Inhibition of Zika virus by Wolbachia in Aedes aegypti.
Through association with cases of microcephaly in 2015, Zika virus (ZIKV) has transitioned from a relatively unknown mosquito-transmitted pathogen to a global health emergency, emphasizing the need to improve existing mosquito control programs to prevent future disease outbreaks. The response to Zika must involve a paradigm shift from traditional to novel methods of mosquito control, and according to the World Health Organization should incorporate the release of mosquitoes infected with the bacterial endosymbiont Wolbachiapipientis. In our recent paper [Dutra, HLC et al., Cell Host & Microbe 2016] we investigated the potential of Wolbachia infections in Aedes aegypti to restrict infection and transmission of Zika virus recently isolated in Brazil. Wolbachia is now well known for its ability to block or reduce infection with a variety of pathogens in different mosquito species including the dengue (DENV), yellow fever, and chikungunya viruses, and malaria-causing Plasmodium, and consequently has great potential to control mosquito-transmitted diseases across the globe. Our results demonstrated that the wMel Wolbachia strain in Brazilian Ae. aegypti is a strong inhibitor of ZIKV infection, and furthermore appears to prevent transmission of infectious viral particles in mosquito saliva, which highlights the bacterium's suitability for more widespread use in Zika control.


Diabetes education self-management intervention in improving self-efficacy for people with type 2 diabetes in the Gulf Cooperation Council countries: A systematic review.
This research aims to evaluate the effectiveness of diabetes education self-management intervention for improving self-efficacy for people with type 2 diabetes mellitus in the Gulf Cooperation Council countries. The Joanna Briggs Institute's methodology was applied to conduct a systematic review of type 2 diabetes interventions published from 2012 to 2022. This involved searching major databases, including Ovid, CINAHL, Scopus, Web of Science, and PubMed, with specific inclusion and exclusion criteria. The focus was on studies involving adults aged 18 or older with a Type 2 Diabetes (T2D) diagnosis, randomized controlled trials, and full-text English-language articles, while excluding materials such as editorials and conference abstracts. A total of 689 relevant articles initially retrieved; ultimately, only five studies met the inclusion criteria of this review. All five included studies were randomised control trials, and all five studies indicated that there was a significant and positive impact of diabetes education self-management intervention on diet and physical activities. Furthermore, three of the included studies indicated that interventions led to a substantial reduction in the levels of glycosylated haemoglobin (HbA1c) among people with type 2 diabetes. Diabetes education is an effective way of improving the health outcomes of people with type 2 diabetes. Diabetes education self-management interventions significantly increase the self-efficacy of people with Type 2 Diabetes (T2D) by enabling them to effectively control their blood glucose levels. Therefore, this study recommended that the GCC countries need to implement education intervention programmes to help and support people with Type 2 Diabetes.


A rapid and sensitive screening system for human type I collagen with the aim of discovering potent anti-aging or anti-fibrotic compounds.
This study was undertaken with the aim of developing an easy and quick means of analyzing the effect of various compounds on the synthesis and secretion of human type I collagen at the protein level. A modification of the ELISA method was used on HFF-1 cells. For the proof of concept, we used thirteen compounds most of which are known to be antioxidants. Each compound was tested at concentrations of 0, 10 and 100 microM on HFF-1 cells for 24 h. Thirteen sets of experiments for each compound were performed in ANOVA with three replicates. Duncan multiple range test (DMRT) was used to compare the mean values obtained from the treatment groups. From the results it was concluded that Vitamin C, undecylenic acid, conjugated linoleic acid, glycolic acid, and citric acid at 100 microM concentration could be used for anti-wrinkling or protection from premature aging, which requires enhancement of collagen synthesis. Lactic acid, EGCG, resveratrol, and retinol that can inhibit collagen synthesis effectively in a dose-dependent manner may be used for anti-fibrosis treatment purposes.


Reform the watchword as OECD countries struggle to contain health care costs.
Like other countries in the Organization for Economic Cooperation and Development, Canada is attempting to contain the overall costs of health care by concentrating on the reform of health care delivery, not health care financing. Systems like Canada's, with predominantly public financing and mainly private delivery, have become increasingly popular around the world. Like other nations, Canada has implemented reforms to make the most of the health care dollars we do have. In this article, Susan MacPhee examines common approaches to health care reform.


Construction of a simple suction electrode for extracellular recording and stimulation.
Principles of signal transmission in nervous systems are commonly demonstrated in the undergraduate neuroscience laboratory through extracellular recording of nerve and muscle action potentials. Here we describe the construction of a simple suction electrode that we use routinely in our laboratory classes for nerve recording and stimulation. The electrode parts are relatively inexpensive, easily available from established scientific and electronic distributors and local hardware stores, and the electrode is resilient to student handling. Our undergraduate students use this electrode design for high resolution, extracellular recordings of action potentials from crayfish motor and sensory nerves and insect muscle, and for stimulation of crustacean and insect motor nerves.


A new role for RPTPsigma in spinal cord injury: signaling chondroitin sulfate proteoglycan inhibition.
It has been known for more than two decades that chondroitin sulfate proteoglycans (CSPGs) inhibit axonal growth and regeneration. In the adult nervous system, CSPGs are enriched in perineuronal nets, and their abundance is increased in reactive astrocytes following injury to brain or spinal cord. Degradation of chondroitin sulfate (CS) sugar moieties by the local infusion of the bacterial enzyme chondroitinase ABC (ChaseABC) enhances experience-dependent neuronal plasticity in the adult visual cortex and results in substantially improved behavioral outcomes after spinal cord injury (SCI). Although the positive effects of ChaseABC treatment on neuronal plasticity have been known for some time, the underlying mechanisms remained enigmatic. The receptor protein tyrosine phosphatase sigma (RPTPsigma) has now been identified as a receptor for inhibitory CSPGs. Similarly to ChaseABC treatment, functional ablation of Ptprs, the gene encoding RPTPsigma, promotes neurite outgrowth in the presence of CSPGs in vitro and enhances axonal growth into CSPG-rich scar tissue following SCI in vivo. The discovery of neuronal RPTPsigma as a receptor for inhibitory CSPGs not only provides important mechanistic clues about CSPG function, but also identifies a potential new target for enhancing axonal growth and plasticity after nervous system injury.


Risk Stratification for Supratherapeutic Peak Anti-Xa Levels in Adult Patients on Therapeutic Enoxaparin.
The American Society of Hematology Guidelines for the management of venous thromboembolism recommend against the use of anti-Xa monitoring for assessing enoxaparin dosing based on a low level of evidence associating supratherapeutic levels with an increased risk of bleeding. However, institutions still utilize anti-Xa levels in select patient populations with altered volume of distribution and/or excretion to monitor and adjust therapy. The primary objective of this study was to identify risk factors associated with supratherapeutic peak anti-Xa levels (≥1.10 IU/mL) for patients receiving therapeutic enoxaparin. This was a retrospective single-center study performed at an academic tertiary care hospital. Patients who received enoxaparin at 1 mg/kg twice daily and peak anti-Xa monitoring were separated into supratherapeutic and therapeutic/subtherapeutic cohorts. A total of 436 patients were screened, and 215 were included, with a mean age of 62 years. There were 108 in the therapeutic/subtherapeutic cohort and 107 in the supratherapeutic cohort. Acute kidney injury (AKI), body mass index (BMI), weight, female sex, intensive care unit (ICU) service, Sequential Organ Failure Assessment (SOFA) score ≥4, and creatinine clearance at the time of peak anti-Xa level collection were associated with supratherapeutic anti-Xa levels in univariate models. Adjusted logistic regression models were created and identified BMI in the 30 to 34.9 kg/m2 (odds ratio [OR] 4.35; 95% confidence interval [CI] 1.70-11.13, P < 0.005) and ≥35 kg/m2 (OR 6.75; 95% CI 3.05-14.94, P < 0.005) and AKI (OR 2.62; 95% CI 1.04-6.62, P = 0.042) as significant risk factors for supratherapeutic anti-Xa levels. Our study identified BMI ≥ 30 kg/m2, AKI, female sex, ICU service, SOFA score ≥4, and creatinine clearance as risk factors for supratherapeutic anti-Xa levels in patients receiving 1 mg/kg twice daily dosing of enoxaparin. Further research should be done to provide evidence for the association between anti-Xa levels and bleeding risk.


Cellular underpinnings of the selective vulnerability to tauopathic insults in Alzheimer's disease.
Neurodegenerative diseases such as Alzheimer's disease (AD) exhibit pathological changes in the brain that proceed in a stereotyped and regionally specific fashion, but the cellular and molecular underpinnings of regional vulnerability are currently poorly understood. Recent work has identified certain subpopulations of neurons in a few focal regions of interest, such as the entorhinal cortex, that are selectively vulnerable to tau pathology in AD. However, the cellular underpinnings of regional susceptibility to tau pathology are currently unknown, primarily because whole-brain maps of a comprehensive collection of cell types have been inaccessible. Here, we deployed a recent cell-type mapping pipeline, Matrix Inversion and Subset Selection (MISS), to determine the brain-wide distributions of pan-hippocampal and neocortical neuronal and non-neuronal cells in the mouse using recently available single-cell RNA sequencing (scRNAseq) data. We then performed a robust set of analyses to identify general principles of cell-type-based selective vulnerability using these cell-type distributions, utilizing 5 transgenic mouse studies that quantified regional tau in 12 distinct PS19 mouse models. Using our approach, which constitutes the broadest exploration of whole-brain selective vulnerability to date, we were able to discover cell types and cell-type classes that conferred vulnerability and resilience to tau pathology. Hippocampal glutamatergic neurons as a whole were strongly positively associated with regional tau deposition, suggesting vulnerability, while cortical glutamatergic and GABAergic neurons were negatively associated. Among glia, we identified oligodendrocytes as the single-most strongly negatively associated cell type, whereas microglia were consistently positively correlated. Strikingly, we found that there was no association between the gene expression relationships between cell types and their vulnerability or resilience to tau pathology. When we looked at the explanatory power of cell types versus GWAS-identified AD risk genes, cell type distributions were consistently more predictive of end-timepoint tau pathology than regional gene expression. To understand the functional enrichment patterns of the genes that were markers of the identified vulnerable or resilient cell types, we performed gene ontology analysis. We found that the genes that are directly correlated to tau pathology are functionally distinct from those that constitutively embody the vulnerable cells. In short, we have demonstrated that regional cell-type composition is a compelling explanation for the selective vulnerability observed in tauopathic diseases at a whole-brain level and is distinct from that conferred by risk genes. These findings may have implications in identifying cell-type-based therapeutic targets.


Myocardial kinetics of carbon-11-epinephrine in the isolated working rat heart.
The kinetics of EPI were studied in the isolated rat heart model to evaluate 11C-epinephrine (EPI) as a radiotracer for the assessment of sympathetic neuronal function in the heart. Isolated rat hearts were perfused in a working mode. Carbon-11-EPI was added to the perfusate during wash-in period of 20 min, followed by a washout period of 40 min. Radioactivity in the heart was externally monitored and time-activity curves were recorded as a function of time. Effluent samples were collected throughout each study to determine the fraction of 11C radioactivity as intact tracer. Time-activity curves of control hearts showed that 11C-EPI is taken up and retained by the myocardium. Desipramine inhibition (DMI) of uptake-1 resulted in a significant decrease in myocardial uptake and retention of 11C-EPI by 91% compared to controls. Addition of DMI to the perfusion medium during washout did not affect kinetics of 11C-EPI compared to control hearts. Reserpine pretreated rat hearts also showed significant decrease in tracer retention of 95% compared to controls. The metabolic data showed that, in control conditions, about 61% of 11C-EPI taken up by the rat heart is rapidly metabolized and released. Carbon-11-EPI traces sympathetic nerve terminals in the isolated rat heart. Uptake blockade by DMI and reserpine suggest that uptake and storage of 11C-EPI appear to be similar to that of norepinephrine. However, the prominent metabolic pathway warrants further consideration. These results suggest that 11C-EPI may be a suitable radiolabeled tracer for the evaluation of sympathetic vesicular function of the heart by PET.


Visualization of thalamic nuclei on high resolution, multi-averaged T1 and T2 maps acquired at 1.5 T.
The ability to differentiate noninvasively between the primary nuclear divisions of the thalamus has immediate clinical applicability for surgical planning and guidance of functional stereotactic procedures. Comparison of prior qualitative magnetic resonance imaging (MRI) studies carried out at field strengths of 1.5 and 4 Tesla have revealed contrast within the thalamus that varies with field strength, suggesting possible differences in the inherent T1 and T2 relaxation times of the constituent nuclei. We investigate this hypothesis through acquisition of high-resolution, multi-averaged deep-brain T1 and T2 maps of a healthy volunteer. Fourteen nuclei were identified using their center-of-mass coordinates (in Talairach space) and average T1 and T2 values obtained from regions of interest placed within each. Results from this analysis revealed significant differences in T1 and T2 between the nuclei with a T1 range from 700 to 1,400 ms and a T2 range from 89 to 122 ms, allowing visual discrimination between the major nuclei groups. Furthermore, the high-resolution images showed distinct borders of T1 and T2 hypointensity surrounding each nucleus, revealing structure not reported previously. These results confirm our hypothesis and demonstrate the potential high-resolution quantitative imaging for nucleus visualization and surgical planning.


Functional analysis of the five melon necrotic spot virus genome-encoded proteins.
Function of the melon necrotic spot virus (MNSV) genome-encoded proteins (p29, p89, p7A, p7B and p42) has been studied. Protein-expression mutants of an infectious, full-length cDNA clone of a Spanish MNSV-Al isolate and a recombinant green fluorescent protein (GFP)-expressing virus were used in infection bioassays on melon plants. Results revealed that p29 and p89 are both essential for virus replication, whereas small proteins p7A and p7B are sufficient to support viral movement between adjacent cells operating in trans. It is also demonstrated that, in addition to its structural role as coat protein, p42 is an important factor controlling symptoms and is required for systemic transport. Moreover, both p42 and p7B, among all of the MNSV-encoded proteins, were able to delay RNA silencing in transient-expression assays on GFP-transgenic Nicotiana benthamiana plants. Finally, the presence of p42 also produced an enhancing effect on local spread similar to that of potyviral helper component proteinase (HC-Pro), probably due to its RNA silencing-suppression ability.


Effect of chinese herbal medicine "saiko-keishi-to" on transmembrane ionic current of snail neurons.
The effect of the Chinese herbal medicine "Saiko-Keishi-To" (SK) on transmembrane ionic current was examined to elucidate the mechanism of the anticonvulsant action of SK. SK caused a decrease in tramsmembarane sodium, calcium and potassium current. SK was also found to inhibit the negative resistance characteristics in an experiment using a voltage clamp with triangular command.


High Dose of Atorvastatin for the Treatment of Contrast-Induced Nephropathy After Carotid Artery Stenting.
Statins have been used to prevent contrast-induced nephropathy (CIN). However, the optimal dose of statins is still under controversy. This study aimed to investigate the optimal dose of atorvastatin for the treatment of CIN after carotid artery stenting (CAS). Seventy-six patients receiving selective CAS were randomized to receive 3 different dose of atorvastatin (low dose, 20 mg, n = 30; intermediate dose, 40 mg, n = 24; high dose, 60 mg, n = 22). Preoperatively and on day 3 postoperatively, the levels of serum creatinine, blood urea nitrogen, high-sensitivity C-reactive protein (hs-CRP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatine kinase (CK) were measured. Creatinine clearance (Ccr) and CIN incidence were calculated. In patients treated with high-dose atorvastatin, no significant change was observed in levels of serum creatinine (Scr), blood urea nitrogen (BUN), creatinine clearance, and high-sensitivity C-reactive protein after the CAS procedure (P > 0.05). The CIN incidence in the high-dose group (0%) was significantly lower than the low-dose (13.3%) and intermediate (8.3%) groups (P < 0.05). In the high-dose group, levels of alanine aminotransferase, aspartate aminotransferase, and creatine kinase were significantly increased after CAS (P < 0.05). Pretreatment with 40 mg of atorvastatin is both effective and safe in preventing CIN after CAS. Adverse events of the live and heart should be closely monitored during atorvastatin treatment.


Use of aspartame in pregnancy.
The low-calorie sweetening agent, aspartame, is broken down in the small intestine into three moieties: aspartic acid, methanol and phenylalanine. Acute loading studies have been performed in human beings who received up to six times the 99th percentile of the projected daily intake (6 X 34 = 200 mg/kg). No evidence of risk to the fetus was developed. Aspartate does not readily cross the placenta. Small elevations of blood methanol following such abuse doses of aspartame did not lead to measurable increases of blood formic acid, which is the product responsible for the acidosis and ocular toxicity in methanol poisoning. Phenylalanine is concentrated on the fetal side of the placenta. Aspartame in abuse doses up to 200 mg/kg in normal subjects, or to 100 mg/kg in PKU heterozygotes, did not raise blood phenylalanine levels to the range generally accepted to be associated with mental retardation in the offspring. It is concluded that, under foreseeable conditions of use, aspartame poses no risk for use in pregnancy.


Post-traumatic Recovery to Distress Symptoms Ratio Mediates Relations of Resilience Fostering Resources and Their Predictors.
A large-scale study investigated the direct and indirect effects of demographic predictors and traumatic experiences on resilience fostering resources following a war. The sample consisted of 829 Israeli adults, living in a border town, 1 year after experiencing the 2006 war with Lebanon. Resilience was assessed by measures of individual and public resilience and low sense of danger. Results show that as hypothesized the proportion of post-traumatic recovery to post-war distress symptoms predicts these indices of resilience and partly mediates the direct links between these indices and demographic predictors (age, gender and economic condition) and exposure to war. Research of resilience has proposed several prototypical trajectories that characterize responses to potentially traumatic events. Our discussion suggests that these trajectories and their changes over time can be accounted for by the balance of post-war recovery to symptoms level. Copyright © 2014 John Wiley & Sons, Ltd.


Clinical characteristics and outcome of critically ill COVID-19 patients with acute kidney injury: a single centre cohort study.
Acute kidney injury (AKI) is a common manifestation among patients critically ill with SARS-CoV-2 infection (Coronavirus 2019) and is associated with significant morbidity and mortality. The pathophysiology of renal failure in this context is not fully understood, but likely to be multifactorial. The intensive care unit outcomes of patients following COVID-19 acute critical illness with associated AKI have not been fully explored. We conducted a cohort study to investigate the risk factors for acute kidney injury in patients admitted to and intensive care unit with COVID-19, its incidence and associated outcomes. We reviewed the medical records of all patients admitted to our adult intensive care unit suffering from SARS-CoV-2 infection from 14th March 2020 until 12th May 2020. Acute kidney injury was defined using the Kidney Disease Improving Global Outcome (KDIGO) criteria. The outcome analysis was assessed up to date as 3rd of September 2020. A total of 81 patients admitted during this period. All patients had acute hypoxic respiratory failure and needed either noninvasive or invasive mechanical ventilatory support. Thirty-six patients (44%) had evidence of AKI (Stage I-33%, Stage II-22%, Renal Replacement Therapy (RRT)-44%). All patients with AKI stage III had RRT. Age, diabetes mellitus, immunosuppression, lymphopenia, high D-Dimer levels, increased APACHE II and SOFA scores, invasive mechanical ventilation and use of inotropic or vasopressor support were significantly associated with AKI. The peak AKI was at day 4 and mean duration of RRT was 12.5 days. The mortality was 25% for the AKI group compared to 6.7% in those without AKI. Among those received RRT and survived their illness, the renal function recovery is complete and back to baseline in all patients. Acute kidney injury and renal replacement therapy is common in critically ill patients presenting with COVID-19. It is associated with increased severity of illness on admission to ICU, increased mortality and prolonged ICU and hospital length of stay. Recovery of renal function was complete in all survived patients.


NFE2 Induces miR-423-5p to Promote Gluconeogenesis and Hyperglycemia by Repressing the Hepatic FAM3A-ATP-Akt Pathway.
Hepatic FAM3A expression is repressed under obese conditions, but the underlying mechanism remains unknown. This study determined the role and mechanism of miR-423-5p in hepatic glucose and lipid metabolism by repressing FAM3A expression. miR-423-5p expression was increased in the livers of obese diabetic mice and in patients with nonalcoholic fatty liver disease (NAFLD) with decreased FAM3A expression. miR-423-5p directly targeted FAM3A mRNA to repress its expression and the FAM3A-ATP-Akt pathway in cultured hepatocytes. Hepatic miR-423-5p inhibition suppressed gluconeogenesis and improved insulin resistance, hyperglycemia, and fatty liver in obese diabetic mice. In contrast, hepatic miR-423-5p overexpression promoted gluconeogenesis and hyperglycemia and increased lipid deposition in normal mice. miR-423-5p inhibition activated the FAM3A-ATP-Akt pathway and repressed gluconeogenic and lipogenic gene expression in diabetic mouse livers. The miR-423 precursor gene was further shown to be a target gene of NFE2, which induced miR-423-5p expression to repress the FAM3A-ATP-Akt pathway in cultured hepatocytes. Hepatic NFE2 overexpression upregulated miR-423-5p to repress the FAM3A-ATP-Akt pathway, promoting gluconeogenesis and lipid deposition and causing hyperglycemia in normal mice. In conclusion, under the obese condition, activation of the hepatic NFE2/miR-423-5p axis plays important roles in the progression of type 2 diabetes and NAFLD by repressing the FAM3A-ATP-Akt signaling pathway.


Methods of punishment used by Turkish mothers of preschoolers.
The goal of this study was to describe the punishments that Turkish mothers use with their preschool-aged children. 129 mothers with children between 3 and 6 years old were included in the study. Mothers completed a data collection form prepared by the researcher. Punishments mothers applied and mothers' age and education and children's age and sex were examined by chi-squared analysis. Mothers used various punishments with children. Some punishments were used with high frequency. Mothers' age and education and children's age and sex were not related to types of punishment used.


Well-differentiated neurocytoma: what is the best available treatment?
Most neurocytomas are well differentiated, being associated with better long-term survival than the more aggressive atypical lesions. Atypical neurocytomas are characterized by an MIB-1 labeling index >3% or atypical histologic features. This analysis focuses on well differentiated neurocytomas in order to define the optimal treatment. A case with a follow-up of 132 months is presented. The patient developed two recurrences two and four years after first surgery, each showing an increasing proliferation activity. Furthermore, all published well-differentiated neurocytoma cases were reviewed for surgery, radiotherapy, and prognosis. Additional relevant data were obtained from the authors. Complete resection (CTR), complete resection plus radiotherapy (CTR + RT), incomplete resection (ITR), and incomplete resection plus radiotherapy (ITR + RT) were compared for outcome by using the Kaplan-Meier method and the log-rank test. Data were complete in 301 patients (CTR, 108; CTR + RT, 27; ITR, 81; ITR + RT, 85). Local control and survival were better after CTR than after ITR (P < 0.0001 and P = 0.0085, respectively). Radiotherapy improved local control after ITR (P < 0.0001) and after CTR (P = 0.0474), but not survival (P = 0.17 and P = 1.0, respectively). In the ITR + RT group, doses < or =54 Gy (n = 33) and >54 Gy (n = 32) were not significantly different for local control (P = 0.88) and survival (P = 0.95). The data demonstrated CTR to be superior to ITR for local control and survival. After CTR and ITR, radiotherapy improved local control, but not survival. A radiation dose of 54 Gy appeared sufficient. Application of postoperative radiotherapy should be decided individually, taking into account the risk of local failure, the need for another craniotomy, and potential radiation toxicity.


Smad2 and PEA3 cooperatively regulate transcription of response gene to complement 32 in TGF-β-induced smooth muscle cell differentiation of neural crest cells.
Response gene to complement 32 (RGC-32) is activated by transforming growth factor- β (TGF-β) and plays an important role in smooth muscle cell (SMC) differentiation from neural crest Monc-1 cells. The molecular mechanism governing TGF-β activation of RGC-32, however, remains to be determined. The present studies indicate that TGF-β regulates RGC-32 gene transcription. Sequence analysis revealed a Smad binding element (SBE) located in the region from -1344 to -1337 bp upstream of the transcription start site of RGC-32 gene. A polyomavirus enhancer activator (PEA3) binding site is adjacent to the SBE. Mutation at either SBE or PEA3 site significantly inhibited RGC-32 promoter activity. Mutations at both sites completely abolished TGF-β-induced promoter activity. Biochemically, TGF-β stimulated recruitment of Smad2, Smad4, and PEA3 to the RGC-32 promoter, as revealed by gel shift and chromatin immunoprecipitation analyses. Functionally, Smad2, but not Smad3, activated RGC-32 promoter. PEA3 appeared to enhance Smad2 activity. In agreement with their function, Smad2, but not Smad3, physically interacted with PEA3. In TGF-β-induced SMC differentiation of Monc-1 cells, knockdown of Smad2 by short hairpin RNA resulted in downregulation of RGC-32 and SMC marker genes. The downregulation of SMC markers, however, was rescued by exogenously introduced RGC-32. These results demonstrate that Smad2 regulation of RGC-32 transcription is essential for SMC differentiation from neural crest cells.


Medical countermeasures for platelet regeneration after radiation exposure. Report of a workshop and guided discussion sponsored by the National Institute of Allergy and Infectious Diseases, Bethesda, MD, March 22–23, 2010.
The events of September 11, 2001 and their aftermath increased awareness of the need to develop medical countermeasures (MCMs) to treat potential health consequences of a radiation accident or deliberate attack. The medical effects of lethal exposures to ionizing radiation have been well described and affect multiple organ systems. To date, much of the research to develop treatments for mitigation of radiation-induced hematopoietic damage has focused on amelioration of radiation-induced neutropenia, which has long been considered to be the primary factor in determining survival after an unintentional radiation exposure. Consistent with historical data, recent studies have highlighted the role that radiation-induced thrombocytopenia plays in radiation mortality, yet development of MCMs to mitigate radiation damage to the megakaryocyte lineage has lagged behind anti-neutropenia approaches. To address this gap and to foster research in the area of platelet regeneration after radiation exposure, the National Institute of Allergy and Infectious Diseases (NIAID) sponsored a workshop on March 22-23, 2010 to encourage collaborations between NIAID program awardees and companies developing pro-platelet approaches. NIAID also organized an informal, open discussion between academic investigators, product development contractors, and representatives from the U.S. Food and Drug Administration (FDA) and other relevant government agencies about drug development toward FDA licensure of products for an acute radiation syndrome indication. Specific emphasis was placed on the challenges of product licensure for radiation/nuclear MCMs using current FDA regulations (21 CFR Parts 314 and 601) and on the importance of animal efficacy model development, design of pivotal protocols, and standardization of irradiation and animal supportive care.


Diabetes alters activation and repression of pro- and anti-inflammatory signaling pathways in the vasculature.
A central mechanism driving vascular disease in diabetes is immune cell-mediated inflammation. In diabetes, enhanced oxidation and glycation of macromolecules, such as lipoproteins, insults the endothelium, and activates both innate and adaptive arms of the immune system by generating new antigens for presentation to adaptive immune cells. Chronic inflammation of the endothelium in diabetes leads to continuous infiltration and accumulation of leukocytes at sites of endothelial cell injury. We will describe the central role of the macrophage as a source of signaling molecules and damaging by-products which activate infiltrating lymphocytes in the tissue and contribute to the pro-oxidant and pro-inflammatory microenvironment. An important aspect to be considered is the diabetes-associated defects in the immune system, such as fewer or dysfunctional athero-protective leukocyte subsets in the diabetic lesion compared to non-diabetic lesions. This review will discuss the key pro-inflammatory signaling pathways responsible for leukocyte recruitment and activation in the injured vessel, with particular focus on pro- and anti-inflammatory pathways aberrantly activated or repressed in diabetes. We aim to describe the interaction between advanced glycation end products and their principle receptor RAGE, angiotensin II, and the Ang II type 1 receptor, in addition to reactive oxygen species (ROS) production by NADPH-oxidase enzymes that are relevant to vascular and immune cell function in the context of diabetic vasculopathy. Furthermore, we will touch on recent advances in epigenetic medicine that have revealed high glucose-mediated changes in the transcription of genes with known pro-inflammatory downstream targets. Finally, novel anti-atherosclerosis strategies that target the vascular immune interface will be explored; such as vaccination against modified low-density lipoprotein and pharmacological inhibition of ROS-producing enzymes.


Learning with interactive computer graphics in the undergraduate neuroscience classroom.
Instruction of neuroanatomy depends on graphical representation and extended self-study. As a consequence, computer-based learning environments that incorporate interactive graphics should facilitate instruction in this area. The present study evaluated such a system in the undergraduate neuroscience classroom. The system used the method of adaptive exploration, in which exploration in a high fidelity graphical environment is integrated with immediate testing and feedback in repeated cycles of learning. The results of this study were that students considered the graphical learning environment to be superior to typical classroom materials used for learning neuroanatomy. Students managed the frequency and duration of study, test, and feedback in an efficient and adaptive manner. For example, the number of tests taken before reaching a minimum test performance of 90 % correct closely approximated the values seen in more regimented experimental studies. There was a wide range of student opinion regarding the choice between a simpler and a more graphically compelling program for learning sectional anatomy. Course outcomes were predicted by individual differences in the use of the software that reflected general work habits of the students, such as the amount of time committed to testing. The results of this introduction into the classroom are highly encouraging for development of computer-based instruction in biomedical disciplines.


Fully integrated digital microfluidics platform for automated immunoassay; A versatile tool for rapid, specific detection of a wide range of pathogens.
With the tangible threat posed by the release of chemical and biological warfare (CBW) agents, detection of airborne pathogens is a critical military and security concern. Recent air sampling techniques developed for biocollection take advantage of Electrowetting on Dielectric (EWOD) to recover material, producing highly concentrated droplet samples. Bespoke EWOD-based digital microfluidics platforms are very well suited to take full advantage of the microlitre concentrated droplet resulting from this recovery process. In this paper we present a free-standing, fully automated DMF platform for immunoassay. Using this system, we demonstrate the automated detection of four classes of CBW agent simulant biomolecules and organisms each representing credible threat agents. Taking advantage of the full magnetic separation process with antibody-bound microbeads, rapid and complete separation of specific target antigen can be achieved with minimal washing steps allowing for very rapid detection. Here, we report clear detection of four categories of antigens achieved with assay completion times of between six and ten minutes. Detection of HSA, Bacillus atrophaeus (BG spores), MS2 bacteriophage and Escherichia coli are demonstrated with estimated limit of detection of respectively 30 ng ml-1, 4 × 104 cfu ml-1, 106 pfu ml-1 and 2 × 107 cfu ml-1. The fully-integrated portable platform described in this paper is highly compatible with the next generation of electrowetting-coupled air samplers and thus shows strong potential toward future in-field deployable biodetection systems and could have key implication in life-changing sectors such as healthcare, environment or food security.


Environmental robustness of the global yeast genetic interaction network.
Phenotypes associated with genetic variants can be altered by interactions with other genetic variants (GxG), with the environment (GxE), or both (GxGxE). Yeast genetic interactions have been mapped on a global scale, but the environmental influence on the plasticity of genetic networks has not been examined systematically. To assess environmental rewiring of genetic networks, we examined 14 diverse conditions and scored 30,000 functionally representative yeast gene pairs for dynamic, differential interactions. Different conditions revealed novel differential interactions, which often uncovered functional connections between distantly related gene pairs. However, the majority of observed genetic interactions remained unchanged in different conditions, suggesting that the global yeast genetic interaction network is robust to environmental perturbation and captures the fundamental functional architecture of a eukaryotic cell.


The role of thymidylate synthase in cellular regulation.
Thymidylate synthase plays a central role in the biosynthesis of thymidylate, an essential precursor for DNA biosynthesis. In addition to its role in catalysis and cellular metabolism, studies from our laboratory have shown that thymidylate synthase functions as an RNA binding protein. Specifically, thymidylate synthase binds with high affinity to its own mRNA resulting in translational repression. An extensive series of experiments have now been performed to elucidate the molecular elements underlying the interaction between thymidylate synthase and its own mRNA. These studies have shed new light into the critical nucleotide sequences and/or secondary structure that are important for protein recognition. As well, studies to define the domains on the protein essential for RNA binding are currently underway. In addition to the characterization of the cis- and trans-acting elements underlying the interaction between thymidylate synthase and its own mRNA, we have recently shown that thymidylate synthase has the capacity to specifically bind in vitro and in vivo to other cellular RNA species. In this regard, thymidylate synthase interacts with the mRNAs of the c-myc onocogene and the p53 tumor suppressor gene. These two genes have been shown to play critical roles in cell cycle control, DNA biosynthesis, and apoptosis. In vitro studies reveal that the interaction of TS with these cell-cycle related mRNAs results in their translational repression. While the biological significance of these cellular RNA/TS protein interactions remains to be defined, these studies suggest a potential role for TS as a mediator in the coordinate regulation of several critical aspects of cellular metabolism.


Regional blood flow and skeletal muscle energy status in endotoxemic rats.
Endotoxins induce muscle wasting in part as a result of depressed protein synthesis. To investigate whether these changes reflect impaired energy transduction, blood flow, O2 extraction, and high-energy phosphates in muscle and whole-body O2 consumption (VO2) have been measured. VO2 was measured for 6h after an initial sublethal dose of endotoxin (Escherichia coli lipopolysaccharide 0.3 mg/100 g body wt sc) or saline and during 6h after a second dose 24 h later. In fed or fasted rats, VO2 was either increased or better maintained after endotoxin. In anesthetized fed rats 3-4 after the second dose of endotoxin VO2 was increased, and this was accompanied by increased blood flow to liver (hepatic arterial supply), kidney, and perirenal brown adipose tissue and a 57 and 64% decrease in flow to back and hindlimb muscle, respectively, with no change in any other organ. Hindlimb arteriovenous O2 was unchanged, indicating markedly decreased aerobic metabolism in muscle, and the contribution of the hindlimb to whole-body VO2 decreased by 46%. Adenosine 5'-triphosphate levels in muscle were unchanged in endotoxin-treated rats, and this was confirmed by topical nuclear magnetic resonance spectroscopy, which also showed muscle pH to be unchanged. These results show that although there is decreased blood flow and aerobic oxidation in muscle, adenosine 5'-triphosphate availability does not appear to be compromised so that the endotoxin-induced muscle catabolism and decreased protein synthesis must reflex some other mechanism.


Temporal changes in function and regional glucose uptake within stunned porcine myocardium.
This study compared the effects of porcine myocardial stunning on the uptake of [18F]-fluorodeoxyglucose (FDG) at 24 hr and 7 days after reperfusion. Prior studies in animals subjected to severe myocardial ischemia have shown a sustained increase in FDG uptake relative to perfusion (FDG/MBF). The time course of recovery of FDG/MBF relative to function poststunning, however, has not been well characterized. Stunning was induced in eight swine by partially occluding the LAD artery for 20 min. At 1 and 7 days postreperfusion, function was assessed by two-dimensional echocardiography and PET studies were obtained with FDG and either 15O-water or 13N-ammonia. Blood flow by microspheres was determined at baseline, during ischemia and after stunning. Myocardial uptake of FDG relative to blood flow on matching images (FDG/MBF) was calculated for all ROIs and expressed as a ratio of LAD to non-LAD areas. After stunning, left ventricular ejection fraction (LVEF) increased from 42% +/- 10% on Day 1 to 52% +/- 6% on Day 7 (p < 0.05). At Day 1, myocardial blood flow was 0.60 +/- 0.10 ml/min/g in LAD and 0.67 +/- 0.16 in non-LAD regions and neither differed at Day 7. The magnitude of FDG/MBF in the LAD region when normalized to the non-LAD region was 1.29 +/- 0.16 on Day 1 and 1.09 +/- 0.08 on Day 7 (p < 0.05) and was inversely proportional to global measures of LVEF (r2 = 0.61; p < 0.005). The severity of postischemic LV dysfunction at 1 and 7 days after stunning correlates with the degree of enhanced regional glucose uptake as estimated by PET. Both normalize within 7 days, suggesting that metabolic and functional abnormalities within completely reperfused myocardium recover in parallel.


Immobilization of Fe-Doped Ni2P Particles Within Biomass Agarose-Derived Porous N,P-Carbon Nanosheets for Efficient Bifunctional Oxygen Electrocatalysis.
A feasible and green sol-gel method is proposed to fabricate well-distributed nano-particulate Fe-Ni2P incorporated in N, P-codoped porous carbon nanosheets (Fe-Ni2P@N,P-CNSs) using biomass agarose as a carbon source, and ethylenediamine tetra (methylenephosphonic acid) (EDTMPA) as both the N and P source. The doped Fe in Ni2P is essential for a substantial increase in intrinsic catalytic activity, while the combined N,P-containing porous carbon matrix with a better degree of graphitization endows the prepared Fe-Ni2P@N,P-CNSs catalyst with a high specific surface area and improved electrical conductivity. Benefiting from the specific chemical composition and designed active site structure, the as-synthesized Fe-Ni2P@N,P-CNSs manifests a satisfying catalytic performance toward both oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) in an alkaline solution, with low overpotential, small Tafel slope and long-term durability, relative to the counterparts (Fe-free Ni12P5/Ni2P2O7@N,P-CNSs and CNSs) with single components and even comparable to Pt/C and RuO2 catalysts. The present work broadens the exploration of efficient bifunctional oxygen electrocatalysts using earth abundant biomass as carbon sources based on non-noble metals for low cost renewable energy conversion/storage.


[What is sustainability science?].
Sustainability is the theme of our time and also the grandest challenge to humanity. Since the 1970s, the term, sustainable development, has frequently appeared in the scientific literature, governmental documents, media promotions for public goods, and commercial advertisements. However, the science that provides the theoretical foundation and practical guidance for sustainable development--sustainability science--only began to emerge in the beginning of the 21st century. Nevertheless, the field has rapidly developed in depth and expanded in scope during the past decade, with its core concepts and research methods coalescing. China, as the most populous country in the world and home to the philosophical root of sustainability science-the unity of man and nature, is obligated to take upon the challenge of our time, to facilitate global sustainability while pursuing the Chinese Dream, and to play a leading role in the development of sustainability science. Toward this grandiose goal, this paper presents the first Chinese introduction to sustainability science, which discusses its basic concepts, research questions, and future directions. Sustainability science is the study of the dynamic relationship between humans and the environment, particularly focusing on the vulnerability, robustness, resilience, and stability of the coupled human-environment system. It is a transdisciplinary science that integrates natural sciences with humanities and social sciences. It hinges on the environment-economy-society nexus, and merges basic and applied research. The key components of sustainability often change with time, place, and culture, and thus sustainability science needs to emphasize multi-scale studies in space and time, with emphasis on landscapes and regions over a horizon of 50 to 100 years. It needs to focus on the relationship between ecosystem services and human well-being, as influenced by biodiversity and ecosystem processes as well as climate change, land use change, and other socioeconomic drivers. Landscape sustainability is at the core of sustainability science, and is expected to be a hot research topic in the next few decades.


Conjugation of palmitic acid improves potency and longevity of siRNA delivered via endosomolytic polymer nanoparticles.
Clinical translation of siRNA therapeutics has been limited by the inability to effectively overcome the rigorous delivery barriers associated with intracellular-acting biologics. Here, to address both potency and longevity of siRNA gene silencing, pH-responsive micellar nanoparticle (NP) carriers loaded with siRNA conjugated to palmitic acid (siRNA-PA) were investigated as a combined approach to improve siRNA endosomal escape and stability. Conjugation to hydrophobic PA improved NP loading efficiency relative to unmodified siRNA, enabling complete packaging of siRNA-PA at a lower polymer:siRNA ratio. PA conjugation also increased intracellular uptake of the nucleic acid cargo by 35-fold and produced a 3.1-fold increase in intracellular half-life. The higher uptake and improved retention of siRNA-PA NPs correlated to a 2- and 11-fold decrease in gene silencing IC50 in comparison to siRNA NPs in fibroblasts and mesenchymal stem cells, respectively, for both the model gene luciferase and the therapeutically relevant gene prolyl hydroxylase domain protein 2 (PHD2) . PA conjugation also significantly increased longevity of silencing activity following a single treatment in fibroblasts. Thus, conjugation of PA to siRNA paired with endosomolytic NPs is a promising approach to enhance the functional efficacy of siRNA in tissue regenerative and other applications.


Endogenous interleukin (IL)-17A promotes pristane-induced systemic autoimmunity and lupus nephritis induced by pristane.
Interleukin (IL)-17A is increased both in serum and in kidney biopsies from patients with lupus nephritis, but direct evidence of pathogenicity is less well established. Administration of pristane to genetically intact mice results in the production of autoantibodies and proliferative glomerulonephritis, resembling human lupus nephritis. These studies sought to define the role of IL-17A in experimental lupus induced by pristane administration. Pristane was administered to wild-type (WT) and IL-17A(-/-) mice. Local and systemic immune responses were assessed after 6 days and 8 weeks, and autoimmunity, glomerular inflammation and renal injury were measured at 7 months. IL-17A production increased significantly 6 days after pristane injection, with innate immune cells, neutrophils (Ly6G(+)) and macrophages (F4/80(+)) being the predominant source of IL-17A. After 8 weeks, while systemic IL-17A was still readily detected in WT mice, the levels of proinflammatory cytokines, interferon (IFN)-γ and tumour necrosis factor (TNF) were diminished in the absence of endogenous IL-17A. Seven months after pristane treatment humoral autoimmunity was diminished in the absence of IL-17A, with decreased levels of immunoglobulin (Ig)G and anti-dsDNA antibodies. Renal inflammation and injury was less in the absence of IL-17A. Compared to WT mice, glomerular IgG, complement deposition, glomerular CD4(+) T cells and intrarenal expression of T helper type 1 (Th1)-associated proinflammatory mediators were decreased in IL-17A(-/-) mice. WT mice developed progressive proteinuria, but functional and histological renal injury was attenuated in the absence of IL-17A. Therefore, IL-17A is required for the full development of autoimmunity and lupus nephritis in experimental SLE, and early in the development of autoimmunity, innate immune cells produce IL-17A.


Coding of modified body schema during tool use by macaque postcentral neurones.
A tool is an extension of the hand in both a physical and a perceptual sense. The presence of body schemata has been postulated as the basis of the perceptual assimilation of tool and hand. We trained macaque monkeys to retrieve distant objects using a rake, and neuronal activity was recorded in the caudal postcentral gyrus where the somatosensory and visual signals converge. There we found a large number of bimodal neurones which appeared to code the schema of the hand. During tool use, their visual receptive fields were altered to include the entire length of the rake or to cover the expanded accessible space. These findings may represent neural correlates of the modified schema of the hand in which the tool was incorporated.


Phenotypic similarities and differences between CALLA-positive acute lymphoblastic leukemia cells and normal marrow CALLA-positive B cell precursors.
Expression of differentiation markers in common acute lymphoblastic leukemia (cALL) cells from 25 patients was compared with subpopulations of normal common ALL antigen (CALLA) (CD10)-positive bone marrow lymphoid cells (cBMLs). In cBML, CD10 intensity is positively correlated with CD34 (MY10) and terminal transferase (TdT) expression and inversely correlated with common leukocyte antigen (CLA), CD20 (B1), and ctyoplasmic mu chain (Cmu) expression. In cALL, CD10 density was inversely correlated with CLA and Cmu expression and strongly correlated with CD34 expression as in cBML. In contrast to cBML, TdT and CD20 expression were not related to CD10 density in cALL. Furthermore, cALL TdT intensity measured by enzyme immunoassay was not related to expression of CD10, CLA, or CD34, but was positively correlated with CD20 expression. Cmu expression in cALL was inversely correlated with expression of CD34 and positively correlated with CLA as in cBML, but showed no association with TdT intensity or CD20 expression in contrast to the relationship found in cBMLs. Analysis of TdT intensity and Cmu expression in sorted subpopulations of cells from individual patients that were positive or negative for CD34, CD20, or CD10 was consistent with the data obtained by comparison of cells from different patients. These results indicate that from patient to patient and within individual patients, cALL cells express the markers CD34, CLA, CD10, and Cmu in a coordinated fashion similar to cBMLs, but demonstrate differences in expression of TdT and CD20 with respect to the marrow cells considered their normal counterparts. The cALL cells that are CD34 positive show increased expression of CD10 and are less likely to be CLA or Cmu positive, suggesting that they may represent a phenotypically less differentiated form of cALL than does CD34-negative cALL.


[Detection of noncapsular antigens in pneumococcus].
Complex antigenic preparations obtained from noncapsular pneumococcal strains were used for the immunization of rabbits and guinea pigs. The injection of the preparations in complete Freund's adjuvant for 5 weeks led to the appearance of antibodies in their sera. The antibodies were detected by the double immunodiffusion test. The preparations obtained from different strains by extraction (with Triton X-100 or sodium deoxycholate) or by disintegration contain common pneumococcal antigens.


Changes in readiness to quit and self-efficacy among adolescents receiving a brief office intervention for smoking cessation.
To examine changes in readiness to quit and self-efficacy among adolescents who received a clinic-based, brief office intervention (BOI) for smoking cessation. This study utilized a prospective, pre-post- treatment design. Participants were adolescent smokers (34 females, 35 males) with a mean +/- SD age of 15.8 +/- 1.4 years; 86% were Caucasian, who were randomly assigned to receive the BOI as part of a larger clinical trial. They were recruited from three cities in the Midwest and Northeastern part of the United States. After the baseline assessment, the BOI was designed for adolescents to receive four weekly individual sessions with a research counselor lasting between 10 and 40 min each. The BOI includes motivational interviewing and cognitive-behavioral techniques. Readiness to quit was assessed at each treatment session using the stages of change algorithm. The validated Adolescent Smoking Self-Efficacy Scale (SES) was used to assess self-efficacy at baseline (week 0) prior to the intervention and at post-treatment (week 4). The SES items comprise three factors or subscales: opportunities to smoke, emotional stress, and friends' influence. The percentage of adolescents who made improvement on readiness to quit from the baseline treatment session was statistically significant (p < .001) for each of the three subsequent treatment sessions. Self-efficacy scores increased significantly (p < .004) from baseline to post-treatment for all three subscales. Adolescents receiving a BOI progressed in their readiness and self-efficacy to quit. Understanding the change process among adolescent smokers during treatment could influence the design of future stop smoking interventions.


Evaluation of vocal changes after vestibular rehabilitation in patients with balance disorders.
Although there are articles and studies that associate postural changes with changes in vocal quality, to the best of our knowledge, this was the first study investigating the association between balance disorders and voice. This study aimed to determine whether patients with balance disorders present any clinical, acoustic, or endoscopic vocal changes, and if the correction of balance impairments, such as through vestibular rehabilitation, lead to improvement in vocal quality. This was a prospective cohort study that analyzed vocal differences (clinical, videoendostroboscopic, audio-perceptual, and acoustic vocal parameters) in a sample of 43 patients with vestibular dysfunction at three different time points (pre-treatment, post-treatment, and 3 months' post-treatment) diagnosed by videonystagmography with changes in computerized dynamic posturography who were treated with vestibular rehabilitation. In pre-treatment, all of the patients presented supraglottic hyperfunction during videoendoscopic examination and abnormal values in the audio-perceptual scale. After treatment for balance disorders, there was a statistically significant improvement in some parameters of the videoendoscopic and audio-perceptual measures. These improvements were detected immediately after treatment and remained present until at least three months after treatment. The results suggested that the treatment for balance disorders results in changes in posture and consequently in voice quality.


c-KIT gene mutation and clonal origin of multiple gastrointestinal stromal tumors occurring in the peritoneum.
The present study investigated multiple extragastrointestinal stromal tumors (EGISTs) occurring in the peritoneum, identified their clinicopathologic and immunohistochemical features, and examined their c-KIT gene mutations, clonal status and clonal relationships. Fifteen different tumors of a multiple EGIST from the same female patient were examined by microscopy and immunohistochemistry, then genomic DNA was isolated from the lesions and, as the control, the surrounding fibrous connective tissues. PCR amplification and sequencing were used to investigate the mutation status of c-KIT gene exons 9, 11, 13 and 17. Finally, a clonality assay was conducted based on X-chromosome inactivation mosaicism in female somatic tissues and polymorphisms of the phosphoglycerate kinase and androgen receptor genes. Immunohistochemical analysis revealed CD117- and CD34-positive reactivity in the tumors. Mutation analysis identified the c-KIT gene mutation in exon 13. The clonality assay revealed a loss of X-chromosome inactivation mosaicism and an identical inactivated allele in all 15 nodules. No similar findings were observed in the surrounding fibrous connective tissues. The c-KIT gene mutation was found in the multiple EGISTs from the patient. The EGIST, like most tumors, was of monoclonal origin. This and the identical inactivated allele found in the nodules indicated a common clonal origin for the tumor.


A Disulfide-Stabilized Aβ that Forms Dimers but Does Not Form Fibrils.
Aβ dimers are a basic building block of many larger Aβ oligomers and are among the most neurotoxic and pathologically relevant species in Alzheimer's disease. Homogeneous Aβ dimers are difficult to prepare, characterize, and study because Aβ forms heterogeneous mixtures of oligomers that vary in size and can rapidly aggregate into more stable fibrils. This paper introduces AβC18C33 as a disulfide-stabilized analogue of Aβ42 that forms stable homogeneous dimers in lipid environments but does not aggregate to form insoluble fibrils. The AβC18C33 peptide is readily expressed in Escherichia coli and purified by reverse-phase HPLC to give ca. 8 mg of pure peptide per liter of bacterial culture. SDS-PAGE establishes that AβC18C33 forms homogeneous dimers in the membrane-like environment of SDS and that conformational stabilization of the peptide with a disulfide bond prevents the formation of heterogeneous mixtures of oligomers. Mass spectrometric (MS) studies in the presence of dodecyl maltoside (DDM) further confirm the formation of stable noncovalent dimers. Circular dichroism (CD) spectroscopy establishes that AβC18C33 adopts a β-sheet conformation in detergent solutions and supports a model in which the intramolecular disulfide bond induces β-hairpin folding and dimer formation in lipid environments. Thioflavin T (ThT) fluorescence assays and transmission electron microscopy (TEM) studies indicate that AβC18C33 does not undergo fibril formation in aqueous buffer solutions and demonstrate that the intramolecular disulfide bond prevents fibril formation. The recently published NMR structure of an Aβ42 tetramer (PDB: 6RHY) provides a working model for the AβC18C33 dimer, in which two β-hairpins assemble through hydrogen bonding to form a four-stranded antiparallel β-sheet. It is anticipated that AβC18C33 will serve as a stable, nonfibrilizing, and noncovalent Aβ dimer model for amyloid and Alzheimer's disease research.


Distribution of de novo synthesized betaine in rat kidney: role of renal synthesis on medullary betaine accumulation.
The trimethylamine glycine-betaine is accumulated to high concentrations in medullary cells of mammalian kidneys, whereas betaine synthesis from choline is predominant in the renal cortex. We investigated the contribution of renal betaine synthesis to medullary betaine accumulation. De novo synthesis of betaine in situ was accomplished by injecting [14C]choline into the renal artery of male Sprague-Dawley rats. [14C]betaine was measured in the renal cortex and medulla, as well as in serum and urine samples. Betaine concentration in the cortex decreased from 3.5 +/- 1.3 at 5 min to 0.4 +/- 0.2 nmol/mg protein at 60 min, but it increased from 1.4 +/- 0.1 to 2.5 +/- 0.6 nmol/mg protein in the medulla. Serum and total urine [14C]betaine increased from 2.7 +/- 1.3 and 0.9 +/- 0.1 nmol/ml at 5 min to 5.3 +/- 0.3 and 2.1 +/- 0.4 nmol/ml at 60 min, respectively. Concentrations of newly synthesized betaine were not decreased by the ligation of the hepatic artery and portal vein, suggesting that most [14C]betaine was synthesized in the kidney. Coinjection with 5 mM dimethylamino-ethanol, a choline oxidase inhibitor, and 100 mM cold betaine reduced medullary betaine accumulation by 80 and 76%, respectively. Water deprivation for 60 h increased both cortical and medullary [14C]betaine, whereas furosemide diuresis decreased the medullary [14C]betaine concentration. We concluded that betaine synthesized in the kidney can be accumulated in the medulla and that the medullary concentrations of newly synthesized betaine are closely related to the hydration state of the animal.


FDG PET/CT findings in recurrent malignant schwannoma.
Malignant schwannoma is an uncommon but aggressive sarcoma that most commonly arises in young and middle-aged adults. We present a 28-year-old male patient with a recurrent chest wall malignant schwannoma. An FDG PET/CT was performed to evaluate the management of the patient. The PET/CT images showed hypermetabolic mass lesions in the left upper chest wall and increased FDG uptake in the left axillary region. The pathologic results confirmed the PET findings. We conclude that FDG PET/CT could play an important role in the staging, restaging, and post-therapy follow-up of malignant schwannomas.


Voluntary modulation of the vestibuloocular reflex in humans and its relation to smooth pursuit.
Human subjects attempted to modify their vestibuloocular reflex (VOR) in the dark by fixating imagined targets while experiencing predictable (SIN) sinusoidal (0.01-2.5 Hz) and unpredictable (SSN) sum of sines rotational stimuli (0.02-1.9 Hz). Modification was attempted under 2 instructional sets: VOR enhancement, ie tracking an imaginary earth-fixed target; VOR suppression, ie fixation of a chair fixed target. When compared to gain characteristics exhibited during the relax state with the same stimuli, subjects were able to alter VOR gain under both experimental conditions, raising it during the enhance paradigm and lowering it during the suppress paradigm. While ability to suppress the VOR was dependent on stimulus frequency, decreasing as frequency of rotation increased, subjects were equally able to modify their responses to the unpredictable and the predictable stimuli. Response phase did not change and was maintained close to 180 deg, regardless of instructional set, predictability, or frequency of stimulation for frequencies greater than 0.1 Hz. At frequencies below 0.1 Hz, a phase lead developed that was similar for all paradigms and rotational stimuli. In contrast, when subjects attempted to pursue visual targets that matched closely the velocities and frequencies of the chair rotation during predictable (SIN) and unpredictable stimulation (SSN), success was dependent on predictability of the stimulus. SSN target motion caused a significant decrease in pursuit velocity as compared to results using SIN target motion. Phase characteristics for both types of stimuli were similar, demonstrating a slight lead at lower frequencies and lagging as frequency of target oscillation increased. The results suggest that voluntary modulation of the VOR is not mediated by a neural control mechanism that is based on prediction. In addition, pursuit does not appear to contribute significantly to ability to cancel VOR. Instead, VOR modulation may be a cognitive event that involves use of a mechanism that produces simple parametric gain changes.


Fascicle specific targeting for selective peripheral nerve stimulation.
Electrical stimulation is a blunt tool for evoking neural activity. Neurons are naturally activated asynchronously and non-uniformly, whereas stimulation drives simultaneous activity within a population of cells. These differences in activation pattern can result in unintended side effects, including muddled sensory percepts and undesirable muscle contractions. These effects can be mitigated by the placement of electrodes in close approximation to nerve fibers and careful selection of the neural interface's location. This work describes the benefits of placing electrodes within specific fascicles of peripheral nerve to form selective neural interfaces for bidirectional neuroprosthetic devices. Chronic electrodes were targeted to individual fascicles of the ulnar and median nerves in the forearm of four human subjects. During the surgical implant procedure, fascicles were dissected from each nerve, and functional testing was used to identify the relative composition of sensory and motor fibers within each. FAST-LIFE arrays, composed of longitudinal intrafascicular arrays and fascicular cuff electrodes, were implanted in each fascicle. The location, quality, and stimulation parameters associated with sensations evoked by electrical stimulation on these electrodes were characterized throughout the 90-180 d implant period. FAST-LIFE arrays enable selective and chronic electrical stimulation of individual peripheral nerve fascicles. The quality of sensations evoked by stimulation in each fascicle is predictable and distinct; subjects reported tactile and cutaneous sensations during stimulation of sensory fascicles and deeper proprioceptive sensations during stimulation of motor fascicles. Stimulation thresholds and strength-duration time constants were typically higher within sensory fascicles. Highly selective, stable neural interfaces can be created by placing electrodes within and around single fascicles of peripheral nerves. This method enables targeting electrodes to nerve fibers that innervate a specific body region or have specific functions. Fascicle-specific interfacing techniques have broad potential to maximize the therapeutic effects of electrical stimulation in many neuromodulation applications. (Clinical Trial ID NCT02994160.).


Dysbiosis, Spleen Qi, Phlegm, and Complex Difficulties.
The incidence of complex chronic health conditions is rising. Some of these conditions might have their roots in gut disturbances, which in Chinese Medicine would be categorized as Spleen and Stomach Qi Disharmonies. In this article, the author explores these issues from an integrative perspective and suggests areas where acupuncture techniques could be helpful. Five cases are described briefly to illustrate the use of acupuncture.


Cytochrome P450 Monooxygenase CYP716A141 is a Unique β-Amyrin C-16β Oxidase Involved in Triterpenoid Saponin Biosynthesis in Platycodon grandiflorus.
The roots of Platycodon grandiflorus are widely used as a crude drug. The active components include a variety of triterpenoid saponins. Recent studies have revealed that Cyt P450 monooxygenases (P450s) function as triterpene oxidases in triterpenoid saponin biosynthesis in many plant species. However, there have been no reports regarding triterpene oxidases in P. grandiflorus. In this study, we performed transcriptome analysis of three different P. grandiflorus tissues (roots, leaves and petals) using RNA sequencing (RNA-Seq) technology. We cloned six P450 genes that were highly expressed in roots, and classified them as belonging to the CYP716A, CYP716D and CYP72A subfamilies. We heterologously expressed these P450s in an engineered yeast strain that produces β-amyrin, one of the most common triterpenes in plants. Two of the CYP716A subfamily P450s catalyzed oxidation reactions of the β-amyrin skeleton. One of these P450s, CYP716A140v2, catalyzed a three-step oxidation reaction at C-28 on β-amyrin to produce oleanolic acid, a reaction performed by CYP716A subfamily P450s in a variety of plant species. The other P450, CYP716A141, catalyzed the hydroxylation of β-amyrin at C-16β. This reaction is unique among triterpene oxidases isolated to date. These results enhance our knowledge of functional variation among CYP716A subfamily enzymes involved in triterpenoid biosynthesis, and provide novel molecular tools for use in synthetic biology to produce triterpenoid saponins with pre-defined structures.


Challenges to the Implementation of Interprofessional Education in Health Profession Education in Iran.
Changes in the structure and nature of health care highlight the need for better collaboration between professions focusing on patient or health-care users. Interprofessional education (IPE) has attracted a lot of attention at the international level, and its positive consequences in different areas of health care have been investigated and approved. This study explored the opinions and viewpoints of professors and experts regarding the challenges to the implementation of interprofessional education in health profession education in Iran in 2018. This qualitative study was conducted with 15 IPE professors and experts using semi-structured interviews. We used an inductive approach to content analysis as developed by Elo et al. The interviews were analyzed using Graneheim and Lundman method by MAXQDA software version 12. The main challenges to the implementation of IPE were categorized into educational system, structural and cultural challenges. The educational challenges were the professors' lack of ability, a non-flexible and single-disciplined curriculum, and weaknesses in the current education. The professors and experts felt that the most important structural challenges included centralism, hierarchy of power, poor educational groundwork, and lack of support of organizations. Cultural challenges mainly related to the attitudes of managers, professors, and even students regarding IPE and the need for change. Designing and implementing IPE in Iran face major challenges. Cultural and attitude issues were the greatest challenges to be addressed in the context of implementing IPE in health-care education.


Recommendations to increase the impact of maternal and childbirth health systematic reviews in the Americas.
This paper summarises the discussions and the recommendations formulated during a meeting in March 2007 on the challenges and strategies to increase the impact of maternal and childbirth health systematic reviews in the Americas. The discussions addressed three specific themes: (1) performing systematic reviews (2) updating existing reviews, and (3) diffusing and implementing evidence into practice. Practical recommendations were devised for each theme in small group discussions.


Crystal structure of the Rho-associated coiled-coil kinase 2 inhibitor belumosudil bound to CK2α.
The small molecule belumosudil was initially identified as a selective inhibitor of Rho-associated coiled-coil kinase 2 (ROCK2) and has recently been approved for the treatment of graft-versus-host disease. However, recent studies have shown that many of the phenotypes displayed upon treatment with belumosudil were due to CK2α inhibition. CK2α is in itself a very promising therapeutic target for a range of conditions and has recently been put forward as a potential treatment for COVID-19. Belumosudil presents a promising starting point for the development of future CK2α inhibitors as it provides a safe, potent and orally bioavailable scaffold. Therefore, several of the major hurdles in drug development have already been overcome. Here, the crystal structure of belumosudil bound to the ATP site of CK2α is presented. This crystal structure combined with modelling studies further elucidates how belumosudil could be developed into a selective and potent CK2α or ROCK2 inhibitor.


Anthracosis mimicking mediastinal lymph node metastases with 18F-FCholine in high-risk prostate cancer.
A 62-year-old patient with prostate adenocarcinoma underwent PET with radiolabeled choline (18F-Fcholine) for pretreatment staging of a high-risk prostate cancer. Images showed a significant mediastinal lymph node uptake of 18F-Fcholine. Owing to the rarity of spread to supradiaphragmatic lymph nodes, a surgical removal was performed, revealing anthracosis and no malignant cells. Even if its specificity seems better than 18F-FDG, false positives have been reported and other pathologies could mimic lymph node metastases. Consequently, histology should be performed so that the appropriate treatment can be initiated.


Efficacy of Chloroquine and Primaquine for the Treatment of Uncomplicated Plasmodium vivax Malaria in Cruzeiro do Sul, Brazil.
We evaluated the efficacy of chloroquine and primaquine on uncomplicated Plasmodium vivax malaria in Cruzeiro do Sul, Brazil, in 2014. Patients ≥ 5 years of age with either fever or history of fever, and laboratory-confirmed P. vivax monoinfection received chloroquine (total dose = 25 mg/kg) and primaquine (total dose = 3.5 mg/kg), and were followed up for 168 days (24 weeks). We used microsatellite genotyping to differentiate recurrent infections caused by heterologous parasites from those caused by homologous ones. No new P. vivax episode occurred by Day 28 among 119 enrolled patients, leading to Day 28, with adequate clinical and parasitological response (ACPR) of 100% (95% confidence interval [CI] = 96.7-100%). Twenty-eight P. vivax episodes occurred by Day 168, with uncorrected ACPR of 69.9% (95% CI = 59.5-79.0%). Fifteen of these episodes were caused by either homologous haplotypes or haplotypes that could not be determined. Excluding the 13 recurrent episodes caused by heterologous parasites, Day 168 microsatellite-corrected ACPR was estimated at 81.2% (95% CI = 71.0-89.1%). Chloroquine and primaquine remain efficacious to treat acute uncomplicated P. vivax infection, but moderate recurrence rates were observed within 24 weeks of follow-up.


[Selective 12-lipoxygenase inhibition potentiates the effect of radiation on human prostate cancer cells in vitro and in vivo].
Prostate cancer is one of the leading cancer types in males in the developed world. Radiotherapy is a major method in the curative treatment of prostate cancer however, up to 30% of the patients experience local relapse. Arachidonic acid metabolites have been shown to have important role in cancer. 12-lipoxygenase (12-LOX) has been proven to significantly influence prostate cancer progression, by apoptosis regulation and by promoting cancer cell survival. In this study we examined whether 12-LOX inhibition may increase radiation sensitivity of prostate cancer cells in vitro and in vivo. Prostate cancer cell lines were treated with 12-LOX inhibitors, different doses of radiation and the combination of 12-LOX inhibitors and radiation. We measured the effect of these treatments through clonogenic survival and apoptosis in vitro and tumor growth in vivo in a tumor xenograft model. 12-LOX inhibition and radiation both increased apoptosis and decreased clonogenic survival of prostate cancer cell lines in vitro. Combined treatment resulted in a supra-additive effect in vitro. In vivo both 12-LOX inhibition and radiotherapy caused delay in growth of the xenograft tumors but the combined treatment resulted in the strongest growth inhibition. The presented data prove that 12-LOX and its metabolite 12(S)-HETE have a major role in prostate cancer cell progression and radiosensitivity. We have shown by different methods in vitro and in vivo that inhibition of 12-LOX activity significantly sensitizes prostate cancer cells to radiation. Therefore we can state that 12-LOX inhibitors are promising compounds to be developed to become a new class of clinical radiation sensitizers in prostate cancer.


[Simultaneous operation on the tumor of the pancreas head and abdominal aortic aneurysm-case study].
A case of 73 years old patient qualified for operative treatment due to the escalation of jaundice in the course of the tumor of the pancreas head was presented. During laparotomy, apart from resectable tumor of the pancreas head, abdominal aortic aneurysm of the size 45 x 55 mm was found in a section below renal arteries, not involving iliac arteries. The resection of the abdominal aortic aneurysm and pancreatoduodenectomy were performed simultaneously. The patient was able to leave the hospital on the 12th day after the operation in a good general condition.


Macular morphometrics in foveal displacement following full thickness macular hole surgery.
To investigate if the ratio of the preoperative nasal and temporal "arms" of the macular hole (MH) can have a predictive value in the magnitude of foveal displacement postoperatively. This is retrospective interventional case series of eyes of 40 patients with full-thickness macular hole (FTMH), which underwent vitrectomy with internal limiting membrane (ILM) peeling and had Type 1 closure.All subjects underwent pre and postoperative optical coherence tomography (OCT, Heidelberg, Spectralis, Germany). Their pre and postoperative foveo-papillary distance (FPD) was measured and the magnitude of shift was calculated. The nasal and temporal arm lengths, their ratio (N/T ratio), and the hole base diameter were measured in the preoperative OCTs.The main outcome measure was the correlation of the N/T ratio with the postoperative foveal displacement. We observed that in 25% (n = 10) the fovea shifted temporally; in 75% (n = 30) it shifted nasally. The shift did not show a significant correlation with either N/T ratio (r = 0.155, P = 0.34) or with base diameter (r = -0.008, P = 0.961). The odds ratio (OR) was 4.92 (P = 0.04) and the relative risk (RR) was 3.12 (P = 0.039) for a longer temporal segment to predict a temporal shift. Both nasal and temporal shifts are possible after successful hole closure and temporal shifts can also occur in a significantly high proportion of patients. Temporal shifts are more likely in eyes with a longer temporal segment.


Computer-mediated and face-to-face communication in metastatic cancer support groups.
To compare the experiences of women with metastatic breast cancer (MBC) in computer-mediated and face-to-face support groups. Interviews from 18 women with MBC, who were currently in computer-mediated support groups (CMSGs), were examined using interpretative phenomenological analysis. The CMSGs were in an asynchronous mailing list format; women communicated exclusively via email. All the women were also, or had previously been, in a face-to-face support group (FTFG). CMSGs had both advantages and drawbacks, relative to face-to-face groups (FTFGs), for this population. Themes examined included convenience, level of support, intimacy, ease of expression, range of information, and dealing with debilitation and dying. CMSGs may provide a sense of control and a greater level of support. Intimacy may take longer to develop in a CMSG, but women may have more opportunities to get to know each other. CMSGs may be helpful while adjusting to a diagnosis of MBC, because women can receive support without being overwhelmed by physical evidence of disability in others or exposure to discussions about dying before they are ready. However, the absence of nonverbal cues in CMSGs also led to avoidance of topics related to death and dying when women were ready to face them. Agendas for discussion, the presence of a facilitator or more time in CMSGs may attenuate this problem. The findings were discussed in light of prevailing research and theories about computer-mediated communication. They have implications for designing CMSGs for this population.


Taenia saginata a rare cause of acute cholangitis: a case report.
Parasitic infestations of the galdbladder and biliary tract are quite rare. Taenia saginata is an intestinal helmint and patients harbouring adult T.saginata tapeworms are mostly asymptomatic and discharge only fecal proglottids. In some cases there might be nonspecific symptoms like vomiting, nausea, epigastric pain, diarrhea and weight loss. Tenia saginata is a also rare cause of ileus, pancreatitis, cholecystitis and cholangitis. We report a case of acute cholangitis caused by T. saginata presenting with fever, nausea, vomiting, jaundice and right upper quadrant pain. Although parasites are not an uncommon cause of cholangitis especially in diseaseendemic areas like the Far East, this is not true for T. saginata causing acute cholangitis.


The impact of diabetes mellitus on the prognostic value of a normal dobutamine stress echocardiogram in patients with intermediate to high cardiovascular risk.
There is currently limited data on the prognostic value of a normal dobutamine stress echocardiogram (DSE) in patients with intermediate to high cardiovascular risk. The impact of diabetes mellitus, recently recognised as a cardiovascular risk-equivalent, has not been previously evaluated. This study aims to determine the prognostic value of a normal DSE in these patients. The study population includes all patients with two cardiovascular risk factors or diabetes mellitus and a normal DSE (baseline and peak stress) with three months follow-up. A total of 122 patients (47 females, 75 males; mean age 59.6 years) were recruited. Impact of diabetes mellitus on subsequent cardiovascular events was determined. Diabetes mellitus was present in 32.8 percent, hypertension in 72.1 percent, smoking in 27.0 percent, family history of premature coronary artery disease in 15.6 percent, and hypercholesterolemia in 66.4 percent. On follow-up until 6.4 years (mean 4.1 years), there were four myocardial infarctions (0.8 percent per patient/year) and five revascularisation procedures (1.0 percent per patient/year). The majority of adverse events occurred among patients with diabetes mellitus (three out of four myocardial infarctions; four out of five revascularisations). Diabetes mellitus independently predicted subsequent cardiac events on both univariate and multivariate analyses (p value is equal to 0.015 and 0.011, respectively). Presence of diabetes mellitus also conferred a worse outcome on survival analysis (p value is equivalent to 0.0046). The presence of diabetes mellitus adversely affects clinical outcome despite a normal DSE. Patients without diabetes mellitus, but with intermediate to high cardiovascular risk, and a normal DSE have a better medium term outcome.


Src family tyrosine kinase Lyn mediates VWF/GPIb-IX-induced platelet activation via the cGMP signaling pathway.
The platelet receptor for von Willebrand factor (VWF), glycoprotein (GP) Ib-IX, mediates initial platelet adhesion and transmits signals leading to platelet activation. Src family tyrosine kinases (SFKs) play an important role in VWF-induced GPIb-IX signaling. However, the SFK-dependent downstream signaling pathway is unclear but is thought to involve thromboxane A2 (TXA2) synthesis. Here we show that, although platelets deficient in SFK members, Lyn or Fyn, were defective in the TXA2-dependent second wave of platelet aggregation induced by botrocetin/VWF, only Lyn-knockout platelets were also defective in stable platelet adhesion to VWF under shear stress that is independent of the TXA2 pathway. Lyn-knockout platelets also spread poorly on VWF but spread normally on fibrinogen, indicating an important role for Lyn in VWF-mediated GPIb signaling but not in integrin outside-in signaling. Importantly, Lyn knockout abrogated VWF-induced cGMP elevation. Addition of low concentrations of 8-bromo-cGMP, however, corrected the defective stable adhesion of Lyn-knockout platelets or PP2-treated platelets on VWF. These results demonstrate an important role for Lyn in VWF/GPIb-IX-induced integrin activation mediated via the cGMP signaling pathway independently of TXA2 synthesis and also indicate that Lyn is critically important in GPIb-IX-mediated activation of the cGMP pathway.


18F-FDG Uptake in Parsonage-Turner Syndrome.
A 55-year-old man with large B-cell lymphoma developed atraumatic left shoulder pain. F-FDG PET/CT revealed new left supraspinatus and infraspinatus muscle uptake while the initial disease resolved. Given the discrepancy between initial disease treatment response and new left shoulder findings, an MRI scan was performed. This demonstrated diffuse supraspinatus and infraspinatus muscle edema and enhancement with no focal lesion. Muscle biopsy was negative for lymphoma, but features of muscle denervation were seen. Overall, clinical and imaging findings were compatible with Parsonage-Turner syndrome (acute brachial neuritis), an uncommon condition that presented as a false-positive finding on PET/CT.


Resolving dynamics and function of transient states in single enzyme molecules.
We use a hybrid fluorescence spectroscopic toolkit to monitor T4 Lysozyme (T4L) in action by unraveling the kinetic and dynamic interplay of the conformational states. In particular, by combining single-molecule and ensemble multiparameter fluorescence detection, EPR spectroscopy, mutagenesis, and FRET-positioning and screening, and other biochemical and biophysical tools, we characterize three short-lived conformational states over the ns-ms timescale. The use of 33 FRET-derived distance sets, to screen available T4L structures, reveal that T4L in solution mainly adopts the known open and closed states in exchange at 4 µs. A newly found minor state, undisclosed by, at present, more than 500 crystal structures of T4L and sampled at 230 µs, may be actively involved in the product release step in catalysis. The presented fluorescence spectroscopic toolkit will likely accelerate the development of dynamic structural biology by identifying transient conformational states that are highly abundant in biology and critical in enzymatic reactions.


An unconventional hexacoordinated flavohemoglobin from Mycobacterium tuberculosis.
Being an obligate aerobe, Mycobacterium tuberculosis faces a number of energetic challenges when it encounters hypoxia and environmental stress during intracellular infection. Consequently, it has evolved innovative strategies to cope with these unfavorable conditions. Here, we report a novel flavohemoglobin (MtbFHb) from M. tuberculosis that exhibits unique features within its heme and reductase domains distinct from conventional FHbs, including the absence of the characteristic hydrogen bonding interactions within the proximal heme pocket and mutations in the FAD and NADH binding regions of the reductase domain. In contrast to conventional FHbs, it has a hexacoordinate low-spin heme with a proximal histidine ligand lacking imidazolate character and a distal heme pocket with a relatively low electrostatic potential. Additionally, MtbFHb carries a new FAD binding site in its reductase domain similar to that of D-lactate dehydrogenase (D-LDH). When overexpressed in Escherichia coli or Mycobacterium smegmatis, MtbFHb remained associated with the cell membrane and exhibited D-lactate:phenazine methosulfate reductase activity and oxidized D-lactate into pyruvate by converting the heme iron from Fe(3+) to Fe(2+) in a FAD-dependent manner, indicating electron transfer from D-lactate to the heme via FAD cofactor. Under oxidative stress, MtbFHb-expressing cells exhibited growth advantage with reduced levels of lipid peroxidation. Given the fact that D-lactate is a byproduct of lipid peroxidation and that M. tuberculosis lacks the gene encoding D-LDH, we propose that the novel D-lactate metabolizing activity of MtbFHb uniquely equips M. tuberculosis to balance the stress level by protecting the cell membrane from oxidative damage via cycling between the Fe(3+)/Fe(2+) redox states.


Alcohol Use and Cancers of the Gastrointestinal Tract. Epidemiology and Preventive Implications.
Introduction: Alcohol is a carcinogen for human cancer. This contribution summarizes the relationships between alcohol use and gastrointestinal cancers, and implications for prevention. Methods: Comparative risk assessment and narrative literature review. Results: The following gastrointestinal cancer sites were found to be causally impacted by alcohol use: lip and oral cavity, pharynx other than nasopharynx, esophagus, colon and rectum, and liver. Globally, 368,000 deaths (304,000 men and 64,000 women) and more than 10 million disability-adjusted life years (DALYs) lost (10.1 million; 8.4 million men and 1.6 million women) in 2016 were attributable to alcohol use, making up about 10% of all deaths and DALYs lost due to these cancers, respectively. There are effective and cost-effective alcohol control policies available to reduce this burden, namely the best buys of increasing taxation, reducing availability, and banning advertisement. In addition, public knowledge about the alcohol-cancer link should be increased. Discussion: There are a number of assumptions underlying these estimates, but overall all of them seem to be conservative.


Noninvasive determination of speech dominance by single magnetic stimulation of the bilateral hand motor cortex.
Magnetic stimulation of the hand area of the motor cortex in both hemispheres was performed at rest and during reading aloud to observe modulated facilitation of hand muscle motor potentials in 6 right-handed patients, with supratentorial lesions but no motor impairment or aphasia, who had undergone the Wada test to determine speech dominance, showing that 5 were left hemisphere dominant and one was bilateral hemisphere dominant. Motor potentials were facilitated during reading aloud in only the right hand in 3 patients, all left hemisphere dominant, greater in the right hand in one, left hemisphere dominant, and greater in the left hand in one patient, bilateral hemisphere dominant. Based on these results we defined a laterality index which was consistent with the Wada test results. Magnetic stimulation may prove useful for determining cerebral dominance, as our method correlates well with the Wada test, and is safe, convenient, and inexpensive.


Biochemical analysis and quantification of hematopoietic stem cells by infrared spectroscopy.
Identification of hematopoietic stem cells (HSCs) in different stages of maturation is one of the major issues in stem cell research and bone marrow (BM) transplantation. Each stage of maturation of HSCs is characterized by a series of distinct glycoproteins present on the cell plasma membrane surface, named a cluster of differentiation (CD). Currently, complicated and expensive procedures based on CD expression are needed for identification and isolation of HSCs. This method is under dispute, since the correct markers' composition is not strictly clear, thus there is need for a better method for stem cell characterization. In the present study, Fourier transform infrared (FTIR) spectroscopy is employed as a novel optical method for identification and characterization of HSCs based on their entire biochemical features. FTIR spectral analysis of isolated mice HSCs reveals several spectral markers related to lipids, nucleic acids, and carbohydrates, which distinguish HSCs from BM cells. The unique "open" conformation of HSC DNA as identified by FTIR is exploited for HSCs quantification in the BM. The proposed method of FTIR spectroscopy for HSC identification and quantification can contribute to stem cell research and BM transplantation.


Neither linoleic acid nor arachidonic acid promote white adipose tissue inflammation in Fads2-/- mice fed low fat diets.
Dietary n-6 polyunsaturated fatty acids (PUFA) are widely perceived to promote inflammation and contribute to the development of chronic diseases. This dogma has been recently questioned due to evidence that n-6 PUFA, specifically linoleic acid (LA, 18:2n-6) and arachidonic acid (AA, 20:4n-6), do not appear to activate inflammatory signalling pathways when consumed in moderate amounts. However, delineating the independent roles of different dietary n-6 PUFA in vivo is challenging because LA is continuously converted into AA in a pathway regulated by the fatty acid desaturase 2 (Fads2) gene. The objective of this study was to investigate the independent roles of LA and AA on white adipose tissue (WAT) inflammatory signalling pathways using Fads2-/- mice. We hypothesized that dietary LA would not induce WAT inflammation, unless it was endogenously converted into AA. Male C57BL/6 wild-type (WT) and Fads2-/- mice were fed low-fat isocaloric diets containing either 7% corn oil w/w (CD, containing ~42% LA) or 7% ARASCO oil w/w (AD, containing ~27% AA) for 9 weeks. WAT inflammatory gene expression, protein levels, as well as phospholipid (PL) and triacylglycerol (TAG) fatty acid composition, were analyzed by RT-qPCR, western blots, and gas chromatography, respectively. Fads2-/- mice fed CD had high LA, but little-to-no GLA (18:3n-6), DGLA (20:3n-6), and AA in PLs and TAGs compared to their WT counterparts. In comparison, Fads2-/- and WT mice fed AD showed minimal differences in n-6 PUFA content in serum and WAT, despite having significantly more AA than CD-fed mice. No differences in gene expression for common inflammatory adipokines (e.g. Mcp-1, Ccl5, Tnfα) or key regulators of eicosanoid production (e.g. Cox-2, Alox-12, Alox-15) were detected in WAT between any of the diet and genotype groups. Furthermore, no differences in MCP-1, and total or phosphorylated STAT3 and p38 inflammatory proteins, were observed. Collectively, these results demonstrate that neither LA nor AA promote WAT inflammation when consumed as part of a low-fat diet. Therefore, the existing dogma surrounding n-6 PUFA and inflammation needs to be reconsidered.


Emotional Dysregulation and Temperament-Character Traits in Adolescents With Functional Neurological Symptom Disorder (Conversion Disorder).
The purpose of this study was to evaluate emotion dysregulation and temperament-character traits in adolescents with functional neurological symptom disorder (FNSD). Forty adolescents with FNSD and 40 healthy adolescents were evaluated by a semiconstructed diagnosis interview, Temperament and Character Inventory (TCI), Difficulties in Emotion Regulation Scale (DERS), Regulation of Emotions Questionnaire (REQ), and Children's Somatization Inventory-24 (CSI-24). The external and internal dysfunctional emotion regulation scores of REQ, all subscales of DERS, except the awareness subscale, and CSI-24 scores were significantly higher in FNSD patients compared with healthy controls. There were significant differences between the groups in terms of harm avoidance and reward dependence subscale scores of TCI. Multiple logistic regression analysis showed that the external dysfunctional emotion regulation strategy, somatization, and reward dependence are significant predictors of FNSD. Our results provide evidence that adolescents with FNSD experience emotional dysregulation and that the differential value of some temperament-character traits in the diagnosis of FNSD.


The effects of improving hospital physicians working conditions on patient care: a prospective, controlled intervention study.
Physicians, particularly in hospitals, suffer from adverse working conditions. There is a close link between physicians' psychosocial work environment and the quality of the work they deliver. Our study aimed to explore whether a participatory work-design intervention involving hospital physicians is effective in improving working conditions and quality of patient care. A prospective, controlled intervention study was conducted in two surgical and two internal departments. Participants were 57 hospital physicians and 1581 inpatients. The intervention was a structured, participatory intervention based on continuous group meetings. Physicians actively analyzed problematic working conditions, developed solutions, and initiated their implementation. Physicians' working conditions and patients' perceived quality of care were outcome criteria. These variables were assessed by standardized questionnaires. Additional data on implementation status were gathered through interviews. Over the course of ten months, several work-related problems were identified, categorized, and ten solutions were implemented. Post-intervention, physicians in the intervention departments reported substantially less conflicting demands and enhanced quality of cooperation with patients' relatives, compared to control group physicians. Moreover, positive changes in enhanced colleague support could be attributed to the intervention. Regarding patient reports of care quality of care, patient ratings of physicians organization of care improved for physicians in the intervention group. Five interviews with involved physicians confirm the plausibility of obtained results, provide information on implementation status and sustainability of the solutions, and highlight process-related factors for re-design interventions to improve hospital physicians work. This study demonstrates that participatory work design for hospital physicians is a promising intervention for improving working conditions and promoting patient quality of care.