Evaluation of the immune response to RTS,S/AS01 and RTS,S/AS02 adjuvanted vaccines: randomized, double-blind study in malaria-naïve adults.
This phase II, randomized, double-blind study evaluated the immunogenicity of RTS,S vaccines containing Adjuvant System AS01 or AS02 as compared with non-adjuvanted RTS,S in healthy, malaria-naïve adults (NCT00443131). Thirty-six subjects were randomized (1:1:1) to receive RTS,S/AS01, RTS,S/AS02, or RTS,S/saline at months 0, 1, and 2. Antibody responses to Plasmodium falciparum circumsporozoite (CS) and hepatitis B surface (HBs) antigens were assessed and cell-mediated immune responses evaluated by flow cytometry using intracellular cytokine staining on peripheral blood mononuclear cells. Anti-CS antibody avidity was also characterized. Safety and reactogenicity after each vaccine dose were monitored. One month after the third vaccine dose, RTS,S/AS01 (160.3 EU/mL [95%CI: 114.1-225.4]) and RTS,S/AS02 (77.4 EU/mL (95%CI: 47.3-126.7)) recipients had significantly higher anti-CS antibody geometric mean titers (GMTs) than recipients of RTS,S/saline (12.2 EU/mL (95%CI: 4.8-30.7); P < 0.0001 and P = 0.0011, respectively). The anti-CS antibody GMT was significantly higher with RTS,S/AS01 than with RTS,S/AS02 (P = 0.0135). Anti-CS antibody avidity was in the same range in all groups. CS- and HBs-specific CD4(+) T cell responses were greater for both RTS,S/AS groups than for the RTS,S/saline group. Reactogenicity was in general higher for RTS,S/AS compared with RTS,S/saline. Most grade 3 solicited adverse events (AEs) were of short duration and grade 3 solicited general AEs were infrequent in the 3 groups. No serious adverse events were reported. In conclusion, in comparison with non-adjuvanted RTS,S, both RTS,S/AS vaccines exhibited better CS-specific immune responses. The anti-CS antibody response was significantly higher with RTS,S/AS01 than with RTS,S/AS02. The adjuvanted vaccines had acceptable safety profiles.


DNA sequencing by microchip electrophoresis using mixtures of high- and low-molar mass poly(N,N-dimethylacrylamide) matrices.
Previous studies have reported that mixed molar mass polymer matrices show enhanced DNA sequencing fragment separation compared with matrices formulated from a single average molar mass. Here, we describe a systematic study to investigate the effects of varying the amounts of two different average molar mass polymers on the DNA sequencing ability of poly(N,N-dimethylacrylamide) (pDMA) sequencing matrices in microfluidic chips. Two polydisperse samples of pDMA, with weight-average molar masses of 3.5 MDa and 770 kDa, were mixed at various fractional concentrations while maintaining the overall polymer concentration at 5% w/v. We show that although the separation of short DNA fragments depends strongly on the overall solution concentration of the polymer, inclusion of the high-molar mass polymer is essential to achieve read lengths of interest (>400 bases) for many sequencing applications. Our results also show that one of the blended matrices, comprised of 3% 3.5 MDa pDMA and 2% 770 kDa pDMA, yields similar sequencing read lengths (>520 bases on average) to the high-molar mass matrix alone, while also providing a fivefold reduction in zero-shear viscosity. These results indicate that the long read lengths achieved in a viscous, high-molar mass polymer matrix are also possible to achieve in a tuned, blended matrix of high- and low-molar mass polymers with a much lower overall solution viscosity.


Doctors' career choices in health systems constrained by national medical exams: A discrete choice experiment.
Previous studies that addressed factors influencing junior doctors' career decisions have failed to consider that the market for medical specialty is regulated and doctors are not always able to pursue their career of choice. There is the concern that the National Medical Exam that doctors have to take in many countries conditions their career choices and is behind the low efficacy of policies for retaining doctors in rural areas. In this paper, we use data from a Discrete Choice Experiment applied to a large sample of Portuguese junior doctors (N = 503) who had already taken their Medical Exam but had not chosen their medical specialty yet. We show that the exam score is de facto a strong predictor for (different) job preferences, but only for doctors who are amongst the highest and the lowest exam scores. For all others-almost half of our sample-geographical attachment (having lived or studied in rural areas or in metropolis) is more relevant in determining specialization decisions. Having a good work-life balance, specially amongst female physicians, is also an important determinant of specialty choice. Using latent-class analysis may help policymakers profiling different groups of doctors and tailoring incentive packages according to their characteristics.


Caring for stoma patients with arthritis and mental incapacities.
Mental incapacities in the form of dementia, Parkinson's disease, Huntington's disease, and the physical incapacities of arthritis are difficult hurdles for an older patient who has to undergo stoma surgery, irrespective of the stoma being temporary or permanent. The loss of intellectual and physical function can cause significant deterioration in the patient's ability to carry out day-to-day activities and can also manifest itself in changes in social behaviour. Caring for these patients in the community will require the nurse to adjust the patient's care accordingly, and provide a holistic and more individualised care package that includes the support of the carer or spouse. This article discusses the ways in which community nurses can help the older stoma patient and carer.


Effects of matrix metalloproteinase (MMP)-2 polymorphisms on responsiveness to antihypertensive therapy of women with hypertensive disorders of pregnancy.
Imbalanced matrix metalloproteinase (MMP) expression, including MMP-2, has been demonstrated in pre-eclampsia. However, little is known about the effect of polymorphisms in MMP-2 gene on hypertensive disorders of pregnancy. We examined whether two functional MMP-2 polymorphisms (g.-1306C>T and g.-735C>T) are associated with pre-eclampsia and/or gestational hypertension and whether these polymorphisms affect therapeutic responses in women with these conditions. We studied 216 healthy pregnant women (HP), 185 patients with gestational hypertension (GH) and 216 patients with pre-eclampsia (PE). They were stratified as responsive or non-responsive to antihypertensive therapy according to clinical and laboratorial parameters of therapeutic responsiveness. Genomic DNA was extracted from whole blood and genotypes for g-1306C>T and g.-735C>T polymorphisms were determined by real-time PCR using Taqman allele discrimination assays. Haplotype frequencies were inferred using the PHASE 2.1 program. The distributions of MMP-2 genotypes and haplotypes were similar in HP, GH and PE patients (p > 0.05). In addition, we found no significant differences in MMP-2 genotype or haplotype frequencies when GH or PE patients were classified as responsive or non-responsive to antihypertensive therapy (p > 0.05). Our results suggest that MMP-2 polymorphisms do not affect the susceptibility to hypertensive disorders of pregnancy. In parallel, MMP-2 polymorphisms apparently do not affect the responsiveness to antihypertensive therapy of women with these hypertensive disorders of pregnancy.


Technical advances of pulmonary endarterectomy for chronic thromboembolic pulmonary hypertension.
Pulmonary endarterectomy is the definitive treatment for chronic pulmonary hypertension as the result of thromboembolic disease. Although significant progress has been made over the last decade in recognition, diagnostic modalities, and treatment of this disease, chronic thromboembolic pulmonary hypertension (CTEPH) continues to be severely underdiagnosed and as a consequence pulmonary endarterectomy remains an uncommon procedure. Patients with CTEPH may present with a variety of debilitating cardiopulmonary symptoms. However, once diagnosed, there is no curative role for medical management, and surgery remains the only option. Medical management in these patients is only palliative, and surgery by means of transplantation for this type of pulmonary hypertension is an inappropriate use of resources with less than satisfactory results. In this article we describe the technical advances of pulmonary endarterectomy and the current procedure as it is performed at University of California-San Diego Medical Center.


Single crystalline ZnO radial homojunction light-emitting diodes fabricated by metalorganic chemical vapour deposition.
ZnO radial p-n junction architecture has the potential for forward-leap of light-emitting diode (LED) technology in terms of higher efficacy and economical production. We report on ZnO radial p-n junction-based light emitting diodes prepared by full metalorganic chemical vapour deposition (MOCVD) with hydrogen-assisted p-type doping approach. The p-type ZnO(P) thin films were prepared by MOCVD with the precursors of dimethylzinc, tert-butanol, and tertiarybutylphosphine. Controlling the precursor flow for dopant results in the systematic change of doping concentration, Hall mobility, and electrical conductivity. Moreover, the approach of hydrogen-assisted phosphorous doping in ZnO expands the understanding of doping behaviour in ZnO. Ultraviolet and visible electroluminescence of ZnO radial p-n junction was demonstrated through a combination of position-controlled nano/microwire and crystalline p-type ZnO(P) radial shell growth on the wires. The reported research opens a pathway of realisation of production-compatible ZnO p-n junction LEDs.


First offers as anchors: the role of perspective-taking and negotiator focus.
Three experiments explored the role of first offers, perspective-taking, and negotiator self-focus in determining distributive outcomes in a negotiation. Across 3 experiments, whichever party, the buyer or seller, made the 1st offer obtained a better outcome. In addition, 1st offers were a strong predictor of final settlement prices. However, when the negotiator who did not make a 1st offer focused on information that was inconsistent with the implications of the opponent's 1st offer, the advantageous effect of making the 1st offer was eliminated: Thinking about one's opponent's alternatives to the negotiation (Experiment 1), one's opponent's reservation price (Experiment 2), or one's own target (Experiment 3) all negated the effect of 1st offers on outcomes. These effects occurred for both face-to-face negotiations and E-mail negotiations. Implications for negotiations and perspective-taking are discussed.


Drought Stress and Root-Associated Bacterial Communities.
Root-associated bacterial communities play a vital role in maintaining health of the plant host. These communities exist in complex relationships, where composition and abundance of community members is dependent on a number of factors such as local soil chemistry, plant genotype and phenotype, and perturbations in the surrounding abiotic environment. One common perturbation, drought, has been shown to have drastic effects on bacterial communities, yet little is understood about the underlying causes behind observed shifts in microbial abundance. As drought may affect root bacterial communities both directly by modulating moisture availability, as well as indirectly by altering soil chemistry and plant phenotypes, we provide a synthesis of observed trends in recent studies and discuss possible directions for future research that we hope will provide for more knowledgeable predictions about community responses to future drought events.


Contribution of capillary endothelium to cor pulmonale development in experimental lung emphysema.
The aim of this study was the ultrastructural analysis of the changes in the endothelium of myocardial capillaries and the processes observed in their vicinity in consecutive stages of experimental lung emphysema. Emphysematous changes in the lungs of Wistar rats were induced with a single intraperitoneal infusion of proteolytic enzyme solution, papain, in a dose of 20 mg/kg b.w./0.5 ml PBS. Ultrastructural analysis of the changes within the right ventricle and the subendocardial part of the left ventricle of the heart was made by a transmission electron microscope after 1, 3 and 6 months following papain administration. Special attention was paid to the analysis of changes found in the vicinity of newly formed myocardial capillaries. Animals killed after 1 and 3 months following papain infusion showed blurred borderline between the endothelium of the newly formed myocardial capillaries and perivascular connective matrix. The new vessels had poorly developed basement membrane. Fibrillary structures and/or collagen fibrils were found instead. The latter frequently demonstrated features of disorders in the structure and spatial distribution. Neither fibroblasts nor their processes were observed in the vicinity of the new collagen fibrils. Pericytes were occasionally present. In the later period (6 months from intratracheal papain infusion) well formed collagen fibres dominated in the vicinity of myocardial capillaries. The results of the ultrastructural analysis suggest the possibility of active contribution of endothelial cells to the processes of perivascular fibrosis within the myocardium in the course of experimental lung emphysema.


Nanoparticles as protein and gene carriers to mucosal surfaces.
One of the most exciting and challenging applications of nanotechnology in medicine is the development of nanocarriers for the intraepithelial delivery of biomacromolecules through mucosal surfaces. These biomacromolecules represent an increasingly important segment of the therapeutic arsenal; however, their potential is still limited by their instability and inability to cross biological barriers. Nanoparticle carriers have emerged as one of the most promising technologies to overcome this limitation, owing mainly to their demonstrated capacity to interact with biological barriers. In this review, we summarize the current advances made on nanoparticles designed for transmucosal delivery. Supported by the examples of a variety of therapeutic macromolecules - peptides and proteins, gene medicines and vaccines - we review the lessons learned from the past and we offer a future perspective for this field.


EEG microstate dynamics indicate a U-shaped path to propofol-induced loss of consciousness.
Evidence suggests that the stream of consciousness is parsed into transient brain states manifesting themselves as discrete spatiotemporal patterns of global neuronal activity. Electroencephalographical (EEG) microstates are proposed as the neurophysiological correlates of these transiently stable brain states that last for fractions of seconds. To further understand the link between EEG microstate dynamics and consciousness, we continuously recorded high-density EEG in 23 surgical patients from their awake state to unconsciousness, induced by step-wise increasing concentrations of the intravenous anesthetic propofol. Besides the conventional parameters of microstate dynamics, we introduce a new implementation of a method to estimate the complexity of microstate sequences. The brain activity under the surgical anesthesia showed a decreased sequence complexity of the stereotypical microstates, which became sparser and longer-lasting. However, we observed an initial increase in microstates' temporal dynamics and complexity with increasing depth of sedation leading to a distinctive "U-shape" that may be linked to the paradoxical excitation induced by moderate levels of propofol. Our results support the idea that the brain is in a metastable state under normal conditions, balancing between order and chaos in order to flexibly switch from one state to another. The temporal dynamics of EEG microstates indicate changes of this critical balance between stability and transition that lead to altered states of consciousness.


Heteroplasmy for the Cytochrome b Gene in Podosphaera xanthii and its Role in Resistance to QoI Fungicides in Spain.
In Spain, management of the cucurbit powdery mildew pathogen Podosphaera xanthii is strongly dependent on chemicals such as quinone outside inhibitor (QoI) fungicides. In a previous report, widespread resistance to QoI fungicides in populations of P. xanthii in south-central Spain was documented, but the molecular mechanisms of resistance remained unclear. In this work, the role of the Rieske-FeS (risp) and the cytochrome b (cytb) gene mutations in QoI resistance of P. xanthii were examined. No point mutations in the risp gene were found in the three QoI-resistant isolates analyzed. For cytb, sequence analysis revealed the presence of a G143A substitution that occurs in many QoI-resistant fungi. This mutation was always detected in QoI-resistant isolates of P. xanthii; however, it was also detected in sensitive isolates. To better understand the role of heteroplasmy for cytb in QoI resistance of P. xanthii, an allele-specific quantitative PCR was developed to quantify the relative abundance of the G143 (sensitive) and A143 (resistant) alleles. High relative abundance of A143 allele (70%) was associated with isolates resistant to QoI fungicides; however, QoI-sensitive isolates also carried the mutated allele in frequencies ranged from 10 to 60%. Our data suggest that G143A mutation in cytb is the primary factor involved in QoI resistance of P. xanthii but the proportion of G143 and A143 alleles in an isolate may determine its QoI resistance level.


Methods for TALEN Evaluation, Use, and Mutation Detection in the Mosquito Aedes aegypti.
The generation and study of transgenic Aedes aegypti mosquitoes provides an essential tool for elucidating the complex molecular biology of this important vector. Within the field, genetic manipulation has surpassed the proof of principle stage and is now utilized in both applied and theoretical vector control strategies. The application of new instruments, technologies and techniques allows ever more controlled experiments to be conducted. In this text we describe microinjection of Ae. aegypti embryos in the context of evaluating and performing genomic editing with transcription activator-like effector nucleases (TALENs).


Expression of human alpha-fetoprotein in yeast.
Human alpha-fetoprotein (AFP) was expressed in Saccharomyces cerevisiae, with a plasmid containing the cDNA sequence for human AFP fused with the rat AFP signal peptide. The recombinant AFP was purified from the yeast lysate by DEAE-cellulose and immunoaffinity chromatography. The amino acid composition and the molecular weight of the recombinant AFP were similar to those of hepatoma AFP. N-terminal amino acids sequence analysis indicated that the signal peptide had been processed. The recombinant and hepatoma AFP reacted identically in Ouchterlony immunodiffusion and radioimmunoassay tests. These observations indicated that the yeast recombinant protein had the properties of native AFP.


Effects of VM26 (teniposide), a specific inhibitor of type II DNA topoisomerase, on SV40 DNA replication in vivo.
Simian Virus 40 (SV40) infected cells were pulse labeled with (3H) thymidine and chased either in the absence or in the presence of the cytotoxic drug VM26 (teniposide). We investigated the structure of labeled SV40 DNA and found that VM26 had no significant effect on replicative chain elongation but strongly inhibited the conversion of late replication intermediates to mature DNA daughter molecules. The late replicative SV40 DNA intermediates which accumulate in VM26 treated cells contained essentially full length labeled DNA strands. These newly synthesized strands were not part of two catenated interlocked SV40 monomers suggesting that the block occurred prior to the final ligation reaction. Since VM26 is known to be a specific inhibitor of DNA topoisomerase II we conclude that this enzyme is dispensable for the chain elongation of replicating SV40 DNA, but that it is essential for the termination of SV40 DNA replication cycles.


Some reasons for disagreement among scorers of infant intelligence test items.
Six videotapes covering 1406 administrations of items from the Griffiths Scales of Mental Development were made, and each one was shown to a different panel of 9 or 10 observers. Each person independently scored the items and made comments on administration and scoring. Reasons for disagreements among scorers were considered in terms of these comments and the characteristics of individuals and groups participating. Disagreement was particularly high for items from the Baby Scales. This was at least partly due to use of mothers' reports. Mothers' reports were found to present a particular problem when elicited on occasions for which they are not recommended by the manual. Across all ages disagreement was related to difficulties with both administrative and scoring criteria. The present study was designed to be sensitive to a range of sources of unreliability and the findings may have relevance for other infant assessment procedures.


Identification of Variable Importance for Predictions of Mortality From COVID-19 Using AI Models for Ontario, Canada.
The Severe Acute Respiratory Syndrome Coronavirus 2 pandemic has challenged medical systems to the brink of collapse around the globe. In this paper, logistic regression and three other artificial intelligence models (XGBoost, Artificial Neural Network and Random Forest) are described and used to predict mortality risk of individual patients. The database is based on census data for the designated area and co-morbidities obtained using data from the Ontario Health Data Platform. The dataset consisted of more than 280,000 COVID-19 cases in Ontario for a wide-range of age groups; 0-9, 10-19, 20-29, 30-39, 40-49, 50-59, 60-69, 70-79, 80-89, and 90+. Findings resulting from using logistic regression, XGBoost, Artificial Neural Network and Random Forest, all demonstrate excellent discrimination (area under the curve for all models exceeded 0.948 with the best performance being 0.956 for an XGBoost model). Based on SHapley Additive exPlanations values, the importance of 24 variables are identified, and the findings indicated the highest importance variables are, in order of importance, age, date of test, sex, and presence/absence of chronic dementia. The findings from this study allow the identification of out-patients who are likely to deteriorate into severe cases, allowing medical professionals to make decisions on timely treatments. Furthermore, the methodology and results may be extended to other public health regions.


Continuing professional development for Physician Associates in primary care.
The Physician Associate role is relatively new to the United Kingdom and is currently undergoing a period of significant expansion. This includes an aim of 1000 PAs working in primary care by 2020. The profession has specific continuing professional development requirements which need to be addressed. These requirements can be met through the deployment of some well established pedagogical strategies which are already in use for junior doctors and allied health professionals.


[Preparing of Al2O3/ZrO2 composite dental ceramics through isostatic pressing technology].
To find out how to prepare high-density dental ceramics through isostatic pressing so that sintering shrinkage will be reduced. To prepare Al2O3/ZrO2 composite powder first, then to mold through dry-pressing, and to shape the green-body through isostatic pressing. The green-bodies were sintered at the temperature of 1 400 degrees C and kept at the temperature for different period of time (2 h, 3 h, 4 h). After that, the density and fracture strength were measured and the microstructure observed by scanning electron microscope (SEM). The sample product's density, line-shrinkage, and fracture strength of ceramics was rising with the sintering time lengthened. The sample product kept under the temperature of 1 400 degrees C for 4 hours, the fracture strength was (497.27 +/- 78.45) MPa and glass phase distributed evenly in the ceramics and the grains were integrated owing to the glass phase. The longer the sintering time, the more even the microstructure was. The sintering quality and the efficiency were improved through isostatic pressing.


Small intestinal obstruction due to Bilharzioma: case report.
Schistosomiasis in endemic areas affects all age groups and present with various clinical manifestations. Small intestinal obstruction as a complication of Schistosomiasis was not described in the literature. This is a report of a 10 year old child from an endemic area with an unusual presentation of subacute small intestinal obstruction. Description of the pathological changes in the intestines following infestation with Schistosomiasis and brief presentation of the control modality in this endemic area are given.


Guest Editorial: Controversies in Fractional Flow Reserve.
Fractional flow reserve (FFR) has been identified as the optimal diagnostic tool to identify significant coronary lesion. However, current evidence does not support this role. The optimal diagnostic strategy should give highly sensitive and specific results with lowest cost and accomplishing this task has been made more difficult in the era following the COURAGE trial.


Gastroenterologist-Level Identification of Small-Bowel Diseases and Normal Variants by Capsule Endoscopy Using a Deep-Learning Model.
Capsule endoscopy has revolutionized investigation of the small bowel. However, this technique produces a video that is 8-10 hours long, so analysis is time consuming for gastroenterologists. Deep convolutional neural networks (CNNs) can recognize specific images among a large variety. We aimed to develop a CNN-based algorithm to assist in the evaluation of small bowel capsule endoscopy (SB-CE) images. We collected 113,426,569 images from 6970 patients who had SB-CE at 77 medical centers from July 2016 through July 2018. A CNN-based auxiliary reading model was trained to differentiate abnormal from normal images using 158,235 SB-CE images from 1970 patients. Images were categorized as normal, inflammation, ulcer, polyps, lymphangiectasia, bleeding, vascular disease, protruding lesion, lymphatic follicular hyperplasia, diverticulum, parasite, and other. The model was further validated in 5000 patients (no patient was overlap with the 1970 patients in the training set); the same patients were evaluated by conventional analysis and CNN-based auxiliary analysis by 20 gastroenterologists. If there was agreement in image categorization between the conventional analysis and CNN model, no further evaluation was performed. If there was disagreement between the conventional analysis and CNN model, the gastroenterologists re-evaluated the image to confirm or reject the CNN categorization. In the SB-CE images from the validation set, 4206 abnormalities in 3280 patients were identified after final consensus evaluation. The CNN-based auxiliary model identified abnormalities with 99.88% sensitivity in the per-patient analysis (95% CI, 99.67-99.96) and 99.90% sensitivity in the per-lesion analysis (95% CI, 99.74-99.97). Conventional reading by the gastroenterologists identified abnormalities with 74.57% sensitivity (95% CI, 73.05-76.03) in the per-patient analysis and 76.89% in the per-lesion analysis (95% CI, 75.58-78.15). The mean reading time per patient was 96.6 ± 22.53 minutes by conventional reading and 5.9 ± 2.23 minutes by CNN-based auxiliary reading (P < .001). We validated the ability of a CNN-based algorithm to identify abnormalities in SB-CE images. The CNN-based auxiliary model identified abnormalities with higher levels of sensitivity and significantly shorter reading times than conventional analysis by gastroenterologists. This algorithm provides an important tool to help gastroenterologists analyze SB-CE images more efficiently and more accurately.


Prevalence and Factors Associated with Current Cigarette Smoking among Ethiopian University Students: A Systematic Review and Meta-Analysis.
Although tobacco use is highest in high-income countries, most tobacco-related deaths occur in low- and middle-income countries with the highest number of deaths recorded in East Africa. The aim of this systematic review and meta-analysis is to determine the pooled prevalence and associated factors of current cigarette smoking among Ethiopian university students. The authors searched databases from PubMed, PsycINFO, Google Scholar, EMBASE, and Web of Sciences. The publications included in the analysis were inclusive, the literature was searched from January 2011 to December 2018. The JBI-MAStARI critical appraisal tool was applied to 13 publications identified by the database search. I2 statistics were used to indicate heterogeneity. Publication bias was evaluated using the visual funnel plot. A p value < 0.1 was considered as indicative of statistically significant publication bias. A random effect meta-analysis model was computed to estimate the pooled prevalence of cigarette smoking, and the variables associated with cigarette smoking were examined. The meta-analysis of 13 studies showed that the pooled prevalence of current cigarette smoking was found to be 12.55% (95% CI: 10.39-14.72; I2 = 94.0%) with no publication bias according to Egger's test (p = 0.007) for cigarette smoking by 2.05 (95% CI: 1.52-2.75). Factors associated with cigarette smoking were peer influence 2.79 (95% CI: 1.62-4.82; I2 = 35.7), khat chewing (95% CI: 2.81-15.26; I2 = 82.5), and alcohol use 11.16 (95% CI: 7.46-16.71). Our findings indicate a high prevalence of cigarette smoking among Ethiopian university students as compared to the general population. Gender, peer influence, khat chewing, and current alcohol use were significantly associated with cigarette smoking. The authors recommend promoting antismoking campaigns, emphasizing health hazard public service announcements about cigarettes, and integrating health education on smoking in youth-friendly services, especially targeting higher educational institutions.


Distinct developmental regulation and properties of the responsiveness of different genes to cyclic AMP in Dictyostelium discoideum.
Cyclic AMP (cAMP) is known to be an important mediator of gene expression in eukaryotic cells. At present, little is known about the developmental events which render specific genes responsive to cAMP in distinct cell types, or about the biochemical mechanisms by which cAMP exerts these regulatory effects. By examining the effects of cAMP treatment on specific mRNA levels in Dictyostelium discoideum cells with different 'developmental histories', we defined the developmental states in which specific genes display responsiveness to cAMP. We focused on two specific rapid responses: the ability of cAMP to inhibit the expression of an 'early' developmentally regulated mRNA (discoidin-I) and to stimulate the expression of a 'late', prespore-specific mRNA (PL3). Using this approach, we showed that, for both mRNAs, the ability to respond rapidly to cAMP is absent from vegetative cells grown on bacteria, and is acquired during development on filters. Furthermore, we identified several developmental states in which the discoidin-I response to cAMP is present, but in which the PL3 response is not. In experiments designed to examine the effects of cAMP analogues on the levels of these two mRNAs, we demonstrated that the analogue specificities of the discoidin-I and PL3 responses are different, and that the specificity for the PL3 response depends on the developmental state. The developmental kinetics and analogue specificity of the PL3 response suggest a two-step mode of action of cAMP in activating the expression of this gene. We discuss possible implications of these findings for the mechanisms of action of exogenous cAMP as well as for the role of cAMP in controlling the changes in gene expression that accompany normal development.


Expression of oncogenes in human tumours with special reference to the head and neck region.
A major recent advance in cancer research has been in the field of oncogenes. Oncogenes are genes with a proven cancer association and which appear to be particularly implicated in cellular regulation and proliferation. The oncogenic potential of specific cellular genes has now been recognised and this has influenced current thinking concerning the initiation of carcinogenesis. The specific role of an oncogene is still incompletely understood but research with one particular oncogene (ras) has demonstrated that it can be involved in more than one stage of multi-step carcinogenesis. New techniques are being developed and evaluated to determine the expression of specific oncogenes in normal and neoplastic tissues, with a view to using them in future diagnostic immuno-histopathological methods. This review describes the concept of oncogenes and discusses their role in the development of neoplasia. The results of the expression of various oncogenes in human malignancies with special reference to the head and neck regions are discussed. Finally, the future prospects of this research field are examined their and its possible implications in cancer therapy.


Deep learning adapted acceleration for limited-view photoacoustic image reconstruction.
The limited-view issue can cause a low-quality image in photoacoustic (PA) computed tomography due to the limitation of geometric condition. The model-based method is used to resolve this problem, which contains different regularization. To adapt fast and high-quality reconstruction of limited-view PA data, in this Letter, a model-based method that combines the mathematical variational model with deep learning is proposed to speed up and regularize the unrolled procedure of reconstruction. A deep neural network is designed to adapt the step of the gradient updated term of data consistency in the gradient descent procedure, which can obtain a high-quality PA image with only a few iterations. A comparison of different model-based methods shows that our proposed scheme has superior performances (over 0.05 for SSIM) with the same iteration (three times) steps. Finally, we find that our method obtains superior results (0.94 value of SSIM for in vivo) with a high robustness and accelerated reconstruction.


[The opinion of patients treated with intracavernous injections: the EASY survey].
The primary objective of the EASY survey (Evaluation de l'Acceptation deS injections dans la dYsfonction érectile) was to study the feelings of patients treated for erectile disorders with long-term intracavernous injections of prostaglandin E1. The major criteria for abandoning treatment with IPDE 5 are, in descending order, inefficacy, cost and poor tolerance. Regarding the results with intracavernous injections of Edex, around 80% of patients were satisfied; 70 to 80% of those treated for risk factors and/or cardiovascular or neurological diseases and 55% of those treated following cancer of the prostate claimed that their sex life had improved.


[Congenital glaucoma].
ōme statistical results have been clarified through research on the infantile glaucoma over the past fifteen years. The incidence of primary glaucoma and secondary, glaucoma was at a ratio of 7:3. Buphthalmos occurred in males and females at a ratio of 2:1. Bilateral and unilateral glaucoma was also seen at a ratio of 2:1. Seventy-five% of the patients visited the clinic within one year of birth. Secondary glaucoma occurred similarly in males and females. The incidence of bilateral and unilateral glaucoma was also similar. Of 14 patients with Sturge-Weber's syndrome, 5 also suffered from Klippel-Trenaunay's syndrome. In addition to these facts, one case with specific Peter's anomary showed evidence of chromosomal abnormality.


pH-Elicited Luminescence Functionalities of Carbon Dots: Mechanistic Insights.
Remarkable and systematic pH-dependent changes are observed in the absorption and emission spectra of carbon dots derived for the first time from lemon juice, a natural bioresource. Detailed photophysical studies of these novel carbon dots (henceforth termed LD), in conjunction with Fourier transform infrared spectra, reveal that among the two possible prototropic equilibria, phenol ↔ phenolate and carboxylic ↔ carboxylate, that occur at the surface of LD, it is the former that is actually coupled with the emissive moiety and directly involved in determining the nature of the electronic energy levels and the associated optical transitions. Apart from providing valuable mechanistic insights on the photoluminescence (PL) of carbon dots, the pH dependence of LD is also demonstrated to yield variable PL signals and perform elementary Boolean logic operations in response to chemical stimulants. The pH effect can therefore complement the optoelectronic functionalities of these promising luminescent nanomaterials and help in the future development of molecular devices and intelligent multianalyte detection systems.


Pharmacogenetics of drug-metabolizing enzymes in Italian populations.
Drug-metabolizing enzymes play a major role in the biotransformation and subsequent elimination of most drugs and xenobiotics from the body. Both phase I and phase II enzymes are highly polymorphic. Inter-individual differences in genes coding for drug-metabolizing enzymes are important for understanding variability in drug response and for individualization of drug prescription. The prevalence of genetic polymorphisms in drug metabolism varies widely with ethnicity, and marked differences in the distribution of allelic variants of genes encoding drug-metabolizing enzymes have been documented in populations of different racial origin. This review aimed to summarize the available studies on genetic polymorphisms associated with drug metabolism conducted in Italian populations and to compare the frequency of the various metabolizer phenotypes and most common variant alleles (and resulting genotypes) with corresponding values from other populations. Notably, published data are not extensive, and most studies were performed on relatively low numbers of individuals. In general, the frequency of polymorphisms in the cytochrome P450 (CYP) genes as well as in the investigated phase II enzymes in the Italian population was similar to values reported for other Caucasian populations. However, the prevalence of CYP2D6 gene duplication among Italians was found to be very high, confirming the higher frequency of CYP2D6 ultrarapid metabolizers in the Mediterranean area compared to Northern Europe. It is worth noting that a geographic gradient in the flavin-containing monooxygenase 3 polymorphism distribution was also seen, the Italian population showing higher similarity to other Mediterranean populations than to North Europeans.


Evolution of T-cell clonality in a patient with Ph-negative acute lymphocytic leukemia occurring after interferon and imatinib therapy for Ph-positive chronic myeloid leukemia.
The development of Philadelphia chromosome (Ph) negative acute leukemia/myelodysplastic syndrome (MDS) in patients with Ph-positive chronic myeloid leukemia (CML) is very rare. The features of restrictive usage and absence of partial T cell clones have been found in patients with CML. However, the T-cell clonal evolution of Ph-negative malignancies during treatment for CML is still unknown. To investigate the dynamic change of clonal proliferation of T cell receptor (TCR) Valpha and Vbeta subfamilies in one CML patient who developed Ph-negative acute lymphoblastic leukemia (ALL) after interferon and imatinib therapy. The peripheral blood mononuclear cells (PBMC) samples were collected at the 3 time points (diagnosis of Ph-positive chronic phase (CP) CML, developing Ph-negative ALL and post inductive chemotherapy (CT) for Ph-negative ALL, respectively). The CDR3 size of TCR Valpha and Vbeta repertoire were detected by RT-PCR. The PCR products were further analyzed by genescan to identify T cell clonality. The CML patient who achieved complete cytogenetic remission (CCR) after 5 years of IFN-alpha therapy suddenly developed Ph-negative ALL 6 months following switch to imatinib therapy. The expression pattern and clonality of TCR Valpha/Vbeta T cells changed in different disease stages. The restrictive expression of Valpha/Vbeta subfamilies could be found in all three stages, and partial subfamily of T cells showed clonal proliferation. Additionally, there have been obvious differences in Valpha/Vbeta subfamily of T cells between the stages of Ph-positive CML-CP and Ph-negative ALL. The Valpha10 and Vbeta3 T cells evolved from oligoclonality to polyclonality, the Vbeta13 T cells changed from bioclonality to polyclonality, when Ph-negative ALL developed. Restrictive usage and clonal proliferation of different Valpha/Vbeta subfamily T cells between the stages of Ph-positive CP and Ph-negative ALL were detected in one patient. These changes may play a role in Ph- negative leukemogenesis.


Quantitative DNA alterations during 5-azacytidine-induced differential modulation of benzo(a)pyrene carcinogenesis in hamster bronchi.
Sustained release implants (SRI) containing 10% benzo(a)pyrene (BP) were placed endobronchially into outbred and syngeneic (F1D) hamsters. Randomly selected OB and F1D hamsters also received 5-azacytidine (AZC), 5 mg/kg i.p., twice weekly until death (AZC-CONT); two more groups of F1D hamsters were given the same AZC dose either for the first 75 days of SRI implantation (AZC-EARLY) or from 80 days after SRI placement until death (AZC-LATE). OB Hamsters were sacrificed at regular intervals from 62 to 189 days of SRI exposure. F1D Hamsters were sacrificed in groups after 120, 150, 180, and 220 days of SRI exposure. The bronchial mucosa at the SRI site was examined cytologically and histologically, as were the tumors that developed. Mean quantitative total cellular DNA values (QDNA) were measured by image analysis. For both varieties of hamster given AZC, QDNA values were higher in early carcinogenesis (CG) and lower in the late stage of CG than in hamsters that did not get AZC (P less than 0.001). QDNA values were lower in epidermoid than in non-epidermoid cancers (P less than 0.001); for both types of cancer, QDNA was lower in AZC-treated hamsters (P less than 0.01). Cancers induced under the influence of AZC included more epidermoid cancers (P less than 0.01) and were of a higher degree of differentiation (P less than 0.01) than those induced by BP alone, especially when AZC was given early in CG. There was no consistent relationship between QDNA and degree of differentiation in tumors. These differential effects of AZC given early during CG suggest that 1) for epidermoid bronchial CG, some of the molecular alterations involved in hyperploidy and in differentiation occur early in the sequential progression of carcinogenesis; and 2) for both epidermoid and non-epidermoid bronchial CG, the necessary changes must occur in a fixed sequence instead of as random events, until all needed changes have occurred.


Both creatine and its product phosphocreatine reduce oxidative stress and afford neuroprotection in an in vitro Parkinson's model.
Creatine is the substrate for creatine kinase in the synthesis of phosphocreatine (PCr). This energetic system is endowed of antioxidant and neuroprotective properties and plays a pivotal role in brain energy homeostasis. The purpose of this study was to investigate the neuroprotective effect of creatine and PCr against 6-hydroxydopamine (6-OHDA)-induced mitochondrial dysfunction and cell death in rat striatal slices, used as an in vitro Parkinson's model. The possible involvement of the signaling pathway mediated by phosphatidylinositol-3 kinase (PI3K), protein kinase B (Akt), and glycogen synthase kinase-3β (GSK3β) was also evaluated. Exposure of striatal slices to 6-OHDA caused a significant disruption of the cellular homeostasis measured as 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide reduction, lactate dehydrogenase release, and tyrosine hydroxylase levels. 6-OHDA exposure increased the levels of reactive oxygen species and thiobarbituric acid reactive substances production and decreased mitochondrial membrane potential in rat striatal slices. Furthermore, 6-OHDA decreased the phosphorylation of Akt (Serine(473)) and GSK3β (Serine(9)). Coincubation with 6-OHDA and creatine or PCr reduced the effects of 6-OHDA toxicity. The protective effect afforded by creatine or PCr against 6-OHDA-induced toxicity was reversed by the PI3K inhibitor LY294002. In conclusion, creatine and PCr minimize oxidative stress in striatum to afford neuroprotection of dopaminergic neurons.


A comparative study of the bioactivation of nitrosamines to mutagens by various animal species including man.
Dimethylnitrosamine, dipropylnitrosamine, methylethylnitrosamine, nitrosopiperidine and nitrosopyrrolidine were assayed for mutagenicity in the Ames test in the presence of hepatic postmitochondrial preparations isolated from the mouse, rat, hamster, pig and man. Prior to each mutagenicity assay all activation systems were fully characterised with respect to monoamine oxidase, mixed-function amine oxidase and mixed-function oxidase activities. The hamster was the most efficient activator for all nitrosamines followed by the mouse. The latter species, however, activated nitrosopyrrolidine only weakly which was the only carcinogen readily activated by the human preparation. None of the aliphatic nitrosamines was activated by the rat or pig. No correlation was observed between efficiency of activation and any of the enzyme activities studied.


Functional domains of a T-DNA promoter active in crown gall tumors.
Promoter domains required for transcriptional expression of the 780 gene of T-right (pTi15955) were identified by deletion mutagenesis. Accurate quantitation of transcriptional activity of a series of 5' and internal deletion mutants was achieved by using a double gene vector containing a reference 780 gene as an internal standard. Results of the 5' deletions delineated an activator element located between -440 and -229 base pairs (bp) from the start of transcription. Removal of this region resulted in a 100-fold decrease in promoter activity. Two relatively small internal deletion/substitution mutations at positions -74 to -76 and -98 to -112 reduced promoter activity to 38 and 42%, respectively. In most cases large-scale internal deletions (38 to 151 bp) occurring in various locations from positions -12 to -348 bp caused a significant loss in major promoter activity. However, three internal deletions starting at position -37 and extending upstream as far as -153 bp either had little effect on transcriptional activity or resulted in increased activity. Removal of the TATA motif drastically reduced promoter activity to less than 0.1% of the wild type. A minor start of transcription was detected 60 bases upstream from the major transcriptional start site. This minor promoter shares the same activator element as the major promoter for full activity. Deletion and insertion mutations downstream of the minor promoter TATA demonstrated the role of the TATA box in positioning the start of transcription.


Emergency Ambulance Utilization in Harlem, New York (July 1985).
Emergency ambulance service in Harlem, New York was studied to determine the degree to which misuse exists. Data were collected prospectively from a paramedic (ALS) ambulance and a basic life support (BLS) ambulance dispatched from Harlem Hospital. The results show that only 29.5% of the 136 calls dispatched to the ALS unit had congruent dispatch and actual priorities (r = 0.05), and only 59.6% of the 255 calls handled by both units were recommended for the unit with the proper level of the skill. 69.9% of calls assigned to the ALS unit were found not to need emergency transport (ET), the most common reason being the "unfounded" call, which represented 35.3% of the sample. ALS intervention was needed on only 20.6% of calls dispatched to the ALS unit. Paramedics were able to correctly access the need for ET 95.6% of the time, but were only willing to refuse transport 35.2% of times in which they felt it was unnecessary. Within the population studied, there exists a significant subset of nonemergency calls which could be more efficiently triaged, both at the level of dispatch and by EMS personnel on the scene. Author Perspective: This research was performed from July 1 to July 26, 1985 to fulfill the requirements of a Community Medicine rotation when the investigator was a student at the Mount Sinai School of Medicine in New York City (presently the Ican School of Medicine at Mount Sinai). Dr. Alexander E. Kuehl was the Vice President of New York City Health and Hospitals Corporation, Emergency Medical Service at the time (prior to New York City EMS moving under the Fire Department) and served as preceptor for the project. This work has not been previously published; however, the investigator was an invited presenter at the "Third International Urban Emergency Medical Service System Symposium" in September 1985 at the World Trade Center in New York City. The oral presentations at the symposium were entitled: "Paramedic Attitudes Concerning On-The-Scene Triage" and "Emergency Ambulance Utilization in Harlem." To complete the work for this 30-day rotation, the investigator performed daily ambulance ride-alongs in Harlem, New York for 15 consecutive days and collected data prospectively from paramedics. EMTs staffing an ambulance during the same time frame provided concurrent information by survey. Some EMS terminology is now outdated; however, the terms have not been updated in order to preserve the historical perspective of the article. While the data are now historical, having been collected nearly 40 years ago, this research provides useful insights into current EMS initiatives, notably: (1) accuracy of assigned dispatch priority, (2) need for emergency transport to an emergency department, and (3) willingness of paramedics to refuse transport in cases where it is deemed unnecessary. The research questions represent many of the challenges that persist in today's EMS world with some being particularly relevant with the advent of Community Paramedicine. Emergency Ambulance Utilization in Harlem truly illustrates "what's old is new again!"


Perioperative Management in Reverse Total Shoulder Arthroplasty.
A successful reverse total shoulder arthroplasty requires careful preoperative planning and perioperative management. Preoperative comorbidity risks, perioperative pain management, and postoperative rehabilitation are all critical components of this arthroplasty. The current review examines available literature to guide the perioperative care of the reverse total shoulder arthroplasty patient. One of the most important advances for shoulder arthroplasty in recent years has been heightened awareness of various modalities for perioperative pain management. A number of recent studies have focused on the options for regional blockade as a critical tool for postoperative pain relief and the use of either continuous interscalene blocks or single shot blocks are supported. Additional studies are necessary to define the best local anesthetic agent and delivery mechanism to provide appropriate pain relief with a low side effect profile. Management of the patient throughout the perioperative course is a critical component in achieving better patient outcomes delivering high quality patient care. An orthopedic surgery team focused on perioperative management is better positioned to decrease adverse events and improve patient outcomes after reverse total shoulder arthroplasty.


Growth and treatment changes in patients treated with a headgear-activator appliance.
The aim of this study was to investigate growth and treatment changes in patients with Class II Division 1 malocclusion treated with a combined headgear-activator appliance. The sample consisted of 20 consecutive male patients treated with headgear-activator. Their compliance was checked after 2 months of treatment, and 3 patients dropped out. Lateral cephalograms, obtained 6 months before treatment, at the start of treatment, after 6 and 12 months of treatment, and 24 months after treatment, were analyzed by the method of Pancherz. The results disclosed that during growth, on average, overjet, molar relationship, and jaw-base relationship improved, and the mandibular incisors became retruded; the changes were statistically significant over a 2-year period. There was no significant change in the overbite. During the 12 months of treatment, there was no maxillary forward growth, and the jaw relationship improved (P <.001) because of forward growth of the mandible (P <.001). The molar relationship and the overbite improved (P <.001). Significant treatment effects included reduction in overjet (P <.001) and overbite (P <.01), and improvement of jaw-base (P <.05) and molar (P <.001) relationships. There was significant restraint of maxillary forward growth (P <.05). The eruption of the maxillary incisors and molars was restrained (P <.05). The average basic growth pattern in Class II Division 1 malocclusion resulted in small favorable changes in overjet, molar relationship, and jaw-base relationship. The main effect of the headgear-activator appliance was a favorable dental change, limited to restraint of maxillary forward growth but not affecting the vertical dimension.


Profiling the enzymatic properties and inhibition of human complement factor B.
Human complement factor B is the crucial catalytic component of the C3 convertase enzyme that activates the alternative pathway of complement-mediated immunity. Although a serine protease in its own right, factor B circulates in human serum as an inactive zymogen and there is a crystal structure only for the inactive state of factor B and various fragments. To provide greater insight to the catalytic function and properties of factor B, we have used short para-nitroanilide derivatives of 4- to 15-residue peptides as substrates to profile the catalytic properties of factor B. Among factors found to influence catalytic activity of factor B was an unusual dependence on pH. Non-physiological alkaline conditions strongly promoted substrate cleavage by factor B, consistent with a pH-accessible conformation of the enzyme that may be critical for catalytic function. Small N-terminal extensions to conventional hexapeptide para-nitroanilide substrates significantly increased catalytic activity of factor B, which was more selective for its cleavage site than trypsin. The new chromogenic assay enabled optimization of catalysis conditions, the profiling of different substrate sequences, and the development of the first reversible and competitive substrate-based inhibitor of factor B. The inhibitor was also shown to prevent in vitro formation of C3a from C3 by factor B, by synthetic and by natural C3 convertase of the alternative complement activation pathway, and to block formation of membrane attack complex. The availability of a reversible substrate-based inhibitor that could stabilize the active conformation of factor B, in conjunction with a pH-promoted higher processing activity, may offer a new avenue to obtain crystal structures of factor B and C3 convertase in an active conformation.


Social Exclusion Index-for Health Surveys (SEI-HS): a prospective nationwide study to extend and validate a multidimensional social exclusion questionnaire.
Social exclusion (SE) refers to the inability of certain groups or individuals to fully participate in society. SE is associated with socioeconomic inequalities in health, and its measurement in routine public health monitoring is considered key to designing effective health policies. In an earlier retrospective analysis we demonstrated that in all four major Dutch cities, SE could largely be measured with existing local public health monitoring data. The current prospective study is aimed at constructing and validating an extended national measure for SE that optimally employs available items. In 2012, a stratified general population sample of 258,928 Dutch adults completed a version of the Netherlands Public Health Monitor (PHM) questionnaire in which 9 items were added covering aspects of SE that were found to be missing in our previous research. Items were derived from the SCP social exclusion index, a well-constructed 15-item instrument developed by the Netherlands Institute for Social Research (SCP). The dataset was randomly divided into a development sample (N =129,464) and a validation sample (N = 129,464). Canonical correlation analysis was conducted in the development sample. The psychometric properties were studied and compared with those of the original SCP index. All analyses were then replicated in the validation sample. The analysis yielded a four dimensional index, the Social Exclusion Index for Health Surveys (SEI-HS), containing 8 SCP items and 9 PHM items. The four dimensions: "lack of social participation", "material deprivation", "lack of normative integration" and "inadequate access to basic social rights", were each measured with 3 to 6 items. The SEI-HS showed adequate internal consistency for both the general index and for two of four dimension scales. The internal structure and construct validity of the SEI-HS were satisfactory and similar to the original SCP index. Replication of the SEI-HS in the validation sample confirmed its generalisability. This study demonstrates that the SEI-HS offers epidemiologists and public health researchers a uniform, reliable, valid and efficient means of assessing social exclusion and its underlying dimensions. The study also provides valuable insights in how to develop embedded measures for public health surveillance.


Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases.
It has been reported that mitochondria can contain up to 12 enzymatic sources of reactive oxygen species (ROS). A majority of these sites include flavin-dependent respiratory complexes and dehydrogenases that produce a mixture of superoxide (O2●-) and hydrogen peroxide (H2O2). Accurate quantification of the ROS-producing potential of individual sites in isolated mitochondria can be challenging due to the presence of antioxidant defense systems and side reactions that also form O2●-/H2O2. Use of nonspecific inhibitors that can disrupt mitochondrial bioenergetics can also compromise measurements by altering ROS release from other sites of production. Here, we present an easy method for the simultaneous measurement of H2O2 release and nicotinamide adenine dinucleotide (NADH) production by purified flavin-linked dehydrogenases. For our purposes here, we have used purified pyruvate dehydrogenase complex (PDHC) and α-ketoglutarate dehydrogenase complex (KGDHC) of porcine heart origin as examples. This method allows for an accurate measure of native H2O2 release rates by individual sites of production by eliminating other potential sources of ROS and antioxidant systems. In addition, this method allows for a direct comparison of the relationship between H2O2 release and enzyme activity and the screening of the effectiveness and selectivity of inhibitors for ROS production. Overall, this approach can allow for the in-depth assessment of native rates of ROS release for individual enzymes prior to conducting more sophisticated experiments with isolated mitochondria or permeabilized muscle fiber.


Intraoperative frozen-section analysis for thyroid nodules: a step toward clarity or confusion?
To determine accuracy and intertest agreement of preoperative fine-needle aspiration cytology (FNAC) and intraoperative frozen-section analysis (FS) findings in thyroid surgery, and to assess the influence of intraoperative FS findings on decision making and the utility of FS in thyroid surgery. Retrospective analysis. The results of preoperative FNAC, intraoperative FS, and final histopathological analyses were taken from the histopathology reports. We calculated intertest agreement using the kappa statistic. Two-hundred fifteen patients who underwent primary thyroid surgery. All patients were treated by the same surgeon (S.J.W.). T he sensitivity and specificity of FNAC were 57.4% and 91.7%, respectively. The sensitivity and specificity of FS were 32.4% and 96.5%, respectively. The intertest agreement was poor (kappa = 0.17). In case of malignant FNAC findings, the FS result did not influence treatment decisions; in case of a malignant FS result on the background of a benign, indeterminate, or nondiagnostic FNAC finding, the FS result influenced treatment decisions in 88% of cases. Intraoperative FS did not give additional information in cases where a malignant neoplasm was predicted by the FNAC finding. In this setting, it led to conflicting results and did not contribute to correct decision making.


The fatty acid composition of phospholipids of spermatozoa from infertile patients.
The lipid composition of the sperm membrane has a significant effect upon the functional characteristics of spermatozoa. In the present study we investigated the fatty acid (FA) composition of subpopulations of spermatozoa separated on a discontinuous Percoll gradient (47:90%) and the FA composition of phospholipids (PL) of sperm heads and tails in both normal and abnormal semen samples. In normozoospermic samples, polyunsaturated fatty acids (PUFA) represented 34.0 +/- 1.3 (mean +/- SE, mole %) and 25.6 +/- 1.2% of total FA of PL of the 47 and 90% Percoll fractions respectively. Docosahexaenoic acid (22:6omega3, DHA) contributed to more than 60% of total PUFA. DHA was significantly lower in both the 47% (P < 0.05) and the 90% (P < 0.01) Percoll fractions of oligozoospermic samples and in the 90% Percoll layer of asthenozoospermic samples (P < 0.01), compared with normozoospermic samples. The omega6/omega3 ratio was significantly increased in both Percoll fractions of samples with oligozoospermia (47%, P < 0.001 and 90%, P < 0.001) or with asthenozoospermia (47%, P < 0.05 and 90%, P < 0.001) compared with normozoospermic samples. The oxidative potential index (OPI) of spermatozoa recovered from the 47% Percoll layer was significantly higher (P < 0.0001) than of those recovered from the 90% Percoll. Mean melting point (MMP), an index of membrane fluidity, was significantly lower in head than in tails (P < 0.01) of spermatozoa, and also in both the 47% (P < 0.01) and 90% (P < 0.001) Percoll fractions of normozoospermic samples in comparison with oligozoospermic samples. The MMP was significantly higher (P < 0.05) in samples of patients with idiopathic oligo/asthenozoospermia, varicocele, and male accessory gland infection (MAGI). These differences in FA composition of PL in subpopulations of human spermatozoa, and in their heads and tails may be related to sperm maturity and to differences in physiological function.


Sulindac metabolites induce caspase- and proteasome-dependent degradation of beta-catenin protein in human colon cancer cells.
Colorectal cancer (CRC) is the second leading cause of cancer death in the USA. Accumulation of beta-catenin protein is nearly ubiquitous in colon adenomas and cancers, presumably due to mutations in the APC or beta-catenin genes that inhibit proteasome-dependent degradation of beta-catenin protein. Substantial clinical, epidemiological, and animal evidence indicate that sulindac and other non-steroidal anti-inflammatory drugs (NSAIDs) prevent the development of CRC. The mechanisms by which sulindac exerts its potent growth inhibitory effects against colon tumor cells are incompletely understood, but down-regulation of beta-catenin has been suggested as one potential mechanism. The goal of this study was to determine the mechanism of beta-catenin protein down-regulation by sulindac metabolites. Treatment of human colon cancer cell lines with apoptotic concentrations of sulindac metabolites (sulindac sulfide, sulindac sulfone) induced a dose- and time-dependent inhibition of beta-catenin protein expression. Inhibition of proteasome activity with MG-132 partially blocked the ability of sulindac sulfide and sulindac sulfone to inhibit beta-catenin protein expression. Pretreatment with the caspase inhibitor z-VAD-fmk blocked morphological signs of apoptosis as well as caspase cleavage, and also partially prevented beta-catenin degradation by sulindac metabolites. These effects occurred in cells with bi-allelic APC mutation (SW480), with wild-type APC but mono-allelic beta-catenin mutation (HCT116) and in cells that lack expression of either COX-1 or -2 (HCT15). These results indicate that loss of beta-catenin protein induced by sulindac metabolites is COX independent and at least partially due to reactivation of beta-catenin proteasome degradation and partially a result of caspase activation during the process of apoptosis.


[Extremes of maternal age and child mortality: analysis between 2000 and 2009].
To analyze the characteristics of infant mortality at the extremes of maternal age. Retrospective, cross-sectional quantitative study using data from Live Birth Certificates, Death Certificates and from Child Death Investigation records in Londrina, Paraná, in the years of 2000-2009. During the 10-year study period, there were 176 infant deaths among mothers up to 19 years of age, and 113 deaths among mothers aged 35 years or more. The infant mortality rate among young mothers was 14.4 deaths per thousand births, compared to 12.9 deaths in the other age group. For adolescent mothers, the following conditions prevailed: lack of a stable partner (p<0.001), lack of a paid job (p<0.001), late start of prenatal care in the second trimester of pregnancy (p<0.001), fewer prenatal visits (p<0.001) and urinary tract infections (p<0.001). On the other hand, women aged 35 or more had a higher occurrence of hypertension during pregnancy (p<0.001), and of surgical delivery (p<0.001). Regarding the underlying cause of infant death, congenital anomalies prevailed in the group of older mothers (p=0.002), and external causes were predominant in the group of young mothers (p=0.019). Both age groups deserve the attention of social services for maternal and child health, especially adolescent mothers, who presented a higher combination of factors deemed hazardous to the child's health.


LoTToR: An Algorithm for Missing-Wedge Correction of the Low-Tilt Tomographic 3D Reconstruction of a Single-Molecule Structure.
A single-molecule three-dimensional (3D) structure is essential for understanding the thermal vibrations and dynamics as well as the conformational changes during the chemical reaction of macromolecules. Individual-particle electron tomography (IPET) is an approach for obtaining a snap-shot 3D structure of an individual macromolecule particle by aligning the tilt series of electron tomographic (ET) images of a targeted particle through a focused iterative 3D reconstruction method. The method can reduce the influence on the 3D reconstruction from large-scale image distortion and deformation. Due to the mechanical tilt limitation, 3D reconstruction often contains missing-wedge artifacts, presented as elongation and an anisotropic resolution. Here, we report a post-processing method to correct the missing-wedge artifact. This low-tilt tomographic reconstruction (LoTToR) method contains a model-free iteration process under a set of constraints in real and reciprocal spaces. A proof of concept is conducted by using the LoTToR on a phantom, i.e., a simulated 3D reconstruction from a low-tilt series of images, including that within a tilt range of ±15°. The method is validated by using both negative-staining (NS) and cryo-electron tomography (cryo-ET) experimental data. A significantly reduced missing-wedge artifact verifies the capability of LoTToR, suggesting a new tool to support the future study of macromolecular dynamics, fluctuation and chemical activity from the viewpoint of single-molecule 3D structure determination.


Physical activity and disordered eating behaviours: Are Caribbean adolescents at risk?
Excessive physical activity (PA) has been linked to increased risk for disordered eating behaviours and eating disorders. This study investigates the relationship between PA and disordered eating behaviours and attitudes (DEBAs) among Jamaican adolescents. This cross-sectional study included 521 adolescents, 12-19 years. Anthropometric measurements were collected, and adolescents completed questionnaires on disordered eating behaviours (EAT-26), physical activity, self-esteem and affect. Associations were assessed using sex-specific mixed-effect linear and logistic regression models. Participants reported exercising an average of 3 days per week. Adolescents who exercised for a longer duration had greater odds of having elevated EAT-26 scores (at least 1 hour-OR = 2.04; 95% CI = 1.03, 4.06; p = .042), while a higher exercise frequency among males (3-5 days per week) was protective against DEBAs (OR 0.38; 95% CI = 0.16, 0.88; p = .025). Female adolescents reported higher prevalence of elevated EAT-26 scores than males (p < .01). Increased negative affect increased odds of an elevated EAT-26 score. Exercise duration and frequency play a role in disordered eating behaviours in Jamaican adolescents and vary by gender. Our findings have implications for weight management interventions and policies, encouraging healthcare providers to monitor PA levels as well as negative affect in adolescents who display disordered eating behaviours.


Liver microsomal inactivation of 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone as an inhibitor of ribonucleotide reductase.
Studies were carried out to determine the effects of preincubation of 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone (MAIQ) with hepatic microsomes on the ability of MAIQ to inhibit CDP reductase activity in vitro. An aliquot from the 100,000 x g supernatant fraction from this incubation was used in the CDP reductase assay. MAIQ incubated in the absence of microsomes inhibited CDP reductase activity in a dose-dependent manner. At high MAIQ concentration (5 microM) CDP reductase activity was inhibited 95%. When MAIQ (5 microM) was first incubated in the presence of hepatic microsomes and NADPH, CDP reductase activity was inhibited only 30%. This attenuation of MAIQ inhibition was dependent on time of incubation and microsomal protein concentration and showed an obligatory requirement for NADPH or NADH. Significant attenuation was observed at pyridine nucleotide concentrations as low as 0.1 mM. Heat denaturation of microsomal proteins inactivated their ability to attenuate the MAIQ inhibition. Microsomes prepared from Ehrlich tumor cells were ineffective as inactivators of MAIQ. Results of our studies show that hepatic microsomes contain an enzyme(s) which can inactive MAIQ as an inhibitor of CDP reductase.


Expression of the zebrafish Iroquois genes during early nervous system formation and patterning.
Iroquois genes are involved in many patterning processes during development. In particular, they act as prepattern genes to control proneural gene expression both in Drosophila and in vertebrates. In this paper, we have analyzed the expression during embryogenesis of the 11 zebrafish Iroquois genes, with special interest for nervous system formation and patterning. During the first 2 days of development, Iroquois genes are expressed in distinct domains in the neuroepithelium, as well as in groups of neuronal progenitors and neurons. They are also expressed at different stages of placodal development. These expression patterns are similar to the patterns of the murine irx genes and also show features specific to teleosts. For the zebrafish Iroquois gene family, we find both specific patterns and patterns conserved within a cluster, between paralogues, or in most genes of the family. Overall, these expression data suggest functions for the Iroquois family of transcription factors in neural and placodal patterning, neurogenesis, and neuronal specification.


A CRISPR/Cas12a-responsive dual-aptamer DNA network for specific capture and controllable release of circulating tumor cells.
The separation and detection of circulating tumor cells (CTCs) have a significant impact on clinical diagnosis and treatment by providing a predictive diagnosis of primary tumors and tumor metastasis. But the responsive release and downstream analysis of live CTCs will provide more valuable information about molecular markers and functional properties. To this end, specific capture and controllable release methods, which can achieve the highly efficient enrichment of CTCs with strong viability, are urgently needed. DNA networks create a flexible, semi-wet three-dimensional (3D) microenvironment for cell culture, and have the potential to minimize the loss of cell viability and molecular integrity. More importantly, responsive DNA networks can be reasonably designed as smart sensors and devices to change shape, color, disassemble, and giving back to external stimuli. Here, a strategy for specifically collecting cells using a dual-aptamer DNA network is designed. The proposed strategy enables effective capture, 3D encapsulation, and responsive release of CTCs with strong viability, which can be used for downstream analysis of live cells. The programmability of CRISPR/Cas12a, a powerful toolbox for genome editing, is used to activate the responsive release of captured CTCs from the DNA network. After activation by a specified double-strand DNA (dsDNA) input, CRISPR/Cas12a cleaves the single-stranded DNA regions in the network, resulting in molecular to macroscopic changes in the network. Accompanied by the deconstruction of the DNA network into fragments, controllable cell release is achieved. The viability of released CTCs is well maintained and downstream cell analysis can be performed. This strategy uses the enzymatic properties of CRISPR/Cas12a to design a platform to improve the programmability and versatility of the DNA network, providing a powerful and effective method for capturing and releasing CTCs from complex physiological samples.


Evaluation of a Pilot Project to Introduce Simulation-Based Team Training to Pediatric Surgery Trauma Room Care.
Introduction. Several studies in pediatric trauma care have demonstrated substantial deficits in both prehospital and emergency department management. Methods. In February 2015 the PAEDSIM collaborative conducted a one and a half day interdisciplinary, simulation based team-training course in a simulated pediatric emergency department. 14 physicians from the medical fields of pediatric surgery, pediatric intensive care and emergency medicine, and anesthesia participated, as well as four pediatric nurses. After a theoretical introduction and familiarization with the simulator, course attendees alternately participated in six simulation scenarios and debriefings. Each scenario incorporated elements of pediatric trauma management as well as Crew Resource Management (CRM) educational objectives. Participants completed anonymous pre- and postcourse questionnaires and rated the course itself as well as their own medical qualification and knowledge of CRM. Results. Participants found the course very realistic and selected scenarios highly relevant to their daily work. They reported a feeling of improved medical and nontechnical skills as well as no uncomfortable feeling during scenarios or debriefings. Conclusion. To our knowledge this pilot-project represents the first successful implementation of a simulation-based team-training course focused on pediatric trauma care in German-speaking countries with good acceptance.


Ghrelin, acylated ghrelin, leptin and PYY-3 levels in hyperemesis gravidarum.
To assess the serum levels of gut and adipocyte-derived metabolic hormones that control appetite, adipocity, weight gain and energy hemostasis, namely total ghrelin (TG), acylated ghrelin (AG), leptin and PYY-3 in hyperemesis gravidarum (HG). Plasma samples of 86 women in their first trimester pregnancies with HG (n = 30), morning sickness of pregnancy (MSP) (n = 34) and control (n = 22) groups were obtained. Serum levels of TG, AG, leptin and PYY-3 were compared between the groups, and the correlations with severity of symptoms using modified PUQE (Pregnancy Unique Quantification of Emesis) scoring, BMI, E2, hCG and TSH were calculated. Levels of TG, AG, PYY-3 and the TG/leptin ratio were significantly higher in HG group compared to MSP and the control groups (p ≤ 0.017). AG/TG ratio was significantly lower in the HG group compared to both MSP and control groups (p ≤ 0.017). There were either weak or no significant statistical correlations between the gut and adipose-related hormones and the modified PUQE scores, BMI, E2, hCG and TSH. Gut-derived metabolic hormones ghrelin, AG and PYY-3 may be involved in the HG pathophysiology.


Associations of genetic variation in CASP3 gene with noise-induced hearing loss in a Chinese population: a case-control study.
Noise-induced hearing loss (NIHL) is a complex disease caused by environmental and genetic risk factors. This study explored the relationship between the genetic variations in the CASP gene and the risk of developing NIHL among Chinese workers exposed to occupational noise. A case-control study of 272 NIHL workers and 272 normal-hearing workers matched for age, sex and years of noise exposure was conducted. Fifteen single-nucleotide polymorphisms (SNP) in the CASP1, CASP3, CASP4, CASP5, CASP6, CASP8, CASP9, CASP10 and CASP14 genes were genotyped using the polymerase chain reaction-ligase detection reaction method. Using conditional logistic regression models, the adjusted odds ratios (ORs) and 95% confidence intervals (CIs) of genetic variations associated with NIHL risk were calculated. Two SNPs in the CASP3 gene were associated with NIHL risk. For rs1049216, TT genotype was associated with a decreased risk of NIHL (OR = 0.246, 95% CI = 0.069-0.886) when compared with the CC genotype. For rs6948, the AC and CC genotype were associated with a decreased NIHL risk (OR = 0.568, 95% CI = 0.352-0.916) compared with AA genotype. There were joint effects of working time and CASP3 polymorphisms on NIHL risk (P < 0.05). Genetic variations in the CASP3 gene and the joint effects of working time and CASP3 polymorphisms may modify the risk of developing NIHL.


Phytotherapeutic potential of natural herbal medicines for the treatment of mild-to-severe atopic dermatitis: A review of human clinical studies.
For many decades, natural herbal medicines, polyherbal formulations and/or decoctions of plant-derived materials have widely been accepted as alternative complementary therapies for the treatment, cure or prevention of a wide range of acute and chronic skin diseases including chronic herpes, prurigo, acute and chronic wounds, psoriasis and atopic dermatitis (AD). This review was aimed to summarize and critically discuss about the therapeutic viability and clinical applicability of natural herbal medicines for the treatment of AD in human. The critical analysis of the literature revealed that oral (in the form of capsules, syrup or granules) and/or topical application (alone or in conjunction with wet-wrap dressing and/or acupuncture) of natural herbal medicines exhibit remarkable potential for the treatment of mild-to-severe AD in adults, children, infants and in the pregnant women. In this review, the clinical efficacy of various herbal formulations such as Chinese herbal therapies, Korean medicines, Iranian medicines, honey, natural herbal oils (coconut oil, olive oil and mineral oil), beeswax, dodder seeds and whey for the treatment of AD has been discussed. The clinical anti-AD efficacy of these complementary therapies has been observed in terms of down-regulation in Scoring Atopic Dermatitis (SCORAD) index, erythematic intensity, Children's Dermatology Life Quality Index (CDLQI), Dermatology Life Quality Index (DLQI), pruritus and itching frequency, transepidermal water loss (TEWL) and expression of AD-mediated chemokines. Conclusively, we recognized that natural herbal medicines demonstrate remarkable clinical efficacy when used alone or in conjunction with other complementary therapies for the treatment of AD in patients of all ages as well as pregnant women.


Association of Programmed Death 1 Protein Ligand (PD-L1) Expression With Prognosis in Merkel Cell Carcinoma.
Background: Merkel cell carcinoma (MCC) is a rare neuroendocrine skin cancer. Prior to the advent of immunotherapy, treatment options were limited. In our study, we evaluate the impact of tumor cell PD-L1 expression and tumor immune microenvironment on survival in MCC patients who were not treated with immune checkpoint inhibitors. Methods: Clinical data and tissue samples were collected from 78 patients with confirmed MCC treated at Dana-Farber Cancer Institute. Specimens were analyzed for the distribution of PD-L1 by immunohistochemistry staining (IHC) and standardized analysis. Results were correlated with survival data. Results: In this study, membrane and cytoplasmic MCC tumor cell staining for PD-L1 was detected in 22.4% (15 of 67) of cases and PD-L1 staining of intratumoral microvessels and PD-L1 positive immune cells at the infiltrative margins of the tumor in 92.5% (62 of 67) of cases. In patients untreated with immune checkpoint inhibitors, median overall survival was not different for patients based on PD-L1 expression (PD-L1+ 64 months vs. PD-L1- not reached; HR = 1.26, 95% CI: 0.46-3.45; p = 0.60). Conclusion: PD-L1 expression is frequently detected in MCC tumor cells and tumor microenvironment. PD-L1 expression did not affect prognosis in this cohort that had not received PD-1/L1 blockade.


The Relationship between Human Embryo Parameters and De Novo Chromosomal Abnormalities in Preimplantation Genetic Testing Cycles.
In total, 456 PGT cycles, including 283 PGT-SR cycles and 173 PGT-A cycles, were assessed through comprehensive chromosome screening (CCS) from January 2017 to June 2020 at the Department of Reproductive Medicine of the Third Affiliated Hospital of Zhengzhou University. Trophectoderm (TE) biopsies were sequenced using next-generation sequencing (NGS). The incidence of de novo chromosome abnormalities was calculated, and the relationships between de novo chromosome abnormality rates and maternal age, number of oocytes retrieved, and parameters of cleavage-stage embryos and blastocyst-stage embryos were investigated. The incidence of de novo chromosome abnormalities was 28.0% (318/1,135) in the PGT-SR cycles and 36.3% (214/590) in the PGT-A cycles, which increased with maternal age in both PGT-SR cycles (P = 0.018) and PGT-A cycles (P < 0.001). The incidence of de novo chromosome abnormalities was related to TE grade (P < 0.001), internal cell mass grade (P = 0.002), and development speed (day 5 vs. day 7: P < 0.001) of blastocyst-stage embryos. The incidence of de novo chromosomal abnormalities was irrelevant to the number of oocytes retrieved and the parameters of the embryo at the cleavage stage. Blastocysts with higher morphology scores and faster progression had a lower incidence of de novo chromosome abnormalities, especially complex chromosome abnormalities. De novo chromosome abnormalities may negatively affect the morphological grading of blastocysts. Our findings will provide valuable information to the fertility doctor for embryo selection in non-PGT cycles.


The effects on anxiety and quality of life of breast cancer patients following completion of the first cycle of chemotherapy.
Breast cancer patients as part of their treatment need to undergo various forms of chemotherapy. This is considered as a burdensome experience for many patients often leading to significant levels of anxiety. The aim of the study was to explore the anxiety levels and any correlations to the quality of life of women with breast cancer that were undergoing chemotherapy. This was a cross-sectional study utilizing an explanatory sequential design. Data were collected from 355 women with breast cancer with the Self Anxiety Scale, the EORTC QLQ-C30, the EORTC QLQ-BR23 and sociodemographic questionnaires. Further insight to patients' experiences was given through 12 in-depth interviews. Anxiety scores ranged between 24 and 75 (45.7 ± 10.11), with 44% reporting serious or/and intense anxiety. The results revealed statistically significant differences on patients' anxiety levels depending on their source of support. Overall, patients' global health-related quality of life was found to be low to average 55.91 ± 17.94. The results showed low emotional functioning (49.30 ± 29.12), low role functions (56.34 ± 27.50) and low sexual functioning (24.93 ± 20.75). Patients also reported experiencing problems with fatigue (49.04 ± 29.12), insomnia (44.32 ± 32.97), hair loss (48.25 ± 38.32) and arm symptoms (36.53 ± 23.71). Patients being solely supported by the family experienced higher anxiety levels (p < 0.001) and lower quality of life (p < 0.001). There was a statistically significant negative correlation between anxiety and quality of life (r = -0.623, p < 0.001). Statistically significant differences were also found in relation to demographics, anxiety and quality of life. The interviews provided further evidence on the impact of anxiety on patients' lives. The time following the completion of the first cycle of chemotherapy is associated with anxiety and lower quality of life levels in breast cancer patients. Healthcare providers should consider the supportive healthcare needs from the beginning of chemotherapy in patients to optimize their conventional and supportive healthcare outcomes.


Comparative in vitro degradation of the human hemorphin LVV-H7 in mammalian plasma analysed by capillary zone electrophoresis and mass spectrometry.
The human hemorphin LVV-H7 (L32VVYPWTQRF41) is a hemoglobin-beta, -gamma, -delta or -epsilon chain derived cationic decapeptide of the micro-opioid receptor binding family. It exhibits potential pharmacological value relevant, for example, for blood pressure regulation, learning performance and Alzheimer's disease. The regulatory potency is strictly dependent on the length of the amino acid sequence which is sensitive towards proteinases from tissues and plasma. To analyse LVV-H7 in vitro degradation in mammalian plasma, a novel multi-component quantitative capillary zone electrophoretic (CZE) procedure was applied, combined with qualitative metabolite profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In all types of plasma, LVV-H7 was N-terminally truncated generating four metabolites (M1-M4) with an intact C-terminus: M1 (V33VYPWTQRF41), M2 (V34YPWTQRF41), M3 (Y35PWTQRF41) and M4 (W37TQRF41). In EDTA plasma these degradation products were detected exclusively, whereas in citrate and heparin plasma four further metabolites appeared resulting from additional C-terminal cleavage of the dipeptide R40F41: M5 (L32VVYPWTQ39), M6 (V33VYPWTQ39), M7 (V34YPWTQ39) and M8 (Y35PWTQ39). In the presence of selective proteinase inhibitors aminopeptidase M and angiotensin-converting enzyme (for N- and C-terminal truncation, respectively) were identified as plasma enzymes responsible for hemorphin degradation. Furthermore, striking inter-mammalian species distinctions were detected revealing strongly differing degradation velocities but similar metabolite patterns.


Photoreactions of 3-diazo-3H-benzofuran-2-one; dimerization and hydrolysis of its primary photoproduct, a quinonoid cumulenone: a study by time-resolved optical and infrared spectroscopy.
Light-induced deazotization of 3-diazo-3H-benzofuran-2-one (1) in solution is accompanied by facile (CO)-O bond cleavage yielding 6-(oxoethenylidene)-2,4-cyclohexadien-1-one (3), which appears with a rise time of 28 ps. The expected Wolff-rearrangement product, 7-oxabicyclo[4.2.0]octa-1,3,5-trien-8-ylidenemethanone (4), is not formed. The efficient light-induced formation of the quinonoid cumulenone 3 opens the way to determine the reactivity of a cumulenone in solution. The reaction kinetics of 3 were monitored by nanosecond flash photolysis with optical (lambda(max) approximately 460 nm) as well as Raman (1526 cm(-1)) and IR detection (2050 cm(-)(1)). Remarkably, the reactivity of 3 is that expected from its valence isomer, the cyclic carbene 3H-benzofuran-2-one-3-ylidene, 2. In aqueous solution, acid-catalyzed addition of water forms the lactone 3-hydroxy-3H-benzofuran-2-one (5). The reaction is initiated by protonation of the cumulenone on its beta-carbon atom. In hexane, cumulenone 3 dimerizes to isoxindigo ((E)-[3,3']bibenzofuranylidene-2,2'-dione, 7), coumestan (6H-benzofuro[3,2-c][1]benzopyran-6-one, 8), and a small amount of dibenzonaphthyrone ([1]benzopyrano[4,3-][1]benzopyran-5,11-dione, 9) at a nearly diffusion-controlled rate. Ab initio calculations (G3) are consistent with the observed data. Carbene 2 is predicted to have a singlet ground state, which undergoes very facile, strongly exothermic (irreversible) ring opening to the cumulenone 3. The calculated barrier to formation of 4 (Wolff-rearrangement) is prohibitive. DFT calculations indicate that protonation of 3 on the beta-carbon is accompanied by cyclization to the protonated carbene 2H(+), and that dimerization of 3 to 7 and 9 takes place in a single step with negligible activation energy.


Conformation of peptides in lipid membranes studied by x-ray grazing incidence scattering.
Although the antimicrobial, fungal peptide alamethicin has been extensively studied, the conformation of the peptide and the interaction with lipid bilayers as well as the mechanism of channel gating are still not completely clear. As opposed to studies of the crystalline state, the polypeptide structures in the environment of fluid bilayers are difficult to probe. We have investigated the conformation of alamethicin in highly aligned stacks of model lipid membranes by synchrotron-based x-ray scattering. The (wide-angle) scattering distribution has been measured by reciprocal space mappings. A pronounced scattering signal is observed in samples of high molar peptide/lipid ratio which is distinctly different from the scattering distribution of an ideal helix in the transmembrane state. Beyond simple models of ideal helices, the data is analyzed in terms of models based on atomic coordinates from the Brookhaven Protein Data Bank, as well as from published molecular dynamics simulations. The results can be explained by assuming a wide distribution of helix tilt angles with respect to the membrane normal and a partial insertion of the N-terminus into the membrane.


The lateral lesser toe fillet flap for diabetic foot soft tissue closure: surgical technique and case report.
Wound closure for the diabetic foot can be challenging and often involves amputation or reconstruction. The authors describe a surgical technique and a case report of lateral lesser toe fillet flap in the management of a diabetic foot wound. The lateral lesser toe fillet flap reconstruction is a reproducible technique that incurs comparatively minimal technical complexity and provides a favorable option in the management of diabetic foot wounds where soft tissue coverage is required.


Approaches to polycyclic 1,4-dioxygenated xanthones. Application to total synthesis of the aglycone of IB-00208.
Hexacyclic xanthone natural products such as IB-00208 present a formidable challenge in organic synthesis. A new approach to polycyclic 1,4-dioxygenated xanthones from benzocyclobutenones has been developed and applied to the first total synthesis of the aglycone of IB-00208. The 22-step synthesis features an acetylide stitching process that joins an aryl aldehyde with an angularly fused benzocyclobutenone, which was prepared by a ring-closing metathesis reaction. The resulting acetylenic benzocyclobutenone diol underwent a Moore rearrangement to give an intermediate that was further elaborated to the aglycone of IB-00208 as a mixture of hydroquinone-quinone tautomers.


Quantitative assessment of microenvironment characteristics and metabolic activity in glioma via multiphoton microscopy.
Tumor microenvironment and metabolic activity in gliomas are the important biomarkers to evaluate the progression of gliomas. Many evidences have suggested that the targeting of metabolic activity and tumor microenvironment simultaneously can be more effective to take the tumor therapy. Therefore, the noninvasive, accurate assessment of tumor microenvironment and metabolic activity is quite important in clinical practice. Multiphoton microscopy (MPM), based on two-photon-excited fluorescence and second harmonic generation was performed on unstained glioma tissues. With our combined image analysis approaches, our research findings indicate that MPM is able to qualitatively and quantitatively describe the microenvironment characteristics in gliomas, such as collage deposition in extracellular matrix, lymphocyte infiltration and tumor angiogenesis, etc. Meanwhile, the metabolic activity can also be quantitatively evaluated by optical redox ratio, NADH and FAD intensity. With the microendoscope and fiberscope are portable, MPM technique can be used to perform in-vivo studies and clinical examinations in gliomas.


Solution-processed squaraine bulk heterojunction photovoltaic cells.
The donor, 2,4-bis[4-(N,N-diisobutylamino)-2,6-dihydroxyphenyl] squaraine (SQ) is used with the acceptor, [6,6]-phenyl C70 butyric acid methyl ester (PC70BM) to result in efficient, solution-processed, small-molecule bulk heterojunction photovoltaic cells. The distribution of the donor nanoparticles in the acceptor matrix as a function of relative concentrations results in a trade-off between exciton dissociation and hole mobility (and hence, cell series resistance). A bulk heterojunction solar cell consisting of an active region with a component ratio of SQ to PC70BM of 1:6 has a power conversion efficiency of 2.7 +/- 0.1% with a 8.85 +/- 0.22 mA/cm(2) short-circuit current density and an open-circuit voltage of 0.89 +/- 0.01 V obtained under simulated 1 sun (100 mW/cm(2)) air mass 1.5 global (AM1.5 G) solar illumination. This is a decrease from 3.3 +/- 0.3% at 0.2 sun intensity, and is less than that of a control planar heterojunction SQ/C60 cell with 4.1 +/- 0.2% at 1 sun, suggesting that the nanoparticle morphology introduces internal resistance into the solution-based thin film. The nanomorphology and hole mobility in the films is strongly dependent on the SQ-to-PC70BM ratio, increasing by greater than 2 orders of magnitude as the ratio increases from 28% to 100% SQ.


Impact of environmental microbiota on human microbiota of workers in academic mouse research facilities: An observational study.
To characterize the microbial environment of workers in academic mouse research facilities using endotoxin, 16S qPCR, and 16S amplicon sequencing. To determine whether the work microbiome contributes to the human microbiome of workers. We performed area air sampling from the animal rooms, dirty, middle, and setup cage wash locations in four academic mouse research facilities. 10 workers in the dirty cage wash area underwent personal air sampling as well as repeated collection of nasal, oral, and skin samples before and after the work shift. Environmental samples underwent measurement of endotoxin, mouse allergen, bacteria copy number via 16S qPCR, and microbial identification via 16S rDNA sequencing. 16S rDNA sequencing was also performed on human samples before and after the work shift. SourceTracker was used to identify the contribution of the work microbiome to the human microbiome. Median endotoxin levels ranged from undetectable to 1.0 EU/m3. Significant differences in mouse allergen levels, bacterial copy number, microbial richness, and microbial community structure were identified between animal, dirty, middle, and setup cage wash locations. Endotoxin levels had only a moderate correlation with microbial composition. Location within a facility was a stronger predictor of microbial community composition (R2 = 0.41, p = 0.002) than facility. The contribution of the work microbiome to the pre-shift human microbiome of workers was estimated to be 0.1 ± 0.1% for the oral microbiome; 3.1 ± 1.9% for the nasal microbiome; and 3.0 ± 1.5% for the skin microbiome. The microbial environment of academic animal care facilities varies significantly by location rather than facility. Endotoxin is not a proxy for assessment of environmental microbial exposures using 16S qPCR or 16S rDNA sequencing. The work microbiome contributes to the composition of the nasal and skin microbiome of workers; the clinical implications of this observation should be further studied.


The Use of Anatomical Dissection Videos in Medical Education.
Dissection videos are commonly utilized in gross anatomy courses; however, the actual usage of such videos, as well as the academic impact of student use of these videos, is largely unknown. Understanding how dissection videos impact learning is important in making curricular decisions. In this study, 22 dissection videos were created to review structures identified in laboratory sessions throughout the Organ Systems 1 (OS1), 2 (OS2), and 3 (OS3) courses. Dissection videos were provided to 201 first-year medical students, and viewing data were recorded. Demographic data for age and gender identity were also collected from students. Overall, there was a significant decrease in total views (P = 0.001), the number of students who pressed play (P < 0.001), and the number of students who viewed ≥ 90% of the total length of videos (P < 0.001) from OS1 to OS3. The total adjusted time spent viewing videos was not significantly different between individual OS courses. There were some instances where significant differences existed in examination performance between those who did and did not view videos, and by time spent viewing videos. There were no significant differences in time spent viewing videos by gender. Together these data suggest that students may utilize dissection videos more at the beginning of a dissection course, although they remain an important resource throughout the year for a subset of students.