The water environment as a source of potentially pathogenic mycobacteria.
Nontuberculous mycobacteria (NTM) are ubiquitous organisms of a wide variety of environmental reservoirs, including natural and municipal water, soil, aerosols, protozoans, animals and humans. Several of these species are potential pathogens which affect human health. The aim of this study was to determine the occurrence of NTM in the water environment. Samples were taken from 13 water-related facilities including fish ponds, storage ponds, drinking water reservoirs and an experimental recirculation system. Altogether, 396 samples of water, sediment and aquatic plants were collected and analysed. All samples were examined using conventional culture methods. Suspected microbial isolates were subjected to polymerase chain reaction analysis and identified using partial sequence analysis of the 16S rDNA gene. The culture revealed 94/396 samples (23.7%) that contained mycobacteria. Among known NTM we identified potentially pathogenic mycobacteria isolated from the fresh water environment for the first time: Mycobacterium asiaticum, M. chimaera, M. interjectum, M. kumamotonense, M. lentiflavum, M. montefiorense, M. nebraskense, M. paraffinicum and M. simiae. Epidemiologic studies suggest that the natural water environment is the principal source of human exposure. Our results indicate that besides the well-known potentially pathogenic mycobacteria it is important to observe occurrence, proliferation and persistence of newly discovered mycobacterial species.


Simulating the approximate irregular field dose distribution in radiotherapy using an ultrasound tracking technique.
This study used an ultrasound image tracking algorithm (UITA) in combination with a proposed simulation program for the approximate irregular field dose distribution (SPAD) to assess the feasibility of performing dose distribution simulations for two-dimensional radiotherapy. This study created five different types of multileaf collimator openings, and applied a SPAD to analyze the matrix position parameters for each regular field to generate a static program-simulation dose distribution map (PDDM), whose similarity was then compared with a static radiochromic film experimental-measurement dose distribution map (EDDM). A two-dimensional respiration motion simulation system (RMSS) was used to reproduce the respiration motion, and the UITA was used to capture the respiration signals. Respiration signals were input to the SPAD to generate two dynamic PDDMs, which were compared for similarity with the dynamic EDDM. In order to verify the dose distribution between different dose measurement techniques, the gamma passing rate with 2%/2 mm criterion was used for the EDDM and PDDM, the passing rates were between 94.31% and 99.71% in the static field analyses, and between 84.45% and 96.09% for simulations with the UITA signal input and between 89.35% and 97.78% for simulations with the original signal input in the dynamic field analyses. Static and dynamic dose distribution maps can be simulated based on the proposed matrix position parameters of various fields and by using the UITA to track respiration signals during radiation therapy. The present findings indicate that it is possible to develop a reusable and time-saving dose distribution measurement tool.


Optimized EPI for fMRI using a slice-dependent template-based gradient compensation method to recover local susceptibility-induced signal loss.
Most functional magnetic resonance imaging (fMRI) experiments use gradient-echo echo planar imaging (GE EPI) to detect the blood oxygenation level-dependent (BOLD) effect. This technique may fail in the presence of anatomy-related susceptibility-induced field gradients in the human head. In this work, we present a novel 3D compensation method in combination with a template-based correction that can be optimized over particular regions of interest to recover susceptibility-induced signal loss without acquisition time penalty. Based on an evaluation of B(0) field maps of eight subjects, slice-dependent gradient compensation moments are derived for maximal BOLD sensitivity in two compromised regions: the orbitofrontal cortex and the amygdala areas. A modified EPI sequence uses these additional gradient moments in all three imaging directions. The method is compared to non-compensated, template-based and subject-specific correction gradients and also in a breath-holding experiment. The slice-dependent gradient compensation method significantly improves signal intensity/BOLD sensitivity by about 35/43% in the orbitofrontal cortex and by 17/30% in the amygdala areas compared to a conventional acquisition. Template-based correction and subject-specific correction perform equally well. The BOLD sensitivity in the breath hold experiment is effectively increased in compensated regions. The new method addresses the problem of susceptibility-induced signal loss, without compromising temporal resolution. It can be used for event-related functional experiments without requiring additional subject-specific calibration or calculation time.


Tuning of resonance-spacing in a traveling-wave resonator device.
In this work a traveling-wave resonator device is proposed and experimentally demonstrated in silicon-on-insulator platform in which the spacing between its adjacent resonance modes can be tuned. This is achieved through the tuning of mutual coupling of two strongly coupled resonators. By incorporating metallic microheaters, tuning of the resonance-spacing in a range of 20% of the free-spectral-range (0.4nm) is experimentally demonstrated with 27mW power dissipation in the microheater. To the best of our knowledge this is the first demonstration of the tuning of resonance-spacing in an integrated traveling-wave-resonator. It is also numerically shown that these modes exhibit high field-enhancements which makes this device extremely useful for nonlinear optics and sensing applications.


Comparison of three instruments to assess changes of motor impairment in acute hemispheric stroke: the Stroke Impairment Assessment Set (SIAS), the National Institute of Health Stroke Scale (NIHSS) and the Canadian Neurological Scale (CNS).
The purpose was to compare the performance of three representative instruments to measure changes of motor recovery with acute hemispheric stroke. In 41 consecutive patients with acute hemispheric infarction, motor recovery was assessed within 3 days of onset and then every 2 weeks until the 12th week with the Stroke Impairment Assessment Set (SIAS), the National Institutes of Health stroke scale (NIHSS) and the Canadian Neurological Scale (CNS). We analyzed the relationships among the corresponding subscales of the three instruments with the Spearman's rank correlation method, and compared their responsiveness by plotting the temporal profiles of scores of each instrument and by testing the significance of changes over time with the Friedman test. High correlations were observed among the three instruments at each examination point. However, the SIAS scores were more widely dispersed at the same NIHSS and CNS scores. Friedman test revealed that the scores changed significantly during the observation period with the SIAS and the CNS but not with the NIHSS. The changes were detected later with the CNS than with the SIAS. Although the assessment results with the three instruments were highly inter-correlated, the SIAS performed better with respect to responsiveness to changes. Implication for Rehabilitation When providing rehabilitation services to patients with stroke, it is important to document objectively the level of their impairment from the acute stage and during the entire course of rehabilitation with standardized instrument, and to predict their functional outcomes as early and accurately as possible. This study therefore compared three representative instruments with established psychometric properties, the SIAS, the NIHSS and the CNS, with respect to their ability to document motor impairment and their responsiveness to recovery in patients with acute hemispheric stroke.


Protein composition of interband regions in polytene and cell line chromosomes of Drosophila melanogaster.
Despite many efforts, little is known about distribution and interactions of chromatin proteins which contribute to the specificity of chromomeric organization of interphase chromosomes. To address this issue, we used publicly available datasets from several recent Drosophila genome-wide mapping and annotation projects, in particular, those from modENCODE project, and compared molecular organization of 13 interband regions which were accurately mapped previously. Here we demonstrate that in interphase chromosomes of Drosophila cell lines, the interband regions are enriched for a specific set of proteins generally characteristic of the "open" chromatin (RNA polymerase II, CHRIZ (CHRO), BEAF-32, BRE1, dMI-2, GAF, NURF301, WDS and TRX). These regions also display reduced nucleosome density, histone H1 depletion and pronounced enrichment for ORC2, a pre-replication complex component. Within the 13 interband regions analyzed, most were around 3-4 kb long, particularly those where many of said protein features were present. We estimate there are about 3500 regions with similar properties in chromosomes of D. melanogaster cell lines, which fits quite well the number of cytologically observed interbands in salivary gland polytene chromosomes. Our observations suggest strikingly similar organization of interband chromatin in polytene chromosomes and in chromosomes from cell lines thereby reflecting the existence of a universal principle of interphase chromosome organization.


Sanitation, Disease Externalities and Anaemia: Evidence From Nepal.
Anaemia impairs physical and cognitive development in children and reduces human capital accumulation. The prior economics literature has focused on the role of inadequate nutrition in causing anaemia. This article is the first to show that sanitation, a public good, significantly contributes to preventing anaemia. We identify effects by exploiting rapid and differential improvement in sanitation across regions of Nepal between 2006 and 2011. Within regions over time, cohorts of children exposed to better community sanitation developed higher haemoglobin levels. Our results highlight a previously undocumented externality of open defaecation, which is today practiced by over a billion people worldwide.


Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-kappa B.
NF-kappa B, which consists of two polypeptides, p50 (M(r) 50K) and p65/RelA (M(r) 65K), is thought to be a key regulator of genes involved in responses to infection, inflammation and stress. Indeed, although developmentally normal, mice deficient in p50 display functional defects in immune responses. Here we describe the generation of mice deficient in the RelA subunit of NF-kappa B. Disruption of the relA locus leads to embryonic lethality at 15-16 days of gestation, concomitant with a massive degeneration of the liver by programmed cell death or apoptosis. Embryonic fibroblasts from RelA-deficient mice are defective in the tumour necrosis factor (TNF)-mediated induction of messenger RNAs for I kappa B alpha and granulocyte/macrophage colony stimulating factor (GM-CSF), although basal levels of these transcripts are unaltered. These results indicate that RelA controls inducible, but not basal, transcription in NF-kappa B-regulated pathways.


Breakthrough attacks in patients with hereditary angioedema receiving long-term prophylaxis are responsive to icatibant: findings from the Icatibant Outcome Survey.
Patients with hereditary angioedema (HAE) due to C1-inhibitor deficiency (C1-INH-HAE) experience recurrent attacks of cutaneous or submucosal edema that may be frequent and severe; prophylactic treatments can be prescribed to prevent attacks. However, despite the use of long-term prophylaxis (LTP), breakthrough attacks are known to occur. We used data from the Icatibant Outcome Survey (IOS) to evaluate the characteristics of breakthrough attacks and the effectiveness of icatibant as a treatment option. Data on LTP use, attacks, and treatments were recorded. Attack characteristics, treatment characteristics, and outcomes (time to treatment, time to resolution, and duration of attack) were compared for attacks that occurred with versus without LTP. Data on 3228 icatibant-treated attacks from 448 patients with C1-INH-HAE were analyzed; 30.1% of attacks occurred while patients were using LTP. Attack rate, attack severity, and the distribution of attack sites were similar across all types of LTP used, and were comparable to the results found in patients who did not receive LTP. Attacks were successfully treated with icatibant; 82.5% of all breakthrough attacks were treated with a single icatibant injection without C1-INH rescue medication. Treatment outcomes were comparable for breakthrough attacks across all LTP types, and for attacks without LTP. Patients who use LTP should be aware that breakthrough attacks can occur, and such attacks can be severe. Thus, patients with C1-INH-HAE using LTP should have emergency treatment readily available. Data from IOS show that icatibant is effective for the treatment of breakthrough attacks. Trial Registration NCT01034969.


Robust scalable high throughput production of monodisperse drops.
Monodisperse drops with diameters between 20 μm and 200 μm can be used to produce particles or capsules for many applications such as for cosmetics, food, and biotechnology. Drops composed of low viscosity fluids can be conveniently made using microfluidic devices. However, the throughput of microfluidic devices is limited and scale-up, achieved by increasing the number of devices run in parallel, can compromise the narrow drop-size distribution. In this paper, we present a microfluidic device, the millipede device, which forms drops through a static instability such that the fluid volume that is pinched off is the same every time a drop forms. As a result, the drops are highly monodisperse because their size is solely determined by the device geometry. This makes the operation of the device very robust. Therefore, the device can be scaled to a large number of nozzles operating simultaneously on the same chip; we demonstrate the operation of more than 500 nozzles on a single chip that produces up to 150 mL h-1 of highly monodisperse drops.


Identification and Quantification of Flavonoids from Two Southern Italian Cultivars of Allium cepa L., Tropea (Red Onion) and Montoro (Copper Onion), and Their Capacity to Protect Human Erythrocytes from Oxidative Stress.
Onions (Allium cepa) are consumed worldwide and represent an important source of dietary phytochemicals with proven antioxidant properties, such as phenolic acids, flavonoids, thiosulfinates, and anthocyanins. Epidemiological and experimental data suggest that regular consumption of onions is associated with a reduced risk of degenerative disorders. Therefore, it is of interest to investigate the biological properties of different varieties of onions. Here, we characterized for the first time a variety of onion, called Ramata di Montoro (coppery onion from Montoro), grown in a niche area in southern Italy, and compared its phenolic profile and antioxidant properties to a commercial ecotype of red onion, Tropea, also present in southern Italy. An analytical method based on high-performance liquid chromatography coupled with UV detection and mass spectrometry was used to separate and characterize the phenolic fraction (anthocyanins and flavonols) extracted from both coppery and red types. The main compounds detected in the two ecotypes were quercetin and quercetin glucosides, isorhamnetin glucosides, kaempferol glucoside, and, among anthocyanins, cyanidin glucosides. Tropea ecotype onion showed a higher content of flavonols (632.82 mg/kg fresh weight) than Montoro type onion (252.91 mg/kg fresh weight). Accordingly, the antioxidant activity of the former was 2.8-fold higher compared to the latter. More pronounced were the differences existing between the four anthocyanins detected in the two ecotypes, with those in the Tropea ecotype onion present at concentrations 20-230-fold higher than in the Montoro type onion. Both extracts reduced LDL oxidation about 6-fold and protected human erythrocytes from oxidative damage induced by HClO by about 40%. In addition, as a consequence of HClO treatment, glutathione concentration in erythrocytes was reduced about 50% and pretreatment with onion extracts induced a recovery of glutathione level by about 15-22%. Qualitative differences highlighted in the chemical composition of the two phenolic extracts, especially the total content of anthocyanins, which was 30-fold higher in Montoro type onion compared to Tropea ecotype, can be associated with the protective effects measured against oxidative damage induced in human erythrocytes.


Metabolic effects associated with atypical antipsychotic treatment in the developmentally disabled.
Atypical antipsychotics, especially clozapine and olanzapine, have been increasingly associated with weight gain and other adverse metabolic events (diabetes mellitus, hyperlipidemia) in non-mentally retarded populations. This report explores the incidence of this phenomenon in an institution-dwelling population of individuals with developmental disabilities. A retrospective longitudinal analysis was performed for a sample of 41 adults with developmental disabilities and comorbid psychiatric and/or behavioral syndromes for whom treatment was converted from typical antipsychotics to olanzapine or risperidone for a minimum period of 2 years. Data were collected from October 1998 to September 2002. Among parameters analyzed were chlorpromazine equivalent dosage of antipsychotic, metabolic parameters, body mass index (BMI), level of concurrent medications, and concomitant dietary restrictions. Thirty-two study subjects (78.0%) were men. The mean age of the study subjects was 43.6 years (at the end of the study). Thirty-seven (90.2%) had severe-to-profound mental retardation. Eight (19.5%) were on a restricted diet. Twenty-three subjects (56.1%) were switched from a typical antipsychotic to olanzapine, and 18 subjects (43.9%) were switched from a typical antipsychotic to risperidone. Of the subsample of subjects who were switched from a typical antipsychotic to risperidone, 12 (66.7%) went on to be switched to olanzapine because of either emergent side effects or lack of efficacy. For the overall sample (N = 41), there was a 19.3% increase in chlorpromazine-equivalent antipsychotic dosage from baseline to the 2-year endpoint along with a 5.6% decrease in fasting blood glucose from baseline to the 2-year endpoint. There were no significant differences between baseline and endpoint values for BMI, total cholesterol, low-density lipoprotein cholesterol, or triglycerides. The findings of this 2-year evaluation suggest that clinically or statistically significant BMI increases as well as blood glucose and lipid elevations are not unavoidably correlated with the use of the atypical antipsychotic agents olanzapine and risperi-done and may be minimized by careful monitoring, a regimen of dietary control, and a moderate activity level in a residential population of individuals with mental retardation.


Nanomaterial based electrochemical sensors for the safety and quality control of food and beverages.
The issue of foodborne related illnesses due to additives and contaminants poses a significant challenge to food processing industries. The efficient, economical and rapid analysis of food additives and contaminants is therefore necessary in order to minimize the risk of public health issues. Electrochemistry offers facile and robust analytical methods, which are desirable for food safety and quality assessment over conventional analytical techniques. The development of a wide array of nanomaterials has paved the way for their applicability in the design of high-performance electrochemical sensing devices for medical diagnostics and environment and food safety. The design of nanomaterial based electrochemical sensors has garnered enormous attention due to their high sensitivity and selectivity, real-time monitoring and ease of use. This review article focuses predominantly on the synthesis and applications of different nanomaterials for the electrochemical determination of some common additives and contaminants, including hydrazine (N2H4), malachite green (MG), bisphenol A (BPA), ascorbic acid (AA), caffeine, caffeic acid (CA), sulfite (SO32-) and nitrite (NO2-), which are widely found in food and beverages. Important aspects, such as the design, fabrication and characterization of graphene-based materials, gold nanoparticles, mono- and bimetallic nanoparticles and metal nanocomposites, sensitivity and selectivity for electrochemical sensor development are addressed. High-performance nanomaterial based electrochemical sensors have and will continue to have myriad prospects in the research and development of advanced analytical devices for the safety and quality control of food and beverages.


Strength and Cardiorespiratory Exercise Rehabilitation for Severely Burned Patients During Intensive Care Units: A Survey of Practice.
Minimizing the deconditioning of burn injury through early rehabilitation programs (RP) in the intensive care unit (ICU) is of importance for improving the recovery time. The aim of this study was to assess current standard of care (SOC) for early ICU exercise programs in major burn centers. We designed a survey investigating exercise RP on the ICU for burn patients with >30% total burned surface area. The survey was composed of 23 questions and submitted electronically via SurveyMonkey® to six major (pediatric and adult) burn centers in Texas and California. All centers responded and reported exercise as part of their RP on the ICU. The characteristics of exercises implemented were not uniform. All centers reported to perform resistive and aerobic exercises but only 83% reported isotonic and isometric exercises. Determination of intensity of exercise varied with 50% of centers using patient tolerance and 17% using vital signs. Frequency of isotonic, isometric, aerobic, and resistive exercise was reported as daily by 80%, 80%, 83%, and 50% of centers, respectively. Duration for all types of exercises was extremely variable. Mobilization was used as a form of exercise by 100% of burn centers. Our results demonstrate that although early RP seem to be integral during burn survivor's ICU stay, no SOC exists. Moreover, early RP are inconsistently administered and large variations exist in frequency, intensity, duration, and type of exercise. Thus, future prospective studies investigating the various components of exercise interventions are needed to establish a SOC and determine how and if early exercise benefits the burn survivor.


Low intake of fruits, berries and vegetables is associated with excess mortality in men: the Kuopio Ischaemic Heart Disease Risk Factor (KIHD) Study.
Diets rich in fruits and vegetables have been of interest because of their potential health benefits against chronic diseases such as cardiovascular disease (CVD) and cancer. The aim of this work was to assess the association of the dietary intake of a food group that includes fruits, berries and vegetables with all-cause, CVD-related and non-CVD-related mortality. The subjects were Finnish men aged 42-60 y examined in 1984-1989 in the prospective Kuopio Ischaemic Heart Disease Risk Factor (KIHD) Study. Dietary intakes were assessed by 4-d food intake record during the baseline phase of the KIHD Study. The risk of all-cause and non-CVD-related deaths was studied in 2641 men and the risk of CVD-related death in 1950 men who had no history of CVD at baseline. During a mean follow-up time of 12.8 y, cardiovascular as well as noncardiovascular and all-cause mortality were lower among men with the highest consumption of fruits, berries and vegetables. After adjustment for the major CVD risk factors, the relative risk for men in the highest fifth of fruit, berry and vegetable intake for all-cause death, CVD-related and non-CVD-related death was 0.66 [95% confidence interval (CI) 0.50-0.88], 0.59 (0.33-1.06), and 0.68 (0.46-1.00), respectively, compared with men in the lowest fifth. These data show that a high fruit, berry and vegetable intake is associated with reduced risk of mortality in middle-aged Finnish men. Consequently, the findings of this work indicate that diets that are rich in plant-derived foods can promote longevity.


Portal hypertension in primary biliary cholangitis: prevalence, natural history and histological correlates.
The histopathological mechanisms underlying portal hypertension in primary biliary cholangitis (PBC) are poorly understood, as is its natural history. We have therefore determined the prevalence, severity and progression of portal hypertension in PBC and investigated whether its presence is related to specific histological lesions. Hepatic venous pressure gradient (HVPG) was measured in 86 patients, with 186 assessments over up to 7 years of follow-up and the results correlated with a semiquantitative grading of 8 histological features and nodular regenerative hyperplasia (NRH). Portal hypertension (HVPG >5 mmHg) was present in 88% of all assessments (86% at baseline), and in 45% of patients at baseline was >12 mmHg (high-risk portal hypertension). The rise in portal pressure occurs early in the disease, since 45% of patients with normal serum bilirubin had a raised HVPG, as did 72% of patients with early (Ludwig stages 1 and 2) disease. After baseline, there was a small increase in HVPG over the next 5 years in most patients. In patients with precirrhotic PBC, 82% had portal hypertension and in 34% this was >12 mmHg. Portal pressure correlated significantly with a semiquantitative grading of cholestasis, interface hepatitis and portal tract and sinusoidal fibrosis. NRH was present in only 20% of wedge biopsies. Portal hypertension commences in the early stages of PBC, long preceding both rises in serum bilirubin and the development of cirrhosis. Around 34% of precirrhotic PBC patients have 'high-risk' portal hypertension, which is associated with lesions in the portal tracts and sinusoids rather than with NRH.


Phase IIa clinical trial of complement C3 inhibitor AMY-101 in adults with periodontal inflammation.
BackgroundGingivitis and periodontitis are prevalent inflammatory diseases of the periodontal tissues. Current treatments are often ineffective or do not prevent disease recurrence. Uncontrolled complement activation and the resulting chronic gingival inflammation are hallmarks of periodontal diseases. We determined the efficacy and safety of a complement 3-targeted therapeutic, AMY-101, which was locally administered to adult patients with periodontal inflammation.MethodsThirty-two patients with gingival inflammation were enrolled in a randomized, placebo-controlled, double-blind, split-mouth phase IIa trial that followed a dose escalation study to select a safe and effective dose in an additional 8 patients. Half of the patient's mouth was randomly assigned to AMY-101 (0.1 mg/site) or placebo injections at sites of inflammation, administered on days 0, 7, and 14, and then evaluated for safety and efficacy outcomes on days 28, 60, and 90. The primary efficacy outcome was a change in gingival inflammation, measured by a modified gingival index (MGI), and secondary outcomes included changes in bleeding on probing (BOP), the amount of plaque, pocket depth, clinical attachment level, and gingival crevicular fluid levels of matrix metalloproteinases (MMPs) over 90 days.ResultsA once-weekly intragingival injection of AMY-101 for 3 weeks was safe and well tolerated in all participants and resulted in significant (P < 0.001) reductions in clinical indices measuring gingival inflammation (MGI and BOP). AMY-101 significantly (P < 0.05) reduced MMP-8 and MMP-9 levels, indicators of inflammatory tissue destruction. These therapeutic effects persisted for at least 3 months after treatment.ConclusionAMY-101 treatment resulted in a significant and sustainable reduction in gingival inflammation without adverse events and, we believe, merits further investigation for the treatment of periodontitis and other oral or peri-implant inflammatory conditions.Trial registrationClinicalTrials.gov identifier NCT03694444.FundingAmyndas Pharmaceuticals.


Effect of socioeconomic inequalities and vulnerabilities on health-system preparedness and response to COVID-19 in Brazil: a comprehensive analysis.
COVID-19 spread rapidly in Brazil despite the country's well established health and social protection systems. Understanding the relationships between health-system preparedness, responses to COVID-19, and the pattern of spread of the epidemic is particularly important in a country marked by wide inequalities in socioeconomic characteristics (eg, housing and employment status) and other health risks (age structure and burden of chronic disease). From several publicly available sources in Brazil, we obtained data on health risk factors for severe COVID-19 (proportion of the population with chronic disease and proportion aged ≥60 years), socioeconomic vulnerability (proportions of the population with housing vulnerability or without formal work), health-system capacity (numbers of intensive care unit beds and physicians), coverage of health and social assistance, deaths from COVID-19, and state-level responses of government in terms of physical distancing policies. We also obtained data on the proportion of the population staying at home, based on locational data, as a measure of physical distancing adherence. We developed a socioeconomic vulnerability index (SVI) based on household characteristics and the Human Development Index. Data were analysed at the state and municipal levels. Descriptive statistics and correlations between state-level indicators were used to characterise the relationship between the availability of health-care resources and socioeconomic characteristics and the spread of the epidemic and the response of governments and populations in terms of new investments, legislation, and physical distancing. We used linear regressions on a municipality-by-month dataset from February to October, 2020, to characterise the dynamics of COVID-19 deaths and response to the epidemic across municipalities. The initial spread of COVID-19 was mostly affected by patterns of socioeconomic vulnerability as measured by the SVI rather than population age structure and prevalence of health risk factors. The states with a high (greater than median) SVI were able to expand hospital capacity, to enact stringent COVID-19-related legislation, and to increase physical distancing adherence in the population, although not sufficiently to prevent higher COVID-19 mortality during the initial phase of the epidemic compared with states with a low SVI. Death rates accelerated until June, 2020, particularly in municipalities with the highest socioeconomic vulnerability. Throughout the following months, however, differences in policy response converged in municipalities with lower and higher SVIs, while physical distancing remained relatively higher and death rates became relatively lower in the municipalities with the highest SVIs compared with those with lower SVIs. In Brazil, existing socioeconomic inequalities, rather than age, health status, and other risk factors for COVID-19, have affected the course of the epidemic, with a disproportionate adverse burden on states and municipalities with high socioeconomic vulnerability. Local government responses and population behaviour in the states and municipalities with higher socioeconomic vulnerability have helped to contain the effects of the epidemic. Targeted policies and actions are needed to protect those with the greatest socioeconomic vulnerability. This experience could be relevant in other low-income and middle-income countries where socioeconomic vulnerability varies greatly. None. For the Portuguese translation of the abstract see Supplementary Materials section.


A reconsideration of long-term-care policy.
This article describes current problems in the financing and delivery of long-term care, and assesses several proposals for reform. Proposed financing changes are found to differ in their objectives, with some reforms seeking to shift from greater to lesser government support, and others seeking to replace current public efforts with expanded entitlements. Organizational reforms hold promise of more equitable allocation of public expenditures for long-term care, but are not likely to reduce such spending. Thus difficult choices cannot be avoided.


Characteristics and health outcomes of patients hospitalised with hypomagnesaemia: a retrospective study from a single centre in the Northern Territory of Australia.
Magnesium is an essential cation in the human body involved in many processes in the human body. Hypomagnesium has been linked to many poor health outcomes. To study the clinical and biochemical characteristics and health outcomes of patients admitted and found to have hypomagnesaemia in an Australian hospital with a high proportion of Indigenous Australians. A retrospective cohort study was conducted of all patients with hypomagnesaemia hospitalised between 1 August 2008 and 31 December 2014 at Royal Darwin Hospital. All relevant demographic, clinical and biochemical data were collected from patients' medical records. The hospital database was reviewed in January 2018 for mortality of all included patients. A total of 876 patients had been admitted with a confirmed diagnosis of hypomagnesaemia during the study period, with mean follow-up period of 4.0 ± 2.7 years. The mean age at admission was 52.4 ± 19.1 years, 52.2% were females and 56.5% were Indigenous Australians. Chronic kidney disease (99.7%), excessive alcohol consumption (45.7%), hypertension (43.9%) and respiratory diseases (15.0%) were the most common conditions in these patients. Hypomagnesaemia was associated with prolonged length of hospital stay. Most patients did not receive treatment for hypomagnesaemia during admission. During the follow-up period, 38.6% of patients died, and the most common causes of death were malignancies (29.9%). Hypomagnesaemia was a common and undertreated condition in hospitalised patients and was associated with poor health outcomes. Therefore, hospitals should develop guidelines for replacing and monitoring magnesium levels during hospitalisation, achieving better outcomes.


Hollow-core photonic crystal fiber-optic probes for Raman spectroscopy.
We have implemented a new Raman fiber-optic probe design based on a hollow-core photonic-crystal excitation fiber surrounded by silica-core collection fibers. The photonic-crystal fiber offers low attenuation at the pump radiation wavelength, mechanical flexibility, high radiation stability, and low background noise. Because the excitation beam is transmitted through air inside the hollow-core fiber, silica Raman scattering is much reduced, improving the quality of the spectra obtained using probes of this design. Preliminary results show that the new probe design decreases the Raman background from the silica by approximately an order of magnitude compared to solid-core silica Raman probes.


Massive transfusion protocols: the role of aggressive resuscitation versus product ratio in mortality reduction.
Exsanguinating hemorrhage necessitating massive blood product transfusion is associated with high mortality rates. Recent data suggest that altering the fresh frozen plasma to packed red blood cell ratio (FFP:PRBC) results in significant mortality reductions. Our purpose was to evaluate mortality and blood product use in the context of a newly initiated massive transfusion protocol (MTP). In July 2005, our American College of Surgeons-verified Level I trauma center implemented an MTP supporting a 1:1.5 FFP:PRBC ratio, improved communications, and enhanced systems flow to optimize rapid blood product availability. During the 4 years surrounding protocol implementation, we reviewed data on trauma patients directly admitted through the emergency department and requiring 10 or more units PRBCs during the first 24 hours. For the 2 years before and subsequent to MTP initiation, there were 4,223 and 4,414 trauma activations, of which 40 and 37 patients, respectively, met study criteria. The FFP:PRBC ratios were identical, at 1:1.8 and 1:1.8 (p = 0.97). Despite no change in FFP:PRBC ratio, mortality decreased from 45% to 19% (p = 0.02). Other significant findings included decreased mean time to first product: cross-matched RBCs (115 to 71 minutes; p = 0.02), FFP (254 to 169 minutes; p = 0.04), and platelets (418 to 241 minutes; p = 0.01). MTP implementation is associated with mortality reductions that have been ascribed principally to increased plasma use and decreased FFP:PRBC ratios. Our study found a significant reduction in mortality despite unchanged FFP:PRBC ratios and equivalent overall mean numbers of transfusions. Our data underscore the importance of expeditious product availability and emphasize that massive transfusion is a complex process in which product ratio and time to transfusion represent only the beginning of understanding.


Immunobiology of Merkel cell carcinoma: implications for immunotherapy of a polyomavirus-associated cancer.
Merkel cell carcinoma (MCC) is an aggressive skin malignancy with a high mortality rate and an increasing incidence. The recent discovery of Merkel cell polyomavirus has revolutionized our understanding of MCC pathogenesis. Viral oncoproteins appear to play a critical role in tumor progression and are expressed in the majority of MCC tumors. Virus-specific humoral and cellular immune responses are detectable in MCC patients and are linked to the natural history of the disease. Despite persistent expression of immunogenic viral proteins, however, MCC tumors are able to evade the immune system. Understanding of the mechanisms of immune evasion employed by MCC tumors is rapidly increasing and offers opportunities for development of rational immune therapies to improve patient outcomes. Here we review recent discoveries in MCC with a special focus on the pathogenic role of Merkel cell polyomavirus and the immunobiology of this virus-associated disease.


Type 2 inflammation and biological therapies in asthma: Targeted medicine taking flight.
The field of asthma has undergone a dramatic change in recent years. Advances in our understanding of type 2 airway inflammation have driven the discovery of monoclonal antibodies targeting specific aspects of the immune pathway. In landmark trials, these drugs have shown efficacy in reducing asthma attacks and exposure to oral corticosteroids, important causes of morbidity in people with asthma. Our review explores the key features of type 2 inflammation in asthma and summarizes the clinical trial evidence of the novel monoclonal antibody treatments and future avenues for treatment.


Sedative Load in Community-Dwelling Older Adults with Mild-Moderate Alzheimer's Disease: Longitudinal Relationships with Adverse Events, Delirium and Falls.
Older adults are frequently prescribed medications with sedative effects, which are associated with numerous adverse consequences. However, the prevalence and longitudinal associations of sedative medication use in community-dwelling older adults with mild-moderate Alzheimer's disease (AD) has not been explored to date. Our objective was to assess the prevalence of sedative medication use in community-dwelling older adults with mild-moderate AD and examine the longitudinal association between sedative medication use and adverse events (AEs). The association between baseline sedative load (SL) and AEs, unscheduled healthcare utilisation, delirium and falls was assessed in older adults with mild-moderate AD over 18 months using secondary analysis of NILVAD trial data (collected from 2014 to 2016). Baseline medication use was assessed, and the SL model was applied to each participant's medication individually. The SL model classifies medications into one of four categories: (1) primary sedatives, (2) medications with a sedating component or prominent side effect, (3) medications with sedation as a potential adverse reaction and (4) all other medications with no known sedative side effects. Medications in group 1 were assigned an SL score of 2, those in group 2 were assigned an SL score of 1, and those in categories 3 and 4 an SL score of 0. SL scores for each medication participants were taking were summed and the total SL calculated as an arithmetic sum of individual medications score. A total SL score ≥ 3 was classed as high. Statistical analysis was conducted using Poisson regression and mixed-effects linear regression, with adjustment for important clinical covariates. We also assessed the impact of SL on dementia progression and cognitive decline. Over half (55.7% [284/510]) of those with mild-moderate AD (age 72.8 ± 8.3 years, 61.9% female) were prescribed a regular medication with sedation as a primary effect or prominent side effect, with 22.2% (113/510) having a high SL (≥ 3). The most common medications contributing to SL were antidepressants, antipsychotics, anxiolytics and hypnotics. Over 18 months, increasing baseline SL was associated with incident AEs (incidence rate ratio [IRR] 1.15; 95% confidence interval [CI] 1.11-1.19; p < 0.001), serious AEs (IRR 1.23; 95% CI 1.11-1.36; p < 0.001) and unscheduled general practitioner visits (IRR 1.23; 95% CI 1.13-1.34; p < 0.001). Further, increasing SL was associated with a greater likelihood of incident delirium (IRR 1.30; 95% CI 1.11-1.53; p < 0.001) and falls (IRR 1.20; 95% CI 1.03-1.42; p = 0.02). Associations persisted after robust covariate adjustment. SL was not associated with accelerated cognitive decline or AD progression. In the current study, over half of older adults with mild-moderate AD were prescribed at least one drug with a sedative effect, and a significant minority had a high SL. Increasing baseline SL was associated with a greater likelihood of incident AEs, delirium and falls, highlighting the need for optimal prescribing in this potentially vulnerable cohort.


Impact of Surgical Waiting-List Times on Scoliosis Surgery: The Surgeon's Perspective.
Survey. The aim of this study was to evaluate the surgeon's perspective on the potential impact of prolonged surgical waitlists on the surgical care and perioperative management of patients with scoliosis. The long waits for surgical treatment of scoliosis found in some countries may have serious implications for the complexity of surgery and perioperative care required if the curve progresses while waiting. The surgeon's perspective on this problem provides important information that needs to be taken into account during resource allocation. Radiographs from 13 patients who had waited more than 6 months for scoliosis surgery were selected. Each patient had radiographs from the time of surgical booking and immediately preoperatively. The radiographs and a questionnaire were sent to 3 surgeons to canvass their surgical and postoperative plan. The surgeons were blinded to the fact that the radiographs were of the same patients at 2 time points. The patients' actual course of treatment was documented. Data for 11 patients were available for analysis. The average wait for surgery was 24 months (range, 17-30 mo). The mean curve progression was 25.3° while on the waitlist, from an average of 52° to 77°. By the time the patients had to undergo surgery, more anterior releases were added to posterior instrumentation alone in the surgical plan. Mean estimated operative time increased by 2.2 hours, mean estimated length of hospital stay increased by 1 day, and the estimated level of difficulty of surgery increased 2.33 grades. The predicted estimated blood loss also increased. From the surgeon's perspective, lengthy waitlists have a significant negative impact on the perioperative and postoperative care of patients with scoliosis by increasing the complexity of surgery. The actual course of treatment corresponded to the responses from these different surgeons. N/A.


Variation in nicotinic acetylcholine receptor genes is associated with multiple substance dependence phenotypes.
There is shared genetic risk for dependence on multiple substances, and the nicotinic receptor gene cluster on chromosome 15 harbors multiple polymorphisms that associate to this risk. Here, we report the results of an association study with 21 SNPs genotyped across the CHRNA5, CHRNA3, and CHRNB4 loci on chromosome 15q25.1. The sample consists of a discovery set (N=1858) of European-American and African-American (AA) families, ascertained on the basis of a sibling pair with cocaine and/or opioid dependence, and a case-control replication sample (N=3388) collected for association studies of alcohol, cocaine, and opioid dependence. We tested the SNPs for association with lifetime cocaine, opioid, nicotine, and alcohol dependence. We replicated several previous findings, including associations between rs16969968 and nicotine dependence (P=0.002) and cocaine dependence (P=0.02), with opposite risk alleles for each substance. We observed these associations in AAs, which is a novel finding. The strongest association signal in either sample was between rs684513 in CHRNA5 and cocaine dependence (OR=1.43, P=0.0004) in the AA replication set. We also observed two SNPs associated with alcohol dependence, that is, rs615470 in CHRNA5 (OR=0.77, P=0.0006) and rs578776 (OR=0.78, P=0.001). The associations between CD and rs684513, AD and rs615470, and AD and rs578776 remained significant after a permutation-based correction for multiple testing. These data reinforce the importance of variation in the chromosome 15 nicotinic receptor subunit gene cluster for risk of dependence on multiple substances, although the direction of the effects may vary across substances.


Clinical and anatomy study of the human mental foramen.
The great diffusion of the surgical techniques in jaws surgery and the progress of the radiological imagining procedures expressed many interest in clinical anatomy of the mental foramen (MF). The study goal was to determine the precise location of the MF and the surrounding anatomical landmarks. Measurements of the MF position relative to the surgical landmarks and related posterior teeth were made on 20 dry mandibles with complete dentition and intact alveolar bridge obtained from the Institute of Anatomy, School of Medicine, University in Sarajevo. The measurements were made by anthropometric methods on the booth sides of the mandible, and compared with measurement made on the orthopantomogram radiographs of the same mandibles. The most common position of the MF was in line with the longitudinal axis of the second premolar. In the vertical plane on the skulls the MF lays in the midpoint of the distance between the lower border of the mandible and the alveolar margin, however on the orthopantomogram MF appeared slightly bellow the midpoint. In the horizontal plane it lays approximately one third of the distance between the mandibular symphysis and the posterior border of the ramus of the mandible measurement from A-P projection and one quarter of that distance measurement from the profile projection. There were no significant differences between distances MF from posterior border of the ramus of the mandible measurement from A-P and profile projection and the one obtained on orthopantomogram and their ratio is constant value determine das 1,065. The MF was on average 25 mm lateral to the mandibular skeletal midline and symmetrical, and symmetry was preserved on the orthopantomogram. The measurement showed significant differences in distances of M from superior border mandible measurement on dry mandible and orthopantomogram radiographs while distance bellow the MF was not significantly different. The constant values of MF distance to the posterior border of the ramus of the mandible measured as 1,065 and the distance to the medial skeletal line of 2,11 made it possible to also determine average angle of 43 degrees stream of the cor-pus of the mandible behind MF. These values in combination with ratios of MF to the different anatomic landmarks designated as relative horizontal and relative vertical position, would be of importance not only from anatomical but also from practical point of view for estimation of alveolar bridge resorption and preoperative analysis in orthognat postresection or implant surgery in the mandible.


Predictive Score for Posttransplantation Outcomes.
Most current scoring tools to predict allograft and patient survival upon kidney transplantion are based on variables collected posttransplantation. We developed a novel score to predict posttransplant outcomes using pretransplant information including routine laboratory data available before or at the time of transplantation. Linking the 5-year patient data of a large dialysis organization to the Scientific Registry of Transplant Recipients, we identified 15 125 hemodialysis patients who underwent first deceased transplantion. Prediction models were developed using Cox models for (a) mortality, (b) allograft loss (death censored), and (c) combined death or transplant failure. The cohort was randomly divided into a two thirds set (Nd = 10 083) for model development and a one third set (Nv = 5042) for validation. Model predictive discrimination was assessed using the index of concordance, or C statistic, which accounts for censoring in time-to-event models (a-c). We used the bootstrap method to assess model overfitting and calibration using the development dataset. Patients were 50 ± 13 years of age and included 39% women, 15% African Americans, and 36% persons with diabetes. For prediction of posttransplant mortality and graft loss, 10 predictors were used (recipients' age, cause and length of end-stage renal disease, hemoglobin, albumin, selected comorbidities, race and type of insurance as well as donor age, diabetes status, extended criterion donor kidney, and number of HLA mismatches). The new model (www.TransplantScore.com) showed the overall best discrimination (C-statistics, 0.70; 95% confidence interval [95% CI], 0.67-0.73 for mortality; 0.63; 95% CI, 0.60-0.66 for graft failure; 0.63; 95% CI, 0.61-0.66 for combined outcome). The new prediction tool, using data available before the time of transplantation, predicts relevant clinical outcomes and may perform better to predict patients' graft survival than currently used tools.


Myeloid Cell Crosstalk Regulates the Efficacy of the DNA/ALVAC/gp120 HIV Vaccine Candidate.
Vaccination with DNA-SIV + ALVAC-SIV + gp120 alum results in inflammasome activation, high levels of IL-1β production, emergency myelopoiesis, and the egress of CXCR4+ CD14+ pre-monocytes from bone marrow. Previously we have shown that this vaccine-induced innate monocyte memory is associated with decreased risk of SIVmac251 acquisition. Because IL-1β also promotes the propagation of monocyte-derived suppressor (M-MDSC)-like cells, here we extended our analysis to this negative regulator subset, characterizing its levels and functions in macaques. Interestingly, we found that DNA prime engages M-MDSC-like cells and their levels are positively associated with the frequency of CD14+ classical monocytes, and negatively with the levels of CD16+ monocytes, correlates of decreased and increased risk of SIV acquisition, respectively. Accordingly, M-MDSC frequency, arginase activity, and NO were all associated with decrease of CD8 T cells responses and worse vaccination outcome. DNA vaccination thus induces innate immunity by engaging three subsets of myeloid cells, M-MDSCs, CD14+ innate monocyte memory, and CD16+ monocytes all playing different role in protection. The full characterization of the immunological space created by myeloid cell crosstalk will likely provide clues to improve the efficacy of HIV vaccine candidates.


Haptoglobin preserves the CD163 hemoglobin scavenger pathway by shielding hemoglobin from peroxidative modification.
Detoxification and clearance of extracellular hemoglobin (Hb) have been attributed to its removal by the CD163 scavenger receptor pathway. However, even low-level hydrogen peroxide (H(2)O(2)) exposure irreversibly modifies Hb and severely impairs Hb endocytosis by CD163. We show here that when Hb is bound to the high-affinity Hb scavenger protein haptoglobin (Hp), the complex protects Hb from structural modification by preventing alpha-globin cross-links and oxidations of amino acids in critical regions of the beta-globin chain (eg, Trp15, Cys93, and Cys112). As a result of this structural stabilization, H(2)O(2)-exposed Hb-Hp binds to CD163 with the same affinity as nonoxidized complex. Endocytosis and lysosomal translocation of oxidized Hb-Hp by CD163-expressing cells were found to be as efficient as with nonoxidized complex. Hp complex formation did not alter Hb's ability to consume added H(2)O(2) by redox cycling, suggesting that within the complex the oxidative radical burden is shifted to Hp. We provide structural and functional evidence that Hp protects Hb when oxidatively challenged with H(2)O(2) preserving CD163-mediated Hb clearance under oxidative stress conditions. In addition, our data provide in vivo evidence that unbound Hb is oxidatively modified within extravascular compartments consistent with our in vitro findings.


Late dropout from nursing education: An interview study of nursing students' experiences and reasons.
The global shortages of nurses require a closer look at why nursing students stop in the later years of their degree programme. The purpose of this study is to explore nursing students' experiences and reasons that lead to this late dropout. Semi-structured interviews were held in 2017 with eleven former nursing students who dropped out in the third year of their Bachelor's Nursing degree programme in the Netherlands. Data was collected and analysed iteratively, following the principles of Thematic Analysis. Two core themes were identified: 'ending up in a downward spiral of physical, psychological and social problems' and 'experiencing an increasing mismatch between expectations and reality'. Reasons for late dropout from nursing education are diverse and interlinked. In contrast with studies on early dropout, academic difficulties did not play a major role in late dropout. Negative experiences during clinical placements led to dropout in both groups. One group lacked a safe learning environment in clinical placements, study coaching and psychological support. The other group missed realistic information provision about nursing education and the broad range of career opportunities in nursing.


Colorectal and anal neoplasms following liver transplantation.
Liver transplantation (LT) is the treatment of choice for end-stage liver disease. The required immunosuppression increases the risk for developing malignancies. Some viruses play a crucial role. Data on neoplasms of the colon, rectum and anus in LT are limited. A retrospective evaluation of the incidence and clinical course of colorectal and anal malignancies and colonic polyps in a series of 467 consecutive LTs in 402 individuals between 1998 and 2001 was performed. Standard immunosuppression included Tacrolimus, Mycophenolic acid and steroids. During a median follow up of 5.2 years, three colon adenocarcinomas, one EBV associated cecal posttransplant lymphoproliferative tumour and two HPV associated anal tumours were identified. Pre-LT colonoscopy was performed in 161 patients (40%), and of 153 evaluable individuals, 53 (34.9%) had polyps. Colonoscopy was performed in 186 patients (46.3%) median 14.8 (range 0.2-77.8) months post-LT and 55 (29.3%) had polyps. Post-LT adenomatous polyps were detected in 47.3% of patients with pre-LT polyps vs 6.7% of patients without pre-LT polyps (P < 0.001). Patients with alcoholic liver disease had a significantly higher rate of adenoma formation (50.0% vs 11.1%, P < 0.001). No patient died from colorectal/anal malignancy. The incidence of metachronous and new polyp formation in our study is similar to people who are not immunocompromised, but subgroups are at increased risk. Viral-associated malignancies, including post-transplant lymphoproliferative disorders and anal cancer, are important entities in the LT population suggesting that complete screening of the colon, rectum and anus including pre-LT and post-LT colonoscopy should be utilized.


Management of Hypertension in Diabetic Nephropathy: How Low Should We Go?
Hypertension is a frequent comorbidity often following the development of diabetic nephropathy among individuals with type 1 diabetes and affecting most patients with type 2 diabetes at the time of diagnosis. Multiple prospective randomized placebo-controlled trials demonstrate that tight blood pressure control among patients with diabetic nephropathy reduces the rates of macrovascular and microvascular complications. While randomized trials exist and support a blood pressure goal of <140/90 mm Hg for patients with nondiabetic kidney disease, there are no prospective data regarding a specific blood pressure goal on progression of diabetic nephropathy. Retrospective data analyses from trials show a linear relationship between either baseline or achieved study blood pressure and progression of nephropathy. Very high albuminuria is a hallmark of diabetic nephropathy with reductions by either angiotensin converting enzyme inhibitors (ACEi) or angiotensin receptor blocker (ARB) monotherapy associated with slowed nephropathy progression. However, combination antihypertensive therapy, while decreasing proteinuria, augments the risk of hyperkalemia, hypotension, and kidney dysfunction. Given the lack of trial data for a BP goal among patients with diabetic nephropathy, prospective trials are needed to define the optimal blood pressure necessary to preserve kidney function. At present, guideline blood pressure goals of less than 140/90 mm Hg and the use of ACEi or ARB therapy for those with more than 300 mg of albuminuria are mandated.


Schmallenberg virus--is it present in South Africa?
In July 2006, a case of two out of three lambs born to one ewe in a flock of 45 had signs that, in retrospect, were possibly consistent with Schmallenberg virus infection. This occurred in the Onderstepoort area (Gauteng Province) but a definitive diagnosis was not made. Then, in May 2008, a farmer in the Delmas area (Mpumalanga Province) reported that deformed lambs had been born to several ewes in the flock. Six of the approximately 50 mated ewes gave birth to lambs showing varying degrees of arthrogryposis, torticollis, kyphosis, mandibular brachygnathia and hydrocephalus. Of these, only two were born alive but they died within a few hours. Blood was collected from the ewes with deformed lambs, a random sample of ewes that had given birth to normal lambs and a lamb that was normal but had a twin that was deformed. The samples were tested for Wesselsbron and Akabane antibodies using a complement fixation test and a haemagglutination/haemagglutination inhibition test that were available at that time. Bluetongue virus antibodies were also tested for using a commercial Enzyme-linked immunosorbent assay (ELISA) test. All samples showed negative results for all diseases tested. At the time Rift Valley fever virus had not been diagnosed in that region for many years and so it was not included in the testing. It is unlikely that this was the cause as no liver pathology was detected on postmortem examination of the lambs and no adult ewes had died. The farmer reported that another farm just a few kilometres away experienced the same deformities in some of their lambs but this farm was not investigated. During investigation it was thought that the cause was possibly a new strain of Akabane virus, although there was no way to confirm it. However, with the recent discovery of the Schmallenberg virus, it is possible that this virus has been present in South Africa for at least the last four years without being identified.


Intracellular mGluR5 can mediate synaptic plasticity in the hippocampus.
Metabotropic glutamate receptor 5 (mGluR5) is widely expressed throughout the CNS and participates in regulating neuronal function and synaptic transmission. Recent work in the striatum led to the groundbreaking discovery that intracellular mGluR5 activation drives unique signaling pathways, including upregulation of ERK1/2, Elk-1 (Jong et al., 2009) and Arc (Kumar et al., 2012). To determine whether mGluR5 signals from intracellular membranes of other cell types, such as excitatory pyramidal neurons in the hippocampus, we used dissociated rat CA1 hippocampal cultures and slice preparations to localize and characterize endogenous receptors. As in the striatum, CA1 neurons exhibited an abundance of mGluR5 both on the cell surface and intracellular membranes, including the endoplasmic reticulum and the nucleus where it colocalized with the sodium-dependent excitatory amino acid transporter, EAAT3. Inhibition of EAAT3 or sodium-free buffer conditions prevented accumulations of radiolabeled agonist. Using a pharmacological approach to isolate different pools of mGluR5, both intracellular and cell surface receptors induced oscillatory Ca(2+) responses in dissociated CA1 neurons; however, only intracellular mGluR5 activation triggered sustained high amplitude Ca(2+) rises in dendrites. Consistent with the notion that mGluR5 can signal from intracellular membranes, uncaging glutamate on a CA1 dendrite led to a local Ca(2+) rise, even in the presence of ionotropic and cell surface metabotropic receptor inhibitors. Finally, activation of intracellular mGluR5 alone mediated both electrically induced and chemically induced long-term depression, but not long-term potentiation, in acute hippocampal slices. These data suggest a physiologically relevant and important role for intracellular mGluR5 in hippocampal synaptic plasticity.


Evaluation of the impact of RNA preservation methods of spiders for de novo transcriptome assembly.
With advances in high-throughput sequencing technologies, de novo transcriptome sequencing and assembly has become a cost-effective method to obtain comprehensive genetic information of a species of interest, especially in nonmodel species with large genomes such as spiders. However, high-quality RNA is essential for successful sequencing, and sample preservation conditions require careful consideration for the effective storage of field-collected samples. To this end, we report a streamlined feasibility study of various storage conditions and their effects on de novo transcriptome assembly results. The storage parameters considered include temperatures ranging from room temperature to -80°C; preservatives, including ethanol, RNAlater, TRIzol and RNAlater-ICE; and sample submersion states. As a result, intact RNA was extracted and assembly was successful when samples were preserved at low temperatures regardless of the type of preservative used. The assemblies as well as the gene expression profiles were shown to be robust to RNA degradation, when 30 million 150-bp paired-end reads are obtained. The parameters for sample storage, RNA extraction, library preparation, sequencing and in silico assembly considered in this work provide a guideline for the study of field-collected samples of spiders.


Variability of serum markers of erythropoiesis during 6 days of racing in highly trained cyclists.
The athlete biological passport for the fight against doping is currently based on longitudinal monitoring for abnormal changes in cellular blood parameters. Serum parameters related to altered erythropoiesis could be considered for inclusion in the passport. The aim of this study was to quantify the changes in such parameters in athletes during a period of intense exercise.12 highly trained cyclists tapered for 3 days before 6 days of simulated intense stage racing. Morning and afternoon blood samples were taken on most days and analysed for total protein, albumin, soluble transferrin receptor and ferritin concentrations. Plasma volume was determined via total haemoglobin mass measured by carbon-monoxide rebreathing. Percent changes in means from baseline and percent standard errors of measurement (analytical error plus intra-athlete variation) on each measurement occasion were estimated with mixed linear modelling of log-transformed measures. Means of all variables changed substantially in the days following the onset of racing, ranging from -13% (haemoglobin concentration) to +27% (ferritin). After the second day, errors of measurement were generally twice those at baseline. Plasma variables were affected by heavy exercise, either because of changes in plasma volume (total protein, albumin, haemoglobin), acute phase/inflammatory reactions (ferritin) or both (soluble transferrin receptor). These effects need to be taken into consideration when integrating a plasma parameter into the biological passport model for athletes.


Mitochondrial fragmentation affects neither the sensitivity to TNFα-induced apoptosis of Brucella-infected cells nor the intracellular replication of the bacteria.
Mitochondria are complex organelles that participate in many cellular functions, ranging from ATP production to immune responses against viruses and bacteria. This integration of a plethora of functions within a single organelle makes mitochondria a very attractive target to manipulate for intracellular pathogens. We characterised the crosstalk that exists between Brucella abortus, the causative agent of brucellosis, and the mitochondria of infected cells. Brucella replicates in a compartment derived from the endoplasmic reticulum (ER) and modulates ER functionality by activating the unfolded protein response. However, the impact of Brucella on the mitochondrial population of infected cells still requires a systematic study. We observed physical contacts between Brucella containing vacuoles and mitochondria. We also found that B. abortus replication is independent of mitochondrial oxidative phosphorylation and that mitochondrial reactive oxygen species do not participate to the control of B. abortus infection in vitro. We demonstrated that B. abortus and B. melitensis induce a drastic mitochondrial fragmentation at 48 hours post-infection in different cell types, including myeloid and non-myeloid cells. This fragmentation is DRP1-independent and might be caused by a deficit of mitochondrial fusion. However, mitochondrial fragmentation does not change neither Brucella replication efficiency, nor the susceptibility of infected cells to TNFα-induced apoptosis.


Limitations of using population-based databases to assess trends in spinal stereotactic radiosurgery.
There exist no population-based examinations of stereotactic radiosurgery (SRS) of the spine. To address this, spinal SRS was analyzed using the New Jersey State Ambulatory Surgery and Services Database (SASD) and Nationwide Inpatient Sample (NIS) over 10-year periods. The SASD is a state-specific ambulatory surgery and outpatient database, while the NIS database comprises approximately 20% of all nonfederal hospital inpatient admissions and discharges in the United States. Only patients receiving SRS for at least one spinal lesion were included (ICD-9-CM=92.3x; diagnosis codes=63620,63621). 4,194,207 patients were contained in the New Jersey SASD from 2003-2012, of whom fewer than 0.0003% received SRS of any sort, with none receiving SRS of the spine. Of the 78,686,628 patients contained in the NIS, only 16 (0.00002%) received radiosurgery, none of whom received SRS of the spine. In conclusion, a decade-long analysis of the NIS and SASD from the most densely populated state in the United States revealed that no patients received spinal SRS with virtually none receiving SRS of any sort. Given the improbability of these findings, it is much more likely that neither the NIS nor SASD can accurately capture patients receiving SRS. Accurate characterization of the incidence and usage of spinal SRS will require databases less reliant on ICD-9 coding than the SASD or NIS.


Reducing antipsychotic drug use in nursing homes. A controlled trial of provider education.
In the United States, 20% or more of nursing home residents receive antipsychotic drugs, primarily for the behavioral manifestations of dementia. This high level of use of drugs with substantial toxicity has engendered a strong and persistent controversy and recently has led to explicit regulatory measures to curtail use (Omnibus Budget Reconciliation Act of 1987). We developed and tested a comprehensive program to reduce antipsychotic use through education of physicians, nurses, and other nursing home staff. The primary elements of the program were instruction in use of behavioral techniques to manage behavior problems and encouragement of a trial of gradual antipsychotic withdrawal. In a nonrandomized controlled trial, the program was implemented (beginning in August 1990) in two rural Tennessee community nursing homes with elevated antipsychotic use; two other comparable homes were selected as concurrent controls. Throughout the study 194 residents were in the education homes and 184 were in the control homes. Residents in both groups of homes had comparable demographic characteristics and functional status, and each group had a baseline rate of 29 days of antipsychotic use per 100 days of nursing home residence. The primary end points were postintervention changes in administration of antipsychotics and other psychotropic drugs, use of physical restraints, and frequency of behavior problems. Days of antipsychotic use decreased by 72% in the education homes vs 13% in the control homes (P < .001). No significant changes were noted in the use of other psychotropic drugs in either group. Days of physical restraint use decreased 36% in the education homes vs 5% in the control homes (P < .001). Behavior problem frequency did not increase in either group, even among the 48% of baseline antipsychotic users in the education homes who had antipsychotic drug regimens discontinued for 3 or more months. The educational program led to a substantial reduction in antipsychotic use with no increase in the frequency of behavior problems. This suggests that for many antipsychotic drug users benefits may be marginal and that programs to reduce such drug use among the 250,000 US nursing home residents receiving these drugs should have high priority.


Hepatocyte growth factor can substitute for M-CSF to support osteoclastogenesis.
Osteopetrotic mice lacking functional macrophage-colony stimulating factor (M-CSF) recover with ageing, suggesting that alternative osteoclastogenesis pathways exist. Hepatocyte growth factor (HGF) and M-CSF signal through tyrosine kinase receptors and phosphorylate common transducers and effectors such as Src, Grb2, and PI3-Kinase. HGF is known to play a role in osteoclast formation, and in this study we have determined whether HGF could replace M-CSF to support human osteoclastogenesis. We found that the HGF receptor, c-Met, is expressed by the CD14(+) monocyte fraction of human peripheral blood mononuclear cells (PBMC). HGF was able to support monocyte-osteoclast differentiation in the presence of receptor activator for nuclear factor kappaB ligand as evidenced by the formation of numerous multinucleated tartrate-resistant acid phosphatase and vitronectin receptor positive cells which formed F-actin rings and were capable of lacunar resorption. The addition of a neutralising antibody to M-CSF did not inhibit osteoclast differentiation. HGF is a well-established survival factor and viability assays and live/dead staining showed that it promoted the survival and proliferation of monocytes and osteoclasts in a manner similar to M-CSF. Our findings indicate that HGF can substitute for M-CSF to support human osteoclast formation.


The effects of metabolic syndrome versus infectious burden on inflammation, severity of coronary atherosclerosis, and major adverse cardiovascular events.
The clinical predictors of inflammation in atherosclerosis remain controversial. The objective of this study was to compare the associations of metabolic factors vs. infectious burden (IB) with inflammation, the severity of coronary atherosclerosis, and major adverse cardiovascular events (MACEs). Coronary angiography with Gensini score was applied to assess the severity of coronary atherosclerosis in 568 patients with coronary artery disease. Metabolic syndrome (MS) score (0-5) was defined according to the modified criteria of National Cholesterol Education Program Adult Treatment Panel III. IB score (0-7) was defined as the number of seropositivities to several agents. IB score was not associated with plasma C-reactive protein (CRP) concentration, Gensini score, or the risk of MACE. In contrast, MS score significantly correlated with both plasma CRP concentration and Gensini score (P < 0.001 for both). MS score and plasma CRP concentration were also significantly associated with the risk of MACE (hazard ratios 1.51, P < 0.001; and 1.90, P = 0.002, respectively). Compared with IB, metabolic abnormalities have a more prominent association with the degree of inflammation, the severity of coronary atherosclerosis, and the risk of MACE in patients with coronary artery disease.


Different biochemical correlates for different neuropsychiatric abnormalities in patients with cirrhosis.
The diagnosis of hepatic encephalopathy (HE) relies on clinical, neurophysiological, psychometric and laboratory variables. The relationships between such tests remain debated. The aim of this study was to determine the laboratory correlates/prognostic value of neurophysiological/psychometric abnormalities in patients with cirrhosis. Seventy-two patients and 14 healthy volunteers underwent EEG and paper-and-pencil psychometry (PHES). Blood was obtained for C reactive protein (CRP), interleukin 6 (IL6), tumor necrosis factor (TNF)α, ammonia and indole/oxindole. Patients were followed prospectively for a median of 22 months in relation to the occurrence of death, transplantation and HE-related hospitalizations. Thirty-three patients had normal PHES and EEG, 6 had abnormal PHES, 18 abnormal EEG and 13 abnormal PHES and EEG. Patients with abnormal PHES had higher CRP (17 ± 22 vs 7 ± 6, P < 0.01), IL6 (32 ± 54 vs 12 ± 13, P < 0.05) and TNFα (17 ± 8 vs 11 ± 7, P < 0.001) levels than those with normal PHES. Patients with abnormal EEG had higher indole (430 ± 270 vs 258 ± 255, P < 0.01) and ammonia (66 ± 35 vs 45 ± 27, P < 0.05) levels than those with normal EEG. Psychometric test scores showed significant correlations with CRP, TNFα and IL6; EEG indices with ammonia and IL6. CRP and TNFα concentrations were independent predictors of abnormal PHES, ammonia and indole of abnormal EEG on multivariate analysis. Seven patients were lost to follow-up; of the remaining 65, 20 died and 14 underwent transplantation; 15 developed HE requiring hospitalization. PHES and EEG performance were independent predictors of HE and death (P < 0.05). PHES and EEG abnormalities in patients with cirrhosis have partially different biochemical correlates and independently predict outcome.


Precise determination of the dielectric constant and thickness of a nanolayer by use of surface plasmon resonance sensing and multiexperiment linear data analysis.
Surface plasmon resonance (SPR) sensing and an enhanced data analysis technique are used to obtain precise predictions of the dielectric constant and thickness of a nanolayer. In the proposed approach, a modified analytical method is used to obtain initial estimates of the dielectric constants and thicknesses of the metal film and a nanolayer on the sensing surface of a SPR sensor. A multiexperiment data analysis approach based on a two-solvent SPR method is then employed to improve the initial estimates by suppressing the noise in the measurement data. The proposed two-stage approach is employed to determine the dielectric constant and thickness of a molecular imprinting polymer nanolayer. It is found that the results are in good agreement with those obtained with an ellipsometer and a high-resolution scanning electron microscope.


Determination of the index inhomogeneity of transparent isotropic optical material with a dual Sagnac interferometer.
A technique for the measurement of inhomogeneity of optical glass, fused silica, etc., using a Sagnac interferometer (SI) has been presented. An SI produces a pair of laterally separated, mutually parallel, collimated beams with orthogonal planes of polarization, i.e., p and s polarizations from an expanded, linearly polarized (45°) collimated He-Ne (632.8 nm) input laser beam. The p and s beams pass through a liquid-filled cell with plane parallel glass windows. The test glass with plane parallel end surfaces is kept in the path of the p beam in the index matching liquid, while the s beam traverses a parallel path through the liquid. Another SI recombines the emergent p and s beams by removing the lateral shear. A quarter-wave plate transforms the state of polarization of the beams to opposite circular polarizations of which the components selected by a linear polarizer interfere to form Fizeau fringes. Polarization phase shifting interferometery has been applied to determine the optical path difference (OPD) variations. The OPD variation without the test glass is subtracted from that with test glass to eliminate the effect of system aberration. The results for a phosphate laser glass sample have been presented.


Interactions between HIF-1α and AMPK in the regulation of cellular hypoxia adaptation in chronic kidney disease.
Renal hypoxia contributes to chronic kidney disease (CKD) progression, as validated in experimental and human CKD. In the early stages, increased oxygen consumption causes oxygen demand/supply mismatch, leading to hypoxia. Hence, early targeting of the determinants and regulators of oxygen consumption in CKD may alter the disease course before permanent damage ensues. Here, we focus on hypoxia inducible factor-1α (HIF-1α) and AMP-activated protein kinase (AMPK) and on the mechanisms by which they may facilitate cellular hypoxia adaptation. We found that HIF-1α activation in the subtotal nephrectomy (STN) model of CKD limits protein synthesis, inhibits apoptosis, and activates autophagy, presumably for improved cell survival. AMPK activation was diminished in the STN kidney and was remarkably restored by HIF-1α activation, demonstrating a novel role for HIF-1α in the regulation of AMPK activity. We also investigated the independent and combined effects of HIF-1α and AMPK on cell survival and death pathways by utilizing pharmacological and knockdown approaches in cell culture models. We found that the effect of HIF-1α activation on autophagy is independent of AMPK, but on apoptosis it is partially AMPK dependent. The effects of HIF-1α and AMPK activation on inhibiting protein synthesis via the mTOR pathway appear to be additive. These various effects were also observed under hypoxic conditions. In conclusion, HIF-1α and AMPK appear to be linked at a molecular level and may act as components of a concerted cellular response to hypoxic stress in the pathophysiology of CKD.


Kinetics of accumulation, efflux and the pharmacological effects of tritiated dihydroazapetine on the rabbit aorta.
Azepatine, a potent alpha adrenergic antagonist, was catalytically reduced with tritium and hydrogen gas to form dihydroazapetine. The pA2 azapetine was 7.9 whereas that of dihydroazapetine was 6.6, corresponding to a 20-fold decrease in potency. 3H-dihydroazapetine is accumulated into three kinetically distinct compartment in the denervated rabbit aorta. Likewise, efflux of the labeled antagonist occurs from three compartments. The rat constant for onset of alpha adrenoreceptor blockade is 3.15 min-1 which is nearly identical to the rate constant for entry of 3H-dihydroazapetine into a rapidly filling compartment (3.86 min-1) possibly representing the extracellular space. These data are consistent with the hypothesis that onset of alpha adrenoreceptor blockade by dihydroazapetine is diffusion limited.


Acute-on-chronic liver failure: an update.
Acute-on-chronic liver failure (ACLF) is a syndrome characterised by acute decompensation of chronic liver disease associated with organ failures and high short-term mortality. Alcohol and chronic viral hepatitis are the most common underlying liver diseases. Up to 40%-50% of the cases of ACLF have no identifiable trigger; in the remaining patients, sepsis, active alcoholism and relapse of chronic viral hepatitis are the most common reported precipitating factors. An excessive systemic inflammatory response seems to play a crucial role in the development of ACLF. Using a liver-adapted sequential organ assessment failure score, it is possible to triage and prognosticate the outcome of patients with ACLF. The course of ACLF is dynamic and changes over the course of hospital admission. Most of the patients will have a clear prognosis between day 3 and 7 of hospital admission and clinical decisions such as evaluation for liver transplant or discussion over goals of care could be tailored using clinical scores. Bioartificial liver support systems, granulocyte-colony stimulating factors or stem-cell transplant are in the horizon of medical care of this patient population; however, data are too premature to implement them as standard of care.


A novel add-on collimator for preclinical radiotherapy applications using a standard cell irradiator: design, construction, and validation.
Preclinical radiotherapy applications require dedicated irradiation systems which are expensive and not widely available. In this work, a clinical dual source 137 Cs cell irradiator was adapted to deliver 1-cm diameter preclinical treatment beams using a lead and stainless steel custom-made collimator to treat one or two mice at a time. The dosimetric characteristics of all the different components of the system (including collimator, phantoms, and radiation sources) have been simulated with EGSnrc Monte Carlo methods. The collimator was constructed based on these simulations and the calculated results were verified against dosimetric measurements with MOSKin detectors, GAFchromic films, and dosimetric gels. The comparisons showed an agreement, in terms of full width half maximum values, between the simulated and the measured dose cross profiles at the midline within 4% for both gel dosimetry and GAFchromic films. Out of beam dose, measured in air at the collimator midplane with MOSFET detectors was between 6% and 10% of the beam axis dose. The dimensions of the beam are constant along the vertical axis of the collimator and also the simulated and measured Percentage Depth Dose (PDD) curves show an agreement within 1%. The collimator design developed in this work allows the creation of a beam with the necessary characteristics for ablative radiotherapy treatments on small animals using a standard clinical cell irradiator. This collimator design will make advanced preclinical studies with ablative beams possible for all those institutions which do not have collimated preclinical irradiators available.


Magnetic resonance spectroscopy of cerebral cortex is normal in hereditary hyperekplexia due to mutations in the GLRA1 gene.
Excessive startling and stiffness in hereditary hyperekplexia has been attributed to lack of inhibition at either the cortical or brainstem level. Six patients with hereditary hyperekplexia (HH) and a confirmed mutation in the gene encoding the alpha(1) subunit of the glycine receptor (GLRA1) underwent single voxel (1)H magnetic resonance spectroscopy (MRS) of the brainstem and an area of frontal cortex and white matter using a method that allows absolute quantification of metabolites. The results of MRS were within normal limits, although there was a tendency for the neuronal marker N-acetyl aspartate to be reduced in the brainstem of patients compared with that in controls. Thus, we found no evidence to support a deficit in the cerebral cortex in patients with hereditary hyperekplexia due to mutations in the GLRA1 gene.


A Novel NOD2-associated Mutation and Variant Blau Syndrome: Phenotype and Molecular Analysis.
To describe the clinical and molecular implications of a novel mutation in the NOD2/CARD15 gene on a family and its seven affected members. We reviewed the clinical presentations of family members who came to our center for refractory uveitis. Genetic testing and molecular testing was performed. All affected members had adult onset recurrent non-granulomatous panuveitis. The inheritance pattern suggested an autosomal dominant disease and genetic analysis identified a novel mutation in the NOD2 gene that converted amino acid 600 from glutamate to alanine (E600A). Transfection of the E600A NOD2 into human embryonic kidney-293 (HEK293) cells revealed constitutive activation and a reduced ability to respond to the NOD2 ligand, muramyl dipeptide (MDP) as compared with wild-type NOD2. The E600A mutation in the NOD2 gene may confer a higher penetrance of uveitis but a later onset of milder forms of non-ocular involvement.


Gender disparities in clozapine prescription in a cohort of treatment-resistant schizophrenia in the South London and Maudsley case register.
Gender disparities in treatment are apparent across many areas of healthcare. There has been little research into whether clozapine prescription, the first-line treatment for treatment-resistant schizophrenia (TRS), is affected by patient gender. This retrospective cohort study identified 2244 patients with TRS within the South London and Maudsley NHS Trust, by using a bespoke method validated against a gold-standard, manually coded, dataset of TRS cases. The outcome and exposures were identified from the free-text using natural language processing applications (including machine learning and rules-based approaches) and from information entered in structured fields. Multivariable logistic regression was carried out to calculate the odds ratios for clozapine prescription according to patients' gender, and adjusting for numerous potential confounders including sociodemographic, clinical (e.g., psychiatric comorbidities and substance use), neutropenia, functional factors (e.g., problems with occupation), and clinical monitoring. Clozapine was prescribed to 77% of the women and 85% of the men with TRS. Women had reduced odds of being prescribed clozapine as compared to men after adjusting for all factors included in the present study (adjusted OR: 0.66; 95% CI 0.44-0.97; p = 0.037). Women with TRS are less likely to be prescribed clozapine than men with TRS, even when considering the effects of multiple clinical and functional factors. This finding suggests there could be gender bias in clozapine prescription, which carries ramifications for the relatively poorer care of women with TRS regarding many outcomes such as increased hospitalisation, mortality, and poorer quality of life.


Mirna Expression in Bladder Cancer and Their Potential Role in Clinical Practice.
To date more than 3000 miRNA sequences have been described in humans and registered at miRBase since their discovery. However, the functions of only a few of these miRNAs have been experimentally determined using deep sequencing technology. Aberrant miRNA expression has been associated with differentiation, invasion and metastasis in several cancers. In this context, recent reports have suggested that miRNAs play important roles in the regulation of target genes by binding to complementary regions of messenger transcripts to repress their translation or regulate degradation. In addition, the expression profiles of certain miRNAs can function biologically as oncogenes and tumor suppressor genes. In this review, we summarize the relationship between miRNAs expression and Bladder Cancer (BC). A comprehensive review of the literature has shown a differential expression between malignant and normal tissues, and that miRNAs could be the driving molecules of the BC progression. Similarly, the expression levels of miRNAs in urine and blood samples of BC patients have been demonstrated to be different from healthy people, a finding that might have diagnostic value. In conclusion, the understanding of miRNAs mechanisms and cell distribution provides new opportunities for diagnosis, prognostic, disease monitoring and personalized therapy of BC patients.


Oxadiazolone derivatives, new promising multi-target inhibitors against M. tuberculosis.
A set of 19 oxadiazolone (OX) derivatives have been investigated for their antimycobacterial activity against two pathogenic slow-growing mycobacteria, Mycobacterium marinum and Mycobacterium bovis BCG, and the avirulent Mycobacterium tuberculosis (M. tb) mc26230. The encouraging minimal inhibitory concentrations (MIC) values obtained prompted us to test them against virulent M. tb H37Rv growth either in broth medium or inside macrophages. The OX compounds displayed a diversity of action and were found to act either on extracellular M. tb growth only with moderated MIC50, or both intracellularly on infected macrophages as well as extracellularly on bacterial growth. Of interest, all OX derivatives exhibited very low toxicity towards host macrophages. Among the six potential OXs identified, HPOX, a selective inhibitor of extracellular M. tb growth, was selected and further used in a competitive labelling/enrichment assay against the activity-based probe Desthiobiotin-FP, in order to identify its putative target(s). This approach, combined with mass spectrometry, identified 18 potential candidates, all being serine or cysteine enzymes involved in M. tb lipid metabolism and/or in cell wall biosynthesis. Among them, Ag85A, CaeA, TesA, KasA and MetA have been reported as essential for in vitro growth of M. tb and/or its survival and persistence inside macrophages. Overall, our findings support the assumption that OX derivatives may represent a novel class of multi-target inhibitors leading to the arrest of M. tb growth through a cumulative inhibition of a large number of Ser- and Cys-containing enzymes involved in various important physiological processes.


Independent Vector Analysis (IVA) for Group fMRI Processing of Subcortical Area.
During functional MRI (fMRI) studies, blood oxygenation-level dependent (BOLD) signal associated with neuronal activity acquired from multiple individuals are subject to the derivation of group-averaged brain activation patterns. Unlike other cortical areas, subcortical areas such as the thalamus and basal ganglia often manifest smaller, biphasic BOLD signal that are aberrant from signals originating from cortices. Independent component analysis (ICA) can offer session/individual specific brain activation maps without a priori assumptions regarding the timing or pattern of the signal responses. The small activation loci within the subcortical areas are sparsely distributed among the subjects, and a conventional group processing method based on the general linear model (GLM) or ICA may fail to characterize the activation loci. In this paper, we present an independent vector analysis (IVA) to overcome these limitations by offering an analysis of additional dependent components (compared to the ICA-based method) that are assigned for use in the automated grouping of dependent (i.e. similar) activation patterns across subjects. The proposed IVA algorithm was applied to simulated data, and its utility was confirmed from real fMRI data employing a trial-based hand motor task. A GLM and the group ICA of the fMRI toolbox (GIFT) were also applied for comparison. From the analysis of activation patterns within subcortical areas, in which the hemodynamic responses (HRs) often deviate from a canonical, model-driven HR, IVA detected task-related activation loci that were not detected through GLM and GIFT. IVA may offer a unique advantage for inferring group activation originating from subcortical areas.


[Severe form of Rotavirus infection during the period of seasonal rise of its incidence].
The authors describe the results of observation and clinico-laboratory examination of patients during seasonal rises of the rotavirus infection incidence in 5 districts of the Moscow region in 1984-1987. The diseases ran their courses in the form of gastroenteritis and enteritis and were marked by a great number of the grave patterns (42-46%), mainly in children of the first three years of life, by a high percentage (77.5%) of the aggravated premorbid status of patients with the grave patterns. The rotavirus nature of diarrheas was supported by the data of direct electron microscopy, ELISA, and solid-phase coagglutination test. The symptom-complex of the clinical manifestations of rotavirus infection was characterized in detail. The histologic and morphometric data pertaining to two lethal outcomes were discussed. It has been demonstrated that respiratory viral infections, primarily influenza, produced an adverse effect, promoting the formation of the severe patterns and the onset of unfavourable outcomes of rotavirus infection in children.


Prophylactic compared with therapeutic ibuprofen analgesia in first-trimester medical abortion: a randomized controlled trial.
To compare the effectiveness of two oral analgesic regimens in first-trimester medical abortion. We randomly assigned 250 participants undergoing first-trimester abortion with mifepristone and misoprostol at three clinics to two ibuprofen regimens: therapeutic (800 mg every 4-6 hours as needed for pain) or prophylactic (800 mg starting 1 hour before the misoprostol dose, then every 4-6 hours for 48 hours regardless of pain, then as needed). We asked each participant to record her maximum pain on a scale of 0-10 daily thereafter. Of participants assigned to the prophylactic and therapeutic regimens, 111 of 123 (90%) and 117 of 127 (92%), respectively, provided follow-up data. More than 80% of the participants in each group complied with their assigned treatment. Participants in the prophylactic group used substantially more ibuprofen than those in the therapeutic group (median of nine and four tablets, respectively). The mean maximum pain score was 7.1 in the prophylactic group and 7.3 in the therapeutic group (standard deviations 2.5 and 2.2, respectively); the difference was not statistically significant (P=.87, adjusted for site). Duration of pain, verbal pain ratings reported at follow-up, and use of other analgesics did not differ significantly by group (all P>.05). No significant benefit of the prophylactic regimen was apparent in any population subgroup. Abortion failure and ibuprofen side effects in the two groups were similar. We found no evidence that prophylactic administration of ibuprofen reduces pain severity or duration in first-trimester medical abortion. The average pain severity experienced by participants using both regimens was high. ClinicalTrials.gov, www.clinicaltrials.gov, NCT01457521. I.


Evidence that non-deletional mechanisms are responsible for inducing and maintaining unresponsiveness after intrathymic injection of non-professional antigen presenting cells.
Intrathymic inoculation of donor alloantigen and concomitant immunosuppressive treatment can induce immune unresponsiveness to alloantigen. To examine the role of non-deletional mechanisms in the development of unresponsiveness, fractionated splenocytes were injected into only 1 lobe of the thymus. Untreated CBA (H2(k)) mice or controls pre-treated with anti-CD4 monoclonal antibody alone (on Day -28 and -27 relative to transplantation) acutely rejected C57BL/10 (H2(b)) cardiac allografts. Intrathymic inoculation of unfractionated splenocytes, resting B (rB) cells, or dendritic cells into both thymic lobes with the antibody resulted in indefinite survival of cardiac allografts. In contrast, when donor rB cells or dendritic cells were delivered into a single lobe of the thymus with the antibody, only rB cells induced indefinite prolongation of graft survival; unfractionated splenocytes or dendritic cells were markedly less effective. Mice that had 1 of the 2 thymic lobes removed were able to reject grafts even when treated with the antibody 27 days before transplantation. Therefore, T-cell export from 1 thymic lobe was sufficient to induce graft rejection. Finally, adoptive transfer of splenocytes from mice with long-term surviving primary grafts resulting from the intrathymic injection of rB cells significantly prolonged a graft from the same donor strain in a naive syngeneic recipient. Taken together, these data suggest that regulatory mechanisms generated by intrathymic injection of a non-professional antigen presenting cell, in this study donor rB cells, suppressed the rejection response mediated by T cells exported from the uninjected lobe.


Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification.
Clinical studies with the Abl tyrosine kinase inhibitor STI-571 in chronic myeloid leukemia demonstrate that many patients with advanced stage disease respond initially but then relapse. Through biochemical and molecular analysis of clinical material, we find that drug resistance is associated with the reactivation of BCR-ABL signal transduction in all cases examined. In six of nine patients, resistance was associated with a single amino acid substitution in a threonine residue of the Abl kinase domain known to form a critical hydrogen bond with the drug. This substitution of threonine with isoleucine was sufficient to confer STI-571 resistance in a reconstitution experiment. In three patients, resistance was associated with progressive BCR-ABL gene amplification. These studies provide evidence that genetically complex cancers retain dependence on an initial oncogenic event and suggest a strategy for identifying inhibitors of STI-571 resistance.