Rett syndrome: review of biological abnormalities.
The Rett syndrome (RS) is a peculiar, sporadic, atrophic disorder, almost entirely confined to females. After the first six months of life there is developmental slowing with reduced communication and head growth for about one year. This is followed by a rapid destructive stage with severe dementia and loss of hand skills (with frequent hand wringing), apraxia and ataxia, autistic features and irregular breathing with hyperventilation. Seizures often supervene. Subsequently there is some stabilization in a pseudo-stationary stage during the preschool to school years, associated with more emotional contact but also abnormalities of the autonomic and skeletal systems. After the age of 15-20 years, a late motor deterioration occurs with dystonia and frequent spasticity but seizures become milder. RS has generally been considered an X-linked disorder in which affected females represent a new mutation, with male lethality. Linkage studies suggested a critical region at Xq28. In 1999, mutations in the gene MECP2 encoding X-linked methyl cytosine-binding protein 2 (MeCP2) were found in a proportion of Rett girls. This protein can bind methylated DNA. Analyses are leading to much further investigation of mutants and their effects on genes. Neuropathological and electrophysiological studies of RS are described. Description of neurometabolic factors includes reduced levels of dopamine, serotonin, noradrenaline and choline acetyltransferase (ChAT) in brain, also estimation of nerve growth factors, endorphin, substance P, glutamate and other amino acids and their receptor levels. The results of neuroimaging are surveyed, including volumetric magnetic resonance imaging (MRI) and positron emission tomography (PET).


The effectiveness and risks of fluoroscopically guided cervical transforaminal injections of steroids: a systematic review with comprehensive analysis of the published data.
To determine the effectiveness and risks of fluoroscopically guided cervical transforaminal injection of corticosteroids in the treatment of radicular pain. Systematic review of the literature with comprehensive analysis of the published data. Three reviewers with formal training in evidence-based medicine searched the literature on fluoroscopically guided cervical transforaminal injection of steroids (CTFIS). Each reviewer independently assessed the methodology of studies found and appraised the quality of the evidence presented. The primary outcome assessed was relief of radicular pain. Other outcomes such as reduction in surgery rate and complications were noted if reported. The evidence on each outcome was appraised in accordance with the Grades of Recommendation, Assessment, Development, and Evaluation (GRADE) system of evaluating evidence. The searches yielded 16 primary publications on effectiveness. Available evidence, derived mainly from observational studies, suggests that approximately 50% of patients experience 50% relief of radicular pain for at least 4 weeks after CTFIS, and the intervention may have surgery-sparing effects. The literature also contains 21 articles with primary reports of serious complications, including 13 deaths and many catastrophic neurological injuries. The evidence of pain-relieving effects, of surgery-sparing effects, and of risks of CTFIS were all rated as of very low quality according to the GRADE system. In patients with cervical radicular pain, fluoroscopically guided CTFIS may be effective in easing pain and reducing need for surgery. However, the evidence of effectiveness is of very low quality, and the benefits of the procedure are compromised by the risks of serious complications.


Identifying and correcting spatial bias in opportunistic citizen science data for wild ungulates in Norway.
Many publications make use of opportunistic data, such as citizen science observation data, to infer large-scale properties of species' distributions. However, the few publications that use opportunistic citizen science data to study animal ecology at a habitat level do so without accounting for spatial biases in opportunistic records or using methods that are difficult to generalize. In this study, we explore the biases that exist in opportunistic observations and suggest an approach to correct for them. We first examined the extent of the biases in opportunistic citizen science observations of three wild ungulate species in Norway by comparing them to data from GPS telemetry. We then quantified the extent of the biases by specifying a model of the biases. From the bias model, we sampled available locations within the species' home range. Along with opportunistic observations, we used the corrected availability locations to estimate a resource selection function (RSF). We tested this method with simulations and empirical datasets for the three species. We compared the results of our correction method to RSFs obtained using opportunistic observations without correction and to RSFs using GPS-telemetry data. Finally, we compared habitat suitability maps obtained using each of these models. Opportunistic observations are more affected by human access and visibility than locations derived from GPS telemetry. This has consequences for drawing inferences about species' ecology. Models naïvely using opportunistic observations in habitat-use studies can result in spurious inferences. However, sampling availability locations based on the spatial biases in opportunistic data improves the estimation of the species' RSFs and predicted habitat suitability maps in some cases. This study highlights the challenges and opportunities of using opportunistic observations in habitat-use studies. While our method is not foolproof it is a first step toward unlocking the potential of opportunistic citizen science data for habitat-use studies.


Further potential limitations of the undifferentiated faecal egg count reduction test for the detection of anthelmintic resistance in sheep.
Fifteen (36%) of 42 mixed gastro-intestinal nematode infections in sheep, identified as drench-susceptible by the undifferentiated faecal egg count reduction test, were found to harbour anthelmintic-resistant worms when analysed on the basis of changes in the egg counts of individual nematode genera. Most of these cases involved resistance in a single nematode genus, with Ostertagia and Trichostrongylus being implicated most frequently. The possible contributory role of larval culture results in helping to reduce the chances of the faecal egg count reduction test producing these and similar types of errors is discussed.


[Treatment of stroke by reducing the body temperature; 'Paracetamol (acetaminophen) in stroke' (PAIS): start of a clinical trial].
Subfebrile temperatures and fever in acute stroke are associated with poor functional outcome. A 1 degree C rise in body temperature may double the risk of a poor outcome in patients who are admitted within 12 hours from the onset of symptoms. Two randomised double-blind clinical trials in patients with acute ischaemic stroke have shown that treatment with a daily dose of 6 g paracetamol results in a small but rapid and potentially worthwhile reduction of 0.3 degree C (95% CI: 0.1-0.5) in body temperature. It has been hypothesized that early antipyretic therapy reduces the risk of death or dependency in patients with acute stroke, even if they are normothermic. For this reason, a multicentre, randomized, double-blind clinical trial comparing high-dose paracetamol with placebo in 2500 patients has been launched. This study has been named 'Paracetamol (acetaminophen) in stroke' (PAIS). The primary outcome is death or dependency at three months. The study protocol is simple, and the amount of data to be gathered is limited. The trial will run for four years.


Metabolic tuning of inhibition regulates hippocampal neurogenesis in the adult brain.
Hippocampus-engaged behaviors stimulate neurogenesis in the adult dentate gyrus by largely unknown means. To explore the underlying mechanisms, we used tetrode recording to analyze neuronal activity in the dentate gyrus of freely moving adult mice during hippocampus-engaged contextual exploration. We found that exploration induced an overall sustained increase in inhibitory neuron activity that was concomitant with decreased excitatory neuron activity. A mathematical model based on energy homeostasis in the dentate gyrus showed that enhanced inhibition and decreased excitation resulted in a similar increase in neurogenesis to that observed experimentally. To mechanistically investigate this sustained inhibitory regulation, we performed metabolomic and lipidomic profiling of the hippocampus during exploration. We found sustainably increased signaling of sphingosine-1-phosphate, a bioactive metabolite, during exploration. Furthermore, we found that sphingosine-1-phosphate signaling through its receptor 2 increased interneuron activity and thus mediated exploration-induced neurogenesis. Taken together, our findings point to a behavior-metabolism circuit pathway through which experience regulates adult hippocampal neurogenesis.


Allergic fetal priming leads to developmental, behavioral and neurobiological changes in mice.
The state of the mother's immune system during pregnancy has an important role in fetal development and disruptions in the balance of this system are associated with a range of neurologic, neuropsychiatric and neurodevelopmental disorders. Epidemiological and clinical reports reveal various clues that suggest a possible association between developmental neuropsychiatric disorders and family history of immune system dysfunction. Over the past three decades, analogous increases have been reported in both the incidence of neurodevelopmental disorders and immune-related disorders, particularly allergy and asthma, raising the question of whether allergic asthma and characteristics of various neurodevelopmental disorders share common causal links. We used a mouse model of maternal allergic asthma to test this novel hypothesis that early fetal priming with an allergenic exposure during gestation produces behavioral deficits in offspring. Mothers were primed with an exposure to ovalbumin (OVA) before pregnancy, then exposed to either aerosolized OVA or vehicle during gestation. Both male and female mice born to mothers exposed to aerosolized OVA during gestation exhibited altered developmental trajectories in weight and length, decreased sociability and increased marble-burying behavior. Moreover, offspring of OVA-exposed mothers were observed to have increased serotonin transporter protein levels in the cortex. These data demonstrate that behavioral and neurobiological effects can be elicited following early fetal priming with maternal allergic asthma and provide support that maternal allergic asthma may, in some cases, be a contributing factor to neurodevelopmental disorders.


Testing the stress-buffering effects of self engagement among soldiers on a military operation.
The present research examined whether self engagement moderated the stressor-strain relationship in a sample of U.S. soldiers deployed on a peacekeeping mission to Bosnia. Given that individuals were nested within groups (Army companies), we examined the relationships of interest using multilevel random coefficient models. The results of these analyses revealed that self engagement interacted with three different stressors (lower amounts of sleep, work stress, family stress) in the prediction of psychological distress. In support of self engagement as a buffer against stress, when stressors were low (high amounts of sleep, low levels of work stress, and family stress), soldiers reported relatively low levels of psychological distress and physical symptoms, regardless of whether they were engaged in or disengaged from their job. However, when stressor levels were high, soldiers who were engaged in their job reported less elevation in reports of psychological distress than soldiers who were disengaged from their job. The results are discussed in terms of the potential mechanisms by which self engagement might protect individuals from stressful circumstances.


The Interdisciplinary Generalist Curriculum (IGC) project: an overview of its experience and outcomes.
The Interdisciplinary Generalist Curriculum (IGC) Project was a competitive, seven-year demonstration project funded by the Health Resources and Services Administration (HRSA). It was established to determine whether specific interdisciplinary innovations in preclinical medical school curricula could affect students' selection of careers in family medicine, general internal medicine, or general pediatrics. Through collaboration among the three generalist disciplines, the IGC innovation exposed all preclinical students in ten demonstration schools to a new or significantly enhanced preclinical curriculum that included a direct supervised clinical experience with a generalist physician preceptor. The project was managed by an interdisciplinary executive committee that was codirected by one representative each from family medicine, general internal medicine, and general pediatrics. A national advisory committee with representation from the academic and professional organizations of family medicine, internal medicine, pediatrics, and osteopathy provided input to the executive committee in guiding the project. The project was externally evaluated. Major outcomes of the IGC Project include sustained curricular changes in ten institutions, prompted by relatively few dollars and demonstration of models for collaboration at institutional and national levels. This supplement describes the IGC Project's experience and outcomes so that others may draw pertinent information to apply to their own efforts in medical education.


Hericium erinaceus extracts alter behavioral rhythm in mice.
Hericium erinaceus (HE), an edible mushroom, has been used as a herbal medicine in several Asian countries since ancient times. HE has potential as a medicine for the treatment and prevention of dementia, a disorder closely linked with circadian rhythm. This study investigated the effects of the intake of HE extracts on behavioral rhythm, photosensitivity of the circadian clock, and clock gene mRNA expression in the suprachiasmatic nucleus (SCN), a central clock, in mice. Although the HE ethanol extract only affected the offset time of activity, the HE water extract advanced the sleep-wake cycle without affecting the free-running period, photosensitivity, or the clock gene mRNA expression in SCN. In addition, both extracts decreased wakefulness around end of active phase. The findings of the present study suggest that HE may serve as a functional food in the prevention and treatment of Alzheimer's disease and delayed sleep phase syndrome.


Results after surgery for severe Dupuytren's contracture: does a dynamic extension splint influence outcome?
Seventy-six consecutive patients were operated on for advanced Dupuytren's contracture and the results evaluated after nine months with special reference to the use of a dynamic extension splint. The patients were separated into three groups: those in whom the splint was used according to our guidelines (n = 15); those in whom the splint was used, but inadequately (n = 15); and those who did not require splinting (n = 24). Our results nine months postoperatively were similar to those of other studies in showing that the fifth proximal interphalangeal joint constituted the greatest problem. Comparison of the three groups indicated that splinting the way we used it did not influence the natural course of the disease after operation.


Regulation of surfactant-like particle secretion by Caco-2 cells.
Surfactant-like particle (SLP) is a phosphatidylcholine (PC)-rich membrane produced in the small intestine, and its secretion is increased by fat feeding. In Caco-2 cells known to produce SLP, preincubation with [(3)H]palmitate labelled the SLP and was used as a marker for newly secreted membrane. SLP-associated PC and protein (d=1.07-1.08 g/ml in a linear non-equilibrium NaBr gradient) were secreted in parallel with triacylglycerols (TG) and at a rate about twice the control rate in response to feeding cells with an oleate/egg PC mixture. Cholesterol and apolipoprotein A-I identified only a small peak corresponding to high-density lipoprotein (HDL), but the largest peak corresponded with SLP (d=1.07-1.08). Palmitate incorporation into PC showed a similar small peak migrating at the density of HDL, but most labelled PC secreted from the cells was due to SLP. PC secretion, alkaline phosphatase activity, and newly synthesized immunoprecipitated SLP proteins from conditioned serum-free media migrated together at a density of >/=1.21 g/ml in a lipoprotein NaBr step gradient, and represented SLP. Glycerol incorporated into TG migrated at a peak density of 1.12 g/ml, consistent with HDL secretion from cells incubated in serum-free media. These data confirm that the secreted PC in SLP is distinct from lipoprotein particles. Incorporation of [(3)H]palmitate into the PC fraction of either whole cell homogenate or isolated brush border membranes was not affected by oleate/egg PC feeding. Both Pluronic L-81, an inhibitor of chylomicron secretion, and BMS-197636-02, a microsomal triglyceride transfer protein inhibitor, blocked the secretion of both TG and PC. Elevation of intracellular cAMP levels that stimulate surfactant secretion from type II pneumocytes caused a 50% reduction in SLP and TG secretion from Caco-2 cells. These results confirm the SLP response to fat feeding found in vivo, further supporting a role for SLP in TG secretion from the enterocyte, and show that the regulation of SLP secretion differs from that of pulmonary surfactant.


Mixed Effects Modeling Using Stochastic Differential Equations: Illustrated by Pharmacokinetic Data of Nicotinic Acid in Obese Zucker Rats.
Inclusion of stochastic differential equations in mixed effects models provides means to quantify and distinguish three sources of variability in data. In addition to the two commonly encountered sources, measurement error and interindividual variability, we also consider uncertainty in the dynamical model itself. To this end, we extend the ordinary differential equation setting used in nonlinear mixed effects models to include stochastic differential equations. The approximate population likelihood is derived using the first-order conditional estimation with interaction method and extended Kalman filtering. To illustrate the application of the stochastic differential mixed effects model, two pharmacokinetic models are considered. First, we use a stochastic one-compartmental model with first-order input and nonlinear elimination to generate synthetic data in a simulated study. We show that by using the proposed method, the three sources of variability can be successfully separated. If the stochastic part is neglected, the parameter estimates become biased, and the measurement error variance is significantly overestimated. Second, we consider an extension to a stochastic pharmacokinetic model in a preclinical study of nicotinic acid kinetics in obese Zucker rats. The parameter estimates are compared between a deterministic and a stochastic NiAc disposition model, respectively. Discrepancies between model predictions and observations, previously described as measurement noise only, are now separated into a comparatively lower level of measurement noise and a significant uncertainty in model dynamics. These examples demonstrate that stochastic differential mixed effects models are useful tools for identifying incomplete or inaccurate model dynamics and for reducing potential bias in parameter estimates due to such model deficiencies.


Unfreezing unspent social special-purpose funds for the Covid-19 crisis: Critical reflections from India.
In India, the government launched a US$22.6 billion financial support package for the poor and marginalized as a result of Covid-19. Approximately US$ 4.2 billion (INR 310 billion) came from a vast pile of unspent social special-purpose funds. How and why did such a large volume of funds accumulate in the first place, and why did it take a public health emergency to release them? What might be the consequences of their use under such emergency conditions - especially for our understanding of governance and accountability in social welfare provision? This paper presents a brief analysis of two preliminary case studies of specific social special-purpose funds in India. We rely on a handful of unstructured interviews and informal discussions with subnational government officials, civil society actors, trade union representatives, and local community leaders that began in January 2020, and which were pursued virtually following the lockdown. This is bolstered by analysis of primary documents, including Comptroller and Auditor General of India (CAG) reports, relevant laws, and contemporary press coverage. We argue that non-disbursement should be understood as a institutional matter, and not only as technical or implementation failure. Moreover, as such funds are likely to mushroom following Covid-19, our findings suggest that policymakers should focus on the institutional design, decision-making and accountability structures for the flow and distribution of Covid funds, rather than merely emphasising their collection.


ROC curve analysis for biomarkers based on pooled assessments.
Interleukin-6 is a biomarker of inflammation which has been suggested as having potential discriminatory ability for myocardial infarction. Because of its high assaying cost it is very expensive to evaluate this marker. In order to reduce this cost we propose pooling the specimens. In this paper we examine the efficiency of ROC curve analysis, specifically the estimation of the area under the ROC curve, when dealing with pooled data. We study the effect of pooling when there are only a fixed number of individuals available for testing and pooling is carried out to save on the number of assays. Alternatively we examine how many pooled assays of size g are necessary to provide essentially the same information as N individual assays. We measure loss of information by means of the change in root mean square error of the estimate of the area under the ROC curve and study the extent of this loss via a simulation study.


Brain and peripheral metabolism of 5-hydroxytryptophan-14C following peripheral decarboxylase inhibition.
Following selective peripheral decarboxylase inhibition, a greater fraction of peripherally measured monoamine metabolites should derive from central nervous system monoamine metabolism. In rats pretreated with the peripheral decarboxylase inhibitor L-alpha-(3,4-dihydroxybenzyl)-alpha-hydrazinopropionic acid (MK-486) (50 mg/kg i.p. or i.v.), the peripheral formation of 5-hydroxytryptamine-14C (5-HT-14C) from the precursor DL-5-hydroxytryptophan-14C (5-HTP-14C) was reduced by 82 to 100% over a 4-hour interval. There was a marked increase in the penetration of 5-HTP-14C into brain and in the appearance in brain of 5-HT-14C and 5-hydroxyindoleacetic acid-14C (5-HIAA-14C). The cerebral metabolism of 5-HTP-14C in vehicle-pretreated animals appeared to occur mainly in the brain vascular compartment. Following MK-486 pretreatment, the in vivo metabolism of 14C-5-hydroxyindoles in the extracerebral compartment obeys linear kinetics as does the uptake and decarboxylation of 5-HTP-14C in brains. The cerebral turnover of 5-HT-14C appeared to increase as a function of the dose of 5-HTP injected. This phenomenon may be a result of either saturation of brain 5-HT storage sites or nonspecific metabolism of 5-HTP in brain catecholamine neurons. A highly signigificant correlation was found between the brain and blood 5-HIAA-14C levels following MK-486. The adrenal formation of 5-HT, which was not inhibited by MK-486, did not appear to contribute significantly to the blood 5-HIAA-14c levels. These results suggest the possibility that following peripheral decarboxylase inhibition a larger fraction of peripherally measured 5-HIAA may derive from central nervous system 5-HT metabolism.


A Delphi study to determine the European core curriculum for Master programmes in genetic counselling.
Genetic counsellors have been working in some European countries for at least 30 years. Although there are great disparities between the numbers, education, practice and acceptance of these professionals across Europe, it is evident that genetic counsellors and genetic nurses in Europe are working autonomously within teams to deliver patient care. The aim of this study was to use the Delphi research method to develop a core curriculum to guide the educational preparation of these professionals in Europe. The Delphi method enables the researcher to utilise the views and opinions of a group of recognised experts in the field of study; this study consisted of four phases. Phases 1 and 4 consisted of expert workshops, whereas data were collected in phases 2 and 3 (n=35) via online surveys. All participants in the study were considered experts in the field of genetic counselling. The topics considered essential for genetic counsellor training have been organised under the following headings: (1) counselling; (2) psychological issues; (3) medical genetics; (4) human genetics; (5) ethics, law and sociology; (6) professional practice; and (7) education and research. Each topic includes the knowledge, skills and attitudes required to enable genetic counsellors to develop competence. In addition, it was considered by the experts that clinical practice should comprise 50% of the educational programme. The core Master programme curriculum will enable current courses to be assessed and inform the design of future educational programmes for European genetic counsellors.


The Tzanck smear in the diagnosis of cutaneous herpes simplex.
We examined the effectiveness of the Tzanck preparation compared with viral isolation in 32 lesions of cutaneous herpes simplex infection in 30 consecutive patients. Seventy-eight percent had positive cultures, even though the clinical history and lesion morphological findings were typical in all situations. Other authors have also noted that viral isolation does not reach 100% even in the face of typical clinical history. Fifty-three percent of the Tzanck smears were positive. These preparations had a high correlation with positive viral cultures (94.1%). Vesicles were more likely to yield a positive culture than Tzanck smear (100% and 66.7%, respectively). Pustules yielded positive cultures in 72.8% and positive Tzanck smears in 54.5%, while crusted-ulcer lesions were usually not positive by either technique (34%, 16%). A positive Tzanck preparation is an accurate, rapid tool for the diagnosis of cutaneous herpes simplex.


Charcoal haemoperfusion and haemodialysis in acute intermittent porphyria.
Charcoal haemoperfusion has been advocated as a means of removing delta aminolaevulinic acid, which accumulates in attacks of acute intermittent porphyria. A woman presented with acute intermittent porphyria unresponsive to conventional treatment and with pain that was difficult to control. Charcoal haemoperfusion was performed in series with haemodialysis for two hours daily on four consecutive days. Although during this treatment serum and urinary concentrations of delta aminolaevulinic acid and porphobilinogen were considerably reduced, they had returned to pretreatment values 24 hours after the end of treatment. Abdominal pain was not relieved. Although a longer course of treatment might have had a more favourable outcome, this seems unlikely in view of the rapid rebound of serum concentration of delta amino-laevulinic acid after each haemoperfusion.


Correlation between the numbers of gammadelta T cells and CD4+ HLA-DR+ T cells in broncho-alveolar lavage fluid from patients with diffuse lung disease.
CD4+ HLA-DR+ T cells are known to be increasing in broncho-alveolar lavage fluid (BALF) from patients with sarcoidosis, and related to disease activity. Although there are several reports that the number of gammadelta T cells in peripheral blood from patients with sarcoidosis are increasing, contradictory assertions can be seen about the number of gammadelta T cells in BALF, and the clinical significance on the presence of gammadelta T cells in disease site of patients with diffuse lung disease including sarcoidosis. The absolute number of gammadelta T cells and CD4+ HLA-DR+ T cells in BALF were determined by flow cytometry in 107 patients with diffuse lung diseases; 56 with sarcoidosis, 36 with collagen vascular diseases with lung involvement and 15 with idiopathic pulmonary fibrosis. We also measured the number of the transferrin receptor-positive macrophages in BALF. The correlation between gammadelta T cells and activated (maybe antigen-specific) T cells and macrophages were evaluated. Sarcoidosis patients were also evaluated from the data of the number of gammadelta T cells in peripheral blood by flow cytometry and clinical backgrounds. A significant correlation between the numbers of these two cell types was detected in each of the three patient groups. The percentage of peripheral gammadelta T cells was markedly increased in 7 sarcoidosis patients, each of whom also showed affected organs other than lung, however, 5 individuals did not show an increased number of gammadelta T cells in BALF. The number of gammadelta T cells in BALF did not correlate with the number of transferrin receptor-positive macrophages in all three patient groups. These results suggest that the increased number of gammadelta T cells in diffuse lung diseases likely plays a role in immunosurveillance and contributes to the activation of antigen-specific alphabeta T cell.


Behavioral sensitization to ethanol is modulated by environmental conditions, but is not associated with cross-sensitization to allopregnanolone or pentobarbital in DBA/2J mice.
The ability of ethanol to facilitate GABA(A) receptor-mediated transmission may result in GABA(A) receptor alterations during repeated ethanol administration, and lead to dynamic behavioral changes, including sensitization to the locomotor stimulant effect of ethanol. Since alterations in GABA(A) receptors are likely to alter sensitivity to GABAergic drugs such as 3alpha-hydroxy-5alpha-pregnan-20-one (allopregnanolone) and pentobarbital, we determined whether enhanced sensitivity to ethanol was associated with enhanced sensitivity (cross-sensitization) to these drugs. Two procedures that produced differences in the magnitude of expression of ethanol-induced locomotor sensitization were used. After habituation to testing procedures for 2 days, female DBA/2J mice were injected with ethanol or saline for 12 days. On the following day, locomotion was recorded after a challenge injection of ethanol (2 g/kg), allopregnanolone (10 or 17 mg/kg), or pentobarbital (10 or 20 mg/kg). Due to evidence that exposure to the test chambers influenced sensitization, in some experiments, mice were exposed to the test apparatus on the day prior to challenge. Exposure to the test apparatus prior to drug challenge attenuated the expression of ethanol sensitization, compared with mice without this pre-exposure. Cross-sensitization was not observed to either allopregnanolone or pentobarbital under any condition; however, some groups of repeated ethanol-treated mice displayed tolerance to the initial stimulant effects of allopregnanolone and pentobarbital. These studies indicate that behavioral sensitization to ethanol is not associated with cross-sensitization to pentobarbital or allopregnanolone, and that the expression of ethanol sensitization is influenced by the relative novelty of the test chamber. In addition, these results do not support a mechanism in which alterations in the neurosteroid or barbiturate modulatory sites of the GABA(A) receptor are responsible for the expression of sensitization to the locomotor stimulant effects of ethanol.


Thrombolysis and fibrinolytic parameters during heparin treatment of deep vein thrombosis.
Thirty-nine patients with symptomatic, venographically verified deep vein thrombosis (DVT) were studied during treatment with heparin in order to investigate the correlation between the venographic changes and parameters of heparin therapy or fibrinolytic components. Venograms were scored with a 40-grade scale, and after one week a significant improvement with an average reduction of the thrombi of 17% was observed. No statistically significant correlation was found between reduction of thrombus size and duration of heparin treatment, total amount of heparin administered, mean levels of APTT, plasmin-alpha 2-antiplasmin complex (PAP), tissue plasminogen activator (t-PA) antigen or t-PA-inhibitor. Only a short history of the thrombus was significantly correlated to thrombolysis. The concentrations of PAP and t-PA-inhibitor were not influenced, while that of t-PA antigen showed a significant increase during heparin infusion. Even if statistically significant correlations were not obtained, the patients with pronounced thrombolytic effect had high PAP-levels. Furthermore, patients with high t-PA-inhibitor levels had no lysis. The results suggest, that also other factors than plasminogen dependent fibrinolysis are of importance for the thrombolytic effect.


Glucose flux in relation to energy expenditure in malnourished patients with and without cancer during periods of fasting and feeding.
Glucose dynamics, energy metabolism, and nitrogen balance were studied in eight malnourished cancer patients and seven malnourished patients without cancer. Glucose flux was measured by single injection of [6-3H]glucose and [U-14C]glucose. Energy expenditure was measured by indirect calorimetry. Each patient was studied after an overnight fast and during constant gastric infusion of a formula diet. Cancer patients had elevated glucose flux when fasting, corresponding to 42% of their spontaneous daily intake of glucose. At least one-half of the elevated flux in cancer patients compared with controls was due to increased recycling of glucose carbon after an overnight fast. Feeding doubled the total glucose flux in both cancer and control patients. The recycling was unchanged in the cancer group and disappeared in the controls during feeding. The increased glucose flux in cancer patients was concomitant with normal resting energy expenditure during periods of both fasting and feeding. Glucose flux in relation to energy expenditure was doubled in cancer patients compared to controls, and the glucose flux in fed cancer patients was similar to the rate of glucose infusion, which shows that the endogenous production of glucose was not inhibited. Cancer and control patients reached a comparable positive energy and nitrogen balance, allowing for their overall caloric intake. Our results show that cancer patients seem to have a characteristically increased glucose demand, which contributes to their weight loss when fasting. The energy drain by this elevated glucose flux can explain, as a maximum estimate, a loss of about 0.9 kg of body fat per 30-day period.


Carry-over effects of the membrane interface probe.
The membrane interface probe (MIP) is widely used to characterize the subsurface distribution of volatile organic compounds (VOCs). One problem that arises during MIP application is that disproportionately high MIP signals are obtained after passing source zones which contain mobile or residual phases. This serious problem occurs because of a carry-over effect, in particular caused by compound-specific retention times in the conventional unheated transfer line, commonly used during such an investigation. The objective of this study was to perform a qualitative methodical field evaluation of the carry-over effect of a conventional MIP system with a conventional unheated transfer line. This was achieved by coupling a mobile mass spectrometer to the MIP device. Results obtained were then further compared with those achieved using a laser induced fluorescence (LIF) system. Because of this coupling, time- and depth-dependent signals for different substances became known. Field evaluation data obtained showed complex superpositions of compounds with MIP system results. As a result of this superposition, MIP signals from the saturated zone beneath the source zone (zone with free and/or residual phase) are blurred and are therefore not representative of particular depths. However, utilizing multidirectional probing alongside conventional MIP probing (forwards and backwards), it was possible to detect the upper and lower phase boundary of the source zone. These MIP results correlated excellently with the LIF results. An important conclusion that can be drawn from the field investigation is that coupling a mobile mass spectrometer to the MIP system enables advanced MIP signal interpretation to be successfully achieved.


A Hybrid Framework for Intrusion Detection in Healthcare Systems Using Deep Learning.
The unbounded increase in network traffic and user data has made it difficult for network intrusion detection systems to be abreast and perform well. Intrusion Systems are crucial in e-healthcare since the patients' medical records should be kept highly secure, confidential, and accurate. Any change in the actual patient data can lead to errors in the diagnosis and treatment. Most of the existing artificial intelligence-based systems are trained on outdated intrusion detection repositories, which can produce more false positives and require retraining the algorithm from scratch to support new attacks. These processes also make it challenging to secure patient records in medical systems as the intrusion detection mechanisms can become frequently obsolete. This paper proposes a hybrid framework using Deep Learning named "ImmuneNet" to recognize the latest intrusion attacks and defend healthcare data. The proposed framework uses multiple feature engineering processes, oversampling methods to improve class balance, and hyper-parameter optimization techniques to achieve high accuracy and performance. The architecture contains <1 million parameters, making it lightweight, fast, and IoT-friendly, suitable for deploying the IDS on medical devices and healthcare systems. The performance of ImmuneNet was benchmarked against several other machine learning algorithms on the Canadian Institute for Cybersecurity's Intrusion Detection System 2017, 2018, and Bell DNS 2021 datasets which contain extensive real-time and latest cyber attack data. Out of all the experiments, ImmuneNet performed the best on the CIC Bell DNS 2021 dataset with about 99.19% accuracy, 99.22% precision, 99.19% recall, and 99.2% ROC-AUC scores, which are comparatively better and up-to-date than other existing approaches in classifying between requests that are normal, intrusion, and other cyber attacks.


Cultural perceptions and practices around menarche and adolescent menstruation in the United States.
This overview details the persistence of negative characterizations of the menstrual cycle as a feature of the current cultural context in which girls begin menstrual life in the United States. In addition, research on girls' current menstrual attitudes and experiences within this context are reviewed. Current research suggests that girls are not very knowledgeable about menstruation, and that menstrual education continues to provide girls with mixed messages, such as: menstruation is a normal, natural event, but it should be hidden. Girls' attitudes and expectations about menstruation are negatively biased and have been found to contribute to self-objectification, body shame, and lack of agency in sexual decision-making. Although preparation has been found to be associated with more positive menarcheal and menstrual experiences, specificity about what constitutes "good" preparation has not been well articulated. Implications for promoting the menstrual cycle among young girls as a vital sign in service of monitoring their health are discussed.


Reversal of nostril dominance by posture.
Reflex reversal of differential air flow through nostrils (DAFTN) by the adoption of suitable lateral recumbent position was studied in male Nigerian subjects. The subjects who demonstrated more air flow through the left nostril initially in the supine position, adopted left lateral recumbent (LLR) position which brought the decongested left nostril to the down-side and the congested right nostril to the upside. Within 3 to 4 minutes after the adoption of LLR position, the air flow through the nostrils was equalised and by 11th minute the DAFTN was reversed, with more air flowing through the upsided right nostril. Return of the subjects to supine position could not result in the recovery to the initial pattern of DAFTN. It is suggested that reversal of DAFTN with adoption of suitable lateral recumbent position was a reflex effect due to the pressure stimuli to the lower lateral side of the body particularly around the shoulder region and effect was mediated through the sympathetic innervation to mucosal venous sinuses of the nostrils. Possibility of gravitational effect was ruled out.


[Relapse in Hodgkin lymphoma].
The retrospective study revealed the 15% relapse rate in patients with stage II-IV unfavorable prognosis Hodgkin lymphoma, 5-year OS in relapsed patients was 84%. Karnofsky score less than 80 (p=0,0001), more than 1 extranodal lesion (p=0,0004), extensive (equal to stage III-IV) involvement on relapse (p=0,001), b-symptoms on relapse (p=0,023), more than 5 lymph nodal lesions (p=0,027), albumin level less than 40 g per liter (p=0,037), detection of new nodal lesions (p=0,041) were shown by discriminative analysis as the therapy effect predictors in patients with Hodgkin lymphoma relapse. In patients with second-line therapy failure the actuarial survival rate was lower by 55% in comparison to patients with chemosensitive relapses (40% and 95%). The secondary therapy resistance was shown to be an unfavorable prognosis predictive factor (p=0,0001). The multifactorial overall survival analysis revealed the following adverse prognostic factors: failure of second-line treatment (p=0,0001), first early relapse (p=0,01), albumin level on relapse less than 40 g per liter (p=0,02), use of standard chemotherapy instead of irradiation (p=0,02). The relapse patients with 1 or less risk factors had 95% 5-year OS, the patients with 3 or more adverse risk factors had 70% OS (p=0,0002). The lowest 10-year OS was observed in patients with 2 or more adverse risk factors, 48% and 28% accordingly. Adverse risk factors must be considered while choosing the optimal treatment strategy aimed at better survival rate.


Ternary alkali ion thiogallates, A5GaS4 (A = Li and Na), with isolated tetrahedral building units and their ionic conductivities.
Two new ternary thiogallates in the A5GaS4 (A = Li (i) and Na (ii)) series have been synthesized for the first time employing a gas passing route using oxide precursors and a high temperature solid state route using stoichiometric combinations of elements, respectively. Li5GaS4 crystallizes in the P21/m space group and the structure is built up of layers of corner sharing tetrahedra of LiS4 and GaS4 stacked along the a-axis and the octahedrally coordinated Li ions residing in the interlayer space. Na5GaS4 crystallizes in the Pbca space group and the structure consists of isolated (GaS4)5- tetrahedra held together by charge balancing sodium ions in distorted tetrahedral and octahedral coordination geometries. Measurements of ionic conductivity of the compounds showed room temperature ionic conductivities of 1.8 × 10-7 and 4.0 × 10-7 S cm-1 with activation energies of 0.54 and 0.28 eV, respectively, for I and II. Density functional theory calculations show close agreement in structural parameters with the measured data and predict band gaps of 2.75 eV (I) and 2.70 eV (II). Single point hybrid functional calculations result in band gaps of 3.95 and 3.65 eV correspondingly, in better agreement with the experimental value of ∼4.1 eV for both. Bond valence energy landscape maps suggest the absence of any suitable diffusion path for Li in Li5GaS4. On the other hand, BVEL maps of Na5GaS4 confirm that the tetrahedrally coordinated Na ions are responsible for ionic conduction, whereas the involvement of octahedrally coordinated Na ions in the conduction process could not be discerned.


Anatomical model of the human trunk for analysis of respiratory muscles mechanics.
A realistic two-dimensional (2D) model of the human trunk was developed for quantitative analysis of the relative contribution to breathing mechanics of seven groups of respiratory muscles. Along with noninvasive measurements of electromyography (EMG) signals from respiratory muscles near the skin surface, it provides predictions for the forces generated by inner respiratory muscles as well as the instantaneous work of each muscle. The results revealed that inspiratory muscles reach their maximal force towards the end of inspiration, while expiratory muscles reach their maximal force at mid-expiration. Inspiratory muscles contributed to the work of breathing even at low efforts, while that of the expiratory muscles was observed only at relatively high efforts. The study clearly showed that the diaphragm muscle generates forces, which are of the same order as those generated by other inspiratory muscles, but performed 60-80% of the inspiratory work. The contribution of the external intercostal muscle to inspiration was not negligible, especially at high breathing efforts.


Identification of an essential proximal sequence element in the promoter of the telomerase RNA gene of Tetrahymena thermophila.
Telomerase is a ribonucleoprotein reverse transcriptase that synthesizes and maintains telomeric DNA. Studies of telomeres and telomerase are facilitated by the large number of linear DNA molecules found in ciliated protozoa, such as Tetrahymena thermophila. To examine the expression of telomerase, we investigated the transcription of the RNA polymerase III-directed gene encoding the RNA subunit (TER1) of this enzyme. A chimeric gene containing the Glaucoma chattoni TER1 transcribed region flanked by 5' and 3' Tetrahymena regions was used to identify promoter elements following transformation of Tetrahymena cells. Disruption of a conserved proximal sequence element (PSE) located at -55 in the Tetrahymena TER1 5' flanking region eliminated expression of the chimeric gene. In addition, mutation of an A/T-rich element at -25 decreased expression markedly. A gel mobility shift assay and protein-DNA cross-linking identified a PSE-binding polypeptide of 50-60 kDa in Tetrahymena extracts. Gel filtration analysis revealed a native molecular mass of approximately 160 kDa for this binding activity. Our results point to a similar architecture between ciliate telomerase RNA and metazoan U6 small nuclear RNA promoters.


Radiologically Isolated Syndrome: 10-Year Risk Estimate of a Clinical Event.
We have previously identified male sex, younger age, and the presence of spinal cord lesions as independent factors that increase the 5-year risk for evolution from radiologically isolated syndrome (RIS) to multiple sclerosis. Here, we investigate risk factors for the development of a clinical event using a 10-year, multinational, retrospectively identified RIS dataset. RIS subjects were identified according to 2009 RIS criteria and followed longitudinally as part of a worldwide cohort study. We analyzed data from 21 individual databases from 5 different countries. Associations between clinical and magnetic resonance imaging (MRI) characteristics and the risk of developing a first clinical event were determined using multivariate Cox regression models. Additional follow-up data were available in 277 of 451 RIS subjects (86% female). The mean age at RIS diagnosis was 37.2 years (range, 11-74 years), with a median clinical follow-up of 6.7 years. The cumulative probability of a first clinical event at 10 years was 51.2%. Age, positive cerebrospinal fluid for oligoclonal bands, infratentorial lesions on MRI, and spinal cord lesions, were baseline independent predictors associated with a subsequent clinical event. The presence of gadolinium-enhanced lesions during follow-up was also associated with the risk of a seminal event. The reason for MRI and gadolinium-enhancing lesions at baseline did not influence the risk of a subsequent clinical event. Approximately half of all individuals with RIS experience a first clinical event within 10 years of the index MRI. The identification of independent predictors of risk for symptom onset may guide education and clinical management of individuals with RIS. ANN NEUROL 2020;88:407-417.


Preclinical Activity of Sacituzumab Govitecan, an Antibody-Drug Conjugate Targeting Trophoblast Cell-Surface Antigen 2 (Trop-2) Linked to the Active Metabolite of Irinotecan (SN-38), in Ovarian Cancer.
Background: Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy. Sacituzumab govitecan (SG) is a novel antibody-drug-conjugate (ADC) targeting trophoblast-antigen-2 (Trop-2), a cell surface glycoprotein highly expressed in many epithelial tumors, to deliver SN-38, the active metabolite of irinotecan. This study aimed to evaluate Trop-2 expression in EOC tissues and the preclinical activity of SG against primary EOC cell lines and xenografts. Methods: Trop-2 expression was assessed in 90 formalin-fixed-paraffin-embedded tumors and nine primary tumor cell lines by immunohistochemistry (IHC) and flow cytometry, respectively. Trop-2 expression and cell viability after exposure to SG in primary tumor cell lines, non-targeting control ADC, and SG-parental antibody hRS7 were evaluated using flow-cytometry-based-assays. Antibody-dependent-cell-cytotoxicity (ADCC) against Trop-2+ and Trop-2- EOC cell lines was tested in vitro using 4 h Chromium-release-assays. In vivo activity of SG was evaluated against Trop-2+ EOC xenografts. Results: Moderate-to-strong staining was seen in 47% (42/90) of ovarian tumors by IHC while 89% (8/9) of the primary EOC cell lines overexpressed Trop-2 by flow cytometry. EOC Trop-2+ were significantly more sensitive to SG compared to control ADC (p < 0.05). Both SG and hRS7 mediated high ADCC activity against Trop-2+ cell lines. SG also induced significant bystander killing of Trop-2- tumor cells admixed with Trop-2+ EOC cells. In in vivo experiments SG treatment demonstrated impressive anti-tumor activity against chemotherapy-resistant EOC xenografts. Conclusion: SG demonstrates remarkable preclinical activity against biologically aggressive and chemotherapy-resistant EOC cell lines and a significant bystander effect against EOC cell lines with heterogenous Trop-2 expression. Clinical trials are warranted.


Nutrient Recommendations for Cancer Patients Treated with Pelvic Radiotherapy, with or without Comorbidities.
Radiotherapy is one of the main treatment options used in pelvic cancers. Ionizing radiation induces damage to surrounding tissues, resulting in disruption of normal physiological functions and symptoms such as diarrhea, tenesmus, incontinence, and rectal bleeding, which can all significantly alter the patient's quality of life. These patients are at increased risk of developing protein-calorie malnutrition and micronutrient deficiencies. Therefore, designing a proper nutritional intervention plan, with an optimal proportion of protein, fat, and carbohydrates, is required to reduce or even reverse the patients' poor nutritional status, increase their tolerance and response to oncology treatment, decrease the rate of complications and improve their quality of life. The aim of this review was to establish a nutritional plan that includes recommendations on macronutrient proportions and micronutrient intake in patients receiving pelvic radiotherapy. The following nutritional plan has been recommended in the literature: Energy: 28-31 kcal/kg/day, using the Harris-Benedict formula adjusted for body weight in obese patients; protein: 20-30%; fat: 30-40%; and carbohydrates: 40-50%. The maintenance of adequate levels of Vitamin D, Vitamin E, Vitamin A, calcium, magnesium, thiamin, riboflavin, and niacin must be emphasized. Physical activity must also be increased to maintain muscle mass. Nutrient requirements must be established in an integral manner, considering the patient's age, nutritional status, and the presence of comorbidities. Unnecessary dietary restrictions should be avoided to ensure an adequate nutritional status.


Desmoplastic small round cell tumor: evaluation of reverse transcription-polymerase chain reaction and fluorescence in situ hybridization as ancillary molecular diagnostic techniques.
Desmoplastic small round cell tumor (DSRCT) is a rare, biologically aggressive soft tissue neoplasm of uncertain differentiation, most often arising in the abdominal and pelvic cavities of adolescents and young adults with a striking male predominance. Histologically, it is characterized by islands of uniform small round cells in prominent desmoplastic stroma, and it has a polyimmunophenotypic profile, typically expressing WT1 and cytokeratin, desmin, and neural/neuroendocrine differentiation markers to varying degrees. Tumors at other sites and with variant morphology are more rarely described. DSRCT is associated with a recurrent t(11;22)(p13;q12) translocation, leading to the characteristic EWSR1-WT1 gene fusion. Fluorescence in situ hybridization (FISH), to detect EWSR1 rearrangement, and reverse transcription-polymerase chain reaction (RT-PCR) to assess for EWSR1-WT1 fusion transcripts are routine diagnostic ancillary tools. We present a large institutional comparative series of FISH and RT-PCR for DSRCT diagnosis. Twenty-six specimens (from 25 patients) histologically diagnosed as DSRCT were assessed for EWSR1 rearrangement and EWSR1-WT1 fusion transcripts. Of these 26 specimens, 24 yielded positive results with either FISH or RT-PCR or both. FISH was performed in 23 samples, with EWSR1 rearrangement seen in 21 (91.3%). RT-PCR was performed in 18 samples, of which 13 (72.2%) harbored EWSR1-WT1 fusion transcripts. The sensitivity of FISH in detecting DSRCT was 91.3%, and that of RT-PCR was 92.8% following omission of four technical failures. Therefore, both methods are comparable in terms of sensitivity. FISH is more sensitive if technical failures for RT-PCR are taken into account, and RT-PCR is more specific in confirming DSRCT. Both methods complement each other by confirming cases that the other method may not. In isolation, FISH is a relatively non-specific diagnostic adjunct due to the number of different neoplasms that can harbor EWSR1 rearrangement, such as Ewing sarcoma. However, in cases with appropriate morphology and a typical pattern of immunostaining, FISH is confirmatory of the diagnosis.


Gender-specific differences in the central nervous system's response to anesthesia.
Males and females are physiologically distinct in their responses to various anesthetic agents. The brain and central nervous system (CNS), the main target of anesthesia, are sexually dimorphic from birth and continue to differentiate throughout life. Accordingly, gender has a substantial impact on the influence of various anesthetic agents in the brain and CNS. Given the vast differences in the male and female CNS, it is surprising to find that females are often excluded from basic and clinical research studies of anesthesia. In animal research, males are typically studied to avoid the complication of breeding, pregnancy, and hormonal changes in females. In clinical studies, females are also excluded for the variations that occur in the reproductive cycle. Being that approximately half of the surgical population is female, the exclusion of females in anesthesia-related research studies leaves a huge knowledge gap in the literature. In this review, we examine the reported sex-specific differences in the central nervous system's response to anesthesia. Furthermore, we suggest that anesthesia researchers perform experiments on both sexes to further evaluate such differences. We believe a key goal of research studying the interaction of the brain and anesthesia should include the search for knowledge of sex-specific mechanisms that will improve anesthetic care and management in both sexes.


Transglutaminases as targets for pharmacological inhibition.
Transglutaminases (TGases), a family of enzymes that catalyze the formation of epsilon-(gamma-glutamyl)lysine isopeptide linkage, play an important physiological role in hemostasis, wound healing, assembly and remodeling of the extracellular matrix, cell signaling and apoptosis. Although many members of this class of enzymes have been known for decades, their role in various physiological and pathological processes is still a subject of substantial research and debate. Convincing evidence exists that TGases are involved in formation of cytotoxic proteinatious aggregates in Alzheimer's, Huntington's and other neurodegenerative diseases. However, it is not clear if elevated levels of TGases play a causative or protective role in several of these processes. Increased or defective TGase activity is a factor in cortical cataract formation, lamellar ichtyosis and fibrosis. TGase creates epitopes for the production of autoantibodies in celiac disease and possibly other autoimmune diseases. Another TGase, Factor XIIIa, is involved in the etiology of vascular diseases. Modulation of TGase activity through its selective inhibition may have therapeutic benefit in a wide variety of diseases. This paper will examine TGases as targets for the development of new therapeutics and review the progress in discovery of selective inhibitors of these enzymes.


A UWB Radar Signal Processing Platform for Real-Time Human Respiratory Feature Extraction Based on Four-Segment Linear Waveform Model.
This paper presents an ultra-wideband (UWB) impulse-radio radar signal processing platform used to analyze human respiratory features. Conventional radar systems used in human detection only analyze human respiration rates or the response of a target. However, additional respiratory signal information is available that has not been explored using radar detection. The authors previously proposed a modified raised cosine waveform (MRCW) respiration model and an iterative correlation search algorithm that could acquire additional respiratory features such as the inspiration and expiration speeds, respiration intensity, and respiration holding ratio. To realize real-time respiratory feature extraction by using the proposed UWB signal processing platform, this paper proposes a new four-segment linear waveform (FSLW) respiration model. This model offers a superior fit to the measured respiration signal compared with the MRCW model and decreases the computational complexity of feature extraction. In addition, an early-terminated iterative correlation search algorithm is presented, substantially decreasing the computational complexity and yielding negligible performance degradation. These extracted features can be considered the compressed signals used to decrease the amount of data storage required for use in long-term medical monitoring systems and can also be used in clinical diagnosis. The proposed respiratory feature extraction algorithm was designed and implemented using the proposed UWB radar signal processing platform including a radar front-end chip and an FPGA chip. The proposed radar system can detect human respiration rates at 0.1 to 1 Hz and facilitates the real-time analysis of the respiratory features of each respiration period.


Crystallisation in oxide glasses - a tutorial review.
Glasses and glass-ceramics have had a tremendous impact upon society and continue to have profound industrial, commercial and domestic importance. A remarkable number of materials, with exceptional optical and mechanical properties, have been developed and enhanced using the glass-ceramic method over many years. In order to develop glass-ceramics, glass is initially prepared via high temperature synthesis and subsequently heat treated, following a carefully designed and controlled process. A glass-ceramic system comprises crystalline and non-crystalline phases; in multicomponent systems these phases are significantly different from the initial glass composition. The properties of glass-ceramics are defined by microstructure, crystal morphology as well as the final chemical composition and physical properties of the residual glass. Knowing the mechanism of glass crystallisation, it is possible to predict and design a glass-ceramic system with near-ideal properties that exactly fulfil the requirements for a particular application. This tutorial review is a basic introduction to the crystallisation in glasses and mainly focuses on silicate and closely related oxide glasses. The review describes and discusses key learning points in five different sections, which facilitate the understanding of glass crystallisation and development of glass-ceramics.


Investigation of common genetic risk factors between thyroid traits and breast cancer.
Breast cancer (bc) risk is suspected to be linked to thyroid disorders, however observational studies exploring the association between bc and thyroid disorders gave conflicting results. We proposed an alternative approach by investigating the shared genetic risk factors between bc and several thyroid traits. We report a positive genetic correlation between bc and thyroxine (FT4) levels (corr = 0.13, p-value = 2.0x10-4) and a negative genetic correlation between bc and thyroid-stimulating hormone (TSH) levels (corr = -0.09, p-value = 0.03). These associations are more striking when restricting the analysis to estrogen receptor-positive bc. Moreover, the polygenic risk scores (PRS) for FT4 and hyperthyroidism are positively associated to bc risk (OR = 1.07, 95%CI: 1.00-1.13, p-value = 2.8x10-2 and OR = 1.04, 95%CI: 1.00-1.08, p-value = 3.8x10-2, respectively), while the PRS for TSH is inversely associated to bc risk (OR = 0.93, 95%CI: 0.89-0.97, p-value = 2.0x10-3). Using the PLACO method, we detected 49 loci associated to both bc and thyroid traits (p-value<5x10-8), in the vicinity of 130 genes. An additional colocalization and gene-set enrichment analyses showed a convincing causal role for a known pleiotropic locus at 2q35 and revealed an additional one at 8q22.1 associated to both bc and thyroid cancer. We also found two new pleiotropic loci at 14q32.33 and 17q21.31 that were associated to both TSH levels and bc risk. Enrichment analyses and evidence of regulatory signals also highlighted brain tissues and immune system as candidates for obtaining associations between bc and TSH levels. Overall, our study sheds light on the complex interplay between bc and thyroid traits and provides evidence of shared genetic risk between those conditions.


Modelling of particle paths passing through an ultrasonic standing wave.
Within an ultrasonic standing wave particles experience acoustic radiation forces causing agglomeration at the nodal planes of the wave. The technique can be used to agglomerate, suspend, or manipulate particles within a flow. To control agglomeration rate it is important to balance forces on the particles and, in the case where a fluid/particle mix flows across the applied acoustic field, it is also necessary to optimise fluid flow rate. To investigate the acoustic and fluid forces in such a system a particle model has been developed, extending an earlier model used to characterise the 1-dimensional field in a layered resonator. In order to simulate fluid drag forces, CFD software has been used to determine the velocity profile of the fluid/particle mix passing through the acoustic device. The profile is then incorporated into a MATLAB model. Based on particle force components, a numerical approach has been used to determine particle paths. Using particle coordinates, both particle concentration across the fluid channel and concentration through multiple outlets are calculated. Such an approach has been used to analyse the operation of a microfluidic flow-through separator, which uses a half wavelength standing wave across the main channel of the device. This causes particles to converge near the axial plane of the channel, delivering high and low particle concentrated flow through two outlets, respectively. By extending the model to analyse particle separation over a frequency range, it is possible to identify the resonant frequencies of the device and associated separation performance. This approach will also be used to improve the geometric design of the microengineered fluid channels, where the particle model can determine the limiting fluid flow rate for separation to occur, the value of which is then applied to a CFD model of the device geometry.


Convergent transcription confers a bistable switch in Enterococcus faecalis conjugation.
Convergent gene pairs with head-to-head configurations are widespread in both eukaryotic and prokaryotic genomes and are speculated to be involved in gene regulation. Here we present a unique mechanism of gene regulation due to convergent transcription from the antagonistic prgX/prgQ operon in Enterococcus faecalis controlling conjugative transfer of the antibiotic resistance plasmid pCF10 from donor cells to recipient cells. Using mathematical modeling and experimentation, we demonstrate that convergent transcription in the prgX/prgQ operon endows the system with the properties of a robust genetic switch through premature termination of elongating transcripts due to collisions between RNA polymerases (RNAPs) transcribing from opposite directions and antisense regulation between complementary counter-transcripts. Evidence is provided for the presence of truncated RNAs resulting from convergent transcription from both the promoters that are capable of sense-antisense interactions. A mathematical model predicts that both RNAP collision and antisense regulation are essential for a robust bistable switch behavior in the control of conjugation initiation by prgX/prgQ operons. Moreover, given that convergent transcription is conserved across species, the mechanism of coupling RNAP collision and antisense interaction is likely to have a significant regulatory role in gene expression.


Rabid dog illegally imported to France from Morocco, August 2011.
In August 2011, a case of canine rabies was notified to the French veterinary services. The dog was a three-month-old puppy illegally imported from Morocco that presented behavioural changes on 1 August and was admitted to a veterinary clinic on 6 August. It died the following day and the body was shortly sent to the national reference centre where rabies was laboratory-confirmed on 11 August. Contact tracing and post-exposure treatment were initiated immediately.


Effects of insulin exposure upon the metabolism of rat aortic media: influence of hydrostatic forces.
When added in vitro, insulin exerts little or no effect on the metabolism of arterial media. However, indirect evidence suggests that an in vivo exposure of arterial tissue to insulin stimulates its metabolism. To evaluate the possibility that hemodynamic forces existing only in vivo may account for this difference, we studied the effect of brief (9-minute) exposures to insulin of isolated rat thoracic aorta perfused in situ at low (34 mm Hg = 4.6 kPa) and high (70 mm Hg = 9.4 kPa) static pressures. After perfusion, incorporation of glucose-derived 14C into CO2 and lipids, and production of lactate by incubated aortic tissue, were measured. At low perfusion pressure, insulin did not affect the metabolism of the media but strongly stimulated all aspects of the glucose metabolism of the adventitia. At high pressure, insulin again stimulated the adventitia but now also significantly stimulated (+66%) the synthesis of glucose-derived lipids by the media; production of CO2 and lactate by the media did not increase significantly. These results indicate that insulin directly stimulates at least one pathway of glucose metabolism by the media; and that hemodynamic forces are an important factor in determining the response of aortic media to insulin.


Trivalent lanthanide interactions with a terdentate bis(dialkyltriazinyl)pyridine ligand studied by electrospray ionization mass spectrometry.
The 2,6-bis(5,6-dialkyl-1,2,4-triazin-3-yl)pyridines (DATPs) belong to a new family of extracting agents recently developed in the framework of nuclear fuel reprocessing. These molecules exhibit exceptional properties to separate actinides(III) from lanthanides(III) in nitric acid solutions. A previous work showed that electrospray ionization mass spectrometry (ESI-MS) is a reliable technique to provide solution data such as stoichiometries and conditional stability constants of various DATP complexes with europium and evidenced the unusual capability of DiPTP [bis(di-iso-propyltriazinyl)pyridine] ligand to form 1:3 complexes in nitric acid solution. This latter result is further investigated by considering DiPTP complexation features with the complete lanthanide family. As a starting point of the experimental procedure used for stability constant evaluation, the intensity distribution of ions detected by ESI-MS is studied for solutions containing Ln(NO(3))(3) in water/methanol (1:1 v/v) with the pH value set at 2.8 and 4.6 by HNO(3) additions. At pH 2.8, the nitrate anions are found to prevent lanthanides from processes occurring within the ion source: redox phenomena or gas-phase reactions with methanol which give species such as [Ln(MeO)(2)](+). Thus, the total intensity of MS signals from [Ln(NO(3))(2)(H(2)O)(p)(MeOH)(n)](+) ions is found proportional to the metal ion concentration. At pH 4.6, with lower nitrate concentration, the nature of the species identified on mass spectra depends on the electronic properties of the lanthanide elements. It is shown that Ln(III) complexation with DiPTP leads to the exclusive formation of 1:3 complexes with the whole lanthanide series which may be due not only to the hydrophobic exterior of the ligand but also to the unusual electronic density distribution in DATP ligands as compared with other aza-aromatic ligands. The conditional stability constants of the 1:3 lanthanide(III) complexes with DiPTP have been determined at pH 2.8 and are found to increase almost regularly from La (log beta(3)(app) = 11.7 +/- 0.1) to Lu (log beta(3)(app) = 16.7 +/- 0.8). Moreover, the kinetic stability of the gas-phase 1:3 complexes obtained by electrospray has been investigated by energy-resolved collision-induced dissociation and provides useful information on the bonding and structure.


Household training vs. mass campaigns: a better method of health communication for preventing malaria.
Malaria is endemic in several states of India with high tribal population. Effective use of long-lasting insecticide treated bed nets (LLITNs) can reduce the burden of malaria in these settings. This study assessed the knowledge and behaviour regarding malaria in a tribal population and compared two health education strategies for enhancing effective utilisation of bed nets. A community-based intervention study was carried out among 218 households in two villages. One community received household level training on effective utilisation of LLITNs while the others received training in a mass campaign and outcomes were measured after 2 months. The study showed that the proportion utilising LLITNs was significantly higher among those receiving household level training as compared to those who received the mass campaign. Household level training appears to be a more effective form of health education for improving knowledge and promoting use of LLITNs in these isolated community groups.


Cluster headache and lifestyle habits.
Cluster headache (CH) has traditionally been associated with certain anthropometric features, personality traits, and lifestyle features. This article focuses on lifestyle features in patients with CH. Especially excessive smoking and alcohol consumption have been ascribed to patients with CH. Despite country-specific habits and a time trend, smoking is much more prevalent among CH patients compared with the general population. Although excessive alcohol consumption was reported in early studies, this was not corroborated more recently. On the contrary, patients with CH seem to avoid alcohol, particularly during active phases, likely due to its ability to trigger attacks. Present studies are purely descriptive. Thus, the associations sketched give no information about the long-term effects of smoking or alcohol consumption on the course of CH.


Direct measurements of structural forces and twist transitions in cholesteric liquid crystal films with a surface force apparatus.
Using a surface force apparatus, a cholesteric liquid crystal was confined between two crossed cylindrical surfaces that induced strong planar anchoring and normal alignment of the chiral helix. The film thickness and total twist angle of the chiral molecular structure were simultaneously measured using multiple-beam optical interference. As the film thickness was increased and the chiral structure deformed, the twist angle remained almost unchanged until discontinuous changes occurred at critical distances that were equally spaced by one cholesteric half-pitch length. Structural deformations generated oscillatory elastic forces with periodically spaced maxima corresponding to twist transitions. These findings were reproduced using an equilibrium model of cholesteric confinement and force generation. The analysis indicates that the strength of the azimuthal surface anchoring on mica is high, exceeding 0.2 mJ m-2.


Spinal p38 activity and analgesic effect after low- and high-intensity electroacupuncture stimulation in a plantar incision rat model.
Postoperative pain is a major problem. Electroacupuncture (EA) has been accepted as a useful and low-risk complementary therapy for post-operative pain. Animal studies indicate that surgical incision activates p38 MAPK in the spinal microglia, which critically contributes to post-incisional nociceptive development. How EA affects incision-induced p38 activation is important but yet to be fully elucidated. Male adult rats received plantar incision (PI) at the right hind paw followed by 30-min EA of 4-Hz, one of two intensities (3 and 10mA), and at right ST36 (Zusanli) acupoint immediately after PI and for 3 successive days. EA analgesia was evaluated by von Frey fibers and Hargreaves' tests. Spinal p38 activation was examined by immunostaining. In separate groups, SB203580, a p38 inhibitor, was intrathecally injected alone or with EA to test the combining effect on nociception and spinal phospho-p38. EA of 10-mA significantly ameliorated mechanical allodynia, but 3-mA did not. None of them altered thermal hyperalgesia. Repeated EA could not inhibit phospho-p38 in the PI rats, contrarily, EA per se significantly induced phospho-p38 in the normal rats. Intrathecal SB203580 injection dose-dependently prevented PI-induced allodynia. Combination of low-dose SB203580 and 3-mA EA, which were ineffective individually, profoundly reduce post-PI allodynia. We demonstrated that 10-mA EA exerts a significant inhibition against post-PI mechanical hypersensitivity via a p38-independent pathway. Importantly, co-treatment with low-dose p38 inhibitor and 3-mA EA can counteract spinal phospho-p38 to exert strong analgesic effect. Our finding suggests a novel strategy to improve EA analgesic quality.


T-DNA from Agrobacterium Ti plasmid is in the nuclear DNA fraction of crown gall tumor cells.
The crown gall teratoma tumor line BT37, incited by Agrobacterium tumefaciens strain T37, has been found to contain part of the tumor-inducing plasmid, pTi T37, of the inciting strain. This foreign DNA segment, called T-DNA, is maintained at several copies per diploid tumor cell. We have examined subcellular DNA fractions from this tumor line in an effort to determine whether T-DNA is in chloroplasts, mitochondria, or nuclei. Tumor cell chloroplast DNA exhibited EcoRI and Bst I endonuclease cleavage patterns identical to those of normal tobacco chloroplast DNA. Tumor cell mitochondrial DNA exhibited a complex Bst I cleavage pattern that did not differ detectably from that of normal tobacco mitochondrial DNA. Southern blots of tumor chloroplast and mitochondrial cleavage products did not hybridize with labeled pTi T37 DNA, whereas blots of tumor cell nuclear DNA cleavage products hybridized strongly. We conclude that T-DNA is located not in chloroplasts or mitochondria but rather in the nuclear fraction of this tumor line.


Egg transmission of egg drop syndrome 1976 virus in fowl.
Egg transmission of egg drop syndrome 1976 virus (BC14 virus) in fowl was demonstrated in the second and third week after experimental infection. Eggs of BC14 virus infected hens were incubated weekly after disinfection with formaline gas. After 18 days of incubation, eggs with live embryos were homogenized. This egg material was fed to adult hens, housed in isolators. Seroconversion in these birds demonstrated egg transmission. It is suggested that egg transmission occurs as a result of viremia.


Hemoglobin Okaloosa (beta 48 (CD7) leucine leads to arginine). An unstable variant with low oxygen affinity.
A slow-moving hemoglobin with electrophoretic mobility similar to that of hemoglobin S was discovered in a white laboratory technologist. She had an elevated reticulocyte count, as did several members of her family. Her red cell survival was shortened. Amino acid analysis indicated that leucine at position beta48 (CD7) had been replaced by arginine. The abnormal hemoglobin, called Okaloosa, was heat-precipitable and had decreased oxygen affinity. It exhibited a greater change in oxygen affinity than hemoglobin A when 2,3 DPG was added to "stripped" hemolysates. These findings cannot be readily explained by current views of structure-function relationships in the hemoglobin molecule. However, it is of interest that the amino acid in position CD7 is normally leucine in the alpha, beta, delta, and gamma-hemoglobin chains and in the myoglobin chain of man and a wide variety of other vertebrates.


Anaerobic digestion of gelatinous water at laboratory and pilot scale and nitrogen inhibition.
The anaerobic digestion of the liquid residue (gelatinous water) coming from the production of fat from animal residue, was studied at laboratory and pilot scale. Biodegradability (>98%) and biogas potential (675 mL of biogas/g of COD(applied)) of this wastewater are very high. However, due to the high content on nitrogen, an inhibition of the anaerobic activity was observed for quite low concentrations of N-NH(3). Dilution of the wastewater and pH regulation in the reactor around 7.3 are the 2 solutions which were investigated to overcome the nitrogen inhibition at industrial scale. These two solutions were validated at laboratory scale in an anaerobic SBR and then onsite at pilot scale in a continuous reactor. A stable anaerobic digestion was observed in both reactors showing that no nitrogen inhibition was obtained when N-NH(3) concentration in the reactor was kept low.


Risk factors for catheter-related bloodstream infection: a prospective multicenter study in Brazilian intensive care units.
Central venous catheters (CVC) are devices of great importance in health care. The advantages gained from the use of catheters outweigh the complications that might result from their use, among which bloodstream infections (BSI). In spite of its importance, few national studies have addressed this issue. The aim this study was to determine the incidence of BSI in patients with CVC, hospitalized in ICU, as well as the variables associated with this complication. Multicentric cohort study carried out at ICUs of three hospitals at Universidade Federal de São Paulo complex. A total of 118 cases of BSI in 11.546 catheters day were observed: 10.22 BSI per 1,000 catheters day. On average, BSI was associated to seven additional days of hospital stay in our study (p < 0.001), with a significant difference between types of catheters. Concerning the place of insertion, there was no statistical difference in BSI rates. We concluded that a patient who uses a catheter for longer than 13 days presents a progressive risk for infection of approximately three times higher in relation to a patient who uses the catheter for less than 13 days (p < 0.001). The median duration of catheter use was 14 days among patients with BSI and 9 days in patients without infection (p < 0.001). There was higher prevalence of gram-negative infections. The risk factors for BSI were utilization of multiple-lumen catheters, duration of catheterization and ICU length of stay.


The potential role of homocysteine mediated DNA methylation and associated epigenetic changes in abdominal aortic aneurysm formation.
Previous studies have suggested that homocysteine (Hcy) has wide-ranging biological effects, including accelerating atherosclerosis, impairing post injury endothelial repair and function, deregulating lipid metabolism and inducing thrombosis. However, the biochemical basis by which hyperhomocysteinemia (HHcy) contributes to cardiovascular diseases (CVDs) remains largely unknown. Several case-control studies have reported an association between HHcy and the presence of abdominal aortic aneurysms (AAA) and there are supportive data from animal models. Genotypic data concerning the association between variants of genes involved in the methionine cycle and AAA are conflicting probably due to problems such as reverse causality and confounding. The multifactorial nature of AAA suggests the involvement of additional epigenetic factors in disease formation. Elevated Hcy levels have been previously linked to altered DNA methylation levels in various diseases. Folate or vitamin B12 based methods of lowering Hcy have had disappointingly limited effects in reducing CVD events. One possible reason for the limited efficacy of such therapy is that they have failed to reverse epigenetic changes induced by HHcy. It is possible that individuals with HHcy have an "Hcy memory effect" due to epigenetic alterations which continue to promote progression of cardiovascular complications even after Hcy levels are lowered. It is possible that deleterious effect of prior, extended exposure to elevated Hcy concentrations have long-lasting effects on target organs and genes, hence underestimating the benefit of Hcy lowering therapies in CVD patients. Therapies targeting the epigenetic machinery as well as lowering circulating Hcy concentrations may have a more efficacious effect in reducing the incidence of cardiovascular complications.


Acute appendicitis during pregnancy. Diagnosis and management.
Twelve patients underwent appendectomy during pregnancy or in the puerperium. The clinical presentation of acute appendicitis is altered during gestation, and diagnosis becomes increasingly difficult when close to term. Abdominal pain, nausea, and vomiting are important symptoms. Peritoneal signs occur in the right lower quadrant early in pregnancy, but the upper quadrant or entire right side are more common locations, as the appendix is displaced upward by the enlarging uterus. Delay in treatment is common because of uncertainty in making the diagnosis and hesitancy to proceed with surgery. In the group of six patients with perforation, there was one maternal death and a loss of three fetuses. There were no complications in the absence of perforation. Prompt diagnosis is the cornerstone of a good outcome, and early surgical intervention is indicated if acute appendicitis is suspected. Pregnancy is not a reason to delay surgery. We review the literature on this topic and present and analyze principles of management.


Supporting family carers of people with dementia: A discrete choice experiment of public preferences.
Community-based care for people with dementia is mainly provided by family carers, many of whom experience decreased mental, physical and financial well-being due to their caring role. Many countries are now implementing ageing-in-place policies that have increased pressure on family carers as care-work is redistributed from residential to community-based settings. Family caring responsibilities for people with dementia are made even more complicated by the economic, social and cultural expectations that underpin existing provision. Support for family carers is, therefore, an important policy topic across many countries. The focus of this paper is on the propensity of citizens to support enhanced care for family carers in Ireland, as demonstrated by their willingness-to-pay additional taxation to fund different combinations of carer support measures, developed through careful and sustained dialogue with multiple stakeholders, especially family carers themselves. We carried out a discrete choice experiment (DCE) with 509 members of the general public in Ireland between January and February 2021. Using mixed logit models, our findings show that citizens value four key attributes: regular caring breaks for family carers (day-care and long-break respite); financial compensation (carer's allowance); and emotional support (carer peer support groups). We also estimated the welfare impact of moving from current provision to enhanced support packages for family carers of people with dementia. The welfare gains accumulate to €1035.80 for the most complete levels of provision across the four support attributes. Overall, respondents in this paper showed empathy and understanding towards family carers of people with dementia through their willingness to contribute to funding additional services and supports.


One hundred useful references in pediatric cardiac intensive care: the 2012 update.
The specialty of pediatric cardiac critical care has undergone rapid scientific and clinical growth in the last 25 years. The Board of Directors of the Pediatric Cardiac Intensive Care Society assembled an updated list of sentinel references focused on the critical care of children with congenital and acquired heart disease. We encouraged board members to select articles that have influenced and informed their current practice or helped to establish the standard of care. The objective of this article is to provide clinicians with a compilation and brief summary of these updated 100 useful references. The list of 'One Hundred Useful References for Pediatric Cardiac Intensive Care' (2004) and relevant literature to the practice of cardiac intensive care. A subset of Pediatric Cardiac Intensive Care Society board members compiled the initial list of useful references in 2004, which served as the basis of the new updated list. Suggestions for relevant articles were submitted by the Pediatric Cardiac Intensive Care Society board members and selected pediatric cardiac intensivists with an interest in this project following the Society's meeting in 2010. Articles were considered for inclusion if they were named in the original list from 2004 or were suggested by Pediatric Cardiac Intensive Care Society board members and published before December 31, 2011. Following submission of the complete list by the Pediatric Cardiac Intensive Care Society board and contributing Society members, articles were complied by the two co-first authors (D.A., D.K.). The authors also performed Medline searches to ensure comprehensive inclusion of all relevant articles. The final list was then submitted to the Pediatric Cardiac Intensive Care Society board members, who ranked each publication. Rankings were compiled and the top 100 articles with the highest scores were selected for inclusion in this publication. The two co-first authors (D.A., D.K.) reviewed all existing summaries and developed summaries of the newly submitted articles. An updated compilation of 100 useful references for the critical care of children with congenital and acquired heart disease has been compiled and summarized here. Clinicians and trainees may wish to use this document as a reference for education in this complex and challenging subspecialty.


Virtual bladder tumor transurethral resection: an objective evaluation tool to overcome learning curves with and without photodynamic diagnostics.
Virtual reality (VR) simulation has been known to shorten learning curves. The aim of this research was to determine whether VR transurethral bladder tumor resection (TURBT) with and without photodynamic diagnostic (PDD) can improve endourological skills in novice and trained endoscopists. Residents without experience (n = 15) completed five 5-min white light VR-TURBT scenarios. The trained endoscopists (≥25 TURBTs; n = 15) performed five 5-min VR resections using white light and PDD cystoscopy. The VR-TURBT performance with the Uro-Trainer® (Karl Storz GmbH, Germany) was compared between the first and last 5 cases using computer-generated parameters [inspected area (%), resected tumor mass (%) and bleeding (ml)]. The trained endoscopists' parameters were compared with white light versus PDD-VR-TURBT. Among novices, the inspected area increased from 36.8 (±12.9) to 54.3% (±7.3), p < 0.05. Significant improvements were noted for resection rates from 26.5 (±9.6) to 52.0% (±6.0), p < 0.05. Trained urologists showed superior inspection [52.2 (±3.4) vs. 62.7% (±4.3), p = 0.003] and resection rates [43.8 (±3.3) vs. 57.1% (±5.3), p = 0.002] with PDD. The opportunity to evaluate objective patient-independent measurements establishes the Uro-Trainer as a beneficial tool in enhancing TURBT quality. Results demonstrated that novices improved their ability to perform complete resection, and the TURBT rates of trained urologists were superior in PDD compared with a white light setting.


Rhodobacter vinaykumarii sp. nov., a marine phototrophic alphaproteobacterium from tidal waters, and emended description of the genus Rhodobacter.
A rod-shaped, phototrophic, purple non-sulfur bacterium was isolated in pure culture from seawater collected from the seashore of Visakhapatnam, on the east coast of India, in a medium that contained 2 % NaCl (w/v). Strain JA123(T) was Gram-negative and non-motile and had a requirement for NaCl. Photo-organoheterotrophic and chemo-organoheterotrophic growth occurred with organic compounds as carbon sources and electron donors. Photolithoautotrophic, chemolithoautotrophic and fermentative growth could not be demonstrated. Strain JA123(T) contained vesicular intracellular photosynthetic membrane structures. Bacteriochlorophyll a and probably carotenoids of the spheroidene series were present as photosynthetic pigments. Biotin was required for growth. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA123(T) clustered with species of the genus Rhodobacter. Based on 16S rRNA gene sequence analysis and morphological and physiological characteristics, strain JA123(T) is sufficiently different from other Rhodobacter species to propose a novel species, Rhodobacter vinaykumarii sp. nov., to accommodate this strain; the type strain is JA123(T) (=DSM 18714(T) =JCM 14544(T) =CCUG 54311(T)).


Ghrelin plays a minor role in the physiological control of cardiac function in the rat.
We have previously reported that a 7-d pretreatment with hexarelin, a synthetic ligand of the GH secretagogue receptor (GHS-R), largely prevented damages induced by ischemia and reperfusion in isolated rat hearts. Our aim was to ascertain whether ghrelin, an endogenous ligand of the GHS-R, is physiologically endowed with cardioprotective activity. Hypophysectomized rats were treated in vivo for 7 d with either ghrelin (320 microg/kg) or hexarelin (80 microg/kg), and their hearts were subjected in vitro to the ischemia and reperfusion procedure. Ghrelin was far less effective than hexarelin in preventing increases in left ventricular end-diastolic pressure (15% and 60% protection for ghrelin and hexarelin, respectively), coronary perfusion pressure (10% and 45% reduction), and release of creatine kinase in the heart perfusate (15% and 55% reduction). In the second experiment, normal rats were passively immunized against ghrelin for 21 d before the ischemia and reperfusion procedure. In these isolated hearts, the ischemia-reperfusion damage was not significantly increased compared with control rats. After hypophysectomy, CD36 mRNA levels significantly increased, whereas those of atrial natriuretic factor significantly decreased. We conclude that: 1) ghrelin plays a minor role in the control of heart function; and 2) hexarelin effects are mediated in part by the GHS-R and largely by interactions with the CD36.


Relieving distressed caregivers (ReDiCare study): study protocol of a randomized pragmatic trial.
Providing care for someone with a disease or chronic condition can have a negative psychological, physical, social, and economic impact upon informal caregivers. Despite the socio-economic relevance and more than three decades of caregiver intervention research only very few translational efforts of successful interventions are reported. Still less of these interventions have been implemented into routine services. The aim of the ReDiCare study (German acronym BerTA) is to evaluate the effectiveness of a stepped counselling approach for burdened caregivers delivered by care counsellors of two long-term care insurances and registered psychotherapists. A pragmatic randomised controlled trial with 572 caregivers of older adults (≥ 60 years) receiving benefits of one of the two participating long-term care insurances. Participants are assigned (t0) to either the ReDiCare intervention or a control group receiving routine care and counselling. Data are collected at baseline (-t1), 3-month (t1), 9-month (t2) and 15-month (t3). The 9-month post-intervention assessment (t2) is the primary endpoint to evaluate the results on the primary and secondary outcomes, measured by self-reported questionnaires. Depressive symptoms measured with the CES-D are the primary outcome. The main secondary outcomes are physical complaints, utilization of psychosocial resources, caregiver self-efficacy and burden, positive aspects of caregiving and perceived care quality. A process evaluation, including audio tapes, self-report questionnaires and documentation will be conducted to examine internal and external validity of the intervention. Data on direct and indirect costs are collected for the (health) economic evaluation, using a health care perspective and a societal perspective. While comparable previous caregiver interventions have been developed and evaluated for specific caregiver groups (e.g. dementia caregivers, stroke caregivers), the ReDiCare study will indicate whether a stepped approach will be effective also in a broader group of caregivers. The intervention is one of the very few translational studies in caregiver intervention research and will provide valuable insights into relevant factors for training, intervention protocol adherence, effectiveness, and costs for future implementation steps. Deutsches Register Klinischer Studien (German Clinical Trials Register), DRKS00014593 ( www.drks.de, registered 14 May 2018) and International Clinical Trials Registry Platform, DRKS00014593 ( https://apps.who.int/trialsearch/ ).


Delayed-type hypersensitivity activity of the Brucella L7/L12 ribosomal protein depends on posttranslational modification.
The ribosomal protein L7/L12 isolated from Brucella melitensis induces a delayed-type hypersensitivity (DTH) reaction in brucella-sensitized guinea pigs. Surprisingly, the recombinant brucella L7/L12 protein expressed in Escherichia coli as a fusion protein with a six-histidine tag cannot elicit such a reaction. The six histidines tagged to the recombinant L7/L12 protein were removed enzymatically, but the resulting protein did not induce a DTH reaction in sensitized animals. Incubation of the recombinant L7/L12 fusion protein in a B. melitensis lysate endowed the recombinant protein with a DTH activity, suggesting that the recombinant protein was modified by this treatment. Glycosylation or phosphorylation of the recombinant L7/L12 protein could not be detected. On the other hand, radiolabeled palmitic acid was found to be incorporated to the recombinant protein during its incubation in the brucella lysate. This incorporation was specific for the brucella L7/L12 protein and was inhibited when the brucella lysate was frozen and thawed before the incubation. The data reported here indicate that posttranslational modification of L7/L12 protein comprising at least an acylation step is required for the brucella L7/L12 DTH activity.


Loqs depends on R2D2 to localize in D2 body-like granules and functions in RNAi pathways in silkworm cells.
The phenomenon of RNA interference (RNAi) has been found in various organisms. However, the proteins implicated in RNAi pathway in different species show distinct roles. Knowledge on the underlying mechanism of lepidopteron RNAi is quite lacking such as the roles of Loquacious (Loqs) and R2D2, the dsRNA-binding proteins in silkworm RNAi pathway. Here, we report that Loqs and R2D2 protein depletion affected efficiency of dsRNA-mediated RNAi pathway. Besides, Loqs was found to co-localize with Dicer2 to some specific cytoplasmic foci, which were looked like D2-bodies marked by R2D2 and Dicer2 in Fly cells, thereby calling the foci as D2 body-like granules. Using RNAi methods, Loqs was found to be the key protein in these granules, although R2D2 determined the localization of Loqs in D2 body-like granules. Interestingly, in the R2D2-depeted silkworm cells, the formation of processing bodies, another cytoplasmic foci, was affected. These data indicated R2D2 regulated these two kinds of cytoplasmic foci. Domain deletion analysis demonstrated that dsRBD 1 and 2 were required for Loqs in D2 body-like granules and dsRBD 2 and 3 were required for Loqs to interact with R2D2 and Ago1, respectively. Altogether, our observations provide important information for further study on D2 body-like granules, the newly found cytoplasmic foci in silkworm cells.