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Abstract Background Linezolid is an effective, but toxic anti-tuberculosis drug that is currently recommended for the treatment of drug-resistant tuberculosis. Improved oxazolidinones should have a better safety profile, while preserving efficacy. Delpazolid is a novel oxazolidinone developed by LegoChem Biosciences Inc. that has been evaluated up to phase 2a clinical trials. Since oxazolidinone toxicity can occur late in treatment, LegoChem Biosciences Inc. and the PanACEA Consortium designed DECODE to be an innovative dose-ranging study with long-term follow-up for determining the exposure–response and exposure–toxicity relationship of delpazolid to support dose selection for later studies. Delpazolid is administered in combination with bedaquiline, delamanid and moxifloxacin. Methods Seventy-five participants with drug-sensitive, pulmonary tuberculosis will receive bedaquiline, delamanid and moxifloxacin, and will be randomized to delpazolid dosages of 0 mg, 400 mg, 800 mg, 1200 mg once daily, or 800 mg twice daily, for 16 weeks. The primary efficacy endpoint will be the rate of decline of bacterial load on treatment, measured by MGIT liquid culture time to detection from weekly sputum cultures. The primary safety endpoint will be the proportion of oxazolidinone class toxicities; neuropathy, myelosuppression, or tyramine pressor response. Participants who convert to negative liquid media culture by week 8 will stop treatment after the end of their 16-week course and will be observed for relapse until week 52. Participants who do not convert to negative culture will receive continuation phase treatment with rifampicin and isoniazid to complete a six-month treatment course. Discussion DECODE is an innovative dose-finding trial, designed to support exposure-response modelling for safe and effective dose selection. The trial design allows assessment of occurrence of late toxicities as observed with linezolid, which is necessary in clinical evaluation of novel oxazolidinones. The primary efficacy endpoint is the change in bacterial load, an endpoint conventionally used in shorter dose-finding trials. Long-term follow-up after shortened treatment is possible through a safety rule excluding slow-and non-responders from potentially poorly performing dosages. Trial registration DECODE was registered in ClinicalTrials.gov before recruitment start on 22 October 2021 (NCT04550832).

Background. Burnout during registrar training is high, especially in resource-limited settings where stressors are intensified. Burnout leads to decreased quality of life for doctors, poor job and patient satisfaction, and difficulty retaining doctors.Objectives. Primary: to measure burnout among registrars working at Princess Marina Hospital in Gaborone, Botswana. Secondary: to determine factors contributing to burnout and identify potential wellness interventions.Methods. The validated Maslach Burnout Inventory was used to measure the degree of emotional exhaustion, depersonalisation and personal accomplishment. Work-related difficulties and potential wellness interventions were explored through multiple-choice and open-ended questions.Results. Of 40 eligible registrars, 20 (50%) completed the survey. High levels of burnout were reported for emotional exhaustion in 65% (13/20), depersonalisation in 45% (9/20), and personal accomplishment in 35% (7/20) of registrars. A high degree of burnout was reported by 75% (15/20) of registrars in one or more domains. In the previous 7 days, registrars worked an average of 77 hours, took 1.5 overnight calls, slept 5.7 hours per night, and 53% (10/19) had ≥1 of their patients die. Five (25%) registrars considered leaving Botswana to work in another country, which correlated with those with the highest degree of burnout. The most common frustrations included insufficient salary and limited medical resources. Suggested interventions included improved mentorship and wellness lectures.Conclusions. There is a high degree of burnout, especially emotional exhaustion, among registrars. Encouragingly, most registrars have a desire to work in Botswana after training. Future research on improving registrar wellness in low-resource settings is urgently needed.

This study describes the predominantly tropical, subtidal seaweed populations growing on rhodoliths between 4 and 18 m depth in the southern part of Espírito Santo State (Brazil). Qualitative and quantitative sampling revealed species-rich algal communities, comprising 167 species. Three species of rhodophytes represent new records for the Brazilian marine flora (Lithothamnion muelleri, Scinaia aborealis, and Mesophyllum engelhartii). Marked seasonal differences in fleshy algal species composition and abundance were related to seasonal instabilities caused by winter-storm disturbance over the rhodolith beds. In relation to depth, rhodolith density appears to be an important factor for the variation in the abundance of fleshy algae. The rhodolith community is composed of at least seven nongeniculate crustose coralline algal species. Rhodolith beds in southern Espírito Santo State, in an area of 150 km2, provide an important habitat for epibenthic communities, supporting 25% of the known macroalgal species richness along the Brazilian coast.

One kind of autonomous vehicle that can take instructions from the driver by reading their electroencephalogram (EEG) signals using a Brain-Computer Interface (BCI) is called a Brain-Controlled Vehicle (BCV). The operation of such a vehicle is greatly affected by how well the BCI works. At present, there are limitations on the accuracy of BCI recognition, the number of distinguishable command categories, and the execution duration of command recognition. Consequently, vehicles that are exclusively controlled by EEG signals demonstrate suboptimal control performance. To address the difficulty of improving the control capabilities of brain-controlled cars while maintaining BCI performance, a fuzzy logic-based technique called as Fuzzy Brain-Control Fusion Control is introduced. This approach uses Fuzzy Discrete Event System (FDES) supervisory theory to verify the accuracy of the driver's brain-controlled directives. Concurrently, a fuzzy logic-based automatic controller is developed to generate decisions automatically in accordance with the present state of the vehicle via fuzzy reasoning. The final decision is then reached through the application of secondary fuzzy reasoning to the accuracy of the driver's instructions and the automated decisions to make adjustments that are more consistent with human intent. A clever BCI gadget known as the Consistent State Visual Evoked Potential (SSVEP) is utilized to show the viability of the proposed technique. We recommend that additional research should be conducted at this time to confirm that our recommended system may further improve the control execution of BCI-fueled cars, regardless of whether BCIs have special limitations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Introduction Post-pandemic alcohol consumption is on the rise due to people starting to adapt themselves to the practice of consuming alcoholic beverages at home. In addition to the direct effects of intoxication and addiction, estimates suggest that alcohol contributes to approximately 20-30% of global cases of oesophagal cancer, liver cancer, cirrhosis of the liver, homicide, epilepsy, and motor vehicle accidents. In India, one-fifth of alcohol consumers were found to be alcohol dependent. The study was done with the primary objective of finding out the prevalence of alcohol dependence among alcohol users and exploring the reasons for alcohol dependence among alcohol users in an urban area of Chengalpattu District, Tamil Nadu. Methodology The study design is an explanatory sequential mixed-methods study. It was done among 624 adult male alcohol consumers in the Chennai district, selected by the cluster sampling method in a community setting. The Alcohol Use Disorders Identification Test (AUDIT) was used to diagnose alcohol dependence. Using the purposive sampling method, in-depth interviews were conducted among 24 alcohol-dependent people to explore and understand their experiences, identify common themes, and provide insights into the problem. Quantitative data were analysed using Statistical Package for Social Sciences (SPSS) version 26 (IBM Corp., Armonk, NY), and qualitative data were analysed using deductive content analysis using Qualcoder software. Results The mean age of the study participants was 38±7 years. Among current alcohol consumers, 16.9% (106/624) were found to be suffering from alcohol dependence. The significant predictors of alcohol dependence were found to be unskilled occupation (adjusted odds ratio [AOR] = 2.09), having suicidal ideation (AOR = 2.4), alcohol consumption by family members (AOR = 1.90), depression (AOR = 3.98), drinking pattern-affected interpersonal relationships (AOR = 2.29), and not receiving health education about alcohol use in school/college (AOR = 1.74). The major themes and codes identified among alcohol dependents were factors related to mental health, physical health, and social factors. Conclusion This study provides essential points of reference for policymakers and primary care physicians to develop prevention strategies for people to understand and overcome the problem of alcohol addiction, and it also sheds light on the burden of alcohol dependence and their lived experiences., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Institutional Human Ethical Committee of Sree Balaji Medical College issued approval SBMCH/IHEC/2023/96. Ethical approval was obtained from the Institutional Human Ethical Committee of a tertiary medical college in Chengalpattu district (Approval number: SBMCH/IHEC/2023/96, Approval Date: 11.06.23). Informed consent was obtained from each study participant before enrolment. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Baskaran et al.)

In the Chilean pediatric population, to date, there is no validated screening instrument for sleep-related breathing disorders (SRBD)., Objective: to develop and validate a cross-cultural adaptation of the Pediatric Sleep Questionnaire - sleep-related breathing disorder scale (PSQ-SRBD), by creating the Chilean Spanish version (PSQ-CL)., Patients and Method: The PSQ-SRBD was translated from English into Chilean Spanish, obtaining the PSQ-CL, which was subsequently validated. Internal consistency was determined through Cronbach's alpha coefficient in 26 children with obstructive sleep apnea confirmed by polysomnography and in 112 controls. Reliability was obtained through the test-retest method., Results: In the pilot group, the overall internal consistency of the PSQ- CL through Cronbach's alpha was 0.71 and the internal consistency was 0.653, 0.566, and 0.808 in subscales A, B, and C, respectively. The discrimination capacity of the PSQ-CL questionnaire established through ROC analysis was 81%, determining a cut-off score of 0.227 with a sensitivity of 73.08% and a specificity of 77.68%., Conclusions: The PSQ-CL is a suitable instrument for screening sleep-disordered breathing in Chilean children. This instrument may be useful in clinical practice and epidemiological research in the Chilean pediatric population and could be used for multicenter studies.

Background: Microcrystalline cellulose (MCC) is a novel organic material developed by one of the authors in this study. When MCC was incorporated with conventionally available denture base resin, it demonstrated increased flexural strength and flexural modulus. However, it was speculated that because the material is organic, it can promote the growth of Candida . The purpose of this study is to evaluate the Candida albicans biofilm formation on polymethyl methacrylate (PMMA) denture base resin incorporated with MCC., Materials and Methods: MCC is an organic material extracted from the oil palm empty fruit bunch (OPEFB). The growth of C. albicans and biofilm formation in three test groups were compared by biofilm assay and imaging techniques like microscopy (by safranin staining) and scanning electron microscopy. The three test groups were comprised of MCC-reinforced PMMA containing OPEFB fibers of 50-micrometer thickness at 5% weight reinforcement, conventionally and commercially available heat cure PMMA, and an empty well to assess any discrepancies from the environment., Results: The test groups showed increased biofilm formation by C. albicans compared to commercially and conventionally available heat cure PMMA. Reinforcement with MCC showed higher biofilm formation of 1.43 times higher compared to conventional PMMA. Biofilms formed by Candida albicans on MCC-reinforced PMMA appeared heterogeneous in structure, comprised of yeast cells and hyphae, surrounded by a higher density of polysaccharide extracellular matrix material compared to that of conventionally available heat cure PMMA., Conclusion: Biofilm formation is increased in denture base resin incorporated with MCC. More investigation is warranted to study the antifungal efficacy of the addition of antifungal agents to the reinforced denture base resin., Competing Interests: Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, S et al.)

Competing Interests: The authors report no conflicts of interest.

Background: Pathology, a foundational yet challenging subject in medical education, is marked by its extensive content and intricate concepts. These complexities often pose a significant learning barrier for students, who must not only comprehend but also effectively apply this knowledge in their clinical practice., Objective: This study aimed to investigate the impact of utilizing cartoons as a supplementary educational tool in pathology. Specifically, it focused on assessing whether incorporating cartoons into the learning process would enhance students' understanding, memory retention, and ability to recall complex topics, thereby augmenting the effectiveness of traditional teaching methodologies., Materials and Methods: Conducted from June to September 2022, this experimental study involved 106 second-year MBBS (Bachelor of Medicine and Bachelor of Surgery) students. Participants were split into two groups: the "traditional group," which received standard interactive large-group teaching, and the "combination group," which benefited from both the standard teaching and additional cartoon-based instruction. The study focused on two selected chapters of the pathology curriculum. After completing the first chapter, the groups were crossed over for the second chapter. Evaluation of the students' learning was conducted through post-learning assessments using multiple-choice questions (MCQs)., Results: The combination group, which received both traditional and cartoon-based teaching, showed a significant improvement in their assessment scores compared to the traditional group. This improvement was consistent in both assessments conducted (t(102) = 8.41, p < .001 and t(99) = 6.85, p < .001). Additionally, feedback from the students through a post-learning survey indicated a strong preference for the use of cartoons. The majority of students agreed that cartoons facilitated a better understanding and retention of complex pathology topics (χ² = 130.9, p < 0.001)., Conclusion: The incorporation of cartoons as a supplementary learning tool in pathology teaching shows promising results. This innovative approach not only complements but also enhances the traditional teaching methods, leading to improved comprehension, retention, and recollection of complex subjects among medical students. The study highlights the potential of cartoons in revolutionizing medical education, especially in teaching challenging subjects like pathology., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Kathiah et al.)

Patients placed in a prone position due to COVID-19 present a more severe disease, longer stay in ICU and have more significant complications associated with positioning than other patients., Aim: To identify social, health, and hospital factors associated with mortality and functional recovery in activities of daily living (ADL) in patients admitted to the ICU due to COVID-19 who were in the prone position., Methods: We conducted a prospective observational study between June 2021 and March 2022 at the San José Clinical Hospital. The study included people 18 years of age or older admitted to the ICU due to COVID-19 who were in the prone position. Using logistic regression, we searched for factors associated with the risk of in-hospital mortality. Using linear regression, we also analyzed factors associated with functional recovery in ADL at discharge and post-discharge., Results: In 85 patients included, being men, older age, history of mental health, cardiorespiratory disease, and living in a commune of high social priority were associated with a higher risk of in-hospital mortality. A greater number of days of hospitalization was associated with less functional recovery at discharge and post-discharge. In contrast, more cycles in prone position during hospitalization and higher education level were associated with greater post-discharge functional recovery., Conclusions: The survival and functional prognosis of patients with severe COVID-19 can be predicted early through a biopsychosocial evaluation of their characteristics. Intrahospital actions could improve functional recovery in ADL in the short term in critically ill patients surviving COVID-19.

Objectives: The objective is to assess and compare whether AdvanSync gives better skeletal, dental, and soft tissue outcomes than Forsus fixed functional appliance in post-pubertal skeletal Class II malocclusion patients., Methodology: A prospective study was conducted using 3D-CBCT of patients taken before and after fixed functional appliance therapy. The sample consisted of 16 patients divided into two groups: Group 1 was treated with Forsus and group 2 with AdvanSync appliance. All subjects were in their post-pubertal growth phase. Treatment changes were evaluated between the study groups using 12 angular and 14 linear parameters. The data were subjected to statistical analysis., Results: Statistically significant changes in SNB (P:0.04) and ANB (P:0.01) in Forsus appliance and AdvanSync (SNB, P:008), (ANB, P: <0.001) were found between the pre and post-fixed functional appliance protocol. The effective mandibular length increased in both groups Forsus(P-value: 0.01) and AdvanSync (P-value: 0.01). Group 1 resulted in lower incisor proclination and intrusion, a reduction in an overbite, whereas group 2 resulted in lower incisor proclination, upper incisor extrusion, and retroclination. Both groups showed significant improvement in the molar relation and overjet. An increase in the total facial convexity was shown in group 1; group 2 showed an increase in the facial convexity and total facial convexity. There was no statistically significant difference between the effects of the Forsus and Advansync appliance groups in the post-fixed functional period. Both groups presented similar results., Conclusion: Forsus and Advansync appliances are effective with similar results in normalizing skeletal Class II malocclusion in post-pubertal patients. There was no statistically significant difference between the groups using Forsus or AdvanSync. The changes were minimal - mandibular skeletal effects, majorly dentoalveolar effects, and mild soft tissue effects., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, M et al.)

Plant-mediated nanoparticle synthesis is a revolutionary technique with numerous applications in fields, such as agriculture, food processing, and medicine. This study reports that Punica granatum leaf extract is capable of the green and eco-friendly synthesis of silica nanoparticles that provides a simple, cost-effective, and efficient methodology. P. granatum leaf extract was employed as a capping and stabilizing agent for the formation of silica nanoparticles, which were synthesized by a biological method using tetra ethyl ortho silicate. Biosynthesized silica nanoparticles are characterized by X-ray diffraction analysis, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and Fourier transform infrared spectroscopy. All the analyses and characterization determined that the particles were spherical in shape and amorphous in nature with an average size of 12 nm. P. granatum-assisted silica nanoparticles were tested for antibacterial activity by a well-diffusion method against two-gram negative bacterial pathogens (E. coli and Salmonella sp.). The antibacterial studies prove that P. granatum-assisted silica nanoparticles have good antibacterial properties. These studies will help us find a new nano-drug or medicine against multidrug-resistant bacteria., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background The aim of this study is to assess the prevalence of metabolic syndrome in tuberculosis-associated obstructive airway disease (TOPD) patients, as well as the association of its components with the severity of airflow obstruction. Methodology In this cross-sectional analytical study, we evaluated the clinical profile, spirometry, waist circumference, blood pressure, lipid profile, fasting plasma glucose, and the association of each component with the severity of airflow obstruction. Results The prevalence of metabolic syndrome in TOPD was found to be was 25.77% (95% confidence interval = 18.11-35.28) among study participants. Reduced high-density lipoprotein was the deranged component and was associated with increased severity in Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage II compared to GOLD stage IV. Conclusions The prevalence of metabolic syndrome in TOPD has a severe impact on patients' treatment, outcomes, and complications. However, in our study, tuberculosis-associated metabolic syndrome was the same as the general population. Low high-density lipoprotein levels were associated with the severity of the airflow obstruction., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, MP et al.)

In this work, a comparative analysis of Fenton, Fe 2+ /NaOCl and Fe 2+ /(NH 4 ) 2 S 2 O 8 assisted advanced oxidation processes was carried out to study its efficacy in enhancing tannery chemical sludge dewaterability. Experimental results showed that, after conditioning of sludge at optimum pH, oxidant and catalyst dosage, capillary suction time (CST) was decreased by 75 ± 5 % and moisture content of sludge cake (M.C cake ) was reduced to 74 ± 2 % for all processes at 10 min reaction time. Among the three processes, Fe 2+ /(NH 4 ) 2 S 2 O 8 process was noted to have larger impact on the breakdown of sludge cells, which is reflected in terms of its highest amount of total organic carbon, protein and total dissolved solids present in the separated sludge fractions. In addition, Fe 2+ /(NH 4 ) 2 S 2 O 8 process was also observed to leach out largest fraction say 73.3 % of chromium present in the raw sludge into the treated sludge filtrate. The investigation carried out through zeta potential and scanning electron microscopy reveals the role of a two-step mechanism, namely oxidation followed by coagulation in enhancing the sludge dewaterability. Further studies on the possibility of utilizing dried treated sludge as biomass fuel could be carried out., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Streptomyces belongs to the Actinomycetes group of bacteria which are gram-positive non acid-fast bacilli, widely recognised for their potential to produce antimicrobials active against bacterial, mycobacterial, parasitic and fungal infections. They commonly cause cutaneous infections following traumatic inoculation. Visceral infections are relatively rare and limited to immunocompro-mised hosts. We describe a case of Streptomyces pneumonia in a healthy immunocompetent female, who when investigated for voluntary kidney donation, resulted in the isolation of Streptomyces species from bronchial wash cultures. Streptomyces, a potential pathogen in immunocompetent hosts is frequently underdiagnosed. Once isolated, both physicians and microbiologists should pay attention to differentiate true infection from contamination.

Netto, M. S. B., Lima, W. G., Correia, K. C., da Silva, C. F. B., Thon, M., Martins, R. B., Miller, R. N. G., Michereff, S. J., and Câmara, M. P. S. 2016. Analysis of phylogeny, distribution, and pathogenicity of Botryosphaeriaceae species associated with gummosis of Anacardium in Brazil, with a new species of Lasiodiplodia. We identified Botryosphaeriaceae species associated with gummosis on Anacardium in Brazil. Isolates were sampled and identified on the basis morphology and phylogeny, through analysis of a partial translation elongation factor 1-α sequence, ribosomal DNA internal transcribed spacers, and β-tubulin gene sequence. Ten taxa were identified, namely, Lasiodiplodia brasiliense, L. euphorbicola, L. gonubiensis, L. iraniensis, L. jatrophicola, L. gravistriata sp. nov., L. pseudotheobromae, L. theobromae, Neofusicoccum batangarum, and Pseudofusicoccum stromaticum. Lasiodiplodia theobromae has been previously reported in cashew and is the most prevalent species observed. All the other species are reported here for the first time on this host. All species of Botryosphaeriaceae were pathogenic on detached green cashew shoots. Differences in aggressiveness were observed among the species, with N. batangarum, L. iraniensis, L. jatrophicola, and L. gravistriata characterized as the most aggressive species, whilst L. euphorbicola and L. pseudotheobromae were identified as the least aggressive., (Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Augmented reactive oxygen species levels consequential to functional alteration of key mitochondrial attributes contribute to carcinogenesis, either directly via oxidative DNA damage infliction or indirectly via activation of oncogenic signaling cascades. We previously reported activation of a key oncogenic signaling cascade via mammalian target of rapamycin (mTOR) signaling complex-2 (mTORC2) owing to estrogen receptor (ER-α)-dependent augmentation of O 2 .- within the mitochondria of 17-β-estradiol (E2)-stimulated breast cancer cells. Manganese superoxide dismutase (MnSOD) is the principal mitochondrial attribute governing mitochondrial O 2 .- homeostasis, raising the possibility that its functional alteration could be instrumental in augmenting mitochondrial O 2 .- levels in breast cancer cells. Here we show ER-dependent transient inhibition of MnSOD catalytic function in breast cancer cells. Catalytic function of MnSOD is tightly regulated at the post-translational level. Post-translational modifications such as phosphorylation, nitration and acetylation represent key regulatory means governing the catalytic function of MnSOD. Acetylation at lysine-68 (K68) inhibits MnSOD catalytic activity and thus represents an important post-translational regulatory mechanism in human cells. Using reciprocal immunoprecipitation and proximity ligation assay, we demonstrate the occurrence of direct physical interaction between ER-α and MnSOD in human breast cancer cells, which in turn was associated with potentiated acetylation of MnSOD at K68. In addition, we also observed diminished interaction of MnSOD with sirtuin-3, the key mitochondrial deacetylase that deacetylates MnSOD at critical K68 and thereby activates it for scavenging O 2 .- . Consequently, compromised deacetylation of MnSOD at K68 leading to its inhibition and a resultant buildup of O 2 .- within the mitochondria culminated in the activation of mTORC2. In agreement with this, human breast cancer tissue specimen exhibited a positive correlation between acetyl-MnSOD K68 levels and phospho-Ser 2481 mTOR levels. In addition to exposing the crosstalk of ER-α with MnSOD post-translational regulatory mechanisms, these data also unravel a regulatory role of ER/MnSOD interaction as an important control switch for redox regulation of ER-α-responsive oncogenic signaling cascades. Furthermore, our study provides a mechanistic link for ER-α-dependent O 2 .- potentiation and resultant mTORC2 activation in breast cancer cells.

Interleukin 17A (IL-17A) has been associated with protective rather than pathogenic response in Chagas disease (ChD). However, it is not established whether or not IL-17A-mediated immune response is correlated with patient's left ventricular (LV) function in ChD. To address this question we have gathered cardiac functional parameters from ChD patients and analysed the possible relationship between their plasma IL-17A levels and LV function. Plasma IL-17A levels were measured by BD Cytometric Bead Array (CBA) in 240 patients with positive specific serology for Trypanosoma cruzi (T. cruzi) grouped as indeterminate (IND) and Chagas cardiomyopathy (CARD) forms. The levels of IL-17A in ChD patients were compared with 32 healthy individuals, mean age of 39 years, 50% male, that were also included as a control group (non-infected [NI]). The overall mean age of ChD patients was 46 years and 52% were male. The IND group included 95 asymptomatic patients, with ages ranging from 27 to 69 years (mean of 43 years), and 42.1% of them were male. The CARD group included 145 patients, which 58.6% were male, with ages ranging from 23 to 67 years (mean of 49). The IND group presented substantially higher levels of IL-17A, median of 26.16 (3.66-48.33) as compared to both the CARD group, median of 13.89 (3.87-34.54) (P <0.0001), and the NI group, median of 10.78 (6.23-22.26) (P <0.0001). The data analysis demonstrated that the IND group comprises a significantly greater proportion (P <0.001) of high IL-17A producers (52.6%, 50 of 95 subjects) than do the other groups. A significant direct correlation was verified between IL-17A levels and cardiac function expressed by LV ejection fraction (LVEF), LV diastolic diameter (LVDd), and body surface area (BSA)-indexed LVDd as well as ratio of the early diastolic transmitral flow velocity to early diastolic mitral annular velocity (E/e') in both groups. We demonstrated that plasma IL-17A levels has an accurate sensitivity and specificity to predict heart failure in serology-positive patients and might be a useful parameter to distinguish patients with or without cardiac impairment. This study indicates a consistent relationship between high expression of IL-17A and better LV in human chronic ChD. Our data raise the possibility that IL-17A plays an important immunomodulatory role in the chronic phase of ChD and might be involved in protection against myocardial damage.

Involuntary choreiform movements are clinical hallmark of Huntington's disease, an autosomal dominant neurodegenerative disorder caused by an increased number of CAG trinucleotide repeats in the huntingtin gene. Involuntary movements start with an impairment of facial muscles and then affect trunk and limbs muscles. Huntington's disease symptoms are caused by changes in cortex and striatum neurons induced by mutated huntingtin protein. However, little is known about the impact of this abnormal protein in spinal cord motoneurons that control movement. Therefore, in this study we evaluated abnormalities in the motor unit (spinal cervical motoneurons, motor axons, neuromuscular junctions and muscle) in a mouse model for Huntington's disease (BACHD). Using light, fluorescence, confocal, and electron microscopy, we showed significant changes such as muscle fibers atrophy, fragmentation of neuromuscular junctions, axonal alterations, and motoneurons death in BACHD mice. Noteworthy, the surviving motoneurons from BACHD spinal cords were smaller than WT. We suggest that this loss of larger putative motoneurons is accompanied by a decrease in the expression of fast glycolytic muscle fibers in this model for Huntington's disease. These observations show spinal cord motoneurons loss in BACHD that might help to understand neuromuscular changes in Huntington's disease., (© 2016 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

The acquisition of reliable data strongly depends on experimental design. When studying the effects of light on processes such as behaviour and physiology it is crucial to maintain all environmental conditions constant apart from the one under study. Furthermore, the precise values of the environmental factors applied during the experiment should be known. Although seemingly obvious, these conditions are often not met when the effects of light are being studied. Here, we document and discuss the wavelengths and light intensities of natural and artificial light sources. We present standardised experimental protocols together with building plans of a custom made instrument designed to accurately control light and temperature for experiments using fresh water or marine species. Infrared light is commonly used for recording behaviour and in electrophysiological experiments although the properties of fish photoreceptors potentially allow detection into the far red. As an example of our experimental procedure we have applied our protocol and instrument to specifically test the impact of infrared light (840 nm) on the zebrafish circadian clock, which controls many aspects of behaviour, physiology and metabolism. We demonstrate that infrared light does not influence the zebrafish circadian clock. Our results help to provide a solid framework for the future study of light dependent processes in aquatic organisms.

Adeno-associated virus (AAV) vectors have emerged as a safe and efficient gene therapy platform. One complication is that a significant amount of empty particles have always been generated as impurities during AAV vector production. However, the effects of such particles on AAV vector performance remain unclear. Here we systemically evaluated the biological properties of three types of "empty" AAV particles: syngeneic pseudo-vectors with partial AAV genomes derived from DNA of the corresponding full particles, allogeneic pseudo-vectors with partial genomes different from the corresponding full particles, and null pseudo-vectors with no DNA inside the capsids. The syngeneic particles in excess increased the corresponding full AAV vector transgene expression both in vivo and in vitro. However, such effects were not observed with null or allogeneic particles. The observed differences among these pseudo-AAV particles may be ascribed to the syngeneic pseudo-vector DNA facilitating the complementary DNA synthesis of the corresponding full AAV particles. Our study suggests that the DNA content in the pseudo-vectors plays a key role in dictating their effects on AAV transduction. The effects of residual "empty" particles should be adequately assessed when comparing AAV vector performance. The syngeneic AAV pseudo-vectors may be used to enhance the efficacy of gene therapy.

Recombinant adeno-associated viral (rAAV) vectors have recently achieved clinical successes in human gene therapy. However, the commonly observed, heavier particles found in rAAV preparations have traditionally been ignored due to their reported low in vitro transduction efficiency. In this study, the biological properties of regular and high-density rAAV serotype 8 vectors, rAAV RD and rAAV HD , were systemically compared. Results demonstrated that both rAAV RD and rAAV HD exhibited similar DNA packaging profiles, while rAAV HD capsids contained fewer VP1 and VP2 proteins, indicating that the rAAV HD particles contained a higher DNA/protein ratio than that of rAAV RD particles. Dynamic light scattering and transmission electron microscopy data revealed that the diameter of rAAV HD was smaller than that of rAAV RD . In vitro, rAAV HD was two- to fourfold less efficient in transduction compared with rAAV RD . However, the transduction performance of rAAV HD and rAAV RD was similar in vivo. No significant difference in neutralizing antibody formation against rAAV RD and rAAV HD was observed, suggesting that the surface epitopes of rAAV RD and rAAV HD are congruent. In summary, the results of this study demonstrate that rAAV RD and rAAV HD are equally competent for in vivo transduction, despite their difference in vitro. Therefore, the use of rAAV HD vectors in human gene therapy should be further evaluated., Competing Interests: Author Disclosure No competing financial interests exist.

A series of novel 2-(chromon-3-yl)-4,5-diphenyl-1 H -imidazoles ( 4a - h ) were synthesized by one pot condensation of substituted 3-formylchromones ( 1a - h ), benzil (2) and ammonium acetate (3) in refluxing acetic acid at 110 °C under N2 atmosphere. Allylation of compounds 4a - h with allyl bromide in the presence of fused K 2 CO 3 furnished N- allyl - 2-(chromon-3-yl)-4,5-diphenyl-1 H -imidazoles ( 6a - h ). The synthesized compounds were characterized spectroscopically and evaluated for in vitro antimicrobial activity against various pathogenic bacterial and fungal strains by disc diffusion method. Compounds bearing electron withdrawing substituents such as bromo ( 4f ) showed significant inhibitory activity against S . cerevisiae (MIC 1.4 μg/ml) and 4g containing chloro substituent, displayed more inhibitory potential against C . albicans (MIC 1.5), as compared to the standard drugs. Compounds 6a and 4c exhibit remarkable inhibitory potential against B . subtilis with MIC 0.98 and 1.23, respectively. The time kill assay for active compound 6a was performed by viable cell count (VCC) method to elucidate the microbicidal nature of 2-(chromon-3-yl)imidazoles. A molecular docking study of most active compounds with target 'lanosterol 14α-demethylase' (CYP51) was performed to unravel the mode of antifungal action.

Transient, multi-protein complexes are important facilitators of cellular functions. This includes the chaperome, an abundant protein family comprising chaperones, co-chaperones, adaptors, and folding enzymes-dynamic complexes of which regulate cellular homeostasis together with the protein degradation machinery. Numerous studies have addressed the role of chaperome members in isolation, yet little is known about their relationships regarding how they interact and function together in malignancy. As function is probably highly dependent on endogenous conditions found in native tumours, chaperomes have resisted investigation, mainly due to the limitations of methods needed to disrupt or engineer the cellular environment to facilitate analysis. Such limitations have led to a bottleneck in our understanding of chaperome-related disease biology and in the development of chaperome-targeted cancer treatment. Here we examined the chaperome complexes in a large set of tumour specimens. The methods used maintained the endogenous native state of tumours and we exploited this to investigate the molecular characteristics and composition of the chaperome in cancer, the molecular factors that drive chaperome networks to crosstalk in tumours, the distinguishing factors of the chaperome in tumours sensitive to pharmacologic inhibition, and the characteristics of tumours that may benefit from chaperome therapy. We find that under conditions of stress, such as malignant transformation fuelled by MYC, the chaperome becomes biochemically 'rewired' to form a network of stable, survival-facilitating, high-molecular-weight complexes. The chaperones heat shock protein 90 (HSP90) and heat shock cognate protein 70 (HSC70) are nucleating sites for these physically and functionally integrated complexes. The results indicate that these tightly integrated chaperome units, here termed the epichaperome, can function as a network to enhance cellular survival, irrespective of tissue of origin or genetic background. The epichaperome, present in over half of all cancers tested, has implications for diagnostics and also provides potential vulnerability as a target for drug intervention., Competing Interests: The authors declare competing financial interests: details are available in the online version of the paper. Readers are welcome to comment on the online version of the paper

Background: Although type 1 diabetes (T1D) remains the most frequent form of diabetes in individuals aged less than 20 years at onset, other forms of diabetes are being increasingly recognized., Objectives: To describe the population of children with other forms of diabetes (non-type 1) included in the multinational SWEET (Better control in Pediatric and Adolescent diabeteS: Working to crEate CEnTers of Reference) database for children with diabetes., Methods: Cases entered in the SWEET database are identified by their physician as T1D, type 2 diabetes (T2D) and other types of diabetes according to the ISPAD classification. Etiologic subgroups are provided for other types of diabetes. Descriptive analyses were tabulated for age at onset, gender, daily insulin doses, and hemoglobin A1c (A1C) for each type and subtype of diabetes and when possible, values were compared., Results: Of the 27 104 patients included in this report, 95.5% have T1D, 1.3% T2D, and 3.2% other forms of diabetes. The two most frequent etiologies for other forms of diabetes were maturity onset diabetes of the young (MODY) (n = 351) and cystic fibrosis-related diabetes (CFRD) (n = 193). The cause was unknown or unreported in 10% of other forms of diabetes. Compared with T1D, children with T2D and CFRD were diagnosed at an older age, took less insulin and had lower A1C (all P < .0001)., Conclusion: In centers included in SWEET, forms of diabetes other than type 1 remain rare and at times difficult to characterize. Sharing clinical information and outcome between SWEET centers on those rare forms of diabetes has the potential to improve management and outcome., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

In many commercially available and in-house-prepared reference electrodes, nanoporous glass frits (often of the brand named Vycor) contain the electrolyte solution that forms a salt bridge between the sample and the reference solution. Recently, we showed that in samples with low ionic strength, the half-cell potentials of reference electrodes comprising nanoporous Vycor frits are affected by the sample and can shift in response to the sample composition by more than 50 mV (which can cause up to 900% error in potentiometric measurements). It was confirmed that the large potential variations result from electrostatic screening of ion transfer through the frit due to the negatively charged surfaces of the glass nanopores. Since the commercial production of porous Vycor glass was recently discontinued, new materials have been used lately as porous frits in commercially available reference electrodes, namely frits made of Teflon, polyethylene, or one of two porous glasses sold under the brand names CoralPor and Electro-porous KT. In this work, we studied the effect of the frit characteristics on the performance of reference electrodes, and show that the unwanted changes in the reference potential are not unique to electrodes with Vycor frits. Increasing the pore size in the glass frits from the <10 nm into the 1 μm range or switching to polymeric frits with pores in the 1 to 10 μm range nearly eliminates the potential variations caused by electrostatic screening of ion transport through the frit pores. Unfortunately, bigger frit pores result in larger flow rates of the reference solution through the pores, which can result in the contamination of test solutions.

Background: Immunological characterization of mycobacterial peptides may help not only in the preparation of a vaccine for leprosy but also in developing in vitro T-cell assays that could perhaps be used as an in vitro correlate for treatment outcome. The main goal of this study was to evaluate the use of Mycobacterium bovis recombinant 32-kDa protein (r32-kDa) antigen-stimulated T-cell assay as a surrogate marker for treatment outcome and monitor vitamin D receptor (VDR)-mediated anti-microbial responses during multidrug therapy (MDT) in leprosy., Methods: Newly diagnosed tuberculoid and lepromatous leprosy patients were enrolled and followed up during their course of MDT at 6 and 12 months. IFN-γ, IL-10, IL-17 and IL-23 levels in culture supernatants and expression of VDR, TLR2, LL37 and DEFB in r32-kDa-stimulated PBMCs were measured. Controls comprised household contacts (HHCs) and healthy endemic subjects (HCs)., Results: Significant differences were observed in the levels of IFN-γ, IL-17, IL-23, VDR and anti-microbial peptides LL37 and DEFB after treatment and when compared with that of HHCs and HCs, respectively., Conclusions: These findings suggest that responses to r32-kDa antigen reflect an improved immunological and anti-microbial response in leprosy patients during therapy, thereby indicating its potential use as an immune correlate in the treatment of leprosy patients., (© The Japanese Society for Immunology. 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

This study was carried out to ascertain the allelic frequency of α(+)-thalassemia (α(+)-thal) in Scheduled caste and scheduled tribe populations of the Damoh district of Madhya Pradesh, India. Random blood samples of Scheduled tribe (267) and Scheduled caste (168), considering the family as a sampling unit, were analyzed for the presence of the -α(3.7) (rightward) (NG&#95;000006.1: g.34164&#95;37967del3804) and -α(4.2) (leftward) (AF221717) deletions. α(+)-Thal was significantly higher in the Scheduled tribals (77.9%) as compared to the scheduled caste population (9.0%). About 58.0% scheduled tribals carried at least one chromosome with the -α(3.7) deletion and 20.0% scheduled tribals carried the -α(4.2) deletion. Frequency for the -α(3.7) allele was 0.487 in the scheduled tribal populations in comparison to 0.021 in scheduled castes. Allelic frequency for -α(4.2) was 0.103 and 0.024, respectively, in the above communities. No Hardy-Weinberg equilibrium for α-thal gene (p < 0.05) was detected in the tribal population, indicating the presence of selection pressures in favor of α-thal mutation and adaptation.

The canine is the most important large animal model for testing novel hemophilia A (HA) treatment. It is often necessary to use canine factor VIII (cFIII) gene or protein for the evaluation of HA treatment in the canine model. However, different biological properties between cFVIII and human FVIII (hFVIII) indicated that the development of novel HA treatment may require careful characterization of non-human FVIII. To investigate whether the data obtained using cFVIII can translate to HA treatment in human, we analyzed the differential biological properties of canine heavy chain (cHC) and light chain (cLC) by comparing with human heavy chain (hHC) and light chain (hLC). The secretion of cHC was 5-30-fold higher than hHC, with or without light chains (LCs). cHC+hLC group exhibited ~18-fold increase in coagulation activity compared with hHC+hLC delivery by recombinant adeno-associated viral vectors. Unlike hHC, the secretion of cHC was independent of LCs. cLC improves the specific activity of FVIII by two- to threefold compared with hLC. Moreover, the cLC, but not cHC, contributes to the higher stability of cFVIII. Our results suggested that the cFVIII expression results in the canine model should be interpreted with caution as the cHC secreted more efficiently than hHC and cLC exhibited a more active and stable phenotype than hLC.

The utilization of several sugars in Escherichia coli is regulated by the Phosphotransferase System (PTS), in which diverse sugar utilization modules compete for phosphoryl flux from the general PTS proteins. Existing theoretical work predicts a winner-take-all outcome when this flux limits carbon uptake. To date, no experimental work has interrogated competing PTS uptake modules with single-cell resolution. Using time-lapse microscopy in perfused microchannels, we analyzed the competition between N-acetyl-glucosamine and sorbitol, as representative PTS sugars, by measuring both the expression of their utilization systems and the concomitant impact of sugar utilization on growth rates. We find two distinct regimes: hierarchical usage of the carbohydrates, and co-expression of the genes for both systems. Simulations of a mathematical model incorporating asymmetric sugar quality reproduce our metabolic phase diagram, indicating that under conditions of nonlimiting phosphate flux, co-expression is due to uncoupling of both sugar utilization systems. Our model reproduces hierarchical winner-take-all behaviour and stochastic co-expression, and predicts the switching between both strategies as a function of available phosphate flux. Hence, experiments and theory both suggest that PTS sugar utilization involves not only switching between the sugars utilized but also switching of utilization strategies to accommodate prevailing environmental conditions., (© 2016 John Wiley & Sons Ltd.)

Introduction: The stem of dioecious Tinospora cordifolia (Menispermaceae) is a commonly used traditional Ayurvedic medicine in India having several therapeutic properties., Objective: To develop and validate LC-MS methods for the identification and simultaneous quantitation of various secondary metabolites and to study metabolomic variations in the stem of male and female plants., Methods: Ethanolic extract of stems were analysed by HPLC/ESI-QTOF-MS/MS for rapid screening of bioactive phytochemicals. High resolution MS and MS/MS in positive ESI mode were used for structural investigation of secondary metabolites. An UPLC/ESI-QqQ(LIT) -MS/MS method in MRM mode was developed and validated for the simultaneous quantitation of five bioactive alkaloids., Results: Identification and characterisation of 36 metabolites including alkaloids, sesquiterpenes and phytoecdysteroids were performed using LC-MS and MS/MS techniques. The bioactive alkaloids such as jatrorrhizine, magnoflorine, isocorydine, palmatine and tetrahydropalmatine were successfully quantified in male and female plants. The mean abundances of magnoflorine jatrorrhizine, and oblongine were significantly (P < 0.05) higher in male plants while mean abundances of tetrahydropalmatine, norcoclaurine, and reticuline were significantly (P < 0.05) higher in female plants., Conclusions: Phytochemicals in the stem of male and female Tinospora cordifolia showed significant qualitative and quantitative variations. LC-MS and MS/MS methods can be used to differentiate between male and female plants based on their chemical profiles and quantities of the marker bioactive alkaloids. This chemical composition difference was also evident during vegetative stage when there were no male and female flowers., (Copyright © 2015 John Wiley & Sons, Ltd.)

Key parameters that influence the specific energy of electrochemical double-layer capacitors (EDLCs) are the double-layer capacitance and the operating potential of the cell. The operating potential of the cell is generally limited by the electrochemical window of the electrolyte solution, that is, the range of applied voltages within which the electrolyte or solvent is not reduced or oxidized. Ionic liquids are of interest as electrolytes for EDLCs because they offer relatively wide potential windows. Here, we provide a systematic study of the influence of the physical properties of ionic liquid electrolytes on the electrochemical stability and electrochemical performance (double-layer capacitance, specific energy) of EDLCs that employ a mesoporous carbon model electrode with uniform, highly interconnected mesopores (3DOm carbon). Several ionic liquids with structurally diverse anions (tetrafluoroborate, trifluoromethanesulfonate, trifluoromethanesulfonimide) and cations (imidazolium, ammonium, pyridinium, piperidinium, and pyrrolidinium) were investigated. We show that the cation size has a significant effect on the electrolyte viscosity and conductivity, as well as the capacitance of EDLCs. Imidazolium- and pyridinium-based ionic liquids provide the highest cell capacitance, and ammonium-based ionic liquids offer potential windows much larger than imidazolium and pyridinium ionic liquids. Increasing the chain length of the alkyl substituents in 1-alkyl-3-methylimidazolium trifluoromethanesulfonimide does not widen the potential window of the ionic liquid. We identified the ionic liquids that maximize the specific energies of EDLCs through the combined effects of their potential windows and the double-layer capacitance. The highest specific energies are obtained with ionic liquid electrolytes that possess moderate electrochemical stability, small ionic volumes, low viscosity, and hence high conductivity, the best performing ionic liquid tested being 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide.

Regio- and stereoselective 1,3-dipolar cycloadditions of C-(chrom-4-one-3-yl)-N-phenylnitrones (N) with different mono-substituted, disubstituted, and cyclic dipolarophiles were carried out to obtain substituted N-phenyl-3'-(chrom-4-one-3-yl)-isoxazolidines (1-40). All the synthesized compounds were assayed for their in vitro antibacterial activity and display significant inhibitory potential; in particular, compound 32 exhibited good inhibitory activity against Salmonella typhymurium-1 & Salmonella typhymurium-2 with minimum inhibitory concentration value of 1.56 μg/mL and also showed good potential against methicillin-resistant Staphylococcus aureus with minimum inhibitory concentration 3.12 μg/mL. Quantitative structure activity relationship investigations with stepwise multiple linear regression analysis and docking simulation studies have been performed for validation of the observed antibacterial potential of the investigated compounds for determination of the most important parameters regulating antibacterial activities., (© 2015 John Wiley & Sons A/S.)

Aims/introduction: The changes in metabolic parameters in type 2 diabetic patients who fast during Ramadan have not been studied in Singapore. This study aimed to examine the trends of glycated hemoglobin (HbA1c), systolic blood pressure, low-density lipoprotein cholesterol, and triglycerides in diabetic patients with varying degrees of glycemic control and different types of therapeutic approaches during Ramadan., Methods: The present retrospective study used a national electronic database to examine the metabolic parameter of Malay patients with type 2 diabetes. Eligible patients were stratified into three groups based on their mean HbA1c control before Ramadan: group 1 (HbA1c ≥10%), group 2 (HbA1c 7.1-9.9%) and group 3 (HbA1c ≤7%). Patients with a glomerular filtration rate <15 mL/min were excluded. The trends of metabolic parameters were traced before, during and after Ramadan., Results: Of 13,565 patients examined, 5,172 patients (38.1%) were eligible for this study. Mean change of HbA1c varied from -1.4% to +0.2% during Ramadan, with the greatest reduction observed in group 1 (P < 0.001). A minimal systolic blood pressure reduction was observed in groups 2 and 3 (2 mmHg; P < 0.01). Low-density lipoprotein cholesterol and triglycerides changes were insignificant. A small, 0.1%, reduction in mean HbA1c was observed in patients taking oral antidiabetic agents during Ramadan (P < 0.001)., Conclusions: Blood glucose was most affected during Ramadan, particularly in patients with mean baseline HbA1c ≥10%. The type of antidiabetic agent used did not seem to contribute to glycemic changes.

The widespread application of silver in consumer products and the resulting contamination of natural environments with silver raise questions about the toxicity of Ag(+) in the ecosystem. Natural organic matter, NOM, which is abundant in water supplies, soil, and sediments, can form stable complexes with Ag(+), altering its bioavailability and toxicity. Herein, the extent and kinetics of Ag(+) binding to NOM, matrix effects on Ag(+) binding to NOM, and the effect of NOM on Ag(+) toxicity to Shewanella oneidensis MR-1 (assessed by the BacLight viability assay) were quantitatively studied with fluorous-phase Ag(+) ion-selective electrodes (ISEs). Our findings show fast kinetics of Ag(+) and NOM binding, weak Ag(+) binding for Suwannee River humic acid, fulvic acid, and aquatic NOM, and stronger Ag(+) binding for Pony Lake fulvic acid and Pahokee Peat humic acid. We quantified the effects of matrix components and pH on Ag(+) binding to NOM, showing that the extent of binding greatly depends on the environmental conditions. The effect of NOM on the toxicity of Ag(+) does not correlate with the extent of Ag(+) binding to NOM, and other forms of silver, such as Ag(+) reduced by NOM, are critical for understanding the effect of NOM on Ag(+) toxicity. This work also shows that fluorous-phase Ag(+) ISEs are effective tools for studying Ag(+) binding to NOM because they can be used in a time-resolved manner to monitor the activity of Ag(+) in situ with high selectivity and without the need for extensive sample preparation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Single Molecule, Real-Time (SMRT) Sequencing (Pacific Biosciences, Menlo Park, CA, USA) provides the longest continuous DNA sequencing reads currently available. However, the relatively high error rate in the raw read data requires novel analysis methods to deconvolute sequences derived from complex samples. Here, we present a workflow of novel computer algorithms able to reconstruct viral variant genomes present in mixtures with an accuracy of >QV50. This approach relies exclusively on Continuous Long Reads (CLR), which are the raw reads generated during SMRT Sequencing. We successfully implement this workflow for simultaneous sequencing of mixtures containing up to forty different >9 kb HIV-1 full genomes. This was achieved using a single SMRT Cell for each mixture and desktop computing power. This novel approach opens the possibility of solving complex sequencing tasks that currently lack a solution., (© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Metallo-β-lactamases (MBLs) hydrolyze almost all classes of β-lactam antibiotic, including carbapenems-currently first choice drugs for opportunistic infections by Gram-negative bacterial pathogens. MBL inhibitor development is complicated by the diversity within this group of enzymes, and by the appearance of new enzymes that continue to be identified both as chromosomal genes and on mobile genetic elements. One such newly discovered MBL is DIM-1, a mobile enzyme originally discovered in the opportunist pathogen Pseudomonas stutzeri but subsequently identified in other species and locations. DIM-1 is a subclass B1 MBL more closely related to the TMB-1, GIM-1 and IMP enzymes than to other clinically encountered MBLs such as VIM and NDM; and possesses Arg, rather than the more usual Lys, at position 224 in the putative substrate binding site. Here we report the crystallization and structure determination of DIM-1. DIM-1 possesses a binuclear metal center with a 5 (rather than the more usual 4) co-ordinate tri-histidine (Zn1) site and both 4- and 5-co-ordinate Cys-His-Asp- (Zn2) sites observed in the two molecules of the crystallographic asymmetric unit. These data indicate a degree of variability in metal co-ordination geometry in the DIM-1 active site, as well as facilitating inclusion of DIM-1 in structure-based MBL inhibitor discovery programmes.

Background: We studied preferences on continuing or forgoing different types of treatments at the end of life in two groups: the general public and people with an advance directive (AD). Furthermore, we studied factors associated with these preferences and whether people's preferences concurred with the content of their AD., Methods: A representative sample of the Dutch population (n=1402) and a cohort of people who own an AD, consisting of members of Right to Die-NL (NVVE, n=5661) and the Christian-orientated Nederlandse Patiënten Vereniging (NPV, n=1059), answered written questionnaires in 2005 or 2007. We used two hypothetical scenarios, about cancer and dementia, and asked questions about continuing or forgoing four medical treatments., Results: A majority of the Dutch public (62-87%) and NVVE members (88-99%) wanted to forgo the different treatments in both scenarios, while members of the NPV generally wanted to continue treatment (46-73%). In all three groups, in both scenarios, a substantial group (13-38%) had different preferences for the different treatments. People were more explicit in their preferences in case of dementia than in case of cancer. Being female, over 55 years of age, having had a higher education and having no (significant) religion increased the odds to refuse treatment. ADs that gave the direction to refuse treatment generally concurred with the preference of their owners (85-98% wanting to refuse treatments)., Discussion: The fact that people with and without ADs have different preferences concerning different treatments and diseases stresses the importance of communication surrounding decision making at the end of life., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)

The ability to tune polymer monolith porosity on multiple length scales is desirable for applications in liquid separations, catalysis, and bioengineering. To this end, we have developed a facile synthetic route to nanoporous polymer monoliths based on controlled polymerization of styrene and divinylbenzene from a poly(lactide) macro-chain transfer agent in the presence of nonreactive poly(ethylene oxide) (PEO). Simple variations in the volume fraction and/or molar mass of PEO lead to either polymerization-induced microphase separation or simultaneous macro- and microphase separation. These processes dictate the resultant morphology and allow for control of the macro- and microstructure of the monoliths. Subsequent selective etching produces monoliths with morphologies that can be tailored from mesoporous, with control over mesopore size, to hierarchically meso- and macroporous, with percolating macropores. This convenient synthetic route to porous polymer monoliths has the potential to be useful in applications where both rapid mass transport and a high surface area are required.

The colloidal stability of silver nanoparticles (AgNPs) in natural aquatic environments influences their transport and environmental persistence, while their dissolution to Ag(+) influences their toxicity to organisms. Here, we characterize the colloidal stability, dissolution behavior, and toxicity of two industrially relevant classes of AgNPs (i.e., AgNPs stabilized by citrate or polyvinylpyrrolidone) after exposure to natural organic matter (NOM, i.e., Suwannee River Humic and Fulvic Acid Standards and Pony Lake Fulvic Acid Reference). We show that NOM interaction with the nanoparticle surface depends on (i) the NOM's chemical composition, where sulfur- and nitrogen-rich NOM more significantly increases colloidal stability, and (ii) the affinity of the capping agent for the AgNP surface, where nanoparticles with loosely bound capping agents are more effectively stabilized by NOM. Adsorption of NOM is shown to have little effect on AgNP dissolution under most experimental conditions, the exception being when the NOM is rich in sulfur and nitrogen. Similarly, the toxicity of AgNPs to a bacterial model (Shewanella oneidensis MR-1) decreases most significantly in the presence of sulfur- and nitrogen-rich NOM. Our data suggest that the rate of AgNP aggregation and dissolution in aquatic environments containing NOM will depend on the chemical composition of the NOM, and that the toxicity of AgNPs to aquatic microorganisms is controlled primarily by the extent of nanoparticle dissolution.

Background: Vitamin D Receptor (VDR) is a transacting transcription factor which mediates immunomodulatory function and plays a key role in innate and adaptive immune responses through its ligand and polymorphisms in VDR gene may affect its regulatory function., Objective: To investigate the association of three VDR gene polymorphisms (TaqI rs731236, FokI rs2228570 and ApaI rs7975232) with leprosy., Methods: The study group includes 404 participants of which 222 were leprosy patients (paucibacillary=87, multibacillary=135) and 182 healthy controls. Genotyping was done using PCR-RFLP technique. Statistical analysis was performed using SNP Stats and PLINK software., Results: The VDR FokI (rs2228570) ff genotype, ApaI (rs7975232) AA, Aa genotype and haplotype T-f-a, T-F-A were positively associated with leprosy when compared to healthy controls., Conclusion: The two variants at Fok and Apa positions in VDR gene are significantly associated with leprosy. Genotypes at FokI (ff), ApaI (aa) and haplotype (T-F-a, T-f-a) may contribute to the risk of developing leprosy by altering VDR phenotype/levels subsequently modulation of immune response., (Copyright © 2015 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

Theory and climate modelling suggest that the sensitivity of Earth's climate to changes in radiative forcing could depend on the background climate. However, palaeoclimate data have thus far been insufficient to provide a conclusive test of this prediction. Here we present atmospheric carbon dioxide (CO2) reconstructions based on multi-site boron-isotope records from the late Pliocene epoch (3.3 to 2.3 million years ago). We find that Earth's climate sensitivity to CO2-based radiative forcing (Earth system sensitivity) was half as strong during the warm Pliocene as during the cold late Pleistocene epoch (0.8 to 0.01 million years ago). We attribute this difference to the radiative impacts of continental ice-volume changes (the ice-albedo feedback) during the late Pleistocene, because equilibrium climate sensitivity is identical for the two intervals when we account for such impacts using sea-level reconstructions. We conclude that, on a global scale, no unexpected climate feedbacks operated during the warm Pliocene, and that predictions of equilibrium climate sensitivity (excluding long-term ice-albedo feedbacks) for our Pliocene-like future (with CO2 levels up to maximum Pliocene levels of 450 parts per million) are well described by the currently accepted range of an increase of 1.5 K to 4.5 K per doubling of CO2.

Palygorskite-montmorillonite (PM) was studied as a potential sewage treatment effluent filter material for carbamazepine. Batch sorption experiments were conducted as a function of granule size (0.3-0.6, 1.7-2.0 and 2.8mm) and different sewage effluent conditions (pH, ionic strength and temperature). Results showed PM had a mix of fibrous and plate-like morphologies. Sorption and desorption isotherms were fitted to the Freundlich model. Sorption is granule size-dependent and the medium granule size would be an appropriate size for optimizing both flow and carbamazepine retention. Highest and lowest sorption capacities corresponded to the smallest and the largest granule sizes, respectively. The lowest and the highest equilibrium aqueous (Ce) and sorbed (qe) carbamazepine concentrations were 0.4 mg L(-1) and 4.5 mg L(-1), and 0.6 mg kg(-1) and 411.8 mg kg(-1), respectively. Observed higher relative sorption at elevated concentrations with a Freundlich exponent greater than one, indicated cooperative sorption. The sorption-desorption hysteresis (isotherm non-singularity) indicated irreversible sorption. Higher sorption observed at higher rather than at lower ionic strength conditions is likely due to a salting-out effect. Negative free energy and the inverse sorption capacity-temperature relationship indicated the carbamazepine sorption process was favorable or spontaneous. Solution pH had little effect on sorption., (Copyright © 2014 Elsevier B.V. All rights reserved.)

We investigate whether a computational model of V1 can predict how observers rate perceptual differences between paired movie clips of natural scenes. Observers viewed 198 pairs of movies clips, rating how different the two clips appeared to them on a magnitude scale. Sixty-six of the movie pairs were naturalistic and those remaining were low-pass or high-pass spatially filtered versions of those originals. We examined three ways of comparing a movie pair. The Spatial Model compared corresponding frames between each movie pairwise, combining those differences using Minkowski summation. The Temporal Model compared successive frames within each movie, summed those differences for each movie, and then compared the overall differences between the paired movies. The Ordered-Temporal Model combined elements from both models, and yielded the single strongest predictions of observers' ratings. We modeled naturalistic sustained and transient impulse functions and compared frames directly with no temporal filtering. Overall, modeling naturalistic temporal filtering improved the models' performance; in particular, the predictions of the ratings for low-pass spatially filtered movies were much improved by employing a transient impulse function. The correlations between model predictions and observers' ratings rose from 0.507 without temporal filtering to 0.759 (p = 0.01%) when realistic impulses were included. The sustained impulse function and the Spatial Model carried more weight in ratings for normal and high-pass movies, whereas the transient impulse function with the Ordered-Temporal Model was most important for spatially low-pass movies. This is consistent with models in which high spatial frequency channels with sustained responses primarily code for spatial details in movies, while low spatial frequency channels with transient responses code for dynamic events., (© 2015 ARVO.)

This study compared the efficacy and toxicity of Gefitinib, Methotrexate and Methotrexate plus 5-Fluorouracil (5-FU) in patients of recurrent squamous cell carcinoma of head and neck (SCCHN) treated with palliative intent. Patients with recurrent SCCHN not amenable to curative treatment were randomly assigned to Gefitinib, Methotrexate or Methotrexate plus 5-FU arm. The primary end point was overall survival. Secondary end points of interest were objective response rate, toxicity and quality of life. Total 117 patients were analyzed. Median overall survival and objective response rates were 8.8 months, 7.8 months and 8.1 months and 7.7%, 5.0% and 7.9% in Gefitinib, Methotrexate and Methotrexate plus 5-FU arms respectively with no statistically significant difference between 3 arms. Gefitinib had different toxicity profile compared with other arms. Majority of toxicities were Grade 1 or Grade 2. Gefitinib had significant improvement in quality of life during initial months over Methotrexate. There was no suggestion that Gefitinib significantly prolonged overall survival compared with Methotrexate and Methotrexate plus 5-FU. However, improved Quality of Life with manageable toxicities was observed.