Effect of ACTH on glucose uptake by hepatic cells and its effect on hepatic enzymes in presence of insulin and acetylcholine.
An intrinsic hypoglycaemic activity has been attributed to ACTH. This action of ACTH is essentially mediated by increased insulin secretion. In the present study, the direct action of ACTH on glucose uptake by pigeon hepatocytes has been studied by in vitro technique. The results have shown that ACTH has a direct influence on glucose uptake and this action is not additive in presence of insulin. Glucose uptake in presence ACTH with Acetylcholine was not any more than what was obtained with ACTH alone. These observations have been taken to indicate a non-existence of synergistic action of ACTH with insulin or acetylcholine in promoting glucose uptake by avian liver cells.


Critical cerebral blood flow thresholds studied by SPECT using xenon-133 and iodine-123 iodoamphetamine.
Regional cerebral blood flow (rCBF) was assessed in patients with cerebrovascular diseases (CVD) by single photon emission computed tomography (SPECT) using 133Xe and N-isopropyl-p-[123I]iodoamphetamine ([123I]IMP). The purpose of this study was to determine the infarcted and symptomatic blood flow thresholds of cerebral cortex by SPECT. In the 133Xe inhalation method rCBF was calculated by employing the Celsis modification of the Kanno and Lassen algorithm. In the [123I]IMP SPECT quantitation was obtained by counting-rate ratio for the low flow lesion to the contralateral homologous region which was presumed to be normal (Lesion/Normal ratio, L/N ratio). The infarcted and symptomatic blood flow thresholds by 133Xe SPECT was 19-23 and 33-36 ml/100 g/min, respectively. While, those of L/N ratio in the [123I]IMP SPECT were 39-48 and 65-72%, respectively. There was a significant correlation between the ischemic degrees evaluated by 133Xe and [123I]IMP SPECT studies.


Suppression of DELLA signaling induces procambial cell formation in culture.
The post-embryonic growth of plants requires the activities of apical meristems and lateral meristems. In the meristems, self-proliferation and differentiation of stem cells is tightly modulated by plant hormone signaling networks and specific transcription factors. Despite extensive studies on stem cell maintenance in plants, the mechanism by which stem cells are initially established is largely unknown. Vascular stem cells consisting of procambial/cambial cells give rise to xylem and phloem cells. In this study, we analyzed the establishment of procambial cells using the in vitro culture system VISUAL, in which mesophyll cells rapidly differentiate into xylem tracheary elements and phloem sieve elements via procambial cells. We found that procambial cell formation in VISUAL is initiated by light, which can be replaced by application of gibberellin (GA). Gibberellin was able to promote procambial cell formation through degradation of DELLA, whereas light did not elevate the endogenous GA content. Indeed, light in combination with bikinin reduced the accumulation of DELLA protein in VISUAL. Consistently, overexpression of a constitutively active DELLA protein repressed vascular cell differentiation even under light. These combined results suggest that DELLA signaling suppresses procambial cell formation during vascular development in VISUAL.


p23 protects the human aryl hydrocarbon receptor from degradation via a heat shock protein 90-independent mechanism.
The aryl hydrocarbon receptor (AHR) is a ligand-activated signaling molecule which is involved in diverse biological functions ranging from cancer metastasis to immune regulation. This receptor forms a cytoplasmic complex with Hsp90, p23, and XAP2. We have previously reported that down-regulation of p23 triggers degradation of the AHR protein, uncovering a potentially dynamic event which controls the cellular AHR levels without ligand treatment. Here we investigate the underlying mechanisms for this p23 effect using wild-type HeLa and the p23 knockdown HeLa cells. Reduction of the Hsp90 and XAP2 contents, however, did not affect the AHR protein levels, implying that this p23 effect on AHR is more than just alteration of the cytoplasmic complex dynamics. Association of p23 with Hsp90 is not important for the modulation of the AHR levels since exogenous expression of p23 mutants with modest Hsp90-binding affinity effectively restored the AHR message and protein levels. The protein folding property of p23 which resides at the terminal 50-amino acid region is not involved for this p23 effect. Results from our interaction study using the affinity purified thioredoxin fusion proteins and GST fusion proteins showed that p23 directly interacts with AHR and the interaction surface lies within AHR amino acid 1-216 and p23 amino acid 1-110. Down-regulation of the p23 protein content promotes the ubiquitination of AHR, indicating that p23 protects AHR from the ubiquitin-meditated protein degradation.


A comparison of neuraxial block versus general anesthesia for elective total hip replacement: a meta-analysis.
A recent meta-analysis showed that compared with general anesthesia (GA), neuraxial block reduced many serious complications in patients undergoing various types of surgeries. It is not known whether this finding from studying heterogeneous patient groups is applicable to a particular surgical patient population. We performed the present meta-analysis to determine whether anesthesia choice affected the outcome after elective total hip replacement (THR). Medline (1966 to August 2005), MD Consult (1966 to August 2005), BIOSIS (1969 to August 2005), and EMBASE (1969 to August 2005) databases were searched. Randomized and quasi randomized studies comparing GA and neuraxial (spinal or epidural) block for elective THR were included in this analysis. Ten independent trials, involving 330 patients under GA and 348 patients under neuraxial block, were identified and analyzed. Pooled results from five trials showed that neuraxial block significantly decreased the incidence of radiographically diagnosed deep venous thrombosis or pulmonary embolism. The odds ratio (OR) for deep venous thrombosis was 0.27 with 95% confidence interval (CI) 0.17-0.42. The OR for pulmonary embolism was 0.26 with 95% CI 0.12-0.56. Neuraxial block also decreased the operative time by 7.1 min/case (95% CI 2.3-11.9 min) and intraoperative blood loss by 275 mL/case (95% CI 180-371 mL). Data from three trials showed that patients under neuraxial block for THR were less likely to require blood transfusion than were patients under GA (21/177 = 12% vs 62/188 = 33% of patients transfused, P < 0.001 by z-test). The OR for this comparison was 0.26. However, the CIs were wide and compatible with both no effect and a nine-tenths reduction (95% CI 0.06-1.05). Patients undergoing elective THR under neuraxial anesthesia seem to have better outcomes than those under GA.


Dynamic Changes of Metabolic Syndrome Alter the Risks of Cardiovascular Diseases and All-Cause Mortality: Evidence From a Prospective Cohort Study.
Background: Metabolic syndrome (MetS) at baseline increases the risks of cardiovascular diseases (CVD) and all-cause mortality. However, MetS status is changeable during follow-up. The associations of dynamic changes of MetS with CVD and all-cause mortality remain unclear. Methods: Thirty-one thousand four hundred eighty-one eligible subjects were included from the Kailuan cohort. Dynamic changes of MetS were divided into four patterns as MetS-free, MetS-developed, MetS-recovery and MetS-stable. The outcomes were CVD, all-cause mortality, and the subtypes of CVD as myocardial infarction (MI), stroke and heart failure. Multiple Cox regression models were used to calculate the adjusted hazard ratios (HRs) and confidence intervals (95% CIs). Results: Altered risks of CVD, the subtypes of CVD, and all-cause mortality were observed among different dynamic patterns of MetS. Compared with the MetS-free group, MetS-developed group increased the risks of CVD (HR = 1.78, 95% CI = 1.51-2.11), MI (HR = 1.54, 95% CI = 1.01-2.34), stroke (HR = 1.78, 95% CI = 1.45-2.18), and heart failure (HR = 1.63, 95% CI = 1.11-2.39). MetS-recovery group decreased these risks with the HRs of 0.59 (95% CI = 0.48-0.72) for CVD, 0.62 (95% CI = 0.41-0.96) for MI, 0.59 (95% CI = 0.46-0.75) for stroke, and 0.56 (95% CI = 0.34-0.91) for heart failure compared with the MetS-stable group. However, the increased risk in the MetS-developed group and the decreased risk in the MetS-recovery group were not significant for all-cause mortality. When stratified by the onset age of MetS status change, early development of MetS (<50 years) had higher risks of CVD (HR = 2.20, 95% CI = 1.58-3.05), MI (HR = 2.35, 95% CI = 1.00-5.50), stroke (HR = 2.05, 95% CI = 1.38-3.05), heart failure (HR = 2.63, 95% CI = 1.15-6.04), and all-cause mortality (HR = 1.61, 95% CI = 1.13-2.30) than late development (≥50 years). Early recovery of MetS had lower risks with the HRs of 0.38 (95% CI = 0.24-0.59) for CVD, 0.43 (95% CI = 0.18-1.06) for MI, 0.37 (95% CI = 0.21-0.64) for stroke, 0.30 (95% CI = 0.09-1.04) for heart failure, and 0.68 (95% CI = 0.43-1.06) for all-cause mortality than late recovery. Conclusion: Dynamic changes of MetS altered the risks of CVD and all-cause mortality, especially in individuals with an early onset age. These findings highlight the importance of dynamic changes of MetS and onset age on the prevention and control for CVD.


Impact of Gender Disparities on Short-Term and Long-Term Patient Reported Outcomes and Satisfaction Measures After Elective Lumbar Spine Surgery: A Single Institutional Study of 384 Patients.
There is a paucity of data determining the impact that gender disparities have on spine outcomes, particularly perception of health and satisfaction. The aim of this study was to determine whether there is a difference in 3-month and 1-year patient-reported outcomes and satisfaction after elective lumbar spine surgery. This was a retrospectively analyzed study from a maintained prospective database of 384 patients who underwent elective lumbar spine surgery. Patients were categorized by gender (men, n = 199; women, n = 185). Patient-reported outcome instruments (Oswestry disability index, visual analogue scale-back pain/leg pain, EuroQol visual analogue scale, and EuroQol 5 dimensions questionnaire) were completed before surgery, then at 3 and 12 months after surgery along with patient satisfaction measures. Baseline patient demographics, comorbidities, and operative variables were similar between both cohorts. The female cohort had a slightly longer hospital stay than male cohort (P = 0.007). Baseline patient-reported outcome measures were different between both cohorts, with female patients having more Oswestry disability index (23.8 vs. 20.4; P ≤ 0.0001) and visual analogue scale-back pain (7.2 vs. 6.2; P = 0.0004), and a lower EuroQol 5 dimensions questionnaire (0.34 vs. 0.49; P = 0.0001) compared with the male cohort. At 1-year follow-up, the male cohort had a significantly more mean change in visual analogue scale-leg pain (-3.9 vs. -2.8; P = 0.04) and trended to have more mean change in visual analogue scale-back pain (-3.4 vs. -2.5; P = 0.06) and EuroQol visual analogue scale (8.6 vs. 3.4; P = 0.054) scores compared with the female cohort. At 1-year a significantly more portion in the male cohort found that surgery met their expectations compared with the female cohort (65.0% vs. 49.5%; P = 0.02). Our study suggests that there may be differences in perception of health, pain, and disability between men and women at baseline, short-term and long-term follow-up that may influence overall patient satisfaction.


A comparison of dentine removal using safety or conventional Hedstrom files.
Mandibular molars (n = 20) were divided and the mesial roots mounted in an acrylic mould which allowed them to be removed, sectioned horizontally and then reassembled. The furcal root canal wall thickness 7 mm from the apex was measured prior to instrumentation. The roots were allocated to two groups (n = 10). One group was instrumented using the step back technique with safety Hedstroms in one canal and conventional Hedstrom files in the other canal. The second group was instrumented using a crown down approach (using files and Gates Glidden drills to provide early radicular access). The furcal root canal wall thickness was then remeasured and the amount of dentine removed calculated. The results showed that the safety Hedstrom file removed less dentine from the furcal region of the canal compared to the conventional Hedstrom file. This however was not statistically significant (Paired t test, n = 10, p > 0.05). The amount of dentine removed using the crown down technique was significantly greater than that using the step back technique (Two sample t, n = 20, p < 0.01). In conclusion, less dentine was removed from the furcal region with safety Hedstrom files compared to conventional Hedstrom files; however, Gates Glidden drills removed more dentine than either type of hard file.


Parity violation energy of biomolecules--I: polypeptides.
A computational algorithm is developed to compute the energy of parity-violation (E(pv)) due to weak-nuclear forces acting within chain polymers. The method is applied to estimate the magnitude of E(pv) associated with the folding of alpha-helices in polypeptide chains and, in turn, of typical proteins. Implications to the development of biological homochirality in nature are discussed in terms of a simple evolutionary model to assess the likelihood for energies of the order of the computed E(pv) to have determined the direction of natural homochirality.


Nutrition in the elderly: nutritional aspects of chronic diseases.
Osteoporosis, diseases of the oral cavity, GI disorders, obesity, diabetes, cancer, and heart disease are highly influenced by dietary factors. Older patients may benefit from calcium supplementation even if the osteoporotic process is well under way. Certain GI disorders respond to increased fiber intake. Obesity and diabetes are sensitive to dietary changes, and diabetes sometimes can be prevented or reversed in its early stages with diet and exercise. High fruit and vegetable consumption seems to lower cancer risk. No strong evidence suggests that nutritional supplements improve cognition.


Erratum: Activation and Conjugation of Soluble Polysaccharides using 1-Cyano-4-Dimethylaminopyridine Tetrafluoroborate (CDAP).
An erratum was issued for: Activation and Conjugation of Soluble Polysaccharides using 1-Cyano-4-Dimethylaminopyridine Tetrafluoroborate (CDAP). A figure was updated. Figure 4 was updated from: Figure 4: Representative results for CDAP activation of dextran. Typical standard curves for the (A) resorcinol/sulfuric acid and (B) TNBS assays. The assay results for dextran activated with 0.25 and 0.5 mg CDAP/mg dextran are shown. Glucose was used as the standard for the resorcinol assay. Dextran, in mg/mL, is divided by 100 kDa to give a molar concentration. The hydrazide concentration is determined using ADH as the standard and the results expressed as µM Hz. (C) Calculation of hydrazide: dextran ratios.The level of derivatization was calculated as hydrazides per 100 kDa of dextran to facilitate the comparison between polymers of different average molecular weights. The % weight ratio of g ADH/g dextran was calculated using a MW of 174 g/mole for ADH. Please click here to view a larger version of this figure. to: Figure 4: Representative results for CDAP activation of dextran. Typical standard curves for the (A) resorcinol/sulfuric acid and (B) TNBS assays. The assay results for dextran activated with 0.25 and 0.5 mg CDAP/mg dextran are shown. Glucose was used as the standard for the resorcinol assay. Dextran, in mg/mL, is divided by 100 kDa to give a molar concentration. The hydrazide concentration is determined using ADH as the standard and the results expressed as µM Hz. (C) Calculation of hydrazide: dextran ratios.The level of derivatization was calculated as hydrazides per 100 kDa of dextran to facilitate the comparison between polymers of different average molecular weights. The % weight ratio of g ADH/g dextran was calculated using a MW of 174 g/mole for ADH. Please click here to view a larger version of this figure.


Aggregation-induced Emission Properties of Triphenylamine Chalcone Compounds.
Two triphenylamine chalcone derivatives 1 and 2 were synthesized through the Vilsmeier-Haack reaction and Claisen-Schmidt condensation reaction. Through ultraviolet absorption spectroscopy and fluorescence emission spectroscopy experiments, it was confirmed that these two compounds exhibited good aggregation-induced emission (AIE) behavior in ethanol/water mixtures. The solvent effect test showed with the increase of the orientation polarizability of the solvent, the Stokes shift in the solvent of compound 1 and compound 2 shows a linear change trend. Through solid state fluorescence test and universal density function theory (DFT), the existence of π-π stacking interaction in the solid state of the compound has been studied, resulting in weak fluorescence emission. pH has no effect on the fluorescence intensity of the aggregate state of excited state intramolecular proton transfer (ESIPT) molecules in an acidic environment, but greatly weakens its fluorescence intensity in an alkaline environment. Cyclic voltammetry (CV) test shows that compound 1 was more prone to oxidation reaction than compound 2. The results of thermal stability test show that the thermal stability of compound 1 was better than that of compound 2, indicating that triphenylamine chalcone derivatives can improve the thermal stability of compounds by increasing the number of branches.


Targeting transcription factors for therapeutic benefit in rheumatoid arthritis.
Rheumatoid arthritis (RA) is a destructive inflammatory autoimmune disease that causes pain and disability. Many of the currently available drugs for treating RA patients are aimed at halting the progression of the disease and alleviating inflammation. Further, some of these treatment options have drawbacks, including disease recurrence and adverse effects due to long-term use. These inefficiencies have created a need for a different approach to treating RA. Recently, the focus has shifted to direct targeting of transcription factors (TFs), as they play a vital role in the pathogenesis of RA, activating key cytokines, chemokines, adhesion molecules, and enzymes. In light of this, synthetic drugs and natural compounds are being explored to target key TFs or their signaling pathways in RA. This review discusses the role of four key TFs in inflammation, namely NF-κB, STATs, AP-1 and IRFs, and their potential for being targeted to treat RA.


A Micro-agar Salt Bridge Electrode for Analyzing the Proton Turnover Rate of Recombinant Membrane Proteins.
To date, more than 50% of all pharmacological drugs target the transport kinetics of membrane proteins. The electrophysiological characterization of membrane carrier proteins reconstituted in lipid bilayer membranes is a powerful but delicate method for the assessment of their physicochemical and pharmacological properties. The substrate turnover number is a unique parameter that allows the comparison of the activity of different membrane proteins. In an electrogenic transport, the gradient of the translocated substrate creates a membrane potential that directly correlates to the substrate turnover rate of the protein. By using silver chloride electrodes, a diffusion potential, also called liquid junction potential, is induced, which alters electrode potential and significantly disturbs precise membrane potential measurements. Diffusion potential can be minimized by a salt bridge, which balances electrode potential. In this article, a micro-agar salt bridge is designed to improve the electrophysiological set-up, which uses micropipettes for the membrane formation. The salt solution is filled into a microcapillary pipette tip, stabilized by the addition of agarose, and can be easily mounted to a standard electrode. The electrode potential of a micro-salt bridge electrode is more stable compared to a standard electrode. The implementation of this system stabilizes electrode potential and allows more precise measurements of membrane potential generated by a pH gradient. Using this system, the proton turnover rates of the mitochondrial carriers UCP1 and UCP3 are reinvestigated and compared to earlier measurements.


Subtyping of Human Papillomavirus-Positive Cervical Cancers Based on the Expression Profiles of 50 Genes.
Human papillomavirus-positive (HPV+) cervical cancers are highly heterogeneous in molecular and clinical features. However, the molecular classification of HPV+ cervical cancers remains insufficiently unexplored. Based on the expression profiles of 50 genes having the largest expression variations across the HPV+ cervical cancers in the TCGA-CESC dataset, we hierarchically clustered HPV+ cervical cancers to identify new subtypes. We further characterized molecular, phenotypic, and clinical features of these subtypes. We identified two subtypes of HPV+ cervical cancers, namely HPV+G1 and HPV+G2. We demonstrated that this classification method was reproducible in two validation sets. Compared to HPV+G2, HPV+G1 displayed significantly higher immune infiltration level and stromal content, lower tumor purity, lower stemness scores and intratumor heterogeneity (ITH) scores, higher level of genomic instability, lower DNA methylation level, as well as better disease-free survival prognosis. The multivariate survival analysis suggests that the disease-free survival difference between both subtypes is independent of confounding variables, such as immune signature, stemness, and ITH. Pathway and gene ontology analysis confirmed the more active tumor immune microenvironment in HPV+G1 versus HPV+G2. HPV+ cervical cancers can be classified into two subtypes based on the expression profiles of the 50 genes with the largest expression variations across the HPV+ cervical cancers. Both subtypes have significantly different molecular, phenotypic, and clinical features. This new subtyping method captures the comprehensive heterogeneity in molecular and clinical characteristics of HPV+ cervical cancers and provides potential clinical implications for the diagnosis and treatment of this disease.


What's Right With Men? Gender Role Socialization and Men's Positive Functioning.
This study explored relations between conformity to masculine norms, gender role conflict, hope, and psychological well-being among a sample of 389 men from a university, with a predominantly White student body, located in the Midwestern United States. Bivariate correlations revealed that men's conformity to masculine norms and gender role conflict were positively correlated. Bivariate correlations revealed no significant relations between conformity to masculine norms, trait hope, and psychological well-being. Gender role conflict was associated with decreased hope and psychological well-being. Results of path analysis explained relations between conformity to masculine norms, gender role conflict, trait hope, and psychological well-being. This indicates that gender role conflict may contribute to lower trait hope and psychological well-being for college men. Although several aspects of conformity to masculine norms had positive associations with hope, these relations were significant and negative when men experienced gender role conflict. This work fills an important gap in the literature by examining the unique relations of conformity to masculine norms and gender role conflict to men's positive functioning. Results are discussed within the context of positive psychological theories including Frederickson's broaden and build theory of positive emotions, hope theory, Ryff's model of psychological well-being, and self-determination theory. Implications and future directions are discussed.


Patterns of suicide and other trespassing fatalities on state-owned railways in greater Stockholm; implications for prevention.
Each year, approximately 80-100 people are killed on state-owned railways due to train-person collisions in Sweden. Underlying causes are suicide and accidents; suicide constituting a vast majority. Earlier Swedish studies at a national level revealed a relation between population density and incident frequency, however, with places of occurrence often located to the outskirts of cities some distance away from station areas where victims can await approaching trains in seclusion. The aim of this study was to investigate whether this national pattern also applies to larger urban areas such as greater Stockholm, and to discuss preventative implications based on these observations. All registered incidents (N = 41) where people were hit or run-over by trains with a fatal outcome over the four-year period 2005-2008 were investigated. Results deviating from the national pattern include that most incidents occur at station areas, and that most victims enter the tracks from platforms. Passing express trains appear to be overrepresented, compared to commuter trains. Due to a low number of cases, our observations must be interpreted with caution. However, they imply that preventative measures in this type of area should focus on platform safety foremost, especially protection against rapid trains passing by station areas.


Classification of Multiclass Histopathological Breast Images Using Residual Deep Learning.
Pathologists need a lot of clinical experience and time to do the histopathological investigation. AI may play a significant role in supporting pathologists and resulting in more accurate and efficient histopathological diagnoses. Breast cancer is one of the most diagnosed cancers in women worldwide. Breast cancer may be detected and diagnosed using imaging methods such as histopathological images. Since various tissues make up the breast, there is a wide range of textural intensity, making abnormality detection difficult. As a result, there is an urgent need to improve computer-assisted systems (CAD) that can serve as a second opinion for radiologists when they use medical images. A self-training learning method employing deep learning neural network with residual learning is proposed to overcome the issue of needing a large number of labeled images to train deep learning models in breast cancer histopathology image classification. The suggested model is built from scratch and trained.


A method for preparing collagen graft materials.
The aim of this research was to develop a method of local production of collagen graft materials which are presently imported. The following methods were used to produce collagen membrane and sponge from human placentas and rat tail tendons. Collagen type I was isolated from human placenta and rat tail tendon by acetic acid extraction and characterised by SDS-PAGE. The collagen sponge was prepared by dissolving the collagen in HCl. The resulting dispersion was poured into a glass container, freeze-dried and then cross-linked by immersion in glutaraldehyde solution. It was then washed with distilled water and freeze-dried again. The collagen membrane was also similarly prepared by dispersing lyophilized collagen in HCl but then mixed with glutaraldehyde, exposed to U.V. light and later air dried.


Radiotherapy and concomitant temozolomide may improve survival of elderly patients with glioblastoma.
Survival of elderly patients with glioblastoma (GBM) is poor, but improves with tumor resection and radiotherapy (RT). Concurrent temozolomide (TMZ) chemotherapy during RT improves the survival of younger patients with GBM, but the benefit in elderly patients is unclear. Medical records of patients ≥65 years old with primary GBM, histologically confirmed at Memorial Sloan-Kettering Cancer Center and treated with RT, were reviewed. Survival was associated with patient (age, performance status), tumor (single or multiple), and treatment (extent of surgery, RT field, technique, fractionation and use of concurrent TMZ) characteristics in a multivariable Cox regression model. Grade ≥3 hematologic toxicity rates were compared to reported rates in younger patients. Median age of the 291 patients studied was 71 years. Longer survival was associated with younger age, tumor resection, and concomitant TMZ and RT (p < 0.01). Concurrent TMZ and RT improved median survival of patients with favorable prognostic factors from 12 to 21 months and from 10 to 13 months in patients 65-70 and ≥71 years old, respectively. Concomitant TMZ and RT increased the 2 year OS rate from 14 to 41 % and from 5 to 24 % in patients 65-70 and ≥71 years old, respectively. Grade 3-4 thrombocytopenia was significantly more frequent in the present cohort. Survival of elderly patients with GBM may be prolonged with the use of concomitant TMZ during RT. An ongoing randomized study will determine the benefit of this approach in a prospective fashion.


Adaptation of flocculent anammox culture to low temperature by cold shock: long-term response of the microbial population.
Partial nitritation-anammox (PN/A) process will substantially reduce the costs for the removal of nitrogen in the mainstream of municipal sewage. However, one of the mainstream PN/A challenges is to reduce the time necessary for the adaptation of anammox bacteria to lower temperatures in mild climates. In this study, we exposed anammox flocculent culture to cold shocks [35°C → 5°C (8 h) → 15°C] and evaluated long-term cold shock response. Over a post-shock period of 40 d at 15°C, the nitrogen removal rates in the shocked culture were significantly higher compared to control, with maximum rates up to 0.082 and 0.033 kg-N/kg-VSS/d or 0.164 and 0.076 kg-N/m3/d, for shocked culture and control, respectively. In the corresponding semi-batch cycles, the shocked culture was on average 136 ± 101% more active than the control, due to the negative effect of cold shock on side populations and more active anammox cells. Per FISH, Ca. Brocadia anammoxidans and Ca. Scalindua survived the shock and remained present throughout. Thus, both anammox microorganisms seem to respond favourably to cold shocks. In sum, we provide further evidence that cold shocks accelerate the adaptation of anammox to the mainstream of municipal WWTPs. Further, for the first time, we report the long-term adaptive response of anammox to cold shocks.


The Language of Glove: Wireless gesture decoder with low-power and stretchable hybrid electronics.
This communication describes a glove capable of wirelessly translating the American Sign Language (ASL) alphabet into text displayable on a computer or smartphone. The key components of the device are strain sensors comprising a piezoresistive composite of carbon particles embedded in a fluoroelastomer. These sensors are integrated with a wearable electronic module consisting of digitizers, a microcontroller, and a Bluetooth radio. Finite-element analysis predicts a peak strain on the sensors of 5% when the knuckles are fully bent. Fatigue studies suggest that the sensors successfully detect the articulation of the knuckles even when bent to their maximal degree 1,000 times. In concert with an accelerometer and pressure sensors, the glove is able to translate all 26 letters of the ASL alphabet. Lastly, data taken from the glove are used to control a virtual hand; this application suggests new ways in which stretchable and wearable electronics can enable humans to interface with virtual environments. Critically, this system was constructed of components costing less than $100 and did not require chemical synthesis or access to a cleanroom. It can thus be used as a test bed for materials scientists to evaluate the performance of new materials and flexible and stretchable hybrid electronics.


An efficient churn prediction model using gradient boosting machine and metaheuristic optimization.
Customer churn remains a critical challenge in telecommunications, necessitating effective churn prediction (CP) methodologies. This paper introduces the Enhanced Gradient Boosting Model (EGBM), which uses a Support Vector Machine with a Radial Basis Function kernel (SVMRBF) as a base learner and exponential loss function to enhance the learning process of the GBM. The novel base learner significantly improves the initial classification performance of the traditional GBM and achieves enhanced performance in CP-EGBM after multiple boosting stages by utilizing state-of-the-art decision tree learners. Further, a modified version of Particle Swarm Optimization (PSO) using the consumption operator of the Artificial Ecosystem Optimization (AEO) method to prevent premature convergence of the PSO in the local optima is developed to tune the hyper-parameters of the CP-EGBM effectively. Seven open-source CP datasets are used to evaluate the performance of the developed CP-EGBM model using several quantitative evaluation metrics. The results showed that the CP-EGBM is significantly better than GBM and SVM models. Results are statistically validated using the Friedman ranking test. The proposed CP-EGBM is also compared with recently reported models in the literature. Comparative analysis with state-of-the-art models showcases CP-EGBM's promising improvements, making it a robust and effective solution for churn prediction in the telecommunications industry.


Lymphaticoperitoneal fistula.
Two cases of lymphaticoperitoneal fistula are reported. One patient had a recurrent carcinoma of the cervix and the other had a recurrent vaginal carcinoma. A brief discussion of the pathophysiology is presented.


Growth hormone promotes lymphangiogenesis.
The lymphatic system plays an important role in inflammation and cancer progression, although the molecular mechanisms involved are poorly understood. As determined using comparative transcriptional profiling studies of cultured lymphatic endothelial cells versus blood vascular endothelial cells, growth hormone receptor was expressed at much higher levels in lymphatic endothelial cells than in blood vascular endothelial cells. These findings were confirmed by quantitative real-time reverse transcriptase-polymerase chain reaction and Western blot analyses. Growth hormone induced in vitro proliferation, sprouting, tube formation, and migration of lymphatic endothelial cells, and the mitogenic effect was independent of vascular endothelial growth factor receptor-2 or -3 activation. Growth hormone also inhibited serum starvation-induced lymphatic endothelial cell apoptosis. No major alterations of lymphatic vessels were detected in the normal skin of bovine growth hormone-transgenic mice. However, transgenic delivery of growth hormone accelerated lymphatic vessel ingrowth into the granulation tissue of full-thickness skin wounds, and intradermal delivery of growth hormone resulted in enlargement and enhanced proliferation of cutaneous lymphatic vessels in wild-type mice. These results identify growth hormone as a novel lymphangiogenic factor.


Patterned dystrophies of the retinal pigment epithelium. A review.
Pigmentations and depigmentations, accompanied or not by yellow subretinal lipofuscin accumulations, are the ophthalmologically visible manifestations of dystrophies of the retinal pigment epithelium. The pigmentations may or may not become confluent and form concentric, butterfly-shaped or reticular configurations. Different patterns of pigment migration may occur in a family and even in one individual. Hence these dystrophies are called pattern(ed) dystrophies of the retinal pigment epithelium. The visual functions are relatively well-preserved, although severe visual impairment is not excluded. The pattern(ed) dystrophies are inherited as an autosomal dominant, an autosomal recessive or an X-linked recessive trait. The author suggests that some cases diagnosed as atypical pigmentary dystrophies with near-normal visual functions probably are dystrophies of the retinal pigment epithelium.


Myogenic cell movement in the developing avian limb bud in presence and absence of the apical ectodermal ridge (AER).
Fragments of quail wing bud containing myogenic cells of somitic origin and fragments of quail sphlanchopleural tissue were introduced into the interior of the wing bud of fowl embryo hosts. No movement of graft into host tissue occurred in the control, but myogenic cells from the quail wing bud fragments underwent long migrations in an apical direction to become incorporated in the developing musculature of the host. When the apical ectodermal ridge (AER), together with some subridge mesenchyme, was removed at the time of grafting, no such cell migration occurred. The capacity of grafted myogenic cells to migrate in the presence of AER persists to H.H. stage 25, when myogenesis has begun, but premyogenic cells in the somites, which normally migrate out into the early limb bud, do not migrate when somite fragments are grafted into the wing bud. Coelomic grafts of apical and proximal wing fragments showed that apical sections of quail wing buds become invaded by myogenic cells of the host, but grafts from proximal wing bud regions do not.


[Recent Advance of Newly Therapy for Chronic Myeloid Leukemia with BCR-ABLT315I Mutation--Review].
BCR-ABLT315I mutation is the main mechanism of resistance to the first and second generation tyrosine kinase inhibitor (TKI) for patients with chronic myeloid leukemia (CML). Ponatinib as the third generation TKI has been found that can significantly improve the prognosis of CML patients with T315I mutation. However, the latest report has discovered that the T315I compound mutant is even resistant to ponatinib, which aroused the enthusiasm of research on the mechanism of CML resistance and targeted therapy once again. Previous studies have shown that TKI combined with other targeted drugs is effective to CML patients with drug resistance or relapse due to T315I mutation. The latest research has found that the allosteric inhibitor asciminib combined with TKI therapy is equally effective to CML patients with T315I compound mutant, but the specific mechanism is not yet clarified. This review will focus on the latest research progress of therapy for CML with BCR-ABLT315I mutation, hoping to provide reference for researching new drugs and improve therapy for treating CML with T315I mutation.


Effects of transversus abdominis plane block versus quadratus lumborum block on postoperative analgesia: a meta-analysis of randomized controlled trials.
Trunk block technique has been used in postoperative analgesia for patients undergoing surgery, specifically, transversus abdominis plane block (TAPB) and quadratus lumborum block (QLB) have been proved effective. The purpose of this meta-analysis is to evaluate the effects of TAPB and QLB in postoperative analgesia. Online databases, including MEDLINE, EMBASE, Cochrane Library (&Trail), Web of Science, CNKI, Wanfang and QVIP were applied to collect the randomized controlled trials (RCTs) from inception to Dec. 9th, 2019. Twenty-two studies were finally included containing 777 patients in the TAPB group and 783 cases in QLB group. RCTs comparing TAPB and QLB in postoperative analgesia were included in this meta-analysis. The indicators including total analgesia consumption postoperatively, operative time, duration of anesthesia, visual analogue scale (VAS) score at 24 h postoperatively, duration of postoperative analgesia, the number of patients requiring analgesia postoperatively and adverse reactions were analyzed. our findings showed that morphine consumption (mg) (WMD = 3.893, 95%CI: 2.053 to 5.733, P < 0.001), fentanyl consumption (μg) (WMD = 23.815, 95%CI: 15.521 to 32.109, P < 0.001), VAS score at 24 h postoperatively (WMD = 0.459, 95%CI: 0.118 to 0.801, P = 0.008), the number of patients requiring analgesia postoperatively (WMD = 3.893, 95%CI: 2.053 to 5.733, P < 0.001), and the incidence of dizziness (WMD = 2.691, 95%CI: 1.653 to 4.382, P < 0.001) in TAPB group were higher than in QLB group. QLB is superior to TAPB in reducing morphine consumption, fentanyl consumption, VAS score at 24 h postoperatively, the number of patients requiring analgesia postoperatively, and the incidence of dizziness.


Emergence of Invasive Haemophilus influenzae Type A Disease in Italy.
We report on the first detection of 2 cases of invasive Haemophilus influenzae type a (Hia) disease in Italy. The cases were sustained by the same Hia "strain" belonging to the ST23 clone that has previously been reported only outside Europe. The emergence of invasive Hia disease is of concern.


Influence of epithelium and isoprenaline incubation on responsiveness of guinea-pig trachea to methacholine.
There are reports regarding harmful effect of long-term use of beta2-agonist drugs on asthma severity and airway responsiveness. In the present study, the responses of guinea pig trachea with intact and denuded epithelium (groups 1 and 2, n = 10) to methacholine as EC50 were measured in tissues nonincubated or incubated with 10 mumol/l isoprenaline during the resting period. The same protocol was performed in groups 3 and 4 (n = 5 for each group) with an additional 30 min rest time after isoprenaline incubation. The response of trachea with denuded epithelium (groups 2 and 4) to methacholine was significantly higher than that with intact epithelium both in incubated and nonincubated conditions (groups 1 and 3, p < 0.05 to p < 0.001). Incubation with isoprenaline caused a significant reduction in the tracheal response to methacholine in both the denuded groups (p < 0.005 and p < 0.001) and intact epithelium groups (p < 0.005 for both cases). The reduction in tracheal responsiveness to methacholine due to incubation in epithelium denuded trachea (groups 2 and 4) was nonsignificantly greater than that of intact epithelium tissues. There was no difference between groups 3 and 4 with those of groups 1 and 2 in both incubated and non incubated conditions. The maximum contractility response to methacholine was not different between tracheal chains with denuded and intact epithelium and did not change due to incubation with isoprenaline. The results of this study indicate reduction of tracheal response to methacholine due to incubation of tissues with isoprenaline, which was relatively more pronounced in epithelium denuded trachea.


Ulnar Nerve Management in Distal Humerus Fracture Surgery: A Case of Developing Ulnar Neuropathy After Open Reduction and Internal Fixation.
In this case report, we want to show how a patient who underwent surgery for a distal humerus fracture developed postoperative ulnar neuropathy symptoms, how nonunion persisted even at the ninth month of follow-up, and whether the nonunion was connected to the ulnar neuropathy that developed. Due to this, we used this case to explore ulnar nerve care and whether ulnar nerve transposition, manipulation, or decompression should be carried out during surgery on patients with distal humerus fractures. A 52-year-old man with a bi-columnar distal humerus fracture from a fall on his right elbow underwent open reduction and internal fixation at an external center one year before. Elbow restriction, discomfort, numbness, and weakness in the fourth and fifth digits of the right hand were all symptoms the patient experienced eight months following the surgery. We discovered the distal right humerus' nonunion during the radiological exams. It became apparent that the patient had no signs of ulnar neuropathy before the injury. In the eighth month following the injury, the patient had implant removal, open reduction internal fixation with autograft, and ulnar nerve transposition. We discovered during follow-up that the patient's ulnar neuropathy symptoms had subsided. The surgeon's familiarity with the procedure and command of the anatomy of the elbow has a role in managing the ulnar nerve in distal humerus fractures. We concluded that more study is required to determine the connection between the onset of ulnar neuropathy and nonunion while treating distal humerus fractures.


Free radicals and inflammation: superoxide-dependent activation of a neutrophil chemotactic factor in plasma.
The intravenous administration of superoxide dismutase (superoxide:superoxide oxidoreductase, EC 126.96.36.199) to animals with induced inflammation suppresses the inflammatory response and inhibits leukocyte infiltration into the challenged site, suggesting that neutrophil-generated superoxide reacts with an extracellular precursor to generate a substance chemotactic for neutrophils. Plasma exposed to superoxide in vitro becomes potently chemotactic. The appearance of chemotactic activity is inhibited by superoxide dismutase but not by catalase. The chemotactic factor does not stimulate superoxide production or degranulation in neurtrophils. Intradermal injection of superoxide-treated plasma or of a superoxide-generating system causes heavy infiltration of neutrophils to the injection site but does not cause overt signs of inflammation. The chemotactic factor consists of a chloroform-extractable component bound to serum albumin. The superoxide-dependent chemotactic factor appears to play a major role in communication in neutrophil-mediated inflammatory events. Prevention of production of this factor appears to be the major anti-inflammatory action of superoxide dismutase.


Interferon effect on collagen and fibronectin distribution in the extracellular matrix of murine sarcoma virus-transformed cells.
The transformation of murine BALB/c embryonic fibroblasts by murine sarcoma virus, Moloney strain, followed by prolonged treatment with murine interferon, resulted in the appearance of a new cell population (MSV-IF+). These MSV-IF+ cells are characterized by the recovery of a normal phenotype, contact inhibition, and lack of colony formation in agar. This phenotypic change of the MSV-IF+ cells is associated to the neosynthesis of a dense fibrous matrix beyond the cell periphery. Ultrastructural studies using peroxidase-labeled antibodies enabled us to localize the extracellular distribution of fibronectin and collagen in the MSV-IF+ cell line, compared to normal BALB/c and murine sarcoma virus-transformed cells. In parallel, a significant increase of collagen and fibronectin deposit in the intercellular space of interferon-treated murine sarcoma virus-transformed cells was observed.


Cultural similarities and differences in perceiving and recognizing facial expressions of basic emotions.
The ability to recognize facial expressions of basic emotions is often considered a universal human ability. However, recent studies have suggested that this commonality has been overestimated and that people from different cultures use different facial signals to represent expressions (Jack, Blais, Scheepers, Schyns, & Caldara, 2009; Jack, Caldara, & Schyns, 2012). We investigated this possibility by examining similarities and differences in the perception and categorization of facial expressions between Chinese and white British participants using whole-face and partial-face images. Our results showed no cultural difference in the patterns of perceptual similarity of expressions from whole-face images. When categorizing the same expressions, however, both British and Chinese participants were slightly more accurate with whole-face images of their own ethnic group. To further investigate potential strategy differences, we repeated the perceptual similarity and categorization tasks with presentation of only the upper or lower half of each face. Again, the perceptual similarity of facial expressions was similar between Chinese and British participants for both the upper and lower face regions. However, participants were slightly better at categorizing facial expressions of their own ethnic group for the lower face regions, indicating that the way in which culture shapes the categorization of facial expressions is largely driven by differences in information decoding from this part of the face.


Characterization and content of VIP and VIP mRNA in rat fore-brain neurones.
HPLC, gel chromatography, Northern analysis and non-isotopic in situ hybridization have been used to characterize for the first time the nature of vasoactive intestinal polypeptide mRNA and peptide in the rat thalamus and suprachiasmatic nucleus. Both these hitherto unstudied areas were shown to contain VIP-like immunoreactivity and VIP mRNA indistinguishable from those found in the rat cerebral cortex. Non-isotopic in situ hybridization with an alkaline phosphatase labelled antisense oligonucleotide specific for VIP mRNA and densitometry revealed that relative levels of VIP mRNA per cell were highest in the suprachiasmatic nucleus and lowest per cell in the thalamus of the rat brain areas investigated.


Strategies for adjustment of the aged.
To measure the "Strategies for Adjustment of the Aged" among the three age-group subjects, some truly effective measures are required to postpone and treat human aging in urban society by tackling it as an engineering problem. This paper describes an objective test consisting of 34 items; each item contains a statement representing a problematic situation relating to the home, health, emotional life, or social life of the subject along with four alternative responses. The responses represent four alternative strategies of adjustment; i.e., Intrapersistive Approach Mode (IAM), Extrapersistive Approach Mode (EAM), Punitive Avoidance Mode (PAM), and Defensive Avoidance Mode (DAM). The IAM was preferred among all age groups more often than any other mode to solve various problems.


[Concurrent chemoradiotherapy with sodium glycididazole and cisplatin for local advanced nasopharyngeal carcinoma].
To evaluate the radiosensitivity and toxicity of sodium glycididazole and cisplatin in concurrent chemoradiotherapy for local advanced nasopharyngeal carcinoma (NPC). Sixty patients with local advanced NPC (T3-4N2-3M0) were randomly divided into chemoradiotherapy group (n=30) and chemoradiotherapy plus sodium glycididazole group (n=30). All the patients received radiotherapy with (60)Co or 6-8 MV linear accelerator and concurrent injection of cisplatin at a weekly dose of 20 mg/m square. In sodium glycididazole group, the patients received injections of sodium glycididazole at 800 mg/m square prior to the radiotherapy 3 times a week. At the end of the therapy and 3 month after the radiotherapy, a response rate of 100% was achieved in both of the groups. But at the end of the therapy, the chemoradiotherapy plus sodium glycididazole group showed a significantly higher rate of complete tumor remission than the chemoradiotherapy group (93.3% vs 73.33%, chi(2)=4.32, P=0.038). The patients in the two groups showed similar tolerance of the therapy during the observation. Sodium glycididazole plus cisplatin can accelerate the tumor remission and improve the complete remission rate in patients with local advanced NPC without causing severe toxicity.


Experiences and impact of mistreatment and obstetric violence on women during childbearing: a systematic review protocol.
The aim of this review is to synthesize the best available evidence on the experiences of mistreatment and/or obstetric violence in women. Specifically, the objective is to explore, from a woman's point of view, the impacts and consequences of mistreatment and/or obstetric violence during childbearing. The review question is: "What are the experiences and impact of mistreatment and obstetric violence on women during the active period of childbearing?"


Perceived Concerns and Psychological Distress of Healthcare Workers Facing Three Early Stages of COVID-19 Pandemic.
This study analyzed the difference in psychological distress of the healthcare workers in three different periods of the coronavirus disease 2019 (COVID-19) pandemic in Argentina. Specifically, from the third week of the mandatory quarantine through the two following weeks. Analysis of the responses of 1,458 members of the health personnel was done on a questionnaire on healthcare workers concerns regarding the care of patients with coronavirus, indicators of depression, anxiety, intolerance of uncertainty, and coping. The psychological indicators that were considered presented differences between the evaluated periods. Perceived concerns about the possibility of infecting loved ones and infecting themselves were greatest in the periods after the onset of the pandemic. In addition, the perception of how the work environment worsened and how lack of sleep interfered with their work was also higher in periods 2 and 3. The same results were found in the indicators of depression, anxiety, and intolerance of uncertainty. Finally, the indicators of high tension and concurrent lack of emotional control, which was greater in the last periods evaluated, were also expressed in the coping strategies (showing emotional lability, only contained by hypercontrol). The differences found in the psychological indicators between the evaluated periods support the need for early psychological care of health personnel which should be a priority of public health and a fundamental fact to increase its immediate effectiveness in the care of infected patients.


Scleredema-associated IgA myeloma with myelofibrosis in a young adult: a case report.
Scleredema of Buschke is a rare fibromucinous connective tissue disorder of unknown etiology. It is often associated with a benign monoclonal gammopathy and rarely with myelomatosis. We report a case of scleredema-associated IgA myeloma with myelofibrosis in a 24-year-old male patient. Scleredema generally affects young adults and onset of associated monoclonal gammopathy is at a younger age than when not associated with scleredema. However, presentation at a much younger age (24 years in our case) is very unusual. Although mucin deposition in the bone marrow has been reported in scleredema, to the best of our knowledge, myelofibrosis has not been reported.


The illusion of a future: a friendly disagreement with Prof. Sigmund Freud.
Freud's The Future of an Illusion was not just an abstract statement of his position on religion, but part of an ongoing exchange of views with Oskar Pfister, a Zurich pastor. Freud was continuing to try to settle differences between himself and Jung. Pfister, while practising as an analyst in 1928, wrote a respectful reply to Freud, and his 'The Illusion of a Future' has never before appeared in English. Pfister was expressing what he saw as the central weaknesses in Freud's attitude toward ethics, art, philosophy, and the practice of psychotherapy.


How prepared are medical students to diagnose and manage common ocular conditions.
It is essential that primary care physicians have a solid fund of knowledge of the diagnosis and management of common eye conditions as well as ocular emergencies, as management of these diseases commonly involves appropriate referral to an ophthalmologist. Thus, it is crucial to receive comprehensive clinical knowledge of ophthalmic disease in the primary care setting during medical school. This study investigated how well prepared medical students are to diagnose and manage common ocular conditions. The study used scores from a standardized 12-question quiz administered to fourth-year medical students (N = 97; 88% response rate) and second-year medical students (N = 97; 97% response rate). The quiz comprising diagnosis and referral management questions covered the most frequently tested ophthalmology topics on board exams and assessed students' ability to recognize when referral to an ophthalmologist is appropriate. Fourth-year medical students had quiz scores ranging from 0%-94.5% with an average score of 68.7%. Second-year students had quiz scores ranging from 27.2%-86.4%, with an average score of 63.8%. Passing rate was 70%. Student's t-test showed fourth-year students had a significantly higher quiz average (P = 0.003). In general, both classes performed better on diagnostic questions (fourth-year, 73.7%; second year, 65.8%) rather than on management questions (fourth-year, 64.8%; second year, 61.8%). Both second-year and fourth-year students on average fell short on passing the ophthalmology proficiency quiz, and in general students were more adept at diagnosing rather than managing ocular conditions and emergencies.


Preferential concentration versus clustering in inertial particle transport by random velocity fields.
The concept of preferential concentration in the transport of inertial particles by random velocity fields is extended to account for the possibility of zero correlation time and compressibility of the velocity field. It is shown that, in the case of an uncorrelated in time random velocity field, preferential concentration takes the form of a condition on the field history leading to the current particle positions. This generalized form of preferential concentration appears to be a necessary condition for clustering in the uncorrelated in time case. The standard interpretation of preferential concentration is recovered considering local time averages of the velocity field. In the compressible case, preferential concentration occurs in regions of negative divergence of the field. In the incompressible case, it occurs in regions of simultaneously high strain and negative field skewness.


Separation of xiphi-omphalo-ischiopagus tetrapus twins with favorable internal anatomy.
Conjoined twinning is a rare occurrence, and ischiopagus variety is even more rare. So far, only 20 cases have been reported. The incidence is 1 in 500,000. Because of its rarity and variable anatomy, no definite line of treatment can be adapted. Time of operation and mode of treatment have to be tailored according to the local circumstances and organ sharing in each individual pair. The twins in this report were xi phi-omphalo-ischiopagus tetrapus conjoined twins. Although several internal organs were fused and shared, they were complete mirror image sets for each twin. In this case report, details of anatomy and more importantly, preoperative investigations, preparations, and organizational aspects that were designed to suit local conditions are described. Operative details, postoperative care, and short and long-term progress also has been highlighted.


Do milk-borne cytokines and hormones influence neonatal immune cell function?
Cytokines, growth factors and various hormones collectively control the proliferation, survival, differentiation and function of immune cells. A wide array of these compounds is present in maternal milk and ingested by neonates during a period of rapid maturation of gut-associated and peripheral lymphoid tissues. The functional consequences of most milk immunomodulatory constituents in neonates are unknown. However, there is evidence that milk prolactin acts as a developmental regulator of the neonatal immune system, supporting the premise that milk constituents with immunomodulatory activity may serve as neonatal immunodevelopment agents.


Carbohydrates facilitate correct disulfide bond formation and folding of rotavirus VP7.
It is well established that glycosylation is essential for assembly of enveloped viruses, but no information is yet available as to the function of carbohydrates on the nonenveloped but glycosylated rotavirus. We show that tunicamycin and, more pronouncedly, a combination of tunicamycin and brefeldin A treatment caused misfolding of the luminal VP7 protein, leading to interdisulfide bond aggregation. While formation of VP7 aggregates could be prevented under reducing conditions, they reoccurred in less than 30 min after a shift to an oxidizing milieu. Furthermore, while glycosylated VP7 interacted during maturation with protein disulfide isomerase, nonglycosylated VP7 did not, suggesting that glycosylation is a prerequisite for protein disulfide isomerase interaction. While native NSP4, which does not possess S-S bonds, was not dependent on N-linked glycosylation or on protein disulfide isomerase assistance for maturation, nonglycosylated NSP4 was surprisingly found to interact with protein disulfide isomerase, further suggesting that protein disulfide isomerase can act both as an enzyme and as a chaperone. In conclusion, our data suggest that the major function of carbohydrates on VP7 is to facilitate correct disulfide bond formation and protein folding.


Curriculum influence on interdisciplinary oral health education and practice.
Oral diseases are very prevalent across the lifespan and impact overall health, yet are largely preventable. The Smiles for Life (SFL) curriculum was created to educate healthcare providers about oral disease and support integration of oral health and primary care. This study examines SFL's influence on clinical practice and education. Surveys were sent to registered users of SFL. Users who self-identified as direct care providers (DCPs), or educators, were included in the analysis. Survey response rate was 18 percent, with 87 percent identifying as DCPs and 13 percent as educators. Across professions, 85 percent of DCPs reported SFL influencing their practice to some degree, with variance among profession type and experience. DCPs most commonly reported that SFL led them to improve how they conduct oral health activities, with 60 percent performing the activity more skillfully following completion of SFL. Fluoride varnish application was the most common practice behavior initiated, and caries risk assessments was the oral health activity affected to the greatest degree. A majority of educators (94 percent) reported that SFL led them to incorporate or enhance oral health in their teaching. SFL helped educators emphasize the importance of oral health, improved their ability to teach content, raised motivation, and reduced barriers to teaching oral health. Data supports that SFL is positively influencing oral health practice across professions, especially in areas of caries risk assessment and fluoride varnish application. SFL improves the frequency and quality with which DCPs and educators participate in oral health activities, and facilitates oral health inclusion in primary care.


The role of neuropeptide Y in the expression and extinction of fear-potentiated startle.
Neuropeptides are a promising target for novel treatments for anxiety and other psychiatric disorders and neuropeptide Y (NPY) has emerged as a key component of anxiolytic circuits in the brain. For this reason, we have evaluated the role of NPY in the expression and extinction of conditioned fear. We found that intracerebroventricular administration of NPY inhibits both baseline acoustic startle and the expression of fear-potentiated startle. Infusion of NPY (10 pmol/side) into the basolateral, but not the medial, nucleus of the amygdala reproduced the intracerebroventricular effect. Central administration of NPY (10 microg) also enhanced within-session extinction of fear-potentiated startle. This finding, coupled with the growing body of literature correlating NPY with resilience in humans, led us to the hypothesis that NPY may enhance the extinction of conditioned fear. When NPY (10 microg) is administered intracerebroventricularly before extinction training, extinction retention for both the contextual and cued components of conditioned fear is enhanced when tested 48 h later off drug. Additionally, we found that intra-basolateral amygdala administration of the NPY Y(1) receptor antagonist BIBO 3304 (200 pmol/side) before extinction training led to a profound deficit in extinction retention. This is the first evidence that NPY facilitates and an NPY antagonist blocks the extinction of conditioned fear. We believe that the role of NPY in the extinction of conditioned fear may, at least in part, explain the mechanism underlying the association between NPY and psychobiological resilience in humans.


Ectopic eyes outside the head in Xenopus tadpoles provide sensory data for light-mediated learning.
A major roadblock in the biomedical treatment of human sensory disorders, including blindness, has been an incomplete understanding of the nervous system and its ability to adapt to changes in sensory modality. Likewise, fundamental insight into the evolvability of complex functional anatomies requires understanding brain plasticity and the interaction between the nervous system and body architecture. While advances have been made in the generation of artificial and biological replacement components, the brain's ability to interpret sensory information arising from ectopic locations is not well understood. We report the use of eye primordia grafts to create ectopic eyes along the body axis of Xenopus tadpoles. These eyes are morphologically identical to native eyes and can be induced at caudal locations. Cell labeling studies reveal that eyes created in the tail send projections to the stomach and trunk. To assess function we performed light-mediated learning assays using an automated machine vision and environmental control system. The results demonstrate that ectopic eyes in the tail of Xenopus tadpoles could confer vision to the host. Thus ectopic visual organs were functional even when present at posterior locations. These data and protocols demonstrate the ability of vertebrate brains to interpret sensory input from ectopic structures and incorporate them into adaptive behavioral programs. This tractable new model for understanding the robust plasticity of the central nervous system has significant implications for regenerative medicine and sensory augmentation technology.


ExEm-FRET two-hybrid assay: FRET two-hybrid assay based on linear unmixing of excitation-emission spectra.
Simultaneous linear unmixing of excitation-emission spectra (ExEm-unmixing)-based fluorescence resonance energy transfer (FRET) two-hybrid assay method, named as ExEm-FRET two-hybrid assay, was developed for evaluating the stoichiometric ratio of macromolecular complexes in living cells. Linear unmixing of the excitation-emission spectra (SDA) of cells obtains the weight factors of donor (WD), acceptor (WA) and acceptor sensitization (WS), yielding ED and EA (donor- and acceptor-centric FRET efficiency) images. ExEm-FRET two-hybrid assay employs pixel-to-pixel titration curves of ED/EA versus the free acceptor (Ca)/donor (Cd) concentration deduced from the three weight factors to obtain EA,max and ED,max (the maximal EA and ED), thus yielding the stoichiometric ratio (EA,max/ED,max) of donor-tagged protein to acceptor-tagged protein.


Monitoring and tracking the spread of SARS-CoV-2 in Asturias, Spain.
Mutational analysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can quantify the relative importance of variants over time, enable dominant mutations to be identified, and facilitate near real-time detection, comparison and tracking of evolving variants. SARS-CoV-2 in Asturias, an autonomous community of Spain with a large ageing population, and high levels of migration and tourism, was monitored and tracked from the beginning of the pandemic in February 2020 until its decline and stabilization in August 2021, and samples were characterized using whole genomic sequencing and single nucleotide polymorphisms. Data held in the GISAID database were analysed to establish patterns in the appearance and persistence of SARS-CoV-2 strains. Only 138 non-synonymous mutations occurring in more than 1 % of the population with SARS-CoV-2 were found, identifying ten major variants worldwide (seven arose before January 2021), 19 regional and one local. In Asturias only 17 different variants were found. After vaccination, no further regional major variants were found. Only half of the defined variants circulated and no new variants were generated, indicating that infection control measures such as rapid diagnosis, isolation and vaccination were efficient.


Retrieval practice reduces relative forgetting over time.
There is overwhelming evidence in the literature that retrieval practice of studied material can lead to better final recall than restudy of the same material. Far less clear is whether this recall benefit is accompanied by reduced subsequent forgetting over time. This study revisited the issue in two experiments by comparing the effects of retrieval practice - with and without feedback -, restudy, and a no-practice condition on recall across different delay intervals ranging between three minutes and several days. We fitted power functions of time to the recall rates of each practice condition and compared relative forgetting rates between conditions. The comparisons showed that relative forgetting was reduced after retrieval practice relative to restudy, the relative forgetting rate after retrieval practice was unaffected by the presence of feedback, and forgetting after restudy did not differ from the no-practice condition. Together with other findings in the literature, the results provide evidence that retrieval practice reduces relative forgetting over time.


Comparison of estrogen receptors, progesterone receptors and HER-2/neu expression between primary and metastatic breast carcinoma.
To determine whether receptor status and HER-2/neu status remains same in primary and metastatic breast carcinoma to corresponding lymph nodes of individual patients. Estrogen receptors, progesterone receptors and HER-2/neu immunohistochemical stains were performed on primary and metastatic breast carcinoma to axillary lymph nodes in 100 patients. Data was collected on a Performa and age, tumour size, type, grade and expression of ER, PR and HER-2/neu on primary and metastatic tumours were recorded. Hormone receptor status was compared between primary and metastatic tumours. Estrogen receptor positivity was observed in 28% primary tumours which were reduced to 25% in metastatic carcinoma. Progesterone receptors were positive in 28% primary tumours as compared to 22% in metastatic tumours. Her-2/neu protein over expression was noted in 44% primary and 45% metastatic breast tumours respectively. In case to case comparison ER, PR and HER-2/neu showed 91%, 88% and 95% concordance in primary and metastatic tumours respectively. ER, PR and HER-2/neu biomarkers showed significant concordance between primary and metastatic breast carcinoma.


Effect of fluorometholone (FML) on the intraocular pressure of corticosteroid responders.
Steroid responsiveness to dexamethasone was determined in a large family with an established dominant form of chronic simple glaucoma. The high and intermediate steroid responders were then tested with fluorometholone--a steroid which is claimed to have a reduced risk of increasing intraocular pressure. It was found that fluorometholone had a much less pronounced ocular hypertensive effect than dexamethasone. However, one patient showed a comparable response on both drugs, and this suggests that some caution must still be exercised when using the drug on susceptible individuals. Fluorometholone was also found to be more irritable to the eye than dexamethasone.


Complete genomic sequence of a duck enteritis virus attenuated via serial passage in chick embryos.
Here, we present the complete genomic sequence of an attenuated duck enteritis virus (DEV). The Chinese standard challenge strain of DEV (DEV CSC) was serially passaged 20 times in chick embryo fibroblasts and then 85 times in chick embryos. The virus was attenuated and was avirulent to 2-month-old ducks. The attenuated DEV genome is 162,131 base pairs (bp) in length and as long as the parental genomic sequence. There are only 22 nucleotide substitutions, resulting in single amino acid changes in open reading frames LORF5, LORF4, UL41, UL39, UL32, UL13, UL10, UL3, US3, US4 and US7. The genome sequence has been deposited in the GenBank database under accession number KU216226. This study provides genetic information about DEV attenuation and further advances our understanding of the molecular basis of DEV pathogenesis.


Identifying Therapies to Combat Epithelial Mesenchymal Plasticity-Associated Chemoresistance to Conventional Breast Cancer Therapies Using An shRNA Library Screen.
Breast cancer (BC) is a heterogeneous disease for which the commonly used chemotherapeutic agents primarily include the anthracyclines (doxorubicin, epirubicin), microtubule inhibitors (paclitaxel, docetaxel, eribulin), and alkylating agents (cyclophosphamide). While these drugs can be highly effective, metastatic tumours are frequently refractory to treatment or become resistant upon tumour relapse. We undertook a cell polarity/epithelial mesenchymal plasticity (EMP)-enriched short hairpin RNA (shRNA) screen in MDA-MB-468 breast cancer cells to identify factors underpinning heterogeneous responses to three chemotherapeutic agents used clinically in breast cancer: Doxorubicin, docetaxel, and eribulin. shRNA-transduced cells were treated for 6 weeks with the EC10 of each drug, and shRNA representation assessed by deep sequencing. We first identified candidate genes with depleted shRNA, implying that their silencing could promote a response. Using the Broad Institute's Connectivity Map (CMap), we identified partner inhibitors targeting the identified gene families that may induce cell death in combination with doxorubicin, and tested them with all three drug treatments. In total, 259 shRNAs were depleted with doxorubicin treatment (at p < 0.01), 66 with docetaxel, and 25 with eribulin. Twenty-four depleted hairpins overlapped between doxorubicin and docetaxel, and shRNAs for TGFB2, RUNX1, CCDC80, and HYOU1 were depleted across all the three drug treatments. Inhibitors of MDM/TP53, TGFBR, and FGFR were identified by CMap as the top pharmaceutical perturbagens and we validated the combinatorial benefits of the TGFBR inhibitor (SB525334) and MDM inhibitor (RITA) with doxorubicin treatment, and also observed synergy between the inhibitor SB525334 and eribulin in MDA-MB-468 cells. Taken together, a cell polarity/EMP-enriched shRNA library screen identified relevant gene products that could be targeted alongside current chemotherapeutic agents for the treatment of invasive BC.


Thyroid Nodules in Severely Obese Patients: Frequency and Risk of Malignancy on Ultrasonography.
Background: We aimed to compare the thyroid ultrasonographic findings of severely obese versus nonobese individuals, and the frequency, characteristics, and risk of malignancy in detected nodules.Design: Case-control study including 67 adults with severe obesity (body mass index [BMI] ≥ 35 kg/m2) and 67 nonobese controls (BMI < 30 kg/m2). The participants underwent ultrasound evaluation of the thyroid and cervical subcutaneous tissue. The risk of malignancy in detected nodules was determined using the American Thyroid Association (ATA) 2015 and the Thyroid Imaging Reporting and Data System (TI-RADS) classifications. Fine-needle aspiration biopsy (FNAB) was performed in nodules for which the procedure was recommended according to the ATA-2015 or TI-RADS criteria, and the cytological evaluation followed the Bethesda classification.Results: The mean BMI values in the case and control groups were 47.0 ± 6.1 kg/m2 and 22.8 ± 2.7 kg/m2, respectively. There were no differences between groups regarding sex, age, total T3, and antiperoxidase (antiTPO) antibody positivity. When compared with controls, severely obese individuals showed a greater frequency of parenchymal hypoechogenicity (p = 0.042), cervical subcutaneous tissue thickness (p < 0.001), overall frequency of thyroid nodules (p = 0.038), and frequency of multiple nodules (p = 0.013). No significant differences were observed in terms of risk of nodular malignancy according to both the ATA-2015 and TI-RADS classifications in severely obese compared with nonobese individuals.Conclusions: Severely obese individuals (versus nonobese controls) presented increased parenchymal hypoechogenicity and frequency of thyroid nodules on ultrasonographic evaluation. However, no significant differences were observed in terms of risk of nodular malignancy between both groups according to the ATA-2015 and TI-RADS criteria. Thus, ultrasonographic thyroid screening of severely obese individuals is not justified.


Effects of sleep deprivation on human immune functions.
The effect of 40 h of wakefulness on a variety of immunological parameters in the peripheral blood from 10 normal male subjects was studied. Sleep deprivation led to enhanced nocturnal plasma interleukin 1-like and interleukin 2-like activities. The rise in nocturnal response of lymphocytes to pokeweed mitogen stimulation during a normal 24 h sleep-wake cycle was delayed by sleep deprivation, but the response to the phytohemagglutinin mitogen was unaffected. With resumed nocturnal sleep, there was a prolonged decline in natural killer cell activity (measured as spontaneous cytolytic activity for human tumor cells) and return of an increased response to pokeweed mitogen. The altered patterns in immune functions occurred independently of the cortisol circadian rhythm, which remained unchanged.


Two tautomeric forms of 2-amino-5,6-dimethylpyrimidin-4-one.
Derivatives of 4-hydroxypyrimidine are an important class of biomolecules. These compounds can undergo keto-enol tautomerization in solution, though a search of the Cambridge Structural Database shows a strong bias toward the 3H-keto tautomer in the solid state. Recrystallization of 2-amino-5,6-dimethyl-4-hydroxypyrimidine, C6H9N3O, from aqueous solution yielded triclinic crystals of the 1H-keto tautomer, denoted form (I). Though not apparent in the X-ray data, the IR spectrum suggests that small amounts of the 4-hydroxy tautomer are also present in the crystal. Monoclinic crystals of form (II), comprised of a 1:1 ratio of both the 1H-keto and the 3H-keto tautomers, were obtained from aqueous solutions containing uric acid. Forms (I) and (II) exhibit one-dimensional and three-dimensional hydrogen-bonding motifs, respectively.


Technical note: Droplet digital PCR precisely and accurately quantifies sex skew in bovine semen.
Quantifying the relative population of sperm cells bearing the X or Y chromosome in a sexed-semen sample has historically been limited to methods that are either low throughput and sensitive to user subjectivity (e.g., fluorescence in situ hybridization), conterminous (using the same technology to generate and confirm the sex skew), or relatively insensitive (including quantitative PCR with a change detection threshold of 2×). Customers pay a premium for sexed semen and should have access to reliable sex skew data, generated by an accurate, precise test that is orthogonal to the method used to generate sexed semen. Droplet digital PCR (ddPCR) has the capacity to provide an accurate and precise sex skew quantitation by subdividing a pool of template DNA into nanoliter-scale droplets containing either 1 or 0 copies of template DNA. Then PCR amplification is conducted in the droplets, and the number of copies of the amplicon of interest can be counted as the number of fluorescence-positive droplets based on classic quantitative PCR fluorescent reporters. We have optimized and validated a multiplexed ddPCR assay that uses this copy counting method to quantify the sex skew (ratio of X or Y chromosomes) in frozen-thawed bovine sexed semen. The assay interrogates at least 1,000 cells per sample well, quantifying X and Y chromosome copy numbers along with an autosomal gene, GAPDH, used as an internal assay control to confirm total cells counted. The ddPCR sex skew assay achieved a 0.5-percentage-point variance for %X or %Y with a broad linear detection range, from 10 to 95% X, and provided reproducible skew values across a range of 9 to 27 ng of genomic DNA input. This approach overcomes some limitations of other sex skew assays by quantifying absolute X and Y chromosome copy numbers, thus providing a rigorous, independent assessment of sex-skewed semen.


Predicting generalization in the training of irregular-word spelling: treating lexical spelling deficits in a child.
This paper presents a single case study investigating the mechanisms underlying generalization of treatment benefits to untrained words in spelling. Brunsdon, Coltheart, and Nickels (2005) observed that untreated words that improved tended to be those whose errors were closest to being correct prior to treatment. These words also tended to be high in written frequency. The present study employed the same treatment techniques as those used by Brunsdon et al. with K.M., a developmental surface dysgraphic. During a first treatment the characteristics of words whose spelling improved without specific training were identified. These characteristics were then used in a second treatment to test whether it was possible to predict generalization. The results showed that treatment generalization to untreated irregular words was best predicted by neighbourhood size and frequency. We suggest that the processes underlying treatment generalization are based on the interaction between the orthographic lexicon and the graphemic buffer. Clinical implications are discussed.


Correlation between grip strength and pulmonary function and respiratory muscle strength in stroke patients over 50 years of age.
Age-related skeletal muscle changes may impact respiratory muscle function, and low muscle mass is associated with low pulmonary function in older adults. Stroke not only causes weakness in the muscles of the upper and lower limbs, but it can also affect the respiratory system. This study aimed to investigate the relationships between grip strength and pulmonary function and respiratory muscle strength in stroke patients over 50 years of age. Fifty-one patients (68.69±10.40 years) who had been clinically diagnosed with ischemic or hemorrhagic stroke were included in this study. We measured these patients' forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), peak expiratory flow (PEF), maximal inspiratory pressure (MIP), maximal expiratory pressure (MEP), and hand grip strength. The data were analyzed using descriptive statistics and Pearson correlation analysis. Grip strength showed significant correlations with FVC (r=0.686, P=0.000), FEV1 (r=0.607, P=0.000), PEF (r=0.494, P=0.000), MIP (r=0.239, P=0.091), and MEP (r=0.348, P=0.012). This study demonstrated that grip strength is associated with pulmonary function and MEP in stroke patients over 50 years of age.


Small wild rodents rabies in Czechoslovakia.
During a period of 1969-1979 a total of 3174 small wild rodents of the genus Muridae and Microtinae (predominantly Microtus arvalis) were examined. The animals were trapped in five localities differing both in type of ecological conditions and epizootologic situation. Fox-rabies occurred here either enzootically, or epizootologically, or had been completely absent for many years in one of the localities investigated. Of the 71 isolated strains of RV 18 strains were isolated from brain, 25 from brown fat and 28 from salivary glands. Simultaneous isolation positivity in more than one tissue was sporadic. Virus isolations from deep-frozen material were mostly unsuccessful. Basic biological characteristics were determined in each isolate. The isolated strains differed in the rate of adaptation on albino mice and in intracerebral virulence. All isolates exhibited extraneural pathogenicity for common laboratory animals, dogs and foxes, showed distinct viscerotropism, stimulated formation of interferon and produced various forms of non-lethal infection. Incidence of "murine" rabies did not show any locality-specific differences and in longitudinally studied areas was enzootic. In a number of Czech districts there was observed a distinct relationship between fox-rabies incidence and the density of small game animal populations. Role of small wild rodents in the epizootology and epidemiology of rabies is discussed.


Evaluation of nutritional status in patients with kidney disease: usefulness of dietary recall.
Three-day food recall and normalized protein nitrogen appearance calculation from pre- and postdialysis plasma urea are the most commonly used techniques to assess nutritional intake, but a 7-day dietary recall is probably more accurate to approach dietary intake in clinical practice. A total of 99 hemodialyzed patients from two units were analyzed in a 7-day dietary record with a large range of age and without having any signs of malnutrition. Dietary protein intake was estimated from the recall and calculated (normalized protein catabolic rate) from urea kinetic modeling. Calorie intake and quality and repartition of nutrients were estimated from diaries. Repartition of nutrients was close to that of a reference population except for a lower glucidic contribution (glucide 47%, lipid 36%, protein 16%). Normalized protein catabolic rate and dietary protein intake were well correlated (R2 = 0.4), but a large variability existed from day to day, according to age (older patients are less variable) and day of dialysis (long or short interval). A large variation in alimentary intake exists from patient to patient and day to day. A 7-day evaluation of nutrient intake, dialysis adequacy, and nutritional parameters seems to be a good solution to guide dietetic counseling.


[The relationship between adenomatous hyperplasia of alveolar epithelium and bronchioalveolar carcinoma].
Surgical specimens of 7 cases of bronchioalveolar carcinoma complicated with pulmonary fibrosis and 4 cases of simple pulmonary fibrosis were examined histopathologically and immunohistochemically. The morphology and histogenesis of adenomatous hyperplasia (AH) of alveolar epithelium and its relation to bronchioalveolar carcinoma were investigated. The AH was classified as types I and II according to their microscopic morphologic characteristics. In this group, 6 cases of type I-AH and 5 cases of type II-AH were observed. The results of anti-SA, anti-SSEA-1 and anti-CEA monoclonal antibody examinations indicate that AH is a nonspecific hyperplastic lesion of alveolar epithelium occurred during chronic pulmonary inflammatory diseases. Both type I and II AH originated from type B alveolar epithelial cells. The latter developed on the basis of the former, but with a more immature tendency and hyperplastic potential, being a pre-malignant alteration. It could be considered that certain cases of bronchioalveolar carcinoma are originated from type B alveolar epithelial cells, some of which underwent malignant change from type II alveolar hyperplasia.


Transcription factor control of asymmetric cell divisions that establish the stomatal lineage.
The establishment of new cell lineages during development often requires a symmetry-breaking event. An asymmetric division in the epidermis of plants initiates a lineage that ultimately produces stomatal guard cells. Stomata are pores in the epidermis that serve as the main conduits for gas exchange between plants and the atmosphere; they are critical for photosynthesis and exert a major influence on global carbon and water cycles. Recent studies implicated intercellular signalling in preventing the inappropriate production of stomatal complexes. Genes required to make stomata, however, remained elusive. Here we report the identification of a gene, SPEECHLESS (SPCH), encoding a basic helix-loop-helix (bHLH) transcription factor that is necessary and sufficient for the asymmetric divisions that establish the stomatal lineage in Arabidopsis thaliana. We demonstrate that SPCH and two paralogues are successively required for the initiation, proliferation and terminal differentiation of cells in the stomatal lineage. The stomatal bHLHs define a molecular pathway sufficient to create one of the key cell types in plants. Similar molecules and regulatory mechanisms are used during muscle and neural development, highlighting a conserved use of closely related bHLHs for cell fate specification and differentiation.


Life adjustment correlates of physical self-concepts.
This research tested relationships between physical self-concepts and contemporary measures of life adjustment. University students (119 females, 126 males) completed the Physical Self-Perception Profile assessing self-concepts of sport competence, physical condition, attractive body, strength, and general physical self-worth. Multiple regression found significant associations (P < 0.05 to P < 0.001) in hypothesized directions between physical self-concepts and positive affect, negative affect, depression, and health complaints in 17 of 20 analyses. Thirteen of these relationships remained significant when controlling for the Bonferroni effect. Hierarchical multiple regression examined the unique contribution of physical self-perceptions in predicting each adjustment variable after accounting for the effects of global self-esteem and two measures of social desirability. Physical self-concepts significantly improved associations with life adjustment (P < 0.05 to P < 0.05) in three of the eight analyses across gender and approached significance in three others. These data demonstrate that self-perceptions of physical competence in college students are essentially related to life adjustment, independent of the effects of social desirability and global self-esteem. These links are mainly with perceptions of sport competence in males and with perceptions of physical condition, attractive body, and general physical self-worth in both males and females.


B7-H3-targeting Fc-optimized antibody for induction of NK cell reactivity against sarcoma.
Natural killer (NK) cells largely contribute to antibody-dependent cellular cytotoxicity (ADCC), a central factor for success of monoclonal antibodies (mAbs) treatment of cancer. The B7 family member B7-H3 (CD276) recently receives intense interest as a novel promising target antigen for immunotherapy. B7-H3 is highly expressed in many tumor entities, whereas expression on healthy tissues is rather limited. We here studied expression of B7-H3 in sarcoma, and found substantial levels to be expressed in various bone and soft-tissue sarcoma subtypes. To date, only few immunotherapeutic options for treatment of sarcomas that are limited to a minority of patients are available. We here used a B7-H3 mAb to generate chimeric mAbs containing either a wildtype Fc-part (8H8_WT) or a variant Fc part with amino-acid substitutions (S239D/I332E) to increase affinity for CD16 expressing NK cells (8H8_SDIE). In comparative studies we found that 8H8_SDIE triggers profound NK cell functions such as activation, degranulation, secretion of IFNγ and release of NK effector molecules, resulting in potent lysis of different sarcoma cells and primary sarcoma cells derived from patients. Our findings emphasize the potential of 8H8_SDIE as novel compound for treatment of sarcomas, particularly since B7-H3 is expressed in bone and soft-tissue sarcoma independent of their subtype.


Megakaryocytes respond during sepsis and display innate immune cell behaviors.
Megakaryocytes (MKs) are precursors to platelets, the second most abundant cells in the peripheral circulation. However, while platelets are known to participate in immune responses and play significant functions during infections, the role of MKs within the immune system remains largely unexplored. Histological studies of sepsis patients identified increased nucleated CD61+ cells (MKs) in the lungs, and CD61+ staining (likely platelets within microthrombi) in the kidneys, which correlated with the development of organ dysfunction. Detailed imaging cytometry of peripheral blood from patients with sepsis found significantly higher MK counts, which we predict would likely be misclassified by automated hematology analyzers as leukocytes. Utilizing in vitro techniques, we show that both stem cell derived MKs (SC MKs) and cells from the human megakaryoblastic leukemia cell line, Meg-01, undergo chemotaxis, interact with bacteria, and are capable of releasing chromatin webs in response to various pathogenic stimuli. Together, our observations suggest that MK cells display some basic innate immune cell behaviors and may actively respond and play functional roles in the pathophysiology of sepsis.


Determination by GC×GC of fatty acid and conjugated linoleic acid (CLA) isomer profiles in six selected tissues of lambs fed on pasture or on indoor diets with and without rumen-protected CLA.
In this study GC×GC was used to study the effects of pasture, hay, concentrate (indoor), and indoor plus 8 g/day of a rumen-protected conjugated linoleic acid (indoor-CLA) diets on the detailed fatty acid (FA) profiles of six tissues (muscles, fatty tissues, and liver) collected from 36 lambs. This powerful technique allowed the quantification of 128 FAs, of which 21 SFAs, 16 MUFAs, 19 PUFAs were identified by reference standards. The diets had similar, but not identical, effects on FA profiles (g/100 g FA) in the various tissues, as both indoor diets reduced total PUFAs (from 8.91 ± 6.27 to 8.06 ± 5.97; p < 0.05) and n-3 PUFAs (from 2.70 ± 2.37 to 1.50 ± 1.69; p < 0.01) and increased n-6 PUFA (from 3.76 ± 2.46 to 4.58 ± 3.42; p < 0.01), branched (from 2.37 ± 2.05 to 3.23 ± 0.54; p < 0.01), odd-chain FAs (from 5.88 ± 5.33 to 7.07 ± 1.51; p < 0.01) compared to pasture. Indoor-CLA increased CLAc9,t11 (from 0.42 ± 0.13 to 0.53 ± 0.19; p < 0.01), CLAt10,c12 (from 0.07 ± 0.06 to 0.12 ± 0.22; p < 0.05), and CLAc11,t13 (from 0.02 ± 0.04 to 0.05 ± 0.04; p < 0.05) compared to indoor.


Sleep deprivation in the rat: VII. Immune function.
Immune function studies were performed on splenic lymphocytes obtained from rats subjected to total or paradoxical sleep deprivation. Spleen cell counts, in vitro lymphocyte proliferation responses to mitogens, and in vitro and in vivo plaque-forming cell responses to antigens were obtained. Sleep-deprived rats were roughly equivalent to both their yoked controls and home-cage controls in all assays. The results do not support the hypothesis that sleep deprivation results in immune suppression as measured by the above-mentioned parameters.


Human semi-supervised learning.
Most empirical work in human categorization has studied learning in either fully supervised or fully unsupervised scenarios. Most real-world learning scenarios, however, are semi-supervised: Learners receive a great deal of unlabeled information from the world, coupled with occasional experiences in which items are directly labeled by a knowledgeable source. A large body of work in machine learning has investigated how learning can exploit both labeled and unlabeled data provided to a learner. Using equivalences between models found in human categorization and machine learning research, we explain how these semi-supervised techniques can be applied to human learning. A series of experiments are described which show that semi-supervised learning models prove useful for explaining human behavior when exposed to both labeled and unlabeled data. We then discuss some machine learning models that do not have familiar human categorization counterparts. Finally, we discuss some challenges yet to be addressed in the use of semi-supervised models for modeling human categorization.


Role of Semaphorins in Ischemic Stroke.
Ischemic stroke is one of the major causes of neurological morbidity and mortality in the world. Although the management of ischemic stroke has been improved significantly, it still imposes a huge burden on the health and property. The integrity of the neurovascular unit (NVU) is closely related with the prognosis of ischemic stroke. Growing evidence has shown that semaphorins, a family of axon guidance cues, play a pivotal role in multiple pathophysiological processes in NVU after ischemia, such as regulating the immune system, angiogenesis, and neuroprotection. Modulating the NVU function via semaphorin signaling has a potential to develop a novel therapeutic strategy for ischemic stroke. We, therefore, review recent progresses on the role of semphorin family members in neurons, glial cells and vasculature after ischemic stroke.


Synergistic in-vitro effects of combining an antiglycolytic, 3-bromopyruvate, and a bromodomain-4 inhibitor on U937 myeloid leukemia cells.
This project investigated the in-vitro effects of a glycolytic inhibitor, 3-bromopyruvate (3-BrP), in combination with and a new in silico-designed inhibitor of the bromodomain-4 (BRD-4) protein, ITH-47, on the U937 acute myeloid leukemia cell line. 3-BrP is an agent that targets the altered metabolism of cancer cells by interfering with glucose metabolism in the glycolytic pathway. ITH-47 is an acetyl-lysine inhibitor that displaces bromdomain 4 proteins from chromatin by competitively binding to the acetyl-lysine recognition pocket of this bromodomain and extraterminal (BET) BRD protein, thereby preventing transcription of cancer-associated genes and further cell growth. Cell growth studies determined the IC50 after 48 h exposure for 3-BrP and ITH-47 to be 6 and 2 μmol/l, respectively. When combined, 2.4 and 1 μmol/l of 3-BrP and ITH-47, respectively, inhibited 50% of the cell population, yielding a synergistic combination index of 0.9. Subsequent mechanistic studies showed that the IC50 concentrations of ITH-47 and 3-BrP and the combination increased observable apoptotic bodies and cell shrinkage in U937 cells treated for 48 h. Cell cycle analysis showed an increase in the sub-G1 fraction in all treated cells, suggesting that cell death was increased in the treated samples. Annexin-V-FITC apoptosis analysis showed a statistically significant increase in the number of cells in early and late apoptosis, indicating that cell death occurred through apoptosis and not necrosis. Only U937 cells exposed to ITH-47 showed a decrease in mitochondrial membrane potential compared with the vehicle control. Reactive oxygen species production was decreased in all treated samples. ITH-47-exposed cells showed a decrease in c-Myc, Bcl-2, and p53 gene expressions. 3-BrP-treated cells showed an increase in c-myc and p53 gene expressions. The combination of ITH-47 and 3-BrP lead to downregulation of c-myc and Bcl-2 genes. ITH-47 exposure conditions yielded a marked decrease in c-myc protein levels as well as a decrease in Ser70 phosphorylated Bcl-2. Analysis of 3-BrP and the combination of ITH-47 and 3-BrP test conditions indicated an increase in p53 protein levels. This novel study is the first to investigate the in-vitro synergistic therapeutic effect of ITH-47 and 3-BrP. The current study contributes toward unraveling the in-vitro molecular mechanisms and signal transduction associated with a novel combination of BRD inhibitors and antiglycolytic agents, providing a basis for further research on these combinations.