Transforming Healthcare Delivery: Integrating Dynamic Simulation Modelling and Big Data in Health Economics and Outcomes Research.
In the era of the Information Age and personalized medicine, healthcare delivery systems need to be efficient and patient-centred. The health system must be responsive to individual patient choices and preferences about their care, while considering the system consequences. While dynamic simulation modelling (DSM) and big data share characteristics, they present distinct and complementary value in healthcare. Big data and DSM are synergistic-big data offer support to enhance the application of dynamic models, but DSM also can greatly enhance the value conferred by big data. Big data can inform patient-centred care with its high velocity, volume, and variety (the three Vs) over traditional data analytics; however, big data are not sufficient to extract meaningful insights to inform approaches to improve healthcare delivery. DSM can serve as a natural bridge between the wealth of evidence offered by big data and informed decision making as a means of faster, deeper, more consistent learning from that evidence. We discuss the synergies between big data and DSM, practical considerations and challenges, and how integrating big data and DSM can be useful to decision makers to address complex, systemic health economics and outcomes questions and to transform healthcare delivery.


Social network analysis of Iranian researchers on emergency medicine: a sociogram analysis.
The purpose of this study was to report interaction patterns among Iranian authors of emergency medicine using social network analysis methodology, focusing on coauthorship network. The bibliographic data of Iranian authors on the 'emergency medicine' field during the years 2001-2011 were retrieved from the Science Citation Index Expanded database. Co-occurrence matrices were made by BibExcel and were imported to Ucinet and NetDraw to delineate coauthorship network. To detect structural patterns among authors, we considered some measures of social network analysis, such as density, centralisation indices, component analysis and cut-points. Lastly, subject experts separately analysed the content of papers. Of 116 papers published, the network was composed of 10 components, with the largest component having 25 authors. Using social network analysis measures, we identified science bottlenecks in knowledge sharing, hub authors and accelerators of information flow. Topic analysis showed 'Wounds and Injuries' as the most recent theme in all components because of existence of national registry for trauma, high burden of road traffic injuries and research priority of injuries in Iran. because of Iranian low productivity in the emergency medicine field, social network analysis seems to be a proper option for bibliometrics to identify central authors and detect knowledge structure in this field.


Functional overlap of the dictyostelium RasG, RasD and RasB proteins.
Disruption of the rasG gene in Dictyostelium discoideum results in several distinct phenotypes: a defect in cytokinesis, reduced motility and reduced growth. Reintroduction of the rasG gene restores all of the properties of the rasG(-) cells to those of the wild type. To determine whether the defects are due to impaired interactions with a single or multiple downstream effectors, we tested the ability of the highly related but non identical Dictyostelium ras genes, rasD and rasB, to rescue the defects. Introduction of the rasD gene under the control of the rasG promoter into rasG null (rasG(-)) cells corrected all phenotypes except the motility defect, suggesting that motility is regulated by a RasG mediated pathway that is different to those regulating growth or cytokinesis. Western blot analysis of RasD protein levels revealed that vegetative rasG(- )cells contained considerably more protein than the parental AX-3 cells, suggesting that RasD protein levels are negatively regulated in vegetative cells by RasG. The level of RasD was enhanced when the rasD gene was introduced under the control of the rasG promoter, and this increase in protein is presumably responsible for the reversal of the growth and cytokinesis defects of the rasG(- )cells. Thus, RasD protein levels are controlled by the level of RasG, but not by the level of RasD. Introduction of the rasB gene under the control of the rasG promoter into rasG(-) cells produced a complex phenotype. The transformants were extremely small and mononucleate and exhibited enhanced motility. However, the growth of these cells was considerably slower than the growth of the rasG(-) cells, suggesting the possibility that high levels of RasB inhibit an essential process. This was confirmed by expressing rasB in wild-type cells; the resulting transformants exhibited severely impaired growth. When RasB protein levels were determined by western blot analysis, it was found that levels were higher in the rasG(- )cells than they were in the wild-type parental, suggesting that RasG also negatively regulates rasB expression in vegetative cells. Overexpression of rasB in the rasG(- )cells also reduced the level of RasD protein. In view of the fact that alternate Ras proteins correct some, but not all, of the defects exhibited by the rasG(-) cells, we propose that RasG interacts with more than one downstream effector. In addition, it is clear that the levels of the various Ras proteins are tightly regulated in vegetative cells and that overexpression can be deleterious.


The development and use of mass media interventions for health-care messages about back pain: what do members of the public think?
The study sought to identify and explore the opinions, beliefs and views of members of the public regarding the use of media interventions for the delivery of health-care messages. It used a draft back pain campaign looking at the level of credibility, acceptability and trust in the authority of these messages. Sixty-eight members of the public participated in the study. A qualitative study design was used with semi-structured focus groups and a sampling frame including gender, age, socio-economic group, and experience of back pain. Three main themes were identified. 1. Media consumption. 2. Credibility. 3. Specific issues surrounding the proposed sample media campaign. The use of media to provide health-care information was viewed positively, with the NHS perceived as the most trustworthy source, and Government bodies viewed with scepticism. Issues surrounding the language, terminology and tone of campaigns were raised. A closer collaboration between health-care professionals and the public is advocated to achieve valuable and effective media campaigns.


Macrophages expressing triggering receptor expressed on myeloid cells-1 are underrepresented in the human intestine.
Triggering receptor expressed on myeloid cells (TREM)-1 is a cell surface molecule on neutrophils and monocytes/macrophages implicated in the amplification of inflammatory responses by enhancing degranulation and secretion of proinflammatory mediators. Macrophages play an important role in the intestinal mucosal immune system, because they are preferentially localized in the subepithelial region. Despite the presence of enormous numbers of bacteria in the colonic mucosa and the close proximity between mucosal macrophages and luminal bacteria, the intestinal mucosa normally displays minimal signs of inflammation. In this study, we show that the resident macrophage population in normal human small and large intestine contains only few TREM-1-expressing macrophages (<10%), whereas the overwhelming majority of monocytes (>90%) and macrophages from lymph nodes or tonsils (>80%) express TREM-1 on the cell surface. These findings were confirmed by FACS analysis and immunostainings of frozen tissue sections. The differential expression of TREM-1 greatly affects the functional capacities of monocytes and tissue macrophages. Although monocytes and macrophages from spleen, lymph nodes, or tonsils show a substantial increase in oxidative burst after TREM-1 cross-linking, no effect is seen in intestinal macrophages. Intriguingly, in contrast to monocytes, intestinal macrophages fail to up-regulate TREM-1 in response to TNF. This refractory state may be induced in intestinal macrophages by the local presence of IL-10 and TGF-beta, because these two immunoregulatory cytokines synergistically down-regulate TREM-1 expression on monocytes in vitro. The absence of TREM-1 expression on lamina propria macrophages is likely to prevent excessive inflammatory reactions, and thus, excessive tissue damage in the intestine.


All sealed endoleaks are not the same: a treatment strategy based on an ex-vivo analysis.
factors contributing to pressure transmission through thrombosed or sealed endoleaks have not been elucidated. The purpose of this investigation was to create an ex-vivo model that mimics patent and sealed endoleaks and that can quantitatively analyse the effects of length, diameter and thrombus on pressure transmission to the interior of the aneurysm sac. In the ex-vivo model, endoleak channels (ELCs) of various lengths (2 cm, 6 cm, 10 cm) and diameters (0.6 cm, 1.0 cm, 1.4 cm) were constructed using polytetrafluoroethylene (PTFE) grafts and attached to an artificial aneurysm sac. These ELCs were incorporated within a mock circulation made of rubber tubing connected to a pulsatile pump. Peak systolic pressure (PSP) was recorded in the aneurysm sac, distal to each ELC. Subsequently the ELCs were filled with human thrombus, and the pressure measurements repeated (n =5). Data was evaluated by regression analysis. Peak systolic pressure in the artificial circulation was maintained at 150 mmHg. In the absence of thrombus pressure did not change across the ELC, regardless of its length or diameter. In the presence of organised thrombus, the pressure curves distal to the ELC were dampened, and the pressure reduction was directly proportional to the length and inversely proportional to the diameter of the ELC. Regression analysis indicated statistical significance. In the absence of thrombosis, pressure transmitted via an ELC to the aneurysm sac is unchanged regardless of its length or diameter. All sealed endoleaks also transmit pressure. However, when an endoleak has thrombosed, pressure reduction is directly proportional to the length and inversely proportional to the diameter of its channel. This ex-vivo model suggests that Type 2 endoleaks with longer channels and smaller diameters would derive a greater benefit from adjunctive manoeuvres (coil embolisation) that hasten thrombosis. On the other hand, thrombosis of endoleaks with short and wide channels (e.g. Type 1) may not result in substantial pressure reduction within the aneurysm sac and a successful outcome.


National Readmission Patterns of Isolated Splenic Injuries Based on Initial Management Strategy.
Options for managing splenic injuries have evolved with a focus on nonoperative management. Long-term outcomes, such as readmissions and delayed splenectomy rate, are not well understood. To describe the natural history of isolated splenic injuries in the United States and determine whether patterns of readmission were influenced by management strategy. The Healthcare Cost and Utilization Project's Nationwide Readmission Database is an all-payer, all-ages, longitudinal administrative database that provides data on more than 35 million weighted US discharges yearly. The database was used to identify patients with isolated splenic injuries and the procedures that they received. Adult patients with isolated splenic injuries admitted from January 1 through June 30, 2013, and from January 1 through June 30, 2014, were included. Those who died during the index hospitalization or who had an additional nonsplenic injury with an Abbreviated Injury Score of 2 or greater were excluded. Univariate and mixed-effects logistic regression analysis controlling for center effect were used. Weighted numbers are reported. Initial management strategy at the time of index hospitalization, including nonprocedural management, angioembolization, and splenectomy. All-cause 6-month readmission rate. Secondary outcome was delayed splenectomy rate. A weighted sample of 3792 patients (2146 men [56.6%] and 1646 women [43.4%]; mean [SE] age, 48.5 [0.7] years) with 5155 admission events was included. During the index hospitalization, 825 (21.8%) underwent splenectomy, 293 (7.7%) underwent angioembolization, and 2673 (70.5%) had no procedure. The overall readmission rate was 21.1% (799 patients). Readmission rates did not differ based on initial management strategy (195 patients undergoing splenectomy [23.6%], 70 undergoing angioembolism [23.9%], and 534 undergoing no procedure [20%]; P = .33). Splenectomy was performed in 36 of 799 readmitted patients (4.5%) who did not have a splenectomy at their index hospitalization, leading to an overall delayed splenectomy rate of 1.2% (36 of 2967 patients). In mixed-effects logistic regression analysis controlling for patient, injury, clinical, and hospital characteristics, the choice of splenectomy (odds ratio, 0.93; 95% CI, 0.66-1.31) vs angioembolization (odds ratio, 1.19; 95% CI, 0.72-1.97) as initial management strategy was not associated with readmission. This national evaluation of the natural history of isolated splenic injuries from index admission through 6 months found that approximately 1 in 5 patients are readmitted within 6 months of discharge after an isolated splenic injury. However, the chance of readmission for splenectomy after initial nonoperative management was 1.2%. This finding suggests that the current management strategies used for isolated splenic injuries in the United States are well matched to patient need.


B7-H3 and its relevance in cancer; immunological and non-immunological perspectives.
B7-H3 is a transmembrane glycoprotein and a member of the B7 family of proteins. It was previously known as an immunoregulatory molecule, shown in recent years to be of clinical significance in different types of cancer. In some tumor types high expression of B7-H3 has been linked to a poor prognosis, whereas in other cancers the opposite effect has been observed. Taken together, the precise role of B7-H3 in tumor immunity is unclear and further investigations are needed. Another aspect of B7-H3 that so far has received little interest is its role in non-immunological systems. We have demonstrated that knockdown of B7-H3 in melanoma and breast cancer cells results in both increased chemosensitivity and decreased metastatic potential. This has been observed in both in vitro and in vivo experiments. Several different signaling pathways seems to be involved, as B7-H3 knockdown can be linked to both higher expression of apoptotic markers and increased phosphorylation of Stat3. Increased knowledge of also the non-immunological role of B7-H3 protein is therefore of great biological and putative therapeutic interest.


Pathophysiology of irreversible ischemic injury. The border zone controversy.
At the present time our experimental findings plus the weight of other experimental evidence suggest that there is unlikely to be a quantitatively significant border zone in the lateral plane. The transition from normal to ischemic tissue is likely to be accomplished over a distance of 1.0 mm or less and possibly in as little as the dimensions of one cell. The situation in the transmural plane is less well established, but if the same situation occurs, then the absence of a spatially indentifiable border zone of intermediate injury will require a major reappraisal, although not an abandonment, of concepts for the therapeutic limitation of infarct size. Any extrapolation of the observations, comments, and conclusions made in this paper to the human heart should be made with extreme caution. Major species differences exist, particularly in relation to the characteristics of collateral flow. Most experimental studies have involved single or multiple coronary artery ligation, a situation that generates large areas of sharply demarcated ischemia. These areas are very severely ischemic and short of reperfusion, which is hardly a practical consideration in the early phases of evolving myocardial infarction; the affected tissue is inevitably condemned to cell death and necrosis. The situation prevailing in man with partial coronary artery occlusion or diffuse ischemic heart disease may well be be very different and is clearly in urgent need of investigation.


Chemokine receptor 5 polymorphism in myocardial infarction patients from Luxembourg.
The CC-chemokine receptor 5 (CCR5) is regulating inflammatory pathways and may thus be implicated in the development and progression of heart failure (HF). A 32 base pair deletion of the ccr5 gene, called CCR5delta32, prevents the expression of CCR5 at the cell surface. We analyzed the association between the CCR5delta32 deletion and the risk and severity of myocardial infarction (MI) in a cohort of patients from Luxembourg. Using TaqMan allelic discrimination assay, we genotyped a total of 1080 patients undergoing coronary angiography. This population contained 3 groups of patients: controls with atypical chest pain, abnormal stress testing but normal coronary angiography (n = 154), patients with angina who underwent uncomplicated primary coronary intervention (n = 230), and patients with acute MI (n = 696). In MI patients, left ventricular ejection fraction (LVEF) was determined 1-month after MI with echocardiography. The frequency of the CCR5delta32 deletion was 16.3% in the global population, and was similar between controls, patients with angina and MI patients. The deletion was not associated with variations of plasma levels of creatine phosphokinase and troponin T, nor it was associated with LVEF, New York Heart Association class or 2-year mortality. The frequency of the deletion was comparable between MI patients with LV dysfunction (EF < or = 40%, n = 82) and no LV dysfunction (EF > 40%, n = 402). The frequency of the CCR5delta32 deletion in Luxembourg is similar to that observed in other European countries and is not associated with the risk of developing MI and LV dysfunction.


Course and predictors of rehospitalization in adolescent anorexia nervosa in a multisite study.
The course and clinical predictors of rehospitalization were studied in a large sample of adolescent patients with anorexia nervosa who had been treated at five European sites and been followed-up. Two-hundred and twelve adolescent patients first admitted for in-patient treatment, aged 10-18 (Mean 14.9) years and 94.8% female, were followed-up for an average of 8.3 years after first admission at sites in (former East and West Berlin, Zurich, Sofia, and Bucharest). Clinical history data were collected by use of standardized item-sheets at first admission. Semi-structured interviews including ratings of eating pathology and psychosocial outcome were performed at follow-up. About 44.8% (95/212) of the patients required at least one readmission. Rates of rehospitalization were significantly higher at the Eastern sites (Sofia and Bucharest). Significant predictors of readmission were paternal alcoholism, history of anorexia nervosa in the family, eating disorder in infancy, periodic overactivity, lower weight increase at first admission, and lower BMI at first discharge. In a model of logistic regression analysis, five variables (paternal alcoholism, eating disorder in infancy, periodic overactivity, low weight increase during first admission, and low BMI at first discharge) correctly classified 69% of the participants into cases with single or repeated admissions. Patients with repeated admissions had a less favorable long term outcome and had higher rates of persisting psychopathology at follow-up. Rehospitalization reflects the chronic course in a sizable proportion of adolescent patients with anorexia nervosa. Family psychopathology may have an effect both in terms of genetic vulnerability and environmental stress leading to unfavorable courses. The strong effect of insufficient weight gain during first admission and lower BMI at first discharge emphasizes the importance of adequate interventions. Readmissions carry the risk for later poor psychosocial and psychiatric outcomes.


The BRAzil MAGnesium (BRAMAG) trial: a randomized clinical trial of oral magnesium supplementation in pregnancy for the prevention of preterm birth and perinatal and maternal morbidity.
Preterm birth is the leading cause of infant mortality globally, including Brazil. We will evaluate whether oral magnesium citrate reduces the risk of placental dysfunction and its negative consequences for both the fetus and mother, which, in turn, should reduce the need for indicated preterm delivery. We will complete a multicenter, randomized double-blind clinical trial comparing oral magnesium citrate 150 mg twice daily (n = 2000 women) to matched placebo (n = 1000 women), starting at 121/7 to 206/7 weeks gestation and continued until delivery. We will include women at higher risk for placental dysfunction, based on clinical factors from a prior pregnancy (e.g., prior preterm delivery, stillbirth or preeclampsia) or the current pregnancy (e.g., chronic hypertension, pre-pregnancy diabetes mellitus, maternal age > 35 years or pre-pregnancy maternal body mass index > 30 kg/m2). The primary perinatal outcome is a composite of preterm birth < 37 weeks gestation, stillbirth > 20 weeks gestation, neonatal death < 28 days, or SGA birthweight < 3rd percentile. The primary composite maternal outcome is preeclampsia arising < 37 weeks gestation, severe non-proteinuric hypertension arising < 37 weeks gestation, placental abruption, maternal stroke during pregnancy or ≤ 7 days after delivery, or maternal death during pregnancy or ≤ 7 days after delivery. The results of this randomized clinical trial may be especially relevant in low and middle income countries that have high rates of prematurity and limited resources for acute newborn and maternal care. ClinicalTrials.gov Identifier NCT02032186, registered December 19, 2013.


The effect of maternal immunity on the equine gammaherpesvirus type 2 and 5 viral load and antibody response.
Two types of gammaherpesviruses (γEHV) are known to infect horses, EHV-2 and EHV-5. Foals become infected early in life, probably via the upper respiratory tract, despite maternal antibodies. In this study, we analyzed samples from a herd of mares and their foals. The foals were followed from birth to 22 months of age and the dams during the first 6 months postpartum. Blood and nasal swab samples were taken regularly for evaluation of antibody responses, virus isolation and viral load by qPCR. EHV-2 was isolated on day 5, and EHV-5 on day 12, earlier than previously reported. γEHV specific antibodies were not detectable in serum of foals before colostrum intake but peaked a few days after colostrum. Overall, EHV-2 viral load peaked in nasal swab at three to four months of age, paralleled with decline in maternal antibodies, but EHV-5 viral load did not peak until month 12. Maternal antibodies had a notable effect on the viral load and induction of endogenous antibody production. Foals were grouped in two groups depending on the mare's γEHV specific total IgG levels in serum at birth, group-high and group-low. Group-high had higher levels of maternal γEHV specific total IgG and IgG4/7 for the first 3 months, but when the endogenous production had superseded maternal antibodies, group-low was higher. The maternal antibodies had an effect on the γEHV viral load. Group-low peaked in EHV-2 viral load one month earlier than group-high. These effects were more evident for EHV-5, as there were seven months between the viral load peaks for the groups. The study provides information on how maternal antibody transfer affects γEHV shedding and antibody production in offspring. It also extends our knowledge on the occurrence of EHV-2 and EHV-5 infection in foals during the first two years of life.


Degronomics: Mapping the Interacting Peptidome of a Ubiquitin Ligase Using an Integrative Mass Spectrometry Strategy.
Human cells make use of hundreds of unique ubiquitin E3 ligases to ensure proteome fidelity and control cellular functions by promoting protein degradation. These processes require exquisite selectivity, but the individual roles of most E3s remain poorly characterized in part due to the challenges associated with identifying, quantifying, and validating substrates for each E3. We report an integrative mass spectrometry (MS) strategy for characterizing protein fragments that interact with KLHDC2, a human E3 that recognizes the extreme C-terminus of substrates. Using a combination of native MS, native top-down MS, MS of destabilized samples, and liquid chromatography MS, we identified and quantified a near complete fraction of the KLHDC2-binding peptidome in E. coli cells. This degronome includes peptides that originate from a variety of proteins. Although all identified protein fragments are terminated by diglycine or glycylalanine, the preceding amino acids are diverse. These results significantly expand our understanding of the sequences that can be recognized by KLHDC2, which provides insight into the potential substrates of this E3 in humans. We anticipate that this integrative MS strategy could be leveraged more broadly to characterize the degronomes of other E3 ligase substrate receptors, including those that adhere to the more common N-end rule for substrate recognition. Therefore, this work advances "degronomics," i.e., identifying, quantifying, and validating functional E3:peptide interactions in order to determine the individual roles of each E3.


Organism-specific depletion of highly abundant RNA species from bacterial total RNA.
High-throughput sequencing has become a standard tool for transcriptome analysis. The depletion of overrepresented RNA species from sequencing libraries plays a key role in establishing potent and cost-efficient RNA-seq routines. Commercially available kits are known to obtain good results for the reduction of ribosomal RNA (rRNA). However, we found that the transfer-messenger RNA (tmRNA) was frequently highly abundant in rRNA-depleted samples of Pseudomonas aeruginosa , consuming up to 25 % of the obtained reads. The tmRNA fraction was particularly high in samples taken from stationary cultures. This suggests that overrepresentation of this RNA species reduces the mRNA fraction when cells are grown under challenging conditions. Here, we present an RNase-H-based depletion protocol that targets the tmRNA in addition to ribosomal RNAs. We were able to increase the mRNA fraction to 93-99% and therefore outperform not only the commercially Ribo-off kit (Vazyme) operating by the same principle but also the formerly widely used Ribo-Zero kit (Illumina). Maximizing the read share of scientifically interesting RNA species enhances the discriminatory potential of next-generation RNA-seq experiments and, therefore, can contribute to a better understanding of the transcriptomic landscape of bacterial pathogens and their used mechanisms in host infection.


Classification of lattice defects in the kesterite Cu2ZnSnS4 and Cu2ZnSnSe4 earth-abundant solar cell absorbers.
The kesterite-structured semiconductors Cu2ZnSnS4 and Cu2ZnSnSe4 are drawing considerable attention recently as the active layers in earth-abundant low-cost thin-film solar cells. The additional number of elements in these quaternary compounds, relative to binary and ternary semiconductors, results in increased flexibility in the material properties. Conversely, a large variety of intrinsic lattice defects can also be formed, which have important influence on their optical and electrical properties, and hence their photovoltaic performance. Experimental identification of these defects is currently limited due to poor sample quality. Here recent theoretical research on defect formation and ionization in kesterite materials is reviewed based on new systematic calculations, and compared with the better studied chalcopyrite materials CuGaSe2 and CuInSe2 . Four features are revealed and highlighted: (i) the strong phase-competition between the kesterites and the coexisting secondary compounds; (ii) the intrinsic p-type conductivity determined by the high population of acceptor CuZn antisites and Cu vacancies, and their dependence on the Cu/(Zn+Sn) and Zn/Sn ratio; (iii) the role of charge-compensated defect clusters such as [2CuZn +SnZn ], [VCu +ZnCu ] and [ZnSn +2ZnCu ] and their contribution to non-stoichiometry; (iv) the electron-trapping effect of the abundant [2CuZn +SnZn ] clusters, especially in Cu2ZnSnS4. The calculated properties explain the experimental observation that Cu poor and Zn rich conditions (Cu/(Zn+Sn) ≈ 0.8 and Zn/Sn ≈ 1.2) result in the highest solar cell efficiency, as well as suggesting an efficiency limitation in Cu2ZnSn(S,Se)4 cells when the S composition is high.


Deep learning-based three-dimensional segmentation of the prostate on computed tomography images.
Segmentation of the prostate in computed tomography (CT) is used for planning and guidance of prostate treatment procedures. However, due to the low soft-tissue contrast of the images, manual delineation of the prostate on CT is a time-consuming task with high interobserver variability. We developed an automatic, three-dimensional (3-D) prostate segmentation algorithm based on a customized U-Net architecture. Our dataset contained 92 3-D abdominal CT scans from 92 patients, of which 69 images were used for training and validation and the remaining for testing the convolutional neural network model. Compared to manual segmentation by an expert radiologist, our method achieved 83 % ± 6 % for Dice similarity coefficient (DSC), 2.3 ± 0.6 mm for mean absolute distance (MAD), and 1.9 ± 4.0 cm 3 for signed volume difference ( Δ V ). The average recorded interexpert difference measured on the same test dataset was 92% (DSC), 1.1 mm (MAD), and 2.1 cm 3 ( Δ V ). The proposed algorithm is fast, accurate, and robust for 3-D segmentation of the prostate on CT images.


[CANCER PATIENTS FOLLOW-UP – CROATIAN SOCIETY FOR MEDICAL ONCOLOGY CLINICAL GUIDELINES Part V: melanoma, sarcomas, central nerve system tumors, lung cancer].
Treatment of oncological patients must be based upon multidisciplinary approach, and takes place in specialized oncological centers. By the end of a specific oncological treatment further follow-up is being managed mostly by the oncologists, but the role of the general practitioners becomes more important every day and therefore should be precisely defined. Nowadays, most of the existing follow-up guidelines are not being based on prospective studies, yet on the expert's opinion of a precise oncological center or specialists. The aim of the Croatian Society of Medical Oncology (CSMO) with these recommendations is to standardize and rationalize the diagnostic procedures' algorithm in follow–up of oncological patients after primary treatment, in patients with melanoma, sarcomas, central nerve system tumors and lung cancer.


Effects of 9 Months of Martial Arts Training on Cardiac Autonomic Modulation in Healthy Children and Adolescents.
The aim of the study was to evaluate the cardiac autonomic modulation after 9 months of martial arts practice in healthy children and adolescents. The study included 59 children and adolescents who were divided into 3 groups: judo, Muay Thai, and control. Heart rate variability was measured by a heart rate monitor, model Polar RS800CX. The intervention occurred twice a week on nonconsecutive days, lasting 60 minutes each session. A 1-way analysis of variance was used to compare participants at baseline. The comparisons between groups at baseline and after the intervention were carried out by a 2-way analysis of variance for repeated measures. After 9 months of intervention, significant increases were observed for root mean square successive differences, with higher values post compared with baseline (19.5%; P = .04). For SD1, an interaction effect was observed, with increased posttraining values compared with baseline (24.1%; P = .04) for the judo group. Qualitative analysis of the Poincaré plot showed greater dispersion of RR intervals, mainly beat to beat, after the judo intervention compared with the baseline. The Muay Thai and control groups presented no improvement. After 9 months of intervention, there were increases in cardiac autonomic modulation of children and adolescents participating in judo training. The practice of martial arts, such as judo, can be encouraged from an early age to improve cardiovascular system functioning, possibly providing protection against cardiovascular problems.


Radiologic outcome and its relationship to functional disability in juvenile rheumatoid arthritis.
To determine the radiologic outcome in juvenile rheumatoid arthritis (JRA) and the relationship of radiologically detected joint damage to functional disability using multivariate analyses. Selection criteria included a diagnosis of JRA made by 1977 American College of Rheumatology criteria, onset of arthritis > or = 5 years prior to study, current age > or = 8 years, a minimum grade 3 reading ability, and the availability of radiographs. Disability was measured by the Childhood Health Assessment Questionnaire (CHAQ) and Steinbrocker classifications. Radiographs taken within 2 years after onset (early) and the most recent radiographs (late) were examined by a single pediatric radiologist blinded to patients' identities, diagnoses, and outcomes. Multiple regression analyses were performed. On late radiographs the frequencies of joint space narrowing were 38, 14, 43, and 79%, respectively, among patients with systemic, pauciarticular, rheumatoid factor (RF) negative polyarticular, and RF positive polyarticular onset; erosions occurred in 63, 25, 39, and 75%, respectively. Early erosions were most frequent in patients with RF+ polyarticular onset, while both joint space narrowing and erosions occurred early in systemic onset. Radiologic signs of joint damage were most frequent at hips and wrists, while knees and ankles were relatively spared. Based on patients who had radiographs performed within one year of clinical study, 17.7% of the variation in CHAQ score was explained by joint space narrowing, 32.4% by pain, and 5% by a severe rating on physician's global estimate of disease activity. The odds of a Steinbrocker class > I were increased by joint space narrowing, pain, systemic onset, and active joint count. Differences in the frequencies and patterns of joint damage occur both among JRA onset subtypes and among individual joints. Radiographic damage, especially joint space narrowing, correlates with functional disability. However, pain is the major contributor to variation in CHAQ scores.


Controlled single-cell deposition and patterning by highly flexible hollow cantilevers.
Single-cell patterning represents a key approach to decouple and better understand the role and mechanisms of individual cells of a given population. In particular, the bottom-up approach of engineering neuronal circuits with a controlled topology holds immense promises to perceive the relationships between connectivity and function. In order to accommodate these efforts, highly flexible SU-8 cantilevers with integrated microchannels have been fabricated for both additive and subtractive patterning. By directly squeezing out single cells onto adhesive surfaces, controlled deposition with a spatial accuracy of 5 μm could be achieved, while subtractive patterning has been realized by selective removal of targeted single cells. Complex cell patterns were created on substrates pre-patterned with cell-adhesive and repulsive areas, preserving the original pattern geometry for long-term studies. For example, a circular loop with a diameter of 530 μm has been realized using primary hippocampal neurons, which were fully connected to their respective neighbors along the loop. Using the same cantilevers, the versatility of the technique has also been demonstrated via in situ modification of already mature neuronal cultures by both detaching individual cells of the population and adding fresh ones, incorporating them into the culture.


Drug utilisation among older inpatients in a teaching hospital in Western Nepal.
The proportion of the elderly population in Nepal, though low, is steadily increasing. Studies on drug utilisation among geriatric patients in Western Nepal, a region with the highest proportion of the elderly in all of Nepal, are lacking. The present study was carried out at the Manipal Teaching Hospital, a 700-bed teaching hospital in Pokhara, Western Nepal. The study was carried out from June 1, 2005 to May 31, 2006 using case records of discharged patients aged 60 years and older. This was an observational study that utilised a retrospective case notes review methodology. The microorganisms isolated on culture and their antibiotic sensitivity patterns were studied. Drug consumption was measured by the defined daily dose (DDD) per 100 bed-days. The mean cost of the drugs was calculated. The percentage of drugs prescribed by generic name and the percentages of fixed-dose combinations used were noted. The basic demographic information of older inpatients, drug utilisation patterns, drug consumption using DDD, organisms isolated and their antibiotic sensitivity patterns, and the mean cost of the drugs were studied. A total of 548 patients were admitted. Hypertension, acute exacerbation of chronic obstructive pulmonary disease and cancer were the most common illnesses among the patients. The mean number of drugs consumed by each patient was 7.73. Escherichia coli and Pseudomonas aeruginosa were the most common microorganisms isolated. Ranitidine, multivitamins, amlodipine, ipratropium and dextrose normal saline were most commonly prescribed. The DDD per 100 bed-days of ranitidine and omeprazole (highest values) was 33.48 and 3.51, respectively. The mean cost of drugs was USD26.6, and antibiotics accounted for around 40 percent of the cost. The use of a high number of drugs, and of parenteral drugs and intravenous fluids, is a common problem. The prescription of generic drugs is low. Guidelines for the use of drugs in the elderly are required and further studies are needed on this issue.


A novel cry2Ab gene from the indigenous isolate Bacillus thuringiensis subsp. kurstaki.
A novel cry2Ab gene was cloned and sequenced from the indigenous isolate of Bacillus thuringiensis subsp. kurstaki. This gene was designated as cry2Ab25 and its sequence revealed an open reading frame of 1,902 bp encoding a 633 aa protein with calculated molecular mass of 70 kDa and pI value of 8.98. The amino acid sequence of the Cry2Ab25 protein was compared with previously known Cry2Ab toxins, and the phylogenetic relationships among them were determined. The deduced amino acid sequence of the Cry2Ab25 protein showed 99% homology to the known Cry2Ab proteins, except for Cry2Ab10 and Cry2Ab12 with 97% homology, and a variation in one amino acid residue in comparison with all known Cry2Ab proteins. The cry2Ab25 gene was expressed in Escherichia coli BL21(DE3) cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the Cry2Ab25 protein is about 70 kDa. The toxin expressed in BL21(DE3) exhibited high toxicity against Malacosoma neustria and Rhagoletis cerasi with 73% and 75% mortality after 5 days of treatment, respectively.


Developing novel treatments for mood disorders: accelerating discovery.
This review was generated from discussions by the Pharmacologic and Somatic Treatments Section of the National Institute of Mental Health Strategic Plan for Mood Disorders Committee on advancing novel pharmacologic and somatic treatments for mood disorders. The opening section of the article summarizes in broad strokes, current pharmacologic treatments, and new directions in the field. Thereafter the topics focus on specific research initiatives that could advance the current therapeutics for mood disorders including new basic and clinical research in vivo human imaging procedures, somatic therapeutics, and the vast new area of pharmacogenetics. New scientific and technical opportunities exist today based on advances in basic neuroscience, opportunities in clinical testing, industry interest in advancing central nervous system therapeutics, and on active consumer advocacy groups. The question of how to bring all of these positive forces together to accelerate discovery in mood disorder thera-peutics is the topic of this article.


New brain lesions in a patient with sarcoidosis: is it neurosarcoidosis?
A 45-year-old woman with pulmonary sarcoidosis diagnosed 5 years previously, who was on treatment with prednisone and methotrexate for 1year, developed partial seizure with secondary generalization. MRI showed three non-cavitary enhancing lesions in the cerebello-occipital region. These lesions were presumed to be neurosarcoidosis. Methotrexate was discontinued, prednisone dose was increased and azathiopurine and levetiracetam were added. While on treatment, follow up imaging showed enlarging brain lesions. Biopsy of the lesions showed Epstein Barr virus (EBV) positive diffuse B cell lymphoma. Immunosuppressants were tapered off and she was begun on Rituximab. Because of lack of improvement after 4 cycles of Rituximab, she was then treated with high dose Methotrexate and Temozolamide. We present this case as a diagnostic challenge. New enhancing brain lesions occurring in a patient with long standing sarcoidosis, while likely to be neurosarcoidosis, may be due to a complication of immunosuppressant therapy. The need for early biopsy, if the lesions do not improve, should be considered.


The Effect of Volitional Preemptive Abdominal Contraction on Biomechanical Measures During A Front Versus Back Loaded Barbell Squat.
Weightlifting is growing in popularity among recreational and competitive athletes. The barbell back squat (BackS) is commonly included in these training programs, while the barbell front squat (FrontS) is commonly performed as a component of other lifts such as the power clean or clean and jerk, it is less commonly practiced in isolation. The purpose of this study was to examine the effects of VPAC performance on trunk muscle and LE biomechanical responses during loaded BackS versus FrontS in healthy subjects. Controlled Laboratory Study. Healthy male subjects with the ability to perform a sub-maximal loaded barbell squat lift were recruited. Subjects completed informed consent, demographic/medical history questionnaires and an instructional video. Subjects practiced VPAC and received feedback. Surface electromyography (sEMG) electrodes and kinematic markers were applied. Muscles included were the internal oblique (IO), external oblique (EO), rectus abdominis, iliocostalis lumborum (ICL), superficial multifidi, rectus femoris, vastus lateralis, biceps femoris, and gluteus maximus. Maximal voluntary isometric contractions established reference sEMG values. A squat one-rep-max (1RM) was predicted by researchers using a three to five repetition maximum (3RM, 5RM) load protocol. Subjects performed BackS trials at 75% 1RM while FrontS trials were performed at 75% BackS weight, both with and without VPAC. Subjects performed three repetitions of each condition with feet positioned on two adjacent force plates. Significant interactions and main effects were tested using a 2(VPAC strategy) x 2(squat variation) and 2(VPAC strategy) x 2(direction) within-subject repeated measures ANOVAs. Tukey's Post-Hoc tests identified the location of significant differences. Trunk muscle activity was significantly higher during FrontS versus BackS regardless of VPAC condition. (IO: p=0.018, EO: p<0.001, ICL: p<0.001) VPAC increased performance time for both squat variations (p=.0011), which may be associated with decreased detrimental force potential on the lumbar spine and knees. VPAC led to improved ability to maintain a neutral lumbar spine during both squat variations. This finding is associated with decreased detrimental force potential on the lumbar spine. Findings could help guide practitioners and coaches to choose squat variations and incorporate VPAC strategies during their treatments and/or training programs. Level 3©The Author(s).


Teaching research in clinical pastoral education: a survey of model practices.
The Association of Professional Chaplains (APC) developed Standards of Practice for Acute and Long-term settings. Standard 12 promotes research-literate chaplains as important for the profession. Since many chaplains receive training in clinical pastoral education (CPE) residency programs, the aim of this study was to identify model practices for the teaching of research in such programs. Using a purposeful sample, this study identified 11 programs that offered "consistent and substantive" education in research. Common features included the existence of a research champion, a culture supportive of research, and the availability of institutional resources. The study identified models and methodologies that CPE programs can adopt.


Interaction of Alzheimer's beta -amyloid precursor family proteins with scaffold proteins of the JNK signaling cascade.
We have isolated a novel protein based on its association with Drosophila APP-like protein (APPL), a homolog of the beta-amyloid precursor protein (APP) that is implicated in Alzheimer's disease. This novel APPL-interacting protein 1 (APLIP1) contains a Src homology 3 domain and a phosphotyrosine interaction domain and is expressed abundantly in neural tissues. The phosphotyrosine interaction domain of APLIP1 interacts with a sequence containing GYENPTY in the cytoplasmic domain of APPL. APLIP1 is highly homologous to the carboxyl-terminal halves of mammalian c-Jun NH(2)-terminal kinase (JNK)-interacting protein 1b (JIP1b) and 2 (JIP2), which also contain Src homology 3 and phosphotyrosine interaction domains. The similarity of APLIP1 to JIP1b and JIP2 includes interaction with component(s) of the JNK signaling pathway and with the motor protein kinesin and the formation of homo-oligomers. JIP1b interacts strongly with the cytoplasmic domain of APP (APPcyt), as APLIP1 does with APPL, but the interaction of JIP2 with APPcyt is weak. Overexpression of JIP1b slightly enhances the JNK-dependent threonine phosphorylation of APP in cultured cells, but that of JIP2 suppresses it. These observations suggest that the interactions of APP family proteins with APLIP1, JIP1b, and JIP2 are conserved and play important roles in the metabolism and/or the function of APPs including the regulation of APP phosphorylation by JNK. Analysis of APP family proteins and their associated proteins is expected to contribute to understanding the molecular process of neural degeneration in Alzheimer's disease.


Biological effects of red beetroot and betalains: A review.
Over the past few years, the use of natural substances as protective or therapeutic agents has gained much attention worldwide. Recent modern studies have shown a variety of health benefits for red beetroot and its active compounds betalains (also betanin) such as antioxidative, anti-inflammation, anticancer, blood pressure and lipid lowering, also antidiabetic and anti-obesity effects. Betanin, the main component of the red beetroot, is a betalain glycosidic pigment, which is used as a food additive. This review summarizes findings in the literature and shows the therapeutic potential of red beetroot and its active compounds (betalains) as promising alternatives for supplemental therapies in multiple diseases.


Modulation of brain hemichannels and gap junction channels by pro-inflammatory agents and their possible role in neurodegeneration.
In normal brain, neurons, astrocytes, and oligodendrocytes, the most abundant and active cells express pannexins and connexins, protein subunits of two families forming membrane channels. Most available evidence indicates that in mammals endogenously expressed pannexins form only hemichannels and connexins form both gap junction channels and hemichannels. Whereas gap junction channels connect the cytoplasm of contacting cells and coordinate electric and metabolic activity, hemichannels communicate the intra- and extracellular compartments and serve as a diffusional pathway for ions and small molecules. A subthreshold stimulation by acute pathological threatening conditions (e.g., global ischemia subthreshold for cell death) enhances neuronal Cx36 and glial Cx43 hemichannel activity, favoring ATP release and generation of preconditioning. If the stimulus is sufficiently deleterious, microglia become overactivated and release bioactive molecules that increase the activity of hemichannels and reduce gap junctional communication in astroglial networks, depriving neurons of astrocytic protective functions, and further reducing neuronal viability. Continuous glial activation triggered by low levels of anomalous proteins expressed in several neurodegenerative diseases induce glial hemichannel and gap junction channel disorders similar to those of acute inflammatory responses triggered by ischemia or infectious diseases. These changes are likely to occur in diverse cell types of the CNS and contribute to neurodegeneration during inflammatory process.


Effect of varying water-to-powder ratios and ultrasonic placement on the compressive strength of mineral trioxide aggregate.
The purpose of this study was to compare the compressive strength of mineral trioxide aggregate (MTA) when mixed with 2 different water-to-powder (WP) proportions using either hand or ultrasonic placement. Tooth-colored ProRoot MTA (Dentsply Maillefer, Ballaigues, Switzerland) and white MTA Angelus (Angelus Soluçoes Odontologicas, Londrina, Brazil) were investigated. One gram of each MTA powder was mixed with either 0.34 or 0.40 g distilled water. The 4 groups were further divided into 2 groups of 5 specimens for each of the following techniques: conventional (ie, hand placement) and placement using indirect ultrasonic activation for 30 seconds. All specimens were subjected to compressive strength testing after 4 days. The results were statistically analyzed with multivariate analysis of variance and Tukey Honestly Significant Difference tests at a significance level of P < .05. The mean compressive strength values of ProRoot MTA (84.17 ± 22.68) were significantly greater than those of MTA Angelus (47.71 ± 14.29) (P < .01). Specimens mixed with the 0.34 WP ratio had higher compressive strength values (72.85 ± 25.77) than those mixed with the 0.40 WP ratio (56.69 ± 24.85) (P < .05). The highest compressive strength values were recorded for ProRoot MTA specimens that were mixed in the 0.34 WP ratio, and then the samples were placed with ultrasonic activation (mean = 91.35 MPa). The lowest values were recorded for MTA Angelus samples that were mixed in the 0.40 WP ratio, and the specimens were placed without ultrasonic activation (mean = 36.36 MPa). Ultrasonic activation had no significant difference in terms of compressive strength. When using ProRoot MTA and MTA Angelus, higher WP ratios resulted in lower compressive strength values. Ultrasonication had no significant effect on the compressive strength of the material regardless of the WP ratio that was used. Therefore, adherence to the manufacturer's recommended WP ratio when preparing MTA for use in dental applications is advised.


Experimentally Manipulated Low Social Status and Food Insecurity Alter Eating Behavior Among Adolescents: A Randomized Controlled Trial.
This randomized trial experimentally manipulated social status to assess effects on acute eating behavior and 24-hour energy balance. Participants (n = 133 Hispanics; age 15-21 years; 60.2% females) were randomized to low social status ("LOW") or high social status ("HIGH") conditions in a rigged game of Monopoly (Hasbro, Inc.). Acute energy intake in a lunchtime meal was measured by food scales. Twenty-four-hour energy balance was assessed via summation of resting metabolic rate (metabolic cart), physical activity energy expenditure (accelerometer), thermic effect of food, and subtraction of twenty-four-hour energy intake (food diary). In the total sample, no significant differences were observed by study condition at lunchtime. LOW females consumed a greater percent of lunchtime daily energy needs (37.5%) relative to HIGH females (34.3%); however, this difference was not statistically significant (P = 0.291). In males, however, LOW consumed significantly less (36.5%) of their daily energy needs relative to HIGH males (45.8%; P = 0.001). For 24-hour energy balance, sex differences were nearly significant (P = 0.057; LOW females: surplus +200 kcal; HIGH males: surplus +445 kcal). Food-insecure individuals consumed a nearly significant greater lunchtime percent daily energy than those with food security (40.7% vs. 36.3%; P = 0.0797). The data demonstrate differential acute and 24-hour eating behavior responses between Hispanic male and female adolescents in experimentally manipulated conditions of low social status.


The recombination signal sequence-binding protein RBP-2N functions as a transcriptional repressor.
We have identified a cellular protein, RBP-2N, a presumed recombinase, as a repressor of transcription. Inhibition of transcription by RBP-2N was dependent on its DNA recognition site and was demonstrated in vitro and in vivo. This repression appears to be general, as transcription mediated by SP1 and Gal4/VP16 was inhibited by RBP-2N. The protein was purified to near homogeneity from human cells on the basis of its binding to a site present in the promoter of the adenovirus pIX gene. The DNA recognition sequence is 5'-TGGGAAAGAA, which is markedly different from the recombination signal sequence originally identified as the target site for this protein. The sequence of the purified protein is 97% identical with that published for the mouse RBP-2N protein. The reported homolog in Drosophila is Suppressor of Hairless. RBP-2N binding sites are present in a number of cellular and viral promoters, so RBP-2N may have a general role in transcriptional repression.


Frequency-dependent optical pumping in atomic ?-systems.
We consider the effects of optical pumping on the conversion of laser-frequency modulation into intensity modulation by an atomic absorption line in a vapor of alkali atoms driven in a ?-configuration. It is found that, due to optical pumping in combination with the excited-state hyperfine structure, the absorption line shape is distorted substantially as the Fourier frequency of the FM is changed. The most significant effect of the distortion is a shift of the apparent line center, which depends on how the frequency of the modulation compares with the optical pumping rate. This shift has implications for locking lasers to atomic transitions and also for FM-AM noise conversion in atomic vapors.


Autophagy in protists.
Autophagy is the degradative process by which eukaryotic cells digest their own components using acid hydrolases within the lysosome. Originally thought to function almost exclusively in providing starving cells with nutrients taken from their own cellular constituents, autophagy is in fact involved in numerous cellular events including differentiation, turnover of macromolecules and organelles, and defense against parasitic invaders. During the last 10-20 years, molecular components of the autophagic machinery have been discovered, revealing a complex interactome of proteins and lipids, which, in a concerted way, induce membrane formation to engulf cellular material and target it for lysosomal degradation. Here, our emphasis is autophagy in protists. We discuss experimental and genomic data indicating that the canonical autophagy machinery characterized in animals and fungi appeared prior to the radiation of major eukaryotic lineages. Moreover, we describe how comparative bioinformatics revealed that this canonical machinery has been subject to moderation, outright loss or elaboration on multiple occasions in protist lineages, most probably as a consequence of diverse lifestyle adaptations. We also review experimental studies illustrating how several pathogenic protists either utilize autophagy mechanisms or manipulate host-cell autophagy in order to establish or maintain infection within a host. The essentiality of autophagy for the pathogenicity of many parasites, and the unique features of some of the autophagy-related proteins involved, suggest possible new targets for drug discovery. Further studies of the molecular details of autophagy in protists will undoubtedly enhance our understanding of the diversity and complexity of this cellular phenomenon and the opportunities it offers as a drug target.


An autonomously self-assembling dendritic DNA nanostructure for target DNA detection.
There is a growing need for sensitive and reliable nucleic acid detection methods that are convenient and inexpensive. Responsive and programmable DNA nanostructures have shown great promise as chemical detection systems. Here, we describe a DNA detection system employing the triggered self-assembly of a novel DNA dendritic nanostructure. The detection protocol is executed autonomously without external intervention. Detection begins when a specific, single-stranded target DNA strand (T) triggers a hybridization chain reaction (HCR) between two, distinct DNA hairpins (α and β). Each hairpin opens and hybridizes up to two copies of the other. In the absence of T, α and β are stable and remain in their poised, closed-hairpin form. In the presence of T, α hairpins are opened by toe-hold mediated strand-displacement, each of which then opens and hybridizes two β hairpins. Likewise, each opened β hairpin can open and hybridize two α hairpins. Hence, each layer of the growing dendritic nanostructure can in principle accommodate an exponentially increasing number of cognate molecules, generating a high molecular weight nanostructure. This HCR system has minimal sequence constraints, allowing reconfiguration for the detection of arbitrary target sequences. Here, we demonstrate detection of unique sequence identifiers of HIV and Chlamydia pathogens.


Patient-Reported Symptoms Improve Performance of Risk Prediction Models for Emergency Department Visits Among Patients With Cancer: A Population-Wide Study in Ontario Using Administrative Data.
Prior work shows measurements of symptom severity using the Edmonton Symptom Assessment System (ESAS) which are associated with emergency department (ED) visits in patients with cancer; however, it is not known if symptom severity improves the ability to predict ED visits. To determine whether information on symptom severity improves the ability to predict ED visits among patients with cancer. This was a population-based study of patients who were diagnosed with cancer and had at least one ESAS assessment completed between 2007 and 2015 in Ontario, Canada. After splitting the cohort into training and test sets, two ED visit risk prediction models using logistic regression were developed on the training cohort, one without ESAS and one with ESAS. The predictive performance of each risk model was assessed on the test cohort and compared with respect to area under the curve and calibration. The full cohort consisted of 212,615 unique patients with a total of 1,267,294 ESAS assessments. The risk prediction model including ESAS was superior in sensitivity, specificity, accuracy, and discrimination. The area under the curve was 73.7% under the model with ESAS, whereas it was 70.1% under the model without ESAS. The model with ESAS was also better calibrated. This improvement in calibration was particularly noticeable among patients in the higher deciles of predicted risk. This study demonstrates the importance of incorporating symptom measurements when developing an ED visit risk calculator for patients with cancer. Improved predictive models for ED visits using measurements of symptom severity may serve as an important clinical tool to prompt timely interventions by the cancer care team before an ED visit is necessary.


Large-scale identification of chemically induced mutations in Drosophila melanogaster.
Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-genome sequencing (WGS), it is now possible to sequence hundreds of strains, but determining which mutations are causative among thousands of polymorphisms remains challenging. We have sequenced 394 mutant strains, generated in a chemical mutagenesis screen, for essential genes on the Drosophila X chromosome and describe strategies to reduce the number of candidate mutations from an average of -3500 to 35 single-nucleotide variants per chromosome. By combining WGS with a rough mapping method based on large duplications, we were able to map 274 (-70%) mutations. We show that these mutations are causative, using small 80-kb duplications that rescue lethality. Hence, our findings demonstrate that combining rough mapping with WGS dramatically expands the toolkit necessary for assigning function to genes.


The evolution of the huntingtin-associated protein 40 (HAP40) in conjunction with huntingtin.
The huntingtin-associated protein 40 (HAP40) abundantly interacts with huntingtin (HTT), the protein that is altered in Huntington's disease (HD). Therefore, we analysed the evolution of HAP40 and its interaction with HTT. We found that in amniotes HAP40 is encoded by a single-exon gene, whereas in all other organisms it is expressed from multi-exon genes. HAP40 co-occurs with HTT in unikonts, including filastereans such as Capsaspora owczarzaki and the amoebozoan Dictyostelium discoideum, but both proteins are absent from fungi. Outside unikonts, a few species, such as the free-living amoeboflagellate Naegleria gruberi, contain putative HTT and HAP40 orthologs. Biochemically we show that the interaction between HTT and HAP40 extends to fish, and bioinformatic analyses provide evidence for evolutionary conservation of this interaction. The closest homologue of HAP40 in current protein databases is the family of soluble N-ethylmaleimide-sensitive factor attachment proteins (SNAPs). Our results indicate that the transition from a multi-exon to a single-exon gene appears to have taken place by retroposition during the divergence of amphibians and amniotes, followed by the loss of the parental multi-exon gene. Furthermore, it appears that the two proteins probably originated at the root of eukaryotes. Conservation of the interaction between HAP40 and HTT and their likely coevolution strongly indicate functional importance of this interaction.


31P magnetic resonance spectroscopy demonstrates expansion of the extracellular space in the skeletal muscle of starved rats.
Starvation significantly alters the distribution of body water. To study the effects of starvation on cellular energetics and water distribution in skeletal muscle, a novel 31P magnetic resonance technique (31P MRS) was developed to measure water compartments. After 31P MRS-visible water space markers which distribute in total body water (dimethyl methylphosphonate, DMMP) and extracellular water (phenylphosphonate, PPA) were infused intravenously, 31P MRS spectra were obtained from the gastrocnemius muscle of male virus-free Wistar rats at baseline and after starvation or ad libitum feeding for 4 days. Muscle water spaces were also measured using the chloride method and Nernst's equation. Muscle water contents as determined by drying were equivalent in the two groups. In vivo measurements of changes in DMMP relative to all of the MRS visible phosphates also demonstrated that the total water space was similar in control and starved rats. However, starvation significantly increased the ratio of PPA/DMMP (0.67 +/- 0.05 vs 0.87 +/- 0.04, Control vs Starvation; P < 0.001), and therefore the ratio of extracellular water to total water in the gastrocnemius. Furthermore, because muscle water contents were comparable between the groups, this expansion of the extracellular space was accompanied by contraction of the intracellular compartment in starved animals. Equivalent changes were detected in vitro using the chloride method. Lastly, phosphocreatine/ATP ratios, which measured changes in high-energy phosphate stores, decreased after starvation (4.09 +/- 0.06 vs 3.61 +/- 0.06; P < 0.001) and were inversely related to changes in PPA/DMMP (r = -0.61; P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


PDZ domain proteins: plug and play!
Protein-protein interactions are key elements in building functional protein complexes. Among the plethora of domains identified during the last 10 years, PDZ domains are one of the most commonly found protein-protein interaction domains in organisms from bacteria to humans. Although they may be the sole protein interaction domain within a cytoplasmic protein, they are most often found in combination with other protein interaction domains (for instance, SH3, PTB, WW) participating in complexes that facilitate signaling or determine the localization of receptors. Diversity of PDZ-containing protein function is provided by the large number of PDZ proteins that Mother Nature has distributed in the genome and implicates this protein family in the wiring of a huge number of molecules in molecular networks from the plasma membrane to the nucleus. Although at first sight their binding specificity appeared rather monotonous, involving only binding to the carboxyl-terminus of various proteins, it is now recognized that PDZ domains interact with greater versatility through PDZ-PDZ domain interaction; they bind to internal peptide sequences and even to lipids. Furthermore, PDZ domain-mediated interactions can sometimes be modulated in a dynamic way through target phosphorylation. In this review, we attempt to describe the structural basis of PDZ domain recognition and to give some functional insights into their role in the scaffolding of protein complexes implicated in normal and pathological biological processes.


In Silico Design and Experimental Validation of siRNAs Targeting Conserved Regions of Multiple Hepatitis C Virus Genotypes.
RNA interference (RNAi) is a post-transcriptional gene silencing mechanism that mediates the sequence-specific degradation of targeted RNA and thus provides a tremendous opportunity for development of oligonucleotide-based drugs. Here, we report on the design and validation of small interfering RNAs (siRNAs) targeting highly conserved regions of the hepatitis C virus (HCV) genome. To aim for therapeutic applications by optimizing the RNAi efficacy and reducing potential side effects, we considered different factors such as target RNA variations, thermodynamics and accessibility of the siRNA and target RNA, and off-target effects. This aim was achieved using an in silico design and selection protocol complemented by an automated MysiRNA-Designer pipeline. The protocol included the design and filtration of siRNAs targeting highly conserved and accessible regions within the HCV internal ribosome entry site, and adjacent core sequences of the viral genome with high-ranking efficacy scores. Off-target analysis excluded siRNAs with potential binding to human mRNAs. Under this strict selection process, two siRNAs (HCV353 and HCV258) were selected based on their predicted high specificity and potency. These siRNAs were tested for antiviral efficacy in HCV genotype 1 and 2 replicon cell lines. Both in silico-designed siRNAs efficiently inhibited HCV RNA replication, even at low concentrations and for short exposure times (24h); they also exceeded the antiviral potencies of reference siRNAs targeting HCV. Furthermore, HCV353 and HCV258 siRNAs also inhibited replication of patient-derived HCV genotype 4 isolates in infected Huh-7 cells. Prolonged treatment of HCV replicon cells with HCV353 did not result in the appearance of escape mutant viruses. Taken together, these results reveal the accuracy and strength of our integrated siRNA design and selection protocols. These protocols could be used to design highly potent and specific RNAi-based therapeutic oligonucleotide interventions.


Intrabiliary MR imaging: assessment of biliary obstruction with use of an intraluminal MR receiver coil.
The primary aim of this study was to determine whether intrabiliary magnetic resonance (MR) imaging is feasible in a clinical setting and to optimize MR imaging parameters for the technique. In addition, it was attempted to determine the accuracy of intrabiliary MR imaging in the setting of biliary obstruction of unknown cause. Intrabiliary MR was performed prospectively in 15 patients with biliary obstruction of unknown cause. A 0.030-inch MR intravascular receiver coil was placed in an existing biliary tube. Intrabiliary MR was performed on a 1.5-T system. T1-weighted, T2-weighted, and single-shot fast spin-echo images were acquired. T1-weighted images were also acquired after the administration of a gadolinium contrast agent. Signal intensity analysis was conducted in the region of the common bile duct. Accuracy of intrabiliary MR, computed tomography (CT), MR, and cholangiography were determined by correlation with surgical pathologic findings. Intrabiliary MR was successfully performed in 14 of 15 patients. MR examinations were performed in less than 1 hour. The signal-to-noise ratio in the region of the common bile duct with the intrabiliary MR technique was increased by a factor of 9 compared with standard surface-coil MR imaging (P < .00001). The mean n-plane resolution achieved was 740 +/- 20 microm x 1,150 +/- 20 microm obtained with use of a field of view of 18 cm x 18 cm (range, 15-24 cm) and a matrix of 256 x 160. Of the pulse sequences tested, the gadolinium-enhanced T1-weighted image was the best for identifying tumor and delineating tumor margins. Intrabiliary MR had a higher sensitivity than CT (100% vs 50%), a higher specificity than cholangiography (80% vs 20%), and a better correlation (P = .015) with surgical pathologic findings than CT, MR imaging, or cholangiography. Intrabiliary MR was well tolerated in a clinical setting and provided high spatial resolution and excellent contrast between the biliary lumen and adjacent structures. Intrabiliary MR demonstrated an advantage in detecting the presence or absence of biliary malignancies compared with currently available standard imaging techniques. The technique may be useful to evaluate biliary obstruction of unknown cause.


Expression analysis of novel striatal-enriched genes in Huntington disease.
Selective degeneration of striatal neurons is a pathologic hallmark of Huntington disease (HD). The exact mechanism(s) behind this specific neurodegeneration is still unknown. Expression studies of diseased human post-mortem brain, as well as different mouse models exhibiting striatal degeneration, have demonstrated changes in the expression of many important genes with a large proportion of changes being observed in the striatal-enriched genes. These investigations have raised questions about how enrichment of particular transcripts in the striatum can lead to its selective vulnerability to neurodegeneration. Monitoring the expression changes of striatal-enriched genes during the course of the disease may be informative about their potential involvement in selective degeneration. In this study, we analyzed a Serial Analysis of Gene Expression (SAGE) database (www.mouseatlas.org) and compared the mouse striatum to 18 other brain regions to generate a novel list of striatal-enriched transcripts. These novel striatal-enriched transcripts were subsequently evaluated for expression changes in the YAC128 mouse model of HD, and differentially expressed transcripts were further examined in human post-mortem caudate samples. We identified transcripts with altered expression in YAC128 mice, which also showed consistent expression changes in human post-mortem tissue. The identification of novel striatal-enriched genes with altered expression in HD offers new avenues of study, leading towards a better understanding of specific pathways involved in the selective degeneration of striatal neurons in HD.


Delayed activation of spinal microglia contributes to the maintenance of bone cancer pain in female Wistar rats via P2X7 receptor and IL-18.
Accumulating evidence suggests that activation of spinal microglia contributes to the development of inflammatory and neuropathic pain. However, the role of spinal microglia in the maintenance of chronic pain remains controversial. Bone cancer pain shares features of inflammatory and neuropathic pain, but the temporal activation of microglia and astrocytes in this model is not well defined. Here, we report an unconventional role of spinal microglia in the maintenance of advanced-phase bone cancer pain in a female rat model. Bone cancer elicited delayed and persistent microglial activation in the spinal dorsal horn on days 14 and 21, but not on day 7. In contrast, bone cancer induced rapid and persistent astrocytic activation on days 7-21. Spinal inhibition of microglia by minocycline at 14 d effectively reduced bone cancer-induced allodynia and hyperalgesia. However, pretreatment of minocycline in the first week did not affect the development of cancer pain. Bone cancer increased ATP levels in CSF, and upregulated P2X7 receptor, phosphorylated p38, and IL-18 in spinal microglia. Spinal inhibition of P2X7/p-38/IL-18 pathway reduced advanced-phase bone cancer pain and suppressed hyperactivity of spinal wide dynamic range (WDR) neurons. IL-18 induced allodynia and hyperalgesia after intrathecal injection, elicited mechanical hyperactivity of WDR neurons in vivo, and increased the frequency of mEPSCs in spinal lamina IIo nociceptive synapses in spinal cord slices. Together, our findings demonstrate a novel role of microglia in maintaining advanced phase cancer pain in females via producing the proinflammatory cytokine IL-18 to enhance synaptic transmission of spinal cord nociceptive neurons.


Conditional Release Placements of Insanity Acquittees in Oregon: 2012-2014.
Between January 1, 2012 and December 31, 2014, there was a large population (N = 200) of insanity acquittees placed on conditional release (CR) in the state of Oregon. This article looks at the demographic and system characteristics of this large group of individuals. The authors then focus on the initial housing placement and what happens to individuals after their release in relation to their housing placement. In Oregon, insanity acquittees are either conditionally released directly by the court or placed in the hospital prior to potential CR by a supervising board. In general, once CR occurs, individuals tend to stay in their initial placement without moving to less structured levels of care, raising concerns about transinstitutionalization. This is especially true for individuals released to the most structured living arrangement (secure residential treatment facility). Those individuals who are conditionally released to less structured settings have a higher rate of revocation back to the hospital. Those individuals who do move to less structured levels of care usually have longer hospital stays and start off in more structured levels of care to start their CR. Copyright © 2016 John Wiley & Sons, Ltd.


Reciprocal recognition of an HLA-Cw4-restricted HIV-1 gp120 epitope by CD8+ T cells and NK cells.
The HIV-1 Nef protein selectively downregulates human leukocyte antigen (HLA)-A and HLA-B but not HLA-C molecules on the surface of infected cells. This allows HIV-infected cells to evade recognition by most cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. We investigated the recognition of an HLA-Cw4-restricted HIV-1 gp120 epitope SFNCGGEFF (SF9) and its variant SFNCGGEFL (SL9) by T cells and NK receptors. Recognition of HIV-1 gp120 peptides (SF9 and SL9) by T-cell clones was measured by staining with HLA-Cw4-peptide tetrameric complexes and cytolytic assays using target cell pulsed with either peptides. KIR2DL1 binding to these two peptides was measured using surface plasmon resonance and tetramer staining of an NK cell line. : CTLs could recognize SF9 better than the variant SL9, as shown by both tetramer staining and cytolytic assays. Intriguingly, an HLA-Cw4 tetramer folded with the 'escape' variant SL9 could bind to KIR2DL1 on NK cell lines with higher affinity than HLA-Cw4-SF9. The binding of KIR2DL1 to its ligand results in inhibition of NK cell function. Our results indicate that the HIV-1 gp120 variant peptide SL9 could potentially escape both from NK cell and CTL recognition by increasing its affinity for KIR2DL1 binding. These data suggest that HIV-1 can acquire mutations that are capable of escaping from both CTL and NK cell recognition, a phenomenon we have termed 'double escape'.


Chronic arthritis in goats caused by a retrovirus.
A virus was isolated from an adult goat with chronic arthritis and shown to belong to the retrovirus group by electron microscopy and biochemical methods. Inoculation of the virus into cesarean-derived specific-pathogen-free goats' kids produced arthritic lesions similar to those in the spontaneous disease. Vrus was reisolated from the experimentally induced lesions.


Translocation of a hydrocarbon fluorescent probe between Epstein-Barr virus and lymphoid cells: an assay for early events in viral infection.
Translocation of the hydrocarbon fluorescent probe diphenylhexatriene (DPH) between membranes was studied by fluorescence polarization (P) analysis. First, using a model system, the high P value (0.324) of DPH-labeled cholesterol/phosphatidylcholine liposomes and the low P value (0.157) of DPH-labeled phosphatidylcholine liposomes allowed detection of DPH translocation between interacting liposomes. This was monitored by the change in P in either direction. Early events during cell-virus interactions were similarly studied by monitoring DPH translocation. The P value of DPH-labeled Epstein-Barr Virus (EBV) was significantly higher (0.350-0.392) than the P value of DPH-labeled lymphoid cells (0.238-0.289). Hence, DPH translocation could be detected by changes in P following incubation of DPH-labeled EBV and nonlabeled cells. A marked decrease in P was observed after incubation of DPH-labeled EBV with either nonlabeled lymphoblastoid Raji cells or fresh human B lymphocytes. However, only a slight decrease in P was obtained when DPH-labeled EBV was incubated with either nonlabeled fresh human T lymphocytes or fresh T or B rabbit lymphocytes. Moreover, incubation of fresh human B lymphocytes with the purified C3 component of complement (a putative inhibitor for the EBV receptor) prior to the addition of DPH-labeled EBV abolished the observed decrease in the P value. Most of these experiments were carried out with both the P3HR-1 and the B95-8 strains of EBV. DPH translocation, as determined by fluorescence polarization analysis, is, therefore, measuring some early event during interaction of this enveloped virus and mammalian cells. The potential applicability of this technique to other viruses is illustrated by an experiment with Semliki Forest virus.


Efficacy of ampicillin trihydrate or ceftiofur hydrochloride for treatment of metritis and subsequent fertility in dairy cows.
Our objectives were to evaluate the efficacy of ampicillin trihydrate for the treatment of metritis in dairy cows compared with ceftiofur hydrochloride and the subsequent effects on pregnancy at first insemination (P/AI). Cows in the first 12 d in milk (DIM) with a uterine discharge score of 5 (watery, reddish or brownish discharge of foul smell) and rectal temperature <39.5°C were diagnosed with metritis based on the fetid discharge, and cows with metritis and rectal temperature ≥39.5°C were diagnosed as having puerperal metritis. Cows with metritis (n=528) were blocked by parity and type of metritis as fetid discharge or puerperal metritis and, within each block, assigned randomly to receive 11mg/kg of ampicillin (n=259) or 2.2mg/kg of ceftiofur (n=269) once daily for 5 d. Day of diagnosis of metritis was considered study d 1. A cohort of 268 cows without metritis was selected randomly at 12 DIM. Rectal temperature was measured in cows with metritis on study d 1 to 7, and 12, and vaginal discharge was scored on study d 5, 7, and 12. Metritis cure was characterized by vaginal discharge score of <5 or by vaginal discharge score of <5 and no fever. At 32±3 DIM, vaginal discharge was scored for diagnosis of purulent vaginal discharge. At 39±3 DIM, endometrial cytology was performed. At 53±3 and 67±3 DIM, ovaries were scanned to determine estrous cyclicity. Pregnancy was evaluated after the first AI. Cure of metritis based on vaginal discharge <5 was greater for ampicillin than ceftiofur on d 5 (37.1 vs. 25.2%) and 7 (57.2 vs. 46.3%), but not on d 12 (82.0 vs. 85.0%). Cure of metritis based on vaginal discharge <5 and no fever was greater for ampicillin than for ceftiofur only on d 7 (50.4 vs. 37.9%), but not on d 5 (23.1 vs. 17.6%) and 12 (66.1 vs. 67.4%). Cows with puerperal metritis had reduced cure compared with cows with fetid discharge on d 5 (30.5 vs. 12.8%), 7 (55.2 vs. 33.6%), and 12 (72.0 vs. 61.1%). The proportion of cows with fever on any day after therapy started did not differ between treatments. Fifty-three percent of cows with metritis based on fetid discharge developed fever after initiating antimicrobial therapy. Cows receiving ampicillin had less prevalence of purulent vaginal discharge than those treated with ceftiofur (57.7 vs. 67.8%), but they were both greater than cows without metritis (21.9%). Prevalence of cytological endometritis did not differ between ampicillin and ceftiofur (30.0 vs. 25.4%), but they were both greater than cows without metritis (14.5%). The proportion of estrous cyclic cows (75.0%) and P/AI did not differ among treatments (ampicillin=28.0% vs. ceftiofur=28.3% vs. without metritis=30.5%). Clinical cure was faster for ampicillin than for ceftiofur, but on study d 12 both treatments resulted in similar cure. Clinical cure was less for cows with puerperal metritis than for cows with fetid uterine discharge. Despite differences in uterine health, P/AI at the first insemination did not differ among treatments.


Sorafenib Resistance in Hepatocellular Carcinoma: The Relevance of Genetic Heterogeneity.
Despite advances in biomedicine, the incidence and the mortality of hepatocellular carcinoma (HCC) remain high. The majority of HCC cases are diagnosed in later stages leading to the less than optimal outcome of the treatments. Molecular targeted therapy with sorafenib, a dual-target inhibitor targeting the serine-threonine kinase Raf and the tyrosine kinases VEGFR/PDGFR, is at present the main treatment for advanced-stage HCC, either in a single or combinatory regimen. However, it was observed in a large number of patients that its effectiveness is hampered by drug resistance. HCC is highly heterogeneous, within the tumor and among individuals, and this influences disease progression, classification, prognosis, and naturally cellular susceptibility to drug resistance. This review aims to provide an insight on how HCC heterogeneity influences the different primary mechanisms of chemoresistance against sorafenib including reduced drug intake, enhanced drug efflux, intracellular drug metabolism, alteration of molecular targets, activation/inactivation of signaling pathways, changes in the DNA repair machinery, and negative balance between apoptosis and survival of the cancer cells. The diverse variants, mutations, and polymorphisms in molecules and their association with drug response can be a helpful tool in treatment decision making. Accordingly, the existence of heterogeneous biomarkers in the tumor must be considered to strengthen multi-target strategies in patient-tailored treatment.


Chronic VEGF blockade worsens glomerular injury in the remnant kidney model.
VEGF inhibition can promote renal vascular and parenchymal injury, causing proteinuria, hypertension and thrombotic microangiopathy. The mechanisms underlying these side effects are unclear. We investigated the renal effects of the administration, during 45 days, of sunitinib (Su), a VEGF receptor inhibitor, to rats with 5/6 renal ablation (Nx). Adult male Munich-Wistar rats were distributed among groups S+V, sham-operated rats receiving vehicle only; S+Su, S rats given Su, 4 mg/kg/day; Nx+V, Nx rats receiving V; and Nx+Su, Nx rats receiving Su. Su caused no change in Group S. Seven and 45 days after renal ablation, renal cortical interstitium was expanded, in association with rarefaction of peritubular capillaries. Su did not worsen hypertension, proteinuria or interstitial expansion, nor did it affect capillary rarefaction, suggesting little angiogenic activity in this model. Nx animals exhibited glomerulosclerosis (GS), which was aggravated by Su. This effect could not be explained by podocyte damage, nor could it be ascribed to tuft hypertrophy or hyperplasia. GS may have derived from organization of capillary microthrombi, frequently observed in Group Nx+Su. Treatment with Su did not reduce the fractional glomerular endothelial area, suggesting functional rather than structural cell injury. Chronic VEGF inhibition has little effect on normal rats, but can affect glomerular endothelium when renal damage is already present.


[Continuing eco-pathological survey: 5. Demonstration of pathologic associations in dairy cow stock: individual data].
The calculation of pathological association ratio (or relative risk) from two ways contingency tables, allowed to show numerous interrelationships between diseases of 1,205 dairy cows. The analysis concerns the animal during productive life, the lactation (all lactation mixed), a particular one (1st, 2nd, 3rd and more) and the beginning of productive life of cow (three first lactations). The relationships are discussed according to the level of calculation (animal, lactation, or beginning of life). There is some triangular relations: calving trilogy (dystocia-stillbirth-retained placenta), infectious trilogy (foul of the foot-metritis-mastitis), metabolic trilogy (appetite disorders-ketosis-digestive disorders), foot disorders trilogy (lameness-non infectious diseases of foot-foul of the foot). These pathological groups seem to depend from common risk factors (climate-feeding-cowshed hygiene-control of heat-age-culling policy-genetic factors-anatomical or physiological predisposition, animal size...), but, numerous interactions between groups are also shown, leading to consider pathology in farm like a complex entity.


Spinal opioid receptor-sensitive muscle afferents contribute to the fatigue-induced increase in intracortical inhibition in healthy humans.
We investigated the influence of spinal opioid receptor-sensitive muscle afferents on cortical changes following fatiguing unilateral knee-extensor exercise. On separate days, seven subjects performed an identical five sets of intermittent isometric right-quadriceps contractions, each consisting of eight submaximal contractions [63 ± 7% maximal voluntary contraction (MVC)] and one MVC. The exercise was performed following either lumbar interspinous saline injection or lumbar intrathecal fentanyl injection blocking the central projection of spinal opioid receptor-sensitive lower limb muscle afferents. To quantify exercise-induced peripheral fatigue, quadriceps twitch force (Q(tw,pot)) was assessed via supramaximal magnetic femoral nerve stimulation before and after exercise. Motor evoked potentials and cortical silent periods (CSPs) were evaluated via transcranial magnetic stimulation of the motor cortex during a 3% MVC pre-activation period immediately following exercise. End-exercise quadriceps fatigue was significant and similar in both conditions (Q(tw,pot) -35 and -39% for placebo and fentanyl, respectively; P = 0.38). Immediately following exercise on both days, motor evoked potentials were similar to those obtained prior to exercise. Compared with pre-exercise baseline, CSP in the placebo trial was 21 ± 5% longer postexercise (P < 0.01). In contrast, CSP following the fentanyl trial was not significantly prolonged compared with the pre-exercise baseline (6 ± 4%). Our findings suggest that the central effects of spinal opioid receptor-sensitive muscle afferents might facilitate the fatigue-induced increase in CSP. Furthermore, since the CSP is thought to reflect inhibitory intracortical interneuron activity, which may contribute to central fatigue, our findings imply that spinal opioid receptor-sensitive muscle afferents might influence central fatigue by facilitating intracortical inhibition.


Associations Between Hand Hygiene Education and Self-Reported Hand-Washing Behaviors Among Korean Adults During MERS-CoV Outbreak.
Hand washing is an effective way to prevent transmission of infectious diseases. Education and promotional materials about hand washing may change individuals' awareness toward hand washing. Infectious disease outbreak may also affect individuals' awareness. Our study aimed to examine associations between hand-washing education and self-reported hand-washing behaviors among Korean adults during the year of the Middle East respiratory syndrome (MERS) outbreak. Data from the 2015 Community Health Survey were used for this study. The total study population comprised 222,599 individuals who were older than 20 years of age. A multiple linear regression model was used to investigate associations between hand hygiene education and self-reported hand-washing behaviors. Subgroup analyses stratified by age, sex, income, and MERS outbreak regions were also performed. Individuals who received hand-washing education or saw promotional materials related to hand washing had significantly higher scores for self-reported use of soap or sanitizer (β = 0.177, P < .0001) and self-reported frequency of hand washing (β = 0.481, P < .0001) than those who did not have such experiences. The effect of hand-washing education on self-reported behavior change was greater among older adults, women, and lower income earners. The effect of hand hygiene education on self-reported use of soap or sanitizer was similar regardless of whether the participants lived in MERS regions. Our findings emphasize the importance of education or promotions encouraging hand washing, especially for older adults, women, and lower income earners. In addition, MERS outbreak itself affected individuals' awareness of hand-washing behaviors. Well-organized campaigns that consider these factors are needed to prevent infectious diseases.


Dacryoadenitis associated with Acanthamoeba keratitis.
To report the incidence of dacryoadenitis as a symptom associated with Acanthamoeba keratitis. We investigated all cases of Acanthamoeba keratitis (20 patients and 21 eyes) diagnosed and treated at Tokyo Dental College, Ichikawa General Hospital, Ichikawa, Japan, between May 1, 1994, and November 30, 2005. We recorded the incidence of dacryoadenitis diagnosed using clinical signs of lacrimal gland swelling and pain on pressure, computed tomography, magnetic resonance imaging, and histopathologic analysis. Eight eyes (38%) of 8 patients had dacryoadenitis simultaneously with Acanthamoeba keratitis. Dacryoadenitis was diagnosed using histopathologic analysis and computed tomography in 1 patient, histopathologic analysis and magnetic resonance imaging in 1, magnetic resonance imaging in 2, and clinical signs alone in 4. Histopathologic examination in 2 patients revealed moderate infiltration of lymphocytes and plasma cells in the lacrimal gland compatible with dacryoadenitis. No Acanthamoeba organisms were found in the lacrimal gland. The standard protocol for Acanthamoeba keratitis was performed without particular treatment of dacryoadenitis in all patients. Lacrimal gland swelling improved after a mean of 10 weeks (range, 4-17 weeks) in conjunction with symptoms of keratitis; however, 1 patient (patient 1) required levator muscle surgery and blepharoplasty for residual ptosis. Dacryoadenitis is a clinical finding associated with Acanthamoeba keratitis.


Physical activity and 3-year BMI change in overweight and obese children.
Targeting physical activity (PA) is a mainstay in obesity treatment, but its BMI benefits are poorly quantified. We studied long-term predictive PA-BMI relationships in overweight/obese children presenting to primary care. Three-year follow-up of 182 overweight/obese 5- to 10-year-olds recruited from 45 Melbourne general practices. 7-day accelerometry (counts per minute, cpm). change in BMI z score, BMI category, and clinically significant BMI improvement (z score change ≥0.5). Linear and logistic regression. Mean (SD) baseline and 3-year BMI z scores were 1.8 (0.6) and 1.8 (0.7), and mean (SD) activity scores 334 (111) and 284 (104) cpm, respectively. Baseline activity did not predict BMI change. However, for every 100 cpm increase in change in activity over 3 years, BMI z score fell by 0.11 (95% confidence interval [CI] 0.03-0.20; P = .006). There were also trends toward greater odds of staying in the same, versus moving to a higher, BMI category (odds ratio 1.85, 95% CI 0.99-3.46) and clinically significant BMI improvement (odds ratio 1.96, 95% CI 0.90-4.27; P = .09). Change in percentage time spent in moderate-vigorous (P = .01), but not sedentary (P = .39) or light (P = .59), activity predicted reduced BMI z score. Sustained increase in moderate-vigorous PA predicts reducing BMI z score over 3 years in overweight/obese children presenting to primary care. However, the small BMI change associated with even the largest activity changes may explain disappointing BMI outcomes of brief primary care interventions targeting PA.


Role of nerve growth factor in the development of rat sympathetic neurons in vitro. II. Developmental studies.
Adrenergic sympathetic neurons were grown for 4 wk in submaximal and saturating concentrations of nerve growth factor (NGF) in the virtual absence of non-neuronal cells. In 0.2 or 5 microgram/ml 7S NGF, the neurons gradually decreased in number during the first week, although fewer neurons died at the higher level. No significant change in cell number was observed thereafter. Total neuronal protein, a measure of cell growth, increased linearly with age in both concentrations of NGF. At each age, neurons in high NGF exhibited greater growth per cell than those in low NGF. The ability of neurons to produce catecholamine (CA) increased dramatically during the second and third weeks in both concentrations of NGF, and along a similar time-course, although neurons in submaximal NGF developed a lesser capacity for CA production. As neurons developed in culture, they became less dependent on NGF for survival and CA production, but even in older cultures, approximately 50% of the neurons died when NGF was withdrawn.


Validation of a HPLC method for flavonoid biomarkers in skullcap (Scutellaria) and its use to illustrate wide variability in the quality of commercial tinctures.
To compare the flavonoid biomarker content (baicalin, baicalein and wogonin) of eleven commercial tinctures derived from Scutellaria lateriflora aerial parts (n=7) and Scutellaria baicalensis root (n=4). S. lateriflora tinctures are used in by western herbal practitioners to treat anxiety whereas S. baicalensis tinctures are used to treat inflammatory disease. Baicalin and baicalein were purchased from Aldrich Chemical Co. and Wogonin was purchased from ChromaDex. The internal standard (4-hydroxybenzoic acid) was obtained from Acros Organics. The column used was a Luna C18, 5 m (150 x 4.6 mm, Phenomenex) maintained at ambient room temperature. A HP1050 HPLC system was used, comprising a gradient pump with degasser, a variable wavelength UV detector set to 270 nm, and an autosampler. Gradient elution was performed using 0.1% formic acid (eluent A) and methanol (eluent B). The gradient elution initial conditions were 45% B with linear gradient to 60% from 2 to 10 min, followed by linear gradient to 70% B at 30 min, and then linear gradient to 99% B at 31 min, this proportion being maintained for 1 min. The mobile phase was then returned to initial conditions at 33 min and maintained until the end of the run at 35 min. The flow rate was 1 mL/min. The assay was validated for sensitivity, accuracy and reproducibility. The concentration range of biomarkers (baicalin, baicalein and wogonin) in commercial tinctures is reported for S. lateriflora (baicalin: 0-12.66 mg/mL; baicalein: 0-0.63 mg/mL; wogonin: 0-0.16 mg/mL) and for S. baicalensis (baicalin: 0.12-10.61 mg/mL; baicalein: 0.52-5.88 mg/mL; wogonin: 0.08-1.61 mg/mL). The wide variability in biomarker concentrations between commercial tinctures has important implications for the manufacturers of commercial tinctures, for herbal practitioners in the choice of tinctures and not least for pharmacology and clinical researchers.


The effect of age on the area of complete spatial summation for chromatic and achromatic stimuli.
Previously, an association between the area of complete spatial summation (Ricco's area) and age under scotopic conditions had been found. The authors sought to determine whether Ricco's area is similarly associated with age under photopic achromatic and selective S-cone conditions in peripheral vision and whether any association relates to a loss of ganglion cell density as determined by measurements of peripheral grating resolution acuity. Achromatic spatial summation functions were plotted for 68 healthy subjects (aged 20-77 years) in four oblique meridians on a gray background field of 10 cd/m(2). Similar functions were generated for the S-cone pathway (isolated using Stiles' two-color threshold method) for the same locations. Ricco's area was determined using two-phase regression analysis. Achromatic peripheral grating resolution acuity was measured at the same locations using high-contrast Gabor stimuli, as an estimate of localized functional ganglion cell density. There was a notable decrease in overall contrast sensitivity with age for all stimulus sizes. However, there was no evidence of age-related change in Ricco's area for either achromatic (superior field, r(2) = 0.05; inferior field, r(2) = 0.0007; all P > 0.05) or chromatic (superior field, r(2) = 0.01; inferior field, r(2) = 0.006; all P > 0.05) stimuli, despite a significant decrease in peripheral grating resolution acuity with age (superior field, r(2) = 0.15; inferior field, r(2) = 0.17; both P < 0.05). An age-related decline in functional ganglion cell density is not accompanied by a significant change in Ricco's area for achromatic or chromatic stimuli.