Orbiviruses in the Mediterranean basin: updated epidemiological situation of Bluetongue and new methods for the detection of BTV serotype 4.
Bluetongue virus (BTV), a member of the Orbivirus genus, has contributed to great economical losses in countries across the Mediterranean basin. Although BTV has an African origin, it has been reported in the South of Europe since 1924 when it was first detected in Cyprus. After this first Bluetongue (BT) outbreak, many others followed in most Mediterranean countries, resulting in seven BTV serotypes detected in 17 countries in Central-, South-Europe and North Africa in the last 10 years. Currently, six BTV serotypes (1, 2, 4, 8, 9 and 16) are circulating throughout Central- and Western Europe. The unexpected occurrence of BTV serotype 8 in Central Europe in 2006 and its spread and persistence have evidenced changes in the BTV scenario: new serotypes, never detected in the Mediterranean area so far, are playing a central role in the maintenance of BTV in Europe, and new species of Culicoides are now confirmed to be able to transmit the virus. Therefore, it is necessary to improve and implement specific assays in order to identify the serotypes currently present in different Mediterranean countries in a fast and reliable way. In this study, we present a new gel-based and real-time RT-PCR assay for the detection of BTV serotype 4. The sequence amplified in this test is located within BTV segment 2, a variable region of BTV dsRNA genome encoding the major outer-capsid protein VP2. No cross-reaction has been shown with other genetically or geographically related viruses and the sensitivity of this test allows the detection of 1.5-15 TCID50/ml of BTV.


An immunohistochemical and prognostic evaluation of cathepsin D expression in 105 bladder carcinomas.
Cathepsin D is a widely expressed aspartyl lysosomal protease. Clinical studies in several tumor types have shown a strong correlation between cathepsin D expression and tumor progression. In breast carcinoma, its expression is an independent prognostic factor associated with an increased risk of death. However, there have been no studies evaluating cathepsin D in bladder tumors. Therefore, the aim of this study was to determine the pattern of expression of cathepsin D in a large series of bladder carcinomas and assess its role as a prognostic factor against established variables. The tumors from 105 patients (median age 73) (median follow-up 26 months) with transitional cell carcinoma of bladder were examined. Forty-nine patients had superficial tumors (16 pTa; 33 pT1), 56 had invasive tumors (14 pT2; 42 pT3); there were 35 grade 1/2 tumors and 70 grade 3 tumors. These were stained by a standard immunohistochemical technique with an anti-cathepsin D monoclonal antibody. All 4 normal bladder specimens were positive for cathepsin D. Fifty-four tumors (51%) were positive for cathepsin D and 51 (49%) were negative. Chi square analysis showed a significant positive relationship between negative cathepsin D expression and stage (p < 0.0005), grade (p < 0.0001) and tumor morphology (p = 0.001). There was no relationship between cathepsin D expression and tumor ploidy (p > 0.1) or patient age (p = 0.09). Univariate analysis of disease-free and overall survival showed that negative cathepsin D expression (p = 0.01 and p = 0.0003 respectively), stage (p = 0.004 and p < 0.005 respectively) and grade (p = 0.02 and p = 0.0007 respectively) were associated with significantly worse prognosis. However, in a multivariate analysis of age, stage, grade and cathepsin D expression, only stage remained significant for overall survival (p < 0.005). The observed result for cathepsin D in the univariate analysis is probably due to its strong association with grade and stage. Nevertheless, cathepsin D status was able to provide additional prognostic information for overall survival in invasive tumors when stratifying for grade (p = 0.047), which suggests that it might provide additional prognostic data within particular tumor stages.


High-order myopic coronagraphic phase diversity (COFFEE) for wave-front control in high-contrast imaging systems.
The estimation and compensation of quasi-static aberrations is mandatory to reach the ultimate performance of high-contrast imaging systems. COFFEE is a focal plane wave-front sensing method that consists in the extension of phase diversity to high-contrast imaging systems. Based on a Bayesian approach, it estimates the quasi-static aberrations from two focal plane images recorded from the scientific camera itself. In this paper, we present COFFEE's extension which allows an estimation of low and high order aberrations with nanometric precision for any coronagraphic device. The performance is evaluated by realistic simulations, performed in the SPHERE instrument framework. We develop a myopic estimation that allows us to take into account an imperfect knowledge on the used diversity phase. Lastly, we evaluate COFFEE's performance in a compensation process, to optimize the contrast on the detector, and show it allows one to reach the 10(-6) contrast required by SPHERE at a few resolution elements from the star. Notably, we present a non-linear energy minimization method which can be used to reach very high contrast levels (better than 10(7) in a SPHERE-like context).


Caloric restriction reveals a metabolomic and lipidomic signature in liver of male mice.
Lipid composition, particularly membrane unsaturation, has been proposed as being a lifespan determinant, but it is currently unknown whether caloric restriction (CR), an accepted life-extending intervention, affects cellular lipid profiles. In this study, we employ a liquid chromatography quadrupole time-of-flight-based methodology to demonstrate that CR in the liver of male C57BL/6 mice: (i) induces marked changes in the cellular lipidome, (ii) specifically reduces levels of a phospholipid peroxidation product, 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphatidylcholine, (iii) alters cellular phosphoethanolamine and triglyceride distributional profiles, (iv) affects mitochondrial electron transport chain complexes, increasing complex II and decreasing complex III and (v) is associated with specific changes in liver metabolic pathways. These data demonstrate that CR induces a specific lipidome and metabolome reprogramming event in mouse liver which is associated with lower protein oxidative damage, as assessed by mass spectrometry-based measurements. Such changes may be critical to the increased lifespan and healthspan observed in C57BL/6 mice following CR.


Carbohydrate-binding agents cause deletions of highly conserved glycosylation sites in HIV GP120: a new therapeutic concept to hit the achilles heel of HIV.
Mannose-binding proteins derived from several plants (i.e. Hippeastrum hybrid and Galanthus nivalis agglutinin) or prokaryotes (i.e. cyanovirin-N) inhibit human immunodeficiency virus (HIV) replication and select for drug-resistant viruses that show profound deletion of N-glycosylation sites in the GP120 envelope (Balzarini, J., Van Laethem, K., Hatse, S., Vermeire, K., De Clercq, E., Peumans, W., Van Damme, E., Vandamme, A.-M., Bolmstedt, A., and Schols, D. (2004) J. Virol. 78, 10617-10627; Balzarini, J., Van Laethem, K., Hatse, S., Froeyen, M., Van Damme, E., Bolmstedt, A., Peumans, W., De Clercq, E., and Schols, D. (2005) Mol. Pharmacol. 67, 1556-1565). Here we demonstrated that the N-acetylglucosamine-binding protein from Urtica dioica (UDA) prevents HIV entry and eventually selects for viruses in which conserved N-glycosylation sites in GP120 were deleted. In contrast to the mannose-binding proteins, which have a 50-100-fold decreased antiviral activity against the UDA-exposed mutant viruses, UDA has decreased anti-HIV activity to a very limited extent, even against those mutant virus strains that lack at least 9 of 22 ( approximately 40%) glycosylation sites in their GP120 envelope. Therefore, UDA represents the prototype of a new conceptual class of carbohydrate-binding agents with an unusually specific and targeted drug resistance profile. It forces HIV to escape drug pressure by deleting the indispensable glycans on its GP120, thereby obligatorily exposing previously hidden immunogenic epitopes on its envelope.


Imaging of Abductor Tears: Stepwise Technique for Accurate Diagnosis.
This article presents the authors' technique for evaluation of possible abductor tears with magnetic resonance imaging (MRI). By using T2 fat-saturated coronal and axial slices of the hip and T1 coronal images of the pelvis, the authors show how to visualize all tears of the abductor tendons and signs of chronic abductor deficiency. After diagnosis has been made, signs that may help the surgeon determine if open or arthroscopic surgery would be best used are reviewed. Finally, the author reviews indications for a gluteus medius repair with concomitant gluteus maximus reconstruction.


Eccentric exercise activates novel transcriptional regulation of hypertrophic signaling pathways not affected by hormone changes.
Unaccustomed eccentric exercise damages skeletal muscle tissue, activating mechanisms of recovery and remodeling that may be influenced by the female sex hormone 17beta-estradiol (E2). Using high density oligonucleotide based microarrays, we screened for differences in mRNA expression caused by E2 and eccentric exercise. After random assignment to 8 days of either placebo (CON) or E2 (EXP), eighteen men performed 150 single-leg eccentric contractions. Muscle biopsies were collected at baseline (BL), following supplementation (PS), +3 hours (3H) and +48 hours (48H) after exercise. Serum E2 concentrations increased significantly with supplementation (P<0.001) but did not affect microarray results. Exercise led to early transcriptional changes in striated muscle activator of Rho signaling (STARS), Rho family GTPase 3 (RND3), mitogen activated protein kinase (MAPK) regulation and the downstream transcription factor FOS. Targeted RT-PCR analysis identified concurrent induction of negative regulators of calcineurin signaling RCAN (P<0.001) and HMOX1 (P = 0.009). Protein contents were elevated for RND3 at 3H (P = 0.02) and FOS at 48H (P<0.05). These findings indicate that early RhoA and NFAT signaling and regulation are altered following exercise for muscle remodeling and repair, but are not affected by E2.


Natural dyes versus lysochrome dyes in cheiloscopy: A comparative evaluation.
Cheiloscopy is the study of lip prints. Lip prints are genotypically determined and are unique, and stable. At the site of crime, lip prints can be either visible or latent. To develop lip prints for study purpose various chemicals such as lysochrome dyes, fluorescent dyes, etc. are available which are very expensive. Vermilion (Sindoor used by married Indian women) and indigo dye (fabric whitener) are readily available, naturally derived, and cost-effective reagents available in India. To compare the efficacy of sudan black, vermilion, and indigo in developing visible and latent lip prints made on bone china cup, satin fabric, and cotton fabric. Out of 45 Volunteers 15 lip prints were made on bone China cup 15 lip prints on Satin fabric and 15 on Cotton fabric. Sudan black, vermilion and indigo were applied on visible and latent lip prints and graded as good (+,+), fair (+), and poor (-) and statistically evaluated. The vermilion and indigo dye gives comparable results to that of sudan black for developing visible and latent lip prints.


Comparison of systematic transrectal biopsy to transperineal magnetic resonance imaging/ultrasound-fusion biopsy for the diagnosis of prostate cancer.
To compare targeted, transperineal magnetic resonance imaging (MRI)/ultrasound (US)-fusion biopsy to systematic transrectal biopsy in patients with previous negative or first prostate biopsy and to evaluate the gain in diagnostic information with systematic biopsies in addition to targeted MRI/US-fusion biopsies. In all, 263 consecutive patients with suspicion of prostate cancer were investigated. All patients were evaluated by 3-T multiparametric MRI (mpMRI) applying the European Society of Urogenital Radiology criteria. All patients underwent MRI/US-fusion biopsy transperineally (mean nine cores) and additionally a systematic transrectal biopsy (mean 12 cores). In all, 195 patients underwent repeat biopsy and 68 patients underwent first biopsy. The median age was 66 years, median PSA level was 8.3 ng/mL and median prostate volume was 50 mL. Overall, the prostate cancer detection rate was 52% (137/263). MRI/US-fusion biopsy detected significantly more cancer than systematic prostate biopsy (44% [116/263] vs 35% [91/263]; P = 0.002). In repeat biopsy, the detection rate was 44% (85/195) in targeted and 32% (62/195) in systematic biopsy (P = 0.002). In first biopsy, the detection rate was 46% (31/68) in targeted and 43% (29/68) in systematic biopsy (P = 0.527). In all, 80% (110/137) of biopsy confirmed prostate cancers were clinically significant. For the upgrading of Gleason score, 44% (32/72) more clinically significant prostate cancer was detected by using additional targeted biopsy than by systematic biopsy alone. Conversely, 12% (10/94) more clinically significant cancer was found by systematic biopsy additionally to targeted biopsy. MRI/US-fusion biopsy was associated with a higher detection rate of clinically significant prostate cancer while taking fewer cores, especially in patients with prior negative biopsy. Due to a high portion of additional tumours with Gleason score ≥7 detected in addition to targeted biopsy, systematic biopsy should still be performed additionally to targeted biopsy.


Breakup of Finite-Size Colloidal Aggregates in Turbulent Flow Investigated by Three-Dimensional (3D) Particle Tracking Velocimetry.
Aggregates grown in mild shear flow are released, one at a time, into homogeneous isotropic turbulence, where their motion and intermittent breakup is recorded by three-dimensional particle tracking velocimetry (3D-PTV). The aggregates have an open structure with a fractal dimension of ∼2.2, and their size is 1.4 ± 0.4 mm, which is large, compared to the Kolmogorov length scale (η = 0.15 mm). 3D-PTV of flow tracers allows for the simultaneous measurement of aggregate trajectories and the full velocity gradient tensor along their pathlines, which enables us to access the Lagrangian stress history of individual breakup events. From this data, we found no consistent pattern that relates breakup to the local flow properties at the point of breakup. Also, the correlation between the aggregate size and both shear stress and normal stress at the location of breakage is found to be weaker, when compared with the correlation between size and drag stress. The analysis suggests that the aggregates are mostly broken due to the accumulation of the drag stress over a time lag on the order of the Kolmogorov time scale. This finding is explained by the fact that the aggregates are large, which gives their motion inertia and increases the time for stress propagation inside the aggregate. Furthermore, it is found that the scaling of the largest fragment and the accumulated stress at breakup follows an earlier established power law, i.e., dfrag ∼ σ(-0.6) obtained from laminar nozzle experiments. This indicates that, despite the large size and the different type of hydrodynamic stress, the microscopic mechanism causing breakup is consistent over a wide range of aggregate size and stress magnitude.


Store-operated Ca²⁺ entry and depolarization explain the anomalous behaviour of myometrial SR: effects of SERCA inhibition on electrical activity, Ca²⁺ and force.
In the myometrium SR Ca(2+) depletion promotes an increase in force but unlike several other smooth muscles, there is no Ca(2+) sparks-STOCs coupling mechanism to explain this. Given the importance of the control of contractility for successful parturition, we have examined, in pregnant rat myometrium, the effects of SR Ca(2+)-ATPase (SERCA) inhibition on the temporal relationship between action potentials, Ca(2+) transients and force. Simultaneous recording of electrical activity, calcium and force showed that SERCA inhibition, by cyclopiazonic acid (CPA 20 μM), caused time-dependent changes in excitability, most noticeably depolarization and elevations of baseline [Ca(2+)]i and force. At the onset of these changes there was a prolongation of the bursts of action potentials and a corresponding series of Ca(2+) spikes, which increased the amplitude and duration of contractions. As the rise of baseline Ca(2+) and depolarization continued a point was reached when electrical and Ca(2+) spikes and phasic contractions ceased, and a maintained, tonic force and Ca(2+) was produced. Lanthanum, a non-selective blocker of store-operated Ca(2+) entry, but not the L-type Ca(2+) channel blocker nifedipine (1-10 μM), could abolish the maintained force and calcium. Application of the agonist, carbachol, produced similar effects to CPA, i.e. depolarization, elevation of force and calcium. A brief, high concentration of carbachol, to cause SR Ca(2+) depletion without eliciting receptor-operated channel opening, also produced these results. The data obtained suggest that in pregnant rats SR Ca(2+) release is coupled to marked Ca(2+) entry, via store operated Ca(2+) channels, leading to depolarization and enhanced electrical and mechanical activity.


Equilibrium ultrastable glasses produced by random pinning.
Ultrastable glasses have risen to prominence due to their potentially useful material properties and the tantalizing possibility of a general method of preparation via vapor deposition. Despite the importance of this novel class of amorphous materials, numerical studies have been scarce because achieving ultrastability in atomistic simulations is an enormous challenge. Here, we bypass this difficulty and establish that randomly pinning the position of a small fraction of particles inside an equilibrated supercooled liquid generates ultrastable configurations at essentially no numerical cost, while avoiding undesired structural changes due to the preparation protocol. Building on the analogy with vapor-deposited ultrastable glasses, we study the melting kinetics of these configurations following a sudden temperature jump into the liquid phase. In homogeneous geometries, we find that enhanced kinetic stability is accompanied by large scale dynamic heterogeneity, while a competition between homogeneous and heterogeneous melting is observed when a liquid boundary invades the glass at constant velocity. Our work demonstrates the feasibility of large-scale, atomistically resolved, and experimentally relevant simulations of the kinetics of ultrastable glasses.


Dyskerin expression in human fetal, adult and neoplastic intrahepatic bile ducts: correlations with cholangiocarcinoma aggressiveness.
To investigate the immunohistochemical expression of dyskerin, a biomarker involved in ribosome production and telomere maintenance, in human fetal, adult and neoplastic bile ducts, and possible correlations with cholangiocarcinoma aggressiveness. Sixty consecutive intrahepatic cholangiocarcinomas were collected and used for tissue microarray construction (total: 176 cores); clinical data and follow-up were also collected. Five fetal and 10 normal adult livers were included as controls. Automated immunohistochemistry for dyskerin, p53, and Ki67, and nucleolar silver staining, were performed. In normal livers, dyskerin expression was negative in smaller bile ducts (mean 44.8 μm) and positive in bile ducts of larger diameter (mean 116.1 μm; P < 0.001). Expression was positive in 56.7% of cholangiocarcinomas, and correlated with p53 mutation (P = 0.008) and a higher proliferative (Ki67) index (P = 0.003), which were included as markers of tumour aggressiveness. Finally, dyskerin-positive cholangiocarcinomas showed a negative trend in disease-free survival (P = 0.078) on univariate analysis. The non-neoplastic biliary tree seems to progressively lose dyskerin expression from the major branches to the peripheral portal bile ducts. Similarly, intrahepatic cholangiocarcinomas showed two patterns of dyskerin expression, and the dyskerin-positive phenotype seemed to characterize more aggressive cholangiocarcinomas.


Postoperative mortality after surgery for brain tumors by patient insurance status in the United States.
To examine whether being uninsured is associated with higher in-hospital postoperative mortality when undergoing surgery in the United States for a brain tumor. Retrospective cohort study using the Nationwide Inpatient Sample, January 1, 1999, through December 31, 2008. The Nationwide Inpatient Sample contains all inpatient records from a stratified sample of 20% of hospitals in 37 states. A total of 28,581 patients, aged 18 to 65 years, who underwent craniotomy for a brain tumor. Three groups were studied: Medicaid recipients and privately insured and uninsured patients. The main outcome measure was in-hospital postoperative death. Associations between this outcome and insurance status were examined within the full cohort and within the subset of patients with no comorbidity using Cox proportional hazards models. These models were stratified by hospital to control for any clustering effects that could arise from differing access to care. In the unadjusted analysis, the mortality rate for privately insured patients was 1.3% (95% CI, 1.1%-1.4%) compared with 2.6% for uninsured patients (95% CI, 1.9%-3.3%; P < .001) and 2.3% for Medicaid recipients (95% CI, 1.8%-2.8%; P < .001). After adjusting for patient characteristics and stratifying by hospital in patients with no comorbidity, uninsured patients still had a higher risk of experiencing in-hospital death (hazard ratio, 2.62; 95% CI, 1.11-6.14; P = .03) compared with privately insured patients. In this adjusted analysis, the disparity was not conclusively present in Medicaid recipients (hazard ratio, 2.03; 95% CI, 0.97-4.23; P = .06). Uninsured patients who underwent craniotomy for a brain tumor experienced the highest in-hospital mortality. Differences in overall health do not fully account for this disparity.


Is interleukin-4delta3 splice variant expression in bovine tuberculosis a marker of protective immunity?
Splice variants of the interleukin-4 (IL-4) cytokine gene have been described for humans, mice, and cattle. IL-4 splice variants have been shown to inhibit IL-4-mediated cellular responses and thus act as IL-4 antagonists. Recent work has highlighted the possibility of a correlation between IL-4 splice variants and protection against clinical tuberculosis. In this study we investigated the potential role of IL-4 splice variants IL-4delta2 and IL-4delta3 in cattle with bovine tuberculosis, using quantitative real-time reverse transcription-PCR. For this analysis we used naturally exposed tuberculin skin test-positive field reactor cattle, uninfected control cattle, and cattle from two experimental models of protective immunity against Mycobacterium bovis: (i) vaccination with M. bovis BCG and challenge with virulent M. bovis and (ii) infection with M. bovis and treatment with isoniazid (INH) prior to rechallenge. The cytokine levels of field reactor cattle were compared to the levels of uninfected controls, while in kinetic studies of BCG vaccination and INH treatment we compared pre-experimental values with sequential samples for each individual animal. The data revealed a significant increase in IL-4delta3 mRNA expression in field reactor cattle, which had no visible pathology compared to cattle with gross pathology typical of bovine tuberculosis. Increased IL-4delta3 expression in both cattle models of protective immunity (BCG vaccination and INH treatment) was transient over time, reaching significance in the INH treatment model. Our results support the hypothesis that IL-4delta3 is involved in protective immunity against M. bovis infection in cattle and are in accordance with clinical studies that have suggested a role for IL-4 splice variants in protective immunity in tuberculosis.


Meaningful feedback through a sociocultural lens.
This AMEE guide provides a framework and practical strategies for teachers, learners and institutions to promote meaningful feedback conversations that emphasise performance improvement and professional growth. Recommended strategies are based on recent feedback research and literature, which emphasise the sociocultural nature of these complex interactions. We use key concepts from three theories as the underpinnings of the recommended strategies: sociocultural, politeness and self-determination theories. We view the content and impact of feedback conversations through the perspective of learners, teachers and institutions, always focussing on learner growth. The guide emphasises the role of teachers in forming educational alliances with their learners, setting a safe learning climate, fostering self-awareness about their performance, engaging with learners in informed self-assessment and reflection, and co-creating the learning environment and learning opportunities with their learners. We highlight the role of institutions in enhancing the feedback culture by encouraging a growth mind-set and a learning goal-orientation. Practical advice is provided on techniques and strategies that can be used and applied by learners, teachers and institutions to effectively foster all these elements. Finally, we highlight throughout the critical importance of congruence between the three levels of culture: unwritten values, espoused values and day to day behaviours.


Paraurethral cyst in adult women: experience with 85 cases.
To present the clinical experience in the management of Skene's duct cysts (paraurethral cysts) in women. A retrospective chart review of patients who have underwent surgical treatment for paraurethral cyst between 2002 and 2012 was performed. A total of 85 women were diagnosed with paraurethral cyst over a 10-year period. The paraurethral cysts were detected at vaginal examination. Evaluations consisted of urine analysis and culture and urinary tract ultrasound. The first 20 cases underwent cystourethroscopy as well. All patients underwent surgical incision, drainage and marsupialization of the cyst. They were followed for evidence of any complications and recurrence. The mean follow up time was 5.5 years. Totally, 83 patients (97.6%) were cured. There were two cases of recurrence which were treated with second surgical attempt. Most paraurethral cysts in women may be diagnosed by history and physical examination alone. Simple incision and marsupialization of the female paraurethral cyst was effective in more than 97% of our patients, without recurrence.


The influence of stochastic temperature fluctuations in shaping the physiological performance of the California mussel, Mytilus californianus.
Climate change is forecasted to increase temperature variability and stochasticity. Most of our understanding of thermal physiology of intertidal organisms has come from laboratory experiments that acclimate organisms to submerged conditions and steady-state increases in temperatures. For organisms experiencing the ebb and flow of tides with unpredictable low tide aerial temperatures, the reliability of reported tolerances and thus predicted responses to climate change requires incorporation of environmental complexity into empirical studies. Using the mussel Mytilus californianus, our study examined how stochasticity of the thermal regime influences physiological performance. Mussels were acclimated to either submerged conditions or a tidal cycle that included either predictable, unpredictable or no thermal stress during daytime low tide. Physiological performance was measured through anaerobic metabolism, energy stores and cellular stress mechanisms just before low tide, and cardiac responses during a thermal ramp. Both air exposure and stochasticity of temperature change were important in determining thermal performance. Glycogen content was highest in the mussels from the unpredictable treatment, but there was no difference in the expression of heat shock proteins between thermal treatments, suggesting that mussels prioritise energy reserves to deal with unpredictable low tide conditions. Mussels exposed to fluctuating thermal regimes had lower gill anaerobic metabolism, which could reflect increased metabolic capacity. Our results suggest that although thermal magnitude plays an important role in shaping physiological performance, other key elements of the intertidal environment complexity such as stochasticity, thermal variability and thermal history are also important considerations for determining how species will respond to climate warming.


Typha capensis (Rohrb.)N.E.Br. (bulrush) extract scavenges free radicals, inhibits collagenase activity and affects human sperm motility and mitochondrial membrane potential in vitro: a pilot study.
The biodiversity in South Africa provides more than 30,000 higher plants, of which more than 3000 are used by traditional healers to treat diseases. Typha capensis (bulrush) is one of the medicinal plants used in South Africa to treat male fertility problems. Considering that South African traditional healers have been recognised by Law and the health benefits of T. capensis have not been scientifically investigated yet, this study aimed at investigating the in vitro effects of aqueous extracts from this plant on male reproductive functions. Both leaves and rhizomes of T. capensis were dried, infused with distilled water and freeze-dried. Motile sperm from 50 men were isolated by swim-up and incubated with 1 μg ml(-1) aqueous extract of Typha rhizome for 1 h at 37 °C. Vitality, motility, sperm production of reactive oxygen species and mitochondrial membrane potential were analysed in the test sample, a control and in the pellet from the swim-up. Results showed that the rhizome extract had significant (P < 0.0001) negative effects on all parameters. The extracts from the leaves and rhizomes revealed dose-dependent inhibitory activity for collagenase and free radical formation. No inhibitory activity for elastase was found. The inhibitory activity for collagenase might indicate possible anti-cancer effects.


Participation in a 10-week course of yoga improves behavioural control and decreases psychological distress in a prison population.
Yoga and meditation have been shown to be effective in alleviating symptoms of depression and anxiety in healthy volunteers and psychiatric populations. Recent work has also indicated that yoga can improve cognitive-behavioural performance and control. Although there have been no controlled studies of the effects of yoga in a prison population, we reasoned that yoga could have beneficial effects in a setting where psychosocial functioning is often low, and the frequency of impulsive behaviours is high. Participants were recruited from 7 British prisons and randomly allocated to either a 10-week yoga programme (yoga group; 1 class per week; N = 45) or a control group (N = 55). Self-report measures of mood, stress, and psychological distress were collected before and after the intervention period. Participants completed a cognitive-behavioural task (Go/No-Go) at the end of the study, which assessed behavioural response inhibition and sustained attention. Participants in the yoga group showed increased self-reported positive affect, and reduced stress and psychological distress, compared to participants in the control group. Participants who completed the yoga course also showed better performance in the cognitive-behavioural task, making significantly fewer errors of omission in Go trials and fewer errors of commission on No-Go trials, compared to control participants. Yoga may be effective in improving subjective wellbeing, mental health, and executive functioning within prison populations. This is an important consideration given the consistently high rates of psychological morbidity in this group and the need for effective and economical intervention programmes.


[Comparative studies on codon usage bias of Ganoderma lucidum based on analysis of genomic and transcriptomic data].
Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.


[A successful multimodality therapy for a case of recurrent rectal cancer with KRAS mutation].
A man in his 50s underwent high anterior resection for rectosigmoid cancer in January 2010. The primary tumor was diagnosed as a moderately differentiated adenocarcinoma with KRAS mutation, pStage III a. In May 2011, the patient had a recurrent lung tumor detected by computed tomography(CT); the tumor was resected using video-assisted thoracoscopic surgery. However, additional recurrent lung tumors arose, and radiofrequency ablation (RFA) was performed to treat these in February 2012. After RFA therapy, capecitabine was administered as adjuvant chemotherapy. Unfortunately, 10 months later, positron emission tomography (PET) /CT suggested a new recurrence in a left lateral lymph node. Although the pelvic lymph node was surgically removed immediately, a new lung recurrence was found on CT three months after the surgery. RFA was again used to treat this lung lesion. After the second RFA, the patient is doing well without any evidence of recurrence. We describe a case of recurrent rectal cancer successfully treated with multimodality therapy. The combination of appropriate local therapy with systemic chemotherapy is an essential strategy to treat advanced colorectal cancer, especially in patients with KRAS mutation when anti-EGFR antibodies are not effective.


Introduction: Forecasting Informatics Competencies for Nurses in the Future of Connected Health.
This introduction to the book discusses how the topic of competencies for nurses in a world of connected health needs to be addressed at the curriculum level to achieve the specific competencies for various roles, including practicing nurse, nurse teacher, nurse leader, and nursing informatics specialists. It looks back at milestone publications from the international Nursing Informatics post conferences that still serve a purpose for inspiring developments today and looks forward to the way nurses can use connected health to improve the health and health care for their patients. Specific emerging topics in health information technology are addressed as well, such as semantics, genetics, big data, eHealth and social media.


Mean platelet volume is elevated in patients with psoriasis vulgaris.
This retrospective study was done to investigate the mean platelet volume (MPV) level in patients with psoriasis vulgaris and its relationship with disease severity. We undertook a cross-sectional study on 176 patients and 101 healthy controls to examine the association between MPV and psoriasis. Various clinical and laboratory parameters were analyzed and compared. Platelet distribution width and MPV were significantly higher in patients with psoriasis than controls. In addition, there was positive correlation between Psoriasis Area Severity Index (PASI) and MPV. When psoriasis patients were grouped into mild psoriasis (PASI<10) and moderate to severe psoriasis (PASI≥10), the MPV of the latter group was significantly elevated. Nevertheless, patients with higher MPV level (MPV≥10.4 fL) did not show higher PASI than lower MPV level (MPV<10.4 fL). MPV levels significantly decreased after improvements of psoriasis with various treatments. The variations of MPV and PASI also showed significant correlation. We have shown that MPV is increased in psoriasis patients and correlates with disease severity. Therefore, MPV levels may be considered as a marker of disease severity of psoriasis.


New findings for phenotype-genotype correlations in a large European series of holoprosencephaly cases.
Holoprosencephaly (HPE) is the most common forebrain defect in humans. It results from incomplete midline cleavage of the prosencephalon. A large European series of 645 HPE probands (and 699 relatives), consisting of 51% fetuses and 49% liveborn children, is reported. Mutations in the four main genes involved in HPE (SHH, ZIC2, SIX3, TGIF) were identified in 25% of cases. The SHH, SIX3, and TGIF mutations were inherited in more than 70% of these cases, whereas 70% of the mutations in ZIC2 occurred de novo. Moreover, rearrangements were detected in 22% of the 260 patients screened by array comparative genomic hybridisation. 15 probands had two mutations providing additional support for the 'multiple-hit process' in HPE. There was a positive correlation between the severity of the brain malformation and facial features for SHH, SIX3, and TGIF, but no such correlation was found for ZIC2 mutations. The most severe HPE types were associated with SIX3 and ZIC2 mutations, whereas microforms were associated with SHH mutations. The study focused on the associated brain malformations, including neuronal migration defects, which predominated in individuals with ZIC2 mutations, and neural tube defects, which were frequently associated with ZIC2 (rachischisis) and TGIF mutations. Extracraniofacial features were observed in 27% of the individuals in this series (up to 40% of those with ZIC2 mutations) and a significant correlation was found between renal/urinary defects and mutations of SHH and ZIC2. An algorithm is proposed based on these new phenotype-genotype correlations, to facilitate molecular analysis and genetic counselling for HPE.


First Report of Soybean vein necrosis virus on Soybeans in Michigan.
Soybean vein necrosis virus (SVNV) is associated with an emerging disease in soybean producing regions of the United States. Soybean leaves with necrotic vein symptoms were initially noted in 2008 or 2009 in fields across Arkansas, Kansas, Missouri, Illinois, Mississippi, Tennessee, and Kentucky and SVNV was determined to be the causal agent (2). In 2012, widespread reports of SVNV were made across most soybean (Glycine max) producing states including the recent confirmation of SVNV in Iowa and Wisconsin (1). Foliar symptoms similar to those reported for SVNV were observed at approximately 1 to 10% incidence in soybean fields across Michigan in late August and September of 2012, including fields located in Cass, Ingham, Midland, Saginaw, and Van Buren counties. Symptoms included chlorosis and necrosis which initiated on the veins with subsequent spread across the leaf. An initial sample collected from the East Lansing Agricultural Research Station was confirmed to have SVNV with a polyclonal antibody using double antibody sandwich (DAS)-ELISA at AC Diagnostics, Inc. (Fayetteville, AR) and with reverse transcription PCR by Ioannis Tzanetakis, University of Arkansas, Fayetteville. Additional samples from five fields were subsequently collected from Cass, Ingham, and Van Buren counties. Duplicate leaf tissue samples were tested with DAS-ELISA using the SVNV test kit (AC Diagnostics). All symptomatic leaf samples exhibited a strong positive reaction based on the optical density reading at 405 nm. Absorbance reading that exceeded the healthy soybean tissue by a standard deviation of +3× were considered positive. Total RNA was also extracted from each sample using the RNeasy Plant Mini Kit (Qiagen, Germantown, MD). Complementary DNA (cDNA) was generated using virus-specific LdetR and SdetR primers (2) with the High Capacity RT cDNA kit (Life Technologies; Carlsbad, CA). The cDNA was used as template for PCR with the SVNV-specific primers that amplify regions of the L (LdetF/LdetR) and the S (SdetF/SdetR) RNAs (1). Amplification products of the expected 297 and 861 bp size, respectively, were detected in all symptomatic samples while no amplification products were generated from healthy soybean plant tissues grown under greenhouse conditions. Significantly, this is the first documentation and confirmation of the widespread prevalence of SVNV across the state of Michigan in 2012. References: (1) D. L. Smith et al. Plant Dis. http://dx.doi.org/10.1094/PDIS-11-12-1096-PDN . (2) J. Zhou et al. Virus Genes 43:289, 2011.


Mental disorders in patients with noncardiac chest pain.
About 70% of noncardiac chest pain (NCCP) patients have mental disorders as reported from Western countries. The phenomenon of somatization is considered to be aetiologically important in the genesis of NCCP. Though somatization is generally considered as more prevalent among non-Western cultures, systematic studies of mental disorder among NCCP patients are rare from developing countries. Based on treadmill test, 54 male inpatients in a cardiology general ward in India were divided into a group having ischaemic heart disease (IHD) and another group of NCCP. A psychiatric interview was conducted blindly on these subjects with the help of a structured interview schedule, and DSM-III-R diagnosis was made. Sixty-eight percent in the NCCP group and 27% in the IHD group had a mental disorder. The total number of subjects with a mental disorder, including panic disorder and major depression, were significantly more in the NCCP group. Mental disorders appears to be equally common among NCCP patients in developing countries also and detailed psychiatric assessment is warranted in patients with chest pain of non-IHD origin.


Iron overload involved in the enhancement of unloading-induced bone loss by hypomagnetic field.
During deep-space exploration missions, astronauts will be exposed to abnormal space environments including microgravity and hypomagnetic field (HyMF) that is 10,000 times weaker than geomagnetic field (GMF). It is well known that microgravity in space can induce bone loss; however, it is ill-defined whether HyMF involved in this process. Herein, we aimed to investigate the combined effects of HyMF and microgravity on bone loss. A mouse model of hindlimb suspension (HLU) was adopted to simulate microgravity-induced bone loss, that was exposed to a hypomagnetic field of <300 nanotesla (nT) generated by a geomagnetic field-shielding chamber. Besides, a recent study showed that HLU induced bone loss was orchestrated by iron overload. Therefore, the changes of iron content in unloading-induced bone loss under HyMF condition were detected simultaneously. The results showed HyMF exacerbated the loss of bone mineral content (BMC), induced more detrimental effects on microstructure of cancellous bone but not cortical bone and yielded greater negative effects on biomechanical characteristics in mice femur under unloading status. Concomitantly, there was more iron accumulation in serum, liver, spleen and bone in the combined treatment group than in the separate unloading group or HyMF exposure group. These results showed that HyMF promoted additional bone loss in mice femur during mechanical unloading, and the potential mechanism may be involved in inducing iron overload of mice.


Adjunctive Agents for Cervical Preparation in Second Trimester Surgical Abortion.
Late second trimester dilation and evacuation is a challenging subset of surgical abortion. Among the reasons for this is the degree of cervical dilation required to safely extricate fetal parts. Cervical dilation is traditionally achieved by placing multiple sets of osmotic dilators over two or more days prior to the evacuation procedure; however, there is interest in shortening cervical preparation time. The use of adjuvant mifepristone and misoprostol in conjunction with osmotic dilators has been studied for this purpose, and their use demonstrates that adequate cervical dilation can be achieved in less time than with dilators alone. We present a review of the current evidence surrounding adjunctive agents for cervical preparation, and contend that for women presenting for surgical abortion care above 19 weeks gestation, the use of adjunctive mifepristone and/or misoprostol should be strongly considered along with osmotic dilator insertion when cervical preparation in less than 24 h is needed.


Allosteric activation of bacterial response regulators: the role of the cognate histidine kinase beyond phosphorylation.
Response regulators are proteins that undergo transient phosphorylation, connecting specific signals to adaptive responses. Remarkably, the molecular mechanism of response regulator activation remains elusive, largely because of the scarcity of structural data on multidomain response regulators and histidine kinase/response regulator complexes. We now address this question by using a combination of crystallographic data and functional analyses in vitro and in vivo, studying DesR and its cognate sensor kinase DesK, a two-component system that controls membrane fluidity in Bacillus subtilis. We establish that phosphorylation of the receiver domain of DesR is allosterically coupled to two distinct exposed surfaces of the protein, controlling noncanonical dimerization/tetramerization, cooperative activation, and DesK binding. One of these surfaces is critical for both homodimerization- and kinase-triggered allosteric activations. Moreover, DesK induces a phosphorylation-independent activation of DesR in vivo, uncovering a novel and stringent level of specificity among kinases and regulators. Our results support a model that helps to explain how response regulators restrict phosphorylation by small-molecule phosphoryl donors, as well as cross talk with noncognate sensors. The ability to sense and respond to environmental variations is an essential property for cell survival. Two-component systems mediate key signaling pathways that allow bacteria to integrate extra- or intracellular signals. Here we focus on the DesK/DesR system, which acts as a molecular thermometer in B. subtilis, regulating the cell membrane's fluidity. Using a combination of complementary approaches, including determination of the crystal structures of active and inactive forms of the response regulator DesR, we unveil novel molecular mechanisms of DesR's activation switch. In particular, we show that the association of the cognate histidine kinase DesK triggers DesR activation beyond the transfer of the phosphoryl group. On the basis of sequence and structural analyses of other two-component systems, this activation mechanism appears to be used in a wide range of sensory systems, contributing a further level of specificity control among different signaling pathways.


Amiloride enhances the secretion but not the synthesis of renin in renal juxtaglomerular cells.
In this study we have examined a potential role of the sodium/proton exchange system in the regulation of renin secretion. We found that the inhibitors of the Na+/H+ antiport, amiloride (1 mM) and ethylisopropylamiloride (EIPA, 50 microM), led to a 125% increase of renin secretion from cultured mouse juxtaglomerular cells. The stimulatory effect of EIPA on renin secretion was dependent on the extracellular concentrations of sodium and hydrogen ions. While lowering the extracellular pH from 7.3 to 7.0, and lowering [Na+]e from 130 mM to 5 mM had no effect on basal renin release, it markedly attenuated or even blunted the effect of EIPA on renin secretion. The stimulatory effect of forskolin on renin secretion, however, was not altered by decreases of extracellular pH and of sodium. Inhibition of basal renin release was achieved with angiotensin II (1 microM). In the presence of EIPA the inhibitory effect angiotensin II was markedly attenuated. Although effective on renin secretion, neither amiloride nor EIPA exerted a significant effect on the denovo synthesis of renin in cultured mouse JG cells. These findings are compatible with the idea that an amiloride-sensitive transport process, presumably the Na+/H+ exchanger, acts indirectly as an inhibitory signal transduction system for renin secretion from renal juxtaglomerular cells.


Denervation and reinnervation in congenital brachial palsy.
Motor unit number estimation (MUNE) was shown to be useful in assessing the neurophysiological status of 18 subjects with congenital brachial palsy. This was especially so since conventional M-wave measurements may give misleading impressions as to the extent of motor axon regeneration. In most subjects the involvement of sensory nerve fibers indicated that the traumatic lesions included postganglionic segments of the fibers, with or without preganglionic damage. In a minority the lesions were purely preganglionic. Digital sensory nerve involvement was more in a mediolateral direction, consistent with greater damage to the uppermost elements in the brachial plexus. In 5 individuals, MUNE and sensory testing showed that there had been trauma to the supposedly unaffected arm. Discrepancies between sensory and motor results suggested that reinnervation of the biceps brachii muscle was greater than that of the intrinsic muscles of the hand. In one subject examined serially, reinnervation of the hand muscles was detected by 10 months and continued in the hypothenar muscles for the next 6 years.


Understanding non-return after a temporary deferral from giving blood: a qualitative study.
The reasons why deferral from blood donation reduces the likelihood of future return remain unclear. This aim of this study was to investigate possible reasons why deferral has such a dramatic impact on donation patterns. Qualitative methods were used to explore donors' motivations to give blood, their experiences of temporary deferral, and their intentions to return once eligible. Semi-structured interviews were conducted with 23 donors in the two weeks following a temporary deferral due to a low haemoglobin concentration. The Framework approach was used to analyse data and identify themes associated with prompt return, ascertained from Blood Service records. We found that, predominantly, individuals give blood because it represents an easy and convenient way to help others, and provides personal rewards, such as enhancing positive self-concepts and valuable knowledge about health. Deferral disrupts the habit of regular donation, and additionally, introduces an element of practical and emotional hassle to what is generally seen as an undemanding activity. Return after deferral was related to four aspects of a person and their context: an individual's other obligations, especially parenting; whether donation arrangements were facilitated by a range of supports; the presence of a strong "blood donor" identity; and whether deferral left the donor feeling valued and appreciated. Aspects of the deferral process need to be improved to ensure individuals feel valued, and continued attention should be given to the convenience of donation, especially for those with competing obligations.


Gastric juice acidity in upper gastrointestinal diseases.
To search the independent factors determining gastric juice acidity and to investigate the acidity of gastric juices in various benign and malignant upper gastrointestinal diseases. Fasting gastric juice acidity of 165 healthy subjects and 346 patients with esophageal ulcer (n = 21), gastric ulcer (n = 136), duodenal ulcer (n = 100) or gastric cancer (n = 89) were measured and compared. Additionally, gastric specimens were taken from the antrum and body for rapid urease test and histological examination. Multivariate analysis revealed that bile stain of gastric juice, high acute inflammatory score of the corpus, and atrophy of the corpus were independent risk factors for the development of gastric hypoacidity with odds ratios of 3.1 (95% CI: 1.3-7.3), 3.1 (95% CI: 1.2-7.9) and 3.5 (95% CI: 1.3-9.2). Esophageal ulcer and duodenal ulcer patients had a lower pH level (1.9 and 2.1 vs 2.9, both P < 0.05) of gastric juices than healthy subjects. In contrast, gastric ulcer and gastric cancer patients had a higher pH level (3.4 and 6.6 vs 2.9, both P < 0.001) than healthy controls. Hypoacidity existed in 22%, 5%, 29%, 5% and 88% of healthy subjects, esophageal ulcer, gastric ulcer, duodenal ulcer and gastric cancer patients, respectively. Bile reflux, atrophy and dense neutrophil infiltrate of the corpus are three independent factors determining the acidity of gastric juice.


Tetrameric c-di-GMP mediates effective transcription factor dimerization to control Streptomyces development.
The cyclic dinucleotide c-di-GMP is a signaling molecule with diverse functions in cellular physiology. Here, we report that c-di-GMP can assemble into a tetramer that mediates the effective dimerization of a transcription factor, BldD, which controls the progression of multicellular differentiation in sporulating actinomycete bacteria. BldD represses expression of sporulation genes during vegetative growth in a manner that depends on c-di-GMP-mediated dimerization. Structural and biochemical analyses show that tetrameric c-di-GMP links two subunits of BldD through their C-terminal domains, which are otherwise separated by ~10 Å and thus cannot effect dimerization directly. Binding of the c-di-GMP tetramer by BldD is selective and requires a bipartite RXD-X8-RXXD signature. The findings indicate a unique mechanism of protein dimerization and the ability of nucleotide signaling molecules to assume alternative oligomeric states to effect different functions.


Visualization of eukaryotic DNA mismatch repair reveals distinct recognition and repair intermediates.
DNA mismatch repair (MMR) increases replication fidelity by eliminating mispaired bases resulting from replication errors. In Saccharomyces cerevisiae, mispairs are primarily detected by the Msh2-Msh6 complex and corrected following recruitment of the Mlh1-Pms1 complex. Here, we visualized functional fluorescent versions of Msh2-Msh6 and Mlh1-Pms1 in living cells. We found that the Msh2-Msh6 complex is an S phase component of replication centers independent of mispaired bases; this localized pool accounted for 10%-15% of MMR in wild-type cells but was essential for MMR in the absence of Exo1. Unexpectedly, Mlh1-Pms1 formed nuclear foci that, although dependent on Msh2-Msh6 for formation, rarely colocalized with Msh2-Msh6 replication-associated foci. Mlh1-Pms1 foci increased when the number of mispaired bases was increased; in contrast, Msh2-Msh6 foci were unaffected. These findings suggest the presence of replication machinery-coupled and -independent pathways for mispair recognition by Msh2-Msh6, which direct formation of superstoichiometric Mlh1-Pms1 foci that represent sites of active MMR.


[Treatment-resistant psychosis due to interaction between ritonavir and olanzapine: case report and literature review].
Patients with schizophrenia are eight times more likely than healthy individuals to become infected with the human immunodeficiency virus (hiv). Although in vitro studies provide some data on the interaction between antipsychotics and retroviral agents, there is a lack of in vitro data on this subject. We describe the case of a 35-year-old patient who suffered from schizophrenia, polydrug abuse and an hiv infection and who also had treatment-resistant psychosis. An interaction between the antiretroviral drug ritonavir and the antipsychotic olanzapine turned out to be the cause of the treatment resistance. As far as we know, this is the first report of such a clinically relevant interaction. We present a review of the current literature on this type of interaction.


A hexanucleotide element directs microRNA nuclear import.
MicroRNAs (miRNAs) negatively regulate partially complementary target messenger RNAs. Target selection in animals is dictated primarily by sequences at the miRNA 5' end. We demonstrated that despite their small size, specific miRNAs contain additional sequence elements that control their posttranscriptional behavior, including their subcellular localization. We showed that human miR-29b, in contrast to other studied animal miRNAs, is predominantly localized to the nucleus. The distinctive hexanucleotide terminal motif of miR-29b acts as a transferable nuclear localization element that directs nuclear enrichment of miRNAs or small interfering RNAs to which it is attached. Our results indicate that miRNAs sharing common 5' sequences, considered to be largely redundant, might have distinct functions because of the influence of cis-acting regulatory motifs.


CD86 Expression by Monocytes Influences an Immunomodulatory Profile in Asymptomatic Patients with Chronic Chagas Disease.
In the chronic phase of Chagas disease, 60% of the patients develop the asymptomatic form known as indeterminate (IND). The remaining 30% of the patients develop a life-threatening form in which digestive and/or cardiac (CARD) alterations take place. The mechanisms underlying the development of severe forms of Chagas disease remain poorly understood. It is well known that interactions between immune cells such as monocytes and lymphocytes drive immune responses. Further, the co-stimulatory molecules CD80 and CD86 expressed by monocytes and subsets induce lymphocyte activation, thereby triggering cellular immune response. Here, we revealed, for the first time, the functional-phenotypic profile of monocytes subsets in Chagas disease. Using flow cytometry, we evaluated the effect of in vitro stimulation with Trypanosoma cruzi antigens on the expression of the co-stimulatory molecules CD80 and CD86 in different monocyte subsets of patients with IND and CARD clinical forms of Chagas disease. We also assessed the expression of toll-like receptor (TLR)-2, TLR-4, TLR-9, HLA-DR, IL-10, and IL-12 in the monocyte subsets and of CTLA-4 and CD28, ligands of CD80 and CD86, in T lymphocytes. CD86 expression in all monocyte subsets was higher in IND patients when compared with non-infected (NI) individuals. After stimulation with T. cruzi, these patients also showed a higher frequency of CD4+CTLA-4+ T lymphocytes than NI individuals. We found an association between CD80 and CD28, and between CD86 and CTLA-4 expression, with a high frequency of regulatory T (Treg) cells in IND patients. We proposed that CD86 may be involved in immunoregulation by its association with CTLA-4 in asymptomatic patients. CD86 and CTLA-4 interaction may influence Treg activation, and this could represent a new strategy to control inflammation and tissue damage.


Reconstruction of pediatric brachial plexus injuries with nerve grafts and nerve transfers.
To review the demographics and injury patterns in consecutive pediatric patients with traumatic brachial plexus injury presenting to a single center over a 16-year period and to review the outcomes of nerve grafting and nerve transfers for reconstruction of shoulder abduction and elbow flexion in these patients. Forty-five pediatric patients presented for treatment of traumatic Brachial plexus injury from 1996 to 2012. Subgroup analysis of patients who had nerve grafting or nerve transfers for restoration of shoulder abduction and elbow flexion was carried out to compare outcomes of Medical Research Council (MRC) motor grading. The mean age of patients was 13.8 years (range, 3-17 y). Panplexal injuries (62%) and upper plexus injuries (16%) were particularly common. There was a very high proportion of preganglionic injuries (91%). Six of the 10 of patients who underwent intraplexal nerve grafting only for restoration of shoulder abduction achieved grade 3 or better power compared with 42% (5/12) of patients who had nerve transfers. When contralateral C7 was used as a donor for nerve transfer in restoration of shoulder abduction, 1 of the 5 patients achieved grade 3 or better shoulder abduction. All 4 patients who had nerve grafts for restoration of elbow flexion achieved grade 3 or better power, compared with 11 of 12 patients who had nerve transfers. There was no statistical difference in outcome (MRC grade 3 or 4) between patients who had nerve grafts and those who had nerve transfers. This study shows that nerve grafts can result in similar outcomes (MRC grading) to nerve transfers for restoration of shoulder abduction and elbow flexion in traumatic pediatric BPI. The findings of this study do not support the use of contralateral C7 as a donor for nerve transfer in reconstruction of shoulder abduction in this age group. Therapeutic IV.


OpenLabNotes--An Electronic Laboratory Notebook Extension for OpenLabFramework.
Electronic laboratory notebooks (ELNs) are more accessible and reliable than their paper based alternatives and thus find widespread adoption. While a large number of commercial products is available, small- to mid-sized laboratories can often not afford the costs or are concerned about the longevity of the providers. Turning towards free alternatives, however, raises questions about data protection, which are not sufficiently addressed by available solutions. To serve as legal documents, ELNs must prevent scientific fraud through technical means such as digital signatures. It would also be advantageous if an ELN was integrated with a laboratory information management system to allow for a comprehensive documentation of experimental work including the location of samples that were used in a particular experiment. Here, we present OpenLabNotes, which adds state-of-the-art ELN capabilities to OpenLabFramework, a powerful and flexible laboratory information management system. In contrast to comparable solutions, it allows to protect the intellectual property of its users by offering data protection with digital signatures. OpenLabNotes effectively closes the gap between research documentation and sample management, thus making Open-LabFramework more attractive for laboratories that seek to increase productivity through electronic data management.


Afatinib With Pembrolizumab for Treatment of Patients With Locally Advanced/Metastatic Squamous Cell Carcinoma of the Lung: The LUX-Lung IO/KEYNOTE 497 Study Protocol.
Afatinib is a selective, irreversible ErbB family blocker that has shown survival benefit in lung squamous-cell carcinoma (SCC) patients. Pembrolizumab, a humanized immunoglobulin G4 monoclonal antibody to the programmed cell death 1 (PD-1) receptor, has also shown survival benefit in lung SCC. Concurrent inhibition of the PD-1 and epidermal growth factor receptor (EGFR) pathways represents a rational approach to improve responses and delay the onset of treatment resistance in lung SCC. This phase II, open-label, single-arm study (NCT03157089) is designed to assess the efficacy and safety of afatinib in combination with pembrolizumab in patients with stage IIIB/IV lung SCC that has progressed during/after first-line platinum-based chemotherapy. Eligible patients must have ≥1 target lesion (as per Response Evaluation Criteria in Solid Tumors version 1.1) and must have not received previous immune checkpoint inhibitor/EGFR-targeted therapy. The recommended phase II dose (RP2D) and safety profile will be determined during a safety run-in with oral afatinib (starting dose, 40 mg/d) with intravenous pembrolizumab (200 mg every 3 weeks). In the main study, all patients will receive afatinib at the RP2D with pembrolizumab until disease progression, unacceptable toxicity, or for up to 35 cycles. The primary end point is objective response (complete + partial response). Other end points include disease control, duration of objective response, progression-free survival, overall survival, tumor shrinkage, RP2D, and pharmacokinetics. Exploratory biomarker analysis will be performed. This study is being conducted in the United States, Spain, France, South Korea, and Turkey. Enrollment commenced in September 2017, with a target of 50 to 62 patients.


Combining probabilistic forecasts of COVID-19 mortality in the United States.
The COVID-19 pandemic has placed forecasting models at the forefront of health policy making. Predictions of mortality, cases and hospitalisations help governments meet planning and resource allocation challenges. In this paper, we consider the weekly forecasting of the cumulative mortality due to COVID-19 at the national and state level in the U.S. Optimal decision-making requires a forecast of a probability distribution, rather than just a single point forecast. Interval forecasts are also important, as they can support decision making and provide situational awareness. We consider the case where probabilistic forecasts have been provided by multiple forecasting teams, and we combine the forecasts to extract the wisdom of the crowd. We use a dataset that has been made publicly available from the COVID-19 Forecast Hub. A notable feature of the dataset is that the availability of forecasts from participating teams varies greatly across the 40 weeks in our study. We evaluate the accuracy of combining methods that have been previously proposed for interval forecasts and predictions of probability distributions. These include the use of the simple average, the median, and trimming methods. In addition, we propose several new weighted combining methods. Our results show that, although the median was very useful for the early weeks of the pandemic, the simple average was preferable thereafter, and that, as a history of forecast accuracy accumulates, the best results can be produced by a weighted combining method that uses weights that are inversely proportional to the historical accuracy of the individual forecasting teams.


Efficient retrieval of landscape Hessian: forced optimal covariance adaptive learning.
Knowledge of the Hessian matrix at the landscape optimum of a controlled physical observable offers valuable information about the system robustness to control noise. The Hessian can also assist in physical landscape characterization, which is of particular interest in quantum system control experiments. The recently developed landscape theoretical analysis motivated the compilation of an automated method to learn the Hessian matrix about the global optimum without derivative measurements from noisy data. The current study introduces the forced optimal covariance adaptive learning (FOCAL) technique for this purpose. FOCAL relies on the covariance matrix adaptation evolution strategy (CMA-ES) that exploits covariance information amongst the control variables by means of principal component analysis. The FOCAL technique is designed to operate with experimental optimization, generally involving continuous high-dimensional search landscapes (≳30) with large Hessian condition numbers (≳10^{4}). This paper introduces the theoretical foundations of the inverse relationship between the covariance learned by the evolution strategy and the actual Hessian matrix of the landscape. FOCAL is presented and demonstrated to retrieve the Hessian matrix with high fidelity on both model landscapes and quantum control experiments, which are observed to possess nonseparable, nonquadratic search landscapes. The recovered Hessian forms were corroborated by physical knowledge of the systems. The implications of FOCAL extend beyond the investigated studies to potentially cover other physically motivated multivariate landscapes.


Eugen Bleuler's concept of schizophrenia and its relevance to present-day psychiatry.
This paper links the historical perspective with the actual debate on the concept of schizophrenia. By this, two aims shall be accomplished. First, to prove that Eugen Bleuler's (1857-1939) concept of 'schizophrenia' in its central parts was a clear step forward, as compared to previous approaches, especially the notion of 'dementia praecox', proposed and favored by French authors like Bénédict Augustin Morel (1809-1873) and, in Germany, by Emil Kraepelin (1856-1926). Bleuler considerably reduced the epistemological presuppositions of Kraepelin's nosological model and coined the term 'group of schizophrenias', which was markedly broader and, as for prognosis, much less pessimistic. The second aim of this paper is to argue in favor of a continuous reflection upon psychiatry's historical and epistemological basis which is regarded not just as 'l'art pour l'art', but as an indispensable component of psychiatry, clinically and scientifically.


A negative regulatory region in the intracellular domain of the human interferon-alpha receptor.
Interferon-alpha (IFN-alpha)-mediated intracellular signaling is initiated by ligand-induced receptor dimerization, tyrosine phosphorylation of the Tyk2 and Jak1 tyrosine kinases, and subsequent phosphorylation of the Stat1 and Stat2 proteins. The IFN-alpha receptor consists of at least two distinct subunits. One subunit, IFNAR1, has low affinity binding for interferon yet is required for signal transduction. We introduced mutations in the cytoplasmic domain of human IFNAR1 in order to identify residues involved in the mediation of biological responses. We took advantage of the species specificity of the interferon receptors by analyzing human IFN-alpha-induced major histocompatibility complex class I antigen expression in mouse L929 cells stably transfected with mutant human receptors. The membrane proximal 60-amino acids were insufficient to signal a biological response even though within these residues Tyk2 and Stat2 binding sites have been identified. IFN-alpha-induced receptor tyrosine phosphorylation was not critical for signaling because mutation of Tyr residues to Phe did not prevent the biological response to IFN-alpha. The deletion of a 16-amino acid region highly homologous between species created a receptor which signals an enhanced response. Tyrosine dephosphorylation is a component of this enhanced response as mutation of the Tyr residues within this region to Phe resulted in a receptor with increased sensitivity to IFN. The known signaling molecules that interact with IFNAR1 are positive regulators of IFN-alpha function. The presence of this domain in the COOH-terminal region suggests that the receptor may interact with signaling molecules that negatively regulate interferon responses.


Sustaining a regional emerging infectious disease research network: a trust-based approach.
The Asia Partnership on Emerging Infectious Diseases Research (APEIR) was initiated in 2006 to promote regional collaboration in avian influenza research. In 2009, the partnership expanded its scope to include all emerging infectious diseases. APEIR partners include public health and animal researchers, officials and practitioners from Cambodia, China, Lao PDR, Indonesia, Thailand and Vietnam. APEIR has accomplished several major achievements in three key areas of activity: (i) knowledge generation (i.e., through research); (ii) research capacity building (e.g., by developing high-quality research proposals, by planning and conducting joint research projects, by adopting a broader Ecohealth/OneHealth approach); and (iii) policy advocacy (e.g., by disseminating research results to policy makers). This paper describes these achievements, with a focus on the partnership's five major areas of emerging infectious disease research: wild migratory birds, backyard poultry systems, socio-economic impact, policy analysis, and control measures. We highlight two case studies illustrating how the partnership's research results are being used to inform policy. We also highlight lessons learned after five years of working hard to build our partnership and the value added by a multi-country, multi-sectoral, multi-disciplinary research partnership like APEIR.


Akt suppresses DLK for maintaining self-renewal of mouse embryonic stem cells.
Mouse embryonic stem cells (ES cells) can proliferate indefinitely. To identify potential signals involved in suppression of self-renewal, we previously screened a kinase/phosphatase expression library in ES cells, and observed that inhibition of Dual Leucine zipper-bearing Kinase (DLK) increased relative cell numbers. DLK protein was detected in both the pluripotent and differentiated states of mouse ES cells while DLK kinase activity increased upon differentiation. Overexpression of DLK in mouse ES cells displayed reductions in relative cell/colony numbers and Nanog expression, suggesting a suppressive role of DLK in self-renewal. By examining protein sequences of DLK, we identified 2 putative Akt phosphorylation sites at S584 and T659. Blocking PI3K/Akt signaling with LY-294002 enhanced DLK kinase activity dramatically. We found that Akt interacts with and phosphorylates DLK. Mutations of DLK amino acid residues at putative Akt phosphorylation sites (S584A, T659A, or S584A and T659A) diminished the level of DLK phosphorylation. While the mutated DLKs (S584A, T659A, or S584A and T659A) were expressed, a further reduction in cell/colony numbers and Nanog expression appeared in mouse ES cells. In addition, these mutant DLKs (S584A, T659A, or S584A and T659A) exhibited more robust kinase activity and cell death compared to wild type DLK or green fluorescence (GFP) controls. In summary, our results show that DLK functions to suppress self-renewal of mouse ES cells and is restrained by Akt phosphorylation.


T cell recognition of endogenous IgG2a expressed in B lymphoma cells.
We recently characterized a panel of C57BL/6J T cell clones specific for IgG2a of the a allotype in association with I-Ab. Several of the clones gave surprisingly strong responses in the presence of normal spleen cells from several H-2b strains, including C3H.SW, A.BY, D1.LP and BALB.B, without the addition of exogenous antigen. Experiments using Igh-congenic mouse strains demonstrated that this response was directed towards shared allotypic determinants expressed on endogenously synthesized IgG2a molecules in various strains. Here we present evidence that the cell(s) responsible for this stimulation are conventional low density splenic accessory cells. Presentation of endogenously synthesized IgG2a by this population of dendritic cells, macrophages, and/or activated B cells was chloroquine sensitive. Thus, we conclude that this response is probably directed towards secreted IgG2a molecules that are internalized, processed and re-expressed at the cell surface in association with class II molecules. We also tested the ability of T cell clone B61-34 to respond in the presence of the B lymphoma cell line M12.C3.A2. A strong response was again observed in the absence of exogenous antigen. Northern gel analysis of M12.C3.A2 messenger RNA provided evidence that these cells synthesize IgG2a in both a secreted and a membrane form. The response directed towards endogenous IgG2a expressed in M12.C3.A2 lymphoma cells is chloroquine resistant and considerably more efficient than that stimulated by IgG2a added as exogenous soluble antigen. These results demonstrate for the first time that B cells have the capacity to present antigenic determinants expressed on endogenously synthesized immunoglobulins to class II-restricted T cells. The implications of these findings with respect to network models of immune regulation are discussed.


Current Therapeutic Molecules and Targets in Neurodegenerative Diseases Based on in silico Drug Design.
As the number of elderly persons increases, neurodegenerative diseases are becoming ubiquitous. There is currently a great need for knowledge concerning management of oldage neurodegenerative diseases; the most important of which are: Alzheimer's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis, and Huntington's disease. To summarize the potential of computationally predicted molecules and targets against neurodegenerative diseases. Review of literature published since 1997 against neurodegenerative diseases, utilizing as keywords: in silico, Alzheimer's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis ALS, and Huntington's disease was conducted. Due to the costs associated with experimentation and current ethical law, performing experiments directly on living organisms has become much more difficult. In this scenario, in silico techniques have been successful and have become powerful tools in the search to cure disease. Researchers use the Computer Aided Drug Design pipeline which: 1) generates 3- dimensional structures of target proteins through homology modeling 2) achieves stabilization through molecular dynamics simulation, and 3) exploits molecular docking through large compound libraries. Next generation sequencing is continually producing enormous amounts of raw sequence data while neuroimaging is producing a multitude of raw image data. To solve such pressing problems, these new tools and algorithms are required. This review elaborates precise in silico tools and techniques for drug targets, active molecules, and molecular docking studies, together with future prospects and challenges concerning possible breakthroughs in Alzheimer's, Parkinson's, Amyotrophic Lateral Sclerosis, and Huntington's disease.


Molecular detection of acute lymphoblastic leukaemia in boys with testicular relapse.
To determine the role of polymerase chain reaction (PCR) based minimal residual disease (MRD) detection of leukaemia specific DNA in testicular relapse in childhood acute lymphoblastic leukaemia. DNA was obtained from archival testicular and bone marrow samples from boys with acute lymphoblastic leukaemia who relapsed in the testes. Overlapping DJH clone specific primers derived from clonal immunoglobulin heavy chain (IgH) gene rearrangement in each case were used to analyse testicular or bone marrow DNA. Histologically normal end of treatment testicular biopsies in the five patients in longterm remission were all MRD negative, but MRD positive in three of six boys with subsequent testicular relapse. Histologically normal bone marrow samples taken at the end of treatment were MRD negative in five of seven cases, but MRD positive in all cases at the time of isolated testicular relapse. Three boys with unilateral testicular relapse underwent unilateral orchidectomy, rather than bilateral testicular irradiation, as part of their treatment. Two of these boys were MRD positive in the histologically uninvolved testes, and both had subsequent relapses either in the testes or the bone marrow, while the MRD negative patient has not had a testicular relapse. The presence of MRD in testicular tissue can be assayed with a PCR based method to detect clone specific antigen receptor gene rearrangements. In this setting, PCR is more sensitive than conventional testicular histology for predicting clinical outcomes. MRD assays might be useful in the management of boys at the time of isolated testicular relapse, to confirm the presence of unilateral testicular disease.


Depression and impulse control disorders in Parkinson's disease: two sides of the same coin?
Depression and impulse control disorders (ICD) are two common neuropsychiatric features in Parkinson's disease (PD). Studies have revealed that both phenomena are associated with aberrations in ventral striatal dopamine signaling and concomitant dysfunction of the reward-related (limbic) cortico-striatal-thalamocortical (CSTC) circuit. Depression in PD seems associated with decreased activity in the limbic CSTC circuit, whereas ICD seem associated with increased limbic CSTC circuit activity, usually after commencing dopamine replacement therapy (DRT). Not all DRT using PD patients, however, develop symptoms of ICD, suggesting an additional underlying neurobiological susceptibility. Furthermore, the symptoms of depression and ICD frequently coincide even though they are related to seemingly contrasting limbic CSTC circuit activation states. The aim of this review is to provide an overview of the currently available literature on the neurobiology of PD-related depression and ICD and discusses possible susceptibility factors. Finally, we propose a neurobiological model that identifies ventral striatal dopaminergic denervation as a common underlying neurobiological substrate of depression and ICD and subsequent dysfunction of reward and motivation-related brain areas.


L-ficolin binds to the glycoproteins hemagglutinin and neuraminidase and inhibits influenza A virus infection both in vitro and in vivo.
L-ficolin, one of the complement lectins found in human serum, is a novel pattern recognition molecule that can specifically bind to microbial carbohydrates, thereby activating the lectin complement pathway and mounting a protective innate immune response. However, little is known about the role of L-ficolin during viral infections in vivo. In the present study, we used a mouse model of influenza A virus infection to demonstrate that the administration of exogenous L-ficolin or ficolin A (FCNA - an L-ficolin-like molecule in the mouse) is protective against the virus. Furthermore, FCNA-null mice have a greatly increased susceptibility to infection with the influenza A virus. Moreover, we found recombinant human L-ficolin inhibited influenza A virus entry into Madin-Darby canine kidney cells. More importantly, L-ficolin can recognize and bind hemagglutinin (HA) and neuraminidase (NA) glycoproteins and different subtypes of influenza A virus, and these interactions can be competitively inhibited by N-acetyl-D-glucosamine. In addition, the binding of L-ficolin and FCNA may lead to the activation of the lectin complement pathway. To our knowledge, this is the first report demonstrating that L-ficolin can block influenza virus infections both in vitro and in vivo using FCNA-knockout mice, possibly by interacting with the carbohydrates of HA and NA. Therefore, these data may provide new immunotherapeutic strategies based on the innate immune molecule L-ficolin against the influenza A virus.


Self-Directed Learning: A Cognitive and Computational Perspective.
A widely advocated idea in education is that people learn better when the flow of experience is under their control (i.e., learning is self-directed). However, the reasons why volitional control might result in superior acquisition and the limits to such advantages remain poorly understood. In this article, we review the issue from both a cognitive and computational perspective. On the cognitive side, self-directed learning allows individuals to focus effort on useful information they do not yet possess, can expose information that is inaccessible via passive observation, and may enhance the encoding and retention of materials. On the computational side, the development of efficient "active learning" algorithms that can select their own training data is an emerging research topic in machine learning. This review argues that recent advances in these related fields may offer a fresh theoretical perspective on how people gather information to support their own learning.


Knowledge on Antibiotic Use, Self-Reported Adherence to Antibiotic Intake, and Knowledge on Multi-Drug Resistant Pathogens - Results of a Population-Based Survey in Lower Saxony, Germany.
Assessment of public awareness on antibiotic use and resistance can identify key issues for campaigns addressing these problems. Our aim was to assess the knowledge, attitudes, and practice (KAP) related to antibiotic use and multi-drug resistant (MDR) pathogens in a general population in Germany. We conducted a KAP survey on antibiotics and on MDR pathogens using an online panel recruited from the general population, which was established using stratified random sampling from the population registry in four districts in Lower Saxony, Germany. In the 12 months preceding the survey, 32.3% of the participants had received at least one prescription for antibiotics, 95.7% reported to follow the recommendations of prescribers, and 10.3% reported to stop taking antibiotics as soon as they feel better. Up to 94.9% of the participants had heard of MDR pathogens, 42.7% reported to know somebody who had been tested positive for it, 0.8% had an infection with it, and 37.2% were worried of contracting it. In case of contact with a carrier of MDR pathogens, over 90% would increase hand hygiene and 0.8% would avoid the carrier completely. Participants considered health care workers (75.1%) and everybody in society (87.8%) to be responsible for combating the spread of MDR pathogens. There is a high reported exposure to antibiotics and awareness of the problem of MDR pathogens. Despite personal worries, most of the participants indicated a reasonable, non-stigmatizing behavior toward carriers of MDR pathogens, and that every individual was responsible to avoid their spread.


Occurrence of leu+ revertants under starvation cultures in Escherichia coli is growth-dependent.
Many investigations have reported that advantageous mutations occurred more frequently under selective conditions than those under non-selective conditions. This phenomenon is referred to as adaptive mutation. Their characteristics are that adaptive mutations are directed and growth-independent. The idea of directed adaptive mutation had been objected by some reports, however, the idea of growth-independent adaptive mutation has been held till today. In this paper, we have observed that under leucine starvation conditions, leu+ revertants accumulated as a function of time; leu- to leu+ reverse mutation rates and frequencies were higher than those under non starvation conditions; and no divided cells could be monitored by the penicillin method. These results were similar to the time-dependent manner of adaptive mutation from previous reports. However, leucine concentration determinate experiments revealed that certain traces of leucine, which leaked from the E. coli cells, was almost always present in the culture. More numbers of leu+ revertants appeared when the similar cultures were dropped in small areas on the selective plates than when spread on the whole selective plates. These results have shown that mutations under leucine starving conditions are growth-dependent. Fluctuation analysis of leu+ revertants indicated that leu-leu+ mutation occurred spontaneously and randomly. In addition, the spectra of leuB gene in the revertants proved that mutations under selective conditions were not specific or directed. The above investigations led to the conclusion (1) that the occurrence of leu+ mutations under starvation conditions was growth-dependent. The occurrence mutations was also similar to that under non-starvation conditions (2). Under starvation conditions the mutation rates were higher, and was not constant during the long process.


Inhomogeneity in response to air pollution in European children (PEACE project).
The PEACE study is a multicentre panel study of the acute effects of particles with a 50% cut off aerodynamic diameter of 10 microns (PM10), black smoke (BS), sulphur dioxide (SO2), and nitrogen dioxide (NO2) on respiratory health of children with chronic respiratory symptoms. In the complete panels no consistent association between air pollution and respiratory health was found. The study evaluated whether potentially more sensitive subgroups in the panels did show effects of air pollution. To evaluate heterogeneity in response to air pollution, effect estimates of air pollution on peak expiratory flow (PEF) and respiratory symptoms were calculated in subgroups based on presence of chronic respiratory symptoms, use of respiratory medication, atopy, sex, and baseline lung function. The association between PEF and air pollution was positive in asthmatic children who used respiratory medication whereas the associations tended to be negative in children who did not use respiratory medication selected only on cough. No consistent association was found among asthmatic children who did not use medication. The association between daily prevalence of symptoms and concentrations of air pollution was not different between these subgroups. None of the predefined potentially more sensitive subgroups showed a consistent association between air pollution, PEF, and respiratory symptoms.


A dietary supplement abolishes age-related cognitive decline in transgenic mice expressing elevated free radical processes.
We previously found that transgenic mice overexpressing growth hormone (TGM) have elevated and progressively increasing free radical processes in brain that strongly correlates with reduced survivorship. Young mature TGM, however, displayed vastly enhanced learning of an eight-choice cued maze and qualitatively different learning curves than normal controls. Here we document the age-related patterns in learning ability of TGM and normal mice. Learning appeared inferior in both genotypes of very young mice but TGM were confirmed to be superior to normal mice upon maturity. Older TGM, however, showed rapid age-related loss of their exceptional learning, whereas normal mice at 1 year of age showed little change. The cognitive decline of TGM was abolished by a complex "anti-aging" dietary supplement formulated to promote membrane and mitochondrial integrity, increase insulin sensitivity, reduce reactive oxygen and nitrogen species, and ameliorate inflammation. Results are discussed in the context of reactive oxygen and nitrogen species, long-term potentiation, learning, aging and neuropathology, based on known impacts of the growth hormone axis on the brain, and characteristics of TGM.


Current Concepts in the Epidemiology, Diagnosis, and Management of Histoplasmosis Syndromes.
Histoplasmosis is a global disease endemic to regions of all six inhabited continents. The areas of highest endemicity lie within the Mississippi and Ohio River Valleys of North America and parts of Central and South America. As a result of climate change and anthropogenic land utilization, the conditions suitable for Histoplasma capsulatum are changing, leading to a corresponding change in epidemiology. The clinical manifestations of histoplasmosis are protean, variably resembling other common conditions such as community-acquired pneumonia, tuberculosis, sarcoidosis, Crohn's disease, or malignancy. Making a successful diagnosis is contingent on a thorough understanding of epidemiology, common clinical presentations, and best testing practices for histoplasmosis. While most subclinical or self-limited diseases do not require treatment in immunocompetent patients, all immunocompromised patients and those with progressive disseminated disease or chronic pulmonary disease should be treated. Liposomal amphotericin B is the preferred agent for severe or disseminated disease, while itraconazole is adequate for milder cases and "step-down" therapy following response to amphotericin B. In this review, we discuss the current evidence-based approaches to the epidemiology, diagnosis, and management of histoplasmosis.


Mechanisms of Resistance to Folate Pathway Inhibitors in Burkholderia pseudomallei: Deviation from the Norm.
The trimethoprim and sulfamethoxazole combination, co-trimoxazole, plays a vital role in the treatment of Burkholderia pseudomallei infections. Previous studies demonstrated that the B. pseudomallei BpeEF-OprC efflux pump confers widespread trimethoprim resistance in clinical and environmental isolates, but this is not accompanied by significant resistance to co-trimoxazole. Using the excluded select-agent strain B. pseudomallei Bp82, we now show that in vitro acquired trimethoprim versus co-trimoxazole resistance is mainly mediated by constitutive BpeEF-OprC expression due to bpeT mutations or by BpeEF-OprC overexpression due to bpeS mutations. Mutations in bpeT affect the carboxy-terminal effector-binding domain of the BpeT LysR-type activator protein. Trimethoprim resistance can also be mediated by dihydrofolate reductase (FolA) target mutations, but this occurs rarely unless BpeEF-OprC is absent. BpeS is a transcriptional regulator that is 62% identical to BpeT. Mutations affecting the BpeS DNA-binding or carboxy-terminal effector-binding domains result in constitutive BpeEF-OprC overexpression, leading to trimethoprim and sulfamethoxazole efflux and thus to co-trimoxazole resistance. The majority of laboratory-selected co-trimoxazole-resistant mutants often also contain mutations in folM, encoding a pterin reductase. Genetic analyses of these mutants established that both bpeS mutations and folM mutations contribute to co-trimoxazole resistance, although the exact role of folM remains to be determined. Mutations affecting bpeT, bpeS, and folM are common in co-trimoxazole-resistant clinical isolates, indicating that mutations affecting these genes are clinically significant. Co-trimoxazole resistance in B. pseudomallei is a complex phenomenon, which may explain why resistance to this drug is rare in this bacterium.IMPORTANCEBurkholderia pseudomallei causes melioidosis, a tropical disease that is difficult to treat. The bacterium's resistance to antibiotics limits therapeutic options. The paucity of orally available drugs further complicates therapy. The oral drug of choice is co-trimoxazole, a combination of trimethoprim and sulfamethoxazole. These antibiotics target two distinct enzymes, FolA (dihydrofolate reductase) and FolP (dihydropteroate synthase), in the bacterial tetrahydrofolate biosynthetic pathway. Although co-trimoxazole resistance is minimized due to two-target inhibition, bacterial resistance due to folA and folP mutations does occur. Co-trimoxazole resistance in B. pseudomallei is rare and has not yet been studied. Co-trimoxazole resistance in this bacterium employs a novel strategy involving differential regulation of BpeEF-OprC efflux pump expression that determines the drug resistance profile. Contributing are mutations affecting folA, but not folP, and folM, a folate pathway-associated gene whose function is not yet well understood and which has not been previously implicated in folate inhibitor resistance in clinical isolates.


Ethics, economics, and public financing of health care.
There is a wide variety of ethical arguments for public financing of health care that share a common structure built on a series of four logically related propositions regarding: (1) the ultimate purpose of a human life or human society; (2) the role of health and its distribution in society in advancing this ultimate purpose; (3) the role of access to or utilisation of health care in maintaining or improving the desired level and distribution of health among members of society, and (4) the role of public financing in ensuring the ethically justified access to and utilisation of health care by members of society. This paper argues that economics has much to contribute to the development of the ethical foundations for publicly financed health care. It focuses in particular on recent economic work to clarify the concepts of access and need and their role in analyses of the just distribution of health care resources, and on the importance of economic analysis of health care and health care insurance markets in demonstrating why public financing is necessary to achieve broad access to and utilisation of health care services.


Parental Smoking Cessation: Impacting Children's Tobacco Smoke Exposure in the Home.
There is no safe or risk-free level of tobacco use or tobacco smoke exposure. In this randomized controlled trial, we tested a tobacco control intervention in families and specifically evaluated a tailored cessation intervention for the parents and/or caregivers (Ps/Cs) who were smokers while their children were simultaneously enrolled in tobacco prevention. Ps/Cs and children were recruited from 14 elementary schools across rural and urban settings. Approximately one-fourth (24.3%; n = 110) of the total Ps/Cs enrolled in the randomized controlled trial (n = 453) were smokers, predominantly women (80.9%), with a mean age of 37.7 years. (SD 12.2); 62.7% were African American, 44% had less than a high school education, and 58% earned <$20 000 annually. P/C smokers were offered a tailored cessation intervention in years 1 and 2. Self-report smoking status and saliva cotinine were obtained at baseline, the end of treatment (EOT) and/or year 2, and in the year 4 follow-up. Ps/Cs in the intervention group showed a larger increase in self-reported smoking abstinence over time (EOT: 6.5% [SE = 5.7%]; year 4: 40.6% [SE = 5.7%]) than the control group (EOT: 0.0% [SE = 6.5%]; year 4: 13.2% [SE = 6.4%]; F = 4.82; P = .0306). For cotinine, the intervention group showed a decrease from baseline (239.9 [SE = 1.3]) to EOT 99.3 [SE = 1.4]) and then maintenance through year 4 (109.6 [SE = 1.4]), whereas the control group showed increases from baseline (221.1 [SE = 1.4]) to EOT (239.0 [SE = 1.4]) to year 4 (325.8 [SE = 14]; F = 5.72; P = .0039). This study provides evidence that tailored cessation offered to Ps/Cs in their children's schools during their children's enrollment in tobacco prevention may contribute to more robust success in P/C cessation and a reduction of tobacco smoke exposure in children.


Probing DNA - Transcription Factor Interactions Using Single-Molecule Fluorescence Detection in Nanofluidic Devices.
Single-molecule fluorescence detection offers powerful ways to study biomolecules and their complex interactions. Here, nanofluidic devices and camera-based, single-molecule Förster resonance energy transfer (smFRET) detection are combined to study the interactions between plant transcription factors of the auxin response factor (ARF) family and DNA oligonucleotides that contain target DNA response elements. In particular, it is shown that the binding of the unlabeled ARF DNA binding domain (ARF-DBD) to donor and acceptor labeled DNA oligonucleotides can be detected by changes in the FRET efficiency and changes in the diffusion coefficient of the DNA. In addition, this data on fluorescently labeled ARF-DBDs suggest that, at nanomolar concentrations, ARF-DBDs are exclusively present as monomers. In general, the fluidic framework of freely diffusing molecules minimizes potential surface-induced artifacts, enables high-throughput measurements, and proved to be instrumental in shedding more light on the interactions between ARF-DBDs monomers and between ARF-DBDs and their DNA response element.