synergistic enhancement of the toxic potential. Oxidative stress generation played a critical role in toxicity responses, as noted by a strong correlation between the physiological parameters and the biomarkers of oxidative stress. The outcomes from this study emphasize the significance of considering the combined effects of various CNMs as part of a thorough evaluation of ecotoxicity in freshwater organisms.


Long-term management of adults with conotruncal lesions: the diagnostic approach at All Children's Hospital.
Improved survival in children with complex congenital cardiac disease, such as conotruncal abnormalities, has created a sub-population of children and young adults who need comprehensive multi-disciplinary long-term follow-up. Routine surveillance with comprehensive screening for structural heart disease, functional heart disease, thromboembolic disease, arrhythmias, and associated end-organ dysfunction is important. Future research will better define the care plans for routine surveillance in patients with conotruncal abnormalities.


The effect of protein and calorie intake on prealbumin, complications, length of stay, and function in the acute rehabilitation inpatient with stroke.
Nutrition's impact on stroke rehabilitation outcomes is controversial. Existing studies utilize albumin without correcting for inflammation in nutritional assessments. Here, prealbumin was used and inflammation assessed to determine if nutrition impacts rehabilitation outcomes. Determine the effect of dietary intake on prealbumin level, number of complications, length of stay, and Functional Independence Measure (FIM) efficiency in rehabilitation stroke inpatients. Patients had admission and discharge prealbumin and C-reactive protein (CRP) levels drawn; and, weekly protein and calorie counts obtained. Patients were followed for number of complications, length of stay, and FIM efficiency. Mean protein and calorie intake was 57.6 ± 16.2 g/d and 1452.2 ± 435.8 kcal/d, respectively. 77.6% of patients had normal prealbumin on admission with 94.9% on discharge. Prealbumin increased significantly from admission to discharge (22.3 ± 6.2 mg/dL vs. 24.6 mg/dL ± 5.1 mg/dL, P = 0.007). Number of complications and length of stay were predicted by CRP in regression models. Total, motor, and cognitive FIM efficiencies were not universally affected by prealbumin levels, protein intake, or calorie intake. Nearly all hypoprealbuminemic stroke rehabilitation inpatients correct their levels eating a non-supplemented diet. Number of complications, length of stay, and functional outcomes in this patient are not affected by prealbumin levels, protein intake, or calorie intake.


SARS-CoV-2 infection induces robust germinal center CD4 T follicular helper cell responses in rhesus macaques.
CD4 T follicular helper (T fh ) cells are important for the generation of durable and specific humoral protection against viral infections. The degree to which SARS-CoV-2 infection generates T fh cells and stimulates the germinal center response is an important question as we investigate vaccine options for the current pandemic. Here we report that SARS-CoV-2 infection resulted in transient accumulation of pro-inflammatory monocytes and proliferating T fh cells with a T h 1 profile in peripheral blood. CD4 helper cell responses were skewed predominantly toward a T h 1 response in blood, lung, and lymph nodes. We observed the generation of germinal center T fh cells specific for the SARS-CoV-2 spike (S) and nucleocapsid (N) proteins, and a corresponding early appearance of antiviral serum IgG antibodies. Our data suggest that a vaccine promoting T h 1-type T fh responses that target the S protein may lead to protective immunity.


Male European eels are highly efficient long distance swimmers: effects of endurance swimming on maturation.
European eels (Anguilla anguilla) migrate ~6000km towards their spawning area in the Sargasso Sea. Based on the recent discovery that males swim even more efficiently than females, it was predicted that males also would be able to swim ~6000km within six months. Additionally, eels do not mature naturally in captivity due to strong neural inhibition. Earlier, it was hypothesized that swimming exercise is a natural trigger to induce sexual maturation and may even result in full maturation. In the present study two groups of farmed male silver eels were subjected to either endurance swimming or resting for up to 6months. It was found that male eels were able to swim continuously for a total distance of 6670km within 6months. The body weight decrease in swimming and resting males after 6months was similar (<30g) underlining the extreme low energy cost of swimming. In contrast to our expectation long-term swimming did not induce sexual maturation in farmed silver eels, suggesting that swimming alone is not sufficient as a trigger for sexual maturation. In conclusion, male eels are efficient long distance swimmers and likely able to cover the distance to the Sargasso Sea within the expected time span of 6months.


Tracheal Injury during Endoscopic Transoral Vestibular Approach Thyroidectomy: A Case Report.
This report describes the intraoperative course of endoscopic thyroidectomy by oral vestibular approach in a female patient. This operation is new, and its perioperative management is not yet mature. In this case, the surgery resulted in trachea injury that could not be detected easily. As a result, the patient suddenly developed acute dyspnea and circulatory disorder. This procedure requires caution in surgical execution and anesthesia management.


Endoprosthetic reconstruction for lower extremity soft tissue sarcomas with bone involvement.
Bone resection and endoprosthetic reconstruction (EPR) in the setting of soft tissue sarcoma (STS) management is rare and incurs unique challenges. We aim to report on the surgical and oncological outcomes of this relatively previously undocumented cohort. This is a single-center retrospective review of prospectively collected data for patients who required EPRs following resection of STSs of the lower extremity. Following inclusion criteria, we assessed 29 cases of EPR for primary STS of the lower limb. The mean age was 54 years (range 18-84). Of the 29 patients, there were 6 total femur, 11 proximal femur, 4 intercalary, and 8 distal femur EPRs. Fourteen of 29 patients (48%) underwent re-operations for surgical complications, with 9 relating to infection (31%). When a matched cohort analysis was performed comparing our cohort to STSs that did not necessitate EPR, a reduced rate of overall survival and metastasis-free survival was found in those requiring EPR. This series identifies a high rate of complication from EPRs performed for STS. Patients should be cautioned about the high rate of infection, surgical complications, and lower overall survival in this setting.


Surgical Exposure of Impacted Teeth.
Impacted incisors, canines, premolars, and second molar are problems encountered frequently by general dentists, orthodontists, and oral and maxillofacial surgeons. The etiology of impacted teeth is multifactorial. Traditional radiographs can be used for location of the impacted tooth but 3-D CBCT is superior in evaluating the tooth's position. Successful management requires an interdisciplinary approach with an orthodontist responsible for the overall success of the treatment plan. Surgical exposure of these impacted teeth is accomplished using an open or closed surgical procedure. Choosing the appropriate surgical procedure and orthodontic treatment plan will result in a stable, predictable, and aesthetic result.


Temporising external fixation of calcaneus fractures prior to definitive plate fixation: a case series.
Surgical management of calcaneus fractures is technically demanding and has a high risk of wound complications. These fractures are traditionally managed with splinting until swelling has subsided, which can take weeks and leaves the fracture fragments displaced. We describe a novel protocol for the management of displaced intraarticular calcaneus fractures that utilises a temporising external fixator and staged conversion to plate fixation through a sinus tarsi approach. The goal of this technique was to enable earlier treatment with open reduction and internal fixation, minimise the amount of manipulation required at the time of definitive fixation and reduce the wound complication rate seen with the traditional extensile approach. The records of patients with displaced calcaneus fractures from 2010-2014 were reviewed retrospectively. A total of nine patients with 10 calcaneus fractures were treated using this protocol. All patients underwent ankle-spanning medial external fixation within 48 hours after injury. Patients underwent conversion to open plate fixation through a sinus tarsi approach when skin turgor had returned to normal. Time to surgery, infection rate, wound complications, radiographic alignment, and time to radiographic union were recorded. The average Bohler's angle improved from 13.2 (range -2 to 34) degrees preoperatively to 34.3 (range 26 to 42) degrees postoperatively. The average time from external fixation to conversion to internal fixation was 4.8 (range 3 to 7) days. There were no immediate post-surgical complications. The average time to weight-bearing was 8.5 weeks. The average time to radiographic union was 9.5 (range 8 to 12) weeks. There were no infections or wound complications at the time of last follow-up. Early temporising external fixation for the acute management of displaced calcaneus fractures is a safe and effective method to reduce and stabilise the foot and may decrease the time to definitive fixation. There were no complications related to the use of the external fixator in this series.


High-throughput transcriptome analysis reveals that the loss of Pten activates a novel NKX6-1/RASGRP1 regulatory module to rescue microphthalmia caused by Fgfr2-deficient lenses.
FGFR signaling is critical to development and disease pathogenesis, initiating phosphorylation-driven signaling cascades, notably the RAS-RAF-MEK-ERK and PI3 K-AKT cascades. PTEN antagonizes FGFR signaling by reducing AKT and ERK activation. Mouse lenses lacking FGFR2 exhibit microphakia and reduced ERK and AKT phosphorylation, widespread apoptosis, and defective lens fiber cell differentiation. In contrast, simultaneous deletion of both Fgfr2 and Pten restores ERK and AKT activation levels as well as lens size, cell survival and aspects of fiber cell differentiation; however, the molecular basis of this "rescue" remains undefined. We performed transcriptomic analysis by RNA sequencing of mouse lenses with conditional deletion of Fgfr2, Pten or both Fgfr2 and Pten, which reveal new molecular mechanisms that uncover how FGFR2 and PTEN signaling interact during development. The FGFR2-deficient lens transcriptome demonstrates overall loss of fiber cell identity with deregulated expression of 1448 genes. We find that ~ 60% of deregulated genes return to normal expression levels in lenses lacking both Fgfr2 and Pten. Further, application of customized filtering parameters to these RNA-seq data sets identified 68 high-priority candidate genes. Bioinformatics analyses showed that the cis-binding motif of a high-priority homeodomain transcription factor, NKX6-1, was present in the putative promoters of ~ 78% of these candidates. Finally, biochemical reporter assays demonstrate that NKX6-1 activated the expression of the high-priority candidate Rasgrp1, a RAS-activating protein. Together, these data define a novel regulatory module in which NKX6-1 directly activates Rasgrp1 expression to restore the balance of ERK and AKT activation, thus providing new insights into alternate regulation of FGFR downstream events.


New bioresorbable wraps based on oxidized polyvinyl alcohol and leukocyte-fibrin-platelet membrane to support peripheral nerve neurorrhaphy: preclinical comparison versus NeuraWrap.
Nerve wrapping improves neurorrhaphy outcomes in case of peripheral nerve injuries (PNIs). The aim of this preclinical study was to assess the efficacy of two novel biodegradable wraps made of a synthetic 1% oxidized polyvinyl alcohol (OxPVA) and a natural leukocyte-fibrin-platelet membrane (LFPm) versus the commercial product NeuraWrap. After rats sciatic nerve transection and neurorrhaphy, the wraps were implanted and compared for functional outcome, by sciatic function index assessment; structural characteristics, by histological/immunohistochemical analysis; ultrastructural features, by transmission electron microscopy. Moreover, a morphometric study was also performed and collagen distribution was observed by Second Harmonic Generation microscopy. After 12 weeks from implantation, all wraps assured nerve function recovery; no scar tissue/neuromas were visible at dissection. LFPm wraps were completely resorbed, while residues of OxPVA and NeuraWrap were observed. In all groups, biocompatibility was confirmed by the absence of significant inflammatory infiltrate. According to histological/immunohistochemical analysis and morphometric findings, OxPVA and LFPm wraps were both effective in preserving nerve integrity. These results assess that bioengineered OxPVA and LFPm wraps successfully guarantee favorable lesion recovery after PNI/neurorrhaphy and, in future, may be considered an interesting alternative to the commercial NeuraWrap.


Total full-thickness skin grafting for treating patients with extensive facial burn injury: A 10-year experience.
Facial burns are not only a severe burn injury, but result in psychological disturbance. The improvement of the methods of treating facial burns remains topical. The aim of the study was to evaluate the effectiveness of approach based on full-thickness skin autografting for facial burn injuries. During 2000-2019, ninety seven patients with the facial burn were treated in Burn Center. All patient were divided into two groups. The comparative analysis between groups was done. Group A was treated with full-thickness skin grafts (42 patients - 43.3%). Since 2010, total full-thickness skin graft was used in 11 patients from Group A. In group B, 55 patients (56.7%) were treated with split-thickness skin grafts, including 9 patients (16.4%) with total split-thickness skin graft transplantation. Total full-thickness skin graft was performed in case of a deep and extensive facial burn and cicatricial deformities. During the long-term period, a positive cosmetic result and the absence of indications for reconstructive operations were noted. The approach of facial burn treatment based on total full-thickness skin graft allows conditions for engraftment and adaptation of autograft, reduces the risk of scar developing and achieves maximum cosmetic results of treatment.


Corrigendum: Brucella-Induced Downregulation of lncRNA Gm28309 Triggers Macrophages Inflammatory Response Through the miR-3068-5p/NF-κB Pathway.
[This corrects the article DOI: 10.3389/fimmu.2020.581517.].


Challenges encountered in the analysis of phthalate esters in foodstuffs and other biological matrices.
Phthalate esters are ubiquitous environmental pollutants and are recognized as environmental endocrine disruptors because of their potential to elicit reproductive and developmental toxicity. Several phthalate esters have been listed by the US Environmental Protection Agency (EPA) as chemicals of concern. Determination of concentrations of phthalate esters in foodstuffs, typically present at sub to low nanogram-per-gram concentrations (between 0.1 and 100 ng g(-1)), is essential for assessment of human dietary exposure. However, phthalate esters are commonly present as contaminants in several laboratory products, including organic solvents, that are used in sample preparation and analysis. Therefore, accurate analysis of phthalates in food samples is a challenging task. In this review, we summarize the methods available for the determination of phthalate esters in foodstuffs and report on concentrations of phthalates in foodstuffs and potential sources of contamination by phthalates in the analysis of foodstuffs. We offer suggestions to eliminate and/or reduce background levels of contamination by phthalates in the analysis of food and other biological samples. We also introduce methods that are suitable for trace analysis of phthalates in a variety of liquid and solid food samples, in particular, a liquid-liquid extraction method for removal of lipids from food samples, because these can substantially reduce background levels of phthalates in the analytical procedure.


Professional Athletes Have Poorer Sleep Quality and Sleep Hygiene Compared With an Age-Matched Cohort.
To study sleep quality and sleep hygiene in professional athletes and an age-matched cohort. Cross-sectional study. Professional athletes and a sport medicine center. Professional rugby, netball and football athletes (n = 184) and attendees to a sport medicine center (n = 101). Participants completed an online survey. Sleep Hygiene Index (SHI) and Pittsburgh Sleep Quality Index (PSQI). Forty-five percent (n = 128) of respondents were aged between 18 and 24 years, 54% (n = 154) were men and 65% (n = 184) were professional athletes. The sleep duration of the professional athletes (mean rank 134.3, n = 181) was greater than the age-matched cohort (mean rank = 154.4, n = 101), U = 7835.0, z = -2.3, P = 0.02; however, they reported more sleep disturbance (mean ranking = 148.0, n = 181) than the age-matched cohort (mean rank = 129.8, n = 101), U = 7960.5 z = -2.5, P = 0.01, 2-tailed. Professional athletes had worse sleep regularity (mean rank = 152.3, n = 183) compared with the age-matched cohort (mean rank = 124.6, n = 101), U = 7448.5, z = -2.7, P = 0.006, 2-tailed, had an inferior sleep environment (mean rank = 149.5, n = 183) compared with the age-matched cohort (mean rank = 129.8, n = 101), U = 7959.5, z = -2.0, P = 0.047, and had more naps (mean rank = 156.2, n = 183) compared with the age-matched cohort (mean rank = 117.6, n = 101), U = 6729.0, z = -4.2, P = 0.00 0, 2-tailed. Professional athletes reported poorer sleep quality and sleep hygiene compared with an age-matched cohort, and difficulty falling asleep following competition. It is likely this is due to the stress of competition, training, and traveling. Because sleep plays an important role in postexercise recovery and has an impact on injury and athletic performance, it is important to have strategies to support better sleep quality and sleep hygiene in athletes.


Detect of Escherichia coli O157:H7 in ground beef samples using piezoelectric excited millimeter-sized cantilever (PEMC) sensors.
Piezoelectric-excited millimeter-sized cantilever (PEMC) sensors consisting of a piezoelectric and a borosilicate glass layer with a sensing area of 4 mm2 were fabricated. An antibody specific to Escherichia coli (anti-E. coli) O157:H7 was immobilized on PEMC sensors, and exposed to samples containing E. coli O157:H7 (EC) prepared in various matrices: (1) broth, broth plus raw ground beef, and broth plus sterile ground beef without inoculation of E. coli O157:H7 served as controls, (2) 100 mL of broth inoculated with 25 EC cells, (3) 100 mL of broth containing 25 g of raw ground beef and (4) 100 mL of broth with 25 g of sterile ground beef inoculated with 25 EC cells. The total resonant frequency change obtained for the broth plus EC samples were 16+/-2 Hz (n=2), 30 Hz (n=1), and 54+/-2 Hz (n=2) corresponding to 2, 4, and 6h growth at 37 degrees C, respectively. The response to the broth plus 25 g of sterile ground beef plus EC cells were 21+/-2 Hz (n=2), 37 Hz (n=1), and 70+/-2 Hz (n=2) corresponding to 2, 4, and 6 h, respectively. In all cases, the three different control samples yielded a frequency change of 0+/-2 Hz (n=6). The E. coli O157:H7 concentration in each broth and beef samples was determined by both plating and by pathogen modeling program. The results indicate that the PEMC sensor detects E. coli O157:H7 reliably at 50-100 cells/mL with a 3 mL sample.


Expression, purification, crystallization and structure determination of the N terminal domain of Fhb, a factor H binding protein from Streptococcus suis.
Fhb is a surface virulence protein from Streptococcus suis, which could aid bacterial evasion of host innate immune defense by recruiting complement regulator factor H to inactivate C3b deposited on bacterial surface in blood. Here we successfully expressed and purified the N terminal domain of Fhb (N-Fhb) and obtained crystals of the N-Fhb by sitting-drop vapor diffusion method with a resolution of 1.50 Å. The crystals belong to space group C2 with unit cell parameters a = 127.1 Å, b = 77.3 Å, c = 131.6 Å, α = 90°, β = 115.9°, γ = 90°. The structure of N-Fhb was determined by SAD method and the core structure of N-Fhb is a β sandwich. We speculated that binding of Fhb to human factor H may be mainly mediated by surface amino acids with negative charges.


Development of analytical method for simultaneous determination of five rodent unique bile acids in rat plasma using ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry.
Bile acids (BAs) are crucial for the diagnosis, follow-up, and prognostics of liver injuries and other BA metabolism related diseases. In particular, rodent unique BAs, α-muricholic acid (α-MCA), β-MCA, ω-MCA, tauro-α-MCA (α-TMCA), and β-TMCA, are valuable biomarkers for preclinical drug development. To the best of our knowledge, however, a simple, selective, sensitive, and robust analytical method for ω-MCA and taurine-conjugated MCAs has never been reported. We have developed a simple, selective, and sensitive analytical method for measurement of 16 BAs including the five rodent unique BAs in rat plasma using an ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-TOF-MS) method. Activated charcoal was utilized to prepare BA-free plasma, which served as the surrogate matrix for the preparation of calibration standards and quality control (QC) samples. Results of matrix effects evaluation suggested that the BA-free plasma could be adequate as a surrogate matrix for BAs determination. Three stable isotope labelled internal standards were separated by reverse phase UPLC using gradient elution and were detected by TOF-MS in negative ion mode. The calibration curve was linear for all BAs over a range of 10-25ng/mL to 1000-10,000ng/mL, with overall imprecision below 15% and 20% at lower limit of quantification (LLOQ), respectively. This analytical method was used to determine BA concentrations in more than 300 plasma samples from rats with liver injuries induced using α-naphthylisocyanate, carbon tetrachloride, or flutamide. The alteration of BA concentrations was most evident for necrosis, and cholestasis hepatotoxins, with more subtle effects by steatosis and idiosyncratic hepatotoxins. In conclusion, we have developed a simple, selective, and sensitive analytical method to measure plasma 16 BAs including 5 rodent unique BAs, α-MCA, β-MCA, ω-MCA, α-TMCA, and β-TMCA. Our data suggested that α-TMCA and β-TMCA could be useful for identification or prediction of liver injuries, a currently unmet need in preclinical toxicity. Our method using TOF-MS is useful to determine BAs in rat plasma and of use in structural analyses of metabolites in early stage of drug development.


Improved Microvascular Decompression in Treating Trigeminal Neuralgia: Application of Nest-Shaped Teflon Fibers.
To develop an improved technique for microvascular decompression (MVD) surgery using nest-shaped Teflon fibers. Eighteen consecutive patients with trigeminal neuralgia (TN) who underwent MVD using nest-shaped Teflon fibers between January 2012 and December 2013 were included in this investigation. During the surgery, the Teflon prosthesis was formed into a nest shape by gently pushing it from the center toward the periphery. Immediate postoperative outcomes were evaluated using a numerical rating scale score, and patients were followed up for recurrence. Immediately after the surgery, pain was completely relieved in 16 patients (88.9%) and partially relieved in 2 patients (11.1%). Seven patients (38.9%) developed postoperative complications. All complications were successfully mitigated before discharge. The patients were followed up for 2.0-3.4 years. During follow-up, recurrence was observed in 2 patients (11.1%). No Teflon adhesion or Teflon-induced granuloma was found. The nest-shaped Teflon fibers in MVD surgery for TN is safe and applicable. The long-term outcomes and the comparison between hollow nest-shaped implants and the standard cigar-shaped implants should be assessed in future investigations with larger sample sizes.


Phosphorylation of smooth muscle myosin light chain kinase by the catalytic subunit of adenosine 3': 5'-monophosphate-dependent protein kinase.
Turkey gizzard smooth muscle light chain kinase was purified by affinity chromatography on calcium dependent regulator weight of 125,000 +/- 5,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When myosin light chain kinase is incubated with the catalytic subunit of cyclic AMP-dependent protein kinase, 1 mol of phosphate is incorporated per mol of myosin kinase. Brief tryptic digestion of the 32P-labeled myosin kinase liberates a single radioactive peptide with a molecular weight of approximately 22,000. Phosphorylation of myosin kinase results in a 2-fold decrease in the rate at which the enzyme phosphorylates the 20,000-dalton light chain of smooth muscle myosin. These results suggest that cyclic AMP has a direct effect on actin-myosin interaction in smooth muscle.


Apomorphine-induced behaviour during the oestrous cycle of the rat.
The effect of various doses of apomorphine (APO) (25, 250, 400 and 750 micrograms/kg, s.c.) on open field behaviour, stereotyped behaviour, body temperature and concentrations of serum oestradiol was studied in cycling females and in ovariectomized rats. With the exception of grooming, the hormonal variations during the cycle, or the ovariectomy, did not have an effect on behaviour related to stimulation of presynaptic dopamine (DA) receptors. The endocrine status on proestrus (PE), characterized by an increase in serum oestradiol, did influence hyperlocomotion and hypothermia induced by apomorphine; the former being attenuated and the latter increased, as compared to the other phases of the cycle. Ovariectomy resulted in an increase in the stimulatory effect of apomorphine on locomotion. Stereotypy induced by apomorphine was unaltered by hormonal variations during the cycle and it was slightly attenuated by removal of the ovaries. During phases of low levels of oestrogen (oestrus, metestrus) apomorphine significantly increased the levels of serum oestradiol, determined 30 min after the administration of drug. It is concluded that the various DAergic mechanisms in brain are differentially affected by hormonal variations during the cycle and by ovariectomy.


Some effects of prophylactic aspirins and heparins on concurrent arterial and venous thrombosis in the same animal.
The antithrombotic effect of unfractionated heparin, a low-molecular weight heparin, oral acetylsalicylic acid, intravenous lysine-aspirin and oral soluble aspirin was measured on platinum wire simultaneously in artery and vein, in groups of 12-20 rats with concurrent untreated controls. Subcutaneous low molecular weight heparin reduced mean thrombus weight by 26% in the artery only. Venous thrombosis was reduced 56% by oral soluble acetylsalicylic acid and 27% by oral aspirin. Other reductions did not reach statistical significance. While thrombus deposition was induced by both 2-cm (upper site) and 1-cm (lower site) wires, the results obtained on the 2-cm wires in the upper vein and artery were more reliable [corrected].


Effect of repeated reproductive cycles on maternal nutritional status, lactational performance and litter growth in ad libitum-fed and chronically food-restricted rats.
To evaluate the combined effects of repeated, closely spaced reproductive cycles and dietary intake on maternal nutritional status, lactational performance and litter growth, rats were fed ad libitum or 75 or 60% of ad libitum intake. Dietary treatment began 28 d before breeding and continued until d 14 of the first (L1) or second (L2) lactation. Body weight and carcass fat concentration of dams and their litters were affected in the 75% group; milk yield, milk protein and lactose concentrations and energy content were affected only in the 60% group. Dams and their litters were heavier, had more total carcass protein and higher plasma albumin values in L2 than L1. There was no effect of reproductive period on milk yield or composition. These results indicate that repeated reproductive cycles did not compromise maternal nutritional status, lactational performance or litter growth. Improved outcomes in L2 among the restricted rats appeared to result from gains during the interval between reproductive periods and early in the second pregnancy.


The Saint-Joseph Acetabular score: a reproducible and accurate prediction of the outcome of open reduction and internal fixation of acetabular fractures.
Acetabular fractures are associated to an increased risk of subsequent hip osteoarthritis. The only available score for post-operative reduction assessment in acetabular fractures (Matta's score) is x-ray based. CT-scan was shown superior to X-rays in post-operative reduction assessment. We aim to describe a CT-scan-based post-operative reduction score in acetabular fractures and evaluate its accuracy and reproducibility. This is a retrospective study that includes 138 patients who underwent surgery for an acetabular fracture in our tertiary referral centre with a mean follow-up of 104.39±42.2 months. The post-operative CT-scan was reviewed and residual displacement (maximum gap and step) measured by three independent observers. The association between the occurrence of THA and the CT-scan measurements was evaluated. This led to a new prognostic score. The interobserver reliability and accuracy of this score were calculated. Interobserver reproducibility for the residual maximal gap was 0.82 (95% CI [0.70-0.89]) and 0.61 (95% CI [0.52-0.70]) for the residual maximal step displacement measurements. We created a score from a logistic regression model, attributing 1 point for every 1 mm of residual maximal step displacement and 1 point for every 2 mm of residual maximal gap displacement. The interobserver reproducibility of this score was 0.78 (95% CI [0.71-0.84]), and its AUC was 0.79 (95% CI [0.69-0.88]). This is the first CT-scan-based score for the assessment of residual displacement of a surgically treated acetabular fracture. It shows good interobserver reproducibility and accuracy in predicting the risk for secondary THA. It should be regularly used per-operatively (if per-operative 3D imaging is available) and post-operatively to predict the prognosis.


Continuous cardiac output monitoring with pulse contour analysis: a comparison with lithium indicator dilution cardiac output measurement.
Pulse contour analysis can be used to provide beat-to-beat cardiac output (CO) measurement. The current study sought to evaluate this technique by comparing its results with lithium dilution CO (LiCO) measurements. Prospective, observational study. Surgical intensive care unit. Twenty-two patients after cardiac or major noncardiac surgery. After initial calibration of the pulse contour CO (PCO) method, CO was measured by PCO and by LiCO methods at 4, 8, 16, and 24 hrs. Recalibration of PCO was performed every 8 hrs. The systemic vascular resistance and dynamic response characteristics of the arterial catheter-transducer system were measured at each time point to determine whether these influenced the agreement between PCO and LiCO methods. There was an excellent correlation between methods (r = .94). Bias was small (-0.005 L/min), and clinically acceptable limits of agreement were demonstrated between techniques. Although many catheter-transducer systems had poor dynamic response characteristics, this did not influence the level of agreement between the two techniques. An increase in systemic vascular resistance between two time points did tend to cause overestimation of LiCO by the PCO. PCO measurement compared well with the lithium dilution method and can be considered an accurate technique for measuring beat-to-beat CO with limited risk to the patient.