One-year repeated cycles of cognitive training (CT) for Alzheimer's disease.
Recent research suggests that a combination of both pharmacological and psychosocial treatments targeting cognitive functions improves cognition in patients with Alzheimer's disease (AD). The present study evaluated the effectiveness of a 1-year cognitive training (CT) by comparing the cognitive performance of 16 patients with AD treated with CT and cholinesterase inhibitors (ChEIs) (experimental group) with the performance of 16 patients treated with a non-specific cognitive treatment and ChEIs (control group). This study was a single-blind randomized controlled trial. The patients in the experimental group received pharmacological treatment and repeated cycles of CT for 1 year, whereas the control group received pharmacological treatment and repeated cycles of non-specific cognitive exercises. The patients in the two groups were administered a variety of neuropsychological tests measuring several cognitive functions (i.e. memory, language, reasoning, executive function, working memory and apraxia), activities of daily living, and depression. After 1 year of training, the experimental group scored significantly higher on the Mini Mental State Examination, the Milan Overall Dementia Assessment battery and in other five neuropsychological tests, compared to the control group. Present results suggest that repeated cycles of CT in patients with AD treated with ChEIs are associated with benefits in several areas of cognitive function.


Hyphal chemotropism in fungal pathogenicity.
The ability to grow as filamentous hyphae defines the lifestyle of fungi. Hyphae are exposed to a variety of chemical stimuli such as nutrients or signal molecules from mating partners and host organisms. How fungi sense and process this chemical information to steer hyphal growth is poorly understood. Saccharomyces cerevisiae and Neurospora crassa have served as genetic models for the identification of cellular components functioning in chemotropism. A recent study in the pathogen Fusarium oxysporum revealed distinct MAPK pathways governing hyphal growth towards nutrient sources and sex pheromones or plant signals, suggesting an unanticipated complexity of chemosensing during fungus-host interactions.


Citizen participation in patient prioritization policy decisions: an empirical and experimental study on patients' characteristics.
Health systems worldwide are grappling with the need to control costs to maintain system viability. With the combination of worsening economic conditions, an aging population and reductions in tax revenues, the pressures to make structural changes are expected to continue growing. Common cost control mechanisms, e.g. curtailment of patient access and treatment prioritization, are likely to be adversely viewed by citizens. It seems therefore wise to include them in the decision making processes that lead up to policy changes. In the context of a multilevel iterative mixed-method design a quantitative survey representative of the German population (N = 2031) was conducted to probe the acceptance of priority setting in medicine and to explore the practicability of direct public involvement. Here we focus on preferences for patients' characteristics (medical aspects, lifestyle and socio-economic status) as possible criteria for prioritizing medical services. A questionnaire with closed response options was fielded to gain insight into attitudes toward broad prioritization criteria of patient groups. Furthermore, a discrete choice experiment was used as a rigorous approach to investigate citizens' preferences toward specific criteria level in context of other criteria. Both the questionnaire and the discrete choice experiment were performed with the same sample. The citizens' own health and social situation are included as explanatory variables. Data were evaluated using corresponding analysis, contingency analysis, logistic regression and a multinomial exploded logit model. The results show that some medical criteria are highly accepted for prioritizing patients whereas socio-economic criteria are rejected.


Influencing factors in the occurrence of injured coliforms in the drinking water distribution system in the city of La Plata, Argentina.
The objective of this study was to evaluate the influencing factors in the occurrence of coliforms in the drinking water in La Plata (Argentina) from June 1999 to June 2001. A total of 180 samples were collected from Rio de La Plata (102 samples) and Puelche Aquifer (78 samples); 45 samples were collected for each of the four seasons. The membrane filter procedure was used for isolating bacteria, and each sample was tested for chlorine and pH. The highest percentage of samples positive for coliforms in the two tested media was obtained in summer while the highest percentage of negative samples was obtained in winter. No Escherichia coli was isolated. The percentage of injured coliforms fluctuated between 70 and 100%. The most frequently isolated bacteria was Enterobacter cloacae in summer, Enterobacter agglomerans in autumn and Klebsiella oxytoca in winter and spring. Significant correlations were observed between coliforms and the distance from the initial treatment point, and with the level of free chlorine. We conclude that drinking water contamination in La Plata occurs in the distribution system due to increased temperatures and reduced disinfectant levels, which result in bacterial regrowth.


The interplay of histone modifications - writers that read.
Histones are subject to a vast array of posttranslational modifications including acetylation, methylation, phosphorylation, and ubiquitylation. The writers of these modifications play important roles in normal development and their mutation or misregulation is linked with both genetic disorders and various cancers. Readers of these marks contain protein domains that allow their recruitment to chromatin. Interestingly, writers often contain domains which can read chromatin marks, allowing the reinforcement of modifications through a positive feedback loop or inhibition of their activity by other modifications. We discuss how such positive reinforcement can result in chromatin states that are robust and can be epigenetically maintained through cell division. We describe the implications of these regulatory systems in relation to modifications including H3K4me3, H3K79me3, and H3K36me3 that are associated with active genes and H3K27me3 and H3K9me3 that have been linked to transcriptional repression. We also review the crosstalk between active and repressive modifications, illustrated by the interplay between the Polycomb and Trithorax histone-modifying proteins, and discuss how this may be important in defining gene expression states during development.


Chromosomes with one, two, three, and four fetal globin genes: molecular and hematologic analysis.
Analysis of DNA from 852 Island Melanesians has revealed a high frequency of single- and triple-gamma-globin genes in this population. Homozygotes for triple- and single-gamma genes have normal hematologic findings, normal hemoglobin F (HbF) levels, and when there is coexisting alpha thalassemia, appropriate levels of Bart's Hb (gamma 4) at birth. In addition, we have identified an individual with a quadruple-gamma gene chromosome who also has a normal HbF level. All single-gamma genes were A gamma, all triple-gamma genes G gamma G gamma A gamma, and the quadruple-gamma gene G gamma G gamma G gamma A gamma. Analysis of G gamma:A gamma ratios in cord bloods and HbF levels in adults showed that these additional gamma genes are expressed and are down regulated appropriately by the fetal to adult Hb switch. Analysis of the restriction enzyme haplotypes of these various chromosomes indicates that intrachromosomal cross-overs are more likely to have produced these variants than interchromosomal recombination events.


Interventions for congenital talipes equinovarus (clubfoot).
Congenital talipes equinovarus (CTEV), which is also known as clubfoot, is a common congenital orthopaedic condition. It is characterised by an excessively turned in foot (equinovarus) and high medial longitudinal arch (cavus). If left untreated it can result in long-term disability, deformity and pain. Interventions can be conservative (such as splinting or stretching) or surgical. To evaluate the effectiveness of interventions for CTEV. We searched CENTRAL (2011, Issue 2), NHSEED (2011, Issue 2), MEDLINE (January 1966 to April 2011), EMBASE (January 1980 to April 2011), CINAHL Plus (January 1937 to April 2011), AMED (1985 to April 2011) and the Physiotherapy Evidence Database (PEDro to April 2011). We checked the references of included studies. Randomised and quasi-randomised controlled trials evaluating interventions for CTEV. Participants were people of all ages with CTEV of either one or both feet. Two authors independently assessed risk of bias in included trials and extracted the data. We contacted authors of included trials for missing information. We collected adverse event information from trials when it was available. We identified 13 trials in which there were 507 participants. The use of different outcome measures prevented pooling of data for meta-analysis even when interventions and participants were comparable. All trials displayed bias in four or more areas. One trial reported on the primary outcome of function, though raw data were not available to be analysed. We were able to analyse data on foot alignment (Pirani score), a secondary outcome, from three trials. The Pirani score is scored from zero to six, in which higher is worse. Two of the trials involved participants at initial presentation. One of them reported that the Ponseti technique significantly improved foot alignment compared to the Kite technique. After 10 weeks of serial casting, the average total Pirani score of the Ponseti group was 1.15 (95% confidence interval 0.98 to 1.32) lower than that of the Kite group. The second trial found the Ponseti technique to be superior to a traditional technique, with average total Pirani scores of the Ponseti participants 1.50 lower (95% confidence interval 0.72 to 2.28) after serial casting and Achilles tenotomy. A trial in which the type of presentation was not reported found no difference between an accelerated Ponseti or standard Ponseti treatment. At the end of serial casting, the average total Pirani scores in the standard group were 0.31 lower (95% confidence interval -0.40 to 1.02) than the accelerated group. Adverse events were not compared in the trial. There is a lack of evidence for different plaster casting products or the addition of botulinum toxin A during the Ponseti technique. There is also a lack of evidence for different types of major foot surgery for CTEV, continuous passive motion treatment following major foot surgery, or treatment of relapsed or neglected cases of CTEV. Most trials did not report on adverse events. In trials evaluating serial casting techniques, adverse events included cast slippage (needing replacement), plaster sores (pressure areas) and skin irritation. Adverse events following surgical procedures included infection and the need for skin grafting. From the limited evidence available, the Ponseti technique may produce better short-term outcomes compared to the Kite technique. An accelerated Ponseti technique may be as effective as a standard technique. We could draw no conclusions from other included trials because of the limited use of validated outcome measures and lack of available raw data. Future randomised controlled trials should address these issues.


Bone mass measured by photonabsorptiometry and radiogrammetry in Wistar rats. Age and sex variations.
Using radiogrammetric and photonabsorptiometric methods, we determined changes in bone mass in Wistar rats as a function of age and sex. In the four age groups studied (10-, 18-, 30-, and 78-week-old rats), males displayed greater bone mass than females. With increasing age, bone mass also increases up to the 78th week. However, the rate of this increase was slower in the 78-week age group. Whereas the length and periosteal diameter of the bones studied were increased in old age groups, the endosteal diameter decreased in females and increased in males. The results show that, unlike human bone, rat bone mass persistently increases with aging, and female rats continue to gain bone at the endosteal site. Furthermore, these findings demonstrate that radiogrammetric morphometry and photonabsorptiometry may be used as an experimental tool to investigate bone mass changes in small animals.


Benefits of venoactive drug therapy in surgical or endovenous treatment for varicose veins: a systematic review.
Lower limb varicose veins can be treated effectively with endovenous ablation procedures or with minimally invasive open techniques. However, these procedures may cause pain and discomfort, and surgical stripping can be associated with long recovery times. We investigated whether venoactive drugs (VAD) used to treat chronic venous disease (CVD), provide benefits to patients recovering from a surgical or endovenous varicose vein procedure. We conducted a systematic review of the literature to identify clinical trials investigating VAD therapy before, during, or after a surgical, endovenous, or sclerotherapy procedure for varicose veins. Records retrieved from the PubMed and Embase databases were screened for relevant studies. Full-length articles in English were obtained for analysis. We identified five clinical trials investigating the effects of VAD on recovery after surgery, endovenous ablation, or sclerotherapy. Studies were conducted in Russia, Italy, and Czech Republic and varied considerably in design, but all were unblinded open-label studies. All studies reported the use of micronized purified flavonoid fraction (MPFF); in one study, sulodexide was also used. Three studies reported significantly less post-procedural pain with MPFF treatment, with one reporting no significant effect. Two studies reported significant reductions in post-procedural bleeding with MPFF treatment. Three out of four studies reported greater symptomatic improvement with MPFF treatment. Appropriate treatment with MPFF may help reduce post-procedural pain, hemorrhage, and CVD-specific symptoms. While adjunct VAD treatment seems promising, high-quality placebo-controlled studies are needed to unequivocally demonstrate its benefits.


The job of 'ethics committees'.
What should authorities establish as the job of ethics committees and review boards? Two answers are: (1) review of proposals for consistency with the duly established and applicable code and (2) review of proposals for ethical acceptability. The present paper argues that these two jobs come apart in principle and in practice. On grounds of practicality, publicity and separation of powers, it argues that the relevant authorities do better to establish code-consistency review and not ethics-consistency review. It also rebuts bad code and independence arguments for the opposite view. It then argues that authorities at present variously specify both code-consistency and ethics-consistency jobs, but most are also unclear on this issue. The paper then argues that they should reform the job of review boards and ethics committees, by clearly establishing code-consistency review and disestablishing ethics-consistency review, and through related reform of the basic orientation, focus, name, and expertise profile of these bodies and their actions.


Assessment of fatigue in intraocular surgery: analysis using a virtual reality simulator.
To determine the effect of fatigue on intraocular surgical performance using a virtual reality simulator. Seven experienced ophthalmic surgeons were recruited. The first set of data collection was immediately before a live theatre session. Each surgeon received a standardized orientation to a virtual reality cataract surgery simulator (Eyesi©, VRmagic, Mannheim, Germany). All surgeons then completed ten attempts on level-four forceps module. The parameters recorded were total score, total time, total time score, corneal injury score, lens injury score, odometer score, and operating without red reflex score. To reduce the effect of the learning curve, each surgeon had a "plateau" score calculated for every parameter, which was the average of their final four attempts. The surgeons then returned immediately after their scheduled theatre lists to complete a further ten attempts on the same module and similar parameters were recorded. Following routine theatre lists, simulator parameters slightly improved, with only the total score (97.28 to 98.57, p = 0.028) and total time (44 s to 35 s, p = 0.033) being statistically significant. The mean theatre list operating time was 197 min (SD ± 23.60). No detrimental effect of fatigue was demonstrated following a routine operating list. The study provides a template for further assessments of fatigue in high-volume cataract surgery lists.


Augmenting Instructional Animations with a Body Analogy to Help Children Learn about Physical Systems.
We investigated whether augmenting instructional animations with a body analogy (BA) would improve 10- to 13-year-old children's learning about class-1 levers. Children with a lower level of general math skill who learned with an instructional animation that provided a BA of the physical system, showed higher accuracy on a lever problem-solving reaction time task than children studying the instructional animation without this BA. Additionally, learning with a BA led to a higher speed-accuracy trade-off during the transfer task for children with a lower math skill, which provided additional evidence that especially this group is likely to be affected by learning with a BA. However, overall accuracy and solving speed on the transfer task was not affected by learning with or without this BA. These results suggest that providing children with a BA during animation study provides a stepping-stone for understanding mechanical principles of a physical system, which may prove useful for instructional designers. Yet, because the BA does not seem effective for all children, nor for all tasks, the degree of effectiveness of body analogies should be studied further. Future research, we conclude, should be more sensitive to the necessary degree of analogous mapping between the body and physical systems, and whether this mapping is effective for reasoning about more complex instantiations of such physical systems.


Use of High-Resolution Pressure Nephelometry To Measure Gas Vesicle Collapse as a Means of Determining Growth and Turgor Changes in Planktonic Cyanobacteria.
Previous work has demonstrated that the physical properties of intracellular bacterial gas vesicles (GVs) can be analyzed in vivo using pressure nephelometry. In analyzing the buoyant state of GV-containing cyanobacteria, hydrostatic pressure within a sample cell is increased in a stepwise manner, where the concomitant collapse of GVs due to pressure and the resultant decrease in suspended cells are detected by changes in nephelometric scattering. As the relative pressure at which GVs collapse is a function of turgor pressure and cellular osmotic gradients, pressure nephelometry is a powerful tool for assaying changes in metabolism that affect turgor, such as photosynthetic and osmoregulatory processes. We have developed an updated and automated pressure nephelometer that utilizes visible-infrared (Vis-IR) spectra to accurately quantify GV critical collapse pressure, critical collapse pressure distribution, and cell turgor pressure. Here, using the updated pressure nephelometer and axenic cultures of Microcystis aeruginosa PCC7806, we demonstrate that GV critical collapse pressure is stable during mid-exponential growth phase, introduce pressure-sensitive turbidity as a robust metric for the abundance of gas-vacuolate cyanobacteria, and demonstrate that pressure-sensitive turbidity is a more accurate proxy for abundance and growth than photopigment fluorescence. As cyanobacterium-dominated harmful algal bloom (cyanoHAB) formation is dependent on the constituent cells possessing gas vesicles, characterization of environmental cyanobacteria populations via pressure nephelometry is identified as an underutilized monitoring method. Applications of this instrument focus on physiological and ecological studies of cyanobacteria, for example, cyanoHAB dynamics and the drivers associated with cyanotoxin production in aquatic ecosystems.IMPORTANCE The increased prevalence of bloom-forming cyanobacteria and associated risk of exposure to cyanobacterial toxins through drinking water utilities and recreational waterways are growing public health concerns. Cost-effective, early-detection methodologies specific to cyanobacteria are crucial for mitigating these risks, with a gas vesicle-specific signal offering a number of benefits over photopigment fluorescence, including improved detection limits and discrimination against non-gas-vacuolate phototrophs. Here, we present a multiplexed instrument capable of quantifying the relative abundance of cyanobacteria based on the signal generated from the presence of intracellular gas vesicles specific to bloom-forming cyanobacteria. Additionally, as cell turgor can be measured in vivo via pressure nephelometry, the measurement furnishes information about the internal osmotic pressure of gas-vacuolate cyanobacteria, which relates to the metabolic state of the cell. Together these advances may improve routine waterway monitoring and the mitigation of human health threats due to cyanobacterial blooms.


Mitochondrial DNA damage and repair in RPE associated with aging and age-related macular degeneration.
Mitochondrial DNA (mtDNA) damage may be associated with age-related diseases, such as age-related macular degeneration (AMD). The present study was designed to test whether the frequency of mtDNA damage, heteroplasmic mtDNA mutations, and repair capacity correlate with progression of AMD. Macular and peripheral RPE cells were isolated and cultured from human donor eyes with and without AMD. The stages of AMD were graded according to the Minnesota Grading System. Confluent primary RPE cells were used to test the frequency of endogenous mtDNA damage by quantitative PCR. Mutation detection kits were used to detect heteroplasmic mtDNA mutation. To test the mtDNA repair capacity, cultured RPE cells were allowed to recover for 3 and 6 hours after exposure to H(2)O(2), and repair was assessed by quantitative PCR. The levels of human OGG1 protein, which is associated with mtDNA repair, were analyzed by Western blot. This study showed that mtDNA damage increased with aging and that more lesions occurred in RPE cells from the macular region than the periphery. Furthermore, mtDNA repair capacity decreased with aging, with less mtDNA repair capacity in the macular region compared with the periphery in samples from aged subjects. Most interestingly, the mtDNA damage was positively correlated with the grading level of AMD, whereas repair capacity was negatively correlated. In addition, more mitochondrial heteroplasmic mutations were detected in eyes with AMD. These data show macula-specific increases in mtDNA damage, heteroplasmic mutations, and diminished repair that are associated with aging and AMD severity.


Analysis of the T Cell Response to Zika Virus and Identification of a Novel CD8+ T Cell Epitope in Immunocompetent Mice.
Zika virus (ZIKV) is an emerging arbovirus of the Flaviviridae family. Although ZIKV infection is typically mild and self-limiting in healthy adults, infection has been associated with neurological symptoms such as Guillain-Barré syndrome, and a causal link has been established between fetal microcephaly and ZIKV infection during pregnancy. These risks, and the magnitude of the ongoing ZIKV pandemic, have created an urgent need for the development of animal models to study the immune response to ZIKV infection. Previous animal models have primarily focused on pathogenesis in immunocompromised mice. In this study, we provide a model of ZIKV infection in wild-type immunocompetent C57BL/6 mice, and have provided an analysis of the immune response to infection. We evaluated the activation of several innate immune cell types, and studied the kinetics, phenotype, and functionality of T cell responses to ZIKV infection. Our results demonstrate that ZIKV infection is mild in wild-type immunocompetent C57BL/6 mice, resulting in minimal morbidity. Our data establish that at the peak of the adaptive response, antigen-experienced CD4+ T cells polarize to a Th1 phenotype, and antigen-experienced CD8+ T cells exhibit an activated effector phenotype, producing both effector cytokines and cytolytic molecules. Furthermore, we have identified a novel ZIKV CD8+ T cell epitope in the envelope protein that is recognized by the majority of responding cells. Our model provides an important reference point that will help dissect the impact of polymorphisms in the circulating ZIKV strains on the immune response and ZIKV pathogenesis. In addition, the identification of a ZIKV epitope will allow for the design of tetramers to study epitope-specific T cell responses, and will have important implications for the design and development of ZIKV vaccine strategies.


Advances in plasma skin regeneration.
Plasma skin regeneration (PSR) is a novel method of resurfacing that uses plasma energy to create a thermal effect on the skin. PSR is different from lasers, light sources, and ablative lasers in that it is not chromophore dependent and does not vaporize tissue, but leaves a layer of intact, desiccated epidermis that acts as a natural biologic dressing and promotes wound healing and rapid recovery. Histological studies performed on plasma resurfacing patients have confirmed continued collagen production, reduction of elastosis, and progressive skin rejuvenation beyond 1 year after treatment. PSR has received US Food and Drug Administration 510 (k) clearance for treatment of rhytides of the body, superficial skin lesions, actinic keratoses, viral papillomata, and seborrheic keratoses. PSR also has beneficial effects in the treatment of other conditions including dyschromias, photoaging, skin laxity, and acne scars. The safety profile of PSR is excellent, and there have been no reports of demarcation lines in perioral, periorbital, or jawline areas, as can sometimes be observed following CO2 resurfacing. PSR is effective in improving facial and periorbital rhytides and can be used on nonfacial sites, including the hands, neck, and chest. Numerous treatment protocols with variable energy settings allow for individualized treatments and provide the operator with fine control over the degree of injury and length of subsequent recovery time.


A simple method for measuring thickness of the mucus gel layer adherent to rat, frog and human gastric mucosa: influence of feeding, prostaglandin, N-acetylcysteine and other agents.
1. A technique has been developed for measuring thickness of the gastric surface mucus gel layer. Mucosal sections (1.6 mm) were cut from frog and rat stomach and human antrum, mounted transversely and viewed by an inverse microscope (x 200 magnification) under dark field illumination or phase contrast. The mucus layer was readily distinguishable and its dimensions could be recorded by means of an eyepiece graticule. 2. Mean mucus gel thickness in rat, frog was 73, 76, 55 and 192 micrometer respectively. However, there was variation in the average thickness of the gel layer between individual mucosae from the same species (up to twofold). Mucus thickness between adjacent regions of the same mucosal section also varied markedly (up to tenfold). 3. Topical administration of 16,16-dimethylprostaglandin E2 by oral intubation caused a significant increase in thickness in both rat and frog at doses of 5 microgram/ml and 0.5 microgram/ml respectively. Feeding and exposure of the mucosa to N-acetylcysteine (10-20%, w/v) produced variable effects whereas pepsin (1 mg/ml) caused a marked reduction in thickness of the surface gel layer in both rat and frog. 4. The technique provides a rapid and simple method for determining gastrointestinal mucus thickness in relation to mucosal morphology. It is ideally suited for studying the control of mucus secretion and effect of drugs.


Cloning and functional analysis of the TATA binding protein from Sulfolobus shibatae.
Archaea (formerly archaebacteria) comprise a domain of life that is phylogenetically distinct from both Eucarya and Bacteria. Here we report the cloning of a gene from the Archaeon Sulfolobus shibatae that encodes a protein with strong homology to the TATA binding protein (TBP) of eukaryotes. Sulfolobus shibatae TBP is, however, almost as diverged from other archaeal TBPs that have been cloned as it is from eukaryotic TBPs. DNA binding studies indicate that S.shibatae TBP recognizes TATA-like A-box sequences that are present upstream of most archaeal genes. By quantitatively immunodepleting S.shibatae TBP from an in vitro transcription system, we demonstrate that Sulfolobus RNA polymerase is capable of transcribing the 16S/23S rRNA promoter weakly in the absence of TBP. Most significantly, we show that addition of recombinant S.shibatae TBP to this immunodepleted system leads to transcriptional stimulation and that this stimulation is dependent on the A-box sequence of the promoter. Taken together, these findings reveal fundamental similarities between the transcription machineries of Archaea and eukaryotes.


Mycobacterial DNA extraction for whole-genome sequencing from early positive liquid (MGIT) cultures.
We developed a low-cost and reliable method of DNA extraction from as little as 1 ml of early positive mycobacterial growth indicator tube (MGIT) cultures that is suitable for whole-genome sequencing to identify mycobacterial species and predict antibiotic resistance in clinical samples. The DNA extraction method is based on ethanol precipitation supplemented by pretreatment steps with a MolYsis kit or saline wash for the removal of human DNA and a final DNA cleanup step with solid-phase reversible immobilization beads. The protocol yielded ≥0.2 ng/μl of DNA for 90% (MolYsis kit) and 83% (saline wash) of positive MGIT cultures. A total of 144 (94%) of the 154 samples sequenced on the MiSeq platform (Illumina) achieved the target of 1 million reads, with <5% of reads derived from human or nasopharyngeal flora for 88% and 91% of samples, respectively. A total of 59 (98%) of 60 samples that were identified by the national mycobacterial reference laboratory (NMRL) as Mycobacterium tuberculosis were successfully mapped to the H37Rv reference, with >90% coverage achieved. The DNA extraction protocol, therefore, will facilitate fast and accurate identification of mycobacterial species and resistance using a range of bioinformatics tools.


Emotional support predicts more sickness absence and poorer self assessed work ability: a two-year prospective cohort study.
While back pain and stressful work environment are shown to be important causes of sickness absence the effect of psychosocial resources on sickness absence, and on self assessed work ability, is less commonly investigated. The aim of this study was to assess these associations in a two-year follow-up study. 341 working people aged 45 to 64, randomly drawn from the population, responded to a questionnaire at baseline and at a two-year follow-up. Poisson regression was used to analyse the association of psychosocial factors (psychosocial instruments on work environment, emotional support and psychological resources) and previous back pain (low back and/or neck) at baseline with sickness absence (spells and days) at follow-up, controlling for effects of age, sex, BMI, smoking, alcohol, occupation, disease and previous sickness absence. Logistic regression was used to study the associations of psychosocial factors and previous back pain at baseline with self assessed prognosis of poor work ability six months from follow-up. Finally, a multivariate analysis tested the independent effects of previous back pain and 3 psychosocial factors derived in a factor analysis: 1. work environment; 2. emotional support; 3. psychological resources, on work ability and absence days and spells. 80% of the sickness absence spells within the last 12 months before follow-up were short-term (≤ 14 days). In the final model, high emotional support predicted more sickness absence spells (RR 1.36; 1.11-1.67) and days (RR 1.68, 1.22-2.31). Previous back pain (OR 2.56; 1.13-5.81), high emotional support (OR 1.58; 1.02-2.46), and low psychological resources (OR 0.62; 0.44-0.89) were related to poorer self assessed prognosis of work ability at follow up. In a general middle aged working population high emotional support was related to more sickness absence and also poorer self assessed prognosis of work ability. Our findings suggest that both sickness absence and self assessed work ability are dependent of life outside work and can be affected by a person's close community.


Developing novel prognostic biomarkers for multivariate fracture risk prediction algorithms.
Multivariate prediction algorithms such as FRAX® and QFractureScores provide an opportunity for new prognostic biomarkers to be developed and incorporated, potentially leading to better fracture prediction. As more research is conducted into these novel biomarkers, a number of factors need to be considered for their successful development for inclusion in these algorithms. In this review, we describe two well-known multivariate prediction algorithms for osteoporosis fracture risk applicable to the UK population, FRAX and QFractureScores, and comment on the current prognostic tools available for fracture risk; dual X-ray assessment, quantitative ultrasonography, and genomic/biochemical markers. We also highlight the factors that need to be considered in the development of new biomarkers. These factors include the requirement for prospective data, collected in new cohort studies or using archived samples; the need for adequate stability data to be provided; and the need for appropriate storage methods to be used when retrospective data are required. Area under the receiver operating characteristic curve measures have been found to have limited utility in assessing the impact of the addition of new risk factors on the predictive performance of multivariate algorithms. New performance evaluation measures, such as net reclassification index and integrated discrimination improvement, are increasingly important in the evaluation of the impact of the addition of new markers to multivariate algorithms, and these are also discussed.


Shaping successful and unsuccessful CD8 T cell responses following infection.
CD8 T cells play a vital role in the immunological protection against intracellular pathogens. Ideally, robust effector responses are induced, which eradicate the pathogen, and durable memory CD8 T cells are also established, which help confer protection against subsequent reinfection. The quality and magnitude of these responses is dictated by multiple factors, including their initial interactions with professional antigen-presenting cells, as well as the cytokine milieu and availability of CD4 T cell help. These factors set the transcriptional landscape of the responding T cells, which in turn influences their phenotypic and functional attributes as well as ultimate fate. Under certain conditions, such as during chronic infections, the development of these usually successful responses becomes subverted. Here we discuss advances in our understanding of the cellular and molecular determinants of T cell quality, and the formation of effector, memory, and exhausted CD8 T cells, during acute and chronic infections.


Tracking and treating malnutrition: a retrospective observational study of the nutritional status of vulnerable people accessing a meals-on-wheels (MOW) service.
The aims of the study were to describe the characteristics of meals-on-wheels (MOW) recipients, including prevalence of malnutrition amongst those who have received input from the Nutrition and Wellbeing Service (NWS) and to explore whether the NWS had an impact on the nutritional status (malnutrition risk) of recipients over time. Support services, for example, MOW, play an important role in the prevention and treatment of malnutrition in the community. In the UK, MOW services are under threat. However, little is known about how they support the health and well-being of older people. This study reports on the characteristics of MOW recipients and investigates change in nutritional status over time. A retrospective study of MOW recipients of nutritional concern who were offered a check through the NWS was conducted. Demographic, social and health information were gathered at the initial visit. Nutritional status (risk of malnutrition) was obtained using the validated Malnutrition Universal Screening Tool (MUST), at the initial and subsequent visits. Changes over time were investigated for recipients receiving at least two follow-up visits. An initial visit was made to 399 MOW recipients, and 148 recipients had two or more follow-up visits. At initial screening, 177 (44%) of recipients were at medium or high risk of malnutrition. Frailty was significantly related to malnutrition risk (P = 0.049). At follow-up, there was a reduction in malnutrition risk. The MOW service was associated with a reduction in malnutrition risk. By offering well-being visits within a MOW service, malnutrition can be identified early. Future studies into how MOW services might delay or prevent the need for support from acute health services and social care are warranted.


The small heat shock protein HSP27 is not an independent prognostic marker in axillary lymph node-negative breast cancer patients.
Heat shock protein 27 (hsp27) belongs to the family of heat shock proteins and is thought to be involved in thermotolerance, cell proliferation, drug resistance, and chaperone processes. The aim of this study was to investigate whether hsp27 levels are correlated with clinical outcome in axillary lymph node-negative breast cancer patients. We describe a Western blot study measuring hsp27 levels in 425 patients and an immunohistochemistry (IHC) study analyzing 788 patients. Results obtained by both methods were concordant. Univariate survival analysis was performed considering hsp27 either as an optimally dichotomized variable or as a continuous variable. Additional data include age at biopsy, tumor size, estrogen receptor (ER) and progesterone receptor status, tumor ploidy and percentage of cells in S phase, and adjuvant therapy. hsp27 levels correlated positively with ER status (P = 0.0001 in Western blot and IHC study), progesterone receptor status (P = 0.0001 in Western blot and IHC study), and aneuploidy (Western blot study, P = 0.0012; IHC study, P = 0.0004) but not with tumor size (Western blot study, P = 0.69; IHC, P = 0.53) or S phase (Western blot study, P = 0.19; IHC study, P = 0.38). Overall, there was no relationship between hsp27 expression and disease-free survival (Western blot study, P = 0.70/0.54; IHC, P = 0.47/0.30) or overall survival (Western blot study, P = 0.16/0.15; IHC, P = 0.46/0.78). Exploratory subset analyses defined by ER status and use of adjuvant treatment indicated that in ER+/untreated patients, high hsp27 levels correlated modestly with shorter disease-free survival (Western blot, P = 0.04/0.04; IHC, P = 0.11/0. 03). hsp27 is not a useful prognostic marker for the clinic in axillary lymph node-negative patients. However, the finding of modest prognostic value of hsp27 in the subgroup of ER+/untreated patients raises new questions about the biological function of hsp27 in breast cancer.


Small retinal haemorrhages accompanied by macular soft drusen: prevalence, and funduscopic and angiographic characteristics.
To investigate the prevalence and clinical significance of small retinal haemorrhages accompanied by macular soft drusen in exudative age-related macular degeneration (AMD). This observational case series included patients who had first been diagnosed with exudative AMD. Small retinal haemorrhages were defined as preretinal or intraretinal haemorrhages, no larger than half the disc diameter in size and located within 3000 μm of the fovea centre. If there was more than one haemorrhage, the entire affected area was less than two-thirds of the disc diameter. Macular soft drusen was defined as the presence of soft drusen (≥125 μm in diameter) within the macular area. The presence of retinal angiomatous proliferation (RAP) was estimated based on the results of indocyanine green angiography (ICGA). The prevalence of reticular pseudodrusen was also estimated. Among the 1921 eyes from 1604 patients who were newly diagnosed with exudative AMD during the 40 months prior to the study, 101 eyes (5.3%) from 79 patients presented with the fundus characteristics described above. ICGA images were available for 69 eyes. Among these eyes, 28 eyes (43.1%) and 25 eyes (38.5%) were found to have type 1 and 2 RAP, respectively. A chorioretinal anastomosis (type 3 RAP) was identified in 12 (18.5%) eyes. Reticular pseudodrusen were noted in 78 eyes (77.2%). The presence of small retinal haemorrhages accompanied by macular soft drusen was highly predictive of RAP. The high prevalence of both soft drusen and reticular pseudodrusen in these eyes may suggest a profound decrease in choroidal perfusion in these eyes.


Effect of short-term systemic glucocorticoid treatment on human nasal mediator release after antigen challenge.
The effect of systemic glucocorticoid treatment on early- and late-phase nasal allergic reactions after allergen challenge was determined in a double-blind, cross-over study in 13 allergic individuals. The subjects were pretreated for 2 d before challenge with 60 mg prednisone per day or a matching placebo. A previously described model using repeated nasal lavages for measuring mediator release in vivo was utilized. Symptom scores obtained repeatedly before, during, and after the challenge and the number and timing of sneezes were recorded. The mediators measured were histamine. N-alpha-p-tosyl-L-arginine methyl ester (TAME)-esterase activity, kinins, PGD2, and LTC4/D4. Albumin was also measured as a marker of plasma transudation. Blood samples were taken for determination of total number of white blood cells, differential count, and total blood histamine content. No effect of steroid therapy was found on the appearance of symptoms or any of the mediators, except a reduction in kinins, in the early phase of the allergic reaction. However, in the late phase, the prednisone reduced the number of sneezes (P less than 0.01), as well as the level of histamine (P less than 0.05), TAME-esterase activity (P less than 0.05), kinins (P less than 0.05), and albumin (P less than 0.05). Only low levels of leukotrienes were found in the late phase, but the quantities of these mediators seemed to be decreased by the glucocorticoid treatment (P = 0.06). PGD2 did not increase during the LPR and thus was not affected by glucocorticosteroids. The immediate response to a second challenge 11 h after the first was also evaluated. Whereas the appearance of mediators was enhanced over the initial response to the same challenge dose in placebo-treated subjects, this enhancement was abrogated after prednisone treatment. As this dose of drug is known to be clinically effective in treating hay fever, the present study confirms the earlier findings of others that short-term systemic glucocorticoid treatment inhibits the late phase but not the immediate phase of antigen challenge. Furthermore, secondary enhancement of immediate responses is inhibited. This study shows that glucocorticoids inhibit the generation or release of inflammatory mediators during the late reaction and the physiologic response.


Amide bond formation: beyond the myth of coupling reagents.
Amide bond formation is a fundamentally important reaction in organic synthesis, and is typically mediated by one of a myriad of so-called coupling reagents. This critical review is focussed on the most recently developed coupling reagents with particular attention paid to the pros and cons of the plethora of "acronym" based reagents. It aims to demystify the process allowing the chemist to make a sensible and educated choice when carrying out an amide coupling reaction (179 references).


Peptides derived from the auxin binding protein elevate Ca2+ and pH in stomatal guard cells of Vicia faba: a confocal fluorescence ratio imaging study.
Dual-excitation confocal laser scanning microscopy (CLSM) was used to image the pH-indicator, BCECF, iontophoretically microinjected into stomatal guard cells of Vicia faba during challenge with peptides derived from hydrophilic domains of the maize auxin-binding protein. Only the peptide corresponding to the C-terminal end (Pz151-163) caused significant changes in cytosolic pH, stimulating rapid alkalinisation of 0.4 +/- 0.1 pH units. Cytosolic pH was clamped using the permeant weak acid, butyrate, and this treatment buffered the peptide evoked alkalinisation. In concert with the electrical events monitored at the plasma membrane using whole-cell voltage clamp, this provides strong evidence for a role of [H+] as a signal intermediate in the guard cell transduction network. In preliminary experiments using single-wavelength imaging of the calcium-indicator, Fluo-3, Pz151-163 also stimulated rapid, reversible increases in cytosolic calcium, whilst two other peptides tested had no effect.


Toxicological profile of ultrapure 2,2',3,4,4',5,5'-heptachlorbiphenyl (PCB 180) in adult rats.
PCB 180 is a persistent non-dioxin-like polychlorinated biphenyl (NDL-PCB) abundantly present in food and the environment. Risk characterization of NDL-PCBs is confounded by the presence of highly potent dioxin-like impurities. We used ultrapure PCB 180 to characterize its toxicity profile in a 28-day repeat dose toxicity study in young adult rats extended to cover endocrine and behavioral effects. Using a loading dose/maintenance dose regimen, groups of 5 males and 5 females were given total doses of 0, 3, 10, 30, 100, 300, 1000 or 1700 mg PCB 180/kg body weight by gavage. Dose-responses were analyzed using benchmark dose modeling based on dose and adipose tissue PCB concentrations. Body weight gain was retarded at 1700 mg/kg during loading dosing, but recovered thereafter. The most sensitive endpoint of toxicity that was used for risk characterization was altered open field behavior in females; i.e. increased activity and distance moved in the inner zone of an open field suggesting altered emotional responses to unfamiliar environment and impaired behavioral inhibition. Other dose-dependent changes included decreased serum thyroid hormones with associated histopathological changes, altered tissue retinoid levels, decreased hematocrit and hemoglobin, decreased follicle stimulating hormone and luteinizing hormone levels in males and increased expression of DNA damage markers in liver of females. Dose-dependent hypertrophy of zona fasciculata cells was observed in adrenals suggesting activation of cortex. There were gender differences in sensitivity and toxicity profiles were partly different in males and females. PCB 180 adipose tissue concentrations were clearly above the general human population levels, but close to the levels in highly exposed populations. The results demonstrate a distinct toxicological profile of PCB 180 with lack of dioxin-like properties required for assignment of WHO toxic equivalency factor. However, PCB 180 shares several toxicological targets with dioxin-like compounds emphasizing the potential for interactions.


Age-related response to interferon alfa treatment in women vs men with chronic hepatitis C virus infection.
Interferon alfa is used widely for patients with chronic hepatitis C virus (HCV) infection. Little is known, however, of the relationship between patients' sex and the effectiveness of interferon alfa treatment in these patients. We treated 311 patients (199 men and 112 women) with human lymphoblastoid interferon (6 million units subcutaneously every day for 2 weeks and 3 times a week for 22 weeks) and observed them for an additional 6 months. Serum HCV RNA levels and genotype were tested by polymerase chain reaction before treatment. A liver biopsy was also done. For the purposes of this study, a complete response was defined as the elimination of HCV RNA for at least 6 months after the termination of treatment. The rate of complete response was 27.1% for men and 24.1% for women. With multiple logistic regression analysis, the HCV RNA level (P < .001), genotype (P < .001), patients' sex (P < .05), and the interaction between sex and age were associated with a complete response to interferon alfa. The rate of complete response was 33.3% in men aged 39 years and younger, 25.0% in men aged 40 years and older. 75.0% in women aged 39 years and younger, and 15.6% in women aged 40 years and older. The odds ratio by group was 1.00, 0.72, 4.38, and 0.21, respectively. Our finding that women aged 39 years and younger are responsive to interferon alfa treatment suggests that hormonal activity, in particular the level of estrogen, may be associated with the sustained elimination of HCV.


Psychosocial effects of workplace exercise - A systematic review.
Objectives Workplace exercise interventions are known to reduce musculoskeletal complaints and are widely offered to promote employee`s health. The psychosocial effects of such interventions are not clear. This review evaluates the effects of workplace exercise interventions on the psychosocial work environment and mental health. Methods A systematic review was performed by screening four scientific databases (Pubmed, Cochrane Database, PsycINFO and Web of Science) for published randomized controlled trials in English from 1998-2018. The review followed the PRISMA statement guidelines using the Cochrane tool for assessing risk of bias and the GRADE approach for assessment of quality. Two researchers independently performed database search, study screening, and evaluation. Results Twenty two randomized controlled trials met the inclusion criteria. Substantial heterogenicity in the studies included was found in study populations, types of intervention, and outcome measures performed. Quality assessment revealed a generally low level of quality in the studies included. This was partly the result of the limited possibilities for blinding and the self-reported outcome measures. Conclusion Workplace exercise interventions seem to have limited effects on mental health and the psychosocial work environment. Future studies should incorporate blinding and independent outcome measures.


Amiodarone induced "Blue man syndrome"; an unusual presentation.
Amiodarone is a common antiarrhythmic medication used in daily practice with excellent efficiency. While it has many benefits, there are several common adverse reactions, such as abnormal thyroid function and cornea verticillata, associated with its use. A much less common adverse reaction is the so-called "Blue man syndrome," occurring in 1-3% of patients. Blue man syndrome is thought to stem from the deposition of lysosomal membrane-bound dense bodies, similar to lipofuscin, in the dermis of patients on chronic amiodarone therapy. We present the case of a 77-year-old male who presented to us post-cardiac arrest secondary to hyperkalemia of 7.0. Although it was noted in the patient's chart that he had an "allergy" to amiodarone (it was noted in his chart it only turned his leg black), it was used in the emergency department because of the failure of other antiarrhythmics to treat his arrhythmia and all other medical options were exhausted. It was decided that the benefits of using amiodarone far outweighed the risks of it. Ultimately it was found that the patient did not have a true allergy to amiodarone, but suffered from a known benign adverse effect of the drug resulting in a greyish-blue tinge to his bilateral lower extremities. His-presentation differs from the normal presentation of blue man syndrome which appears more commonly on sun-exposed areas of the body such as the face and arms. While this is a benign adverse effect of amiodarone, it can be very distressing to patients and must be addressed.


Orthodontic treatment for prominent upper front teeth (Class II malocclusion) in children and adolescents.
Prominent upper front teeth are a common problem affecting about a quarter of 12-year-old children in the UK. The condition develops when permanent teeth erupt. These teeth are more likely to be injured and their appearance can cause significant distress. Children are often referred to an orthodontist for treatment with dental braces to reduce the prominence of their teeth. If a child is referred at a young age, the orthodontist is faced with the dilemma of whether to treat the patient early or to wait and provide treatment in adolescence. To assess the effects of orthodontic treatment for prominent upper front teeth initiated when children are seven to 11 years old ('early treatment' in two phases) compared to in adolescence at around 12 to 16 years old ('late treatment' in one phase); to assess the effects of late treatment compared to no treatment; and to assess the effects of different types of orthodontic braces. Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 27 September 2017), the Cochrane Central Register of Controlled Trials (CENTRAL) (the Cochrane Library, 2017, Issue 8), MEDLINE Ovid (1946 to 27 September 2017), and Embase Ovid (1980 to 27 September 2017). The US National Institutes of Health Ongoing Trials Registry (ClinicalTrials.gov) and the World Health Organization International Clinical Trials Registry Platform were searched for ongoing trials. No restrictions were placed on the language or date of publication when searching the electronic databases. Randomised controlled trials of orthodontic treatments to correct prominent upper front teeth (Class II malocclusion) in children and adolescents. We included trials that compared early treatment in children (two-phase) with any type of orthodontic braces (removable, fixed, functional) or head-braces versus late treatment in adolescents (one-phase) with any type of orthodontic braces or head-braces, and trials that compared any type of orthodontic braces or head-braces versus no treatment or another type of orthodontic brace or appliance (where treatment started at a similar age in the intervention groups).We excluded trials involving participants with a cleft lip or palate, or other craniofacial deformity/syndrome, and trials that recruited patients who had previously received surgical treatment for their Class II malocclusion. Review authors screened the search results, extracted data and assessed risk of bias independently. We used odds ratios (ORs) and 95% confidence intervals (CIs) for dichotomous outcomes, and mean differences (MDs) and 95% CIs for continuous outcomes. We used the fixed-effect model for meta-analyses including two or three studies and the random-effects model for more than three studies. We included 27 RCTs based on data from 1251 participants.Three trials compared early treatment with a functional appliance versus late treatment for overjet, ANB and incisal trauma. After phase one of early treatment (i.e. before the other group had received any intervention), there was a reduction in overjet and ANB reduction favouring treatment with a functional appliance; however, when both groups had completed treatment, there was no difference between groups in final overjet (MD 0.21, 95% CI -0.10 to 0.51, P = 0.18; 343 participants) (low-quality evidence) or ANB (MD -0.02, 95% CI -0.47 to 0.43; 347 participants) (moderate-quality evidence). Early treatment with functional appliances reduced the incidence of incisal trauma compared to late treatment (OR 0.56, 95% CI 0.33 to 0.95; 332 participants) (moderate-quality evidence). The difference in the incidence of incisal trauma was clinically important with 30% (51/171) of participants reporting new trauma in the late treatment group compared to only 19% (31/161) of participants who had received early treatment.Two trials compared early treatment using headgear versus late treatment. After phase one of early treatment, headgear had reduced overjet and ANB; however, when both groups had completed treatment, there was no evidence of a difference between groups in overjet (MD -0.22, 95% CI -0.56 to 0.12; 238 participants) (low-quality evidence) or ANB (MD -0.27, 95% CI -0.80 to 0.26; 231 participants) (low-quality evidence). Early (two-phase) treatment with headgear reduced the incidence of incisal trauma (OR 0.45, 95% CI 0.25 to 0.80; 237 participants) (low-quality evidence), with almost half the incidence of new incisal trauma (24/117) compared to the late treatment group (44/120).Seven trials compared late treatment with functional appliances versus no treatment. There was a reduction in final overjet with both fixed functional appliances (MD -5.46 mm, 95% CI -6.63 to -4.28; 2 trials, 61 participants) and removable functional appliances (MD -4.62, 95% CI -5.33 to -3.92; 3 trials, 122 participants) (low-quality evidence). There was no evidence of a difference in final ANB between fixed functional appliances and no treatment (MD -0.53°, 95% CI -1.27 to -0.22; 3 trials, 89 participants) (low-quality evidence), but removable functional appliances seemed to reduce ANB compared to no treatment (MD -2.37°, 95% CI -3.01 to -1.74; 2 trials, 99 participants) (low-quality evidence).Six trials compared orthodontic treatment for adolescents with Twin Block versus other appliances and found no difference in overjet (0.08 mm, 95% CI -0.60 to 0.76; 4 trials, 259 participants) (low-quality evidence). The reduction in ANB favoured treatment with a Twin Block (-0.56°, 95% CI -0.96 to -0.16; 6 trials, 320 participants) (low-quality evidence).Three trials compared orthodontic treatment for adolescents with removable functional appliances versus fixed functional appliances and found a reduction in overjet in favour of fixed appliances (0.74, 95% CI 0.15 to 1.33; two trials, 154 participants) (low-quality evidence), and a reduction in ANB in favour of removable appliances (-1.04°, 95% CI -1.60 to -0.49; 3 trials, 185 participants) (low-quality evidence). Evidence of low to moderate quality suggests that providing early orthodontic treatment for children with prominent upper front teeth is more effective for reducing the incidence of incisal trauma than providing one course of orthodontic treatment in adolescence. There appear to be no other advantages of providing early treatment when compared to late treatment. Low-quality evidence suggests that, compared to no treatment, late treatment in adolescence with functional appliances, is effective for reducing the prominence of upper front teeth.


New developments in treatment after lung transplantation.
Lung transplantation has evolved as an accepted therapy in selected adults and children with end-stage lung disease. Outcomes following lung transplantation have improved in the recent era with a 5-year survival of > 70% and an overall good functional status of surviving recipients. Many of the advances have been achieved by the use of modern immunosuppressive agents. To date, multiple strategies exist that may be employed when utilizing immunosuppression. These agents can be used in a variety of roles that may include induction, maintenance or rescue therapy, many of which are illustrated in this review including the current evidence to support their use. Infections in lung transplant recipients remain a significant cause of morbidity and mortality. Special considerations are required with the substantial burden of chronic infection in candidates with CF lung disease before transplantation, which are discussed. Furthermore, recent progress and advances in prevention and treatment of post-transplantation infectious complications are detailed. Chronic lung allograft dysfunction remains to be the burden of lung transplantation in the long-term. Unfortunately, there is no well-established therapy to address it. However, therapy attempts include change/augmentation of immunosupression, use of neomacrolides and extracorporeal photopheresis, all of which are reviewed in detail.


Adenovirus respiratory tract infections in Peru.
Currently, there is a paucity of data regarding human adenovirus (HAdv) circulation in Andean regions of South America. To address this shortcoming, we report the clinical, phylogenetic, and epidemiologic characteristics of HAdv respiratory tract infection from a large sentinel surveillance study conducted among adults and children in Peru. Oropharyngeal swabs were collected from participants visiting any of 38 participating health centers, and viral pathogens were identified by immunofluorescence assay in cell culture. In addition, molecular characterization was performed on 226 randomly selected HAdv samples. Between 2000 and 2010, a total of 26,375 participants with influenza-like illness (ILI) or severe acute respiratory infection (SARI) were enrolled in the study. HAdv infection was identified in 2.5% of cases and represented 6.2% of all viral pathogens. Co-infection with a heterologous virus was found in 15.5% of HAdv cases. HAdv infection was largely confined to children under the age of 15, representing 88.6% of HAdv cases identified. No clinical characteristics were found to significantly distinguish HAdv infection from other respiratory viruses. Geographically, HAdv infections were more common in sites from the arid coastal regions than in the jungle or highland regions. Co-circulation of subgroups B and C was observed each year between 2006 and 2010, but no clear seasonal patterns of transmission were detected. HAdv accounted for a significant fraction of those presenting with ILI and SARI in Peru and tended to affect the younger population disproportionately. Longitudinal studies will help better characterize the clinical course of patients with HAdv in Peru, as well as determine the role of co-infections in the evolution of illness.


Parental rheumatoid arthritis and long-term child morbidity: a nationwide cohort study.
To estimate the influence of parental rheumatoid arthritis (RA) on child morbidity. Nationwide cohort study. Individual linkage to nationwide Danish registries. All singletons born in Denmark during 1977-2008 (n=1 917 723) were followed for an average of 16 years. Adjusted HRs for child morbidity; that is, 11 main diagnostic groups and specific autoimmune diseases within the International Classification of Diseases 8th and 10th versions. Compared with unexposed children, children exposed to maternal RA ('clinical' and 'preclinical') (n=13 566) had up to 26% higher morbidity in 8 of 11 main diagnostic groups. Similar tendencies were found in children exposed to paternal RA ('clinical' and 'preclinical') (n=6330), with statistically significantly higher morbidity in 6 of 11 diagnostic groups. HRs were highest for autoimmune diseases with up to three times increased risk of juvenile idiopathic arthritis (HR, 95% CI 3.30, 2.71 to 4.03 and 2.97, 2.20 to 4.01) and increased risk of up to 40% of diabetes mellitus type 1 (HR, 95% CI 1.37, 1.12 to 1.66 and 1.44, 1.09 to 1.90) and up to 30% increased HR of asthma (HR, 95% CI 1.28, 1.20 to 1.36 and 1.15, 1.04 to 1.26). Conclusions were roughly similar for children exposed to maternal clinical RA and for children only followed up to 16 years of age. Children of parents with RA had consistent excess morbidity. If the associations reflect biological mechanisms, genetic factors seem to play an important role. These findings call for attention given to children of parents with RA.


Three-dimensional kinematics and limb kinetic energy of running cockroaches.
We tested the hypothesis that fast-running hexapeds must generate high levels of kinetic energy to cycle their limbs rapidly compared with bipeds and quadrupeds. We used high-speed video analysis to determine the three-dimensional movements of the limbs and bodies of cockroaches (Blaberus discoidalis) running on a motorized treadmill at 21 cm s-1 using an alternating tripod gait. We combined these kinematic data with morphological data to calculate the mechanical energy produced to move the limbs relative to the overall center of mass and the mechanical energy generated to rotate the body (head + thorax + abdomen) about the overall center of mass. The kinetic energy involved in moving the limbs was 8 microJ stride-1 (a power output of 21 mW kg-1, which was only approximately 13% of the external mechanical energy generated to lift and accelerate the overall center of mass at this speed. Pitch, yaw and roll rotational movements of the body were modest (less than +/- 7 degrees), and the mechanical energy required for these rotations was surprisingly small (1.7 microJ stride-1 for pitch, 0.5 microJ stride-1 for yaw and 0.4 microJ stride-1 for roll) as was the power (4.2, 1.2 and 1.1 mW kg-1, respectively). Compared at the same absolute forward speed, the mass-specific kinetic energy generated by the trotting hexaped to swing its limbs was approximately half of that predicted from data on much larger two- and four-legged animals. Compared at an equivalent speed (mid-trotting speed), limb kinetic energy was a smaller fraction of total mechanical energy for cockroaches than for large bipedal runners and hoppers and for quadrupedal trotters. Cockroaches operate at relatively high stride frequencies, but distribute ground reaction forces over a greater number of relatively small legs. The relatively small leg mass and inertia of hexapeds may allow relatively high leg cycling frequencies without exceptionally high internal mechanical energy generation.


The coronary circulation in hypertensive heart disease.
This study analyzes and compares systemic and coronary hemodynamics in patients with essential hypertension in relation to hypertrophic heart disease of nonhypertensive origin. Left ventricular function (as assessed from the cardiac index, stroke volume index, ejection fraction, mean velocity of circumferential fiber shortening, mean normalized systolic ejection rate, and isovolumic indexes) may be normal in patients with hypertensive hypertrophy, even with a large increase in muscle mass and in the presence of concomitant coronary artery disease. Left ventricular function is impaired when regional contraction abnormalities or ventricular dilatation, or both, occur and is inversely related to both cardiac size and systolic wall stress. Coronary blood flow (+ 18%), coronary resistance (+ 38%), and myocardial oxygen consumption (MVO2) (+ 21%) are increased in essential hypertension. Coronary reserve is reduced even in hypertensive hypertrophy without evidence of coronary artery disease. MVO2 per mass unit was directly correlated with systolic wall stress per cross-sectional area of the left ventricular wall. Coronary reserve may remain normal in both moderate and excessive hypertrophy, provided systolic wall stress and hence the myocardial oxygen consumption are not increased. It is concluded that the appropriateness of left ventricular hypertrophy, as a result of mass-to-volume ratio and stress, is a major determinant of left ventricular performance, of coronary blood flow, and of myocardial oxygen consumption.


Tuberculosis in patients with human immunodeficiency virus infection.
Tuberculosis (TB) is the major opportunistic infection of human immunodeficiency virus (HIV)-infected persons worldwide. Human immunodeficiency virus infection is the most potent known risk factor for reactivation of latent Mycobacterium tuberculosis infection, and TB disease appears to increase the rate of HIV progression. Pulmonary disease is seen in most patients, including a large proportion of those with extrapulmonary disease. Failure to suspect TB and to order the appropriate diagnostic tests is the most common reason for diagnostic delays. With advancing HIV infection, tuberculin skin test reactivity decreases along with reactivity to nonspecific antigens such as mumps, tetanus toxoid, and Candida; anergy testing need not be a routine component of tuberculosis screening of HIV-infected persons. The diagnosis depends on identifying the organism on smears or cultures; direct amplification tests may facilitate rapid identification of M. tuberculosis, but the relatively low sensitivity in smear-negative specimens limits their use. Also, these tests must be used in conjunction with the clinical assessment, and they must always be performed in conjunction with microscopy and standard culture. Shorter courses of combination preventive therapy of patients with latent tuberculous infection are effective, but the potential advantages of improved adherence and reduced costs of shorter courses should be balanced with an increased risk secondary to ongoing TB exposure in areas with a high TB prevalence. Six months of treatment for active tuberculosis is recommended, unless the response of a particular patient is slow or otherwise suboptimal. The use of highly active antiretroviral therapy (HAART) made a remarkable impact on the course or HIV disease, but raises several issues with respect to HIV-related TB. Drug interactions necessitate either a non-rifamycin-based regimen or a rifabutin-based regimen in patients on HAART treated for TB.


The Pathway for Oxygen: Tutorial Modelling on Oxygen Transport from Air to Mitochondrion: The Pathway for Oxygen.
The 'Pathway for Oxygen' is captured in a set of models describing quantitative relationships between fluxes and driving forces for the flux of oxygen from the external air source to the mitochondrial sink at cytochrome oxidase. The intervening processes involve convection, membrane permeation, diffusion of free and heme-bound O2 and enzymatic reactions. While this system's basic elements are simple: ventilation, alveolar gas exchange with blood, circulation of the blood, perfusion of an organ, uptake by tissue, and consumption by chemical reaction, integration of these pieces quickly becomes complex. This complexity led us to construct a tutorial on the ideas and principles; these first PathwayO2 models are simple but quantitative and cover: (1) a 'one-alveolus lung' with airway resistance, lung volume compliance, (2) bidirectional transport of solute gasses like O2 and CO2, (3) gas exchange between alveolar air and lung capillary blood, (4) gas solubility in blood, and circulation of blood through the capillary syncytium and back to the lung, and (5) blood-tissue gas exchange in capillaries. These open-source models are at Physiome.org and provide background for the many respiratory models there.


Fault diagnosis of a benchmark fermentation process: a comparative study of feature extraction and classification techniques.
This paper investigates fault diagnosis in batch processes and presents a comparative study of feature extraction and classification techniques applied to a specific biotechnological case study: the fermentation process model by Birol et al. (Comput Chem Eng 26:1553-1565, 2002), which is a benchmark for advanced batch processes monitoring, diagnosis and control. Fault diagnosis is achieved using four approaches on four different process scenarios based on the different levels of noise so as to evaluate their effects on the performance. Each approach combines a feature extraction method, either multi-way principal component analysis (MPCA) or multi-way independent component analysis (MICA), with a classification method, either artificial neural network (ANN) or support vector machines (SVM). The performance obtained by the different approaches is assessed and discussed for a set of simulated faults under different scenarios. One of the faults (a loss in mixing power) could not be detected due to the minimal effect of mixing on the simulated data. The remaining faults could be easily diagnosed and the subsequent discussion provides practical insight into the selection and use of the available techniques to specific applications. Irrespective of the classification algorithm, MPCA renders better results than MICA, hence the diagnosis performance proves to be more sensitive to the selection of the feature extraction technique.


T cell responses to 53-kDa outer membrane protein of Porphyromonas gingivalis in humans with early-onset periodontitis.
Patients with early-onset periodontitis (EOP) are susceptible to infection with periodontopathic bacteria, such as Porphyromonas gingivalis. Ag53, 53-kDa outer membrane protein of P. gingivalis, evokes strong humoral immune responses in EOP patients. In a first step to clarify how host immune cells recognize Ag53, we established Ag53-specific short-term T cell lines from 22 subjects including 6 EOP patients and 16 healthy donors, using overlapping peptides based on Ag53 amino acid sequences. All T cell lines from active EOP patients recognized a common region (p141-181, especially p141-161) on Ag53, while those from healthy donors showed heterogeneous specificity. p141-181 was not recognized by T cell lines established from EOP patients following therapy. A monoclonal antibody to HLA-DRB 1 inhibited Ag53-induced proliferation of most of the T cell lines. Our observations suggest that, although antigen-presenting molecules are common in EOP patients and in healthy individuals, p141-161 includes a major T cell epitope(s) on Ag53 for active EOP patients but not for healthy individuals or inactive EOP patients.


Case series on acute HCV in HIV-negative men in regular clinical practice: a call for action.
The evidence that HIV treatment as prevention (TasP) and HIV pre-exposure prophylaxis (PrEP) reduces the risk of HIV transmission is overwhelming. But as PrEP and TasP can lead to increased sexual mixing between HIV positive and negative men who have sex with men (MSM), sexually transmitted infections such as acute hepatitis C (HCV), which were thought to be limited to HIV-infected MSM, could become more frequent in HIV uninfected MSM as well. The objective of this study was to describe a series of cases of sexually transmitted HCV infections in HIV-uninfected MSM in the Netherlands and Belgium. Through the Dutch Acute HCV in HIV Study (a Dutch-Belgian prospective multicentre study on the treatment of acute HCV infection, NCT02600325) and the Be-PrEP-ared study (a PrEP project in Antwerp, EudraCT2015-000054-37) several acute HCV infections were detected in HIV-negative men. A newly acquired HCV infection was diagnosed in ten HIV-negative MSM. HCV was diagnosed at a sexually transmitted infection (STI) clinic (n = 2), by their general practitioner (n = 2), by their HIV physician (n = 1) or at a PrEP clinic (n = 5). Ten patients reported unprotected anal intercourse and four had a concomitant STI at the time of HCV diagnosis. Six patients reported using drugs during sex. Our observation calls for a larger nationwide epidemiological study on the prevalence, incidence and risk factors of HCV infection in HIV-uninfected MSM. In the changing landscape of TasP and PrEP, reliable and up-to-date epidemiological data on HCV among HIV-uninfected MSM are needed and will help in developing evidence-based testing policies.


"I Think That I Have a Good Understanding of How to Protect Myself": A Qualitative Study About HIV Risk Perceptions Among Men and Gender Diverse Individuals Who Have Sex With Men.
This qualitative study aimed to determine how men and gender diverse individuals who have sex with men describe their perceived risk of HIV and what factors influence this risk assessment. We conducted in-depth, virtual interviews with 18 HIV-negative individuals from Philadelphia, eligible for or taking PrEP. The interviews assessed the participants' understanding of their HIV risk, using thematic analysis to deductively code and extract themes. Three themes emerged: (1) participants expressed both deliberative and affective risk perception before and after sexual encounters; (2) participants linked HIV knowledge to risk perception and stigma; (3) participants connected intrinsic and extrinsic factors to risk perception differently. Participants endorsed low overall risk perception, while also describing moments of high affective risk perception after sexual encounters in which they were not able to implement their preferred prevention strategies. Future research should explore helping individuals transform affective risk perception into empowerment around sexual health.


Horizontal gene transfer between Wolbachia and the mosquito Aedes aegypti.
The evolutionary importance of horizontal gene transfer (HGT) from Wolbachia endosymbiotic bacteria to their eukaryotic hosts is a topic of considerable interest and debate. Recent transfers of genome fragments from Wolbachia into insect chromosomes have been reported, but it has been argued that these fragments may be on an evolutionary trajectory to degradation and loss. We have discovered a case of HGT, involving two adjacent genes, between the genomes of Wolbachia and the currently Wolbachia-uninfected mosquito Aedes aegypti, an important human disease vector. The lower level of sequence identity between Wolbachia and insect, the transcription of all the genes involved, and the fact that we have identified homologs of the two genes in another Aedes species (Ae. mascarensis), suggest that these genes are being expressed after an extended evolutionary period since horizontal transfer, and therefore that the transfer has functional significance. The association of these genes with Wolbachia prophage regions also provides a mechanism for the transfer. The data support the argument that HGT between Wolbachia endosymbiotic bacteria and their hosts has produced evolutionary innovation.


Topical corticosteroids and the risk of diabetes mellitus: a nested case-control study in the Netherlands.
The relationship between topical corticosteroid use, potency, treatment duration, concomitant exposure to systemic corticosteroids, and risk of diabetes has been incompletely studied. To investigate an association between intense, longstanding topical corticosteroid use and diabetes mellitus. Data for this nested case-control study were obtained from the PHARMO Record Linkage System, including linked drug dispensing and hospital records of >2.5 million individuals in defined areas of the Netherlands. Users of topical corticosteroids during 1992-2004, without diabetes, with >or=2 topical corticosteroid dispensings and >or=4 years of follow-up were selected. Diabetes onset was defined as first occurrence (index date) of an antidiabetic drug dispensing or hospitalization for diabetes. Cases were matched 1:4 by age and sex to controls, with >or=2 topical corticosteroid dispensings and similar follow-up duration. Use of topical corticosteroids and systemic corticosteroids and/or inhaled corticosteroids as co-medication was classified as current, recent and past/never (<or=2 years, 2-4 years and >4 years ago, respectively). Multivariate regression analyses were adjusted for co-medication and co-morbidity. Among 192,893 incident topical corticosteroid users, 2,212 developed diabetes and could be matched to 8,582 controls. Current topical corticosteroid use was associated with an (unadjusted) 1.24-fold increased risk of diabetes (unadjusted OR 1.24; 95% CI 1.11, 1.40). The odds ratio increased to 1.32 with >180 days of topical corticosteroid use (95% CI 1.14, 1.54) and to 1.44 with a cumulative topical corticosteroid load (combined potency and amount) of 731-1460 mg (95% CI 1.21, 1.72). Among past/never users of systemic corticosteroids and/or inhaled corticosteroids, current use of topical corticosteroids remained associated with a 1.27-fold increased diabetes risk (unadjusted OR 1.27; 95% CI 1.10, 1.47) compared with past users of topical corticosteroids. An increased risk of new-onset diabetes may be an important consideration in the treatment of patients with topical corticosteroids, especially when intense skin treatment is needed. Future studies are needed to endorse these findings in other populations.


Vaccination strategies to prevent emerging diseases for Spanish aquaculture.
In recent years, three serious diseases have emerged in Spanish aquaculture. These are lactococcosis caused by Lactococcus garvieae, which is of economical importance in rainbow trout (Oncorhynchus mykiss); pseudomonadiasis caused by Pseudomonas anguilliseptica which affects gilthead seabream (Sparus aurata) and turbot (Scophthalmus maximus); and flexibacteriosis caused by Tenacibaculum maritimum which became a devastating problem in the emerging culture of sole (Solea spp). To obtain useful information for the design and development of new vaccines, antigenic characterisation of representative strains was performed. In this work we present the strategies adopted for the vaccine formulation (strains included, use of adjuvants) and administration (route, necessity of booster, etc.). The results from laboratory and/or field vaccination trials performed showed that for lactococcosis, protection lasting for five months was obtained with an oil-adjuvanted bacterin formulation. Unadjuvanted bacterin gave only a short duration of protection, which could, however, be prolonged by an antigen boost administered via the feed. A bacterin against Pseudomonas anguilliseptica gave protection for 12 weeks when tested in an experimental challenge trial in turbot. Besides the flexibacteriosis vaccine developed by our group for turbot, and due to the antigenic host-associated variability within T. maritimum, a new bacterin was developed against this bacterium to be used specifically in sole. This new bacterin, administered to sole by intraperitoneal injection, yielded RPS values of 94 % six weeks after immunization. In conclusion, these results suggest that vaccination constitutes a cost-effective method of controlling diseases that have emerged in the most important fish species being cultured in Spain.


Measuring QT dispersion: man versus machine.
To compare manual and computer automated techniques for measuring QT dispersion. Assessment of the ability of manual and automatic measurements of QT dispersion to discriminate between a normal group and two cardiac groups. 12 simultaneous electrocardiogram leads were recorded from 25 healthy volunteers, 25 subjects after myocardial infarction, and 25 with cardiac arrhythmias. For each subject, QT dispersion was measured as the difference between the maximum and minimum QT from all 12 leads and separately for only those leads with T amplitudes of > 100 microV and for those > 250 microV. Manual QT dispersion (T > 100 microV) was greater (P < 0.02) in the arrhythmia patients (mean (SD), 45 (21) ms), but not the infarction patients (54 (36) ms), than in the normal subjects (39 (13) ms). There were no significant differences when all T waves were included. QT dispersion was significantly reduced by an average of 30% when T waves < 100 microV were excluded, and by 51% when those < 250 microV were excluded. Automatic techniques gave different measurements for dispersion in comparison with manual measurements. Three of the four automatic techniques detected significant differences between normal and both patient groups when no leads were excluded (P < 0.01) as well as when T waves < 100 microV were excluded (with increased significance, P < 0.002). Measurements of QT dispersion from small T waves increases measurement variability and reduces the potential for detecting clinical differences. Automatic measurement of QT dispersion gives different results from manual measurement, but can satisfactorily discriminate between normal and abnormal groups with good quality electrocardiograms.


Molecular diagnostics, personalized medicine, and the evolving role of the cytotechnologist: an institutional experience.
The role of cytotechnologists has focused primarily on the microscopic examination of cytologic specimens for diagnosing disease. Cytotechnologists currently evaluate a wide assortment of both gynecological and nongynecological cytology specimens. However, the Pap test remains the primary test for most cytology laboratories. Recently, human papillomavirus testing and newer cervical cancer screening guidelines have reduced the number of Pap tests, resulting in some anxiety and concern among the cytology community. However, as Pap test volumes continue to decrease, molecular oncology and ancillary testing volumes continue to increase with the advent of new biomarkers and associated personalized therapies. This change in clinical practice has resulted in evolving roles for many cytotechnologists. Cytotechnologists have skills based not only in morphology but also in understanding concepts of disease including neoplasia. These skills allow cytotechnologists to excel in many other types of laboratory testing. This article discusses how the roles of the cytotechnologist have recently expanded at our institution to include involvement in DNA ploidy analysis, quantitative immunohistochemistry, fluorescence in situ hybridization, circulating tumor cells, and molecular oncology testing. Lastly, this article discusses how these newer roles benefit both the cytotechnologist and the clinical laboratory.


HOXC10 as a potential marker for discriminating between amnion- and decidua-derived mesenchymal stem cells.
The HOX family of genes plays a fundamental role in the morphogenesis of vertebrate embryonic cells. HOX genes are thought to be important for the regulation of stem cells. We investigated HOX gene expression in mesenchymal stem cells (MSCs) from human placentas. We isolated MSCs from human placentas and confirmed stemness by fluorescence-activated cell sorting (FACS) analysis and differentiation studies. Using reverse transcription PCR, mRNA expression of 39 Class I HOX genes was measured in the MSCs. The expression of HOXB6, C4, C8, C10, D3, D4, and D10 were measured by Western blot analysis. HOXC10 was expressed in 10 of 10 amnion-derived MSCs but in only 2 of 10 decidua-derived MSCs. HOXC4 and D10 were expressed in 100% of both amnion-derived MSCs and deciduas-derived MSCs. HOXD4 was silent in all amnion-derived MSCs and deciduas-derived MSCs (n = 10). HOX gene activation patterns might be a useful indicator for the detection of MSCs of different tissue origins. We demonstrated that HOXC10 is a gene that may discriminate between amnion-derived MSCs and decidua-derived MSCs.


Dynamic stereometry of the temporomandibular joint.
Studies on jaw kinematics have provided a good understanding of the motion of the mandible in space, but are of little biomechanical relevance because they could not relate the movements to anatomic structures. This is possible by the combination of three-dimensional reconstructions of the temporomandibular joint (TMJ) anatomy with jaw motion recordings. This technique allows us to analyze the variation of the relationship between the articular surfaces, providing indirect insight into disk deformation during function and parafunction as well as TMJ loading. As far as the variation of the condyle-fossa distance is concerned, data indicated that during chewing the distance was smaller 1) on closing than on opening; 2) on the balancing than on the working side; and 3) during chewing of hard than soft food. Moreover, during a forceful static biting, the condyle-fossa distance decreased more on the contralateral, i.e. on the balancing side than on the working side. The decrease was related to the degree of clenching force. These results support the content that both condyles are loaded during chewing and the balancing side joint more than the working one. Biomechanically, the development of osteoarthrosis is more likely related to the magnitude and frequency of stresses applied on the cartilage. Joint movements produce tractional forces that may cause shear stresses contributing to cartilage wear and fatigue. Tractional forces are the result of frictional forces caused by the cartilage surface rubbing and of plowing forces caused by the translation of a stress-field through the cartilage matrix, as the intra-articular space changes during motion. Translation of the stress-field in mediolateral direction seems to be particularly important for the integrity of the TMJ disk because of its anisotropic properties. Dynamic stereometry showed that stress-fields translate in mediolateral direction during opening/closing, protrusion and laterotrusion, and that their translatory velocity varies intraindividually and with the rate of the condylar movement. Furthermore, the results seem to indicate that the lateral area of the TMJ disk is more often exposed to shear stresses caused by stress-field translation than the medial one. In conclusion, dynamic stereometry provides a good visualization of the movement of the condyles in the respective fossae. This helps improving our understanding for the complexity of condylar movements. The technique may also contribute to ameliorate our knowledge of TMJ biomechanics and therefore of the etiology of degenerative joint diseases and possibly also of internal derangement.


Integrating 5-hydroxymethylcytosine into the epigenomic landscape of human embryonic stem cells.
Covalent modification of DNA distinguishes cellular identities and is crucial for regulating the pluripotency and differentiation of embryonic stem (ES) cells. The recent demonstration that 5-methylcytosine (5-mC) may be further modified to 5-hydroxymethylcytosine (5-hmC) in ES cells has revealed a novel regulatory paradigm to modulate the epigenetic landscape of pluripotency. To understand the role of 5-hmC in the epigenomic landscape of pluripotent cells, here we profile the genome-wide 5-hmC distribution and correlate it with the genomic profiles of 11 diverse histone modifications and six transcription factors in human ES cells. By integrating genomic 5-hmC signals with maps of histone enrichment, we link particular pluripotency-associated chromatin contexts with 5-hmC. Intriguingly, through additional correlations with defined chromatin signatures at promoter and enhancer subtypes, we show distinct enrichment of 5-hmC at enhancers marked with H3K4me1 and H3K27ac. These results suggest potential role(s) for 5-hmC in the regulation of specific promoters and enhancers. In addition, our results provide a detailed epigenomic map of 5-hmC from which to pursue future functional studies on the diverse regulatory roles associated with 5-hmC.


14-day bactericidal activity of PA-824, bedaquiline, pyrazinamide, and moxifloxacin combinations: a randomised trial.
New drugs, but also shorter, better-tolerated regimens are needed to tackle the high global burden of tuberculosis complicated by drug resistance and retroviral disease. We investigated new multiple-agent combinations over the first 14 days of treatment to assess their suitability for future development. In this prospective, randomised, early bactericidal activity (EBA) study, treatment-naive, drug-susceptible patients with uncomplicated pulmonary tuberculosis were admitted to hospitals in Cape Town, South Africa, between Oct 7, 2010, and Aug 19, 2011. Patients were randomised centrally by computer-generated randomisation sequence to receive bedaquiline, bedaquiline-pyrazinamide, PA-824-pyrazinamide, bedaquiline-PA-824, PA-824-moxifloxacin-pyrazinamide, or unmasked standard antituberculosis treatment as positive control. The primary outcome was the 14-day EBA assessed in a central laboratory from the daily fall in colony forming units (CFU) of M tuberculosis per mL of sputum in daily overnight sputum collections. Bilinear regression curves were fitted for each group separately and groups compared with ANOVA for ranks, followed by pair-wise comparisons adjusted for multiplicity. Clinical staff were partially masked but laboratory personnel were fully masked. This study is registered, NCT01215851. The mean 14-day EBA of PA-824-moxifloxacin-pyrazinamide (n=13; 0·233 [SD 0·128]) was significantly higher than that of bedaquiline (14; 0·061 [0·068]), bedaquiline-pyrazinamide (15; 0·131 [0·102]), bedaquiline-PA-824 (14; 0·114 [0·050]), but not PA-824-pyrazinamide (14; 0·154 [0·040]), and comparable with that of standard treatment (ten; 0·140 [0·094]). Treatments were well tolerated and appeared safe. One patient on PA-824-moxifloxacin-pyrazinamide was withdrawn because of corrected QT interval changes exceeding criteria prespecified in the protocol. PA-824-moxifloxacin-pyrazinamide is potentially suitable for treating drug-sensitive and multidrug-resistant tuberculosis. Multiagent EBA studies can contribute to reducing the time needed to develop new antituberculosis regimens. The Global Alliance for TB Drug Development (TB Alliance).


Towards social inclusion through lifelong learning in mental health: analysis of change in the lives of the EMILIA project service users.
The application of formal lifelong learning to enhance social inclusion in mental health is rarely investigated in terms of change in the lives of service users on a cross-country comparative scale. This study was aimed at examining changes in key areas of the lives of mental health service users across eight European mental health sites. A before and after case study design was applied. Users of mental health services who participated in the lifelong leaning interventions reviewed the changes in key areas of their lives at baseline and 10 months later, through the thematic analysis of qualitative data collected in semi-structured interviews (27 and 21, respectively) and self-reports (138 and 99, respectively). In-depth examples from one site are provided. Most users reported positive changes in the areas of training and social networks, with a sizeable minority moving onto unpaid and paid employment. In addition most users reported active planning for job search and other goals. Obstacles that were highlighted included the negative effects of having a mental illness, difficulties in close relationships and economic disadvantages. The lifelong learning intervention offered within an EU Framework 6 project to mental health service users in eight demonstration sites had a largely positive impact on key areas of their lives at 10 months, though obstacles remained which may be less amenable to change by social interventions.


The Difficulties in Antihypertensive Drug Prescription During Lactation: Is the Information Consistent?
The lack of consistent official information on the use of medications during lactation is probably one of the main reasons leading to an excess of prudence, based on presumption of risk rather than on evidence. The objective of this study was to compare the level of agreement between different official sources available to doctors and women on the use of medications during lactation. Sources of information included governmental regulatory agencies (RAs) and scientific sources (SS). The package leaflets (PLs) and summaries of product characteristics (SPCs) were retrieved from online databases of the European Union and U.S. RAs. Among the SS, the latest edition of the book Medications and Mothers' Milk by Hale and Rowe and the LactMed database were selected. Information about the use of 11 antihypertensive medications during breastfeeding was analyzed. The PLs and SPCs report a higher risk profile than the one expressed by SS, and they often suggest the interruption of lactation even for compatible medications. Health professionals should be supported by official, accurate, comprehensive, and consistent information about maternally ingested medication and breastfeeding management to facilitate proper decision-making.


23. Pain in patients with cancer.
Pain in patients with cancer can be refractory to pharmacological treatment or intolerable side effects of pharmacological treatment may seriously disturb patients' quality of life. Specific interventional pain management techniques can be an effective alternative for those patients. The appropriate application of these interventional techniques provides better pain control, allows the reduction of analgesics and hence improves quality of life. Until recently, the majority of these techniques are considered to be a fourth consecutive step following the World Health Organization's pain treatment ladder. However, in cancer patients, earlier application of interventional pain management techniques can be recommended even before considering the use of strong opioids. Epidural and intrathecal medication administration allow the reduction of the daily oral or transdermal opioid dose, while maintaining or even improving the pain relief and reducing the side effects. Cervical cordotomy may be considered for patients suffering with unilateral pain at the level below the dermatome C5. This technique should only be applied in patients with a life expectancy of less than 1 year. Plexus coeliacus block or nervus splanchnicus block are recommended for the management of upper abdominal pain due to cancer. Pelvic pain due to cancer can be managed with plexus hypogastricus block and the saddle or lower end block may be a last resort for patients suffering with perineal pain. Back pain due to vertebral compression fractures with or without pathological tumor invasion may be managed with percutaneous vertebroplasty or kyphoplasty. All these interventional techniques should be a part of multidisciplinary patient program.


Whole-Transcriptome Amplification of Single Cells for Next-Generation Sequencing.
Single-cell transcriptome analysis gives insight into the heterogeneity of neighboring cells in tissues or cell cultures. It has been shown that the variability of cells is important for biological function. However, cell variability can be an indication of disease (e.g., cancer). In order to elucidate cell variability in health and disease, single-cell transcriptomes can be analyzed by new next-generation sequencing (NGS) platforms. NGS analysis currently requires a few hundred nanograms of RNA. Consequently, accurate whole-transcriptome amplification is required to analyze the transcriptome of a single cell by NGS. This unit describes the entire workflow from cells to NGS, including cell quality testing, cell lysis, gDNA removal, whole-transcriptome amplification, and NGS library preparation. Recommendations for WTA quality control are given and optional protocols for using purified RNA for WTA and WTA purification are described.


Nitric oxide signaling mechanisms in the forearm vasculature of pregnant women.
During pregnancy, forearm blood flow and constrictor responses to nitric oxide synthase inhibition are enhanced. Responses to brachial artery infusion of N(G)-monomethyl-L -arginine in pregnant women who have normal resting forearm blood flow and in pregnant women who have norepinephrine-induced reduced forearm blood flow were no different. Mechanisms independent of acute flow effects may account for the enhanced vascular nitric oxide activity during pregnancy.