ATP-sensitive K+ channels, adenosine, and nitric oxide-mediated mechanisms account for coronary vasodilation during exercise.
We previously reported that combined blockade of adenosine receptors and ATP-sensitive K+ channels (K+(ATP) channels) blunted but did not abolish the response of coronary blood flow to exercise. This study tested the hypothesis that the residual increase in coronary flow in response to exercise after adenosine receptor and K+(ATP) channel blockade is dependent on endogenous NO. Dogs were studied at rest and during a four-stage treadmill exercise protocol under control conditions, during K+(ATP) channel blockade with glibenclamide (50 microg x kg(-1) x min(-1) i.c.) in the presence of adenosine receptor blockade with 8-phenyltheophylline (8-PT, 5 mg/kg i.v.), and after the addition of the NO synthase inhibitor N(G)-nitro-L-arginine (LNNA, 1.5 mg/kg i.c.). During control conditions, coronary blood flow was 49 +/- 3 mL/min at rest and increased to 92 +/- 8 mL/min at peak exercise. LNNA alone or in combination with 8-PT did not alter resting coronary flow and did not impair the normal increase in flow during exercise, indicating that when K+(ATP) channels are intact, neither NO nor adenosine-dependent mechanisms are obligatory for maintaining coronary blood flow. Combined K+(ATP) channel and adenosine blockade decreased resting coronary flow to 27 +/- 3 mL/min (P<.05), but exercise still increased flow to 45 +/- 5 mL/min (P<.05). The subsequent addition of LNNA further decreased resting coronary flow to 20 +/- 2 mL/min and markedly blunted exercise-induced coronary vasodilation (coronary vascular conductance, 0.20 +/- 0.03 mL x min(-1) x mm Hg(-1) at rest versus 0.24 +/- 0.04 mL x min(-1) x mm Hg(-1) during the heaviest level of exercise; P=.22), so that coronary flow both at rest and during exercise was below the control resting level. The findings suggest that K+(ATP) channels are critical for maintaining coronary vasodilation at rest and during exercise but that when K+(ATP) channels are blocked, both adenosine and NO act to increase coronary blood flow during exercise. In the presence of combined K+(ATP) channel blockade and adenosine receptor blockade, NO was able to produce approximately one quarter of the coronary vasodilation that occurred in response to exercise when all vasodilator systems were intact.


A Cancer and Leukemia Group B phase II study of sunitinib malate in patients with previously treated metastatic pancreatic adenocarcinoma (CALGB 80603).
The Cancer and Leukemia Group B (CALGB) conducted a phase II study evaluating sunitinib in patients with progressive metastatic pancreas adenocarcinoma following prior gemcitabine-based therapy (trial CALGB 80603; ClinicalTrials.gov identifier, NCT00397787). The primary endpoint was to determine the disease control rate (DCR) as measured by the Response Evaluation Criteria in Solid Tumors (complete response, partial response [PR], and stable disease) at 6 weeks. Patients aged ≥18 years with an Eastern Cooperative Oncology Group (ECOG) performance status score of 0-2 and with progressive pancreas adenocarcinoma following treatment with gemcitabine were eligible. Sunitinib was dosed at 50 mg orally days 1-28, every 42 days (1 cycle). The statistical plan called for a three-stage design. A DCR ≥15% was considered worthy of further study. In total, 77 patients were enrolled. Forty-two (54.6%) enrollees were male. The median age was 65 years. The ECOG performance status score distribution was: 0, 39%; 1, 50%; 2, 11%. The DCR was 21.6%; one patient (1.4%) had a PR and 15 patients (20.3%) had stable disease as their best response. The progression-free survival time was 1.31 months (95% confidence interval [CI] 1.25-1.38 months) and overall survival time was 3.68 months (95% CI, 3.06-4.24 months). The study met its primary endpoint; however sunitinib had minimal activity and moderate toxicity in a population of gemcitabine-refractory pancreas adenocarcinoma patients. For future studies, limiting enrollment to patients with an ECOG performance status score of 0-1 is recommended.


Pathology of fatal West Nile virus infections in native and exotic birds during the 1999 outbreak in New York City, New York.
West Nile fever caused fatal disease in humans, horses, and birds in the northeastern United States during 1999. We studied birds from two wildlife facilities in New York City, New York, that died or were euthanatized and were suspected to have West Nile virus infections. Using standard histologic and ultrastructural methods, virus isolation, immunohistochemistry, in situ hybridization and reverse-transcriptase polymerase chain reaction, we identified West Nile virus as the cause of clinical disease, severe pathologic changes, and death in 27 birds representing eight orders and 14 species. Virus was detected in 23/26 brains (88%), 24/ 25 hearts (96%), 15/18 spleens (83%), 14/20 livers (70%), 20/20 kidneys (100%), 10/13 adrenals (77%), 13/ 14 intestines (93%), 10/12 pancreata (83%), 5/12 lungs (42%), and 4/8 ovaries (50%) by one or more methods. Cellular targets included neurons and glial cells in the brain, spinal cord, and peripheral ganglia; myocardial fibers; macrophages and blood monocytes; renal tubular epithelium; adrenal cortical cells; pancreatic acinar cells and islet cells; intestinal crypt epithelium; oocytes; and fibroblasts and smooth muscle cells. Purkinje cells were especially targeted, except in crows and magpies. Gross hemorrhage of the brain, splenomegaly, meningoencephalitis, and myocarditis were the most prominent lesions. Immunohistochemistry was an efficient and reliable method for identifying infected cases, but the polyclonal antibody cross-reacted with St. Louis encephalitis virus and other flaviviruses. In contrast, the in situ hybridization probe pWNV-E (WN-USAMRIID99) reacted only with West Nile virus. These methods should aid diagnosticians faced with the emergence of West Nile virus in the United States.


Periodontal health and treatment needs among hospitalized chronic psychiatric patients in Istanbul, Turkey.
The aim of the study was to evaluate the periodontal health and treatment needs of chronically hospitalized psychiatric patients in Istanbul, Turkey. The subjects' periodontal health was recorded by the CPI (Community Periodontal Index) method. Of the 330 patients examined, 179 (52.5%) were males and 151 (47.5%) females. The mean age of the patients was 49.2 +/- 11.7 years. The majority (61.8%) was diagnosed with schizophrenia and 30.6% diagnosed with mental retardation. The mean length of hospitalization was 16.0 +/- 10.9 years. Healthy periodontal tissues (CPI 0) were found in 8.8% of the subjects. Bleeding on probing (CPI 1) was recorded in 6.3%, and dental calculus (CPI 2) in 51.8% of the subjects. These were determined as the worst findings. Altogether, 33% of the subjects had deep periodontal pockets, 14.2% with at least one 4- to 5-mm pocket (CPI 3), and 18.8% with at least one 6-mm pocket (CPI 4). The stepwise logistic regression analysis, between the final CPI score and seven variables including age, gender, psychiatric diagnosis, length of hospitalization, degree of helplessness, tooth brushing habits and smoking, showed that irregular tooth brushing habits and male gender were significant contributors to having a final CPI score of 2 or more. The regression analysis also showed that tooth brushing habits remained as an explanatory variable in CPI 0 coded subjects; helplessness and psychiatric diagnosis (mental retardation) in CPI 2; tooth brushing habits and psychiatric diagnosis (schizophrenia) in CPI 3; and only helplessness in CPI 4. The present study underlines a considerable need for prevention and treatment of periodontal disease among chronic psychiatric patients in Istanbul. Efforts need to be focused above all on raising this population's awareness of the importance of oral hygiene and on early diagnosis of periodontal problems.


Implications of the new definition of myocardial infarction.
The new definition of acute myocardial infarction is based primarily on raised troponin levels because of the sensitivity and specificity of these markers and their correlation with the pathophysiology of acute coronary syndromes with plaque fissuring or rupture and embolisation of platelets causing myocyte necrosis. Raised troponin levels are associated with increased risks of death and recurrent myocardial infarction. Greater treatment benefit with low molecular weight heparin, IIb/IIIa antagonists and revascularisation is seen when troponin levels are raised. There are many implications for patients and society of the new definition including changes in insurability and ability to continue certain occupations. Many more patients, who would previously been diagnosed as having unstable angina, will now be diagnosed as having had an acute myocardial infarction. In addition case fatality rates will fall and comparison with previous epidemiological studies using the old definition will be problematic. However, the new definition may result in greater use of evidence based therapies with improved patient outcomes and decreased community death rates.


Examining dietary self-talk content and context for discretionary snacking behaviour: a qualitative interview study.
Background: Consuming discretionary snack foods high in calories, salt, sugar or fat in between regular meals can have a negative impact on weight management and health. Despite the intention to refrain from discretionary snacking, individuals often report feeling tempted by snack foods. A cognitive process to resolve food choice related tension may be dietary self-talk which is one's inner speech around dietary choice. This study aimed to understand the content and context of dietary self-talk before consuming discretionary snack foods. Methods: Qualitative semi-structured interviews based on Think-Aloud methods were conducted remotely. Participants answered open-ended questions and were presented with a list of 37 dietary self-talk items. Interview transcripts were analyzed thematically. Results: Interviews (n = 18, age: 19-54 years, 9 men, 9 women) confirmed the frequent use of dietary self-talk with all 37 content items endorsed. Reported use was highest for the self-talk items: 'It is a special occasion'; 'I did physical activity/exercise today'; and 'I am hungry'. Three new items were developed, eight items were refined. Identified key contextual themes were: 'reward', 'social', 'convenience', 'automaticity', and 'hunger'. Conclusions: This study lists 40 reasons people use to allow themselves to consume discretionary snack foods and identifies contextual factors of dietary-self talk. All participants reported using dietary self-talk, with variation in content, frequency and degree of automaticity. Recognising and changing dietary self-talk may be a promising intervention target for changing discretionary snacking behaviour.


The association between therapeutic plasma exchange and the risk of mortality among patients critically ill with COVID-19: a meta-analysis.
Background: Cytokine storm has been widely known to contribute to the development of the critical condition in patients with coronavirus disease 2019 (COVID-19), and studies had been conducted to assess the potential aspect of cytokine storm elimination by performing therapeutic plasma exchange (TPE). However, contradictory findings were observed. The objective of this study was to assess the association between TPE and the reduction of mortality of critically ill COVID-19 patients. Methods: A meta-analysis was conducted by collecting data from PubMed, Scopus, and Web of Science. Data on the mortality rate of critically ill COVID-19 patients treated with TPE plus standard of care and that of patients treated with standard of care alone were analyzed using a Z test. Results: We included a total of four papers assessing the association between TPE and the risk of mortality among critically ill COVID-19 patients. Our findings suggested that critically ill COVID-19 patients treated with TPE had lower risk of mortality compared to those without TPE treatment. Conclusion: Our study has identified the potential benefits of TPE in reducing the risk of mortality among critically ill COVID-19 patients.


Ultrastructure of the cell types of the anterior hypophysis in a lizard. I. Corticotrophs.
Corticotrophs of the teiid lizard Cnemidophorus lemniscatus are situated in the rostral zone of the pars distalis. In normal animals, they are usually rounded cells with slightly eccentric vesicular nuclei, especially characterized by a lucent hyaloplasm and medium-sized secretory granules of uniform high density. Granules are almost spherical, with small angular deformations, and closely bounded by a fuzzy membrane. Many cells have only a few or a moderate number of granules, with large areas of cytoplasm devoid of them; in others, granules fill the supranuclear region. The cytoplasm exhibits numerous ribosomes, often in rosettes and mostly free, a series of loosely superimposed cisternae of rough endoplasmic reticulum, small dictyosomes, and elongate mitochondria of light matrix. Metyrapone administration during 2-8 days causes dramatic alterations in corticotrophs; they become hypertrophic and extensively degranulated, with a great development of the endoplasmic reticulum and Golgi apparatus, eventually showing a row of large peripheral granules of uneven structure, enclosed in ample vesicles studded with ribosomes. A lesser degree of hypertrophy and degranulation of corticotrophs appears during the first two weeks after thyroidectomy or gonadectomy, and may be partially attributed to surgical stress. Well granulated enlarged corticotrophs, with hypertrophic endoplasmic reticulum and Golgi apparatus, are probably a result of hormonal imbalance in lizards of both sexes gonadectomized for one or two months.


Expression and compartmentalization of integral plasma membrane proteins by hepatocytes and their progenitors in the rat pancreas.
A combination of Western blotting, Northern blotting and immunofluorescence was used to examine the expression and compartmentalization of plasma membrane proteins by those hepatocyte-like cells that arise in the pancreases of rats subjected to sequential dietary copper depletion and repletion. The pancreatic hepatocytes were found to: (1) express several integral membrane proteins known to be concentrated within the apical, lateral or basolateral domains of the plasma membranes of hepatocytes in liver; and (2) compartmentalize the membrane proteins to equivalent plasma membrane domains, despite the organization of these cells into clusters instead of highly vascularized plates. The apical plasma membrane proteins dipeptidylpeptidase IV and HA 4 were found to line bile canaliculus-like openings between adjacent pancreatic hepatocytes; these openings were shown to be continuous with the pancreatic exocrine duct by India ink infusion. In contrast, the basolateral plasma membrane protein rat hepatic lectin-1 and lateral plasma membrane protein HA 321 were detected elsewhere about the surfaces of the pancreatic hepatocytes: by analogy to their respective localizations on hepatocytes in liver, rat hepatic lectin-1 was concentrated on those surfaces exposed to the pancreatic matrix at the periphery of the hepatocyte clusters (the basal surface equivalent), whereas HA 321 was concentrated on those surfaces exposed to adjacent hepatocytes within the clusters. The hepatocyte plasma membrane proteins were found to be expressed in the pancreas at different times during the copper depletion/repletion protocol: for example, rat hepatic lectin-1 and the bulk of the HA 4 were expressed relatively late in the protocol, only after large numbers of pancreatic hepatocytes had appeared; whereas dipeptidylpeptidase IV was induced greater than 10-fold early in the protocol and proved to be an apical-specific marker for those ductular epithelial cells that are believed to be the progenitors of the pancreatic hepatocytes.


Folic acid for the prevention of neural tube defects: the Danish experience.
Evidence from controlled trials suggests that ingestion of 0.4 mg of folic acid per day in the periconceptional period is effective in preventing neural tube defects (NTD). For this reason, most countries recommend that women planning pregnancy take folic acid supplements in the periconceptional period, and some countries even fortify stable foods with folic acid. Denmark exemplifies a country with a relatively conservative attitude with respect to taking action in these matters. In 1999, a national information campaign was launched that recommended women planning pregnancy take 0.4 mg of folic acid periconceptionally, but with the moderation that women who eat a healthy diet do not need to take folic acid supplement. The campaign was repeated during 2001. The results of the latter campaign were evaluated by using data from a national survey among pregnant women conducted simultaneously with the campaign by the Danish National Birth Cohort. An increase in the proportion of folic acid users took place concomitantly with the launching of the information events, but the increase was limited. Among women who did not plan their pregnancy, a small proportion had taken folic acid supplements periconceptionally, and this proportion did not change concomitantly with the campaign. Young age and low education were factors associated with low likelihood of taking folic acid. It seems that different and more efficient actions are needed if a more substantial proportion of Danish women and their fetuses are going to benefit from the knowledge that folic acid supplementation in the periconceptional period can prevent NTD.


Mode of infection, nodulation specificity, and indigenous plasmids of 11 fast-growing Rhizobium japonicum strains.
Eleven fast-growing strains of Rhizobium japonicum were characterized with respect to indigenous plasmids and abilities to infect (Inf+) and nodulate (Nod+) cowpea, siratro, wild soybean, and three commercial cultivars of soybean. All strains caused infection via infection threads in root hairs and consistently nodulated cowpea, siratro, and wild soybean in growth pouches. Interactions with commercial cultivars of soybean were strikingly strain specific. Some combinations were Nod-, and infection was delayed in others. The ratios of infections to nodules and the distribution of nodules on primary and lateral roots also varied substantially. A modified in-gel lysis procedure was devised for electrophoretic separation of plasmids from the strains. Plasmids (ranging in size from 35 to greater than 300 megadaltons) were reproducibly detected in all strains.


An emerging global programme against tuberculosis: agenda for research, including the impact of HIV infection.
During the last 5 years, a worldwide resurgence of tuberculosis has been convincingly documented. Increased numbers of cases have been reported from several industrialized countries and there has been a veritable explosion of tuberculosis in the developing countries of sub-Saharan Africa. The fact that most of this increase is attributable to coexisting infection with the human immunodeficiency virus (HIV) should come as no surprise, because of the role played by cell-mediated immunity in protecting against tuberculosis and the depletion of this immunity by HIV infection. The World Health Organization estimates that there are more than 3 million persons in the world dually infected with HIV and tubercle bacilli. To combat this problem, there needs to be an expansion of programmes designed to identify and treat all patients with tuberculosis, whether HIV infected or not. In addition, the prospects for further advances in basic research, clinical research, epidemiological research and operations research, which have broad application to clinical medicine, are great, and these results will help combat the dual scourges of HIV infection and tuberculosis.


Physicians' attitudes towards and reasons for participation in the Candesartan Antihypertensive Survival Evaluation in Japan (CASE-J) trial.
Physicians' perception and attitudes towards a research topic and trial management could influence their participation in a randomized controlled trial. The objectives of this study were to determine the reasons for physicians' participation in and attitudes towards the Candesartan Antihypertensive Survival Evaluation in Japan (CASE-J) trial. CASE-J's main objective is to compare the effectiveness of an angiotensin II receptor antagonist (candesartan cilexetil) with that of a calcium channel blocker (amlodipine besilate) in terms of the incidence of cardiovascular events among high-risk hypertensive patients. We conducted a questionnaire survey among the physicians (n = 512) participating in that trial to determine the reasons behind their participation and to elicit their reactions to the trial management. Eighty-eight percent of the 512 participating physicians responded to our survey. The main reasons for participation were clear objectives of the trial (85.1%), a simple protocol (61.1%), interest in finding out the inhibiting effects of the drugs on cardiac events (80.2%), and a well-organized support system (59.8%). As for negative factors, case registration and follow-up were considered cumbersome by 28.6% and 10.8%, respectively while 44.2% stated that support by the clinical research coordinators provided by the trial management authority was necessary for case screening, recruitment process, patient registration, and follow-up. Multivariate logistic regression analysis showed that participants who did not use a computer very regularly (odds ratio = 1.9, 95% confidence interval = 1.1-3.6) were more likely to consider the case registration and follow-up procedures as a cumbersome. The information generated by this study could be useful in designing future randomized controlled trials in Japan and abroad.


Promoter structure-dependent functioning of the general transcription factor IIE in Saccharomyces cerevisiae.
General transcription factor (TF) IIE is an essential component of the basal transcription complex for protein-encoding genes, which is widely conserved in eukaryotes. Here we analyzed requirement for TFIIE for transcription in vivo by using yeast Saccharomyces cerevisiae cells harboring mutations in the TFA1 gene encoding the larger one of the two subunits of TFIIE. Deletion analysis indicated that the N-terminal half of Tfa1 protein has an essential function to support the cell growth. In a temperature-sensitive tfa1 mutant cell, the steady-state level of bulk poly(A)+ RNA decreased rapidly at the restrictive temperature. Surprisingly, levels of several mRNAs, whose transcription is directed by the promoters lacking the typical TATA sequence, were not affected in the mutant cells at that temperature. This promoter-specific functioning of TFIIE was reproduced in a cell-free system composed of TFIIE-depleted nuclear extracts. These results strongly suggest that requirement for TFIIE varies in each gene depending on the promoter structures in vivo.


Effects of Chronic Intractable Insomnia on Inflammatory Cytokines, Blood Pressure Characteristics, and Antihypertensive Efficacy in Hypertensive Patients.
BACKGROUND This study investigated the changes in blood pressure and inflammatory cytokines in patients with chronic intractable insomnia, and explored the effects of chronic intractable insomnia on antihypertensive efficacy. MATERIAL AND METHODS A total of 248 patients with hypertension admitted to our hospital from 2008 to 2017 were enrolled. We enrolled 124 patients without chronic insomnia in the control group, while 124 patients with chronic insomnia were included in the treatment group. The treatment group received estazolam and was further subdivided into the effective group (n=96) and the ineffective group (n=28) according to Sleep Dysfunction Rating Scale (SDRS) scores. Sleep quality before and after treatment was determined. RESULTS Antihypertensive treatment with eplerenone (50 mg) significantly reduced SDRS scores, C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), matrix metalloproteinase-9 (MMP-9), serum intercellular adhesion molecules (sICAM), and IL-1β levels, as well as systolic blood pressures (SBP) and diastolic blood pressures (DBP), with elevation of non-dipper blood pressure rhythm (P<0.05). The inhibition of intractable insomnia significantly downregulated SBP and DBP, as well as serum inflammatory cytokines such as CRP and TNF-α, showing a favorable effect on antihypertensive function. CONCLUSIONS Alleviation of chronic intractable insomnia facilitates hypertension therapy through decreasing levels of inflammatory cytokines and the proportion of non-dipper blood pressure rhythm, which offers insights for the treatment of hypertension.


Needlestick injuries among medical students: incidence and implications.
To determine the incidence of needlestick injuries in medical school and to examine the behaviors associated with reporting injuries to an occupational health office. Medical students have underdeveloped surgical skills and are at high risk of needlestick injuries. Recent medical school graduates enrolled in a surgery residency at 17 medical centers were surveyed regarding needlestick injuries that they sustained during medical school. The survey asked about the circumstances and cause of injury and postinjury reporting. Of 699 respondents, 415 (59%) reported having sustained a needlestick injury as a medical student; the median number of injuries per injured respondent was 2 (interquartile range: 1-2). Respondents who sustained a needlestick injury in medical school were more likely to sustain a needlestick injury during residency than those who did not experience a needlestick injury in medical school (odds ratio [OR]: 2.57; 95% CI: 1.84, 3.58). Of 89 residents who sustained their most recent needlestick injury during medical school, 42 (47%) did not report their injury to an employee health office. Needlestick injuries and underreporting of these injuries are common among medical students and place them at risk for hepatitis and human immunodeficiency virus. Strategies aimed at improving reporting systems and creating a culture of reporting should be implemented by medical centers.


Older adults' acceptance of a robot for partner dance-based exercise.
Partner dance has been shown to be beneficial for the health of older adults. Robots could potentially facilitate healthy aging by engaging older adults in partner dance-based exercise. However, partner dance involves physical contact between the dancers, and older adults would need to be accepting of partner dancing with a robot. Using methods from the technology acceptance literature, we conducted a study with 16 healthy older adults to investigate their acceptance of robots for partner dance-based exercise. Participants successfully led a human-scale wheeled robot with arms (i.e., a mobile manipulator) in a simple, which we refer to as the Partnered Stepping Task (PST). Participants led the robot by maintaining physical contact and applying forces to the robot's end effectors. According to questionnaires, participants were generally accepting of the robot for partner dance-based exercise, tending to perceive it as useful, easy to use, and enjoyable. Participants tended to perceive the robot as easier to use after performing the PST with it. Through a qualitative data analysis of structured interview data, we also identified facilitators and barriers to acceptance of robots for partner dance-based exercise. Throughout the study, our robot used admittance control to successfully dance with older adults, demonstrating the feasibility of this method. Overall, our results suggest that robots could successfully engage older adults in partner dance-based exercise.


The mechanism of enzymatic cellulose degradation. Purification and some properties of two different 1,4beta-glucan glucanohydrolases from Trichoderma viride.
1. A low-molecular-weight and a high-molecular-weight 1,4beta-glucan glucanohydrolase (Cx enzyme) have been isolated from a commercial cellulase preparation derived from culture filtrates of the fungus Trichoderma viride. 2. The purification method for the isolation of the low-molecular-weight enzyme is a three-step procedure including chromatography on Bio-Gel P-10, chromatography on a dipolar adsorbent (arginine-Sepharose 6 B) and isoelectric focusing. 3. The starting material for the isolation of the high-molecular-weight enzyme was pre-fractionated by chromatography on Bio-Gel P-10, by DEAE-Sephadex chromatography and by SE-Sephadex chromatography as described previously by us. Further fractionation of this material was achieved by affinity chromatography and repeated isoelectric focusing. 4. Free zone electrophoresis of the low-molecular-weight enzyme indicated a homogeneous protein. The high-molecular-weight enzyme was homogenous in sedimentation equilibrium analysis. 5. The molecular weights of the enzymes were 12 500 and 50 000 +/- 2000 respectively. The former value was determined by chromatography on a calibrated column of Bio-Gel P-100 and the latter value by sedimentation equilibrium analysis. 6. The low-molecular-weight enzyme was isoelectric at pH 4.60 (10 degrees C) and contained 21% carbohydrate. The corresponding values for the high-molecular-weight enzyme were pH 3.39 and 12%. 7. Both enzymes were active in releasing free fibers from filter-paper. The low-molecular-weight enzyme was estimated to be about twice as effective as the high-molecular-weight enzyme in this regard.


Synchrotron Infrared Absorbance Measurements of Hydrogen in MgSiO3 Perovskite.
Micro-infrared spectroscopic measurements on single crystals of MgSiO(3) perovskite document two pleochroic hydroxyl absorbance peaks at 3483 and 3423 centimeter(-1). These measurements were obtained with the use of a synchrotron infrared source for spectroscopy. These data are consistent with a trace hydrogen content of 700 +/- 170 hydrogen atoms per 10(6) silicon atoms in the nominally anhydrous MgSiO(3) perovskite. When integrated over the volume of the lower mantle, this concentration is comparable to 12 percent of the mass of hydrogen in the Earth's hydrosphere.


Technetium Nitrido Complexes of Tetradentate Thiosemicarbazones: Kit-Based Radiolabeling, Characterization, and In Vivo Evaluation.
Bis(thiosemicarbazone) and pyridylhydrazone-thiosemicarbazone chelators have demonstrated utility in nuclear medicine. In particular, the 64Cu2+ complexes have been extensively developed for hypoxia imaging and molecular imaging of peptide and protein markers of disease. However, the chemistry and application of bis(thiosemicarbazone) and pyridylhydrazone-thiosemicarbazone chelators in combination with 99mTc, the most widely used radionuclide in nuclear medicine, is underexplored. Herein, a series of bis(thiosemicarbazone) and pyridylhydrazone-thiosemicarbazone chelators were radiolabeled with nitrido-technetium-99m in an optimized one-pot synthesis from [99mTc]TcO4-. Optimization of the radiochemical syntheses allowed for production of the complexes in >90% radiochemical conversion with apparent molar activities of 3.3-5 GBq/μmol. Competition experiments demonstrated the excellent stability of the complexes. The nitrido-technetium-99 complexes were synthesized, and the chemical identities were investigated using mass spectrometry, spectroscopy, and density functional theory calculations. Complexation of nitrido-rhenium(V) was achieved with the N4-dialkylated bis(thiosemicarbazones). Planar imaging and ex vivo biodistribution studies of the five 99mTc complexes were conducted on healthy BALB/c mice to determine in vivo behavior. The lipophilic nature of the complexes resulted in uptake of 1.6-5.7% ID g-1 in the brain at 2 min postinjection and retention of 0.4-1.7% ID g-1 at 15 min postinjection. The stability of the complexes and the biodistribution data demonstrate that these chelators are ideal platforms for future production of radiopharmaceutical candidates.


Carney complex, a familial multiple neoplasia and lentiginosis syndrome. Analysis of 11 kindreds and linkage to the short arm of chromosome 2.
Carney complex is an autosomal dominant syndrome characterized by multiple neoplasias, including myxomas at various sites and endocrine tumors, and lentiginosis. The genetic defect(s) responsible for the complex remain(s) unknown. We studied 101 subjects, including 51 affected members, from 11 North American kindreds with Carney complex. Blood samples were collected from patients and their family members. Hospital records, photographs, and tissue specimens of deceased individuals were reviewed. DNA was extracted from blood samples, patient-derived cell lines, and/or paraffin-embedded tissues. Linkage analysis was performed with highly polymorphic microsatellite markers, distributed over areas of the human genome harboring the most likely candidate genes. The most prevalent clinical manifestation in patients with Carney complex was spotty skin pigmentation, similar to that observed in Peutz-Jeghers and other lentiginosis syndromes. Skin and cardiac myxomas, Cushing syndrome, and acromegaly were present in 62, 30, 31 and 8 percent of the patients, respectively. Linkage was obtained for three markers on the short arm of chromosome 2 (2p16), with a maximum two-point lod score of 5.97 at theta = 0.03 for the marker CA-2 (odds in favor of linkage 10(6):1. The flanking markers CA7 and D2S378 defined a region of approximately 6.4 cM that is likely to contain the gene(s) associated with Carney complex. Candidate genes in the proximity, including the propiomelanocortin and the DNA-mismatch repair hMSH2 genes, were excluded. We conclude that the genetic defect(s) responsible for Carney complex map(s) to the short arm of chromosome 2 (2p16). This region has exhibited cytogenetic aberrations in atrial myxomas associated with the complex, and has been characterized by microsatellite instability in human neoplasias.


Is Anticoagulation Necessary for Severely Disabled Cardioembolic Stroke Survivors?
Background and Objectives: Oral anticoagulants are the hallmark of cardioembolic stroke prevention, but they are frequently underused, especially in elderly patients and patients with paroxysmal atrial fibrillation. In our paper, we analyzed the long-term outcome of severely disabled cardioembolic stroke survivors depending on the prescribed antithrombotic secondary prevention medication. Materials and Methods: In our study, we retrospectively collected data for ischemic stroke (IS) patients treated in P. Stradins Clinical University hospital, Riga, Latvia, from 2014 until 2017. Patients' clinical data were collected using local stroke registry, including patients' demographic data, vascular risk factors, clinical findings, and laboratory results. Severely disabled stroke survivors were followed up by phone at 30/90/180/365 days after discharge. Patients' functional outcomes were assessed using the adapted version of The Rankin Focused Assessment-Ambulation. The collected data were compared in 4 groups according to prescribed secondary prevention medication. Results: A total of 682 (91.42%) patients were followed up and included in data analysis. The median age of patients was 80 (IQR = 75-85) years. Of these patients, 231 (31%) were males and 515 (69%) were females. One-year probability of survival of patients not taking any preventive medication was 53% (IQR = 29-76), while in patients taking antiplatelet agents it was 57% (IQR = 37-78), 78% (IQR = 68-88) of patients on Vitamin K antagonists (VKA) and 81% (IQR = 72-90) in patients on direct oral anticoagulants (DOACs). One year after discharge 73 (31%) had mRS 0-2, 50 (20.9%), 29 (12.1%) were still severely disabled, and 87 (36.4%) had died. Conclusions: Anticoagulant use in secondary prevention predicts better functional outcome and higher survival rate in patients with severe cardioembolic stroke due to non-valvular atrial fibrillation (NVAF), therefore severe neurological deficit must not be a reason of restriction of anticoagulation.


Simultaneous determination of non-volatile, semi-volatile, and volatile organic acids in tobacco by SIM-Scan mode GC-MS.
A method for simultaneous determination of seven nonvolatile organic acids (NVOAs), five semivolatile organic acids (SVOAs), and twelve volatile organic acids (VOAs) in tobacco by synchronous SIM (Selected Ion Monitoring)-scan mode GC-MS was developed. The collection of NVOAs and SVOAs data was performed in full-scan mode, and that of VOAs data was carried out with SIM. Recoveries of NVOAs and SVOAs varied from 90.0 to 103.0%, and RSDs were less than 4.0%; recoveries of VOAs ranged from 89.5 to 99.3%, and their RSDs were less than 3.0%. Twelve tobacco samples were analyzed by the described method, and the results show that the method is applicable for the simultaneous determination of VOAs, NVOAs, and SVOAs in tobacco.


Multi-step dielectrophoresis for separation of particles.
A new concept for separation of particles based on repetitive dielectrophoretic trapping and release in a flow system is proposed. Calculations using the finite element method have been performed to envision the particle behavior and the separation effectiveness of the proposed method. As a model system, polystyrene beads in deionized water and a micro-flow channel with arrays of interdigited electrodes have been used. Results show that the resolution increases as a direct function of the number of trap-and-release steps, and that a difference in size will have a larger influence on the separation than a difference in other dielectrophoretic properties. About 200 trap-and-release steps would be required to separate particles with a size difference of 0.2%. The enhanced separation power of dielectrophoresis with multiple steps could be of great importance, not only for fractionation of particles with small differences in size, but also for measuring changes in surface conductivity, or for separations based on combinations of difference in size and dielectric properties.


Quality of Sleep in the Cypriot Population and Its Association With Multimorbidity: A Cross-Sectional Study.
Poor sleep is a relatively common condition with possibly serious adverse health consequences. Lack of sleep affects the endocrine, immune, and nervous systems. In Cyprus, there is no information about the quality of sleep in the population. The goal of this study was to assess the quality of sleep in the Cypriot population and evaluate its association with multimorbidity. A representative sample of the adult population of Cyprus was selected in 2018-2019 among the five government-controlled municipalities of the Republic of Cyprus using stratified sampling. Data on sleep quality as well as on the presence of chronic, clinical, and mental health conditions were collected using a validated questionnaire. Diseases were classified according to the International Classification of Diseases, 10th Revision (ICD-10). A total of 1,140 Cypriot men and women over 18-years of age (range: 18-94) participated in the study. The median Pittsburgh sleep quality index score of the participants was 5 (first quartile = 3, third quartile = 7) with the maximum score being 17, which suggests that the Cypriot population has a relatively good quality of sleep overall, although, almost one-third of the study population had a poor quality of sleep. Women, residents of Paphos, and married people had a poorer quality of sleep (p < 0.05). Having a poor quality of sleep was associated with higher odds of multimorbidity (OR = 2.21, 95% CI: 1.55, 3.16), even after adjusting for demographics, socioeconomic, and lifestyle factors. Adopting good sleep habits could be beneficial and would potentially help reduce the risk of multimorbidity. Public health guidelines regarding the importance of sleep and its association with multimorbidity should be considered.


The Next Generation of Machine Learning in DDIs Prediction.
Drug-drug interactions may occur when combining two or more drugs may cause some adverse events such as cardiotoxicity, central neurotoxicity, hepatotoxicity, etc. However, a large number of researchers who are proficient in pharmacokinetics and pharmacodynamics have been engaged in drug assays and trying to find out the side effects of all kinds of drug combinations. However, at the same time, the number of new drugs is increasing dramatically, and the drug assay is an expensive and time-consuming process. It is impossible to find all the adverse reactions through drug experiments. Therefore, new attempts have been made in using computational techniques to deal with this problem. In this review, we conduct a review of the literature on applying the computational method for predicting drug-drug interactions. We first briefly introduce the widely used data sets. After that, we elaborate on the existing state-of-art deep learning models for drug-drug interactions prediction. We also discussed the challenges and opportunities of applying the computational method in drug-drug interactions prediction.


[Cortex-wall connections in the apical cell of Sphacelaria].
The apical cell of Sphacelaria (Fucophyceae) exhibits a permanent polarized organization throughout asymmetric divisions. The apex organization was studied by immunolocalization of tubulin, vitronectin, alpha-actinin and beta 1 integrin. Microfilaments were stained directly by fluorescein phalloidin. The apex was highly organized around a patch of microfilaments densely packed at the tip, where vitronectin-like and alpha-actinin-like proteins colocalized. In the same area, an actin-dependent targeted secretion of sulfated polysaccharides was shown. The permanent localization of these components throughout cell elongation suggests that a cortical site involving transmembrane connections between the cytoskeleton and the extracellular matrix is required for cell polarity. A model of the organization of the tip is proposed.


Surface structures and adherence properties of diverse strains of Haemophilus influenzae biogroup aegyptius.
Haemophilus influenzae biogroup aegyptius is an important cause of conjunctivitis and has recently been associated with Brazilian purpuric fever (BPF), a fulminant systemic disease of children. To gain insight into the bacterial factors involved in the pathogenesis of this disease, we investigated the surface structures and adherence properties of eight different strains of H. influenzae biogroup aegyptius, including both BPF and non-BPF isolates. All eight strains were able to express long peritrichous pili similar to those observed in H. influenzae. As in H. influenzae, piliation correlated with colony binding of human erythrocytes. However, two strains were capable of hemagglutination in the absence of pili; in these strains, hemagglutination was insensitive to protease treatment, suggesting a nonproteinaceous hemagglutinin. All strains possessed short, thin, surface fibers distinct from long pili and demonstrated efficient attachment to cultured human conjunctival cells. Attachment to conjunctival cells occurred independently of long pili or a capacity for hemagglutination.


[Endoscopic removal of gastric polyps in ambulatory conditions].
The study included 325 patients (age 16-89 years; 84 males and 241 females) who had undergone surgery for stomach polyps under outpatient hospital conditions. On the whole, 752 polyps were removed: 623 sat on a wide base and 129--on a pedicle. Polyps were removed if bases were under 2.5 cm in diameter and there were no concomitant grave pathologies or blood coagulation disorders. During eight years of follow-up, recurrences were identified in 18, and polyps residing in other parts of the stomach--in 114 patients. Complications which were not dangerous or required surgery occurred in 52 cases. Among the most frequent complications was bleeding which either ceased or was arrested by endoscopic hemostatics. No lethality was recorded. It is concluded that gastric polypectomy in most patients may be performed in outpatient hospitals. If carried out by experienced staff, the procedure will not involve grave complications.


Differential localization profile of Fras1/Frem proteins in epithelial basement membranes of newborn and adult mice.
The Fras1/Frem gene family encodes for structurally similar proteins of the extracellular matrix, functionally correlated with embryonic dermal-epidermal adhesion as deduced from the appearance of sub-epidermal blisters in mouse mutants compromising the function of Fras1, Frem1 and Frem2 proteins. Mutations in the human counterparts FRAS1 and FREM2 have been detected in patients suffering from Fraser syndrome. So far, Fras1/Frem proteins have been shown to be strictly colocalized in the sublamina densa of mouse epithelial basement membranes during development. Here, we focused on the characterization of the localization pattern of the aforementioned proteins, in various parts of the adult mouse skin as well as a range of organs and tissues. Frem3 was present in a broad range of epithelial basement membranes where Fras1, Frem1 and Frem2 were missing. The localization profile of Frem3 coincided with that of collagen VII in all skin basement membranes but differed in that Frem3 was additionally found in the basement membrane of several internal epithelia, where collagen VII was absent. Fras1 and Frem2 were colocalized with Frem3 in the basement membrane of certain skin parts, underlying the thin-layer, of rapidly proliferating keratinocytes, whereas Frem1 was detected only in the basement membrane of the tail. The localization pattern of Fras1 and Frem2 was indistinguishable, while both proteins along with Frem3 could be detected even in the absence of Frem1.


Transmission electron microscopy of interfaces in structural ceramic composites
Ceramic composites based either on a particulate, fibre or a lamellar architecture are potentially useful as damage-tolerant high-temperature engineering materials. The ability of the interfaces in such systems to deflect cracks is vital to the damage tolerance of these materials. Transmission electron microscopy techniques enable the chemical and physical characterization of these interfaces, providing information on interlayer thicknesses, chemical species, local bonding and the microstructural features which give rise to the interfacial properties, thereby enabling a full understanding not only of composites after processing, but also after exposure to aggressive environments such as air at high temperature. Examples of the application of transmission electron microscopy to all three composite architectures are described.


[Applied medicine and ophthalmology].
Medicine as applied by specialists is an unexplored territory. By the example of Ophthalmology some relevant, specific scientific problems are outlined here.


Effects of energy density of daily food intake on long-term energy intake.
An important question that has been raised recently is whether it is mainly the energy density (ED) of the food consumed or its macronutrient composition that determines daily energy intake (EI). In this scope, the effect of ED on EI has been assessed in short-term as well as long-term experiments. Over the short term, i.e., during a meal, it was found that ED affects EI directly; then subjects mainly monitor the weight of the food ingested. Over the long term, the effects of ED on EI are modulated. Average daily energy intake (ADEI) does not only consist of meals but also includes snacks and drinks. ADEI appears to be related to ED of the food and drinks where ED is at least determined by specific macronutrients (primarily fat and carbohydrate), but not when ED is determined only by the weight of water. With respect to the separate effects of the ED of foods and of drinks on ADEI, only ED from foods had a significant relationship with total EI. Moreover, during daily food intake, subjects seem to adapt their portion sizes to estimated EDs. Long-term studies have shown that dietary restraint compensates for the effect of increases in ED on ADEI, whereas unrestrained eaters compensate for the effect of decreases in ED on ADEI. In conclusion, ED determines short-term EI. This cannot be extrapolated to the long term because only the ED of food, and not the ED of drinks, determines total EI. In addition, over the long term, the short-term effect is modulated by dietary restraint and adapted portion sizes. ED is not a universal concept that determines EI, yet rather a characteristic of the macronutrients: mainly fat and carbohydrate contributing to variation in EI.


Correlated matrix-fluctuation-mediated activated transport of dilute penetrants in glass-forming liquids and suspensions.
We formulate a microscopic, force-level statistical mechanical theory for the activated diffusion of dilute penetrants in dense liquids, colloidal suspensions, and glasses. The approach explicitly and self-consistently accounts for coupling between penetrant hopping and matrix dynamic displacements that actively facilitate the hopping event. The key new ideas involve two mechanistically (at a stochastic trajectory level) coupled dynamic free energy functions for the matrix and spherical penetrant particles. A single dynamic coupling parameter quantifies how much the matrix displaces relative to the penetrant when the latter reaches its transition state which is determined via the enforcement of a temporal causality or coincidence condition. The theory is implemented for dilute penetrants smaller than the matrix particles, with or without penetrant-matrix attractive forces. Model calculations reveal a rich dependence of the penetrant diffusion constant and degree of dynamic coupling on size ratio, volume fraction, and attraction strength. In the absence of attractions, a near exponential decrease of penetrant diffusivity with size ratio over an intermediate range is predicted, in contrast to the much steeper, non-exponential variation if one assumes local matrix dynamical fluctuations are not correlated with penetrant motion. For sticky penetrants, the relative and absolute influence of caging versus physical bond formation is studied. The conditions for a dynamic crossover from the case where a time scale separation between penetrant and matrix activated hopping exists to a "slaved" or "constraint release" fully coupled regime are determined. The particle mixture model is mapped to treat experimental thermal systems and applied to make predictions for the diffusivity of water, toluene, methanol, and oxygen in polyvinylacetate liquids and glasses. The theory agrees well with experiment with values of the penetrant-matrix size ratio close to their chemically intuitive values.


Comparison of eye morphology and retinal topography in two species of New World vultures (Aves: Cathartidae).
Vultures are highly reliant on their sensory systems for the rapid detection and localization of carrion before other scavengers can exploit the resource. In this study, we compared eye morphology and retinal topography in two species of New World vultures (Cathartidae), turkey vultures (Cathartes aura), with a highly developed olfactory sense, and black vultures (Coragyps atratus), with a less developed sense of olfaction. We found that eye size relative to body mass was the same in both species, but that black vultures have larger corneas relative to eye size than turkey vultures. However, the overall retinal topography, the total number of cells in the retinal ganglion cell layer, peak and average cell densities, cell soma area frequency distributions, and the theoretical peak anatomical spatial resolving power were the same in both species. This suggests that the visual systems of these two species are similar and that vision plays an equally important role in the biology of both species, despite the apparently greater reliance on olfaction for finding carrion in turkey vultures.


Shiga toxin 2f-producing Escherichia albertii from a symptomatic human.
The previously identified Shiga toxin (Stx) 2f-producing Escherichia coli O115:HNM strain F08/101-31, isolated from a symptomatic human, was confirmed to be E. albertii in the present study by whole genome DNA-DNA hybridizations, by sequencing (cpn60, dnaJ, and 16S rRNA genes), and by multi-locus sequence typing. The F08/101-31 strain was originally identified as E. coli rather than the relatively new bacterial species E. albertii, which was first described in 2003, because it did not display any of the biochemical characteristics of E. albertii. This new classification will impact public health management strategies in Japan because the present study showed that some E. albertii strains, which are often misidentified as E. coli, produce Stx and likely cause diarrhea in humans. Therefore, further guidelines for the management and identification of Stx-producing E. albertii are required in Japan.


Metal-Organic Frameworks in Mixed-Matrix Membranes for High-Speed Visible-Light Communication.
Mixed-matrix membranes (MMMs) based on luminescent metal-organic frameworks (MOFs) and emissive polymers with the combination of their unique advantages have great potential in separation science, sensing, and light-harvesting applications. Here, we demonstrate MMMs for the field of high-speed visible-light communication (VLC) using a very efficient energy transfer strategy at the interface between a MOF and an emissive polymer. Our steady-state and ultrafast time-resolved experiments, supported by high-level density functional theory calculations, revealed that efficient and ultrafast energy transfer from the luminescent MOF to the luminescent polymer can be achieved. The resultant MMMs exhibited an excellent modulation bandwidth of around 80 MHz, which is higher than those of most well-established color-converting phosphors commonly used for optical wireless communication. Interestingly, we found that the efficient energy transfer further improved the light communication data rate from 132 Mb/s of the pure polymer to 215 Mb/s of MMMs. This finding not only showcases the promise of the MMMs for high-speed VLC but also highlights the importance of an efficient and ultrafast energy transfer strategy for the advancement of data rates of optical wireless communication.


Thoracic Outlet Syndrome Caused by a Primary Tumour in the Brachial Plexus.
Thoracic outlet syndrome (TOS) caused by a primary brachial plexus tumour is very rare. A male politician in his 40s presented with numbness, left limb pain and positive Wright and Roos test results. Magnetic resonance imaging (MRI) revealed a tumour located just below the clavicle, compressing the subclavian artery during left arm elevation. Despite concerns regarding postoperative nerve deficits, surgery was performed because of worsening symptoms during the election campaigns. The pathology report revealed a schwannoma. Few reports have described TOS caused by primary tumours of the brachial plexus. While the decision to perform surgery for primary tumours of the brachial plexus requires careful consideration, surgery may be indicated in cases where the tumour location causes such symptoms. Level of Evidence: Level V (Therapeutic).


Looking at the (w)hole: magnet resonance imaging in polycystic kidney disease.
Inherited cystic kidney diseases, including autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD), are the most common monogenetic causes of end-stage renal disease (ESRD) in children and adults. While ARPKD is a rare and usually severe pediatric disease, the more common ADPKD typically shows a slowly progressive course leading to ESRD in adulthood. At the present time there is no established disease-modifying treatment for either ARPKD or ADPKD. Various therapeutic approaches are currently under investigation, such as V2 receptor antagonists, somatostatins, and mTOR inhibitors. Renal function remains stable for decades in ADPKD, and thus clinically meaningful surrogate markers to assess therapeutic efficacy are needed. Various studies have pointed out that total kidney volume (TKV) is a potential surrogate parameter for disease severity in ADPKD. Recent trials have therefore measured TKV by magnet resonance imaging (MRI) to monitor and to predict disease progression. Here, we discuss novel insights on polycystic kidney disease (PKD), the value of MRI, and the measurement of TKV in the diagnosis and follow-up of PKD, as well as novel emerging therapeutic strategies for ADPKD.


Effect of phototherapy and exchange transfusion on primary bile acids in the serum of hyperbilirubinaemic newborns.
Serum primary bile acid (cholic (CA) and chenodeoxycholic (CDCA) acid) concentrations were measured in 14 preterm and 11 full-term hyperbilirubinaemic newborns at the beginning and end of, and 24 and 72 hours following phototherapy. Only in the preterm newborns with gestational ages of 35-38 weeks there was a significant decrease of mean serum bile acid concentrations which could be shown 72 hours after the beginning of phototherapy. It can be hypothesized that the decrease was a result of a direct effect of light on the excretory liver function. Serum CA and CDCA concentrations were also measured in 5 hyperbilirubinaemic newborns at the beginning and end, and 24, 48 and 72 hours after the end of exchange transfusion. Exchange transfusion caused a clear immediate decrease in the mean serum primary bile acid concentrations. However, on day 2 after exchange transfusion the mean serum concentration of CA was about 150% and that of CDCA about 110% of the initial values. The most hyperbilirubinaemic newborns had extremely high primary bile acid serum concentrations before therapy. As bile acids compete with bilirubin for albumin binding it should be considered whether high bile acids in the serum of hyperbilirubinaemic newborns presuppose exchange transfusions.


Quantitation of Fas and Fas ligand gene expression in human ovarian, cervical and endometrial carcinomas using real-time quantitative RT-PCR.
Alterations in the expression of Fas (CD95/APO-1) and its ligand (FasL) have been demonstrated in various types of cancers as a mechanism for tumour cell to escape from the immune system. In the present study, we evaluated the expression of the Fas and FasL genes in a wide range of primary gynaecological carcinomas. These included 31 ovarian, 29 cervical and 25 endometrial carcinoma tissues as well as four ovarian and three cervical carcinoma cell lines. Our real-time quantitative reverse transcription polymerase chain reaction analysis revealed that down-regulation of Fas expression is more prominent than the up-regulation of FasL expression in all types of gynaecological cancer studied. This down-regulation of Fas expression was also true for the seven carcinoma cell lines. Only one cervical carcinoma cell line, DoT, exhibited a high level of FasL expression. These results indicated that down-regulation of Fas expression is a common abnormality in many types of cancers including gynaecological cancers, whereas an increase in FasL expression is not a common phenomenon in these cancers.


The efficacy of silver diamine fluoride in arresting caries in children.
Data sourcesPubMed, Scopus, Web of Science, the Latin American and Caribbean Health Sciences Literature database (LILACS), the Brazilian Library in Dentistry (BBO), Cochrane Library and grey literature.Study selectionTwo reviewers selected randomised clinical trials (RCTs) that compared the efficacy of SDF application with other active treatments or placebo in arresting carious lesions.Data extraction and synthesisThree authors extracted data using customised extraction forms, and risk of bias was assessed by two independent reviewers. Meta-analyses were performed on studies classified at 'low' or 'unclear' risk of bias, where similar outcomes were recorded in primary teeth, and that compared SDF to active treatments.ResultsEleven studies were included; five studies were at 'low', two at 'unclear' and four studies at 'high' risk of bias. Eight were conducted with primary teeth, two with permanent first molars and one conducted on both. Six studies used 38% SDF, two 30% SDF, one 12% SDF, one compared 38% SDF to 12% SDF and one used Nano Silver Fluoride (NSF).ConclusionsSDF is more effective than active treatments or placebo for carious lesion arrest in primary teeth. The body of evidence was of high quality for primary teeth. However, there was not enough high quality evidence to draw conclusions about carious lesion arrest in first permanent molars.


[A genetic and physiologic study of mutations of T4 phage suppressing the lysis defect of gene stII mutants].
A new gene rVI phage T4B is found. Mutations rVI belong to the class of r-mutations of T4B. Mutations rVI do not show the lysis inhibition after superinfection and determine the r-morphology of plaques on Escherichia coli CR63 lawn. Mutations rVI are gene-specific suppressors of stII lysis defect, but do not suppress the e lysis defect. Alleles rVI and rVI+ are co-dominated with regard to the ability to suppress the stII lysis defect. The suggest the stoichiometric character of rVI gene function. Mutations rVI increase the permeability of infected cells before the lysis. The gene rVI located between the genes 56 and 39 on the T4B genetic map, close to the gene 56. The still mutations described earlier (Krylov, Yankovsky, 1973) suppress the lysis defect of the am z25 mutant in the gene e. The suppression is manifested when infected bacteria develope only in the liquid media.


Common variation in EMSY and risk of breast and ovarian cancer: a case-control study using HapMap tagging SNPs.
EMSY could be involved in low-level susceptibility to breast and ovarian cancer. Gene amplification is seen in a proportion of breast and ovarian tumours and correlates with poor prognosis in breast cancer patients. Furthermore, the EMSY protein silences a transcription activation domain in BRCA2 exon 3. We used a genetic association study design to determine if common genetic variation (frequency > or = 5%) in EMSY was associated with breast or ovarian cancer risk in the British population. Haplotype tagging single-nucleotide polymorphisms (htSNPs) were selected from the HapMap database and genotyped using Taqman in two large study sets of white British women (n [breast set] = 2343 cases and 2284 controls, n [ovarian set] = 864 cases and 864 controls). HapMap data might be insufficient to tag genetic variation in EMSY comprehensively. We therefore screened the gene promoter and coding sequences with denaturing high performance liquid chromatography in order to identify additional SNPs that are most likely to be functional. HapMap data on 22 SNPs show that 4 htSNPs tag 4 common haplotypes: rs2282611 (5'up t > g), rs4245443 (IVS7 g > a), rs2513511 (IVS16 a > g), rs2155220 (3'down c > t). We observed no association between any of the genotypes or associated haplotypes and breast or ovarian cancer risk. Seventeen out of the 18 remaining HapMap polymorphisms (94%) were well tagged by the 4 selected htSNPs (r2s > 0.8). Genotype frequencies for two further SNPs identified by screening and located near exon-intron boundaries, rs2508740 (IVS9 a > g) and rs11600501 (IVS10 c > t), were also similar in cases and controls. In order to simulate unidentified SNPs, we performed the leave-one-out cross-validation procedure on the HapMap data; over 95% of the common genetic variation was well represented by tagging polymorphisms. We are therefore likely to have tagged any common, functional variants present in our population. We found no association between common genetic variation in EMSY and risk of breast or ovarian cancer in two large study sets of white British women.


Oxygen levels in the gut lumens of herbivorous insects.
Oxygen levels were measured in the foregut and midgut lumens of ten species of caterpillars and three species of grasshoppers. In most species, the foregut was nearly anoxic, with oxygen levels ranging from 0 to 2.5 mm Hg. However, two caterpillar species with large foreguts (Malacosoma disstria and Lymantria dispar) had elevated oxygen levels (27.9 and 32.1 mm Hg) in this region when they were fed artificial diet. In all of the species surveyed, the anterior and posterior midgut were nearly anoxic, with oxygen levels ranging from 0 to 7.3 mm Hg. Oxygen levels in the midgut lumen of Helicoverpa zea did not differ when caterpillars were fed artificial diet or tomato foliage, suggesting that the insect is capable of reducing the level of ingested oxygen in its gut. An examination of the radial microgradient of oxygen in the gut lumen demonstrated that the midgut epithelium is not a sink for ingested oxygen. However, the midgut contents of larvae fed artificial diet were capable of depleting oxygen. This capacity was reduced by boiling, suggesting that the nearly anoxic state of the midgut lumen in some insects is maintained by endogenous chemical processes. We conclude that low oxygen levels in the gut lumens of most herbivorous insects may greatly reduce the rates of oxidation of ingested plant compounds by oxygen-dependent reactions.


Prevalence of syn nucleobases in the active sites of functional RNAs.
Biological RNAs, like their DNA counterparts, contain helical stretches, which have standard Watson-Crick base pairs in the anti conformation. Most functional RNAs also adopt geometries with far greater complexity such as bulges, loops, and multihelical junctions. Occasionally, nucleobases in these regions populate the syn conformation wherein the base resides close to or over the ribose sugar, which leads to a more compact state. The importance of the syn conformation to RNA function is largely unknown. In this study, we analyze 51 RNAs with tertiary structure, including aptamers, riboswitches, ribozymes, and ribosomal RNAs, for number, location, and properties of syn nucleobases. These RNAs represent the set of nonoverlapping, moderate- to high-resolution structures available at present. We find that syn nucleobases are much more common among purines than pyrimidines, and that they favor C2'-endo-like conformations especially among those nucleobases in the intermediate syn conformation. Strikingly, most syn nucleobases participate in tertiary stacking and base-pairing interactions: Inspection of RNA structures revealed that the majority of the syn nucleobases are in regions assigned to function, with many syn nucleobases interacting directly with a ligand or ribozyme active site. These observations suggest that judicious placement of conformationally restricted nucleotides biased into the syn conformation could enhance RNA folding and catalysis. Such changes could also be useful for locking RNAs into functionally competent folds for use in X-ray crystallography and NMR.


Characterization of an in vitro model of elastic fiber assembly.
Elastic fibers consist of two morphologically distinct components: elastin and 10-nm fibrillin-containing microfibrils. During development, the microfibrils form bundles that appear to act as a scaffold for the deposition, orientation, and assembly of tropoelastin monomers into an insoluble elastic fiber. Although microfibrils can assemble independent of elastin, tropoelastin monomers do not assemble without the presence of microfibrils. In the present study, immortalized ciliary body pigmented epithelial (PE) cells were investigated for their potential to serve as a cell culture model for elastic fiber assembly. Northern analysis showed that the PE cells express microfibril proteins but do not express tropoelastin. Immunofluorescence staining and electron microscopy confirmed that the microfibril proteins produced by the PE cells assemble into intact microfibrils. When the PE cells were transfected with a mammalian expression vector containing a bovine tropoelastin cDNA, the cells were found to express and secrete tropoelastin. Immunofluorescence and electron microscopic examination of the transfected PE cells showed the presence of elastic fibers in the matrix. Biochemical analysis of this matrix showed the presence of cross-links that are unique to mature insoluble elastin. Together, these results indicate that the PE cells provide a unique, stable in vitro system in which to study elastic fiber assembly.


Effect of Superior Ovarian Nerve and Plexus Nerve Sympathetic Denervation on Ovarian-Derived Infertility Provoked by Estradiol Exposure to Rats.
Sympathetic innervation of the ovary in rodents occurs via two routes: the superior ovarian nerve (SON), which runs along the ovarian ligament, and the plexus nerve (PN), which is mainly associated with the vasculature. SON and ovarian norepinephrine (NE) levels play a major role in regulating ovarian cystic health. Although it was previously described that the polycystic ovarian phenotype (PCO) is causally related to hyperstimulation of the sympathetic nerves of the ovary, much less is known, however, regarding the role of PN in ovarian physiology. We studied the role of SON and PN in relation to the maintenance of the PCO phenotype induced in the rat by exposure to estradiol valerate (EV). EV exposure at 24 days old (juvenile exposure) increases NE in the ovary for up to 90 days after EV injection. SON or PN denervation (SONX and PNX) decreased NE. SONXreversed the acyclic condition from 30 days after surgery (p < 0.05), but PNXdid not. SONX was more effective than PNX to downregulate the increased number of cysts induced by EV, with the presence of the corpora lutea (CL, signifying ovulation) in the SONX group. Seventy percent of SONX rats presented with pregnancy at 60 days post-EV (6 of the 7 sperm-positive rats were pregnant); however, SONX rats had a reduced number (half) of pups compared with vehicle-treated rats and 60% more pups than EV rats. These data suggest that the SON plays a predominant role in follicular development, ovulation and pregnancy during ovarian diseases.


Network Measures of Collaborative Support for Young Adults With Autism.
The treatment of autism requires complex, multimodal interventions, provided by parents and providers across settings. As young adults with autism spectrum disorder (ASD) transition into adulthood, new stakeholders are added to support planning for postsecondary employment. In this article, we investigate the use of dynamic social network measures that describe the resources of young adults throughout transition. A longitudinal, dynamic social network survey (Social Dynamics of Intervention [SoDI]) was conceptualized, designed, and conducted as a feasibility test, measuring changes in collaboration among parents and school staff members who provided interventions for children with ASD in 2 urban public schools. Using the SoDI, we tracked the following over time: the team who provided interventions, their locations, the interventions they provided, the autism trainings they attended, and the density of the team's problem-solving network for the child. Using the SoDI, we successfully identified stakeholders across settings who provided interventions for each child. Results indicated variation in the density of problem-solving and trust related to ASD intervention across teams as well as variation in intervention and autism knowledge networks during the school year. Adaptations of the SoDI for mapping pre- and posttransition resources and social connectivity across stakeholders for young adults with ASD are proposed in the Discussion section. Dynamic social network approaches can be used to capture changes in intervention, autism knowledge, and social connectivity, providing informative descriptive data about how vocational rehabilitation policies might increase employment supports during the transition process for young adults with ASD.


Prospective, randomized study of atrioventricular ablation and mode-switching, dual chamber pacemaker implantation versus medical therapy in drug-resistant paroxysmal atrial fibrillation. The PAF study. Paroxysmal Atrial Fibrillation.
We performed a prospective randomized 6-month evaluation of the clinical effects of atrioventricular junctional ablation together with placement of a DDDR mode-switching pacemaker vs pharmacological treatment in 43 patients with intolerable paroxysmal atrial fibrillation not controlled with antiarrhythmic drugs. Ablation and pacemaker treatment were highly effective and superior to drug therapy in controlling symptoms and improving quality of life. However, discontinuation of drug therapy exposed patients to further recurrences of paroxysmal atrial fibrillation and the risk of developing permanent atrial fibrillation.


Low molecular weight heparins: structural differentiation by bidimensional nuclear magnetic resonance spectroscopy.
Individual low molecular weight heparins (LMWHs) exhibit distinct pharmacological and biochemical profiles because of manufacturing differences. Correlation of biological properties with particular structural motifs is a major challenge in the design of new LMWHs as well as in the development of generic versions of proprietary LMWHs. Two-dimensional nuclear magnetic resonance (NMR) spectroscopy permits identification and quantification of structural peculiarities of LMWH preparations. In this article, heteronuclear single quantum coherence spectroscopy, previously used to determine variously substituted monosaccharide components of heparan sulfate (HS) and HS-like glycosaminoglycan mimics, has been applied to the structural characterization of three commercially available LMWHs (enoxaparin, dalteparin, and tinzaparin). Relevant residues belonging to the parent heparin, as well as minor residues generated by each depolymerization procedure, have been characterized and quantified. The use of a high-sensitivity NMR spectrometer (600 MHz equipped with cryoprobe) allowed the accurate quantification of residues with sensitivity better than 1 to 2%.


Sustained transgene expression via citric acid-based polyester elastomers.
Polymeric scaffolds are an important tool in tissue engineering and gene delivery using porous scaffolds can be a viable approach to control tissue response. Herein we describe the use of a biodegradable polyester elastomer, poly(1,8-octanediol-co-citrate) (POC), as a substrate for plasmid immobilization and cellular transfection of colonizing cells. Plasmid (pDNA), either complexed with poly(ethyleneimine) (PEI) forming polyplexes or in its native state, was surface-immobilized onto POC scaffolds via adsorption. Polyplex-containing scaffolds showed higher loading and slower initial rates of release than naked pDNA-containing scaffolds. Seeding of HEK293 cells and porcine aortic smooth muscle cells (PASMC) onto polyplex loaded-scaffolds demonstrated cell proliferation and transfection in vitro up to 12 days, significantly longer relative to bolus transfection. In vivo, transfection was evaluated using the mouse intraperitoneal (IP) fat model. In contrast to the in vitro study, successful long-term transgene delivery was only achieved with the naked pDNA-containing scaffolds. In particular, naked pDNA-containing scaffolds promoted high levels of both luciferase and green fluorescent protein (GFP) expression in vivo for 2 weeks. The results demonstrate that POC scaffolds are a suitable material for substrate-mediated gene delivery. POC scaffolds can potentially support long-term biological cues to mediate tissue formation through non-viral gene delivery.


Formylglycine-Generating Enzyme-Like Proteins Constitute a Novel Family of Widespread Type VI Secretion System Immunity Proteins.
Competition is a critical aspect of bacterial life, as it enables niche establishment and facilitates the acquisition of essential nutrients. Warfare between Gram-negative bacteria is largely mediated by the type VI secretion system (T6SS), a dynamic nanoweapon that delivers toxic effector proteins from an attacking cell to adjacent bacteria in a contact-dependent manner. Effector-encoding bacteria prevent self-intoxication and kin cell killing by the expression of immunity proteins, which neutralize effector toxicity by specifically binding their cognate effector and either occluding its active site or preventing the structural rearrangements necessary for effector activation. In this study, we investigate Tsi3, a previously uncharacterized T6SS immunity protein present in multiple strains of the human pathogen Acinetobacter baumannii. We show that Tsi3 is the cognate immunity protein of an antibacterial effector of unknown function, Tse3. Our bioinformatic analyses indicate that Tsi3 homologs are widespread among Gram-negative bacteria, often encoded within T6SS effector-immunity modules. Surprisingly, we found that Tsi3 homologs are predicted to possess a characteristic formylglycine-generating enzyme (FGE) domain, which is present in various enzymatic proteins. Our data show that Tsi3-mediated immunity is dependent on Tse3-Tsi3 protein-protein interactions and that Tsi3 homologs from various bacteria do not provide immunity against nonkin Tse3. Thus, we conclude that Tsi3 homologs are unlikely to be functional enzymes. Collectively, our work identifies FGE domain-containing proteins as important mediators of immunity against T6SS attacks and indicates that the FGE domain can be coopted as a scaffold in multiple proteins to carry out diverse functions. IMPORTANCE Despite the wealth of knowledge on the diversity of biochemical activities carried out by T6SS effectors, comparably little is known about the various strategies that bacteria employ to prevent susceptibility to T6SS-dependent bacterial killing. Our work establishes a novel family of T6SS immunity proteins with a characteristic FGE domain. This domain is present in enzymatic proteins with various catalytic activities. Our characterization of Tsi3 expands the known functions carried out by FGE-like proteins to include defense during T6SS-mediated bacterial warfare. Moreover, it highlights the evolution of FGE domain-containing proteins to carry out diverse biological functions.


[Efficacy of the lithium compound with prolonged action during "reserpine depression"].
A single administration of the new lithium compound with prolonged action formed on the basis of lithium oxyburate produces an antiblepharoptosis effect in rats with reserpine depression for 24 to 48 h. It optimally prevents the development of "reserpine-desynchronization" in rats when prescribed in the morning at intervals of 48 h.


Trends in cardiovascular outcomes after acute coronary syndrome in New Zealand 2006-2016.
Characterisation of trends in acute coronary syndrome (ACS) outcomes are critical to informing clinical practice and quality improvement, but there are few recent population studies for ACS. We reviewed the recent trends in the outcomes of ACS in New Zealand (NZ). All patients with ACS admitted to NZ public hospitals in 2006-2016 were identified from hospital discharge records, and their first ACS hospitalisations per year extracted for analysis. Thirty-day and 1-year death, myocardial infarction, stroke, heart failure and bleeding rates were calculated for each calendar year. Trends in outcome rates were assessed using generalised linear mixed models. Total annual ACS hospitalisations decreased from 685 to 424 per 100 000. Using first patient hospitalisations per year (n=1 55 060), we found significant annual declines in all major outcomes except for non-cardiovascular deaths. All-cause mortality fell from 10.5% to 9.1% at 30 days (adjusted OR 0.985 per year change, p<0.001) and from 21.8% to 18.7% at 1 year (OR=0.994, p=0.016). This was related to significant decreases in cardiovascular death at both time points (OR=0.982 and 0.987, respectively, p<0.001), outweighing a slight increase in non-cardiovascular death at 1 year (OR=1.009, p=0.014). One-year rates of myocardial infarction, heart failure, stroke and bleeding rates all decreased significantly over time. ACS outcomes including all-cause mortality, cardiovascular death, myocardial infarction, stroke, heart failure and bleeding at 30 days and 1 year improved over the last decade in NZ, reflecting successful implementation and advances in prevention, medical and invasive management in ACS over time.


PD-1 inhibitors combined with paclitaxel and cisplatin in first-line treatment of esophageal squamous cell carcinoma (ESCC): a network meta-analysis.
The combinations of PD-1 inhibitors with paclitaxel/cisplatinum (PD-1 + TP) and fluoropyrimidine/cisplatinum (PD-1 + FP) both have been shown to improve overall survival (OS) and progression-free survival (PFS) in patients with previously untreated, advanced esophageal squamous cell carcinoma (ESCC). However, there is no consensus on which chemotherapy regimen combined with PD-1 has better efficacy. To deal with this important issue in the first-line treatment of patients with ESCC, a network meta-analysis (NMA) was performed. Data were collected from eligible studies searched in Medline, Web of Science, PubMed, the Cochrane Library and Embase. The pooled hazard ratio (HR) for the OS, and PFS, odds ratio (OR) for the objective response rate (ORR) and ≥ 3 grade treatment-related adverse events (≥ 3TRAEs) were estimated to evaluate the efficacy of PD-1 inhibitors combined with TP or FP. Five RCTs and one retrospective study involving 3685 patients and evaluating four treatments were included in this NMA. Compared to other treatments, PD-1 + TP was better. For the PFS, the HRs for PD-1 + TP compared to PD-1 + FP, TP and FP were 0.59 (0.44, 0.80), 0.56 (0.51, 0.61) and 0.45 (0.37, 0.56) respectively. For the OS, PD-1 + TP was also a better treatment compared to other treatments. The HRs were 0.74 (0.56, 0.96), 0.64 (0.57, 0.71), 0.53 (0.43, 0.67) respectively. For the ORR, there was no significant difference between PD-1 + TP and PD-1 + FP, and the ORs were 1.2 (0.69, 2.11). Compare with TP and FP, PD-1 + TP had an obvious advantage, ORs were 2.5 (2.04, 3.04) and 2.95 (1.91, 4.63). For ≥ 3TRAEs, PD-1 + TP compared to other treatments, ORs were 1.34 (0.74, 2.46) and 1.13 (0.92, 1.38) and 2.23 (1.35, 3.69). PD-1 + TP significantly improved both PFS and OS compared to PD-1 + FP. Taking into account both efficacy and safety, PD-1 + TP may be a superior first-line treatment option for ESCC.


Toward implantable devices for angle-sensitive, lens-less, multifluorescent, single-photon lifetime imaging in the brain using Fabry-Perot and absorptive color filters.
Implantable image sensors have the potential to revolutionize neuroscience. Due to their small form factor requirements; however, conventional filters and optics cannot be implemented. These limitations obstruct high-resolution imaging of large neural densities. Recent advances in angle-sensitive image sensors and single-photon avalanche diodes have provided a path toward ultrathin lens-less fluorescence imaging, enabling plenoptic sensing by extending sensing capabilities to include photon arrival time and incident angle, thereby providing the opportunity for separability of fluorescence point sources within the context of light-field microscopy (LFM). However, the addition of spectral sensitivity to angle-sensitive LFM reduces imager resolution because each wavelength requires a separate pixel subset. Here, we present a 1024-pixel, 50 µm thick implantable shank-based neural imager with color-filter-grating-based angle-sensitive pixels. This angular-spectral sensitive front end combines a metal-insulator-metal (MIM) Fabry-Perot color filter and diffractive optics to produce the measurement of orthogonal light-field information from two distinct colors within a single photodetector. The result is the ability to add independent color sensing to LFM while doubling the effective pixel density. The implantable imager combines angular-spectral and temporal information to demix and localize multispectral fluorescent targets. In this initial prototype, this is demonstrated with 45 μm diameter fluorescently labeled beads in scattering medium. Fluorescent lifetime imaging is exploited to further aid source separation, in addition to detecting pH through lifetime changes in fluorescent dyes. While these initial fluorescent targets are considerably brighter than fluorescently labeled neurons, further improvements will allow the application of these techniques to in-vivo multifluorescent structural and functional neural imaging.


Functional gene clustering via gene annotation sentences, MeSH and GO keywords from biomedical literature.
Gene function annotation remains a key challenge in modern biology. This is especially true for high-throughput techniques such as gene expression experiments. Vital information about genes is available electronically from biomedical literature in the form of full texts and abstracts. In addition, various publicly available databases (such as GenBank, Gene Ontology and Entrez) provide access to gene-related information at different levels of biological organization, granularity and data format. This information is being used to assess and interpret the results from high-throughput experiments. To improve keyword extraction for annotational clustering and other types of analyses, we have developed a novel text mining approach, which is based on keywords identified at the level of gene annotation sentences (in particular sentences characterizing biological function) instead of entire abstracts. Further, to improve the expressiveness and usefulness of gene annotation terms, we investigated the combination of sentence-level keywords with terms from the Medical Subject Headings (MeSH) and Gene Ontology (GO) resources. We find that sentence-level keywords combined with MeSH terms outperforms the typical 'baseline' set-up (term frequencies at the level of abstracts) by a significant margin, whereas the addition of GO terms improves matters only marginally. We validated our approach on the basis of a manually annotated corpus of 200 abstracts generated on the basis of 2 cancer categories and 10 genes per category. We applied the method in the context of three sets of differentially expressed genes obtained from pediatric brain tumor samples. This analysis suggests novel interpretations of discovered gene expression patterns.


Needs Based Assessment of Trauma Systems 2, is it ready for primetime? A natural experiment testing its reliability.
Needs Based Assessment of Trauma Systems 2 (NBATS-2) attempts to predict the impact on patient volume and travel time for patients when a new trauma center (TC) is added to the system. The purpose of this study was to examine NBATS-2 predictive accuracy regarding expected volume and travel times of trauma patients at a newly designated TC and nearby legacy TCs when compared with actual data. Needs Based Assessment of Trauma Systems predictive model for volume of trauma patients at the new TC was run based on 25th, 50th, and 75th percentiles of both state and National Trauma Data Bank (NTDB) patients per 100 TC beds. This was compared with the actual number of trauma patients from the State Discharge Data set before (2011-2012) and after (2016-2017) designation of the TC. Analysis was then augmented using the geographic information system (ArcGIS) spatial modeling to characterize median travel times for actual trauma patients, before and after designation of the TC. Both state and NTDB 25th, 50th, and 75th percentiles resulted in significant overestimation of volume at the new TC in 2016. After another year of TC maturation (2017), overestimation decreased but was still present. The 25th percentile from state and NTDB data sets provided the most accurate predictions. For the legacy TCs, the model switched from under to overestimation as the state and NTDB percentiles increased. The geographic information system accurately showed patients traveling <40 minutes to a TC nearly doubled. Needs Based Assessment of Trauma Systems 2 provides an excellent template for state strategic planning; however, it overestimates new TC volume and under/overestimates volumes for legacy TCs depending on the state and NTDB percentiles used. This study shows that population density of the county in which the new or legacy TC is located should be considered when choosing the appropriate state or NTDB percentile. The geographic information system appropriately showed a decrease in trauma patient travel times after TC designation. Care Management, level V.


Measurement of in vitro leucocyte mitogenesis in fish: ELISA based detection of the thymidine analogue 5'-bromo-2'-deoxyuridine.
In this study we present a method for the measurement of in vitro mitogenesis in fish leucocytes that is based on the incorporation of the thymidine analogue 5'-bromo-2'-deoxyuridine (BrdU) into the DNA of replicating cells, followed by ELISA-based detection. This technique, adapted from methods developed for mammalian cells, operates on a similar biological principle to (3)H-thymidine incorporation, but circumvents the logistical and safety issues inherent with the radioactive label. Because it directly measures DNA proliferation, the assay has advantages over other colorimetric methods that may be strongly influenced by leucocyte metabolic status. Using BrdU incorporation followed by ELISA, we evaluate the responsiveness of rainbow trout (Oncorhynchus mykiss [Walbaum]) leucocytes to the mammalian T-cell mitogen Concanavalin A (Con A) as well as the differential response of white perch (Morone americana [Gmelin]) leucocytes to Con A and pokeweed mitogen. Specific considerations intrinsic to the assay system are discussed, including the implications of utilising enzyme-based detection.


Predicting Acute Respiratory Distress Syndrome in Severe Blunt Trauma: The Utility of Interleukin-18.
Background: In trauma, direct pulmonary injury and innate immune response activation primes the lungs for acute respiratory distress syndrome (ARDS). The inflammasome-dependent release of interleukin-18 (IL-18) was recently identified as a key mediator in ARDS pathogenesis, leading us to hypothesize that plasma IL-18 is a diagnostic predictor of ARDS in severe blunt trauma. Patients and Methods: Secondary analysis of the Inflammation and Host Response to Injury database was performed on plasma cytokines collected within 12 hours of severe blunt trauma. Trauma-related cytokines, including IL-18, were compared between patients with and without ARDS and were evaluated for association with ARDS using regression analysis. Threshold cytokine concentrations predictive of ARDS were determined using receiver-operating curve (ROC) analysis. Results: Cytokine analysis of patients without ARDS patients (n = 61) compared with patients with ARDS (n = 19) demonstrated elevated plasma IL-18 concentration in ARDS and IL-18 remained correlated with ARDS on logistic regression after confounder adjustment (p = 0.008). Additionally, ROC analysis revealed IL-18 as a strong ARDS predictor (area under the curve [AUC] = 0.83), with a threshold IL-18 value of 170 pg/mL (Youden index, 0.3). Unlike in patients without ARDS, elevated IL-18 persisted in patients with ARDS during the acute injury phase (p ≤ 0.02). Other trauma-related cytokines did not correlate with ARDS. Conclusions: In severe blunt trauma, IL-18 is a robust predictor of ARDS and remains elevated throughout the acute injury phase. These findings support the use of IL-18 as a key ARDS biomarker, promoting early identification of trauma patients at greater risk of developing ARDS. Timely recognition of ARDS and implementation of advantageous supportive care practices may reduce trauma-related ARDS morbidity and costs.


High-performance nanostructured MR contrast probes.
Magnetic resonance imaging (MRI) has become a powerful technique in biological molecular imaging and clinical diagnosis. With the rapid progress in nanoscale science and technology, nanostructure-based MR contrast agents are undergoing rapid development. This is in part due to the tuneable magnetic and cellular uptake properties, large surface area for conjugation and favourable biodistribution. In this review, we describe our recent progress in the development of high-performance nanostructured MR contrast agents. Specifically, we report on Gd-enriched nanostructured probes that exhibit T(1) MR contrast and superparamagnetic Fe(3)O(4) and CoFe(2)O(4) nanostructures that display T(2) MR contrast enhancement. The effects of nanostructure size, shape, assembly and surface modification on relaxivity are described. The potential of these contrast agents for in vitro and in vivo MR imaging with respect to colloidal stability under physiological conditions, biocompatibility, and surface functionality are also evaluated.


[Circulating immune complexes in the pathogenesis of recurrent erysipelas].
The dynamics of circulating immune complexes (CIC) in comparison with the level of SH-groups of serum deproteinate and other characteristics of cell-mediated and humoral immunity (the reaction of the inhibition of antibodies, the levels of T-cells and their main subpopulations) was studied in 103 erysipelas patients and in 46 persons having had the disease at the acute period of this infection and at the periods between relapses. The elevated levels of CIC and SH-groups of serum deproteinate were found to be directly correlated with the inhibition index. The study showed that, as a rule, in patients with the elevated level of CIC the frequently relapsing form of erysipelas, accompanied by the formation of relative hypersuppressor-type secondary immunodeficiency and by a decrease in the functional activity of dermal macrophages, was observed.


A nonsense mutation in myelin protein zero causes congenital hypomyelination neuropathy through altered P0 membrane targeting and gain of abnormal function.
Protein zero (P0) is the major structural protein in peripheral myelin, and mutations in the Myelin Protein Zero (Mpz) gene produce wide-ranging hereditary neuropathy phenotypes. To gain insight in the mechanisms underlying a particularly severe form, congenital hypomyelination (CH), we targeted mouse Mpz to encode P0Q215X, a nonsense mutation associated with the disease, that we show escapes nonsense mediated decay and is expressed in CH patient nerves. The knock-in mice express low levels of the resulting truncated protein, producing a milder phenotype when compared to patients, allowing to dissect the subtle pathogenic mechanisms occurring in otherwise very compromised peripheral myelin. We find that P0Q215X does not elicit an unfolded protein response, which is a key mechanism for other pathogenic MPZ mutations, but is instead in part aberrantly trafficked to non-myelin plasma membranes and induces defects in radial sorting of axons by Schwann cells. We show that the loss of the C-terminal Tyr-Ala-Met-Leu motif is responsible for P0 mislocalization, as its addition is able to restore correct P0Q215X trafficking in vitro. Lastly, we show that P0Q215X acts through dose-dependent gain of abnormal function, as wild-type P0 is unable to rescue the hypomyelination phenotype. Collectively, these data indicate that alterations at the premyelinating stage, linked to altered targeting of P0, may be responsible for CH, and that different types of gain of abnormal function produce the diverse neuropathy phenotypes associated with MPZ, supporting future allele-specific therapeutic silencing strategies.


The Jak family tyrosine kinase Jak3 is required for IL-2 synthesis by naive/resting CD4+ T cells.
The Jak family tyrosine kinase, Jak3, is involved in signaling through cytokine receptors using the common gamma-chain. Mice deficient in Jak3 have mature T cells, all of which have an activated/memory cell phenotype but are unresponsive to in vitro stimulation. Due to this activated phenotype, it has been impossible to determine whether Jak3 plays a role in the responsiveness of naive/resting T cells. To circumvent this difficulty, we generated naive/resting Jak3-negative T cells by two genetic approaches. After stimulation, these cells failed to produce significant amounts of IL-2. Although no signaling defect could be detected, we did find that naive/resting Jak3-negative T cells have substantially reduced levels of the transcription factor NF-AT1 and moderately reduced levels of c-Jun and c-Fos. On the basis of these data, we propose that Jak3-dependent cytokine signals may be required to maintain the normal levels of basal transcription factors required for immediate responsiveness to Ag activation.


Development of methodology for conducting clinical trials in frontotemporal lobar degeneration.
To design clinical trials for the frontotemporal lobar degenerations (FTLD), knowledge about measurement of disease progression is needed to estimate power and enable the choice of optimal outcome measures. The aim here was to conduct a multicentre, 1 year replica of a clinical trial in patients with one of four FTLD syndromes, behavioural variant frontotemporal dementia (bvFTD), progressive nonfluent aphasia (PNFA), progressive logopenic aphasia (PLA) and semantic dementia (SMD). Patients with one of the four FTLD syndromes were recruited from five academic medical centres over a 2 year period. Standard operationalized diagnostic criteria were used. In addition to clinical inclusion and exclusion criteria, patients were required to exhibit focal frontal, temporal or insular brain atrophy or dysfunction by neuroimaging. Patients underwent neuropsychological, functional, behavioural, neurological and MR imaging assessment at baseline and approximately 12 months later. Potential outcome measures were examined for their rates of floor and ceiling values at baseline and end of study, their mean changes and variances. The neuropsychological tests were combined into two cognitive composites -- one for language functions and the other for executive functions. There were 107 patients who underwent baseline assessment and 78 who completed a follow-up assessment within 10-16 months. Two global measures, the FTLD-modified Clinical Dementia Rating (FTLD-modified CDR) and the Clinical Global Impression of Change (CGIC) demonstrated decline in the majority of patients. Several cognitive measures showed negligible floor or ceiling scores either at baseline or follow-up. Scores declined at follow-up in the majority of patients. The cognitive, executive and combined composites were shown to be sensitive to change across all FTLD syndromes. Patients improved at follow-up on the behavioural scales -- the Frontal Behavioural Inventory (22%) and the Neuropsychiatric Inventory (28%) -- suggesting that these instruments may not be ideal for clinical trial use. It was feasible to recruit FTLD patients in a simulated multi-centre trial. There are several candidate outcome measures -- including the FTLD-CDR and the cognitive composites -- that could be used in clinical trials across the spectrum of FTLD.