Accurate, sensitive determination of omega-6 and omega-3 polyunsaturated fatty acids in human plasma, urine samples.
Quantitative determination of omega-6 and omega-3 polyunsaturated fatty acids in human plasma and urine with high accuracy and precision provides significant information to monitor the underlying etiology of several diseases. In this regard, liquid chromatography-mass spectrometry is a good choice owing to its great selectivity and sensitivity. Additionally, the hybrid quadrupole-time of flight-mass spectrometer systems provides easy identification of target compounds with superior mass measurements. In this study, an analytical method has been developed for simple, accurate and simultaneous determination of linoleic acid, arachidonic acid, docosahexaenoic acid and eicosapentaenoic acid in a short chromatographic analysis period. The developed method is suitable for the quantitative detection of these four compounds with detection limits ranging between 1.1-3.0 ng ml-1 and its applicability was assessed in human urine and plasma samples. As a result, acceptable accuracy (between 83 and 111%) and good precision (<6%) were obtained for target compounds using matrix matching calibration strategy.


Development of a spatially targeted field sampling technique for the southern cattle tick, Rhipicephalus microplus, by mapping white-tailed deer, Odocoileus virginianus, habitat in South Texas.
The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for deer movement and two vegetation indices: the Normalized Difference Vegetation Index and the Modified Soil Adjusted Vegetation Index to identify forage for deer. These two data sets of favorable white-tailed deer habitat were combined within a geographic information system to identify locations for sampling ticks. Larvae of R. (B.) microplus, were sampled in Zapata County, Texas, by walking transects with attached flannel panels to jeans. Although the data set and sampling period were limited, data analysis demonstrated that sampling of free-living larvae of R. (B.) microplus can be conducted in South Texas, and larvae were most abundant in areas that harbored O. virginianus. Spatial analysis of satellite imagery to classify white-tailed deer/southern cattle tick habitat proved efficacious and may be useful in directing sampling activities in the field.


Origin of initial burst in activity for Trichoderma reesei endo-glucanases hydrolyzing insoluble cellulose.
The kinetics of cellulose hydrolysis have long been described by an initial fast hydrolysis rate, tapering rapidly off, leading to a process that takes days rather than hours to complete. This behavior has been mainly attributed to the action of cellobiohydrolases and often linked to the processive mechanism of this exo-acting group of enzymes. The initial kinetics of endo-glucanases (EGs) is far less investigated, partly due to a limited availability of quantitative assay technologies. We have used isothermal calorimetry to monitor the early time course of the hydrolysis of insoluble cellulose by the three main EGs from Trichoderma reesei (Tr): TrCel7B (formerly EG I), TrCel5A (EG II), and TrCel12A (EG III). These endo-glucanases show a distinctive initial burst with a maximal rate that is about 5-fold higher than the rate after 5 min of hydrolysis. The burst is particularly conspicuous for TrCel7B, which reaches a maximal turnover of about 20 s(-1) at 30 °C and conducts about 1200 catalytic cycles per enzyme molecule in the initial fast phase. For TrCel5A and TrCel12A the extent of the burst is 2-300 cycles per enzyme molecule. The availability of continuous data on EG activity allows an analysis of the mechanisms underlying the initial kinetics, and it is suggested that the slowdown is linked to transient inactivation of enzyme on the cellulose surface. We propose, therefore, that the frequency of structures on the substrate surface that cause transient inactivation determine the extent of the burst phase.


Measurement of antagonist affinity for purine receptors of drugs producing concomitant phosphodiesterase blockade: the use of pharmacologic resultant analysis.
It was observed experimentally that both theophylline and isobutylmethylxanthine (IBMX) produced surmountable antagonism of the agonist effects of 2-chloroadenosine on purine receptors in rat vas deferens and guinea pig atria. In the case of IBMX, there was a statistically significant difference between the Schild regressions in the two tissues, ostensibly indicating a possible purine receptor heterogeneity with respect to the binding of this antagonist. However, the analysis was complicated by the fact that both theophylline and IBMX produced positive inotropic responses in guinea pig atria (presumably by inhibition of cardiac phosphodiesterase), making the calculation of dose ratios subjective and ambiguous. To determine whether the phosphodiesterase blocking property of theophylline and IBMX was interfering with the observation of purine receptor antagonism in guinea pig atria, a new technique described as pharmacologic resultant analysis was utilized to measure the purine receptor blocking properties of theophylline and IBMX in guinea pig atria in the presence of phosphodiesterase blockade. Based on the principle of additive dose ratios for two antagonists competing for one receptor, pharmacologic resultant analysis measures the effects of a test antagonist (in this case theophylline or IBMX) on the competitive blockade of a reference antagonist; for these studies, the reference antagonist was 8-sulfophenyltheophylline. Under these circumstances, the direct effects of the test antagonist (positive inotropy) are expressed throughout the experiment and cancel with the normal null methods used for measurement of competitive antagonism. Using this technique, it was found that the potencies of both theophylline and IBMX on the purine receptors in rat vas deferens and guinea pig atria were identical. The use of this method in the delineation of multiple drug activities is discussed.


Size-Selective Phagocytic Clearance of Fibrillar α-Synuclein through Conformational Activation of Complement Receptor 4.
Aggregation of α-synuclein (αSN) is an important histological feature of Parkinson disease. Recent studies showed that the release of misfolded αSN from human and rodent neurons is relevant to the progression and spread of αSN pathology. Little is known, however, about the mechanisms responsible for clearance of extracellular αSN. This study found that human complement receptor (CR) 4 selectively bound fibrillar αSN, but not monomeric species. αSN is an abundant protein in the CNS, which potentially could overwhelm clearance of cytotoxic αSN species. The selectivity of CR4 toward binding fibrillar αSN consequently adds an important αSN receptor function for maintenance of brain homeostasis. Based on the recently solved structures of αSN fibrils and the known ligand preference of CR4, we hypothesize that the parallel monomer stacking in fibrillar αSN creates a known danger-associated molecular pattern of stretches of anionic side chains strongly bound by CR4. Conformational change in the receptor regulated tightly clearance of fibrillar αSN by human monocytes. The induced change coupled concomitantly with phagolysosome formation. Data mining of the brain transcriptome in Parkinson disease patients supported CR4 as an active αSN clearance mechanism in this disease. Our results associate an important part of the innate immune system, namely complement receptors, with the central molecular mechanisms of CNS protein aggregation in neurodegenerative disorders.


Hypothalamic lipids and the regulation of energy homeostasis.
The hypothalamus is a specialised area in the brain that integrates the control of energy homeostasis, regulating both food intake and energy expenditure. The classical theory for hypothalamic feeding control is mainly based on the relationship between peripheral signals and neurotransmitters/neuromodulators in the central nervous system. Thus, hypothalamic neurons respond to peripheral signals, such as hormones and nutrients, by modifying the synthesis of neuropeptides. Despite the well-established role of these hypothalamic networks, increasing evidence indicates that the modulation of lipid metabolism in the hypothalamus plays a critical role in feeding control. In fact, the pharmacologic and genetic targeting of key enzymes from these pathways, such as AMP-activated protein kinase, acetyl-CoA carboxylase, carnitine palmitoyltransferase 1, fatty acid synthase, and malonyl-CoA decarboxylase, has a profound effect on food intake and body weight. Here, we review what is currently known about the relationship between hypothalamic lipid metabolism and whole body energy homeostasis. Defining these novel mechanisms may offer new therapeutic targets for the treatment of obesity and its associated pathologies.


Predictors of quality of life in patients with diabetes mellitus in two tertiary health institutions in Ghana and Nigeria.
Patients with chronic diseases such as Type 2 diabetes mellitus (DM) usually have a relatively poor quality of life (QoL), because the cost of care (living expenses and health) or diet restrictions are heavily felt by these patients, and this is of a public health concern. However, limited data on DM QoL exist in Ghana and Nigeria. This makes it imperative for data to be collated in that regard. We adopted the Strengthening The reporting of observational studies in epidemiology (STROBE) consensus checklist to survey the patients with DM seen at the diabetic clinic at the Department of Medicine of the Korle-Bu Teaching Hospital and University College Hospital, Ibadan, Nigeria. Patients with Type 2 DM aged 40 years and older were recruited by using systematic random sampling method. The World Health Organization Quality of Life-BREF, diabetes empowerment scale, and DM knowledge scale were used to assess QoL, patient empowerment, and knowledge of DM, respectively. The predictors of QoL were determined using multiple linear regression analyses. A total of 198 patients in Ghana and 203 patients in Nigeria completed the survey, with female-to-male ratio being 3:1 and 2:1, respectively. The overall QoL in both countries was relatively low: 56.19 ± 8.23 in Ghana and 64.34 ± 7.34 in Nigeria. In Ghana, significant correlates of higher scores on the QoL scale were medication adherence (P = 0.02) and employment status (P = 0.02). Among patients in Nigeria, employment status (P = 0.02) and DM empowerment (0.03) were significant predictors of QoL in patients with DM. Our study revealed an association between a number of psychosocial factors and QoL among patients with DM in Ghana and Nigeria.


Adsorption and Interfacial Layer Structure of Unmodified Nanocrystalline Cellulose at Air/Water Interfaces.
Nanocrystalline cellulose (NCC) is a promising biological nanoparticle for the stabilization of fluid interfaces. However, the adsorption and interfacial layer structure of NCC are poorly understood as it is currently unknown how to form NCC interfacial layers. Herein, we present parameters for the adsorption of unmodified NCC at the air-water (A/W) interface. Initial NCC adsorption is limited by diffusion, followed by monolayer saturation and decrease in surface tension at the time scale of hours. These results confirm the current hypothesis of a Pickering stabilization. NCC interfacial performance can be modulated by salt-induced charge screening, enhancing adsorption kinetics, surface load, and interfacial viscoelasticity. Adsorbed NCC layers were visualized by atomic force microscopy at planar Langmuir films and curved air bubbles, whereat NCC coverage was higher at curved interfaces. Structural analysis by neutron reflectometry revealed that NCC forms a discontinuous monolayer with crystallites oriented in the interfacial plane at a contact angle < 90°, favoring NCC desorption upon area compression. This provides the fundamental framework on the formation and structure of NCC layers at the A/W interface, paving the way for exploiting NCC interfacial stabilization for tailored colloidal materials.


First Report of Chilli ringspot virus on Chili Pepper in China.
Viral diseases have been a major limiting factor in the production of chili pepper (Capsicum chinense Jacp. cv. Yellow Lantern) in Hainan Province, China. In a 2009 disease survey, we found heavily infected fields of chili pepper exhibiting typical viral disease symptoms in three counties in Hainan. Total RNA was extracted from leaves of 14 randomly sampled plants and used as templates for reverse transcription (RT)-PCR using a pair of primers (forward 5' CGTAGACAACACACTCATGGT 3', reverse 5' GTTTTCCCAGTCACGAC(T)16 3') that were originally designed to detect Chilli veinal mottle virus (ChiVMV) (2). PCR fragments of 1.6 kb were amplified from the diseased but not the symptom-free samples and were subsequently sequenced. While most samples were found to be infected with ChiVMV, sequences of PCR fragments from three samples showed identities greater than 90% to two sequences of Chilli ringspot virus (ChiRSV), a member of Potyviridae, available in GenBank (Accession Nos. DQ925439 and DQ925438) (1), but less than 60% to that of the Hainan isolate of ChiVMV (Accession No. GQ981316), a distinct potyvirus (2). Furthermore, two of the samples contained one ChiRSV genotype while the third sample contained a different ChiRSV genotype, with a sequence identity of 91.7% between the two. Using these sequences, we subsequently designed a pair of primers (forward 5' TGGGATAGAGCATCTGAGC 3' and reverse 5' GAGTCATTTAGGTCATAATCAGTTT 3') for specific amplification of ChiRSV but not ChiVMV by RT-PCR. A ChiRSV-specific 0.6-kb DNA fragment was amplified from 8 of the 14 samples. Sequencing of these amplicons confirmed the presence of ChiRSV in these samples. Flexuous, rod-shaped virus particles, typical of the potyviruses, were observed by electron microscopy in the virion preparations purified from chili plants infected only with ChiRSV. Virion protein was purified and subjected to analysis by the MALDI-TOF/TOF tandem mass spectrometer (BGI, Shenzhen, China), yielding 35 peptide fragments that share the highest sequence homology to the coat protein of Vietnamese ChiRSV C8 and C9 isolates (GenBank Accession Nos. ABL09413 and ABL09414) (1) with protein sequence coverage of 61.8% and 52.9%, respectively. On the basis of these data, we concluded that the chili plants in Hainan were infected with ChiRSV. This virus was first reported in Vietnam in 2008 (1) but has not been found elsewhere. Leaves of the chili plants infected only with the Hainan isolate of ChiRSV displayed green banding along the main and major lateral veins, light interveinal chlorosis, and crinkling, similar to the symptoms of ChiVMV previously reported in Hainan (2). However, ChiVMV-infected leaves were notably more distorted than those infected by ChiRSV. We did not observe the characteristic ringspots described on ChiRSV-infected chili peppers in Vietnam (1). The difference in the symptoms may be accounted for by different ChiRSV genotypes, chili varieties, or a combination of both. The finding of two distinct viruses causing similar symptoms will help to improve identification and management of viral diseases on chili peppers. References: (1) C. Ha et al. Arch. Virol. 153:45, 2008. (2) J. Wang et al. Plant Dis. 90:377, 2006.


Stroop facilitation in tinnitus patients: an experiment conducted via the world wide web.
Cognitive mechanisms have been proposed to play an important role in tinnitus. In the present study, tinnitus participants were administered an emotional Stroop test via the Internet, incorporating words related to tinnitus concerns. In line with previous research using this Web-based version of the emotional Stroop test, faster color naming was demonstrated for concern-relevant words relative to neutral words. The present results thus provided support for a role for cognitive factors that are important for the understanding of tinnitus. However, future research is warranted in order to clarify the precise mechanisms involved in tinnitusrelated Stroop effects.


Genetic differentiation in Tetranychus urticae (Acari: Tetranychidae): polymorphism, host races or sibling species?
Based on allozyme electrophoresis at the Pgm locus and nuclear ribosomal DNA (ITS2) sequences, we studied the genetic variation of the two-spotted spider mite Tetranychus urticae Koch collected on rose bay, Nerium oleander L. (Apocynaceae), from several localities around the Mediterranean basin. In addition, we compared these results with those of Navajas et al. (1998) and Tsagkarakou (1997) who collected from several other host plants from the Mediterranean. In the western part of this area (Spain, France, Tunisia), we found the individuals collected from rose bay to be clearly genetically differentiated from other samples. No evidence of such host-associated differentiation was detected in the eastern Mediterranean (Italy and Greece). The genetic differentiation of mites collected on rose bay was investigated further by studying the reproductive incompatibilities between populations in Greece and in France and a laboratory strain reared on bean, Phaseolus vulgaris, in France. Reciprocal crosses performed between these strains revealed variable levels of incompatibility, spanning from partial to complete reproductive isolation. In all cases incompatibility was asymmetric. We designed a test based on double-mating to establish the fertilization status of females in fully incompatible crosses. These crosses showed that the females had been inseminated, which suggests that the barrier to reproduction is not of a prezygotic behavioral nature. The data raises the question of the relative role of ecological factors (host plant) and geographical distance, in the ongoing differentiation process potentially leading to speciation.


Leukemic transformation and second cancers in 3649 patients with high-risk essential thrombocythemia in the EXELS study.
EXELS, a post-marketing observational study, is the largest prospective study of high-risk essential thrombocythemia (ET) patients, with an observation time of 5 years. EXELS found higher event rates of acute leukemia transformation in patients treated with hydroxycarbamide (HC). In the current analysis, we report age-adjusted rates of malignant transformation from 3460 EXELS patients exposed to HC, anagrelide (ANA), or both. At registration, 481 patients had ANA treatment without HC exposure, 2305 had HC without ANA exposure, and 674 had been exposed to both. Standard incidence ratios (SIRs) were calculated using data from the Cancer Incidence in Five Continents database to account for differences in age-, gender-, and country-specific background rates. SIRs for acute myelogenous leukemia (AML) were high in ET patients. SIRs for AML were high in HC-treated patients, but AML was rare in ANA-treated patients; no cases of AML were found in patients only treated with ANA. No statistically significant difference was seen between SIRs for ANA and HC treatment for AML or skin cancer. SIRs for other cancers were similar in the HC and ANA groups and close to 1, indicating little difference in risk. Although statistically inconclusive, this study strengthens concerns regarding possible leukemogenic risk with HC treatment. (NCT00202644).


REMOD: A Tool for Analyzing and Remodeling the Dendritic Architecture of Neural Cells.
Dendritic morphology is a key determinant of how individual neurons acquire a unique signal processing profile. The highly branched dendritic structure that originates from the cell body, explores the surrounding 3D space in a fractal-like manner, until it reaches a certain amount of complexity. Its shape undergoes significant alterations under various physiological or neuropathological conditions. Yet, despite the profound effect that these alterations can have on neuronal function, the causal relationship between the two remains largely elusive. The lack of a systematic approach for remodeling neural cells and their dendritic trees is a key limitation that contributes to this problem. Such causal relationships can be inferred via the use of large-scale neuronal models whereby the anatomical plasticity of neurons is accounted for, in order to enhance their biological relevance and hence their predictive performance. To facilitate this effort, we developed a computational tool named REMOD that allows the structural remodeling of any type of virtual neuron. REMOD is written in Python and can be accessed through a dedicated web interface that guides the user through various options to manipulate selected neuronal morphologies. REMOD can also be used to extract meaningful morphology statistics for one or multiple reconstructions, including features such as sholl analysis, total dendritic length and area, path length to the soma, centrifugal branch order, diameter tapering and more. As such, the tool can be used both for the analysis and/or the remodeling of neuronal morphologies of any type.


An element downstream of the cap site is required for transcription of the gene encoding mouse ribosomal protein L32.
To identify the elements that regulate transcription of the mouse gene encoding ribosomal protein L32 (rpL32), we transfected monkey kidney (COS or CV-1) cells with mutants bearing progressive 5' deletions or an internal deletion in exon I and measured their transient expression by S1 nuclease protection analysis. When the mutant genes were tested in the vector pi SVHSplac, which contains a short segment of the oriregion of simian virus 40, maximum expression was observed with as little as 36 base pairs of 5' flanking sequence, and the mutant bearing the exon I deletion was expressed very efficiently. However, when the genes were tested in a simple prokaryotic (pUC) vector, the expression was increased 3- to 4-fold by sequences between -36 and -159, and the exon I segment was absolutely required for expression. Gel mobility-shift and methylation interference analyses revealed that a nuclear factor specifically binds to a GGCTGCCATC sequence within this exon I segment. These results, taken together with other recent findings, indicate that the elements involved in transcriptional regulation of the rpL32 gene are distributed over a 200-base-pair region that spans the cap site. The contributions of some of these elements are apparently masked in the presence of simian virus 40 ori-region elements.


Scaffolding and working together: a qualitative exploration of strategies for everyday life with dementia.
living with dementia has been described as a process of continual change and adjustment, with people with dementia and their families adopting informal strategies to help manage everyday life. As dementia progresses, families increasingly rely on help from the wider community and formal support. this article reports on a secondary analysis of qualitative data from focus groups and individual interviews with people with dementia and their carers in the North of England. In total, 65 people with dementia and 82 carers took part in the research: 26 in interviews and 121 in focus groups. Focus group and interview audio recordings were transcribed verbatim. A qualitative, inductive, thematic approach was taken for data analysis. the article applies the metaphor of scaffolding to deepen understanding of the strategies used by families. Processes of scaffolding were evident across the data where families, communities, professionals and services worked together to support everyday life for people with dementia. Within this broad theme of scaffolding were three sub-themes characterising the experiences of families living with dementia: doing things together; evolving strategies; and fragility and fear of the future. families with dementia are resourceful but do need increasing support (scaffolding) to continue to live as well as possible as dementia progresses. More integrated, proactive work is required from services that recognises existing scaffolds and provides appropriate support before informal strategies become unsustainable; thus enabling people with dementia to live well for longer.


Predictive models for population performance on real biological fitness landscapes.
Directed evolution, in addition to its principal application of obtaining novel biomolecules, offers significant potential as a vehicle for obtaining useful information about the topologies of biomolecular fitness landscapes. In this article, we make use of a special type of model of fitness landscapes-based on finite state machines-which can be inferred from directed evolution experiments. Importantly, the model is constructed only from the fitness data and phylogeny, not sequence or structural information, which is often absent. The model, called a landscape state machine (LSM), has already been used successfully in the evolutionary computation literature to model the landscapes of artificial optimization problems. Here, we use the method for the first time to simulate a biological fitness landscape based on experimental evaluation. We demonstrate in this study that LSMs are capable not only of representing the structure of model fitness landscapes such as NK-landscapes, but also the fitness landscape of real DNA oligomers binding to a protein (allophycocyanin), data we derived from experimental evaluations on microarrays. The LSMs prove adept at modelling the progress of evolution as a function of various controlling parameters, as validated by evaluations on the real landscapes. Specifically, the ability of the model to 'predict' optimal mutation rates and other parameters of the evolution is demonstrated. A modification to the standard LSM also proves accurate at predicting the effects of recombination on the evolution.


Predictors of mortality in patients with lacunar stroke in the secondary prevention of small subcortical strokes trial.
The Secondary Prevention of Small Subcortical Stroke trial (SPS3) recruited participants meeting clinical and radiological criteria for symptomatic lacunes. Individuals randomized to dual antiplatelet therapy with clopidogrel and aspirin had an unanticipated increase in all-cause mortality compared with those assigned to aspirin. We investigated the factors associated with mortality in this well-characterized population. We identified independent predictors of mortality among baseline demographic and clinical factors by Cox regression analysis in participants of the SPS3 trial. Separately, we examined the effect on mortality of nonfatal bleeding during the trial. During a mean follow-up of 3.6 years, the mortality rate was 1.78% per year for the 3020 participants (mean age, 63 years). Significant independent predictors of mortality at study entry were age, diabetes mellitus, history of hypertension, systolic blood pressure (hazard ratio [HR], 1.3 per 20 mm Hg increase), serum hemoglobin<13 g/dL (HR, 1.6), renal function (HR, 1.3 per estimated glomerular filtration rate decrease of 20 mL/min), and body mass index (HR, 1.8 per 10 kg/m2 decrease). Participants with ischemic heart disease (P=0.01 for interaction) and normotensive/prehypertensive participants (P=0.03 for interaction) were at increased risk if assigned to dual antiplatelet therapy. Nonfatal major hemorrhage increased mortality in both treatment arms (HR, 4.5; 95% confidence interval, 3.1-6.6; P<0.001). Unexpected interactions between assigned antiplatelet therapy and each of ischemic heart disease and normal/prehypertensive status accounted for increased mortality among patients with recent lacunar stroke given dual antiplatelet therapy. Despite extensive exploratory analyses, the mechanisms underlying these interactions are uncertain. http://www.SPS3ClinicalTrials.gov. Unique identifier: NCT00059306.


Engineering Strategies to Decode and Enhance the Genomes of Coral Symbionts.
Elevated sea surface temperatures from a severe and prolonged El Niño event (2014-2016) fueled by climate change have resulted in mass coral bleaching (loss of dinoflagellate photosymbionts, Symbiodinium spp., from coral tissues) and subsequent coral mortality, devastating reefs worldwide. Genetic variation within and between Symbiodinium species strongly influences the bleaching tolerance of corals, thus recent papers have called for genetic engineering of Symbiodinium to elucidate the genetic basis of bleaching-relevant Symbiodinium traits. However, while Symbiodinium has been intensively studied for over 50 years, genetic transformation of Symbiodinium has seen little success likely due to the large evolutionary divergence between Symbiodinium and other model eukaryotes rendering standard transformation systems incompatible. Here, we integrate the growing wealth of Symbiodinium next-generation sequencing data to design tailored genetic engineering strategies. Specifically, we develop a testable expression construct model that incorporates endogenous Symbiodinium promoters, terminators, and genes of interest, as well as an internal ribosomal entry site from a Symbiodinium virus. Furthermore, we assess the potential for CRISPR/Cas9 genome editing through new analyses of the three currently available Symbiodinium genomes. Finally, we discuss how genetic engineering could be applied to enhance the stress tolerance of Symbiodinium, and in turn, coral reefs.


Limited effect of CpG ODN in preventing type 1 diabetes in NOD mice.
Type 1 diabetes is considered as Th1 cell mediated autoimmune disease and the suppression of Th1 cells or the activation of Th2 cells has been regarded as a plausible immunologic intervention for the prevention of type 1 diabetogenesis in a rodent model. CpG ODN is an immunostimulatory sequence primarily present in bacterial DNA, viral DNA and BCG. CpG ODN is conventionally classified as a Th1 cell activator, which has been clinically applied to cancer, allergy and infectious disease. Recently, there was a promising report of that CpG ODN administration suppressed the development of type 1 diabetes in NOD mice by inducing Th2 cell mediated cytokine. However, the antidiabetogenic effect of CpG ODN on NOD mice is controversial. Thus, two studies were serially undertaken with various kinds of CpG motif to find a more optimal sequence and administration method. In the first study, CpG ODN was vaccinated four times and pancreatic inflammation and the quantity of serum insulin subsequently evaluated. In the second study, the amounts of IFN gamma and IL-4 in sera were measured as representative cytokines of Th1 and Th2 cells, respectively. As a result, vaccination or continuous injection of CpG ODN failed to show a preventive effect on type 1 diabetogenesis in NOD mice. Structural differences of CpG ODN also had no affect on the result. CpG ODN also consistently showed affect on the pancreatic pathology. The productions of IFN gamma and IL-4 were detected only in the K and D type CpG ODN administration groups. Comparison of the two cytokines leads to the conclusion that CpG ODN generated a Th1-weighted response in both study groups. It was assumed that CpG ODN failed to produce Th2-weighted cytokine milieu, which can overcome the genetically determined phenotype of NOD mice. Given these results, it was concluded that the immunotherapeutic application of CpG ODN on Type 1 diabetes had clear limitations.


Student-perceived factors for an enhanced advanced education program in prosthodontics recall system.
A qualitative study of Advanced Education Programs in Prosthodontics (AEPPs) students was conducted to identify best practices to effectively promote ongoing health and student learning within the context of a patient-centered recall system. Ten students from seven AEPPs nationwide were invited to participate in a focus group on recall systems within AEPPs. The discussion first identified whether an active recall program existed and then delved into benefits for patients and students, positive and negative features of existing recall systems, and factors that can be improved upon for an enhanced recall system. Participants advocated the highest standard of patient care, including regular ongoing care once restorative therapy is complete. Discussion indicates that not only does regular patient recall lead to health promotion, disease prevention, and monitoring of existing prostheses for the patient, but also provides for an enhanced learning experience for the students. Recognizing this, several students from AEPPs lacking an official recall system have established a "makeshift" system, encompassing a treatment completion letter, final intraoral photographs, patient education, and regular prosthetic evaluations, for their existing patients. Prosthodontic program students perceived their program's recall effectiveness could be improved. Due to the numerous potential benefits of an active recall system for both patients and students, some perceived factors to be improved upon include treatment completion protocol, patient education, and establishment of a patient-centered recall system managed by a team of hygienists, receptionists, attending faculty, and residents.


Metabolic Regulation of the Epigenome Drives Lethal Infantile Ependymoma.
Posterior fossa A (PFA) ependymomas are lethal malignancies of the hindbrain in infants and toddlers. Lacking highly recurrent somatic mutations, PFA ependymomas are proposed to be epigenetically driven tumors for which model systems are lacking. Here we demonstrate that PFA ependymomas are maintained under hypoxia, associated with restricted availability of specific metabolites to diminish histone methylation, and increase histone demethylation and acetylation at histone 3 lysine 27 (H3K27). PFA ependymomas initiate from a cell lineage in the first trimester of human development that resides in restricted oxygen. Unlike other ependymomas, transient exposure of PFA cells to ambient oxygen induces irreversible cellular toxicity. PFA tumors exhibit a low basal level of H3K27me3, and, paradoxically, inhibition of H3K27 methylation specifically disrupts PFA tumor growth. Targeting metabolism and/or the epigenome presents a unique opportunity for rational therapy for infants with PFA ependymoma.


Nutritional Care of Patients Admitted to Hospital for Alcohol Withdrawal: A 5-Year Retrospective Audit.
The aim of this study was to describe the characteristics and the nutritional approaches implemented with patients undergoing alcohol withdrawal. A retrospective analysis of medical records for patients admitted to a tertiary hospital for alcohol withdrawal was completed over a 5-year period 2013-2017. Data on nutrition-related assessment and management were extracted and descriptively analysed. A total of 109 medical records were included (M = 73, F = 36), with the mean age of patients 47.3 years (SD ± 11.2, range 22-70). The average length of stay was 3.7 days (SD ± 3.9, range 0.70-27.8). Approaches towards nutritional care emerged from micronutrient assessment and supplementation and/or dietetic consultation. Nutrition-related biochemistry data was available for most patients, notably serum levels of sodium, urea and creatinine (102 patients; 93.5%) and magnesium and phosphate (66 patients, 60.5%). There was evidence of some electrolyte abnormalities on admission to hospital. Eight patients had serum micronutrient status assessed; no patients had serum thiamine levels assessed. Parenteral thiamine was provided to 96 patients (88.0%) for 1.9 days (SD ± 1.1, range 1.0-6.0) with a mean dose of 2458.7 mg (SD ± 1347.6, range 300-6700 mg). Multivitamin supplementation was provided to 24 patients (22.0%). Only 23 patients (21.2%) were seen by a dietician of whom 16 underwent a comprehensive nutritional assessment and 3 were screened using the malnutrition screening tool. Inconsistent nutritional assessment and management practices were identified across a diverse population group, whilst nutritional professionals were underutilized. Future research should benchmark current guidelines and multidisciplinary approaches considering the role of nutritional specialists in the team.


The National Nursing Assistant Survey: improving the evidence base for policy initiatives to strengthen the certified nursing assistant workforce.
This study introduces the first National Nursing Assistant Survey (NNAS), a major advance in the data available about certified nursing assistants (CNAs) and a rich resource for evidence-based policy, practice, and applied research initiatives. We highlight potential uses of this new survey using select population estimates as examples of how the NNAS can be used to inform new policy directions. The NNAS is a nationally representative survey of 3,017 CNAs working in nursing homes, who were interviewed by phone in 2004-2005. Key survey components are recruitment; education; training and licensure; job history; family life; management and supervision; client relations; organizational commitment and job satisfaction; workplace environment; work-related injuries; and demographics. One in three CNAs received some kind of means-tested public assistance. More than half of CNAs incurred at least 1 work-related injury within the past year and almost one quarter were unable to work for at least 1 day due to the injury. Forty-two percent of uninsured CNAs cite not participating in their employer-sponsored insurance plan because they could not afford the plan. Years of experience do not translate into higher wages; CNAs with 10 or more years of experience averaged just $2/hr more than aides who started working in the field less than 1 year ago. This survey can be used to understand CNA workforce issues and challenges and to plan for sustainable solutions to stabilize this workforce. The NNAS can be linked to other existing data sets to examine more comprehensive and complex relationships among CNA, facility, resident, and community characteristics, thereby expanding its usefulness.


Transcriptome analysis reveals microvascular endothelial cell-dependent pericyte differentiation.
Microvascular homeostasis is strictly regulated, requiring close interaction between endothelial cells and pericytes. Here, we aimed to improve our understanding of how microvascular crosstalk affects pericytes. Human-derived pericytes, cultured in absence, or presence of human endothelial cells, were studied by RNA sequencing. Compared with mono-cultured pericytes, a total of 6704 genes were differentially expressed in co-cultured pericytes. Direct endothelial contact induced transcriptome profiles associated with pericyte maturation, suppression of extracellular matrix production, proliferation, and morphological adaptation. In vitro studies confirmed enhanced pericyte proliferation mediated by endothelial-derived PDGFB and pericyte-derived HB-EGF and FGF2. Endothelial-induced PLXNA2 and ACTR3 upregulation also triggered pericyte morphological adaptation. Pathway analysis predicted a key role for TGFβ signaling in endothelial-induced pericyte differentiation, whereas the effect of signaling via gap- and adherens junctions was limited. We demonstrate that endothelial cells have a major impact on the transcriptional profile of pericytes, regulating endothelial-induced maturation, proliferation, and suppression of ECM production.


Primary blast lung injury prevalence and fatal injuries from explosions: insights from postmortem computed tomographic analysis of 121 improvised explosive device fatalities.
Primary blast lung injury (PBLI) is an acknowledged cause of death in explosive blast casualties. In contrast to vehicle occupants following an in-vehicle explosion, the injury profile, including PBLI incidence, for mounted personnel following an external explosion has yet to be as well defined. This retrospective study identified 146 cases of UK military personnel killed by improvised explosive devices (IEDs) between November 2007 and July 2010. With the permission of Her Majesty's Coroners, relevant postmortem computed tomography imaging was analyzed. PBLI was diagnosed by postmortem computed tomography. Injury, demographic, and relevant incident data were collected via the UK Joint Theatre Trauma Registry. Autopsy results were not available for 1 of 146 cases. Of the remaining 145 IED fatalities, 24 had catastrophic injuries (disruptions), making further study impossible, leaving 121 cases; 79 were dismounted (DM), and 42 were mounted (M). PBLI was noted in 58 cases, 33 (79%) of 42 M fatalities and 25 (32%) of 79 DM fatalities (p < 0.0001). Rates of associated thoracic trauma were also significantly greater in the M group (p < 0.006 for all). Fatal head (53% vs. 23%) and thoracic trauma (23% vs. 8%) were both more common in the M group, while fatal lower extremity trauma (7% vs. 48%) was more commonly seen in DM casualties (p < 0.0001 for all). Following IED strikes, mounted fatalities are primarily caused by head and chest injuries. Lower extremity trauma is the leading cause of death in dismounted fatalities. Mounted fatalities have a high incidence of PBLI, suggesting significant exposure to primary blast. This has not been reported previously. Further work is required to determine the incidence and clinical significance of this severe lung injury in explosive blast survivors. In addition, specific characteristics of the vehicles should be considered.


Enhancing therapeutic vaccination by blocking PD-1-mediated inhibitory signals during chronic infection.
Therapeutic vaccination is a potentially promising strategy to enhance T cell immunity and viral control in chronically infected individuals. However, therapeutic vaccination approaches have fallen short of expectations, and effective boosting of antiviral T cell responses has not always been observed. One of the principal reasons for the limited success of therapeutic vaccination is that virus-specific T cells become functionally exhausted during chronic infections. We now provide a novel strategy for enhancing the efficacy of therapeutic vaccines. In this study, we show that blocking programmed death (PD)-1/PD-L1 inhibitory signals on exhausted CD8(+) T cells, in combination with therapeutic vaccination, synergistically enhances functional CD8(+) T cell responses and improves viral control in mice chronically infected with lymphocytic choriomeningitis virus. This combinatorial therapeutic vaccination was effective even in the absence of CD4(+) T cell help. Thus, our study defines a potent new approach to augment the efficacy of therapeutic vaccination by blocking negative signals. Such an approach may have broad applications in developing treatment strategies for chronic infections in general, and perhaps also for tumors.


Involvement in rehabilitative care and wellbeing for partners of people with an amputation.
The present research aimed to determine if partners' involvement in the rehabilitative care of people who have undergone an amputation predicted their wellbeing using a quantitative, cross-sectional design. Partners of people with an amputation were contacted through amputation-related advocacy groups and asked to complete an online questionnaire. Full data were collected from 66 participants who completed demographic information and measures of perceived social support, involvement in partner's rehabilitative care, depression and posttraumatic growth. Predictors into the two regression models were chosen based on their zero-order correlations with the outcome variables. Only time since amputation and degree of prosthesis use were correlated with posttraumatic growth. Time since amputation, time in relationship and partner's involvement in rehabilitative care were negatively correlated with depression. Both models (posttraumatic growth and depression) were significant. Involvement in partner's care was an independent significant predictor for depression. More involvement in partners' rehabilitative care was a significant predictor of less depression for partners of people with an amputation. The present research provides tentative evidence in support of the planned inclusion of partners by amputation rehabilitation services. Future research is needed to determine how robust this finding is.


Effects of Shengjiangxiexin decoction on irinotecan-induced toxicity in patients with UGT1A1*28 and UGT1A1*6 polymorphisms.
To evaluate the efficacy of Shengjiangxiexin decoction (SXD), prepared with a formula from Traditional Chinese Medicine (TCM), in reducing irinotecan-induced hematological and gastrointestinal toxicities in patients with UDP-glucuronosyltransferase (UGT)1A1*28 and UGT1A1*6 polymorphisms. This clinical trial included 115 patients receiving irinotecan combined with 5-fluorouracil plus l-leucovorin (FOLFIRI) treatment. All patients consented to UGT1A1*28 and *6 gene polymorphism detection prior to chemotherapy. SXD were administered from 1 day prior to chemotherapy to 6 day post chemotherapy. Chemotherapy induced adverse reactions (neutropenia, diarrhea, nausea, vomiting, anorexia and infection) were recorded, and short-term effect of chemotherapy was evaluated regularly. A total of 50 patients had *1/*1 wild genotype, 58 patients had single allele variants with genotype *1/*6 or *1/*28 , and 7 patients had two alleles variants with genotype *6/*6, *28/*28 or *6/* 28. In *1/*6 or *1/*28 patients (high risk group), 9 patients (15.5% ) developed Ⅰ-Ⅱ grade diarrhea and no patient developed severe diarrhea; neutropenia occurred in 19 patients (32.8%) and only 3 patients (8.6% ) developed sever neutropenia. There were no significant differences in any toxic effects (neutropenia, diarrhea, nausea, vomiting, anorexia or infection) between *6 or *28 variant patients (high risk group) and wild type patients. No sever toxicity was found in high risk two alleles variants patients (*6/*6, *6/*28 or *28/*28). No significant differences were observed between UGT1A1*6/*28 polymorphisms and clinical response of chemotherapy. SXD could significantly reduce irinotecan-induced hematological and gastrointestinal toxicities in UGT1A1*28 or *6 variant patients (high risk group), while this treatment didn't affect clinical response of chemotherapy.


Many M©B(n) boron wheels are local, but not global minima.
Twenty-six planar boron wheels with a central hypercoordinate atom (M©B(n), M is a 2nd or 3rd period element) were designed following the Schleyer-Boldyrev concept of geometric and electronic fit whereby in-plane σ- as well as π-aromaticity contribute to the chemical bonding. Global minimum searches using an efficient newly implemented method reveal that most of these boron wheels are only local, rather than global minima. However, the Be©B(8) triplet planar wheel global minimum is a new member of the planar hypercoordinate M©B(n) family. Six categories classify the structures of the other global minima: planar wheels, planar non-wheel forms, quasi-two-center-wheels, as well as leaf-like, pyramid-like, and umbrella-like geometries.


Nested polymerase chain reaction strategy simultaneously targeting DNA sequences of multiple bacterial species in inflammatory joint diseases. II. Examination of sacroiliac and knee joint biopsies of patients with spondyloarthropathies and other arthritides.
Bacteria play a crucial pathogenetic role in reactive arthritis (ReA) and other forms of spondyloarthropathy (SpA) and in Lyme arthritis. Although there is evidence of local persistence of bacterial antigens no definitive method revealing microbes in peripheral joints has been established. We detected DNA of individual bacteria in synovial material by PCR. Applying molecular technology we screened simultaneously for 8 bacterial genomes in arthritis and sacroiliitis. Sacroiliac (SI) biopsy specimens taken from the SI joint of 8 patients with ankylosing spondylitis (AS, n = 5) and undifferentiated SpA (uSpA, n = 3) by computed tomography guided biopsy were investigated for presence of bacterial DNA. Similarly, synovial membrane samples obtained by office arthroscopy from 15 patients with ReA (n = 5), uSpA (n = 3), undifferentiated oligoarthritis (uOligo, n = 3), and rheumatoid arthritis (RA, n = 4) were screened. Nested PCR was performed for DNA of the following bacteria: Chlamydia trachomatis, C. pneumoniae, Yersinia enterocolitica, Salmonella enteritidis, Campylobacter jejuni, Shigella flexneri, Klebsiella pneumoniae, and Borrelia burgdorferi. No bacterial DNA was found in the SI biopsies of patients with uSpA and AS. DNA of B. burgdorferi (n = 2) and C. trachomatis (n = 1) was detected in 3 patients with uOligo, but not in patients with ReA or RA. DNA of other microbes including K. pneumoniae was not found. Patients' mean disease duration was 5.3 years (2 mo-8.4 yrs). We found bacterial DNA in peripheral joints of patients with uOligo, while in patients with ReA, AS, and uSpA no bacterial DNA was detected in peripheral or SI joints. The failure to detect bacterial DNA in patients with SpA suggests autoimmune mechanisms operate in later stages of disease.


Distinct but critical roles for integrin alphaIIbbeta3 in platelet lamellipodia formation on fibrinogen, collagen-related peptide and thrombin.
Integrins are the major receptor type known to facilitate cell adhesion and lamellipodia formation on extracellular matrix proteins. However, collagen-related peptide and thrombin have recently been shown to mediate platelet lamellipodia formation when presented as immobilized surfaces. The aims of this study were to establish if there exists a role for the platelet integrin alpha(IIb)beta(3) in this response; and if so, whether signalling from the integrin is required for lamellipodia formation on these surfaces. Real-time analysis was used to compare platelet morphological changes on surfaces of fibrinogen, collagen-related peptide or thrombin in the presence of various pharmacological inhibitors and platelets from 'knockout' mice. We demonstrate that collagen-related peptide and thrombin stimulate distinct patterns of platelet lamellipodia formation and elevation of intracellular Ca(2+) to that induced by the integrin alpha(IIb)beta(3) ligand, fibrinogen. Nevertheless, lamellipodia formation on collagen-related peptide and thrombin is dependent upon engagement of alpha(IIb)beta(3), consistent with release of alpha(IIb)beta(3) ligand(s) from platelet granules. However, the requirement for signalling by the integrin on fibrinogen can be bypassed by the addition of thrombin to the solution. These observations reveal a critical role for alpha(IIb)beta(3) in forming lamellipodia on collagen-related peptide and thrombin which is dependent on its ability to function as an adhesive receptor but not necessarily on its ability to signal. These results suggest that integrins may play an important role in lamellipodia formation triggered by nonintegrin ligands in platelets and possibly in other cell types.


The effect of insulin on expression level of nucleoside transporters in diabetic rats.
Evidence that the time course of insulin-induced changes in adenosine level in diabetic rats is different from that observed for expression of adenosine kinase prompted us to study the insulin effect on expression of nucleoside transporters in tissues of diabetic rats. RNase protection assay demonstrated that mRNA levels of equilibrative (rENT) and Na+-dependent nucleoside transporters (rCNT) were altered in diabetic tissues. The rENT1 mRNA level with respect to values obtained in age- and sex-matched nondiabetic rats was decreased by 45, 32, and 10% in diabetic heart, liver, and kidney, respectively. The level of rENT2 mRNA was lowered by 40% in diabetic kidney and heart, and by 24% in diabetic liver. Changes in the expression pattern of rCNT1 and rCNT2 in diabetic tissues differed significantly from that observed for rENT. The levels of rCNT1 and rCNT2 mRNA did not change significantly in diabetic kidney. In diabetic heart, the mRNA levels of rCNT1 and rCNT2 increased 1.7- and 2-fold, respectively. Changes in expression of nucleoside transporters were accompanied by alterations in adenosine content. Administration of insulin to diabetic rats resulted in a drop in adenosine concentration in examined tissues and return of the rCNT1, rCNT2, and rENT2 but not rENT1 mRNA levels to values observed in nondiabetic rats. In summary, these data demonstrate that insulin affects expression of nucleoside transporters in a cell-specific manner. We conclude that change in the expression level of the nucleoside transporters occurring in tissues of diabetic rat is an important factor influencing adenosine levels in the cell.


Isolation and characterization of Acidobacterium ailaaui sp. nov., a novel member of Acidobacteria subdivision 1, from a geothermally heated Hawaiian microbial mat.
A novel member of Acidobacteria was isolated from a microbial mat growing on a geothermally heated dead tree trunk in Hawai'i Volcanoes National Park (HI, USA). The rod-shaped, Gram-negative capsulated cells of strain PMMR2T were non-motile and catalase and oxidase negative. Growth occurred aerobically from 15 to 55 °C (optimum, 40 °C) and at pH values from 4.5 to 7.0 (optimum, 6.5). A limited range of sugars and organic acids supported growth. However, results of a genomic analysis suggested that various polysaccharides might be hydrolysed as carbon sources, and evidence for pectin degradation was observed in liquid cultures. A genomic analysis also revealed genes for a Group 1f uptake hydrogenase; assays with liquid cultures confirmed hydrogen consumption, including uptake at sub-atmospheric concentrations. Nitrate was not dissimilated to nitrite. Major membrane fatty acids included iso-C15 : 0 and iso-C17 : 0. The G+C content was 57.2mol%. A comparative genome analysis revealed an average nucleotide identity of 72.2 % between PMMR2T and its nearest cultured phylogenetic neighbour, Acidobacterium capsulatum ATCC 51196T (=JCM 7670T); analysis of the 16S rRNA gene revealed a 96.8 % sequence identity with Acidobacterium capsulatum ATCC 51196T. These results and other phenotypic differences indicated that strain PMMR2T represents a novel species in the genus Acidobacterium, for which the name Acidobacterium ailaaui sp. nov. is proposed. The type strain, PMMR2T (=DSM 27394T=LMG 28340T), is the second formal addition to the genus Acidobacterium.


Pre-Implantation Balloon Aortic Valvuloplasty and Clinical Outcomes Following Transcatheter Aortic Valve Implantation: A Propensity Score Analysis of the UK Registry.
Aortic valve predilation with balloon aortic valvuloplasty (BAV) is recommended before transcatheter aortic valve implantation (TAVI), despite limited data around the requirement of this preprocedural step and the potential risks of embolization. This study aimed to investigate the trends in practice and associations of BAV on short-term outcomes in the UK TAVI registry. Eleven clinical endpoints were investigated, including 30-day mortality, myocardial infarction, aortic regurgitation, valve dysfunction, and composite early safety. All endpoints were defined as per the VARC-2 definitions. Odd ratios of each endpoint were estimated using logistic regression, with data analyzed in balloon- and self-expandable valve subgroups. Propensity scores were calculated using patient demographics and procedural variables, which were included in the models of each endpoint to adjust for measured confounding. Between 2007 and 2014, 5887 patients met the study inclusion criteria, 1421 (24.1%) of whom had no BAV before TAVI valve deployment. We observed heterogeneity in the use of BAV nationally, both temporally and by center experience; rates of BAV in pre-TAVI workup varied between 30% and 97% across TAVI centers. All endpoints were similar between treatment groups in SAPIEN (Edwards Lifesciences Inc., Irvine, CA) valve patients. After correction for multiple testing, none of the endpoints in CoreValve (Medtronic, Minneapolis, MN) patients were significantly different between patients with or without predilation. Performing TAVI without predilation was not associated with adverse short-term outcomes post procedure, especially when using a balloon-expandable prosthesis. Randomized trials including different valve types are required to provide conclusive evidence regarding the utility of predilation before-TAVI.


Analysis of actin filament network organization in lamellipodia by comparing experimental and simulated images.
Protrusion of lamellipodia during cell migration depends on the assembly of actin network. The assembly mechanism, based on dendritic filament branching, has been investigated in reconstituted in vitro systems, but little is known about the dynamical and structural properties of the actin network in the lamellipodia of migrating cells. The length and orientation of filaments are difficult to measure directly in either optical or electron microscopy images because of the high filament density and overlapping of individual filaments. Here, we use the non-uniformity of optical images of the lamellipodia to extract information about the structural and dynamical properties of the underlying actin network. To determine the relationship between the image features and the properties of the network, we performed simulations of actin network assembly, based on the hypothesis of stochastic branching and capping of filaments, and produced computed ;fluorescence' and ;electron microscopy' images of the simulated network. By varying simulation parameters, in particular the actin filament density, length and orientation, we closely reproduced the contrast and the characteristic diagonal criss-cross pattern observed in the experimental optical images. Thus, matching the images of the simulated network to the experimental images allowed us to estimate parameters of actin filament network in lamellipodia.


Sequential amino acid exchange across b(0,+)-like system in chicken brush border jejunum.
In the small intestine, cationic amino acids are transported by y(+)-like and b(0,+)-like systems present in the luminal side of the epithelium. Here, we report the characterization of a b(0,+)-like system in the apical membrane of the chicken jejunum, and its properties as an amino acid exchanger. Analysis of the brush border membrane by Western blot points out the presence of rBAT (protein related to b0,+ amino acid transport system) in these membranes. A functional mechanism for amino acid exchange across this system was established by kinetic analysis measuring fluxes at varying substrate concentrations both in internal (in) and external (out) vesicle compartments. This intestinal b(0,+)-like system functions for L-arginine as an obligatory exchanger since its transport capacity increases 100-200 fold in exchange conditions, thus suggesting an important role in the intestinal absorption of cationic amino acids. The kinetic analysis of Argin efflux velocities is compatible with the formation of a ternary complex and excludes a model involving a ping-pong mechanism. The binding affinity of Argout is higher than that of Argin, suggesting a possible order of binding (Argout first) for the formation of the ternary complex during the exchange cycle. A model of double translocation pathways with alternating access is discussed.


Evaluating feasibility and acceptability of a group WHO trans-diagnostic intervention for women with common mental disorders in rural Pakistan: a cluster randomised controlled feasibility trial.
The aim of this feasibility trial was to evaluate the feasibility and acceptability of the locally adapted Group Problem Management Plus (PM+) intervention for women in the conflict affected settings in Swat, Pakistan. This mixed-methods study incorporated a quantitative component consisting of a two arm cluster randomised controlled feasibility trial, and qualitative evaluation of the acceptability of the Group PM+ to a range of stakeholder groups. For the quantitative component, on average from each of the 20 Lady Health Workers (LHWs) catchment area (20 clusters), six women were screened and recruited for the trial with score of >2 on the General Health Questionnaire and score of >16 on the WHO Disability Assessment Schedule. These LHW clusters were randomised on a 1 : 1 allocation ratio using a computer-based software through a simple randomisation method to the Group PM+ intervention or Enhanced Usual Care. The Group PM+ intervention consisted of five weekly sessions of 2 h duration delivered by local non-specialist females under supervision. The primary outcome was individual psychological distress, measured by levels of anxiety and depression on the Hospital Anxiety and Depression Scale at 7th week after baseline. Secondary outcomes include symptoms of depression, post-traumatic stress disorder (PTSD), general psychological profile, levels of functioning and generalised psychological distress. Intervention acceptability was explored through in-depth interviews. The results show that lay-helpers with no prior mental health experience can be trained to achieve the desired competency to successfully deliver the intervention in community settings under supervision. There was a good intervention uptake, with Group PM+ considered useful by participants, their families and lay-helpers. The outcome evaluation, which was not based on a large enough study to identify statistically significant results, indicated statistically significant improvements in depression, anxiety, general psychological profile and functioning. The PTSD symptoms and depressive disorder scores showed a trend in favour of the intervention. This trial showed robust acceptance in the local settings with delivery by non-specialists under supervision by local trained females. The trial paves the way for further adaptation and exploration of the outcomes through larger-scale implementation and definitive randomised controlled trials in the local settings.


Separate and concurrent symbolic predictions of sound features are processed differently.
The studies investigated the impact of predictive visual information about the pitch and location of a forthcoming sound on the sound processing. In Symbol-to-Sound matching paradigms, symbols induced predictions of particular sounds. The brain's error signals (IR and N2b components of the event-related potential) were measured in response to occasional violations of the prediction, i.e., when a sound was incongruent to the corresponding symbol. IR and N2b index the detection of prediction violations at different levels, IR at a sensory and N2b at a cognitive level. Participants evaluated the congruency between prediction and actual sound by button press. When the prediction referred to only the pitch or only the location feature (Experiment 1), the violation of each feature elicited IR and N2b. The IRs to pitch and location violations revealed differences in the in time course and topography, suggesting that they were generated in feature-specific sensory areas. When the prediction referred to both features concurrently (Experiment 2), that is, the symbol predicted the sound's pitch and location, either one or both predictions were violated. Unexpectedly, no significant effects in the IR range were obtained. However, N2b was elicited in response to all violations. N2b in response to concurrent violations of pitch and location had a shorter latency. We conclude that associative predictions can be established by arbitrary rule-based symbols and for different sound features, and that concurrent violations are processed in parallel. In complex situations as in Experiment 2, capacity limitations appear to affect processing in a hierarchical manner. While predictions were presumably not reliably established at sensory levels (absence of IR), they were established at more cognitive levels, where sounds are represented categorially (presence of N2b).


Effect of end-stage hip, knee, and ankle osteoarthritis on walking mechanics.
This study tested the hypothesis that the presence of isolated ankle (A-OA; N=30), knee (K-OA; N=20), or hip (H-OA; N=30) osteoarthritis (OA) compared to asymptomatic controls (N=15) would lead to mechanical changes in the affected joint but also in all other lower limb joints and gait overall. Stride length, stance and swing times, as well as joint angles and moments at the hip, knee, and ankle were derived from 3-D kinematic and kinetic data collected from seven self-selected speed walking trial. Values were compared across groups using a 1×4 ANCOVA, covarying for walking speed. With walking speed controlled, the results indicated a reduction in hip and knee extension and ankle plantar flexion in accordance with the joint affected. In addition, OA in one joint had strong effects on other joints. In both H-OA and K-OA groups the hip never passed into extension, and A-OA subjects significantly changed hip kinematics to compensate for lack of plantar flexion. Finally, OA in any joint led to lower peak vertical forces as well as extension and plantar flexion moments compared to controls. The presence of end-stage OA at various lower extremity joints results in compensatory gait mechanics that cause movement alterations throughout the lower extremity. This work reinforces our understanding of the complex interaction of joints of the lower limb and the importance of focusing on the mechanics of the entire lower limb when considering gait disability and potential interventions in patients with isolated OA.


Localization of the coactivator Cdh1 and the cullin subunit Apc2 in a cryo-electron microscopy model of vertebrate APC/C.
The anaphase-promoting complex/cyclosome (APC/C) is a ubiquitin ligase with essential functions in mitosis, meiosis, and G1 phase of the cell cycle. APC/C recognizes substrates via coactivator proteins such as Cdh1, and bound substrates are ubiquitinated by E2 enzymes that interact with a hetero-dimer of the RING subunit Apc11 and the cullin Apc2. We have obtained three-dimensional (3D) models of human and Xenopus APC/C by angular reconstitution and random conical tilt (RCT) analyses of negatively stained cryo-electron microscopy (cryo-EM) preparations, have determined the masses of these particles by scanning transmission electron microscopy (STEM), and have mapped the locations of Cdh1 and Apc2. These proteins are located on the same side of the asymmetric APC/C, implying that this is where substrates are ubiquitinated. We have further identified a large flexible domain in APC/C that adopts a different orientation upon Cdh1 binding. Cdh1 may thus activate APC/C both by recruiting substrates and by inducing conformational changes.


The translation of an evidence-based preschool physical activity intervention from in-person to online delivery of professional development to preschool teachers.
Study of Health and Activity in Preschool Environments (SHAPES) aimed to increase preschoolers' physical activity by engaging preschool teachers in professional development designed to modify instructional practices. SHAPES originated as a randomized controlled trial, tested in 16 preschools and disseminated to 4 control schools, and was shown to be effective in increasing preschool children's physical activity (Phase I, 2008-2012). This article describes the steps taken to translate an evidence-based physical activity intervention, SHAPES, from in-person delivery to online delivery of professional development to preschool teachers. In Phase II (2013-2016), professional development delivery was modified from an in-person to an online program for preschool teachers. Teacher implementation completeness and fidelity were examined in 10 pilot preschools and then replicated in 30 preschools. Large-scale dissemination of SHAPES began in Phase III (2017-2019), reaching 818 teachers from 228 unique centers after 2 years. The final SHAPES program consisted of six online modules, an accompanying guidebook, over 90 activities, self-assessment techniques, a video library, and an electronic community forum. The SHAPES core elements, defined in Phase I, were retained throughout all three phases. The Phase II change to online delivery was characterized by high levels of implementation completeness and fidelity. Results of Phase III statewide dissemination revealed comparable implementation completeness and fidelity. An intervention shown to be effective in a randomized controlled trial can be translated into an online professional development program and disseminated on a large scale in a timely manner.


Multimodal Investigations of Reward Circuitry and Anhedonia in Adolescent Depression.
Depression is a highly prevalent condition with devastating personal and public health consequences that often first manifests during adolescence. Though extensively studied, the pathogenesis of depression remains poorly understood, and efforts to stratify risks and identify optimal interventions have proceeded slowly. A major impediment has been the reliance on an all-or-nothing categorical diagnostic scheme based solely on whether a patient endorses an arbitrary number of common symptoms for a sufficiently long period. This approach masks the well-documented heterogeneity of depression, a disorder that is highly variable in presentation, severity, and course between individuals and is frequently comorbid with other psychiatric conditions. In this targeted review, we outline the limitations of traditional diagnosis-based research and instead advocate an alternative approach centered around symptoms as unique dimensions of clinical dysfunction that span across disorders and more closely reflect underlying neurobiological abnormalities. In particular, we highlight anhedonia-the reduced ability to anticipate and experience pleasure-as a specific, quantifiable index of reward dysfunction and an ideal candidate for dimensional investigation. Anhedonia is a core symptom of depression but also a salient feature of numerous other conditions, and its severity varies widely within clinical and even healthy populations. Similarly, reward dysfunction is a hallmark of depression but is evident across many psychiatric conditions. Reward function is especially relevant in adolescence, a period characterized by exaggerated reward-seeking behaviors and rapid maturation of neural reward circuitry. We detail extensive work by our research group and others to investigate the neural and systemic factors contributing to reward dysfunction in youth, including our cumulative findings using multiple neuroimaging and immunological measures to study depressed adolescents but also trans-diagnostic cohorts with diverse psychiatric symptoms. We describe convergent evidence that reward dysfunction: (a) predicts worse clinical outcomes, (b) is associated with functional and chemical abnormalities within and beyond the neural reward circuitry, (c) is linked to elevated peripheral levels of inflammatory biomarkers, and (d) manifests early in the course of illness. Emphasis is placed on high-resolution neuroimaging techniques, comprehensive immunological assays, and data-driven analyses to fully capture and characterize the complex, interconnected nature of these systems and their contributions to adolescent reward dysfunction.


Mortality from different causes associated with meat, heme iron, nitrates, and nitrites in the NIH-AARP Diet and Health Study: population based cohort study.
Objective To determine the association of different types of meat intake and meat associated compounds with overall and cause specific mortality.Design Population based cohort study.Setting Baseline dietary data of the NIH-AARP Diet and Health Study (prospective cohort of the general population from six states and two metropolitan areas in the US) and 16 year follow-up data until 31 December 2011.Participants 536 969 AARP members aged 50-71 at baseline.Exposures Intake of total meat, processed and unprocessed red meat (beef, lamb, and pork) and white meat (poultry and fish), heme iron, and nitrate/nitrite from processed meat based on dietary questionnaire. Adjusted Cox proportional hazards regression models were used with the lowest fifth of calorie adjusted intakes as reference categories.Main outcome measure Mortality from any cause during follow-up.Results An increased risk of all cause mortality (hazard ratio for highest versus lowest fifth 1.26, 95% confidence interval 1.23 to 1.29) and death due to nine different causes associated with red meat intake was observed. Both processed and unprocessed red meat intakes were associated with all cause and cause specific mortality. Heme iron and processed meat nitrate/nitrite were independently associated with increased risk of all cause and cause specific mortality. Mediation models estimated that the increased mortality associated with processed red meat was influenced by nitrate intake (37.0-72.0%) and to a lesser degree by heme iron (20.9-24.1%). When the total meat intake was constant, the highest fifth of white meat intake was associated with a 25% reduction in risk of all cause mortality compared with the lowest intake level. Almost all causes of death showed an inverse association with white meat intake.Conclusions The results show increased risks of all cause mortality and death due to nine different causes associated with both processed and unprocessed red meat, accounted for, in part, by heme iron and nitrate/nitrite from processed meat. They also show reduced risks associated with substituting white meat, particularly unprocessed white meat.


Treatment options for nosocomial pneumonia due to MRSA.
Nosocomial pneumonia, which includes hospital-acquired pneumonia, ventilator-associated pneumonia, and health care associated pneumonia, remains an important cause of morbidity and mortality. The continuing emergence of methicillin-resistant Staphylococcus aureus (MRSA) as a nosocomial pneumonia pathogen is particularly problematic not only because of its prevalence, but also because antimicrobial resistance is increasingly associated with inappropriate empirical antibiotic therapy. As a result, intensivists are faced with the dual goals of providing initial accurate broad-spectrum antibiotic coverage to reduce mortality while minimizing the risk for the emergence of antimicrobial resistance. These competing goals can be achieved by using an approach that initially delivers liberal broad-spectrum coverage followed by de-escalation once culture results and serial clinical observations become available. In ventilator-associated pneumonia, linezolid has demonstrated favorable activity against Gram-positive bacteria, including MRSA, and is recommended in evidence-based guidelines as an alternative to vancomycin, particularly when MRSA is documented as the etiology.


Renal effects of indoprofen in rhesus monkeys.
The non-steroidal anti-inflammatory analgesic alpha-[4-(1-oxo-2-iso-indolinyl)-phenyl]propionic acid (indoprofen) was given p.o. in gelatin capsules to six groups of six Rhesus monkeys (three males and three females/group) for 15 months. Group I (five males and five females) served as control and was given empty gelatin capsules. The following daily doses were used: Group 2--20 mg/kg b.i.d.; Group 3--40 mg/kg q.d.; Group 4--30 mg/kg b.i.d.; Group 5--60 mg/kg q.d.; Group 6--40 mg/kg b.i.d. and Group 7--80 mg/kg q.d. Except for some elevated blood urea nitrogen levels after 12 months in the animals of Group 7, no changes in clinical function test values, absolute and relative kidney weights or renal gross abnormalities were found. The renal papillae and pelves were normal and not affected in all monkeys. The microscopic examination revealed a focal chronic interestitial nephritis in some of the monkeys. In the survivors the incidence of this disease increased from one out of ten control animals to three out of five monkeys in Group 7 (80 mg/kg/d). However, this quantal, statistically significant difference became biologically irrelevant and no longer statistically significant when these microscopic changes were quantified. The most severe case was seen in the control animal and the least pronounced in monkeys having received 80 mg/kg/d. When these cases of interstitial nephritis were further evaluated using a multifactorial point sampling procedure, no significant quantitative changes in renal tissue components, indicative of analgesic-induced nephropathy, were detectable.


Oral Health, Obesity Status and Nutritional Habits in Turkish Children and Adolescents: An Epidemiological Study.
Studies evaluating the relationship between oral health status and obesity have provided conflicting data. Therefore, there is a great need to investigate and clarify the possible connection in a comprehensive sample. To assess the relationship of obesity and oral health status among children and adolescents aged 6 to 17 years-old. Cross-sectional study. Data were obtained from 4,534 children and adolescents (2,018 boys and 2,516 girls). Questionnaires were sent home prior to examination; afterwards, anthropometric and dental data were collected from participants. Community Periodontal Index (CPI) and number of decayed, missing, and filled teeth in the permanent dentition (DMFT), and deciduous dentition (dmft) index were used to measure oral health status. Height, body weight, body mass index (BMI), waist circumference (WC), and body fat percentage were analyzed. For DMFT scores, healthy (score=0) girls and boys had significantly higher BMI and WC values than unhealthy (score>1) girls and boys (p<0.05). Healthy girls had higher fat percentage values than unhealthy girls (p<0.05). In terms of CPI scores, healthy boys had lower BMI and WC values than unhealthy boys (p<0.05). According to multiple binary logistic regression results for model 1, BMI predicted DMFT scores in both genders but CPI scores only in boys. No beverage consumption predicted DMFT scores in boys, while milk consumption predicted DMFT scores in girls. No meal skipping predicted CPI scores in boys. For model 2, WC predicted DMFT scores in both genders and CPI scores only in boys. Milk consumption predicted DMFT scores only in girls. No meal skipping predicted CPI scores for both gender (p<0.05). According to DMFT, there were significant differences between the frequencies of the BMI groups (normal weight, overweight and obese) at the age of 7 (girls only), 9, 10, and 16 (boys only) years and overall (only girls) (p<0.05). According to CPI, significant differences between the frequencies of the BMI groups at the age of 16 (boys only) and 17 (girls only) were seen (p<0.05). Periodontal and dental status appears to correlate with nutritional habits and obesity. Obesity and dental/periodontal diseases are multifactorial diseases that follow similar risk patterns and develop from an interaction between chronic conditions originating early in life. It is important for all health professionals to educate patients at risk about the progression of periodontal and dental diseases and the importance of proper oral hygiene.


OMERACT Rheumatoid Arthritis Magnetic Resonance Imaging Studies. Exercise 5: an international multicenter reliability study using computerized MRI erosion volume measurements.
Scoring erosions on magnetic resonance imaging (MRI) is one method of estimating damage in patients with rheumatoid arthritis (RA), but it has limitations. The aim of this pilot study was to assess the feasibility and inter-reader reliability of computer assisted erosion volume estimation in patients with RA. Intra-reader and inter-occasion reliability was also assessed, and different slice thicknesses were compared in terms of erosion volume estimation. A 3 mm slice thickness 3D gradient-echo sequence followed by a 1 mm sequence was performed at baseline and repeated within 24 h with metacarpophalangeal (MCP) joints 2 to 5 of the dominant hand included in the field of view. Three readers were instructed to grade MCP 2 and 3 using the OMERACT grading system and then to measure the erosion volume of the same joints using OSIRIS software. The inter-reader reliability of the grading method and the volume method was calculated, as well as the inter-occasion reliability, by comparing results from each reader from baseline to the followup scan. One reader performed repeat volume measurements on 5 patients to assess the intra-reader reliability. Five patients were included in the study. Expressed in terms of intraclass correlation coefficients (ICC), the inter-reader and inter-occasion reliability of the volume method were comparable to the existing OMERACT scoring system, but large systematic differences in volume estimations were found between readers. The intra-reader reliability was excellent. Good correlation was demonstrated between the total erosion scores and the total erosion volumes. For both erosion volumes and erosion scores, 1 mm and 3 mm acquisitions produced variable results between readers, with no clear pattern of underestimation or overestimation for either slice thickness. The volume estimation method was more time consuming, taking roughly 5 times as long as the scoring method. Computerized MRI erosion volume measurements are feasible, with high intra-observer and inter-occasion reliabilities. Despite high ICC, the inter-observer reliability is not sufficient for multicenter use without prior reader training and calibration. The optimal slice thickness was not determined.


Comparative single-cell genomics reveals potential ecological niches for the freshwater acI Actinobacteria lineage.
Members of the acI lineage of Actinobacteria are the most abundant microorganisms in most freshwater lakes; however, our understanding of the keys to their success and their role in carbon and nutrient cycling in freshwater systems has been hampered by the lack of pure cultures and genomes. We obtained draft genome assemblies from 11 single cells representing three acI tribes (acI-A1, acI-A7, acI-B1) from four temperate lakes in the United States and Europe. Comparative analysis of acI SAGs and other available freshwater bacterial genomes showed that acI has more gene content directed toward carbohydrate acquisition as compared to Polynucleobacter and LD12 Alphaproteobacteria, which seem to specialize more on carboxylic acids. The acI genomes contain actinorhodopsin as well as some genes involved in anaplerotic carbon fixation indicating the capacity to supplement their known heterotrophic lifestyle. Genome-level differences between the acI-A and acI-B clades suggest specialization at the clade level for carbon substrate acquisition. Overall, the acI genomes appear to be highly streamlined versions of Actinobacteria that include some genes allowing it to take advantage of sunlight and N-rich organic compounds such as polyamines, di- and oligopeptides, branched-chain amino acids and cyanophycin. This work significantly expands the known metabolic potential of the cosmopolitan freshwater acI lineage and its ecological and genetic traits.


Advancements towards the implementation of clinical phase-contrast breast computed tomography at Elettra.
Breast computed tomography (BCT) is an emerging application of X-ray tomography in radiological practice. A few clinical prototypes are under evaluation in hospitals and new systems are under development aiming at improving spatial and contrast resolution and reducing delivered dose. At the same time, synchrotron-radiation phase-contrast mammography has been demonstrated to offer substantial advantages when compared with conventional mammography. At Elettra, the Italian synchrotron radiation facility, a clinical program of phase-contrast BCT based on the free-space propagation approach is under development. In this paper, full-volume breast samples imaged with a beam energy of 32 keV delivering a mean glandular dose of 5 mGy are presented. The whole acquisition setup mimics a clinical study in order to evaluate its feasibility in terms of acquisition time and image quality. Acquisitions are performed using a high-resolution CdTe photon-counting detector and the projection data are processed via a phase-retrieval algorithm. Tomographic reconstructions are compared with conventional mammographic images acquired prior to surgery and with histologic examinations. Results indicate that BCT with monochromatic beam and free-space propagation phase-contrast imaging provide relevant three-dimensional insights of breast morphology at clinically acceptable doses and scan times.


Abrupt rise of end tidal carbon dioxide level was a specific but non-sensitive marker of return of spontaneous circulation in patient with out-of-hospital cardiac arrest.
To evaluate the diagnostic accuracy of an abrupt and sustained increase in end-tidal carbon dioxide (ETCO2) to indicate return of spontaneous circulation (ROSC) during resuscitation of patient with out-of-hospital cardiac arrest. Cross-sectional study. Emergency department of two regional hospitals. Patients with age ≥18 years old, suffered non-traumatic out-of-hospital cardiac arrest with active resuscitation and endotracheal intubation performed in emergency department, were included. ETCO2 value was charted throughout resuscitation. Time of ROSC was remarked. ETCO2 levels before and after ROSC were compared. Diagnostic accuracy of ETCO2 rise ≥10mmHg, ETCO2 rise ≥20mmHg, and ETCO2 rise to the level ≥40mmHg were evaluated for indicating ROSC. ETCO2 level immediately after ROSC was higher as compared to the value before return of circulation (median ETCO2 was 32mmHg and 41mmHg respectively, p=0.033). With ETCO2 rise ≥10mmHg, the sensitivity was low (33%, 95% CI 22-47%), while specificity was 97% (95% CI 91-99%). Positive and negative predictive values were 83% (95% CI 62-95%) and 74% (95% CI 66-81%) respectively. The diagnostic accuracy was higher in cardiac arrest with presumed non-cardiac etiology (sensitivity 45%, specificity 100%) as compared to those with cardiac etiology (sensitivity 18%, specificity 97%). The feature of an abrupt rise of ETCO2 was a specific but non-sensitive marker of ROSC in patient with out-of-hospital cardiac arrest.


The contributions of metabolomics in the discovery of new therapeutic targets in Alzheimer's disease.
Alzheimer's disease (AD) leads to the progressive loss of memory and other cognitive functions. It is the most common form of dementia in the elderly and has become a major public health problem due to the increase in life expectancy. Although the detection of AD is based on several neuropsychological tests, imaging, and biological analyses, none of these biomarkers allows a clear understanding of the pathophysiological mechanisms involved in the disease, and no efficient treatment is currently available. Metabolomics, which allows the study of biochemical alterations underlying pathological processes, could help to identify these mechanisms, to discover new therapeutic targets, and to monitor the therapeutic response and disease progression. In this review, we have summarized and analyzed the results from a number of studies on metabolomics analyses performed in biological samples originated from the central nervous system, in AD subjects, and in animal models of this disease. This synthesis revealed modified expression of specific metabolites in pathological conditions which allowed the identification of significantly impacted metabolic pathways both in animals and humans, such as the arginine biosynthesis and the alanine, aspartate, and glutamate metabolism. We discuss the potential biochemical mechanisms involved, the extent to which they could impact the specific hallmarks of AD, and the therapeutic approaches which could be proposed as a result.


Clinical Overlap of Multiple Sclerosis and Autoimmune Hepatitis: Three Cases.
Multiple sclerosis (MS) is an autoimmune, inflammatory disease characterized by demyelination and axonal degeneration in the central nervous system. MS is the second major cause of disability following trauma, and is mostly seen between the ages of 20 - 40 years and in women. Autoimmune hepatitis (AH) is a chronic disease characterized by hypergammaglobulinemia, high levels of transaminases, presence of antibodies, and histologically by the necroinflammatory process with interface hepatitis. In AH, the etiological agent of the disease and the cause of liver injury remain unknown. MS may be associated with AH, autoimmune thyroiditis, and type 1 diabetes mellitus (DM). In literature, 8 cases with overlap of MS and AH have been reported. In this report, we present 3 cases which were detected with overlap of MS and AH, and are very rare condition in literature.


Rising consumption of meat and milk in developing countries has created a new food revolution.
People in developing countries currently consume on average one-third the meat and one-quarter of the milk products per capita compared to the richer North, but this is changing rapidly. The amount of meat consumed in developing countries over the past has grown three times as much as it did in the developed countries. The Livestock Revolution is primarily driven by demand. Poor people everywhere are eating more animal products as their incomes rise above poverty level and as they become urbanized. By 2020, the share of developing countries in total world meat consumption will expand from 52% currently to 63%. By 2020, developing countries will consume 107 million metric tons (mmt) more meat and 177 mmt more milk than they did in 1996/1998, dwarfing developed-country increases of 19 mmt for meat and 32 mmt for milk. The projected increase in livestock production will require annual feed consumption of cereals to rise by nearly 300 mmt by 2020. Nonetheless, the inflation-adjusted prices of livestock and feed commodities are expected to fall marginally by 2020, compared to precipitous declines in the past 20 y. Structural change in the diets of billions of people is a primal force not easily reversed by governments. The incomes and nutrition of millions of rural poor in developing countries are improving. Yet in many cases these dietary changes also create serious environmental and health problems that require active policy involvement to prevent irreversible consequences.


The Use of Rhythmic Auditory Stimulation to Optimize Treadmill Training for Stroke Patients: A Randomized Controlled Trial.
The use of functional music in gait training termed rhythmic auditory stimulation (RAS) and treadmill training (TT) have both been shown to be effective in stroke patients (SP). The combination of RAS and treadmill training (RAS-TT) has not been clinically evaluated to date. The aim of the study was to evaluate the efficacy of RAS-TT on functional gait in SP. The protocol followed the design of an explorative study with a rater-blinded three arm prospective randomized controlled parallel group design. Forty-five independently walking SP with a hemiparesis of the lower limb or an unsafe and asymmetrical walking pattern were recruited. RAS-TT was carried out over 4 weeks with TT and neurodevelopmental treatment based on Bobath approach (NDT) serving as control interventions. For RAS-TT functional music was adjusted individually while walking on the treadmill. Pre and post-assessments consisted of the fast gait speed test (FGS), a gait analysis with the locometre (LOC), 3 min walking time test (3MWT), and an instrumental evaluation of balance (IEB). Raters were blinded to group assignments. An analysis of covariance (ANCOVA) was performed with affiliated measures from pre-assessment and time between stroke and start of study as covariates. Thirty-five participants (mean age 63.6 ± 8.6 years, mean time between stroke and start of study 42.1 ± 23.7 days) completed the study (11 RAS-TT, 13 TT, 11 NDT). Significant group differences occurred in the FGS for adjusted post-measures in gait velocity [F (2, 34) = 3.864, p = 0.032; partial η2 = 0.205] and cadence [F (2, 34) = 7.656, p = 0.002; partial η2 = 0.338]. Group contrasts showed significantly higher values for RAS-TT. Stride length results did not vary between the groups. LOC, 3MWT, and IEB did not indicate group differences. One patient was withdrawn from TT because of pain in one arm. The study provides first evidence for a higher efficacy of RAS-TT in comparison to the standard approaches TT and NDT in restoring functional gait in SP. The results support the implementation of functional music in neurological gait rehabilitation and its use in combination with treadmill training. Clinical Trial Registration: https://www.drks.de/drks_web/, identifier DRKS00014603.


The Ia afferent feedback of a given movement evokes the illusion of the same movement when returned to the subject via muscle tendon vibration.
The aim of the present study was to further investigate the contribution of primary muscle spindle feedback to proprioception and higher brain functions, such as movement trajectory recognition. For this purpose, complex illusory movements were evoked in subjects by applying patterns of muscle tendon vibration mimicking the natural Ia afferent pattern. Ia afferent messages were previously recorded using microneurographic method from the six main muscle groups acting on the ankle joint during imposed "writing like" movements. The mean Ia afferent pattern was calculated for each muscle group and used as a template to pilot each vibrator. Eleven different vibratory patterns were applied to ten volunteers. Subjects were asked both to copy the perceived illusory movements by hand on a digitizing tablet and to recognize and name the corresponding graphic symbol. The results show that the Ia afferent feedback of a given movement evokes the illusion of the same movement when it is applied to the subject via the appropriate pattern of muscle tendon vibration. The geometry and the kinematic parameters of the imposed and illusory movements are very similar and the so-called "two-thirds power law" is present in the reproduction of the vibration-induced illusory movements. Vibrations within the "natural" frequency range of Ia fibres firing (around 30 Hz) produce clear illusions of movements in all the tested subjects. In addition, increasing the mean frequency of the vibration patterns resulted in a linear increase in the size of the illusory movements. Lastly, the subjects were able to recognize and name the symbols evoked by the vibration-induced primary muscle spindle afferent patterns in 83% of the trials. These findings suggest that the "proprioceptive signature" of a given movement is associated with the corresponding "perceptual signature". The neural mechanisms possibly underlying the sensory to perceptual transformation are discussed in the general framework of "the neuronal population vector model".


ESMO consensus conference on malignant lymphoma: general perspectives and recommendations for prognostic tools in mature B-cell lymphomas and chronic lymphocytic leukaemia.
The European Society for Medical Oncology (ESMO) consensus conference on mature B-cell lymphomas and chronic lymphocytic leukaemia (CLL) was held on 20 June 2015 in Lugano, Switzerland, and included a multidisciplinary panel of 25 leading experts. The aim of the conference was to develop recommendations on critical subjects difficult to consider in detail in the ESMO Clinical Practice Guidelines. The following areas were identified: (i) the elderly patient, (ii) prognostic factors suitable for clinical use and (iii) the 'ultra-high-risk' group. Before the conference, the expert panel was divided into three working groups; each group focused on one of these areas in order to address four clinically relevant questions relating to that topic. All relevant scientific literature, as identified by the experts, was reviewed in advance. During the consensus conference, each working group developed recommendations to address each of the four questions assigned to their group. These recommendations were then presented to the entire panel and a consensus was reached. This manuscript presents recommendations dedicated to the second area of interest, i.e. prognostic factors suitable for clinical use. The four topics [i.e. interim positron emission tomography (PET), TP53 mutations, cell of origin (COO) and minimal residual disease (MRD)] were primarily chosen because of the bulk of available data together with the lack of clear guidance regarding their use in clinical practice and within clinical trials. Results, including a summary of evidence supporting each recommendation, are detailed in this manuscript. The panel acknowledged that detection of TP53 inactivation by deletion or mutation in CLL should be implemented in clinical practice (level of evidence I, strength of recommendation A). Due to their potentially high prognostic value, at least in some lymphoma entities, implementation of interim PET, COO and MRD was highly recommended in the context of clinical trials. All expert panel members approved this final article.


Muscle-specific PPARgamma-deficient mice develop increased adiposity and insulin resistance but respond to thiazolidinediones.
Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) by thiazolidinediones (TZDs) improves insulin resistance by increasing insulin-stimulated glucose disposal in skeletal muscle. It remains debatable whether the effect of TZDs on muscle is direct or indirect via adipose tissue. We therefore generated mice with muscle-specific PPARgamma knockout (MuPPARgammaKO) using Cre/loxP recombination. Interestingly, MuPPARgammaKO mice developed excess adiposity despite reduced dietary intake. Although insulin-stimulated glucose uptake in muscle was not impaired, MuPPARgammaKO mice had whole-body insulin resistance with a 36% reduction (P < 0.05) in the glucose infusion rate required to maintain euglycemia during hyperinsulinemic clamp, primarily due to dramatic impairment in hepatic insulin action. When placed on a high-fat diet, MuPPARgammaKO mice developed hyperinsulinemia and impaired glucose homeostasis identical to controls. Simultaneous treatment with TZD ameliorated these high fat-induced defects in MuPPARgammaKO mice to a degree identical to controls. There was also altered expression of several lipid metabolism genes in the muscle of MuPPARgammaKO mice. Thus, muscle PPARgamma is not required for the antidiabetic effects of TZDs, but has a hitherto unsuspected role for maintenance of normal adiposity, whole-body insulin sensitivity, and hepatic insulin action. The tissue crosstalk mediating these effects is perhaps due to altered lipid metabolism in muscle.