[Primary Healthcare Reform in Portugal on two fronts: autonomous family healthcare units and management of groupings of Health Centers].
In 2005, Portugal began a reform of Primary Health Care. This reform process through to April 2010 is described and analyzed. During this period the Mission for Primary Health Care was responsible for conducting a profound reconfiguration. The main objectives for this reform were to improve accessibility, efficiency, quality and continuity of care and increase the satisfaction of professionals and citizens. The main features are voluntary adhesion, teamwork, mandatory information system, performance-sensitive payment, contracting and evaluation. The reconfiguration of health centers was two pronged. First, there was the formation of small autonomous functional units, known as Family Health Units (USF) providing services with proximity and quality. The second measure involved the aggregation of resources and management structures, groups of health centers (ACES), seeking to achieve efficiency and economies of scale. The FHU proved to offer simultaneously more efficiency, accessibility, better working environment, greater citizen satisfaction, namely better quality. The importance of strong political support, the creation of a structure responsible for the design and implementation of reform and good liaison with the media are stressed.


A spring forward for hominin evolution in East Africa.
Groundwater is essential to modern human survival during drought periods. There is also growing geological evidence of springs associated with stone tools and hominin fossils in the East African Rift System (EARS) during a critical period for hominin evolution (from 1.8 Ma). However it is not known how vulnerable these springs may have been to climate variability and whether groundwater availability may have played a part in human evolution. Recent interdisciplinary research at Olduvai Gorge, Tanzania, has documented climate fluctuations attributable to astronomic forcing and the presence of paleosprings directly associated with archaeological sites. Using palaeogeological reconstruction and groundwater modelling of the Olduvai Gorge paleo-catchment, we show how spring discharge was likely linked to East African climate variability of annual to Milankovitch cycle timescales. Under decadal to centennial timescales, spring flow would have been relatively invariant providing good water resource resilience through long droughts. For multi-millennial periods, modelled spring flows lag groundwater recharge by 100 s to 1000 years. The lag creates long buffer periods allowing hominins to adapt to new habitats as potable surface water from rivers or lakes became increasingly scarce. Localised groundwater systems are likely to have been widespread within the EARS providing refugia and intense competition during dry periods, thus being an important factor in natural selection and evolution, as well as a vital resource during hominin dispersal within and out of Africa.


Tai chi qigong as a means to improve night-time sleep quality among older adults with cognitive impairment: a pilot randomized controlled trial.
Age-related cognitivee decline is a growing public health concern worldwide. More than a quarter of adults with cognitive impairment experience sleep disturbance. The objective of this pilot study was to evaluate the preliminary effects of tai chi qigong (TCQ) on improving the night-time sleep quality of older adults with cognitive impairment. Older adults with cognitive impairment who complain of sleep disturbance. A randomized controlled trial with two groups. Fifty-two subjects were recruited from two district elderly community centers and randomly assigned to either the TCQ group (n=27) or the control group (n=25). The intervention group received TCQ training consisting of two 60-minute sessions each week for 2 months. The control group was advised to maintain their usual activities. Sleep quality was measured by the Chinese Pittsburgh Sleep Quality Index. Quality of life was measured by Short-form 12, cognitive functions measured by mini-mental state examination, and subjective memory deficits measured by the memory inventory for Chinese. Data were collected at baseline, 2 months, and 6 months. Significant results were noted at 6 months in the Chinese Pittsburgh Sleep Quality Index global score (P=0.004), sleep duration (P=0.003), habitual sleep efficiency (P=0.002), and the Short-form 12 mental health component (P<0.001). The TCQ participants reported better sleep quality and a better (quality of life) mental health component than the control group. TCQ can be considered a useful nonpharmacological approach for improving sleep quality in older adults with cognitive impairment. CUHK_CCT00448 (https://www2.ccrb.cuhk.edu.hk/registry/public/287).


Are brief interventions to increase physical activity cost-effective? A systematic review.
To determine whether brief interventions promoting physical activity are cost-effective in primary care or community settings. Systematic review of economic evaluations. We searched MEDLINE, EMBASE, PsycINFO, CINAHL, EconLit, SPORTDiscus, PEDro, the Cochrane library, National Health Service Economic Evaluation Database and the Cost-Effectiveness Analysis Registry up to 20 August 2014. Web of Knowledge was used for cross-reference search. We included studies investigating the cost-effectiveness of brief interventions, as defined by National Institute for Health and Care Excellence, promoting physical activity in primary care or the community. Methodological quality was assessed using Drummond's checklist for economic evaluations. Data were extracted from individual studies fulfilling selection criteria using a standardised pro forma. Comparisons of cost-effectiveness and cost-utility ratios were made between studies. Of 1840 identified publications, 13 studies fulfilled the inclusion criteria describing 14 brief interventions. Studies varied widely in the methods used, such as the perspective of economic analysis, intervention effects and outcome measures. The incremental cost of moving an inactive person to an active state, estimated for eight studies, ranged from £96 to £986. The cost-utility was estimated in nine studies compared with usual care and varied from £57 to £14 002 per quality-adjusted life year; dominant to £6500 per disability-adjusted life year; and £15 873 per life years gained. Brief interventions promoting physical activity in primary care and the community are likely to be inexpensive compared with usual care. Given the commonly accepted thresholds, they appear to be cost-effective on the whole, although there is notable variation between studies.


The differential pupillary response to 2.5% phenylephrine in patients taking tamsulosin.
To determine if a pharmacological test could be developed to determine iris dilator dysfunction in patients taking tamsulosin. Patients taking tamsulosin and controls were recruited from the Urology and Ophthalmology clinics at the Montefiore Medical Center. The patient's right eye (OD) was dilated with phenylephrine hydrochloride 2.5% and tropicamide 1%. The patient's left eye (OS) was dilated with tropicamide 1% alone. Forty minutes after dilation, pupillary diameter was measured in both eyes. Thirty-eight tamsulosin subjects and 43 controls met the inclusion criteria for the study. The tamsulosin-treated patients dilated less with phenylephrine than controls (0.61±0.4 vs. 1.10±0.45 mm, respectively P<0.001). Receiver operating characteristic curves comparing maximal pupillary dilation versus differential pupillary dilation in tamsulosin patients relative to controls shows a greater area under the curve for differential dilation (0.8 vs. 0.6, respectively). A correlation between smooth muscle dysfunction and length of time on tamsulosin was observed. Patients using tamsulosin for <1 month had an average OD-OS difference of 0.85±0.5 mm. Patients who were on tamsulosin for >1 month had an average OD-OS difference of 0.52±0.32 mm (P<0.01, Mann-Whitney). Patients treated with tamsulosin demonstrated a significantly decreased iris dilatory response to the selective adrenergic effects of phenylephrine compared to controls. Additionally, it appears that longer duration of exposure to tamsulosin increases the likelihood of dilator dysfunction.


Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes.
We compared commonly used BAPTA-derived chemical Ca2+ dyes (fura2, Fluo-4, and Rhod-2) with a newer genetically encoded indicator (R-GECO) in single cell models of the heart. We assessed their performance and effects on cardiomyocyte contractility, determining fluorescent signal-to-noise ratios and sarcomere shortening in primary ventricular myocytes from adult mouse and guinea pig, and in human iPSC-derived cardiomyocytes. Chemical Ca2+ dyes displayed dose-dependent contractile impairment in all cell types, and we observed a negative correlation between contraction and fluorescence signal-to-noise ratio, particularly for fura2 and Fluo-4. R-GECO had no effect on sarcomere shortening. BAPTA-based dyes, but not R-GECO, inhibited in vitro acto-myosin ATPase activity. The presence of fura2 accentuated or diminished changes in contractility and Ca2+ handling caused by small molecule modulators of contractility and intracellular ionic homeostasis (mavacamten, levosimendan, and flecainide), but this was not observed when using R-GECO in adult guinea pig left ventricular cardiomyocytes. Ca2+ handling studies are necessary for cardiotoxicity assessments of small molecules intended for clinical use. Caution should be exercised when interpreting small molecule studies assessing contractile effects and Ca2+ transients derived from BAPTA-like chemical Ca2+ dyes in cellular assays, a common platform for cardiac toxicology testing and mechanistic investigation of cardiac disease physiology and treatment.


Regulation of Excitatory Amino Acid Transmission in the Retina: Studies on Neuroprotection.
Excitotoxicity occurs in neurons due to the accumulation of excitatory amino acids such as glutamate in the synaptic and extrasynaptic locations. In the retina, excessive glutamate concentrations trigger a neurotoxic cascade involving several mechanisms, including the elevation of intracellular calcium (Ca2+) and the activation of α-amino-3-hydroxy 5-methyl-4-iso-xazole-propionic acid/kainate (AMPA/KA) and N-methyl-d-aspartate (NMDA) receptors leading to retinal degeneration. Both ionotropic glutamate receptors (iGluRs) and metabotropic glutamate receptors (mGluRs) are present in the mammalian retina. Indeed, due to the abundant expression of GluRs, the mammalian retina is highly susceptible to excitotoxic neurodegeneration. Excitotoxicity has been postulated to present a common downstream mechanism for several stimuli, including hypoglycemia, hypoxia, ischemia, and chronic neurodegenerative diseases. Experimental approaches to the study of neuroprotection in the retina have utilized insults that trigger hypoxia, hypoglycemia, or excitotoxicity. Using these experimental approaches, the neuroprotective potential of GluR agents, including the NMDA receptor modulators (MK801, ifenprodil, memantine); AMPA/KA receptor antagonist (CNQX); Group II and III mGluR agonists (LY354740, quisqualate); and Ca2+-channel blockers (diltiazem, lomerizine, verapamil, ω-conotoxin), and others (pituitary adenylate cyclase activating polypeptide, neuropeptide Y, acetylcholine receptor agonists) have been elucidated. In addition to corroborating the exocytotic role of excitatory amino acids in retinal degeneration, these studies affirm that multiple mechanism/s contribute to the prevention of damage caused by excitotoxicity in the retina. Therefore, it is feasible that several pathways are involved in protecting the retina from toxic insults in ocular neurodegenerative conditions such as glaucoma and retinal ischemia. Furthermore, these experimental models are viable tools for evaluating therapeutic candidates in ocular neuropathies.


Serum Pentraxin 3 is associated with signs of arterial alteration in women with preeclampsia.
Preeclampsia (PE) in pregnancy is a state of exaggerated inflammation and is associated with an increased risk of cardiovascular disease (CVD) later in life. Levels of pentraxin 3 (PTX3), a novel inflammation marker, are increased during PE and in individuals with CVD. The primary aim of this study was to assess whether serum PTX3 in women with PE is associated with adverse arterial effects; a thicker intima and higher intima/media (I/M) ratio in the common carotid artery (CCA). Serum PTX3 levels were measured using commercially available enzyme-linked immunosorbent assay kits, and individual CCA intima and media thicknesses were estimated by 22MHz non-invasive ultrasound in 55 women at PE diagnosis and 64 women with normal pregnancies at a similar gestational age, and about one year postpartum. A thick intima, thin media and high I/M ratio indicate a less healthy artery wall. During pregnancy serum PTX3 correlated positively with intima thickness and I/M ratio but negatively with media thickness (all p<0.0001), indicating adverse arterial effects. About one year postpartum, PTX3 levels had decreased in both groups and there remained no significant group difference or significant correlation with CCA wall layers. Higher levels of serum PTX3 in women with PE were significantly associated with signs of adverse arterial effects during pregnancy, but not one year postpartum, supporting the rapid dynamics of PTX3.


Interaction between mutant ataxin-1 and PQBP-1 affects transcription and cell death.
PQBP-1 was isolated on the basis of its interaction with polyglutamine tracts. In this study, using in vitro and in vivo assays, we show that the association between ataxin-1 and PQBP-1 is positively influenced by expanded polyglutamine sequences. In cell lines, interaction between the two molecules induces apoptotic cell death. As a possible mechanism underlying this phenomenon, we found that mutant ataxin-1 enhances binding of PQBP-1 to the C-terminal domain of RNA polymerase II large subunit (Pol II). This reduces the level of phosphorylated Pol II and transcription. Our results suggest the involvement of PQBP-1 in the pathology of spinocerebellar ataxia type 1 (SCA1) and support the idea that modified transcription underlies polyglutamine-mediated pathology.


Mapping between the Roland Morris Questionnaire and generic preference-based measures.
The Roland Morris Questionnaire (RMQ) is a widely used health status measure for low back pain (LBP). It is not preference-based, and there are currently no established algorithms for mapping between the RMQ and preference-based health-related quality-of-life measures. Using data from randomized controlled trials of treatment for LBP, we sought to develop algorithms for mapping between RMQ scores and health utilities derived using either the EuroQol five-dimensional questionnaire (EQ-5D) or the six-dimensional health state short form (derived from Medical Outcomes Study 36-Item Short-Form Health Survey) (SF-6D). This study is based on data from the Back Skills Training Trial in which data were collected from 701 patients at baseline and subsequently at 3, 6, and 12 months postrandomization using a range of outcome measures, including the RMQ, the EQ-5D, and the Short Form 12 item Health Survey (SF-12) (from which SF-6D utilities can be derived). We used baseline trial data to estimate models using both direct and response mapping approaches to predict EQ-5D and SF-6D health utilities and dimension responses. A multistage model selection process was used to assess the predictive accuracy of the models. We then explored different techniques and mapping models that made use of repeated follow-up observations in the data. The estimated mapping algorithms were validated using external data from the UK Back Pain Exercise and Manipulation trial. A number of models were developed that accurately predict health utilities in this context. The best performing model for RMQ to EQ-5D mapping was a beta regression with Bayesian quasi-likelihood estimation that included 24 dummy variables for RMQ responses, age, and sex as covariates (mean squared error 0.0380) based on repeated data. The model selected for RMQ to SF-6D mapping was a finite mixture model that included the overall RMQ score, age, sex, RMQ score squared, age squared, and an interaction term for age and RMQ score as covariates (mean squared error 0.0114) based on repeated data. It is possible to reasonably predict EQ-5D and SF-6D health utilities from RMQ scores and responses using regression methods. Our regression equations provide an empirical basis for estimating health utilities when EQ-5D or SF-6D data are not available. They can be used to inform future economic evaluations of interventions targeting LBP.


Investigation on the Power Factor of Skutterudite Smy(FexNi1-x)4Sb12 Thin Films: Effects of Deposition and Annealing Temperature.
Filled skutterudites are currently studied as promising thermoelectric materials due to their high power factor and low thermal conductivity. The latter property, in particular, can be enhanced by adding scattering centers, such as the ones deriving from low dimensionality and the presence of interfaces. This work reports on the synthesis and characterization of thin films belonging to the Smy(FexNi1-x)4Sb12-filled skutterudite system. Films were deposited under vacuum conditions by the pulsed laser deposition (PLD) method on fused silica substrates, and the deposition temperature was varied. The effect of the annealing process was studied by subjecting a set of films to a thermal treatment for 1 h at 423 K. Electrical conductivity σ and Seebeck coefficient S were acquired by the four-probe method using a ZEM-3 apparatus performing cycles in the 348-523 K temperature range, recording both heating and cooling processes. Films deposited at room temperature required three cycles up to 523 K before being stabilized, thus revealing the importance of a proper annealing process in order to obtain reliable physical data. XRD analyses confirm the previous result, as only annealed films present a highly crystalline skutterudite not accompanied by extra phases. The power factor of annealed films is shown to be lower than in the corresponding bulk samples due to the lower Seebeck coefficients occurring in films. Room temperature thermal conductivity, on the contrary, shows values comparable to the ones of doubly doped bulk samples, thus highlighting the positive effect of interfaces on the introduction of scattering centers, and therefore on the reduction of thermal conductivity.


Smoking among Nepali youth--prevalence and predictors.
Tobacco is the single largest risk factor for various diseases and its presence in the young heralds more serious problems as they may be exposed for longer periods. Prevalence and predictors of smoking among youth will aid in formulating effective preventive and control measures. A cross-sectional study was therefore conducted among 816 students selected from five colleges of Western Nepal using a self-administered questionnaire. Prevalence of ever smoking was 34.2% (males 47.6% and females 18.4%) and for current smoking was 17%. It was higher among youth belonging to 21 years or older as compared to younger age groups. Mean age of initiation was 16.8 years (standard deviation 2.8 years) and the most common reasons cited for smoking were; like it, to feel more relaxed, out of boredom and to look more mature. Proportion of youth who said they felt they were addicted was 43.1% and 64.7% said that they had tried to quit the habit. Most important predictors having independent effects on youth being ever smokers were having three or more smoker friends (OR=18), their own chewing (OR=4.8) or alcohol use (OR=4.2), male gender (OR=3.7) and the type of course they were pursuing, with professional course students having higher risk. With almost one fifth of college-going youth smoking and a higher prevalence in older age groups within them, smoking is a serious concern for young people in Western Nepal.


Quality of scientific articles.
The paper discusses the difficulties in judging the quality of scientific manuscripts and describes some common pitfalls that should be avoided when preparing a paper for submission to a peer-reviewed journal. Peer review is an imperfect system, with less than optimal reliability and uncertain validity. However, as it is likely that it will remain as the principal process of screening papers for publication, authors should avoid some common mistakes when preparing a report based on empirical findings of human research. Among these are: excessively long abstracts, extensive use of abbreviations, failure to report results of parsimonious data analyses, and misinterpretation of statistical associations identified in observational studies as causal. Another common problem in many manuscripts is their excessive length, which makes them more difficult to be evaluated or read by the intended readers, if published. The evaluation of papers after their publication with a view towards their inclusion in a systematic review is also discussed. The limitations of the impact factor as a criterion to judge the quality of a paper are reviewed.


Patient and caregiver perspectives on early identification for advance care planning in primary healthcare settings.
As part of a broader study to improve the capacity for advance care planning (ACP) in primary healthcare settings, the research team set out to develop and validate a computerized algorithm to help primary care physicians identify individuals at risk of death, and also carried out focus groups and interviews with relevant stakeholder groups. Interviews with patients and family caregivers were carried out in parallel to algorithm development and validation to examine (1) views on early identification of individuals at risk of deteriorating health or dying; (2) views on the use of a computerized algorithm for early identification; and (3) preferences and challenges for ACP. Fourteen participants were recruited from two Canadian provinces. Participants included individuals aged 65 and older with declining health and self-identified caregivers of individuals aged 65 and older with declining health. Semi-structured interviews were conducted via telephone. A qualitative descriptive analytic approach was employed, which focused on summarizing and describing the informational contents of the data. Participants supported the early identification of patients at risk of deteriorating health or dying. Early identification was viewed as conducive to planning not only for death, but for the remainder of life. Participants were also supportive of the use of a computerized algorithm to assist with early identification, although limitations were recognized. While participants felt that having family physicians assume responsibility for early identification and ACP was appropriate, questions arose around feasibility, including whether family physicians have sufficient time for ACP. Preferences related to the content of and approach to ACP discussions were highly individualized. Required supports during ACP include informational and emotional supports. This work supports the role of primary care providers in the early identification of individuals at risk of deteriorating health or death and the process of ACP. To improve ACP capacity in primary healthcare settings, compensation systems for primary care providers should be adjusted to ensure appropriate compensation and to accommodate longer ACP appointments. Additional resources and more established links to community organizations and services will also be required to facilitate referrals to relevant community services as part of the ACP process.


Molecular phylogeography of a human autosomal skin color locus under natural selection.
Divergent natural selection caused by differences in solar exposure has resulted in distinctive variations in skin color between human populations. The derived light skin color allele of the SLC24A5 gene, A111T, predominates in populations of Western Eurasian ancestry. To gain insight into when and where this mutation arose, we defined common haplotypes in the genomic region around SLC24A5 across diverse human populations and deduced phylogenetic relationships between them. Virtually all chromosomes carrying the A111T allele share a single 78-kb haplotype that we call C11, indicating that all instances of this mutation in human populations share a common origin. The C11 haplotype was most likely created by a crossover between two haplotypes, followed by the A111T mutation. The two parental precursor haplotypes are found from East Asia to the Americas but are nearly absent in Africa. The distributions of C11 and its parental haplotypes make it most likely that these two last steps occurred between the Middle East and the Indian subcontinent, with the A111T mutation occurring after the split between the ancestors of Europeans and East Asians.


Melittin restores proteasome function in an animal model of ALS.
Amyotrophic lateral sclerosis (ALS) is a paralyzing disorder characterized by the progressive degeneration and death of motor neurons and occurs both as a sporadic and familial disease. Mutant SOD1 (mtSOD1) in motor neurons induces vulnerability to the disease through protein misfolding, mitochondrial dysfunction, oxidative damage, cytoskeletal abnormalities, defective axonal transport- and growth factor signaling, excitotoxicity, and neuro-inflammation.Melittin is a 26 amino acid protein and is one of the components of bee venom which is used in traditional Chinese medicine to inhibit of cancer cell proliferation and is known to have anti-inflammatory and anti-arthritic effects.The purpose of the present study was to determine if melittin could suppress motor neuron loss and protein misfolding in the hSOD1G93A mouse, which is commonly used as a model for inherited ALS. Meltittin was injected at the 'ZuSanLi' (ST36) acupuncture point in the hSOD1G93A animal model. Melittin-treated animals showed a decrease in the number of microglia and in the expression level of phospho-p38 in the spinal cord and brainstem. Interestingly, melittin treatment in symptomatic ALS animals improved motor function and reduced the level of neuron death in the spinal cord when compared to the control group. Furthermore, we found increased of α-synuclein modifications, such as phosphorylation or nitration, in both the brainstem and spinal cord in hSOD1G93A mice. However, melittin treatment reduced α-synuclein misfolding and restored the proteasomal activity in the brainstem and spinal cord of symptomatic hSOD1G93A transgenic mice.Our research suggests a potential functional link between melittin and the inhibition of neuroinflammation in an ALS animal model.


Vitamin C promotes widespread yet specific DNA demethylation of the epigenome in human embryonic stem cells.
Vitamin C (ascorbate) is a widely used medium supplement in embryonic stem cell culture. Here, we show that ascorbate causes widespread, consistent, and remarkably specific DNA demethylation of 1,847 genes in human embryonic stem cells (hESCs), including important stem cell genes, with a clear bias toward demethylation at CpG island boundaries. We show that a subset of these DNA demethylated genes displays concomitant gene expression changes and that the position of the demethylated CpGs relative to the transcription start site is correlated to such changes. We further show that the ascorbate-demethylated gene set not only overlaps with gene sets that have bivalent marks, but also with the gene sets that are demethylated during differentiation of hESCs and during reprogramming of fibroblasts to induced pluritotent stem cells (iPSCs). Our data thus identify a novel link between ascorbate-mediated signaling and specific epigenetic changes in hESCs that might impact on pluripotency and reprogramming pathways.


Block copolymer-quantum dot micelles for multienzyme colocalization.
To mimic the structure and functionality of multienzyme complexes, which are widely present in Nature, Pluronic-based micelles were designed to colocalize multiple enzymes. To stabilize the micelles as well as to enable characterization of single enzyme immobilization and multienzyme colocalization by Förster resonance energy transfer (FRET), quantum dots (QDs) were incorporated into the micelles to form Pluronic-QD micelles using a novel microreactor. Model enzymes glucose oxidase (GOX) and horseradish peroxidase (HRP) were respectively labeled with fluorescent dyes. The results indicated that FRET occurred between the QDs and dyes that labeled each type of enzyme in single enzyme immobilization studies as well as between the dyes in colocalization studies. These observations were consistent with increases in micelle size after adsorption of dye-enzymes as verified by dynamic light scattering. In addition, the activity of single enzymes was retained after immobilization. An optimized colocalization process improved the overall conversion rate by approximately 100% compared to equivalent concentrations of free enzymes in solution. This study demonstrates a versatile platform for multienzyme colocalization and an effective strategy to characterize multienzyme immobilization and colocalization, which can be applicable to many other multienzyme systems.


Identification of tubulin isoforms in the plasmodium of Physarum polycephalum by in vitro microtubule assembly.
The tubulins of the plasmodium of Physarum polycephalum have been identified by in vitro microtubule assembly from partially purified extracts of asynchronous microplasmodia and late G2 macroplasmodia. The plasmodial tubulin group comprised of 2 alpha tubulins (app. m.w. 51000 daltons) and 2 beta tubulins (app. m.w. 58000 daltons and 55000 daltons) and appeared to be identical with a group of polypeptides which are synthesized periodically in late G2. Two of the plasmodial tubulin subunits (one alpha and one beta) were identical to the Physarum amoebal tubulin alpha and beta subunits as characterised by 2D gel positions.


The Lewis superacid Al[N(C6F5)2]3 and its higher homolog Ga[N(C6F5)2]3 - structural features, theoretical investigation and reactions of a metal amide with higher fluoride ion affinity than SbF5.
Herein we present the synthesis of the two Lewis acids Al[N(C6F5)2]3 (ALTA) and Ga[N(C6F5)2]3 (GATA) via salt elimination reactions. The metal complexes were characterized by NMR-spectroscopic methods and X-ray diffraction analysis revealing the stabilization of the highly Lewis acidic metal centers by secondary metal-fluorine contacts. The Lewis acidic properties of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 are demonstrated by reactions with Lewis bases resulting in the formation of metallates accompanied by crucial structural changes. The two metallates [Cs(Tol)3]+[FAl(N(C6F5)2)3]- and [AsPh4]+[ClGa(N(C6F5)2)3]- contain interesting weakly coordinating anions. The reaction of Al[N(C6F5)2]3 with trityl fluoride yielded [CPh3]+[FAl(N(C6F5)2)3]- which could find application in the activation of metallocene polymerization catalysts. The qualitative Lewis acidity of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 was investigated by means of competition experiments for chloride ions in solution. DFT calculations yielded fluoride ion affinities in the gas phase (FIA) of 555 kJ mol-1 for Al[N(C6F5)2]3 and 472 kJ mol-1 for Ga[N(C6F5)2]3. Thus, Al[N(C6F5)2]3 can be considered a Lewis superacid with a fluoride affinity higher than SbF5 (493 kJ mol-1) whereas the FIA of the corresponding gallium complex is slightly below the threshold to Lewis superacidity.


[A retrospective study analysis of urinary hippuric acid levels in occupational toxicology exams].
Hippuric acid is the primary metabolite of toluene, a solvent widely used in industrial processes with considerable toxic effects, a fact which justifies regularly monitoring individuals with occupational exposure to this solvent. This work aims at evaluating urinary hippuric acid levels found in workers subject to biological monitoring. A retrospective study was carried out with data referring from 2002 to 2005, in which exams results and employment status were analyzed (periodic, post-employment, and pre-employment exams). Results indicate a significant reduction in hippuric acid levels for 2005. Periodic exams presented higher results than pre-employment and post-employment exams. No significant difference was found in individuals grouped according to their status in each of the established intervals, their reference numbers, and maximum biological levels allowed. Hippuric acid levels detected indicate low risk of toluene exposure for the population under evaluation, probably due to a growing concern with the deployment of measures regarding occupational hygiene.


Screening for Inhibitors of Acetaldehyde Dehydrogenase (AdhE) from Enterohemorrhagic Escherichia coli (EHEC).
Acetaldehyde dehydrogenase (AdhE) is a bifunctional acetaldehyde-coenzyme A (CoA) dehydrogenase and alcohol dehydrogenase involved in anaerobic metabolism in gram-negative bacteria. This enzyme was recently found to be a key regulator of the type three secretion (T3S) system in Escherichia coli. AdhE inhibitors can be used as tools to study bacterial virulence and a starting point for discovery of novel antibacterial agents. We developed a robust enzymatic assay, based on the acetaldehyde-CoA dehydrogenase activity of AdhE using both absorption and fluorescence detection models (Z' > 0.7). This assay was used to screen ~11,000 small molecules in 384-well format that resulted in three hits that were confirmed by resynthesis and validation. All three compounds are noncompetitive with respect to acetaldehyde and display a clear dose-response effect with hill slopes of 1-2. These new inhibitors will be used as chemical tools to study the interplay between metabolism and virulence and the role of AdhE in T3S regulation in gram-negative bacteria, and as starting points for the development of novel antibacterial agents.


Phonics training for English-speaking poor readers.
Around 5% of English speakers have a significant problem with learning to read words. Poor word readers are often trained to use letter-sound rules to improve their reading skills. This training is commonly called phonics. Well over 100 studies have administered some form of phonics training to poor word readers. However, there are surprisingly few systematic reviews or meta-analyses of these studies. The most well-known review was done by the National Reading Panel (Ehri 2001) 12 years ago and needs updating. The most recent review (Suggate 2010) focused solely on children and did not include unpublished studies. The primary aim of this review was to measure the effect that phonics training has on the literacy skills of English-speaking children, adolescents, and adults whose reading was at least one standard deviation (SD), one year, or one grade below the expected level, despite no reported problems that could explain their impaired ability to learn to read. A secondary objective was to explore the impact of various factors, such as length of training or training group size, that might moderate the effect of phonics training on poor word reading skills. We searched the following databases in July 2012: CENTRAL 2012 (Issue 6), MEDLINE 1948 to June week 3 2012, EMBASE 1980 to 2012 week 26, DARE 2013 (Issue 6), ERIC (1966 to current), PsycINFO (1806 to current), CINAHL (1938 to current), Science Citation Index (1970 to 29 June 2012), Social Science Citation Index (1970 to 29 June 2012), Conference Proceedings Citation Index - Science (1990 to 29 June 2012), Conference Proceedings Citation Index - Social Science & Humanities (1990 to 29 June 2012), ZETOC, Index to Theses-UK and Ireland, ClinicalTrials.gov, ICTRP, the metaRegister of Controlled Trials, ProQuest Dissertations and Theses, DART Europe E-theses Portal, Australasian Digital Theses Program, Education Research Theses, Electronic Theses Online System, Networked Digital Library of Theses and Dissertations. Theses Canada portal, www.dissertation.com, and www.thesisabstracts.com. We also contacted experts and examined the reference lists of published studies. We included studies that use randomisation, quasi-randomisation, or minimisation to allocate participants to either a phonics intervention group (phonics alone, phonics and phoneme awareness training, or phonics and irregular word reading training) or a control group (no training or alternative training, such as maths). Participants were English-speaking children, adolescents, or adults whose word reading was below the level expected for their age for no known reason (that is, they had adequate attention and no known physical, neurological, or psychological problems). Two review authors independently selected studies, assessed risk of bias, and extracted data. We found 11 studies that met the criteria for this review. They involved 736 participants. We measured the effect of phonics training on eight outcomes. The amount of evidence for each outcome varied considerably, ranging from 10 studies for word reading accuracy to one study for nonword reading fluency. The effect sizes for the outcomes were: word reading accuracy standardised mean difference (SMD) 0.47 (95% confidence interval (CI) 0.06 to 0.88; 10 studies), nonword reading accuracy SMD 0.76 (95% CI 0.25 to 1.27; eight studies), word reading fluency SMD -0.51 (95% CI -1.14 to 0.13; two studies), reading comprehension SMD 0.14 (95% CI -0.46 to 0.74; three studies), spelling SMD 0.36 (95% CI -0.27 to 1.00; two studies), letter-sound knowledge SMD 0.35 (95% CI 0.04 to 0.65; three studies), and phonological output SMD 0.38 (95% -0.04 to 0.80; four studies). There was one result in a negative direction for nonword reading fluency SMD 0.38 (95% CI -0.55 to 1.32; one study), though this was not statistically significant.We did five subgroup analyses on two outcomes that had sufficient data (word reading accuracy and nonword reading accuracy). The efficacy of phonics training was not moderated significantly by training type (phonics alone versus phonics and phoneme awareness versus phonics and irregular word training), training intensity (less than two hours per week versus at least two hours per week), training duration (less than three months versus at least three months), training group size (one-on-one versus small group training), or training administrator (human administration versus computer administration). Phonics training appears to be effective for improving some reading skills. Specifically, statistically significant effects were found for nonword reading accuracy (large effect), word reading accuracy (moderate effect), and letter-sound knowledge (small-to-moderate effect). For several other outcomes, there were small or moderate effect sizes that did not reach statistical significance but may be meaningful: word reading fluency, spelling, phonological output, and reading comprehension. The effect for nonword reading fluency, which was measured in only one study, was in a negative direction, but this was not statistically significant.Future studies of phonics training need to improve the reporting of procedures used for random sequence generation, allocation concealment, and blinding of participants, personnel, and outcome assessment.


Differential targeting of IL-2 and T cell receptor signaling pathways selectively expands regulatory T cells while inhibiting conventional T cells.
Strategies to expand regulatory T cells hold therapeutic potential for ameliorating T cell-mediated autoimmunity. Recently, we reported that the requirements for T cell receptor signaling in conventional T cell and regulatory T cell proliferation are different. Using mutant mice that display defective T cell receptor-mediated phospholipase Cγ (PLCγ) activation, we hereby demonstrate that PLCγ activation is required for antigen-specific conventional T cell proliferation but not for IL-2-induced regulatory T cell proliferation. This led us to hypothesize that in conjunction with IL-2, pharmacological inhibition of T cell receptor-mediated PLCγ activation might offer a novel therapeutic strategy to expand regulatory T cells while simultaneously inhibiting conventional T cell proliferation. Indeed, using the calcineurin inhibitor Cyclosporine A to inhibit signaling downstream of PLCγ, we found that Cyclosporine A attenuated antigen-specific Tconv proliferation but permitted IL-2-induced regulatory T cell expansion in vitro and in vivo. Furthermore, the combination of Cyclosporine A and IL-2 was superior over either Cyclosporine A or IL-2 monotherapy in protection against the T cell-mediated demyelinating autoimmune disease mouse model, experimental autoimmune encephalomyelitis. Thus, a combination of TCR signaling inhibition and IL-2 might be a beneficial strategy in expanding regulatory T cells and inhibiting conventional T cell proliferation in autoimmune settings.


Unravelling the GSK3β-related genotypic interaction network influencing hippocampal volume in recurrent major depressive disorder.
Glycogen synthase kinase 3β (GSK3β) has been implicated in mood disorders. We previously reported associations between a GSK3β polymorphism and hippocampal volume in major depressive disorder (MDD). We then reported similar associations for a subset of GSK3β-regulated genes. We now investigate an algorithm-derived comprehensive list of genes encoding proteins that directly interact with GSK3β to identify a genotypic network influencing hippocampal volume in MDD. We used discovery (N=141) and replication (N=77) recurrent MDD samples. Our gene list was generated from the NetworKIN database. Hippocampal measures were derived using an optimized Freesurfer protocol. We identified interacting single nucleotide polymorphisms using the machine learning algorithm Random Forest and verified interactions using likelihood ratio tests between nested linear regression models. The discovery sample showed multiple two-single nucleotide polymorphism interactions with hippocampal volume. The replication sample showed a replicable interaction (likelihood ratio test: P=0.0088, replication sample; P=0.017, discovery sample; Stouffer's combined P=0.0007) between genes associated previously with endoplasmic reticulum stress, calcium regulation and histone modifications. Our results provide genetic evidence supporting associations between hippocampal volume and MDD, which may reflect underlying cellular stress responses. Our study provides evidence of biological mechanisms that should be further explored in the search for disease-modifying therapeutic targets for depression.


AMPA receptors as a molecular target in epilepsy therapy.
Epileptic seizures occur as a result of episodic abnormal synchronous discharges in cerebral neuronal networks. Although a variety of non-conventional mechanisms may play a role in epileptic synchronization, cascading excitation within networks of synaptically connected excitatory glutamatergic neurons is a classical mechanism. As is the case throughout the central nervous system, fast synaptic excitation within and between brain regions relevant to epilepsy is mediated predominantly by AMPA receptors. By inhibiting glutamate-mediated excitation, AMPA receptor antagonists markedly reduce or abolish epileptiform activity in in vitro preparations and confer seizure protection in a broad range of animal seizure models. NMDA receptors may also contribute to epileptiform activity, but NMDA receptor blockade is not sufficient to eliminate epileptiform discharges. AMPA receptors move into and out of the synapse in a dynamic fashion in forms of synaptic plasticity, underlying learning and memory. Often, the trigger for these dynamic movements is the activation of NMDA receptors. While NMDA receptor antagonists inhibit these forms of synaptic plasticity, AMPA receptor antagonists do not impair synaptic plasticity and do not inhibit memory formation or retrieval. The demonstrated clinical efficacy of perampanel, a high-potency, orally active non-competitive AMPA receptor antagonist, supports the concept that AMPA receptors are critical to epileptic synchronization and the generation and spread of epileptic discharges in human epilepsy.


Immunization Campaigns and Strategies against Human Papillomavirus in Italy: The Results of a Survey to Regional and Local Health Units Representatives.
Objective. The study aimed to assess the impact of HPV immunization campaigns organizational aspects, the characteristics of immunization program (vaccination targets and type of offer), and communicative strategies adopted by four Italian administrative regions on vaccination coverage observed. Methods. From November 2017 to March 2018, regional and Local Health Units (LHUs) representatives were invited to complete an online survey including 54 questions evaluating vaccination invite systems, access systems to vaccination centres, reminder and recall systems, and adverse events surveillance. An overall descriptive analysis was conducted. Since observed vaccine coverage (VC) obtained in females (2002-2004 birth cohorts) was lower than objectives fixed by the Italian Ministry of Health, variables were assessed using the national VC mean obtained in the 2003 girls birth cohort as outcome. Results. Twenty-six LHUs belonging to 4 Northern and Southern Italian regions participated in the study. Organizational aspects significantly related to VC lower than the national mean were access to vaccine centres without appointment and parents' reservation as appointment planning system. Recall systems for both the first and the second dose, including the appointment in the invitation letter, the availability of regional immunization registry, and education of healthcare workers on universal HPV immunization strategies, instead, were related to higher VC. As regards preadolescent immunization strategies, both VC obtained in girls and boys were far from the Ministerial goals. Only 20% of LHUs introduced multicohort female strategies while all LHUs adopted copayment targeting both men and women. Immunizations strategies targeting subjects at risk were implemented only in half of participating LHUs. Conclusions. VC observed in participating LHUs are largely lower than the national objectives in all anti-HPV vaccine targets. Both organizational and educational strategies have to be implemented to improve the VC goals.


Relationship between the severity of diversion colitis and the composition of colonic bacteria: a prospective study.
Diversion colitis is the inflammation of the excluded segment of the colon in patients undergoing ostomy. It has been suggested that a change in colonic flora may lead to colitis; however, direct evidence for this disease progression is lacking. The aim of this study was to evaluate the relationship between the severity of diversion colitis and the composition of colonic bacteria. We used culture methods and polymerase chain reaction to analyze the colonic microflora of patients who underwent rectal cancer resection with or without diversion ileostomy. In the diversion group, we also evaluated the severity of colonoscopic and pathologic colitis before reversal. This study enrolled 48 patients: 26 in the diversion group and 22 in the control group. Significant differences were observed between the two groups in the levels of Staphylococcus (p=0.038), Enterococcus (p<0.001), Klebsiella (p<0.001), Pseudomonas (p=0.015), Lactobacillus (p=0.038), presence of anaerobes (p=0.019), and Bifidobacterium (p<0.001). A significant correlation between the severity of colitis and bacterial composition was only observed for Bifidobacterium (p=0.005, correlation coefficient=-0.531). The colonic microflora differed significantly between the diversion and control groups. Bifidobacterium was negatively correlated with the severity of diversion colitis.


Surgical Instrument Sets for Special Operations Expeditionary Surgical Teams.
The deployment of surgical assets has been driven by mission demands throughout years of military operations in Iraq and Afghanistan. The transition to the highly expeditious Golden Hour Offset Surgical Transport Team (GHOST- T) now offers highly mobile surgical assets in nontraditional operating rooms; the content of the surgical instrument sets has also transformed to accommodate this change. The 102nd Forward Surgical Team (FST) was attached to Special Operations assigned to southern Afghanistan from June 2015 to March 2016. The focus was to decrease overall size and weight of FST instrument sets without decreasing surgical capability of the GHOST-T. Each instrument set was evaluated and modified to include essential instruments to perform damage control surgery. The overall number of main instrument sets was decreased from eight to four; simplified augmentation sets have been added, which expand the capabilities of any main set. The overall size was decreased by 40% and overall weight decreased by 58%. The cardiothoracic, thoracotomy, and emergency thoracotomy trays were condensed to thoracic set. The orthopedic and amputation sets were replaced with an augmentation set of a prepackaged orthopedic external fixator set). An augmentation set to the major or minor basic sets, specifically for vascular injuries, was created. Through the reorganization of conventional FST surgical instrument sets to maintain damage control capabilities and mobility, the 102nd GHOST-T reduced surgical equipment volume and weight, providing a lesson learned for future surgical teams operating in austere environments.


Psychiatric disorders in candidates for surgery for epilepsy.
To provide a descriptive analysis of the prevalence and pattern of psychiatric morbidity among 300 consecutive epileptic patients refractive to treatment and admitted during a six year period for evaluation of their candidature for surgery. Patients underwent detailed observation of their seizure and standardised psychiatric assessment. Patients were considered to be refractory to treatment if they continued to manifest seizures with an average frequency of at least once every month even with polytherapy using up to three different anti-convulsants for a period of at least two years. Of the 300 patients, 231 had a temporal lobe focus, 43 had a non-temporal lobe focus, and 26 patients had a generalised and multifocal seizure onset. With the DSM-III-R criteria 142 (47.3%) patients emerged as psychiatric cases. A principal axis I diagnosis was made in 88 (29.3%), and an axis II diagnosis (personality disorder) in another 54 (18.0%) patients. The most common axis I diagnosis was anxiety disorders (10.7%). A schizophrenia-like psychosis was seen in 13 (4.3%). Most patients with personality disorders showed dependent and avoidant personality traits. There was a significantly higher psychotic subscore on the present state examination in the temporal than with the non-temporal group of patients. These findings were not significant when compared with patients with a generalised and multifocal seizure disorder. There were no significant findings between the different seizure focus groups on the neurotic subscores. The findings with regard to laterality of seizure focus and the neurotic or psychotic subscores were not significant.


Cutting edge: TIGIT has T cell-intrinsic inhibitory functions.
Costimulatory molecules regulate the functional outcome of T cell activation, and disturbance of the balance between activating and inhibitory signals results in increased susceptibility to infection or the induction of autoimmunity. Similar to the well-characterized CD28/CTLA-4 costimulatory pathway, a newly emerging pathway consisting of CD226 and T cell Ig and ITIM domain (TIGIT) has been associated with susceptibility to multiple autoimmune diseases. In this study, we examined the role of the putative coinhibitory molecule TIGIT and show that loss of TIGIT in mice results in hyperproliferative T cell responses and increased susceptibility to autoimmunity. TIGIT is thought to indirectly inhibit T cell responses by the induction of tolerogenic dendritic cells. By generating an agonistic anti-TIGIT Ab, we demonstrate that TIGIT can inhibit T cell responses directly independent of APCs. Microarray analysis of T cells stimulated with agonistic anti-TIGIT Ab revealed that TIGIT can act directly on T cells by attenuating TCR-driven activation signals.


Drug treatment options for the 2019-new coronavirus (2019-nCoV).
As of January 22, 2020, a total of 571 cases of the 2019-new coronavirus (2019-nCoV) have been reported in 25 provinces (districts and cities) in China. At present, there is no vaccine or antiviral treatment for human and animal coronavirus, so that identifying the drug treatment options as soon as possible is critical for the response to the 2019-nCoV outbreak. Three general methods, which include existing broad-spectrum antiviral drugs using standard assays, screening of a chemical library containing many existing compounds or databases, and the redevelopment of new specific drugs based on the genome and biophysical understanding of individual coronaviruses, are used to discover the potential antiviral treatment of human pathogen coronavirus. Lopinavir /Ritonavir, Nucleoside analogues, Neuraminidase inhibitors, Remdesivir, peptide (EK1), abidol, RNA synthesis inhibitors (such as TDF, 3TC), anti-inflammatory drugs (such as hormones and other molecules), Chinese traditional medicine, such ShuFengJieDu Capsules and Lianhuaqingwen Capsule, could be the drug treatment options for 2019-nCoV. However, the efficacy and safety of these drugs for 2019- nCoV still need to be further confirmed by clinical experiments.


Reproductive biology of an odacine labrid, Odax pullus.
The sexual ontogeny of butterfish Odax pullus was examined in the Hauraki Gulf of New Zealand through histological analysis of gonad material, size and age information and seasonal patterns of sexual maturation. The patterns of gonad development and schedules of male recruitment were established and sexual ontogeny of O. pullus was diagnosed as monandric protogyny, with all males developing from mature females after female-to-male sex reversal. All individuals underwent an immature female phase before maturing as functional females at 228·7-264·8 mm fork length (L(F) ) and at 1·1-1·5 years of age, and there was no evidence of a juvenile bisexual phase. Degenerating mature oogenic elements were found in the gonad lumen of individuals with developing spermatogenic tissue, providing histological evidence for functional protogyny. Sex change was estimated to occur at 359-379 mm L(F) and 2-3 years of age. The diagnosis of monandric protogyny for O. pullus coincided with the pattern of sexual ontogeny seen in the majority of labrids, particularly those of the same clade (tribe Hypsigenyini) and contrasted with that seen in a number of other temperate labrids. This study suggests that the protogynous mode of sexual development in O. pullus is likely to be lineage-specific, i.e. associated with the phylogeny of labrid sexual development, and is not constrained by environmental effects on the evolution of sex change in temperate regions.


Protective Effect of Piper aduncum Capsule on DMBA-induced Breast Cancer in Rats.
The possible protective effect of Piper aduncum capsule on DMBA (dimethylbenz[α]anthracene)-induced breast cancer in rats was assessed by monitoring the tumor and lung metastases incidence and recording hematological and biochemical parameters and frequency of micronuclei. Mammary carcinogenesis was induced in 36 female Holtzman rats by providing a single subcutaneous injection of DMBA. Oral administration of P. aduncum capsule lowered adenocarcinoma and lymph node metastases incidence. Pulmonary metastasis was significantly lowered (P < 0.05). Hematological indicators showed that the triglyceride level was significantly lowered (P < 0.01) and high-density lipoprotein (HDL) level was significantly increased (P < 0.01). Also, P. aduncum capsule significantly lowered the C reactive protein (CRP) level (P < 0.01) and malondialdehyde level (P < 0.05). There was a significant decrease in the frequency of DMBA-induced micronucleated polychromatic erythrocyte (P < 0.01). Considering the antitumorigenic, hypolipidemic, anti-inflammatory, antioxidant, and antigenotoxic properties of P. aduncum capsule, we conclude that it has a protective effect on DMBA-induced breast cancer in rats.


Synthesis of 6,8,9 poly-substituted purine analogue libraries as pro-apoptotic inducers of human leukemic lymphocytes and DAPK-1 inhibitors.
A 18-member library of 6,8,9-poly-substituted purines was prepared from pyrimidines, primary alcohols, and N,N-dimethylamides under basic conditions via a novel one-pot synthetic pathway controlled by amide sizes and the novel analogues were tested against two leukemia cell lines: Jurkat (acute T cell leukemia) and K562 (chronic erythroleukemia) cells. Compounds having a benzoxy group at C6 position of the aromatic ring exhibited antiproliferative activity in Jurkat cells whereas all compounds induced a lower effect on K562 cells. Analysis of cell cycle, Annexin-V staining, and cleavage of initiator caspases assays showed that the active purine analogues induce cell death by apoptosis. Based on these results, a new purine derivative was synthesized, 6-benzyloxy-9-tert-butyl-8-phenyl-9H-purine (6d), which displayed the highest activity of the series against Jurkat cell lines. Finally, (33)P-radiolabeled kinase assays using 96 recombinant human kinases known to be involved in apoptotic events were performed. Just one of the kinases tested, DAPK-1, was inhibited 50% or more by the phenotypic hits at 10 μM, suggesting that the inhibition of this target could be responsible for the induction of cell death by apoptosis. In agreement with the phenotypic results, the most active antiproliferative agent, 6d, displayed also the lowest IC50 value against recombinant DAPK1 (2.5 μM), further supporting the potential role of this protein on the observed functional response. DAPK-1 inhibition led by 6d together with its pro-apoptotic properties against the Jurkat line makes it an interesting candidate to further investigate the role of DAPK1 kinase in triggering apoptosis in cancer cells, a role which is attracting recent interest.


Paternal age effect: Replication in schizophrenia with intriguing dissociation between bipolar with and without psychosis.
Advanced paternal age (APA) is a risk factor for schizophrenia (Sz) and bipolar disorder (BP). Putative mechanisms include heritable genetic factors, de novo mutations, and epigenetic mechanisms. Few studies have explored phenotypic features associated with APA. The Genomic Psychiatry Cohort established a clinically characterized repository of genomic samples from subjects with a Sz-BP diagnosis or unaffected controls, 12,975 with parental age information. We estimated relative risk ratios for Sz, schizoaffective depressed and bipolar types (SA-D, SA-B), and BP with and without history of psychotic features (PF) relative to the control group, comparing each paternal age group to the reference group 20-24 years. All tests were two-sided with adjustment for multiple comparisons. Subjects with fathers age 45+ had significantly higher risk for all diagnoses except for BP w/o PF. APA also bore no significant relation to family psychiatric history. In conclusion, we replicated APA as a risk factor for Sz. To our knowledge, this is the first published report of APA in a BP sample stratified by psychosis history, extending this association only in BP w/PF. This suggests that phenotypic expression of the APA effect in Sz-BP spectrum is psychosis, per se, rather than other aspects of these complex disorders. The lack of a significant relationship between paternal age and familial disease patterns suggests that underlying mechanisms of the paternal age effect may involve a complex interaction of heritable and non-heritable factors. The authors discuss implications and testable hypotheses, starting with a focus on genetic mechanisms and endophenotypic expressions of dopaminergic function. © 2015 Wiley Periodicals, Inc.


Bone targeted therapies for the prevention of skeletal morbidity in men with prostate cancer.
Men with prostate cancer suffer substantially from bone-related complications. Androgen deprivation therapy itself is a cause of loss of bone mineral density and is associated with an increased incidence of osteoporotic fractures. In advanced disease, bone is by far the most common site of metastasis. Complications of bone metastases prominently include pain and the potential for skeletal events such as spinal cord compression and pathologic fractures. Elevated osteoclast activity is an important aspect of the pathophysiology of both treatment-related osteoporosis and skeletal complications due to metastases. The osteoclast is therefore a therapeutic target. Denosumab is a fully human monoclonal antibody to receptor activator of nuclear factor-κ-B ligand that was designed to potently inhibit osteoclast activity and is the central focus of this review. Bisphosphonates, radiopharmaceuticals and systemically-active hormonal agents such as abiraterone acetate and enzalutamide have each been shown to improve skeletal morbidity in specific clinical situations. Denosumab is the only agent that has been shown to prevent osteoporotic fractures in men receiving androgen deprivation therapy and at elevated risk for fracture. It has also demonstrated superiority to the potent bisphosphonate zoledronic acid for the prevention of skeletal-related events in men with castration-resistant prostate cancer metastatic to bone. Efficacy and toxicity data will be discussed.


The Mirage of Long Term Vital Benefice - Risk for the Beginning of Life?
Cord blood is the "life" of the fetus until birth. After delivery, the newborn is "single" and forced to adapt to live using the latest resources provided by the mother. Those who believe that a newborn is just a miniature independent adult are just trusting one of the illusions of secular medicine. Cord blood contains precious cells, stem, red and white blood cells. T cells as a part of white blood cells prevent infections and other illnesses. Cochrane Database 2013 published a study reporting the role of delayed umbilical cord clamping for the benefit of infants. Harvesting of stem cells increases early clamping. So, is prevention better than treatment, speaking about possible pathologies that can occur throughout life? prospective study of newborns in "Alessandrescu-Rusescu" National Institute for Mother and Child Health, Bucharest, Romania, was monitored by their adaptation to extrauterine life, depending on time and technique of clamping. The impact of harvesting stem cells after birth was explored. Of all babies, 8.23% were premature. Maternal pathology (arterial hypertension, diabetes mellitus, infections, thrombophilia) was present in 31.76% of cases. Of the 85 newborns with harvested stem cells, 47% needed assistance in the neonatal intensive care unit (NICU). Birth asphyxia (SA.7) was present in 10.58% of cases. Two protocols with strong recommendations about umbilical cord clamping and harvesting stem cells, respectively, are necessary.


Acute kidney disease and renal recovery: consensus report of the Acute Disease Quality Initiative (ADQI) 16 Workgroup.
Consensus definitions have been reached for both acute kidney injury (AKI) and chronic kidney disease (CKD) and these definitions are now routinely used in research and clinical practice. The KDIGO guideline defines AKI as an abrupt decrease in kidney function occurring over 7 days or less, whereas CKD is defined by the persistence of kidney disease for a period of >90 days. AKI and CKD are increasingly recognized as related entities and in some instances probably represent a continuum of the disease process. For patients in whom pathophysiologic processes are ongoing, the term acute kidney disease (AKD) has been proposed to define the course of disease after AKI; however, definitions of AKD and strategies for the management of patients with AKD are not currently available. In this consensus statement, the Acute Disease Quality Initiative (ADQI) proposes definitions, staging criteria for AKD, and strategies for the management of affected patients. We also make recommendations for areas of future research, which aim to improve understanding of the underlying processes and improve outcomes for patients with AKD.


Matrix metalloproteinase 1 interacts with neuronal integrins and stimulates dephosphorylation of Akt.
Several studies have demonstrated that matrix metalloproteinases (MMPs) are cytotoxic. The responsible mechanisms, however, are not well understood. MMPs may promote cytotoxicity through their ability to disrupt or degrade matrix proteins that support cell survival, and MMPs may also cleave substrates to generate molecules that stimulate cell death. In addition, MMPs may themselves act on cell surface receptors that affect cell survival. Among such receptors is the alpha(2)beta(1) integrin, a complex that has previously been linked to leukocyte death. In the present study we show that human neurons express alpha(2)beta(1) and that pro-MMP-1 interacts with this integrin complex. We also show that stimulation of neuronal cultures with MMP-1 is associated with a rapid reduction in the phosphorylation of Akt, a kinase that can influence caspase activity and cell survival. Moreover, MMP-1-associated dephosphorylation of Akt is inhibited by a blocking antibody to the alpha(2) integrin, but not by batimastat, an inhibitor of MMP-1 enzymatic activity. Such dephosphorylation is also stimulated by a catalytic mutant of pro-MMP-1. Additional studies show that MMP-1 causes neuronal death, which is significantly diminished by both a general caspase inhibitor and anti-alpha(2) but not by batimastat. Together, these results suggest that MMP-1 can stimulate dephosphorylation of Akt and neuronal death through a non-proteolytic mechanism that involves changes in integrin signaling.


Making the most of expert judgment in hazard and risk assessment of chemicals.
Evaluation of the reliability and relevance of toxicity and ecotoxicity studies is an integral step in the assessment of the hazards and risks of chemicals. This evaluation is inherently reliant on expert judgment, which often leads to differences between experts' conclusions regarding how individual studies can contribute to the body of evidence. The conclusions of regulatory assessment, such as establishing safe exposure levels for humans and the environment and calculations of margins of exposure, may have large consequences for which chemicals are permitted on the market and their allowed uses. It is therefore important that such assessments are based on all reliable and relevant scientific data, and that assessment principles and assumptions, such as expert judgment, are transparently applied. It is not possible nor desirable to completely eliminate expert judgment from the evaluation of (eco)toxicity studies. However, it is desirable to introduce measures that increase structure and transparency in the evaluation process so as to provide scientifically robust risk assessments that can be used for regulatory decision making. In this article we present results from workshop exercises with Nordic experts to illustrate how experts' evaluations regarding the reliability and relevance of (eco)toxicity studies for risk assessment may vary and discuss methods intended to promote structure and transparency in the evaluation process.


Trends in the utilization of high-volume hospitals by minority and underinsured surgical patients.
Race- and insurance-based disparities exist in the utilization of high-volume hospitals for complex surgery. Retrospective analysis of the Nationwide Inpatient Sample from 1988 through 2005 was performed to examine hospital volume trends for eight procedures. Ordered logistic regression analyses were performed to determine temporal trends in the utilization of high-volume hospitals by minority and Medicaid-insured patients compared with white patients and those with private insurance or Medicare. Black patients are increasing their utilization of higher-volume hospitals, but not at a rate sufficient to overcome existing disparities relative to the utilization of such hospitals by white patients. Meanwhile, disparities in the utilization of higher-volume hospitals are increasing for Hispanics and patients who are primarily insured through Medicaid. Existing racial and insurance-based disparities in the utilization of high-volume surgical care will persist or become even more pronounced without active intervention from health care policymakers.


Genetic structuring of Patagonian toothfish populations in the Southwest Atlantic Ocean: the effect of the Antarctic Polar Front and deep-water troughs as barriers to genetic exchange.
The environmental and/or life history factors affecting genetic exchange in marine species with potential for high dispersal are of great interest, not only from an evolutionary standpoint but also with regard to effective management. Previous genetic studies have demonstrated substantial differentiation among populations of the Patagonian toothfish around the Southern Ocean, indicating breakdown of gene flow across large distances between inhabited shelf areas. The present study examined genetic structuring through analysis of microsatellite loci and restriction fragment length polymorphism (RFLP) of the mitochondrial ND2 gene and control region of the toothfish population in the SW Atlantic, allowing examination of the relative effects of the Antarctic Polar Front (APF), deep-water troughs and distance between sites. Mitochondrial DNA (mtDNA) data indicated a sharp genetic division between the Patagonian Shelf/North Scotia Ridge and the Shag Rocks/South Georgia samples, whereas microsatellite data showed much less distinct structuring and an intermediate position of the North Scotia Ridge samples. We suggest these data indicate that the APF, as a barrier to larval dispersal, is the major inhibitor of genetic exchange between toothfish populations, with deep-water troughs and distance between sites contributing to genetic differentiation by inhibiting migration of relatively sedentary adults. We also suggest that differences between mtDNA and nuclear DNA population patterns may reflect either genome population size effects or (putative) male-biased dispersal.


Acquisition and functional consequences of social knowledge in macaques.
To manoeuvre in complex societies, it is beneficial to acquire knowledge about the social relationships existing among group mates, so as to better predict their behaviour. Although such knowledge has been firmly established in a variety of animal taxa, how animals acquire such knowledge, as well as its functional significance, remains poorly understood. In order to understand how primates acquire and use their social knowledge, we studied kin-biased redirected aggression in Japanese macaques (Macaca fuscata) relying on a large database of over 15 000 aggressive episodes. Confirming previous research, macaques redirected aggression preferentially to the kin of their aggressor. An analysis that controlled for the rate of affiliation between aggressors and targets of redirection showed that macaques identified the relatives of group mates on the basis of the frequency of their ongoing associations. By contrast, having observed group mates interact with their mother as infants did not increase the monkeys' success in correctly identifying kin relationships among third parties. Inter-individual variation in the successful identification of the kin of aggressors and in redirecting aggression accordingly translated into differences in the amount of aggression received, highlighting a selective advantage for those individuals that were better able to acquire and use social knowledge.


Patterns of co-occurring addictions, posttraumatic stress disorder, and major depressive disorder in detoxification treatment seekers: Implications for improving detoxification treatment outcomes.
Poly-substance use and psychiatric comorbidity are common among individuals receiving substance detoxification services. Posttraumatic stress disorder (PTSD) and major depressive disorder (MDD) are the most common co-occurring psychiatric disorders with substance use disorder (SUD). Current treatment favors a one-size-fits-all approach to treating addiction focusing on one substance or one comorbidity. Research examining patterns of substance use and comorbidities can inform efforts to effectively identify and differentially treat individuals with co-occurring conditions. Using latent class analysis, the current study identified four patterns of PTSD, MDD, and substance use among 375 addiction treatment seekers receiving medically supervised detoxification. The four identified classes were: 1) a PTSD-MDD-Poly SUD class characterized by PTSD and MDD occurring in the context of opioid, cannabis, and tobacco use disorders; 2) an MDD-Poly SUD class characterized by MDD and alcohol, opioid, tobacco, and cannabis use disorders; 3) an alcohol-tobacco class characterized by alcohol and tobacco use disorders; and 4) an opioid-tobacco use disorder class characterized by opioid and tobacco use disorders. The observed classes differed on gender and clinical characteristics including addiction severity, trauma history, and PTSD/MDD symptom severity. The observed classes likely require differing treatment approaches. For example, people in the PTSD-MDD-Poly SUD class would likely benefit from treatment approaches targeting anxiety sensitivity and distress tolerance, while the opioid-tobacco class would benefit from treatments that incorporate motivational interviewing. Appropriate matching of treatment to class could optimize treatment outcomes for polysubstance and comorbid psychiatric treatment seekers. These findings also underscore the importance of well-developed referral networks to optimize outpatient psychotherapy for detoxification treatment-seekers to enhance long-term recovery, particularly those that include transdiagnostic treatment components.


Large-Scale Neuromorphic Spiking Array Processors: A Quest to Mimic the Brain.
Neuromorphic engineering (NE) encompasses a diverse range of approaches to information processing that are inspired by neurobiological systems, and this feature distinguishes neuromorphic systems from conventional computing systems. The brain has evolved over billions of years to solve difficult engineering problems by using efficient, parallel, low-power computation. The goal of NE is to design systems capable of brain-like computation. Numerous large-scale neuromorphic projects have emerged recently. This interdisciplinary field was listed among the top 10 technology breakthroughs of 2014 by the MIT Technology Review and among the top 10 emerging technologies of 2015 by the World Economic Forum. NE has two-way goals: one, a scientific goal to understand the computational properties of biological neural systems by using models implemented in integrated circuits (ICs); second, an engineering goal to exploit the known properties of biological systems to design and implement efficient devices for engineering applications. Building hardware neural emulators can be extremely useful for simulating large-scale neural models to explain how intelligent behavior arises in the brain. The principal advantages of neuromorphic emulators are that they are highly energy efficient, parallel and distributed, and require a small silicon area. Thus, compared to conventional CPUs, these neuromorphic emulators are beneficial in many engineering applications such as for the porting of deep learning algorithms for various recognitions tasks. In this review article, we describe some of the most significant neuromorphic spiking emulators, compare the different architectures and approaches used by them, illustrate their advantages and drawbacks, and highlight the capabilities that each can deliver to neural modelers. This article focuses on the discussion of large-scale emulators and is a continuation of a previous review of various neural and synapse circuits (Indiveri et al., 2011). We also explore applications where these emulators have been used and discuss some of their promising future applications.


Last observation carry-forward and last observation analysis.
Drop-out often occurs in clinical trials with multiple visits and drop-out is often informative in the sense that the population of patients who dropped out is different from the population of patients who completed the study. To handle data with informative drop-out, an intention-to-treat analysis, which evaluates treatment effects over the population of all randomized patients with at least one post-treatment evaluation, is often required by the regulatory agencies. As a popular and simple intention-to-treat analysis, the last observation carry-forward (LOCF) analysis of variance (ANOVA) performs a statistical test for treatment effects by treating the last observation prior to drop-out as the observation from the last visit. Although discussions, examples and limited empirical results about the LOCF analysis can be found, its theoretical property is unclear. We find that the LOCF one-way ANOVA test is actually asymptotically valid (that is, its asymptotic size is equal to the nominal size) in the special but important case where only two treatments are compared and the two treatment groups have the same number of patients, regardless of whether drop-out is informative or not. In other cases, however, the asymptotic size of the LOCF test is different from the nominal size and is often too small when drop-out is informative, which results in a loss in power of detecting treatment effects, a disadvantage to drug companies. We propose an asymptotically valid test for comparing the global means over subpopulations, where each subpopulation contains patients dropping out after a particular visit. Some simulation results are presented to study the finite sample performance of the LOCF test and our proposed test.


An audit of brain tumor patients treated in 5 years at a single institute: Our regional cancer center experience.
Brain tumors constitute a small presentation of all cancers (1.4%) and cancer related deaths (2.5%). Most of the brain tumors are malignant and carry bad prognosis and even if those which are benign still can interfere with brain functions that are required for daily living. We did an audit of brain tumor patients treated in our center and see their prognosis in correlation with different histological types. We analyzed 497 patients treated at our center from June 2007 to June 2012 of different histological types. All the patients underwent complete central nervous system examination and thorough workup including hematological investigations, radiological investigations, and confirmation of histological type by biopsy. These patients were then treated with surgery followed by radiotherapy or upfront curative radiotherapy as per staging and then kept on regular follow-up as per institutional protocol. Majority of patients were astrocytomas (309 patients) followed by a pituitary adenoma (39 patients), oligodendroglioma (33 patients), medulloblastoma (22 patients), arteriovenous malformations (19 patients), craniopharyngioma (16 patients), and recurrent brain tumors (12 patients). There were few cases of central nervous lymphoma, meningioma, melanocytoma, ependymomas, schwannomas, hemangiopericytoma, gliosarcoma, and primitive neuroectodermal tumor. The histological types vary differently regarding presentation, treatment, and prognosis. Astrocytomas are the most common tumors in presentation (309 patients) in which the high-grade astrocytomas carried worse prognosis. Benign tumors such as pituitary adenomas, schwannomas, and craniopharyngiomas had the best prognosis with survival seen up to almost last follow-up.


Symptoms of Major Depressive Disorder Scale: Performance of a Novel Patient-Reported Symptom Measure.
The Symptoms of Major Depressive Disorder Scale (SMDDS) was expressly developed on the basis of qualitative data to directly incorporate patients' voices into evaluation of treatment benefit in major depressive disorder (MDD) clinical trials. To collect quantitative data necessary to refine/optimize the SMDDS and document its psychometric properties. In this multicenter, observational study, participants with clinically diagnosed MDD completed questionnaires in 2 waves. Wave 1 was designed to refine the SMDDS using Rasch measurement evaluations and item reduction analyses. On a subset of wave 1 subjects, 7 to 12 months later, wave 2 further examined item performance and measurement properties. Exploratory factor analyses and assessments of construct validity and reliability (internal consistency and reproducibility) were completed. Using wave 1 data (N = 315; females = 71%, white = 81%, mean age = 44 years), the SMDDS was revised from 36 to 16 items. The Rasch item threshold map indicated that all but 1 item (suicidal ideation) were appropriately ordered. The 207 wave 2 participants were 74% females, 82% white, with a mean age of 45 years. The exploratory factor analyses resulted in a single component (all standardized factor loadings >0.46). Cronbach α was 0.93 and the 7-day test-retest intraclass correlation coefficient (n = 93) was 0.84 (95% confidence interval 0.77-0.89). SMDDS scores discriminated between MDD severity levels. The 16-item SMDDS generated highly reliable scores with substantial evidence of construct validity. On the basis of the evidence of appropriate content validity and sound psychometric performance, the Food and Drug Administration qualified the SMDDS as an outcome measure to support exploratory efficacy endpoints in MDD clinical trials.


Soft tissue masses of the chest wall and axilla: has metastatic melanoma been considered?
Isolated axillary and chest wall soft tissue masses are an uncommon presentation of metastatic cancer. The authors present three patients in whom malignant melanomas metastatic to these sites had been misdiagnosed, leading to inappropriate oncologic treatment planning in all three cases. The presumed diagnoses, even after fine-needle aspiration or trucut biopsies, were soft-tissue sarcoma (n = 2) and undifferentiated breast cancer (n = 1). The combination of taking a thorough history and performing proper immunohistochemical analysis of the biopsy material would have suggested the presence of malignant melanoma in all cases. As the disease appeared locoregionally limited in all patients, radical surgical resection with extended lymphadenectomy was performed without significant dysfunction of the upper extremity. One patient agreed to postoperative immunotherapy with interferon-alpha. Two patients are currently alive 17 and 14 months after operation. One patient was found to have systemic recurrence at 5 months, one experienced two isolated local recurrences in a prior operative site that were amenable to reresection and presently has no evidence of disease 12 months after resection, and one patient remains free of disease at 14 months. Clinical presentation, suggested diagnostic workup, and therapeutic implications are discussed to avoid misdiagnoses in this setting of possible clinical presentations of metastatic melanoma.


Colour vision testing for diabetic retinopathy: a systematic review of diagnostic accuracy and economic evaluation.
To determine the diagnostic performance and cost-effectiveness of colour vision testing (CVT) to identify and monitor the progression of diabetic retinopathy (DR). Major electronic databases including MEDLINE, EMBASE, Cumulative Index to Nursing and Allied Health Literature, and Cochrane Database of Systematic Reviews were searched from inception to September 2008. A systematic review of the evidence was carried out according to standard methods. An online survey of National Screening Programme for Diabetic Retinopathy (NSPDR) clinical leads and programme managers assessed the diagnostic tools used routinely by local centres and their views on future research priorities. A decision tree and Markov model was developed to estimate the incremental costs and effects of adding CVT to the current NSPDR. In total, 25 studies on CVT met the inclusion criteria for the review, including 18 presenting 2 x 2 diagnostic accuracy data. The quality of studies and reporting was generally poor. Automated or computerised CVTs reported variable sensitivities (63-97%) and specificities (71-95%). One study reported good diagnostic accuracy estimates for computerised CVT plus retinal photography for detection of sight-threatening DR, but it included few cases of retinopathy in total. Results for pseudoisochromatic plates, anomaloscopes and colour arrangement tests were largely inadequate for DR screening, with Youden indices (sensitivity + specificity - 100%) close to zero. No studies were located that addressed patient preferences relating to CVT for DR. Retinal photography is universally employed as the primary method for retinal screening by centres responding to the online survey; none used CVT. The review of the economic evaluation literature found no previous studies describing the cost and effects of any type of CVT. Our economic evaluation suggested that adding CVT to the current national screening programme could be cost-effective if it adequately increases sensitivity and is relatively inexpensive. The deterministic base-case analysis indicated that the cost per quality-adjusted life-year gained may be 6364 pounds and 12,432 pounds for type 1 and type 2 diabetes respectively. However, probabilistic sensitivity analysis highlighted the substantial probability that CVT is not diagnostically accurate enough to be either an effective or a cost-effective addition to current screening methods. The results of the economic model should be treated with caution as the model is based on only one small study. There is insufficient evidence to support the use of CVT alone, or in combination with retinal photography, as a method for screening for retinopathy in patients with diabetes. Better quality diagnostic accuracy studies directly comparing the incremental value of CVT in addition to retinal photography are needed before drawing conclusions on cost-effectiveness. The most frequently cited preference for future research was the use of optical coherence tomography for the detection of clinically significant macular oedema.


Evaluation of Factors Affecting Lymph Node Metastasis in Clinical Stage I-II Epithelial Ovarian Cancer.
Systematic lymphadenectomy is useful for accurate staging of early-stage ovarian cancer and has obvious prognostic value. Accurate staging may prevent unnecessary postoperative chemotherapy. The aim of this study was to evaluate the rate of lymph node involvement and factors affecting it in clinically early-stage epithelial ovarian cancer (EOC; stages I, II). The study included 163 patients who underwent surgery at our hospital between January 2004 and April 2017 and who were diagnosed with early-stage EOC based on preoperative and intraoperative examination. Patient data were retrospectively analyzed. The rate of lymph node involvement and factors affecting it were analyzed. Of 163 patients, 21 (12.9%) had lymph node metastasis, whereas 16 (16.3%) of 98 patients who underwent comprehensive lymphadenectomy had lymph node metastasis. According to the univariate results for patients undergoing any type of lymphadenectomy, the rate of positive lymph nodes was significantly higher (37.1%) in those with bilateral ovarian involvement (p < 0.001). The rate was significantly higher in patients with positive intraabdominal fluid cytology (25.9%; p < 0.001), serous histology (20.5%; p = 0.02), and grade 3 disease (33.3%; p < 0.001). In multivariate logistic regression analysis, the rate was significantly higher in patients with bilateral adnexal involvement (p = 0.012). The risk of positive lymph nodes was significantly higher in patients with grade 3 disease (p = 0.016). Comprehensive lymphadenectomy increases the detection rate for metastatic lymph nodes in patients with clinically early-stage EOC. The rate of lymph node involvement is significantly higher in grade 3 tumors, serous cytology, bilateral adnexal involvement, and positive intraabdominal fluid cytology.


The effect of lifelong bilingualism on regional grey and white matter volume.
Lifelong bilingualism is associated with the delayed diagnosis of dementia, suggesting bilingual experience is relevant to brain health in aging. While the effects of bilingualism on cognitive functions across the lifespan are well documented, less is known about the neural substrates underlying differential behaviour. It is clear that bilingualism affects brain regions that mediate language abilities and that these regions are at least partially overlapping with those that exhibit age-related decline. Moreover, the behavioural advantages observed in bilingualism are generally found in executive function performance, suggesting that the frontal lobes may also be sensitive to bilingualism, which exhibit volume reductions with age. The current study investigated structural differences in the brain of lifelong bilingual older adults (n=14, mean age=70.4) compared with older monolinguals (n=14, mean age=70.6). We employed two analytic approaches: 1) we examined global differences in grey and white matter volumes; and, 2) we examined local differences in volume and cortical thickness of specific regions of interest previously implicated in bilingual/monolingual comparisons (temporal pole) or in aging (entorhinal cortex and hippocampus). We expected bilinguals would exhibit greater volume of the frontal lobe and temporal lobe (grey and white matter), given the importance of these regions in executive and language functions, respectively. We further hypothesized that regions in the medial temporal lobe, which demonstrate early changes in aging and exhibit neural pathology in dementia, would be more preserved in the bilingual group. As predicted, bilinguals exhibit greater frontal lobe white matter compared with monolinguals. Moreover, increasing age was related to decreasing temporal pole cortical thickness in the monolingual group, but no such relationship was observed for bilinguals. Finally, Stroop task performance was positively correlated with frontal lobe white matter, emphasizing the importance of preserved white matter in maintaining executive function in aging. These results underscore previous findings implicating an association between bilingualism and preserved frontal and temporal lobe function in aging. This article is part of a Special Issue entitled SI: Memory Å.


Mitochondrial genome sequencing in Mesolithic North East Europe Unearths a new sub-clade within the broadly distributed human haplogroup C1.
The human mitochondrial haplogroup C1 has a broad global distribution but is extremely rare in Europe today. Recent ancient DNA evidence has demonstrated its presence in European Mesolithic individuals. Three individuals from the 7,500 year old Mesolithic site of Yuzhnyy Oleni Ostrov, Western Russia, could be assigned to haplogroup C1 based on mitochondrial hypervariable region I sequences. However, hypervariable region I data alone could not provide enough resolution to establish the phylogenetic relationship of these Mesolithic haplotypes with haplogroup C1 mitochondrial DNA sequences found today in populations of Europe, Asia and the Americas. In order to obtain high-resolution data and shed light on the origin of this European Mesolithic C1 haplotype, we target-enriched and sequenced the complete mitochondrial genome of one Yuzhnyy Oleni Ostrov C1 individual. The updated phylogeny of C1 haplogroups indicated that the Yuzhnyy Oleni Ostrov haplotype represents a new distinct clade, provisionally coined "C1f". We show that all three C1 carriers of Yuzhnyy Oleni Ostrov belong to this clade. No haplotype closely related to the C1f sequence could be found in the large current database of ancient and present-day mitochondrial genomes. Hence, we have discovered past human mitochondrial diversity that has not been observed in modern-day populations so far. The lack of positive matches in modern populations may be explained by under-sampling of rare modern C1 carriers or by demographic processes, population extinction or replacement, that may have impacted on populations of Northeast Europe since prehistoric times.


Disinfectant use as a risk factor for atopic sensitization and symptoms consistent with asthma: an epidemiological study.
Exposure to some nonallergenic compounds has been shown to increase the risk of atopic sensitization and asthmatic symptoms. In order to gain more insight into the largely unknown aetiology of respiratory symptoms in pig farmers, we studied the role of nonallergic exposure. We evaluated associations between chronic respiratory symptoms, specific and total serum immunoglobulin E (IgE) levels, use of disinfectants, and endotoxin exposure levels in a population of 194 Dutch pig farmers. Atopic sensitization (defined as increased production of IgE to common allergens) was found to occur more frequently in farmers who used disinfectants containing quaternary ammonium compounds (QACs) (odds ratio (OR) 7.4; 95% confidence interval (95% CI) 1.3-43.1). ORs for other disinfectants ranged 2.3-4.1 (NS). Atopic sensitization was not found to occur more frequently in farmers with a high endotoxin exposure. The use of disinfectants was only related to respiratory symptoms consistent with asthma in atopics. This is illustrated by the significantly elevated ORs for farmers with IgE to common allergens (house dust mite, grass pollen, birch pollen), and who used disinfectants containing QACs, in the total population and in a subgroup of the total population restricted according to bronchial hyperresponsiveness to histamine (symptomatics with a provocation dose of histamine producing a > or = 10% decrease in forced expiratory volume in one second (PC10) < or = 16 mg.mL-1, compared with asymptomatics with a PC10 > 16 mg.mL-1) (OR 4.4, 95% CI 1.3-14.6; and OR 8.2, 95% CI 1.6-42.6, respectively). Atopy and use of QACs and endotoxin exposure level taken individually were not associated with respiratory symptoms. A combination of atopic sensitization and high endotoxin exposure (> 101 ng.m-3) was strongly associated with respiratory symptoms in the restricted population (OR 6.1; 95% CI 1.0-36.2). Our results suggest that occupational exposure to nonallergenic agents (disinfectants) may induce immunoglobulin E sensitization to common aeroallergens, and that the combination of atopy and exposure to nonallergenic agents (disinfectants and endotoxin) is an important risk factor for development of symptoms consistent with asthma.


Initial management of suspected transient cerebral ischaemia and stroke in primary care: implications of recent research.
Strategies are required to reduce the personal, societal and healthcare burden caused by cerebrovascular disease. Urgent medical intervention after transient ischaemic attack (TIA) can prevent recurrent stroke, and modern healthcare has to respond rapidly to the patient with TIA. The primary care practitioner contributes to stroke prevention by rapidly and accurately diagnosing TIA and arranging urgent specialist assessment. Diagnosis of TIA in primary care is difficult, as transient symptoms are common. Stroke-screening tools are available, but there is no evidence base for diagnostic support tools for TIA in primary care. The ABCD2 scoring system identifies patients after TIA at high early risk, and secondary care assessment within 24 h is reserved for patients with a high predicted risk. General practitioners are advised to give aspirin at the time of diagnosis, although prescribing a full range of vascular risk-reducing therapies may be appropriate. Specialist assessment confirms the diagnosis, usually with cerebral imaging (preferably MRI to detect cerebral injury), and carotid ultrasound will detect patients suitable for endarterectomy. Patients with suspected stroke should be urgently transferred to the nearest stroke centre, for assessment and investigation before potential thrombolysis, which may be effective within a longer timeframe than current practice. Primary care follow-up is essential to ensure adherence to evidence-based therapies. Dual combinations of antiplatelet agents (aspirin and dipyridamole) and antihypertensive agents (ACE inhibitors and thiazides) as well as high-dose statins have proven benefit. For patients in atrial fibrillation, even if very elderly, anticoagulation has a net benefit in preventing stroke.


Project MinE: study design and pilot analyses of a large-scale whole-genome sequencing study in amyotrophic lateral sclerosis.
The most recent genome-wide association study in amyotrophic lateral sclerosis (ALS) demonstrates a disproportionate contribution from low-frequency variants to genetic susceptibility to disease. We have therefore begun Project MinE, an international collaboration that seeks to analyze whole-genome sequence data of at least 15 000 ALS patients and 7500 controls. Here, we report on the design of Project MinE and pilot analyses of successfully sequenced 1169 ALS patients and 608 controls drawn from the Netherlands. As has become characteristic of sequencing studies, we find an abundance of rare genetic variation (minor allele frequency < 0.1%), the vast majority of which is absent in public datasets. Principal component analysis reveals local geographical clustering of these variants within The Netherlands. We use the whole-genome sequence data to explore the implications of poor geographical matching of cases and controls in a sequence-based disease study and to investigate how ancestry-matched, externally sequenced controls can induce false positive associations. Also, we have publicly released genome-wide minor allele counts in cases and controls, as well as results from genic burden tests.