Activation of Clostridium botulinum type E toxin purified by two different procedures.
Clostridium botulinum type E toxin was purified from culture supernates and from cell extracts by two methods. The specific activity [2 X 10(4) mouse LD50 (mg protein)-1] of the toxin purified from cell extract under slightly acidic conditions was lower than that [3 X 10(5) LD50 (mg protein)-1] of the toxin purified from culture supernate under slightly alkaline conditions. Both toxin preparations were activated by trypsin treatment, but to different extents, the degree of activation of the toxin from cell extract being about 30-fold higher than that of the toxin from culture supernate. The two toxin preparations had the same electrophoretic mobility on SDS-polyacrylamide gels and antigenic specificity as revealed by agar gel double-immunodiffusion tests. The antigenic specificity of the two toxin preparations was unaltered by trypsin treatment. In SDS-polyacrylamide gel electrophoresis, a single band of Mr 144,000 was demonstrated before trypsin treatment and two bands of Mr 100,000 and 55,000 appeared after trypsin treatment. The two toxin preparations were labelled with 125I and chymotryptic peptide maps were obtained before and after trypsin treatment. The two toxin preparations without trypsin treatment demonstrated many differences in their peptide maps, but the preparations after trypsin activation had similar peptide maps. These results indicate that the toxin obtained from culture fluid was a partially activated form, and that its molecular conformation was different from that of the toxin from cell extract. Differences in specific activity and activation ratio by trypsin treatment may be due to differences in the conformation of the toxin molecules.


Prediction of non-perfusion volume ratio for uterine fibroids treated with ultrasound-guided high-intensity focused ultrasound based on MRI radiomics combined with clinical parameters.
Prediction of non-perfusion volume ratio (NPVR) is critical in selecting patients with uterine fibroids who will potentially benefit from ultrasound-guided high-intensity focused ultrasound (HIFU) treatment, as it reduces the risk of treatment failure. The purpose of this study is to construct an optimal model for predicting NPVR based on T2-weighted magnetic resonance imaging (T2MRI) radiomics features combined with clinical parameters by machine learning. This retrospective study was conducted among 223 patients diagnosed with uterine fibroids from two centers. The patients from one center were allocated to a training cohort (n = 122) and an internal test cohort (n = 46), and the data from the other center (n = 55) was used as an external test cohort. The least absolute shrinkage and selection operator (LASSO) algorithm was employed for feature selection in the training cohort. The support vector machine (SVM) was adopted to construct a radiomics model, a clinical model, and a radiomics-clinical model for NPVR prediction, respectively. The area under the curve (AUC) and the decision curve analysis (DCA) were performed to evaluate the predictive validity and the clinical usefulness of the model, respectively. A total of 851 radiomic features were extracted from T2MRI, of which seven radiomics features were screened for NPVR prediction-related radiomics features. The radiomics-clinical model combining radiomics features and clinical parameters showed the best predictive performance in both the internal (AUC = 0.824, 95% CI 0.693-0.954) and external (AUC = 0.773, 95% CI 0.647-0.902) test cohorts, and the DCA also suggested the radiomics-clinical model had the highest net benefit. The radiomics-clinical model could be applied to the NPVR prediction of patients with uterine fibroids treated by HIFU to provide an objective and effective method for selecting potential patients who would benefit from the treatment mostly.


Compare the performance of the models in art classification.
Because large numbers of artworks are preserved in museums and galleries, much work must be done to classify these works into genres, styles and artists. Recent technological advancements have enabled an increasing number of artworks to be digitized. Thus, it is necessary to teach computers to analyze (e.g., classify and annotate) art to assist people in performing such tasks. In this study, we tested 7 different models on 3 different datasets under the same experimental setup to compare their art classification performances when either using or not using transfer learning. The models were compared based on their abilities for classifying genres, styles and artists. Comparing the result with previous work shows that the model performance can be effectively improved by optimizing the model structure, and our results achieve state-of-the-art performance in all classification tasks with three datasets. In addition, we visualized the process of style and genre classification to help us understand the difficulties that computers have when tasked with classifying art. Finally, we used the trained models described above to perform similarity searches and obtained performance improvements.


Assessment of knowledge of chronic kidney disease among non-nephrology nurses in Akure, South-West Nigeria.
Health workers require adequate knowledge of chronic kidney disease (CKD) to be able to play their role in reducing the burden of CKD. Most previous studies focused on assessing knowledge of doctors on CKD; however, nurses are also important in primary, secondary, and tertiary prevention of CKD. This study assessed the knowledge of non-nephrology nurses on CKD with the aim of identifying areas of knowledge gaps which will be targets for future educational programs. This was a cross-sectional descriptive study carried out among nurses in Akure, Southwest Nigeria during their mandatory continuing professional development program required for practicing license renewal. Knowledge of CKD was assessed using self-administered pretested questionnaires. P <0.05 was taken as significant. One-hundred nurses participated in the study with a male:female ratio of 1:3.7. The mean duration of their nursing experience was 14.5 ± 9.1 years. Only 15% had nephrology posting during their training. Six (6%) of the respondents had good knowledge of CKD, 55 (55%) had fair knowledge, and 37 (37%) had poor knowledge. Only 5% was aware of renal care policy in Nigeria. Junior and intermediate cadre nurses had better knowledge of CKD than senior cadre nurses (P = 0.004). Nurses who had nephrology posting during their training had significantly higher mean knowledge score than others (14.38 ± 2.25 vs. 12.93 ± 3.10, P = 0.036). There were significant deficiencies in the knowledge of CKD among non-nephrology nurses who participated in the study. Junior and intermediate cadre nurses and those who had nephrology postings had better knowledge of CKD.


Opportunities for improving use of evidence-based therapy in patients with type 2 diabetes and cardiovascular disease.
Evidence-based therapy that target hyperlipidemia, hypertension, smoking cessation, and weight loss have demonstrated significant benefits in reducing cardiovascular risks and related events. Although the benefit of intensively lowering blood glucose is unclear, newer antidiabetic drugs (glucagon-like peptide-1 receptor agonists and sodium-glucose cotransporter-2 inhibitors) have shown cardiovascular benefits in addition to their antihyperglycemic effect. Yet, studies suggest that recent use of evidence-based therapy and management of cardiovascular risk among individuals with type 2 diabetes (T2D) and cardiovascular disease (CVD) remains largely suboptimal. The following narrative review first identifies barriers to translating research evidence to clinical practice at the levels of provider, health system, patient, and cost. Then it synthesizes previous implementation strategies that addressed multifaceted barriers and attempted to improve care for patients with T2D and CVD. In conclusion, team-based care coordination, reminding systems in combination to pharmacist consultation and patient education, provider education compatible with clinical workflow, and coupled incentives between providers and patients appeared to be effective in reducing cardiovascular risks for patients with T2D and CVD, though the scalability and long-term clinical effect of these strategies as well as the possibility of interventions involving payers and health systems remain uncertain.


A Single-Chain Photoswitchable CRISPR-Cas9 Architecture for Light-Inducible Gene Editing and Transcription.
Optical control of CRISPR-Cas9-derived proteins would be useful for restricting gene editing or transcriptional regulation to desired times and places. Optical control of Cas9 functions has been achieved with photouncageable unnatural amino acids or by using light-induced protein interactions to reconstitute Cas9-mediated functions from two polypeptides. However, these methods have only been applied to one Cas9 species and have not been used for optical control of different perturbations at two genes. Here, we use photodissociable dimeric fluorescent protein domains to engineer single-chain photoswitchable Cas9 (ps-Cas9) proteins in which the DNA-binding cleft is occluded at baseline and opened upon illumination. This design successfully controlled different species and functional variants of Cas9, mediated transcriptional activation more robustly than previous optogenetic methods, and enabled light-induced transcription of one gene and editing of another in the same cells. Thus, a single-chain photoswitchable architecture provides a general method to control a variety of Cas9-mediated functions.


Recalcitrant atopic dermatitis due to allergy to Compositae.
Atopic dermatitis is often complicated by allergic contact dermatitis, although patch testing may reveal positive reactions of uncertain relevance. We report a case of a 35-year-old woman with recalcitrant atopic dermatitis, with a positive patch-test reaction to Compositae mix (CM). Initially, sensitization appeared to be of past relevance only, due to use of calendula. However, it turned out that she followed a self-devised diet consisting largely of food products of the Compositae family. On excluding these food products her skin condition improved quickly. This case report underscores the difficulty in determining the relevance of positive patch tests, and shows that thorough analysis of positive patch tests, by both patient and physician, may reveal unexpected or less common sources of contact allergens.


Sequence-Responsive Multifunctional Supramolecular Nanomicelles Act on the Regression of TNBC and Its Lung Metastasis via Synergic Pyroptosis-Mediated Immune Activation.
Design of effective nanodrugs to modulate the immunosuppression of tumor microenvironment is a desirable approach to boost the clinical tumor-therapeutic effect. Supramolecular nanomicelles PolyMN-TO-8, which are constructed by self-assembling supramolecular host MTX-MPEG2000, guest NPX-2S, and TO-8 through hydrophobic forces, have excellent stability and responsiveness to carboxylesterase and glutathione in turn. In vivo studies validate that PolyMN-TO-8 enable to trigger pyroptosis-mediated immunogenic cell death under laser, avoiding the occurrence of immune dysregulation simultaneously. This therapeutic mode strengthens dendritic cells' maturation and accelerates the infiltration of CD8+ T cells into tumors through moderate activation of pro-inflammatory factors with elimination of immune-escape, ultimately making the tumor inhibition rate as high as 87.44% via synergistic functions of photodynamic therapy, photothermal therapy, chemotherapy, etc. The loss of immune-escape quickens the infiltration of CD8+ T cells into lungs, and further eschews the generation of tumor nodules in it. Chemotherapy, the release of interferon-γ, and immune memory effect also strengthen the defense against metastasis. The generation of O2 catalyzed by PolyMN-TO-8 under laser is indispensable for tumor metastasis inhibition undoubtedly.


Investigator Attitudes on Equipoise and Practice Patterns in the BEST-CLI Trial.
There has been significant variability in practice patterns and equipoise regarding treatment approach for chronic limb threatening ischemia (CLTI). We aimed to assess treatment preferences of BEST-CLI investigators prior to and following the trial. An electronic 60 question survey was sent to 1180 BEST-CLI investigators in 2022 after trial conclusion and before announcement of results. Investigators' preferences were assessed across clinical scenarios for both open (OPEN) and endovascular (ENDO) revascularization strategies. Vascular surgeon surgical and endovascular preferences were compared to a 2010 survey administered to prospective investigators before trial funding. For the 2022 survey, the response rate was 20.2% and was comprised of vascular surgeons (76.3%), interventional cardiologists (11.4%) and interventional radiologists (11.6%). The majority (72.6%) were in academic practice and 39.1% were in practice for more than 20 years. During initial CLTI work-up, 65.8%, 42.6%, and 55.9% of respondents always or usually ordered an arterial duplex, computed tomography angiography, and vein mapping, respectively. The most common practice distribution between endovascular and open procedures was 70/30. Postoperatively, a majority reported performing routine duplex surveillance of vein bypass (99%), prosthetic bypass (81.9%), and endovascular interventions (86%). A minority reported always or usually using WIfI (25.8%), GLASS (8.3%), and a risk calculator (14.8%). More than half (52.9%) agreed that the statement "no bridges are burned with an ENDO first approach" was false. Intervention choice was influenced by availability of the operating room or endovascular suite, personal schedule, and personal skill set in 30.1%, 18% and 45.9% of respondents, respectively. Most respondents reported routinely using Paclitaxel coated balloons (88.1%) and stents (67.5%), however 73.3% altered practice when safety concerns were raised. Among surgeons, 17.8%, 2.9%, and 10.3% reported performing more than 10 annual alternative autogenous vein bypasses, composite vein composite vein bypasses, and bypasses to pedal targets, respectively. Among all interventionalists, 8%, 24%, and 8% reported performing more than 10 annual radial access procedures, pedal or tibial access procedures, and pedal loop revascularizations. The majority (89.1%) of respondents felt that CLTI Teams improved care, however only 23.2% had a defined team. The effectiveness of the teamwork at institutions was characterized as highly effective in 42.5%. When comparing responses by vascular surgeons to the 2010 survey, there were no changes in preferred treatment based on TASC II classification or conduit preference. In 2022, open surgery was preferred more for a popliteal occlusion. For clinical scenarios, there were no differences except decreased proportion of respondents who felt there was equipoise for major tissue loss for major tissue loss (43.8% vs. 31.2%) and increased ENDO choice for minor tissue loss (17.6% vs. 30.8%) (P<.05). There is a wide range of practice patterns among vascular specialists treating CLTI. The majority of investigators in BEST-CLI had experience in both advanced open and endovascular techniques and represent a real-world sample of technical expertise. Over the course of the decade of the BEST-CLI trial there was overall similar equipoise among vascular surgeons.


Intensive intervention for children and adolescents with autism in a community setting in Italy: a single-group longitudinal study.
Previous studies have shown favourable results with intensive behavioural treatment for children with autism: evidence has emerged that treatment can be successfully implemented in a community setting and in adolescent participants. The aim of this study was to describe the 2-year adaptive functioning outcome of children and adolescents with autism treated intensively within the context of special autism centres, as well as to evaluate family satisfaction with the activity of the centres. Sixty participants with autism (20 females and 40 males, aged between 4 and 18 years) attending the semi-residential rehabilitation centres for autism located in the Abruzzo region (Central Italy) were followed up and their adaptive functioning was evaluated both at baseline and after one and two years using the Vineland Adaptive Behaviour Scales (VABS). Parents' satisfaction with the service was evaluated using the Orbetello Satisfaction Scale for Children and Adolescent Mental Health. The increase in VABS scores was significant on several domains in the different gender and age categories. It is worth noting that male children had improved a great deal (roughly, an effect size >0.20) in the domains of communication, daily living and motor skills (effect sizes 0.34, 0.45 and 0.27 respectively) whereas in male adolescents, a notable increase in VABS scores was recorded in the domain of socialization only (effect size 0.23). On the other hand, adaptive behaviour in female children increased in the domains of socialization and motor skills (effect sizes 0.27 and 0.42 respectively) whereas in female adolescents, good results were achieved in the domains of daily living, socialization and motor skills (effect sizes 0.22, 0.26 and 0.20 respectively).The level of satisfaction of users of the service over time was found to be substantial, even when they had recently started the program. Our results support the implementation of special autism treatment community centres, based on a parent co-directed rehabilitative, intensive and early intervention. Further experimental research designed to document the effectiveness of services provided to children and adolescents with autism in the community is recommended.


Mapping social accountability actors and networks and their roles in water, sanitation and hygiene (WASH) in childcare centres within Nairobi's informal settlements: A governance diaries approach.
Despite many institutions gaining access to improved water sanitation and hygiene (WASH) services, childcare centres in informal settlements have low access and poor condition of WASH services. It is imperative to understand how existing actors and social networks operate in the WASH sector in childcare centres in Nairobi's informal settlements. To empirically map and understand how different actors within informal settlements influence the provision of adequate and quality water, sanitation and hygiene services within childcare centres in Nairobi's informal settlements. This was a qualitative study. We conducted an ethnographic study using governance diaries with 24 participants from Korogocho and Viwandani informal settlements in Nairobi, Kenya. The governance diaries approach involved conducting bi-weekly governance in-depth interviews (IDIs) with study participants for 4 months, complemented with observations, reflections, participant diaries and informal discussions. We used a framework analysis which is partly deductive, informed by the governance framework and stakeholder framework. Social accountability actors were individuals or groups involved in WASH service provision in childcare centres. The actors included both key actors (actors who are primary to meeting the day-to-day WASH service needs of children) and non-key actors (actors operating in the WASH sector but not always present for day-to-day provision in childcare centres). The key actors were unanimously identified as childcare centre owners/teachers and parents/guardians as they had a more direct role in the provision of WASH services in childcare centres. The actors had direct, possible or desired networks, with the direct networks portrayed more by the parents and childcare centre owners, whose roles included acting as a voice and responding to the WASH service needs of children as it relates to access and quality. Centre owners had more power/authority over WASH services for children in childcare centres than the parents. Key actors derived power by their discretion depending on whether a decision was beneficial to children or not. Lastly, the interest of key actors were diverse ranging from income generation, access to WASH services by children, compliance with government regulations, and promotion of child health, to the prevention of the spread of diseases. Our study highlights that parents and childcare owners play an important role in WASH service provision. While service providers and other players may be statutorily given primary responsibilities for WASH provision, and more visible in official standing, among study participants they are not seen as primary actors but secondary players with ancillary responsibilities. We conclude that WASH service provision in child care centres may be realised when key actors have a voice and work within networks to demand WASH services from desired networks including the government. We also conclude that developing more direct networks and converting desired and potential networks into direct networks in WASH service provision is critical for the success of WASH service delivery. Lastly, actors in WASH services in childcare centres may need to collaborate in identifying potential avenues for strengthening existing networks that enhance access and quality of WASH services in childcare centres.


The infrequency of "pure culture" diagnoses among the anxiety disorders.
Anxiety disorders are known to commonly coexist in individuals, both with other anxiety disorders and with mental disorders from other groupings, such as affective disorders. We questioned how frequently anxiety disorders actually occur in isolation, as "pure cultures." We examined diagnostic patterns among the 711 subjects entered into a large, multicenter study of anxiety disorders, the Harvard/ Brown Anxiety Disorders Research Program (HARP), which focused on panic, agoraphobia, generalized anxiety disorder, and social phobias as "index disorders" required for intake. We used various definitions for "pure culture." By all definitions, subjects with "pure culture" represented a minority, especially in cases of generalized anxiety disorder and social phobia, where comorbidity was virtually ubiquitous. "Pure culture" status was associated with later onset of illness and less chronicity. Future studies of anxiety disorder should aim to document the extensive comorbidity, rather than eliminate it by restrictive diagnostic exclusion criteria, lest they yield atypical or even misrepresented groups of patients. Clinicians should not stop at identifying only the "main" diagnosis but look for other, comorbid diagnoses that are often present.


Alzheimer's disease may begin in the nose and may be caused by aluminosilicates.
Genetic factors may interact with aging changes in the nasal mucociliary apparatus to increase the probability that ubiquitously occurring aluminosilicates may enter sensory neurons of the olfactory epithelium and spread transneuronally to several olfactory-related areas of the brain, thereby initiating changes that eventually result in neuronal damage typical of Alzheimer's disease. A speculative sequence of events is suggested by which neuronally-contained aluminosilicates might cleave or otherwise alter a normal cellular protein in such a manner that aggregates would arise that could interfere with cellular function and which also could act in a pseudo-infective manner, relaxing translational and transcriptional controls in the synthesis of the native protein. Some relevant experiments and potential therapies arising from the hypothesis presented are discussed.


Novel chemokine-like activities of histones in tumor metastasis.
Histones are intracellular nucleosomal components and extracellular damage-associated molecular pattern molecules that modulate chromatin remodeling, as well as the immune response. However, their extracellular roles in cell migration and invasion remain undefined. Here, we demonstrate that histones are novel regulators of tumor metastasis with chemokine-like activities. Indeed, exogenous histones promote both hepatocellular carcinoma (HCC) cell migration and invasion through toll-like receptor (TLR)4, but not TLR2 or the receptor for advanced glycosylation end product. TLR4-mediated activation of nuclear factor-κB (NF-κB) by extracellular signal-regulated kinase (ERK) is required for histone-induced chemokine (e.g., C-C motif ligand 9/10) production. Pharmacological and genetic inhibition of TLR4-ERK-NF-κB signaling impairs histone-induced chemokine production and HCC cell migration. Additionally, TLR4 depletion (by using TLR4-/- mice and TLR4-shRNA) or inhibition of histone release/activity (by administration of heparin and H3 neutralizing antibody) attenuates lung metastasis of HCC cells injected via the tail vein of mice. Thus, histones promote tumor metastasis of HCC cells through the TLR4-NF-κB pathway and represent novel targets for treating patients with HCC.


Patient Satisfaction With Maternity Waiting Homes in Liberia: A Case Study During the Ebola Outbreak.
Liberia in West Africa has one of the highest maternal mortality ratios in the world (990/100,000 live births). Many women in Liberia live in rural, remote villages with little access to safe maternity services. The World Health Organization has identified maternity waiting homes (MWHs) as one strategy to minimize the barrier of distance in accessing a skilled birth attendant. However, limited data exist on satisfaction with MWHs or maternal health care in Liberia. This mixed-methods case study examines women's satisfaction with their stay at a MWH and compares utilization rates before and during the Ebola outbreak. From 2012 to 2014, 650 women who stayed at one of 6 MWHs in rural Liberia during the perinatal or postnatal period were surveyed. Additionally, 60 semi-structured interviews were conducted with traditional providers, skilled birth attendants, and women utilizing the MWHs. Quantitative analyses assessed satisfaction rates before and during the Ebola outbreak. Content analysis of semi-structured interviews supplemented the quantitative data and provided a lens into the elements of satisfaction with the MWHs. The majority of women who utilized the MWHs stated they would suggest the MWH to a friend or relative who was pregnant (99.5%), and nearly all would utilize the home again (98.8%). Although satisfaction with the MWHs significantly decreased during the Ebola outbreak (P < .001), participants were satisfied overall with the MWHs. Content analysis identified areas of satisfaction that encompassed the themes of restful and supportive environment as well as areas for improvement such as lacking necessary resources and loneliness. This case study demonstrated that women using MWHs in Bong County, Liberia are generally satisfied with their experience and plan to use an MWH again during future pregnancies to access a skilled birth attendant for birth. Women are also willing to encourage family and friends to use MWHs.


Comparison and characterization of nuclear isolation procedures as applied to chick oviduct.
A number of methods for the preparation of chick oviduct nuclei have been compared. Nuclei have been isolated in hypertonic sucrose and citric acid and the product has been characterized with respect to cleanliness, ultrastructure, RNA polymerase activity, RNA integrity, and chromatin composition. The study demonstrates that the choice of oviduct nuclear isolation procedure will depend markedly on the purpose for which the nuclei are required. Thus, nuclei prepared entirely in high-molarity sucrose retain the highest levels of RNA polymerase. Those prepared rapidly in the presence of citric acid retain nuclear RNA in an essentially undegraded state. Finally, a bulk preparation is described which, because of its adaptability and high yield of morphologically intact nuclei using large amounts of tissue, is ideal for use in preparing chromatin. Conditions are described by which isolated nuclei can be stored for up to 6 months and retain their morphology, chemical characteristics, and RNA polymerase activity.


Analysis of sugarcane herbicides in marine turtle nesting areas and assessment of risk using in vitro toxicity assays.
Agricultural processes are associated with many different herbicides that can contaminate surrounding environments. In Queensland, Australia, herbicides applied to agricultural crops may pose a threat to valuable coastal habitats including nesting beaches for threatened loggerhead turtles (Caretta caretta). This study 1) measured concentrations of herbicides in the beach sand of Mon Repos, an important marine turtle nesting beach in Australia that is adjacent to significant sugarcane crops, and 2) investigated the toxicity of these herbicides to marine turtles using a cell-based assay. Samples of sand from turtle nest depth and water from surrounding agricultural drains and wetlands were collected during the wet season when herbicide runoff was expected to be the greatest and turtles were nesting. Samples were extracted using solid phase extraction and extracts were analysed using chemical analysis targeting herbicides, as well as bioanalytical techniques (IPAM-assay and loggerhead turtle skin cell cytotoxicity assay). Twenty herbicides were detected in areas between sugarcane crops and the nesting beach, seven of which were also detected in the sand extracts. Herbicides present in the nearby wetland were also detected in the beach sand, indicating potential contamination of the nesting beach via the river outlet as well as ground water. Although herbicides were detected in nesting sand, bioassays using loggerhead turtle skin cells indicated a low risk of acute toxicity at measured environmental concentrations. Further research should investigate potentially more subtle effects, such as endocrine disruption and mixture effects, to better assess the threat that herbicides pose to this population of marine turtles.


Breaking the 200 nm limit for routine flow linear dichroism measurements using UV synchrotron radiation.
The first synchrotron radiation flow linear dichroism spectra are reported. High-quality spectral data can be collected from 450 nm down to 180 nm in contrast to the practical cutoff of approximately 200 nm on benchtop instruments. State-of-the-art microvolume capillary Couette flow linear dichroism was successfully ported to a synchrotron radiation source. The sample volume required is < 50 microL. A characterization of the synchrotron radiation linear dichroism with known DNA and DNA-ligand systems is presented and the viability of the setup confirmed. Typically, wavelengths down to 180 nm are now routinely accessible with a high signal/noise ratio with little limitation from the sample concentration. The 180 nm cutoff is due to the quartz of the Couette cell rather than the beamline itself. We show the application of the simultaneous determination of the sample absorption spectrum to calculate the reduced linear dichroism signal. Spectra for calf thymus DNA, DNA/ethidium bromide, and DNA/4',6-diamidino-2-phenylindole systems illustrate the quality of data that can be obtained.


Youths' understandings of cigarette advertisements.
This study addresses two questions: (1) when youths are exposed to advertisements for cigarettes, do they primarily see advertisements for brands or products, and (2) is there a relationship between youths' understandings of cigarette advertisements and their susceptibility to smoking? A sample of 271 participants ranging in age from 7 to 12 viewed a series of print advertisements that included cigarette and non-tobacco-related ads. While viewing each ad, participants were asked to indicate what they thought the advertisement was trying to sell. Responses were coded into one of three categories reflecting important differences in participants' comprehension of each advertisement - no understanding, product category understanding, or brand understanding. Results show that youths typically understand the type of product an advertisement is promoting; however, the levels of brand understanding observed for cigarette advertisements were low in an absolute sense, and significantly lower than brand understanding of non-tobacco-related advertisements. Results also show that understanding cigarette ads as promoting specific brands of cigarettes is positively related to susceptibility to smoking. Taken together, these findings provide a glimpse of the psychological mechanisms that may underlie the well established link between exposure to cigarette advertising and youth smoking.


NOX1 inhibition attenuates the development of a pro-tumorigenic environment in experimental hepatocellular carcinoma.
The poor prognosis of advanced HCC and limited efficacy of current systemic treatments emphasize the need for new or combined targeted therapies. The development of HCC is a multistage process in which liver injury appears in a complex microenvironment associated with oxidative stress. NOX enzymes are the main source of ROS during hepatocarcinogenesis and NOX1 in particular has shown correlation with poor prognosis of HCC patients. This study evaluates the effect of pharmacological NOX1 inhibition on the development and progression of HCC and its effect on the tumor microenvironment. The in vitro cytotoxic effects of the NOX1 inhibitor GKT771 (Genkyotex) on human Huh7 and Hep3B and murine Hepa1-6 HCC cell lines, the human THP1 monocyte cell line and mouse macrophages were evaluated via MTT, LDH activity and CaspGlo® assays. In order to induce in vivo HCC, male SV129 wild-type mice received weekly IP injections of diethylnitrosamine (DEN) (35 mg/kg) for 20-25 weeks. Mice were treated with vehicle or GKT771 (30 mg/kg) via oral gavage, daily or twice daily, in preventive and therapeutic studies. The liver damage was evaluated for inflammation, angiogenesis, fibrosis and HCC development via histology, RT-qPCR, multiplex analyses and ROS levels. A concentration-dependent reduction in cellular activity of the human HCC cell lines without cytotoxicity was observed. GKT771 treatment reduced LPS-induced pro-inflammatory bone-marrow derived macrophage polarization. DEN injections resulted in 100 % tumor formation and the induction of HCC markers which could be reduced by twice daily dosing of GKT771 at early onset of advanced HCC. DEN-induced HCC resulted in an upregulation of pro-inflammatory, angiogenic and fibrotic markers which was less pronounced in GKT771 treated mice in all treatment regimens. In line, liver fibrosis was induced in HCC mice and this to a lesser extend upon GKT771 treatment. NOX1 inhibition showed to be safe and well tolerated and was able to attenuate the induction of a pro-inflammatory, angiogenic and pro-fibrotic microenvironment suggesting that this might be a promising adjuvant therapeutic strategy in the treatment of advanced HCC.


Inflammation-induced changes in expression and glycosylation of genetic variants of alpha 1-acid glycoprotein. Studies with human sera, primary cultures of human hepatocytes and transgenic mice.
The relative occurrence of genetic variants of human alpha 1-acid glycoprotein (AGP) in relation to changes in glycosylation was studied in sera of patients with burn injury, media of cytokine-treated primary cultures of human hepatocytes and Hep 3B cells, and sera of transgenic mice expressing the human AGP-A gene. It is concluded (i) that the glycosylation of AGP was not dependent on its genetic expression and (ii) that both the variants determined by the AGP-A gene as well as by the AGP-B/B' genes are increased after inflammation or treatment with interleukins 1 and 6.


Weight loss strategies used by African American women: possible implications for tailored messages.
African American women have the highest rates of obesity in the USA. They are less likely to participate in weight loss programmes and are less successful in their weight loss attempts. The present study aimed to identify weight loss strategies used by African American women and to determine whether those strategies varied by weight status. The study also examined the emotional issues that lead to compulsive overeating and identified the types of information that African American women need to help manage their weight. A self-administered survey was completed by 413 adult African American women in 2009. Participants were weighed and measured by a member of the research team. Based on body mass index classification, 43% of women were obese, 25% were overweight, 30% were normal weight and 2% were underweight. Sixty percent had tried to lose weight in the past 12 months. Weight loss practices included: cutting back on fried foods (53%), cutting back on sweets (51%) and increasing physical activity, skipping meals (22%), fasting (17%), and using diet pills (7%). Obese women were significantly more likely to fast and use diet pills than overweight women (P < 0.001). Obese women also were significantly more likely to say they wanted information on how to choose a weight loss programme (P < 0.001), manage stress (P < 0.001) and increase self-esteem (P < 0.001). Our findings suggest that weight loss messages and programmes may need to be specifically developed for obese women compared to overweight and normal weight women who just need to 'lose a few pounds'.


Diverse evolutionary pathways challenge the use of collateral sensitivity as a strategy to suppress resistance.
Drug resistance remains a major obstacle to malaria control and eradication efforts, necessitating the development of novel therapeutic strategies to treat this disease. Drug combinations based on collateral sensitivity, wherein resistance to one drug causes increased sensitivity to the partner drug, have been proposed as an evolutionary strategy to suppress the emergence of resistance in pathogen populations. In this study, we explore collateral sensitivity between compounds targeting the Plasmodium dihydroorotate dehydrogenase (DHODH). We profiled the cross-resistance and collateral sensitivity phenotypes of several DHODH mutant lines to a diverse panel of DHODH inhibitors. We focus on one compound, TCMDC-125334, which was active against all mutant lines tested, including the DHODH C276Y line, which arose in selections with the clinical candidate DSM265. In six selections with TCMDC-125334, the most common mechanism of resistance to this compound was copy number variation of the dhodh locus, although we did identify one mutation, DHODH I263S, which conferred resistance to TCMDC-125334 but not DSM265. We found that selection of the DHODH C276Y mutant with TCMDC-125334 yielded additional genetic changes in the dhodh locus. These double mutant parasites exhibited decreased sensitivity to TCMDC-125334 and were highly resistant to DSM265. Finally, we tested whether collateral sensitivity could be exploited to suppress the emergence of resistance in the context of combination treatment by exposing wildtype parasites to both DSM265 and TCMDC-125334 simultaneously. This selected for parasites with a DHODH V532A mutation which were cross-resistant to both compounds and were as fit as the wildtype parent in vitro. The emergence of these cross-resistant, evolutionarily fit parasites highlights the mutational flexibility of the DHODH enzyme.


Fc receptor endocytosis is controlled by a cytoplasmic domain determinant that actively prevents coated pit localization.
Macrophages and B-lymphocytes express two major isoforms of Fc receptor (FcRII-B2 and FcRII-B1) that exhibit distinct capacities for endocytosis. This difference in function reflects the presence of an in-frame insertion of 47 amino acids in the cytoplasmic domain of the lymphocyte isoform (FcRII-B1) due to alternative mRNA splicing. By expressing wild type and mutant FcRII cDNAs in fibroblasts, we have now examined the mechanism by which the insertion acts to prevent coated pit localization and endocytosis. We first identified the region of the FcRII-B2 cytoplasmic domain that is required for rapid internalization. Using a biochemical assay for endocytosis and an immuno-EM assay to determine coated pit localization directly, we found that the distal half of the cytoplasmic domain, particularly a region including residues 18-31, as needed for coated pit-mediated endocytosis. Elimination of the tyrosine residues at position 26 and 43, separately or together, had little effect on coated pit localization and a partial effect on endocytosis of ligand. Since the FcRII-B1 insertion occurs in the membrane-proximal region of the cytoplasmic domain (residue 6) not required for internalization, it is unlikely to act by physically disrupting the coated pit localization determinant. In fact, the insertion was found to prevent endocytosis irrespective of its position in the cytoplasmic tail and appeared to selectively exclude the receptor from coated regions. Moreover, receptors bearing the insertion exhibited a temperature- and ligand-dependent association with a detergent-insoluble fraction and with actin filaments, perhaps in part explaining the inability of FcRII-B1 to enter coated pits.


Educational Data Mining by Optimally Fusing Shallow and Deep Features.
In order to meet the requirement of designing the educational curriculum and extract the relevant data toward students, we in this paper utilize the deep learning architecture to study the personalized understanding and educational feature extraction algorithm of the high school and college students. Aiming at the insufficiency of deep collaborative filtering recommendation algorithm that can only discover the paths that have attracted students' attention, the requirements of intelligent course are proposed by us. First, the word vector permutation discourse poke model is extracted to produce audible vectors of message data, a well-designed learning algorithm is proposed subsequently to extract and fuse multiple heterogeneous educational data. Then, the SVD algorithm program is used to reduce the range of feature vectors. And thus, the model is completely functional and we identify unified channels and users. The method of combining hand-extracted low-level features with high-level human gesture features (image visual gate features and image deep features) automatically extracted by our adopted deep network is used to identify students' various emotions. The conclusive product is a flat semantic description that effectively detects copying and objection. Experiments are performed on a generic dataset, and we use the old-fashioned manual birth method and two well-known data sets VGG16 and fine-tune AlexNet.


Integrative Analysis of Multi-Genomic Data for Kidney Renal Cell Carcinoma.
Accounting for nine out of ten kidney cancers, kidney renal cell carcinoma (KIRC) is by far the most common type of kidney cancer. In view of limited and ineffective available therapies, understanding the genetic basis of disease becomes important for better diagnosis and treatment. The present studies are based on a single type of genomic data. These studies do not consider interactions between genomic data types and their underlying biological relationships in the disease. However, the current availability of multiple genomic data and the possibility of combining it have facilitated a better understanding of the cancer's characterization. But high dimensionality and the existence of complex interactions (within and between genomic data types) are the two main challenges of integrative methods to analyze cancer effectively. In this paper, we propose a method to build an integrative model based on Bayesian model averaging procedure for improved prediction of clinical outcome in cancer survival. The proposed method initially uses dimensionality reduction techniques to generate low-dimensional latent features for the predictive models and then incorporates interactions between them. It defines the latent features using principal components and their sparse version. It compares the predictive performance of models based on these two latent features on real data. These models also validate several ccRCC-specific cancer biomarkers previously reported in the literature. Applied on kidney renal cell carcinoma (KIRC) dataset of The Cancer Genome Atlas (TCGA), the method achieves better prediction with sparse principal components model by including latent feature interactions as compared to without including them.


The effects of neural synchronization and peripheral compression on the acoustic-reflex threshold.
This study investigates the acoustic reflex threshold (ART) dependency on stimulus phase utilizing low-level reflex audiometry [Neumann et al., Audiol. Neuro-Otol. 1, 359-369 (1996)]. The goal is to obtain optimal broadband stimuli for elicitation of the acoustic reflex and to obtain objective determinations of cochlear hearing loss. Three types of tone complexes with different phase characteristics were investigated: A stimulus that compensates for basilar-membrane dispersion, thus causing a large overall neural synchrony (basilar-membrane tone complex-BMTC), the temporally inversed stimulus (iBMTC), and random-phase tone complexes (rTC). The ARTs were measured in eight normal-hearing and six hearing-impaired subjects. Five different conditions of peak amplitude and stimulus repetition rate were used for each stimulus type. The results of the present study suggest that the ART is influenced by at least two different factors: (a) the degree of synchrony of neural activity across frequency, and (b) the fast-acting compression mechanism in the cochlea that is reduced in the case of a sensorineural hearing loss. The results allow a clear distinction of the two subjects groups based on the different ART for the utilized types and conditions of the stimuli. These differences might be useful for objective recruitment detection in clinical diagnostics.


[Lung cancer screening and endoscopic approach of the pulmonary nodule].
Lung cancer screening corresponds to any investigation carried out to detect the presence of this type of cancer before the appearance of any symptomatology. According to the results of a large multicenter study in the US (NLST), American scientific societies are currently recommending the use of the CT Thoracic Scanner with a low dose of irradiation once a year for lung cancer screening in high-risk populations. European expert groups, for the most part, reserve their recommendations after publication of the results of a major European study (NELSON). As a corollary of screening campaigns, the management of lung nodules is likely to be more and more frequent in the targeted populations. This can be performed in different ways ranging from the less invasive, the CT Scanner follow-up, to the more invasive, the surgical resection. The endoscopic approach of the pulmonary nodule, whose results were so far limited, is actually regaining interest due to the contribution of new technologies in bronchial endoscopy.


Agrobacterium strains and strain improvement: Present and outlook.
Almost 40 years ago the first transgenic plant was generated through Agrobacterium tumefaciens-mediated transformation, which, until now, remains the method of choice for gene delivery into plants. Ever since, optimized Agrobacterium strains have been developed with additional (genetic) modifications that were mostly aimed at enhancing the transformation efficiency, although an optimized strain also exists that reduces unwanted plasmid recombination. As a result, a collection of very useful strains has been created to transform a wide variety of plant species, but has also led to a confusing Agrobacterium strain nomenclature. The latter is often misleading for choosing the best-suited strain for one's transformation purposes. To overcome this issue, we provide a complete overview of the strain classification. We also indicate different strain modifications and their purposes, as well as the obtained results with regard to the transformation process sensu largo. Furthermore, we propose additional improvements of the Agrobacterium-mediated transformation process and consider several worthwhile modifications, for instance, by circumventing a defense response in planta. In this regard, we will discuss pattern-triggered immunity, pathogen-associated molecular pattern detection, hormone homeostasis and signaling, and reactive oxygen species in relationship to Agrobacterium transformation. We will also explore alterations that increase agrobacterial transformation efficiency, reduce plasmid recombination, and improve biocontainment. Finally, we recommend the use of a modular system to best utilize the available knowledge for successful plant transformation.


45,X/46,X,psu dic(Y) gonadal dysgenesis: influence of the two cell lines on the clinical phenotype, including gonadal histology.
A child born with ambiguous genitalia (Prader III) was found to have a 45,X[92.2%]/46,X,psu dic(Y)(p12)[7.8%] karyotype in peripheral blood lymphocytes. The testosterone level was consistent with that of a normal male; however, gonadotropins were elevated. Ultrasound and endoscopy of the urogenital sinus revealed well-developed Müllerian structures. At 3.5 months, the child was operated for right-sided incarcerated hernia, and the gonad situated at the inguinal region was biopsied and classified as primitive testis. Based on the presence of Müllerian structures, anatomy of external genitalia and wish of the parents, the child was assigned female gender. She underwent removal of the left gonad at 4 months during another acute surgery; histology was similar to the right gonad. The rest of the right gonad was removed at 16 months, and feminizing genitoplasty took place at 3 years. The right and left gonad contained 28 and 22% of cells with a Y chromosome, respectively. During further histological examination, dysgenetic features of the gonads were discovered. Some germ cells displayed abnormal development based on the specific expression of immunohistochemical markers (OCT3/4, TSPY, KITLG), indicating a possible risk for future malignant germ cell tumor development. Contribution of the 45,X cell line to the phenotype was also observed: the patient developed celiac disease, and her growth pattern resembled that of Turner syndrome responding to growth hormone treatment.


Grafts of brain-derived neurotrophic factor and neurotrophin 3-transduced primate Schwann cells lead to functional recovery of the demyelinated mouse spinal cord.
Experimental studies provided overwhelming proof that transplants of myelin-forming cells achieve efficient remyelination in the CNS. Among cellular candidates, Schwann cells can be used for autologous transplantation to ensure robust remyelination of lesions and to deliver therapeutic factors in the CNS. In the present study, macaque Schwann cells expressing green fluorescent protein (GFP) were infected with human immunodeficiency virus-derived vectors overexpressing brain-derived neurotrophic factor (BDNF) or Neurotrophin 3 (NT-3), two neurotrophins that also modulate glial cell biology. The ability of transgenic Schwann cells to secrete growth factors was assessed by ELISA and showed 35- and 62-fold increases in BDNF and NT-3, respectively, in transduced macaque Schwann cell supernatants. Conditioned media of BDNF- and NT-3-transduced Schwann cells reduced Schwann cell proliferation and favored their differentiation in vitro. Transgenic cells were grafted in demyelinated spinal cords of adult nude mice. Two behavioral assays showed that NT-3- and BDNF-transduced Schwann cells promoted faster and stronger functional recovery than GFP-transduced Schwann cells. Morphological analysis indicated that functional recovery correlated with enhanced proliferation and differentiation of resident oligodendrocyte progenitors and enhanced oligodendrocyte and Schwann cell differentiation. Moreover, NT-3-transduced Schwann cells provided neuroprotection and reduced astrogliosis. These results underline the potential therapeutic benefit of combining neuroprotection and activation of myelin-forming cells to restore altered functions in demyelinating diseases of the CNS.


Factors Associated With Prolonged Viral RNA Shedding in Patients with Coronavirus Disease 2019 (COVID-19).
An outbreak of coronavirus disease 2019 (COVID-19) is becoming a public health emergency. Data are limited on the duration and host factors related to viral shedding. In this retrospective study, risk factors associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA shedding were evaluated in a cohort of 113 symptomatic patients from 2 hospitals outside Wuhan. The median (interquartile range) duration of SARS-CoV-2 RNA detection was 17 (13-22) days as measured from illness onset. When comparing patients with early (<15 days) and late (≥15 days after illness onset) viral RNA clearance, prolonged SARS-CoV-2 RNA shedding was associated with male sex (P = .009), old age (P = .033), concomitant hypertension (P = .009), delayed admission to hospital after illness onset (P = .001), severe illness at admission (P = .049), invasive mechanical ventilation (P = .006), and corticosteroid treatment (P = .025). Patients with longer SARS-CoV-2 RNA shedding duration had slower recovery of body temperature (P < .001) and focal absorption on radiograph images (P < .001) than patients with early SARS-CoV-2 RNA clearance. Male sex (OR, 3.24; 95% CI, 1.31-8.02), delayed hospital admission (OR, 1.30; 95% CI, 1.10-1.54), and invasive mechanical ventilation (OR, 9.88; 95% CI, 1.11-88.02) were independent risk factors for prolonged SARS-CoV-2 RNA shedding. Male sex, delayed admission to hospital after illness onset, and invasive mechanical ventilation were associated with prolonged SARS-CoV-2 RNA shedding. Hospital admission and general treatments should be started as soon as possible in symptomatic COVID-19 patients, especially male patients.


[Rabies-free status of Switzerland following 30 years of rabies in foxes].
The European fox rabies epizootic starting in 1939 at the eastern border of Poland reached Switzerland on March 3, 1967. Rabies spread over large parts of the country until 1977, the year it caused three human deaths. In 1978 the first field trial world-wide for the oral immunization of foxes against rabies was conducted in Switzerland. Initially, the expansion of the vaccination area led to a rapid reduction in rabies cases. However, the 1990s were characterized by a recrudescence of rabies in spite of regular oral immunization of foxes. The last endemic case of rabies was diagnosed in 1996 after an adaptation of the vaccination strategy. A total of 17,109 rabies cases, of which 73% in foxes and 14% in domestic animals were diagnosed, leading to an estimated number of some 25,000 postexposure treatments in humans. To eliminate rabies, a total of 2.8 million baits containing a modified live virus were distributed--mostly by hand--in the field.


TIMP-4 is regulated by vascular injury in rats.
The role of basement membrane-degrading matrix metalloproteinases (MMPs) in enabling vascular smooth muscle cell migration after vascular injury has been established in several animal models. In contrast, the role of their native inhibitors, the tissue inhibitors of matrix metalloproteinases (TIMPs), has remained unproven despite frequent coregulation of MMPs and TIMPs in other disease states. We have investigated the time course of expression and localization of TIMP-4 in rat carotid arteries 6 hours, 24 hours, 3 days, 7 days, and 14 days after balloon injury by in situ hybridization, immunohistochemistry, and Western blot analysis. TIMP-4 protein was present in the adventitia of injured carotid arteries from 24 hours after injury. At 7 and 14 days after injury, widespread immunostaining for TIMP-4 was observed throughout the neointima, media, and adventitia of injured arteries. Western blot analysis confirmed the quantitative increase in TIMP-4 protein at 7 and 14 days. In situ hybridization detected increased expression of TIMP-4 as early as 24 hours after injury and a marked induction in neointimal cells 7 days after injury. We then studied the effect of TIMP-4 protein on the migration of smooth muscle cells through a matrix-coated membrane in vitro and demonstrated a 53% reduction in invasion of rat vascular smooth muscle cells. These data and the temporal relationship between the upregulation of TIMP-4, its accumulation, and the onset of collagen deposition suggest an important role for TIMP-4 in the proteolytic balance of the vasculature controlling both smooth muscle migration and collagen accumulation in the injured arterial wall.


Biochemical and functional changes of rat liver spheroids during spheroid formation and maintenance in culture: I. morphological maturation and kinetic changes of energy metabolism, albumin synthesis, and activities of some enzymes.
In the process of isolated single liver cells coming together to form three-dimensional spheroids, cells undergo dramatic environmental changes. How liver cells respond to these changes has not been well studied before. This study characterized the functional and biochemical changes during liver spheroid formation and maintenance. Spheroids were prepared in 6-well plates from freshly isolated liver cells from male Sprague rats by a gyrotatory-mediated method. Morphological formation, and functional and biochemical parameters of liver spheroids were evaluated over a period of 21 days in culture. Liver spheroid formation was divided into two stages, immature (1-5 days) and mature (>5 days), according to their size and shape, and changes in their functionality. Galactose and pyruvate consumption was maintained at a relatively stable level throughout the period of observation. However, glucose secretion and cellular GPT and GOT activities were higher in immature spheroids, decreased upto day 5 and remained stable thereafter. Cellular gamma-glutamyltransferase (gamma-GT) and lactate dehydrogenase (LDH) activities were initially undetectable or low and increased as spheroids matured. Albumin secretion decreased rapidly within the first 2 days and increased as spheroids matured. It is concluded that cells undergo functional and biochemical changes during spheroid formation following isolation of liver cells from intact tissue. Functionality and biochemical properties recovered and were maintained in mature spheroids. A relatively stable period (6-15 days) of functionality in mature spheroids was identified and is recommended for applications of the model.


The transmission efficiency of tomato yellow leaf curl virus by the whitefly Bemisia tabaci is correlated with the presence of a specific symbiotic bacterium species.
Tomato yellow leaf curl virus (TYLCV) (Geminiviridae: Begomovirus) is exclusively vectored by the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). TYLCV transmission depends upon a 63-kDa GroEL protein produced by the vector's endosymbiotic bacteria. B. tabaci is a species complex comprising several genetically distinct biotypes that show different secondary-symbiont fauna. In Israel, the B biotype harbors Hamiltonella, and the Q biotype harbors Wolbachia and Arsenophonus. Both biotypes harbor Rickettsia and Portiera (the obligatory primary symbionts). The aim of this study was to determine which B. tabaci symbionts are involved in TYLCV transmission using B. tabaci populations collected in Israel. Virus transmission assays by B. tabaci showed that the B biotype efficiently transmits the virus, while the Q biotype scarcely transmits it. Yeast two-hybrid and protein pulldown assays showed that while the GroEL protein produced by Hamiltonella interacts with TYLCV coat protein, GroEL produced by Rickettsia and Portiera does not. To assess the role of Wolbachia and Arsenophonus GroEL proteins (GroELs), we used an immune capture PCR (IC-PCR) assay, employing in vivo- and in vitro-synthesized GroEL proteins from all symbionts and whitefly artificial feeding through membranes. Interaction between GroEL and TYLCV was found to occur in the B biotype, but not in the Q biotype. This assay further showed that release of virions protected by GroEL occurs adjacent to the primary salivary glands. Taken together, the GroEL protein produced by Hamiltonella (present in the B biotype, but absent in the Q biotype) facilitates TYLCV transmission. The other symbionts from both biotypes do not seem to be involved in transmission of this virus.


X-ray Photographs of a Solar Active Region with a Multilayer Telescope at Normal Incidence.
An astronomical photograph was obtained with a multilayer x-ray telescope. A 4-centimeter tungsten-carbon multilayer mirror was flown as part of an experimental solar rocket payload, and successful images were taken of the sun at normal incidence at a wavelength of 44 angstroms. Coronal Si-XII emission from an active region was recorded on film; as expected, the structure is very similar to that observed at O-VIII wavelengths by the Solar Maximum Mission flat crystal spectrometer at the same time. The small, simple optical system used in this experiment appears to have achieved a resolution of 5 to 10 arc seconds.


Exaggerated peripheral responses to catecholamines contributes to stress-induced hyperglycemia in the ob/ob mouse.
The present study investigated the contribution of altered sympathetic reactivity to the stress-induced hyperglycemia observed in the c57BL/6J (ob/ob) mouse, an animal model of type II diabetes. Blood glucose and insulin responses to sympathetic agonist and antagonist administration were evaluated in ob/ob mice and their nondiabetic, lean (ob/?) littermates. In addition, the ability of nutritional status to modify these responses was determined. These studies demonstrated that epinephrine administration to ob/ob mice caused an exaggerated increase in blood glucose and decrease in plasma insulin in ob/ob mice relative to lean littermates. The dose response curve for epinephrine-induced increases in blood glucose were shifted to the left, and the duration of the blood glucose and plasma insulin responses was longer. Differences between ob/ob mice and their nondiabetic littermates were greater when animals were tested in the fasted state. In addition, administration of the alpha adrenergic antagonist phentolamine caused a larger increase in plasma insulin in ob/ob mice than was observed in lean littermates. These results suggest that altered peripheral responses to sympathetic stimuli contribute to stress-induced hyperglycemia in ob/ob mice, and raise the possibility that altered sympathetic function is an etiologic factor in development of diabetes in these animals.


Reversal of alcohol-induced learning deficits in the young adult in a model of fetal alcohol syndrome.
To evaluate whether treatment with neuroprotective peptides to young adult mice prenatally exposed to alcohol reverses alcohol-induced learning deficits in a mouse model of fetal alcohol syndrome, whether the mechanism involves the N-methyl-d-aspartate (NMDA) and gamma-aminobutyric acid type A (GABAA) receptors, and whether it is related to glial cells. C57Bl6/J mice were treated with alcohol (0.03 ml/g) or placebo on gestational day 8. On day 40, male mice exposed to alcohol in utero were treated daily for 10 days with D-NAPVSIPQ and D-SALLRSIPA (n=20) or placebo (n=13); and control offspring were treated with placebo (n=46), with the treatment blinded. Learning evaluation began after 3 days using the Morris watermaze and the T-maze. The hippocampus, cortex, and cerebellum were isolated. Expression of NR2A, NR2B, GABAAbeta3, GABAAalpha5, vasoactive intestinal peptide (VIP), activity-dependent neuroprotective protein, and glial fibrillary acidic protein was measured using calibrator-normalized relative real-time polymerase chain reaction. Statistical analysis included analysis of variance and Fisher's protected least significant difference. Treatment with D-NAPVSIPQ and D-SALLRSIPA reversed the alcohol-induced learning deficit in both learning tests as well as the NR2A and NR2B down-regulation in the hippocampus and the up-regulation of NR2A in the cortex and NR2B in the cortex and cerebellum (all P<.05). No significant differences were found in GABAA expression. Moreover, the peptides changed activity-dependent neuroprotective protein expression in the cortex (P=.016) but not the down-regulation of VIP (P=.883), probably because the peptides are downstream from VIP. Alcohol-induced learning deficit was reversed and expression of NR2A and NR2B was restored in the hippocampus and cortex of young adult mice treated with D-NAPVSIPQ and D-SALLRSIPA. Given the role of NMDA receptors in learning, this may explain in part the mechanism of prevention of alcohol-induced learning deficits by D-NAPVSIPQ and D-SALLRSIPA.


Does surgical treatment of mandibular condyle head (diacapitular) fractures provide better outcomes than closed treatment? - a systematic review and meta-analysis.
The choices for managing a condylar head fracture (CHF) of the mandible are either open surgical or closed functional treatments (CFT) and the decision depends on various factors. The purpose of this systematic review was to ascertain from the available literature whether the open method or CFT yields better outcomes in managing CHF. We have systematically reviewed published articles according to the PRISMA statement. The search was conducted in PubMed, Google Scholar, Semantic Scholar, and the Cochrane Library database for comparative studies about both open and closed treatments from inception until April 2023. The outcomes of interest were mouth opening (MO), protrusion, laterotrusion, postoperative pain, and malocclusion. Eight studies met the inclusion criteria. The review comprised of 326 cases, among which 177 were managed by open methods and 149 were treated by CFT. The incidence of postoperative malocclusion and pain were significantly less in the open group. MO was better in the open treatment group although this was not statistically significant. Protrusion and laterotrusion occurred slightly more in CFT, although these were also statistically not significant. Overall, meta-analysis favoured open methods of managing CHF. Although enough evidence exists for the use of open methods for selected condylar head fractures, CFT still demonstrated favourable outcomes in undisplaced fractures. The selection of a particular treatment method should be individualised on the basis of each particular case considering the risk/benefits. Further high quality randomised trials are needed to establish a therapeutic guideline.


Estimation of migration rates from marker-based parentage analysis.
Coupled with rapid developments of efficient genetic markers, powerful population genetic methods were proposed to estimate migration rates (m) in natural populations in much broader spatial and temporal scales than the traditional mark-release-recapture (MRR) methods. Highly polymorphic (e.g. microsatellites) and genomic-wide (e.g. SNPs) markers provide sufficient information to assign individuals to their populations or parents of origin and thereby to estimate directly m in a way similar to MRR. Such direct estimates of current migration rates are particularly useful in understanding the ecology and microevolution of wild populations and in managing the populations in the future. In this study, I proposed and implemented, in the software MigEst, a likelihood method to use marker-based parentage assignments in jointly estimating m and candidate parent sampling proportions (x) in a subset of populations, investigated its power and accuracy using data simulated in various scenarios of population properties (e.g. the actual m, number, size and differentiation of populations) and sampling properties (e.g. the numbers of sampled parent candidates, offspring and markers), compared it with the population assignment approach implemented in the software BayesAss and demonstrated its usefulness by analysing a microsatellite data set from three natural populations of Brazilian bats. Simulations showed that MigEst provides unbiased and accurate estimates of m and performs better than BayesAss except when populations are highly differentiated with very small and ecologically insignificant migration rates. A valuable property of MigEst is that in the presence of unsampled populations, it gives good estimates of the rate of migration among sampled populations as well as of the rate of migration into each sampled population from the pooled unsampled populations.


Presence of gamma-aminobutyric acid in embryonic palates of AJ and SWV mouse strains.
The presence of gamma-aminobutyric acid (GABA) in the embryonic palate was sought as a criterion for its role in regulating palate development. GABA was measured by a gas chromatographic-mass spectrometric (GC-MS) method using the heptafluorobutyryl (HFB)-cyclohexyl-GABA derivative, which gave the necessary sensitivity and specificity to measure low levels of GABA in the presence of contaminating substances. GABA was measured in dissected embryonic palates at various times of development in the AJ mouse strain. GABA levels were lower in day 14 AJ palates (0.19 +/- 0.01 nmol/mg protein) than at days 13 (0.28 +/- 0.03) and 15 (0.30 +/- 0.04). Comparable levels were observed in fore- and hindlimbs at day 14, whereas levels were lower in embryonic tongue and higher, as was expected, in embryonic brain. To confirm the presence of GABA in the palate, it was analyzed in growing palate mesenchymal cells in primary and secondary cultures as well as in serum-free medium. In addition, GABA levels were compared in the SWV mouse strain; this strain exhibits a more efficient active uptake mechanism and diazepam produces a higher frequency of cleft palate in this strain than in AJ. SWV contained one and one-half to three times higher concentrations of GABA in excised palates and cultured palate cells than the AJ strain. Furthermore, when GABA levels in skin fibroblasts of the two strains were measured, SWV cells contained 2.7-fold greater GABA than AJ cells. The present results provide additional evidence for the role of GABA in palate development.


Minimally invasive surgical treatment of Bertolotti's Syndrome: case report.
This article aims to provide more insight into the presentation, diagnosis, and treatment of Bertolotti's syndrome, which is a rare spinal disorder that is very difficult to recognize and diagnose correctly. The syndrome was first described by Bertolotti in 1917 and affects approximately 4 to 8% of the population. It is characterized by an enlarged transverse process at the most caudal lumbar vertebra with a pseudoarticulation of the transverse process and the sacral ala. It tends to present with low back pain and may be confused with facet and sacroiliac joint disease. In this case report, we describe a 40-year-old man who presented with low back pain and was eventually diagnosed with Bertolotti's syndrome. The correct diagnosis was made based on imaging studies which included computed tomographic scans, plain x-rays, and magnetic resonance imaging scans. The patient experienced temporary relief when the abnormal pseudoarticulation was injected with a cocktail consisting of lidocaine and steroids. In order to minimize the trauma associated with surgical treatment, a minimally invasive approach was chosen to resect the anomalous transverse process with the accompanying pseudoarticulation. The patient did well postoperatively and had 97% resolution of his pain at 6 months after surgery. As with conventional surgical approaches, a complete knowledge of anatomy is required for minimally invasive spine surgery. This case is an example of the expanding utility of minimally invasive approaches in treating spinal disorders.


HIV-1 reverse transcriptase inhibitors: current issues and future perspectives.
One of the major advances in the recent history of the treatment of HIV infections has been the development of different classes of effective antiretroviral drugs. In particular, the reverse transcriptase (RT) inhibitors still represent the majority of the clinically used anti-HIV drugs and constitute the main backbone of currently employed combinatorial regimens. Highly active antiretroviral combination chemotherapy (HAART), combining RT and protease inhibitors, has proven the most effective approach to treat HIV disease, since it has been shown to markedly suppress viral replication and appearance of drug resistance for a relatively long period. These therapies, however, do not constitute a definitive cure, since they are not able to completely eradicate the virus from the infected individual. Beside drug toxicity problems, the emergence of drug resistance associated with the particular regimen employed further complicates the situation. This review will summarise the most recent achievements, as well as the future directions in the development of novel anti-RT compounds.


Stress analysis of carotid atheroma in transient ischemic attack patients: evidence for extreme stress-induced plaque rupture.
Plaque rupture has been considered to be the result of its structural failure. The aim of this study is to suggest a possible link between higher stresses and rupture sites observed from in vivo magnetic resonance imaging (MRI) of transient ischemic attack (TIA) patients, by using stress analysis methods. Three patients, who had recently suffered a TIA, underwent in vivo multi-spectral MR imaging. Based on plaque geometries reconstructed from the post-rupture status, six pre-rupture plaque models were generated for each patient dataset with different reconstructions of rupture sites to bridge the gap of fibrous cap from original MRI images. Stress analysis by fluid structure interaction simulation was performed on the models, followed by analysis of local stress concentration distribution and plaque rupture sites. Furthermore, the sensitivity of stress analysis to the pre-rupture plaque geometry reconstruction was examined. Local stress concentrations were found to be located at the plaque rupture sites for the three subjects studied. In the total of 18 models created, the locations of the stress concentration regions were similar in 17 models in which rupture sites were always associated with high stresses. The local stress concentration region moved from circumferential center to the shoulder region (slightly away from the rupture site) for a case with a thick fibrous cap. Plaque wall stress level in the rupture locations was found to be much higher than the value in non-rupture locations. The good correlation between local stress concentrations and plaque rupture sites, and generally higher plaque wall stress level in rupture locations in the subjects studied could provide indirect evidence for the extreme stress-induced plaque rupture hypothesis. Local stress concentration in the plaque region could be one of the factors contributing to plaque rupture.


Regulatory processes of hunger motivated behavior.
While food intake and body weight are under homeostatic regulation, eating is a highly motivated and reinforced behavior that induces feelings of gratification and pleasure. The chemical senses (taste and odor) and their evaluation are essential to these functions. Brainstem and limbic glucose-monitoring (GM) neurons receiving neurochemical information from the periphery and from the local brain milieu are important controlling hunger motivation, and brain gut peptides have a modulatory role on this function. The hypothalamic and limbic forebrain areas are responsible for evaluation of reward quality and related emotions. They are innervated by the mesolimbic dopaminergic system (MLDS) and majority of GM neurons are also influenced by dopamine. Via dopamine release, the MLDS plays an essential role in rewarding-reinforcing processes of feeding and addiction. The GM network and the MLDS in the limbic system represent essential elements in the neural substrate of motivation.


[Pathomorphologic changes in the testes of rats fed gamma ray irradiated products].
A study was made of morphological changes in the testes of 120 rats fed for 20 months only food exposed to gamma-rays. Pronounced alterative-dystrophic disorders up to foci of coagulation necrosis and aspermatogenesis as well as compensatory-adaptation processes such as proliferation of Leydig cells were found. A direct correlation was disclosed between the severity of structural alterations in the testes and the dose of gamma-rays to which the products used for animals' feeding had been exposed. A suggestion is made about the autoimmune mechanism of structural alterations discovered.


Kindler syndrome. Clinical and ultrastructural findings.
Kindler syndrome is a genodermatosis that combines clinical features of hereditary epidermolysis bullosa and poikiloderma congenitale. The ultrastructural level of blister formation has not been well characterized. Two brothers with Kindler syndrome had a history of primarily acral blistering since infancy as well as photosensitivity. Blister formation was found through the basal layer. Marked tonofilament clumping was found in intact keratinocytes adjacent to the blisters. The younger brother (aged 21 years) had actinic keratoses, which have not been previously described in Kindler syndrome. The findings of basal layer separation in both spontaneous and induced blisters in Kindler syndrome suggest this is the true level of blister formation. The finding of actinic keratoses in a young patient with Kindler syndrome suggests that some patients may be at increased risk for early solar-induced skin disease. The presence of clumped tonofilaments in keratinocytes adjacent to blistered areas suggests an abnormality of keratin 5 or 14 could be present and may play a role in blister formation in patients with Kindler syndrome.


Management of Renin-Angiotensin-Aldosterone System blockade in patients admitted to hospital with confirmed coronavirus disease (COVID-19) infection (The McGill RAAS-COVID- 19): A structured summary of a study protocol for a randomized controlled trial.
The aim of the RAAS-COVID-19 randomized control trial is to evaluate whether an upfront strategy of temporary discontinuation of renin angiotensin aldosterone system (RAAS) inhibition versus continuation of RAAS inhibition among patients admitted with established COVID-19 infection has an impact on short term clinical and biomarker outcomes. We hypothesize that continuation of RAAS inhibition will be superior to temporary discontinuation with regards to the primary endpoint of a global rank sum score. The global rank sum score has been successfully used in previous cardiovascular clinical trials. This is an open label parallel two arm (1,1 ratio) randomized control superiority trial of approximately 40 COVID-19 patients who are on chronic RAAS inhibitor therapy. Adults who are admitted to hospital within the McGill University Health Centre systems (MUHC) including Royal Victoria Hospital (RVH), Montreal General Hospital (MGH) and Jewish General Hospital (JGH) and who are within 96 hours of COVID-19 diagnosis (confirmed via PCR on any biological sample) will be considered for the trial. Of note, the initial protocol to screen and enrol within 48 hours of COVID-19 diagnosis was extended through an amendment, to 96 hours to increase feasibility. Participants have to be 18 years or older and would have to be on RAAS inhibitors for at least a month to be considered eligible for the study. Additionally, RAAS inhibitors should not have been held for more than 48 hours before randomization. A list of inclusion and exclusion criteria can be found in the full protocol document. In order to prevent heart failure exacerbation, patients with reduced ejection fraction were excluded from the trial. Once a patient is admitted on the ward with a diagnosis of COVID-19, we will confirm with the treating physician if the participant is suitable for the RAAS-COVID trial and meets all the inclusion and exclusion criteria. If the patient is eligible and informed consent has been obtained we will collect data on sex, age, ethnicity, past medical history and list of medications (e.g. other anti-hypertensives or anticoagulants), for further analysis. All the study participants will be randomized to a strategy of temporarily holding the RAAS inhibitor [intervention] versus continuing the RAAS inhibitor [continued standard of care]. Among participants who are randomized to the intervention arm, alternative guide-line directed anti-hypertensive medication will be provided to the treating physician team (detail in study protocol). In the intervention arm RAAS inhibitor will be withheld for a total of 7 days with the possibility of the withdrawn medication being initiated at any point after day 7 or on the day of discharge. The recommendation for re-initiating the withdrawn medication will be made to the treating physician. The re-initiation of these therapies are according to standard convention and follow-up as per Canadian guidelines. Additionally, the date of restarting the withdrawn medication or whether the medication was re-prescribed on discharge or not, will be collected. This will be used to conduct a sensitivity analysis. Furthermore, biomarkers such as troponin, c-reactive protein (CRP) and lymphocyte count will be assessed during the same time period. Samples will be collected on randomization, day 4 and day 7. PRIMARY ENDPOINT: In this study the primary end point is a global rank score calculated for all participants, regardless of treatment assignment ( score from 0 to 7). Please refer to table 4 in the full protocol. In the context of the current trial, it is estimated that death is the most meaningful endpoint, and therefore has the highest score ( score of 7). This is followed by admission to ICU, the need for mechanical ventilation etc. The lowest scores ( score of 1) are assigned to biomarker changes (e.g. change in troponin, change in CRP). This strategy has been used successfully in cardiovascular disease trials and therefore is applicable to the current trial. The primary endpoint for the present trial is assessed from baseline to day 7 (or discharge). Participants are ranked across the clinical and biomarker domains. Lower values indicate better health (or stability). Participants who died during the 7th day of the study will be ranked based on all events occurring before their death and also including the fatal event in the score. Next, participants who did not die but were transferred to ICU for invasive ventilation will be ranked based on all the events occurring before the ICU entry and also including the ICU admission in the score. Those participants who did not die were not transferred to ICU for invasive ventilation, will be ranked based on the subsequent outcomes. The mean rank score will then be compared between groups. In this scheme, a lower mean rank score indicates greater overall stability for participants. Secondary endpoints : The key secondary endpoints are the individual components of the primary components and include the following: death, transfer to ICU primarily for invasive ventilation, transfer to ICU for other indication, non-fatal MACE ( any of following, MI, stroke, acute HF, new onset Afib), length of stay > 4 days, development of acute kidney injury ( > 40% decline in eGFR or doubling of serum creatinine), urgent intravenous treatment for high blood pressure, 30% increase in baseline high sensitivity troponin, 30% increase in baseline BNP, increase in CRP to > 30% in 48 hours and lymphocyte count drop> 30%. We will also look at the World Health Organization (WHO) ordinal scale for clinical improvement (in COVID-19) in our data. In this scale death will be assigned the highest score of 8. Patients with no limitation of activity will be assigned a score of 1 which indicates overall more stability (3). Additionally, we will evaluate the potential effects of discontinuing RAAS inhibition on alternative schedules (longer/shorter than 7 days, intermittent discontinuation) using a mechanistic mathematical model of COVID-19 immunopathology calibrated to data collected from our patient cohort. In particular, we will assess the impact of alternative schedules on primary and secondary endpoints including increases to baseline CRP and lymphocyte counts. Participants will be randomized in a 1:1 ratio. Randomization will be performed within an electronic database system at the time of enrolment using a random number generator, an approach that has been successfully used in other clinical trials. Neither participant, study team, or treating team will be blinded to the intervention arm. This is an open label study with no blinding. The approximate number of participants required for this trial is 40 patients (randomized 1:1 to continuation versus discontinuation of RAAS inhibitors). This number was calculated based on previous rates of outcomes for COVID-19 in the literature (e.g. death, ICU transfer) and statistical power calculations. Protocol number: MP-37-2021-6641, Version 4: 01-10-2020. Trial start date September 1st 2020 and currently enrolling participants. Estimated end date for recruitment of participants : July 2021. Estimated end date for study completion: September 1st 2021. Trial registration: ClincalTrials.gov : NCT04508985 , date of registration: August 11th , 2020 FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


[Indication procedure: how difficult we make things!].
The Interim Advice of the Centrale Raad voor de Volksgezondheid, Concerning the procedure of indication in nursing homes, is directed at coordination with existing admitting procedures in homes for the elderly, but it does not take into consideration the differences between both provisions. The advice disregards the, in the meantime growing, practice of a geriatric examination which is closely connected to admission in nursing homes.


Precise mapping of increased sialylation pattern and the expression of acute phase proteins accompanying murine tumor progression in BALB/c mouse by integrated sera proteomics and glycomics.
Inbred BALB/c mouse implanted with murine tumors serves as an attractive model system for the studies of cancer biology in immuno-competent individuals. It is anticipated that tumor progression would induce notable pathophysiological consequences, some of which manifested as alteration in serum proteomic and glycomic profiles. Similar to sera derived from human cancer patients and immuno-compromised mice bearing human tumors, we show in this work that BALB/c mice of the same genetic background but bearing two distinct tumor origins both exhibited elevated expression levels of acute phase proteins including haptoglobin and serum amyloid P protein, in response to tumor progression. Such common traits are generally not informative nor qualifying as biomarkers. Additional mass spectrometry (MS)-based glycomic mapping nevertheless detected distinctive changes of sialylation pattern on the complex type N-glycans. MALDI MS/MS sequencing afforded a facile but definitive identification of an increase in internal Neu5Gcalpha2-6 sialylation on the GlcNAc of the Neu5Gc2-3Gal1-3GlcNAc terminal sequence as a common feature whereas a substitution of Neu5Gc by Neu5Ac was found to be induced by colonic but not breast tumor. A more pronounced change was similarly detected on N-glycans derived from ascitic fluids representing late tumor progression stages. We next demonstrated that such distinct change in glycotope expression can be localized to a particular protein carrier by LC-MS/MS analysis of glycopeptides. Serotransferrin was identified as one such abundant serum glycoprotein, which changed significantly not in protein expression level but in terminal glycosylation pattern.


[Multiple traumatic intracranial hematomas in children].
Of 32 children with traumatic intracranial hematomas 11 had multiple hematomas. The latter were characterized by a faster development of the compression syndrome with signs of intracranial hypertension and secondary stem symptoms. "Clear intervals" were more frequent in children than adults. Computer-aided tomographic examination is a valuable method in the diagnosis of multiple hematomas.


Behavioral considerations for engaging youth in HIV clinical research.
From both scientific and ethical perspectives, it is important that youth be enrolled in biomedical HIV prevention clinical trials. At the same time, adolescents, as minors, are considered a vulnerable population requiring particular attention to the reduction of potential harm associated with participation in such trials. In this article, we review the evidence supporting enrollment of youth in HIV clinical trials, including data on HIV infection rates, sexual behavior, and cognitive, psychosocial, and neurophysiological development. Next, we address the potential risks associated with clinical trial participation, with a focus on the concept of preventive misconception, the tendencies to (1) overestimate the probability of assignment to the experimental condition, as opposed to the placebo, and (2) assume that the experimental intervention is efficacious. Finally, we discuss targeted interventions to reduce preventive misconception and the importance of developing and testing adolescent-friendly risk-reduction interventions that are tailored to the structure and time frame of a biomedical HIV prevention clinical trial. The very issues that make inclusion of youth in HIV prevention clinical trials necessary also demand that particularly intensive efforts be made to protect participating minors from the harm that could accrue from a clinical trial.


A Biological-Realtime Neuromorphic System in 28 nm CMOS Using Low-Leakage Switched Capacitor Circuits.
A switched-capacitor (SC) neuromorphic system for closed-loop neural coupling in 28 nm CMOS is presented, occupying 600 um by 600 um. It offers 128 input channels (i.e., presynaptic terminals), 8192 synapses and 64 output channels (i.e., neurons). Biologically realistic neuron and synapse dynamics are achieved via a faithful translation of the behavioural equations to SC circuits. As leakage currents significantly affect circuit behaviour at this technology node, dedicated compensation techniques are employed to achieve biological-realtime operation, with faithful reproduction of time constants of several 100 ms at room temperature. Power draw of the overall system is 1.9 mW.


Quantitative metabolomics of a xylose-utilizing Saccharomyces cerevisiae strain expressing the Bacteroides thetaiotaomicron xylose isomerase on glucose and xylose.
The yeast Saccharomyces cerevisiae cannot utilize xylose, but the introduction of a xylose isomerase that functions well in yeast will help overcome the limitations of the fungal oxido-reductive pathway. In this study, a diploid S. cerevisiae S288c[2n YMX12] strain was constructed expressing the Bacteroides thetaiotaomicron xylA (XI) and the Scheffersomyces stipitis xyl3 (XK) and the changes in the metabolite pools monitored over time. Cultivation on xylose generally resulted in gradual changes in metabolite pool size over time, whereas more dramatic fluctuations were observed with cultivation on glucose due to the diauxic growth pattern. The low G6P and F1,6P levels observed with cultivation on xylose resulted in the incomplete activation of the Crabtree effect, whereas the high PEP levels is indicative of carbon starvation. The high UDP-D-glucose levels with cultivation on xylose indicated that the carbon was channeled toward biomass production. The adenylate and guanylate energy charges were tightly regulated by the cultures, while the catabolic and anabolic reduction charges fluctuated between metabolic states. This study helped elucidate the metabolite distribution that takes place under Crabtree-positive and Crabtree-negative conditions when cultivating S. cerevisiae on glucose and xylose, respectively.


Some pharmacological aspects of xanthines in asthma.
Xanthines, best represented by theophylline, constitute a pharmacological principle of considerable age and importance in the treatment of asthma. Historical aspects of xanthines, effects of these compounds in the lung, and xanthine drug research are briefly reviewed. Data are presented on the active metabolite of theophylline, on relaxant effects of theophylline in large and small human respiratory airways, and on anti-edema effects of theophylline.


Access to Continuing Education for Critical Care Nurses in Rural or Remote Settings.
Caring for the critically ill does not occur solely in large medical centers or urban areas. Nurses practicing in rural or remote settings practice as nurse generalists, caring for a wide range of patients, including those needing critical care. As a nurse generalist, the need for a wide variety of skill sets challenges nurses in rural areas to maintain current practice through continuing education across many content areas. They also may not come in contact with critically ill patients or emergent situations as often as their urban counterparts, which can make remaining comfortable with more advanced skills difficult. Because nurses working in rural or remote areas may care for critically ill patients less often, the need to remain vigilant in pursuit of continuing education is increased; however, access to continuing education can be problematic because of geographic isolation, difficulty getting time off from work, limited financial resources, and perceived lack of applicable topics. With advances in technology, critical care nurses working in rural areas have more options for continuing education, which is crucial for maintaining a skilled nursing workforce in rural health care facilities. This article addresses challenges critical care nurses working in remote or rural areas face in pursuing continuing education and provides information about available educational opportunities.


The mating type locus (MAT) and sexual reproduction of Cryptococcus heveanensis: insights into the evolution of sex and sex-determining chromosomal regions in fungi.
Mating in basidiomycetous fungi is often controlled by two unlinked, multiallelic loci encoding homeodomain transcription factors or pheromones/pheromone receptors. In contrast to this tetrapolar organization, Cryptococcus neoformans/Cryptococcus gattii have a bipolar mating system, and a single biallelic locus governs sexual reproduction. The C. neoformans MAT locus is unusually large (>100 kb), contains >20 genes, and enhances virulence. Previous comparative genomic studies provided insights into how this unusual MAT locus might have evolved involving gene acquisitions into two unlinked loci and fusion into one contiguous locus, converting an ancestral tetrapolar system to a bipolar one. Here we tested this model by studying Cryptococcus heveanensis, a sister species to the pathogenic Cryptococcus species complex. An extant sexual cycle was discovered; co-incubating fertile isolates results in the teleomorph (Kwoniella heveanensis) with dikaryotic hyphae, clamp connections, septate basidia, and basidiospores. To characterize the C. heveanensis MAT locus, a fosmid library was screened with C. neoformans/C. gattii MAT genes. Positive fosmids were sequenced and assembled to generate two large probably unlinked MAT gene clusters: one corresponding to the homeodomain locus and the other to the pheromone/receptor locus. Strikingly, two divergent homeodomain genes (SXI1, SXI2) are present, similar to the bE/bW Ustilago maydis paradigm, suggesting one or the other homeodomain gene was recently lost in C. neoformans/C. gattii. Sequencing MAT genes from other C. heveanensis isolates revealed a multiallelic homeodomain locus and at least a biallelic pheromone/receptor locus, similar to known tetrapolar species. Taken together, these studies reveal an extant C. heveanensis sexual cycle, define the structure of its MAT locus consistent with tetrapolar mating, and support the proposed evolutionary model for the bipolar Cryptococcus MAT locus revealing transitions in sexuality concomitant with emergence of a pathogenic clade. These studies provide insight into convergent processes that independently punctuated evolution of sex-determining loci and sex chromosomes in fungi, plants, and animals.


Patient characteristics and outcomes of a home mechanical ventilation program in a developing country.
There are limited data on home mechanical ventilation (HMV) in developing countries. This study aimed to describe the patient characteristics, feasibility, and outcomes of an HMV program at a university hospital in Thailand. Data were collected on all patients who were discharged with HMV between October 2014 and August 2015 at Thammasat University Hospital. Twelve patients (eight men and four women) underwent HMV. They were aged 71.5 ± 17.6 years; mean ± standard deviation. Indications for HMV were 6 neurologic diseases (4 amyotrophic lateral sclerosis, 1 multiple system atrophy, and 1 stroke), 2 chronic obstructive pulmonary disease (COPD), 1 tracheomalacia, and 3 combined neurologic diseases and respiratory diseases (2 stroke and COPD, 1 stroke and tracheomalacia). The duration of follow-up was 799.5 ± 780.5 days. The ratio of family income to cost of HMV usage was 77.2:1 ± 5.5:1. All patients had tracheostomies. Modes of HMV were biphasic positive airway pressure (66.7%), pressure-controlled ventilation (16.7%), pressure-support ventilation (8.3%), and volume-controlled ventilation (8.3%). Complications occurred in ten patients (83.3%), including tracheobronchitis (20 events) and ventilator-associated pneumonia (12 events). Overall mortality was 41.7% (5/12 patients), including two patients who died due to ventilator-associated pneumonia. There were no instances of ventilator malfunction. HMV is feasible for patients with neurological diseases and COPD in a developing country. The relatively high rate of complications indicates the need for more comprehensive clinical services for chronic ventilator-dependent patients in this setting.