Cocoa inhibits platelet activation and function.
Epidemiologic studies have shown inverse associations between dietary polyphenols and mortality from coronary heart disease. However, the basis for this protective association is uncertain. Food polyphenols reportedly have antioxidant properties and decrease platelet function in vitro. This study sought to evaluate whether consumption of a polyphenol-rich cocoa beverage modulates human platelet activation and primary hemostasis. Peripheral blood was obtained from 30 healthy subjects before and 2 and 6 h after ingestion of a cocoa beverage (n = 10), a caffeine-containing control beverage (n = 10), or water (n = 10). Platelet activation was measured in terms of expression of activation-dependent platelet antigens and platelet microparticle formation by using fluorescent-labeled monoclonal antibodies and flow cytometry. Primary platelet-related hemostasis was measured with a platelet function analyzer. Ex vivo epinephrine- or ADP-stimulated expression of the fibrinogen-binding conformation of glycoprotein IIb-IIIa was lower 2 and 6 h after consumption of cocoa than before consumption. Cocoa consumption also decreased ADP-stimulated P-selectin expression. In contrast, epinephrine-induced platelet glycoprotein IIb-IIIa expression increased after consumption of the caffeine-containing beverage but not after water consumption. Platelet microparticle formation decreased 2 and 6 h after cocoa consumption but increased after caffeine and water consumption. Primary hemostasis in response to epinephrine in vitro was inhibited 6 h after cocoa consumption. The caffeine-containing beverage inhibited ADP-induced primary hemostasis 2 and 6 h after consumption. Cocoa consumption suppressed ADP- or epinephrine-stimulated platelet activation and platelet microparticle formation. Cocoa consumption had an aspirin-like effect on primary hemostasis.


Correlation of expression profiles between microRNAs and mRNA targets using NCI-60 data.
MicroRNAs (miRNAs) are small non-coding RNAs affecting the expression of target genes via translational repression or mRNA degradation mechanisms. With the increasing availability of mRNA and miRNA expression data, it might be possible to assess functional targets using the fact that a miRNA might down-regulate its target mRNAs. In this work we computed the correlation of expression profiles between miRNAs and target mRNAs using the NCI-60 expression data. The aim is to investigate whether the correlations between miRNA and mRNA expression profiles, either positive or negative, can be used to assist the identification of functional miRNA-mRNA relationships. Predicted miRNA-mRNA interactions were taken from TargetScan 4.1 and miRBase release 5. Pearson correlation coefficients between the miRNA and the mRNA expression profiles were computed using NCI-60 data. The correlation coefficients were then subject to the Benjamini and Hochberg correction. Our results show that the percentage of TargetScan-predicted miRNA-mRNA interactions having negative correlation in expression profiles is higher than that of miRBase-predicted pairs. Using the experimentally validated miRNA targets listed in TarBase, genes involved in mRNA degradation show more negative correlations between miRNA and mRNA expression profiles, comparing with genes involved in translational repression. Furthermore, correlation analysis for miRNAs and mRNAs transcribed from the same genes shows that correlations of expression profiles between intronic miRNAs and host genes tend to be positive. Finally we found that a target gene might be down-regulated by more than one miRNAs sharing the same seed region. Our results suggest that expression profiles can be used in the computational identification of functional miRNA-target associations. One can expect a higher chance of finding negatively correlated expression profiles for TargetScan-predicted interactions than for miRBase-predicted ones. With limited experimentally validated miRNA-target interactions, expression profiles can only serve as a supplementary role in finding interactions between miRNAs and mRNAs.


In silico characterisation and chromosomal localisation of human RRH (peropsin)--implications for opsin evolution.
The vertebrate opsins are proteins which utilise a retinaldehyde chromophore in their photosensory or photoisomerase roles in the visual/irradiance detection cycle. The majority of the opsins, such as rod and cone opsins, have a very highly conserved gene structure suggesting a common lineage. Exceptions to this are RGR-opsin and melanopsin, whose genes have very different intron insertion positions. The gene structure of another opsin, peropsin (retinal pigment epithelium-derived rhodopsin homologue, RRH) is unknown. By in silico analysis of the GenBank database we have determined that the human RRH comprises 7 exons spanning approximately 16.5 kb and is localised to chromosome 4q25 in the following gene sequence: cen-EGF-RRH-IF-qter - a position that excludes this gene as a candidate for the RP29 autosomal recessive retinitis pigmentosa locus. A comparison of opsin gene structures reveals that RRH and RGR share two common intron (introns 1 and 4) insertion positions which may reflect a shared ancestral gene. The opsins comprise a diverse group of genes which appear to have arisen from three different lineages. These lineages comprise the "classical opsin superfamily" which includes the rod and cone opsins, pinopsin, VA-opsin, parapinopsin and encephalopsin; the RRH and RGR group; and the melanopsin line. A common lineage for RRH and RGR, together with their sites of expression in the RPE, indicates that peropsin may act as a retinal isomerase.


Comparing Two Mycobacterium tuberculosis Genomes from Chinese Immigrants with Native Genomes Using Mauve Alignments.
The number of immigrants with tuberculosis (TB) increases each year in South Korea. Determining the transmission dynamics based on whole genome sequencing (WGS) to cluster the strains has been challenging. WGS, annotation refinement, and orthology assignment for the GenBank accession number acquisition were performed on two clinical isolates from Chinese immigrants. In addition, the genomes of the two isolates were compared with the genomes of Mycobacterium tuberculosis isolates, from two native Korean and five native Chinese individuals using a phylogenetic topology tree based on the Multiple Alignment of Conserved Genomic Sequence with Rearrangements (Mauve) package. The newly assigned accession numbers for two clinical isolates were CP020381.2 (a Korean-Chinese from Yanbian Province) and CP022014.1 (a Chinese from Shandong Province), respectively. Mauve alignment classified all nine TB isolates into a discriminative collinear set with matched regions. The phylogenetic analysis revealed a rooted phylogenetic tree grouping the nine strains into two lineages: strains from Chinese individuals and strains from Korean individuals. Phylogenetic trees based on the Mauve alignments were supposed to be useful in revealing the dynamics of TB transmission from immigrants in South Korea, which can provide valuable information for scaling up the TB screening policy for immigrants.


Insulin-like growth factor-I binds selectively to human peripheral blood monocytes and B-lymphocytes.
Insulin-like growth factor-I (IGF-I) is a potential modulator of responses to a variety of immunological challenges. Previous studies have suggested that specific IGF-I receptors are present on peripheral blood leukocytes, including polymorphonuclear leukocytes, lymphocytes, and monocytes. We sought to determine what type of IGF receptor was present on peripheral blood mononuclear cells and which types of cells possessed these receptors. Binding of [125I]IGF-I to mononuclear cells was inhibited by both unlabeled IGF-I and insulin, insulin being 200-fold less potent than IGF-I. Covalent affinity labeling with [125I]IGF-I, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, revealed a specific binding species with an apparent mol wt of 130 kDa. Two color flow cytometric analysis of mononuclear cells stained with mouse monoclonal antibodies specific for the human IGF-I receptor, the human insulin receptor, and monoclonal antibodies directed against specific monocyte and lymphocyte subset cell surface antigens revealed that both IGF-I receptors and insulin receptors were present on nearly all monocytes and B-lymphocytes, but were present on only 2% of T-lymphocytes. We conclude from these data that among human peripheral blood nonactivated mononuclear cells, IGF-I binds to specific type I IGF receptors found predominantly on monocytes and B-lymphocytes.


Predicting pituitary adenoma consistency with preoperative magnetic resonance elastography.
Pituitary adenoma is one of the most common primary intracranial neoplasms. Most of these tumors are soft, but up to 17% may have a firmer consistency. Therefore, knowing the tumor consistency in the preoperative setting could be helpful. Multiple imaging methods have been proposed to predict tumor consistency, but the results are controversial. This study aimed to evaluate the efficacy of MR elastography (MRE) in predicting tumor consistency and its potential use in a series of patients with pituitary adenomas. Thirty-eight patients with pituitary adenomas (≥ 2.5 cm) were prospectively evaluated with MRI and MRE before surgery. Absolute MRE stiffness values and relative MRE stiffness ratios, as well as the relative ratio of T1 signal, T2 signal, and diffusion-weighted imaging apparent diffusion coefficient (ADC) values were determined prospectively by calculating the ratio of those values in the tumor to adjacent left temporal white matter. Tumors were classified into three groups according to surgical consistency (soft, intermediate, and firm). Statistical analysis was used to identify the predictive value of the different radiological parameters in determining pituitary adenoma consistency. The authors included 32 (84.21%) nonfunctional and 6 (15.79%) functional adenomas. The mean maximum tumor diameter was 3.7 cm, and the mean preoperative tumor volume was 16.4 cm3. Cavernous sinus invasion was present in 20 patients (52.63%). A gross-total resection was possible in 9 (23.68%) patients. The entire cohort's mean absolute tumor stiffness value was 1.8 kPa (range 1.1-3.7 kPa), whereas the mean tumor stiffness ratio was 0.66 (range 0.37-1.6). Intraoperative tumor consistency was significantly correlated with absolute and relative tumor stiffness (p = 0.0087 and 0.007, respectively). Tumor consistency alone was not a significant factor for predicting gross-total resection. Patients with intermediate and firm tumors had more complications compared to patients with soft tumors (50.00% vs 12.50%, p = 0.02) and also had longer operative times (p = 0.0002). Whereas other MRI sequences have proven to be unreliable in determining tumor consistency, MRE has been shown to be a reliable tool for predicting adenoma consistency. Preoperative knowledge of tumor consistency could be potentially useful for surgical planning, counseling about potential surgical risks, and estimating the length of operative time.


A new promising phylogenetic marker to study the diversity of fungal communities: The Glycoside Hydrolase 63 gene.
In molecular ecology, the development of efficient molecular markers for fungi remains an important research domain. Nuclear ribosomal internal transcribed spacer (ITS) region was proposed as universal DNA barcode marker for fungi, but this marker was criticized for Indel-induced alignment problems and its potential lack of phylogenetic resolution. Our main aim was to develop a new phylogenetic gene and a putative functional marker, from single-copy gene, to describe fungal diversity. Thus, we developed a series of primers to amplify a polymorphic region of the Glycoside Hydrolase GH63 gene, encoding exo-acting α-glucosidases, in basidiomycetes. These primers were validated on 125 different fungal genomic DNAs, and GH63 amplification yield was compared with that of already published functional markers targeting genes coding for laccases, N-acetylhexosaminidases, cellobiohydrolases and class II peroxidases. Specific amplicons were recovered for 95% of the fungal species tested, and GH63 amplification success was strikingly higher than rates obtained with other functional genes. We downloaded the GH63 sequences from 483 fungal genomes publicly available at the JGI mycocosm database. GH63 was present in 461 fungal genomes belonging to all phyla, except Microsporidia and Neocallimastigomycota divisions. Moreover, the phylogenetic trees built with both GH63 and Rpb1 protein sequences revealed that GH63 is also a promising phylogenetic marker. Finally, a very high proportion of GH63 proteins was predicted to be secreted. This molecular tool could be a new phylogenetic marker of fungal species as well as potential indicator of functional diversity of basidiomycetes fungal communities in term of secretory capacities.


Insight into the Lifestyle of Amoeba Willaertia magna during Bioreactor Growth Using Transcriptomics and Proteomics.
Willaertia magna C2c maky is a thermophilic free-living amoeba strain that showed ability to eliminate Legionella pneumophila, a pathogenic bacterium living in the aquatic environment. The amoeba industry has proposed the use of Willaertia magna as a natural biocide to control L. pneumophila proliferation in cooling towers. Here, transcriptomic and proteomic studies were carried out in order to expand knowledge on W. magna produced in a bioreactor. Illumina RNA-seq generated 217 million raw reads. A total of 8,790 transcripts were identified, of which 6,179 and 5,341 were assigned a function through comparisons with National Center of Biotechnology Information (NCBI) reference sequence and the Clusters of Orthologous Groups of proteins (COG) databases, respectively. To corroborate these transcriptomic data, we analyzed the W. magna proteome using LC-MS/MS. A total of 3,561 proteins were identified. The results of transcriptome and proteome analyses were highly congruent. Metabolism study showed that W. magna preferentially consumed carbohydrates and fatty acids to grow. Finally, an in-depth analysis has shown that W. magna produces several enzymes that are probably involved in the metabolism of secondary metabolites. Overall, our multi-omic study of W. magna opens the way to a better understanding of the genetics and biology of this amoeba.


Gαi2 signaling promotes skeletal muscle hypertrophy, myoblast differentiation, and muscle regeneration.
Skeletal muscle atrophy results in loss of strength and an increased risk of mortality. We found that lysophosphatidic acid, which activates a G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor, stimulated skeletal muscle hypertrophy through activation of Gα(i2). Expression of a constitutively active mutant of Gα(i2) stimulated myotube growth and differentiation, effects that required the transcription factor NFAT (nuclear factor of activated T cells) and protein kinase C. In addition, expression of the constitutively active Gα(i2) mutant inhibited atrophy caused by the cachectic cytokine TNFα (tumor necrosis factor-α) by blocking an increase in the abundance of the mRNA encoding the E3 ubiquitin ligase MuRF1 (muscle ring finger 1). Gα(i2) activation also enhanced muscle regeneration and caused a switch to oxidative fibers. Our study thus identifies a pathway that promotes skeletal muscle hypertrophy and differentiation and demonstrates that Gα(i2)-induced signaling can act as a counterbalance to MuRF1-mediated atrophy, indicating that receptors that act through Gα(i2) might represent potential targets for preventing skeletal muscle wasting.


Adherence to malaria management guidelines by health care workers in the Busoga sub-region, eastern Uganda.
Appropriate malaria management is a key malaria control strategy. The objective of this study was to determine health care worker adherence levels to malaria case management guidelines in the Busoga sub-region, Uganda. Health facility assessments, health care worker (HCW), and patient exit interview (PEI) surveys were conducted at government and private health facilities in the sub-region. All health centres (HC) IVs, IIIs, and a sample of HC IIs, representative of the tiered structure of outpatient service delivery at the district level were targeted. HCWs at these facilities were eligible for participation in the study. For PEIs, 210 patients of all ages presenting with a history of fever for outpatient care at selected facilities in each district were targeted. Patient outcome measures included testing rates, adherence to treatment, dispensing and counselling services as per national guidelines. The primary outcome was appropriate malaria case management, defined as the proportion of patients tested and only prescribed artemether-lumefantrine (AL) if positive. HCW readiness (e.g., training, supervision) and health facility capacity (e.g. availability of diagnostics and anti-malarials) to provide malaria case management were also assessed. Data were weighted to cater for the disproportionate representation of HC IIs in the study sample. A total of 3936 patients and 1718 HCW from 392 facilities were considered in the analysis. The median age of patients was 14 years; majority (63.4%) females. Most (70.1%) facilities were HCIIs and 72.7% were owned by the government. Malaria testing services were available at > 85% of facilities. AL was in stock at 300 (76.5%) facilities. Of those with a positive result, nearly all were prescribed an anti-malarial, with AL (95.1%) accounting for most prescriptions. Among those prescribed AL, 81.0% were given AL at the facility, lowest at HC IV (60.0%) and government owned (80.1%) facilities, corresponding to AL stock levels. Overall, 86.9% (95%CI 79.7, 90.7) of all enrolled patients received appropriate malaria case management. However, only 50.7% (21.2, 79.7) of patients seen at PFPs received appropriate malaria management. Adherence levels to malaria case management guidelines were good, but with gaps noted mainly in the private sector. The supply chain for AL needs to be strengthened. Interventions to improve practise at PFP facilities should be intensified.


Personalization in the health care system: do personal health budgets have an impact on outcomes and cost?
In England's National Health Service, personal health budgets are part of a growing trend to give patients more choice and control over how health care services are managed and delivered. The personal health budget programme was launched by the Department of Health in 2009, and a three-year independent evaluation was commissioned with the aim of identifying whether the initiative ensured better health- and care-related outcomes and at what cost when compared to conventional service delivery. The evaluation used a pragmatic controlled trial design to compare the outcomes and costs of patients selected to receive a personal health budget with those continuing with conventional support arrangements (control group). Just over 1000 individuals were recruited into the personal health budget group and 1000 into the control group in order to ensure sufficient statistical power, and followed for 12 months. The use of personal health budgets was associated with significant improvement in patients' care-related quality of life and psychological wellbeing at 12 months. Personal health budgets did not appear to have an impact on health status, mortality rates, health-related quality of life or costs over the same period. With net benefits measured in terms of care-related quality of life on the adult social care outcome toolkit measure, personal health budgets were cost-effective: that is, budget holders experienced greater benefits than people receiving conventional services, and the budgets were worth the cost. The evaluation provides support for the planned wider roll-out of personal health budgets in the English NHS after 2014 in so far as the localities in the pilot sample are representative of the whole country.


Prevalence of mammographically dense breasts in the United States.
National legislation is under consideration that would require women with mammographically dense breasts to be informed of their breast density and encouraged to discuss supplemental breast cancer screening with their health care providers. The number of US women potentially affected by this legislation is unknown. We determined the mammographic breast density distribution by age and body mass index (BMI) using data from 1518 599 mammograms conducted from 2007 through 2010 at mammography facilities in the Breast Cancer Surveillance Consortium (BCSC). We applied these breast density distributions to age- and BMI-specific counts of the US female population derived from the 2010 US Census and the National Health and Nutrition Examination Survey (NHANES) to estimate the number of US women with dense breasts. Overall, 43.3% (95% confidence interval [CI] = 43.1% to 43.4%) of women 40 to 74 years of age had heterogeneously or extremely dense breasts, and this proportion was inversely associated with age and BMI. Based on the age and BMI distribution of US women, we estimated that 27.6 million women (95% CI = 27.5 to 27.7 million) aged 40 to 74 years in the United States have heterogeneously or extremely dense breasts. Women aged 40 to 49 years (N = 12.3 million) accounted for 44.3% of this group. The prevalence of dense breasts among US women of common breast cancer screening ages exceeds 25 million. Policymakers and healthcare providers should consider this large prevalence when debating breast density notification legislation and designing strategies to ensure that women who are notified have opportunities to evaluate breast cancer risk and discuss and pursue supplemental screening options if deemed appropriate.


Allee effects and the spatial dynamics of a locally endangered butterfly, the high brown fritillary (Argynnis adippe).
Conservation of endangered species necessitates a full appreciation of the ecological processes affecting the regulation, limitation, and persistence of populations. These processes are influenced by birth, death, and dispersal events, and characterizing them requires careful accounting of both the deterministic and stochastic processes operating at both local and regional population levels. We combined ecological theory and observations on Allee effects by linking mathematical analysis and the spatial and temporal population dynamics patterns of a highly endangered butterfly, the high brown fritillary, Argynnis adippe. Our theoretical analysis showed that the role of density-dependent feedbacks in the presence of local immigration can influence the strength of Allee effects. Linking this theory to the analysis of the population data revealed strong evidence for both negative density dependence and Allee effects at the landscape or regional scale. These regional dynamics are predicted to be highly influenced by immigration. Using a Bayesian state-space approach, we characterized the local-scale births, deaths, and dispersal effects together with measurement and process uncertainty in the metapopulation. Some form of an Allee effect influenced almost three-quarters of these local populations. Our joint analysis of the deterministic and stochastic dynamics suggests that a conservation priority for this species would be to increase resource availability in currently occupied and, more importantly, in unoccupied sites.


Natural history of cervical neoplasia and risk of invasive cancer in women with cervical intraepithelial neoplasia 3: a retrospective cohort study.
The invasive potential of cervical intraepithelial neoplasia 3 (CIN3; also termed stage 0 carcinoma) has been poorly defined. At the National Women's Hospital, Auckland, New Zealand, treatment of CIN3 was withheld from a substantial number of women between 1965 and 1974 as part of an unethical clinical study. The resulting variation in management allows comparison of the long-term risk of invasive cancer of the cervix in women whose lesion was minimally disturbed with those who had adequate initial treatment followed by conventional management. We aimed to estimate the long-term risk of invasive cancer in these two groups of women. A judicial inquiry referred for independent clinical review in 1988 all women for whom there remained doubt about the adequacy of their management. Between February, 2001, and December, 2004, medical records, cytology, and histopathology were reviewed for all women with CIN3 diagnosed between 1955 and 1976, whose treatment was reviewed by judicial inquiry and whose medical records could be located, and linkages were done with cancer and death registers and electoral rolls. To take into account the probability that the CIN3 lesion had been completely removed, we classified adequacy of treatment by type of procedure, presence of CIN3 at the excision margin, and subsequent cytology. The primary outcome was cumulative incidence of invasive cancer of the cervix or vaginal vault. Follow-up continued until death or Dec 31, 2000, whichever came first. Analyses accounted for procedures during follow-up. 1229 women whose treatment was reviewed by the judicial inquiry in 1987-88 were included. Of these, 48 records (4%) could not be located and 47 women (4%) did not meet the inclusion criteria. At histopathological review, a further 71 (6% of 1134) women were excluded because the review diagnosis was not CIN3. We identified outcomes in the remaining 1063 (86% of 1229) women diagnosed with CIN3 at the hospital in 1955-76. In 143 women managed only by punch or wedge biopsy, cumulative incidence of invasive cancer of the cervix or vaginal vault was 31.3% (95% CI 22.7-42.3) at 30 years, and 50.3% (37.3-64.9) in the subset of 92 such women who had persistent disease within 24 months. However, cancer risk at 30 years was only 0.7% (0.3-1.9) in 593 women whose initial treatment was deemed adequate or probably adequate, and whose treatment for recurrent disease was conventional. This study provides the most valid direct estimates yet available of the rate of progression from CIN3 to invasive cancer. Women with untreated CIN3 are at high risk of cervical cancer, whereas the risk is very low in women treated conventionally throughout.


The Nissl substance of living and fixed spinal ganglion cells. I. A phase contrast study.
Living chick embryo spinal ganglion neurons grown from 1 to 4 weeks in vitro were studied under the phase contrast microscope. In the peripheral cytoplasm of the earliest stages studied, a homogeneous, phase-dense material is seen which corresponds in location to the cytoplasmic basophil material of the same stages. As maturation proceeds, this material increases in extent, and becomes separated by lighter channels into discrete bodies. Short fixation by 1 per cent buffered osmium tetroxide followed by post-fixation with neutral buffered formalin does not significantly alter the size, shape, or distribution of any of the cytoplasmic components, and the fixed, hydrated cell is almost indistinguishable from the living cell. Dehydration causes some shrinkage of the fixed preparations, but if the photographs of the stained preparations are enlarged to correspond with those of the living cell, excellent correspondence can be made between at least the larger basophil masses and the larger dark masses seen with phase contrast. Fixation by a formalin-mercuric chloride procedure also results in satisfactory correspondence between the stained Nissl bodies and the phase-dark homogeneous areas. It is concluded that discrete Nissl bodies preexist in the living neuron and are essentially unchanged after good cytological fixation. Evidence is also presented of the presence of neurofibrils in the living state.


The gene for the axonal cell adhesion molecule TAX-1 is amplified and aberrantly expressed in malignant gliomas.
The human TAX-1 gene encodes a Mr 135,000 glycoprotein that is transiently expressed on the surface of a subset of neurons during development and is involved in neurite outgrowth. The TAX-1 gene has been mapped to a region on chromosome 1 that has been implicated in microcephaly and the Van der Woude syndrome. Using restriction landmark genome scanning to search for amplified genes in gliomas, we found TAX-1 to be amplified in 2 high-grade gliomas among a group of 26 gliomas investigated. Real-time reverse transcription-quantitative PCR analysis detected high levels of TAX-1 mRNA in glial tumors, even in the absence of TAX-1 gene amplification. Immunohistochemical analysis revealed abundant levels of TAX-1 in neoplastic glial cells of glioblastoma multiforme tumors. Because glial tumors are highly invasive and in view of the role of TAX-1 in neurite outgrowth, we investigated the potential role of TAX-1 in glioma cell migration. Using an in vitro assay, we found that the migration of glioma tumor cells is profoundly reduced in the presence of either an anti-TAX-1 antibody or a TAX-1 antisense oligonucleotide. Our findings suggest that TAX-1 plays a role in glial tumorigenesis and may provide a potential target for therapeutic intervention.


The association between exposure to a rear-end collision and future neck or shoulder pain: a cohort study.
Neck pain is the most frequently reported feature in connection with whiplash injury, but it is also a common complaint in the general population. Therefore it is crucial to include an unexposed comparison group when evaluating the association between neck pain and a previous motor vehicle crash (MVC). To determine whether exposure to a rear-end collision, without or with whiplash injury, is associated with future neck or shoulder pain, a cohort study was conducted. The study population consisted of persons covered by traffic insurance at one of the largest insurance companies in Sweden. Claim reports were collected from the period November 1987 to April 1988. Drivers exposed to a rear-end collision were divided into two subgroups, without reported whiplash injury (n = 204) and with reported whiplash injury (n = 232). Two comparison groups, unexposed to MVCs, consisting of 1599 and 2089 persons, were selected with consideration taken to the age and gender distribution in the exposed subgroups. A questionnaire concerning neck or shoulder pain and other subjective health complaints was mailed to all the study subjects at follow-up in 1994, 7 years after the rear-end collision. The relative risk of neck or shoulder pain at follow-up was 1.3 (95% CI 0.8-2.0) in the exposed subjects without whiplash injury compared with the unexposed. The corresponding relative risk in subjects with whiplash injury was 2.7 (95% CI 2.1-3. 5). We conclude that there is no increased risk of future neck or shoulder pain in drivers who did not report whiplash injury in connection with a rear-end collision 7 years earlier. In drivers with reported whiplash injury, the risk of neck or shoulder pain 7 years after the collision was increased nearly three-fold compared with that in unexposed subjects.


Effects of environmental impact labels on the sustainability of food purchases: Two randomised controlled trials in an experimental online supermarket.
Providing consumers with product-specific environmental impact information for food products (ecolabels) may promote more sustainable purchasing, needed to meet global environmental targets. Two UK studies investigated the effectiveness of different ecolabels using an experimental online supermarket platform. Study 1 (N = 1051 participants) compared three labels against control (no label), while Study 2 (N = 4979) tested four designs against control. Study 1 found significant reductions in the environmental impact score (EIS) for all labels compared to control (labels presented: values for four environmental indicators [-3.9 percentiles, 95%CIs: -5.2,-2.6]; a composite score [taking values from A to E; -3.9, 95%CIs: -5.2,-2.5]; or both together [-3.2, 95%CIs: -4.5,-1.9]). Study 2 showed significant reductions in EIS compared to control for A-E labels [-2.3, 95%CIs: -3.0,-1.5], coloured globes with A-E scores [-3.2, 95%CIs:-3.9,-2.4], and red globes highlighting 'worse' products [-3.2, 95%CIs:-3.9,-2.5]. There was no evidence that green globes highlighting 'better' products were effective [-0.5, 95%CIs:-1.3,0.2]. Providing ecolabels is a promising intervention to promote the selection of more sustainable products.


AceView: a comprehensive cDNA-supported gene and transcripts annotation.
Regions covering one percent of the genome, selected by ENCODE for extensive analysis, were annotated by the HAVANA/Gencode group with high quality transcripts, thus defining a benchmark. The ENCODE Genome Annotation Assessment Project (EGASP) competition aimed at reproducing Gencode and finding new genes. The organizers evaluated the protein predictions in depth. We present a complementary analysis of the mRNAs, including alternative transcript variants. We evaluate 25 gene tracks from the University of California Santa Cruz (UCSC) genome browser. We either distinguish or collapse the alternative splice variants, and compare the genomic coordinates of exons, introns and nucleotides. Whole mRNA models, seen as chains of introns, are sorted to find the best matching pairs, and compared so that each mRNA is used only once. At the mRNA level, AceView is by far the closest to Gencode: the vast majority of transcripts of the two methods, including alternative variants, are identical. At the protein level, however, due to a lack of experimental data, our predictions differ: Gencode annotates proteins in only 41% of the mRNAs whereas AceView does so in virtually all. We describe the driving principles of AceView, and how, by performing hand-supervised automatic annotation, we solve the combinatorial splicing problem and summarize all of GenBank, dbEST and RefSeq into a genome-wide non-redundant but comprehensive cDNA-supported transcriptome. AceView accuracy is now validated by Gencode. Relative to a consensus mRNA catalog constructed from all evidence-based annotations, Gencode and AceView have 81% and 84% sensitivity, and 74% and 73% specificity, respectively. This close agreement validates a richer view of the human transcriptome, with three to five times more transcripts than in UCSC Known Genes (sensitivity 28%), RefSeq (sensitivity 21%) or Ensembl (sensitivity 19%).


Bacterial Canker Caused by Clavibacter michiganensis subsp. michiganensis on Greenhouse-Grown Tomato in the Western Mediterranean Region of Turkey.
During the 2003 winter growing season, a disease occurred on tomato (Lycopersicon esculentum L.) cvs. Selin, Astona, 198 RN, Malike, and Tulin that were growing in almost all greenhouses of Serik, Aksu, Kumluca, Demre, and Kinik in the Antalya Province, and Keçiborlu, Çandir, and Şeyhler in the Isparta Province in the western Mediterranean Region of Turkey. Disease incidence ranged from 26 to 65%, resulting in economically heavy losses. Symptoms were water-soaked, dark brown-to-black lesions on the leaf margins and asymmetrical wilting of the leaflets. In advanced stages of disease, vascular tissue had a light brown discoloration. A gram-positive bacterium was consistently isolated and formed light gray colonies with internal light gray flecks approximately 2 to 3 mm in diameter on the modified semiselective medium (SCM) (2). Thirty isolates were identified as Clavibacter michiganensis subsp. michiganensis using polymerase chain reaction (PCR) with primers CMM 5 and CMM 6 on the basis of consistent amplification of a 614-bp DNA fragment (1). The isolates were also identified as C. michiganensis subsp. michiganensis (similarity 0.810 to 0.888%) using fatty acid methyl ester (FAME) analysis and Sherlock Microbial Identification System software (Microbial ID, Newark, DE). A pathogenicity test was done by spraying a 6-week-old tomato seedling (cv. Selin) with the bacterial concentration of 108 CFU/ml in sterile water. Control tomato seedlings were only sprayed with sterile water. The inoculated plants were covered with polyethylene bags for 48 h and kept in a growth chamber at 25°C. Symptoms developed on the inoculated plants within approximately 10 days and were similar to those observed in the greenhouse. No symptoms developed on control plants. The bacterium was reisolated from inoculated plants, and its identity was confirmed by colony type on SCM, PCR, and FAME analysis. Although it has been known to be present previously in Aegean, eastern Mediterranean, and the eastern Anatolia regions of Turkey, to our knowledge, this is the first report of bacterial canker on greenhouse tomatoes in the western Mediterranean Region of Turkey. Occurrence of the disease in almost all greenhouses may be the result of the high relative humidity present and the lack of protective bactericide applications in many greenhouses of the region. References: (1) J. Dreier et al. Phytopathology 85:462, 1995. (2) M. Fatmi and N. W. Schaad. Phytopathology 78:121, 1988.


LRF-mediated Dll4 repression in erythroblasts is necessary for hematopoietic stem cell maintenance.
Hematopoietic stem cells (HSCs) are the most primitive cells in the hematopoietic system and are under tight regulation for self-renewal and differentiation. Notch signals are essential for the emergence of definitive hematopoiesis in mouse embryos and are critical regulators of lymphoid lineage fate determination. However, it remains unclear how Notch regulates the balance between HSC self-renewal and differentiation in the adult bone marrow (BM). Here we report a novel mechanism that prevents HSCs from undergoing premature lymphoid differentiation in BM. Using a series of in vivo mouse models and functional HSC assays, we show that leukemia/lymphoma related factor (LRF) is necessary for HSC maintenance by functioning as an erythroid-specific repressor of Delta-like 4 (Dll4) expression. Lrf deletion in erythroblasts promoted up-regulation of Dll4 in erythroblasts, sensitizing HSCs to T-cell instructive signals in the BM. Our study reveals novel cross-talk between HSCs and erythroblasts, and sheds a new light on the regulatory mechanisms regulating the balance between HSC self-renewal and differentiation.


Associations Between Materialism, Gratitude, and Well-Being in Children of Overseas Filipino Workers.
Children left behind by parents who are overseas Filipino workers (OFW) benefit from parental migration because their financial status improves. However, OFW families might emphasize the economic benefits to compensate for their separation, which might lead to materialism among children left behind. Previous research indicates that materialism is associated with lower well-being. The theory is that materialism focuses attention on comparing one's possessions to others, making one constantly dissatisfied and wanting more. Research also suggests that gratitude mediates this link, with the focus on acquiring more possessions that make one less grateful for current possessions. This study explores the links between materialism, gratitude, and well-being among 129 adolescent children of OFWs. The participants completed measures of materialism, gratitude, and well-being (life satisfaction, self-esteem, positive and negative affect). Results showed that gratitude mediated the negative relationship between materialism and well-being (and its positive relationship with negative affect). Children of OFWs who have strong materialist orientation seek well-being from possessions they do not have and might find it difficult to be grateful of their situation, contributing to lower well-being. The findings provide further evidence for the mediated relationship between materialism and well-being in a population that has not been previously studied in the related literature. The findings also point to two possible targets for psychosocial interventions for families and children of OFWs.


Personal accounts of exercise and quality of life from the perspective of breast cancer survivors.
The purpose of the study was to examine breast cancer survivors' perceptions of exercise and their quality of life (QoL). About 289 breast cancer survivors completed a survey addressing exercise attitudes, behaviour and perceived QoL. In addition, the breast cancer survivors completed two open-ended questions designed to explore perceptions of exercise and QoL throughout their cancer experience. Inductive and deductive content analyses were used to analyse responses. About 19 first-order themes were identified, which were clustered into five second-order themes that included; Exercise behaviour, Lifestyle, Limitations and barriers, Growth and priorities, and Personal beliefs and values. The findings identified a framework of multifaceted views held by breast cancer survivors in relation to their QoL and outlook on their disease. Given that cancer survivors are faced with a number of treatment related morbidities 2 years post-diagnosis, there is a need for health professionals to carefully address a cancer survivor's exercise needs in an attempt to help improve their future QoL.


On the effect of response transformations in sequential parameter optimization.
Parameter tuning of evolutionary algorithms (EAs) is attracting more and more interest. In particular, the sequential parameter optimization (SPO) framework for the model-assisted tuning of stochastic optimizers has resulted in established parameter tuning algorithms. In this paper, we enhance the SPO framework by introducing transformation steps before the response aggregation and before the actual modeling. Based on design-of-experiments techniques, we empirically analyze the effect of integrating different transformations. We show that in particular, a rank transformation of the responses provides significant improvements. A deeper analysis of the resulting models and additional experiments with adaptive procedures indicates that the rank and the Box-Cox transformation are able to improve the properties of the resultant distributions with respect to symmetry and normality of the residuals. Moreover, model-based effect plots document a higher discriminatory power obtained by the rank transformation.


Preference‒performance linkage in the diamondback moth, Plutella xylostella, and implications for its management.
Host plants affect development, survival, and reproduction of phytophagous insects. In the case of holometabolous species, whose larvae have little mobility to find a host plant, the ability of females to discriminate hosts on the basis of their nutritional quality may be an important factor determining insect performance. The preference‒performance correlation hypothesis states that females will choose to lay their eggs on host plants that provide the best offspring performance. The effects of three cultivated and two wild brassicas (Brassicales: Brassicaceae) on the biology of the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), an important pest of brassicas, were investigated. Based on these data, the preference-performance correlation hypothesis was tested. The results allowed the discussion of the possible role of wild brassicas on population dynamics of the pest. The life table parameters net reproduction rate and intrinsic rate of increase were used as indicatives of insect performance because they provide a detailed description of the survivorship, development, and reproduction of a population. Development, survival, and reproduction were affected by the cultivated and wild brassicas. Both net reproduction rate and intrinsic rate of increase were lower in individuals fed on wild brassicas, which indicates that brassicas are not nutritionally suitable for P. xylostella. Nevertheless, females showed no oviposition preference among host plants. The results showed that host plant quality might not be the only factor determining host selection by female P. xylostella. Results also suggest that wild brassicas may serve as a refuge for P. xylostella, favoring pest survival when crops are disturbed by insecticide application, irrigation, or ploughing.


A checkpoint cliffhanger at the dawn of placental mammals.
The PD-1 ligands PD-L1 and PD-L2 are commonly expressed on the surface of cells, where they regulate immune system activation. However, the specific role played by each ligand has been unclear. Using site-directed mutagenesis, surface plasmon resonance, and crystallography, Philips et al. explore the distinct features of PD-L2 and identify a specific evolutionary event linked to its appearance. This work provides a deeper understanding of how the immune system adapted to mammalian placental gestation and could be an important consideration in the development of new immune checkpoint therapies.


Variants in Miro1 Cause Alterations of ER-Mitochondria Contact Sites in Fibroblasts from Parkinson's Disease Patients.
Although most cases of Parkinson´s disease (PD) are idiopathic with unknown cause, an increasing number of genes and genetic risk factors have been discovered that play a role in PD pathogenesis. Many of the PD-associated proteins are involved in mitochondrial quality control, e.g., PINK1, Parkin, and LRRK2, which were recently identified as regulators of mitochondrial-endoplasmic reticulum (ER) contact sites (MERCs) linking mitochondrial homeostasis to intracellular calcium handling. In this context, Miro1 is increasingly recognized to play a role in PD pathology. Recently, we identified the first PD patients carrying mutations in RHOT1, the gene coding for Miro1. Here, we describe two novel RHOT1 mutations identified in two PD patients and the characterization of the cellular phenotypes. Using whole exome sequencing we identified two PD patients carrying heterozygous mutations leading to the amino acid exchanges T351A and T610A in Miro1. We analyzed calcium homeostasis and MERCs in detail by live cell imaging and immunocytochemistry in patient-derived fibroblasts. We show that fibroblasts expressing mutant T351A or T610A Miro1 display impaired calcium homeostasis and a reduced amount of MERCs. All fibroblast lines from patients with pathogenic variants in Miro1, revealed alterations of the structure of MERCs. Our data suggest that Miro1 is important for the regulation of the structure and function of MERCs. Moreover, our study supports the role of MERCs in the pathogenesis of PD and further establishes variants in RHOT1 as rare genetic risk factors for neurodegeneration.


The role of prefrontal cortex in object-in-place learning in monkeys.
Previous ablation studies in monkeys suggest that prefrontal cortex is involved in a wide range of learning and memory tasks. However, monkeys with crossed unilateral lesions of frontal and temporal cortex are unimpaired at concurrent object-reward association learning but are impaired at conditional learning and the implementation of memory-based performance rules. We trained seven monkeys preoperatively on an associative learning task that required them to associate objects embedded in unique complex scenes with reward. Three monkeys then had crossed unilateral lesions of frontal and inferior temporal cortex and the remaining monkeys had bilateral prefrontal cortex ablation. Both groups were severely impaired postoperatively. These results show that both bilateral prefrontal cortex ablation and frontal-temporal disconnection impair associative learning for objects embedded in scenes. The results provide evidence that the function of frontal-temporal interactions in memory is not limited to conditional learning tasks and memory-dependent performance rules. We propose that rapid object-in-place learning requires the interaction of frontal cortex with inferotemporal cortex because visual object and contextual information which is captured over multiple saccades must be processed as a unique complex event that is extended in time. The present results suggest a role for frontal-temporal interaction in the integration of visual information over time.


Acoustic injury and TRPV1 expression in the cochlear spiral ganglion.
Acoustic trauma not only produces temporary and permanent hearing loss but is a common cause of chronic tinnitus. Recent work indicated a possible role for the transient receptor potential channel vanilloid subfamily type 1 (TRPV1) in modulating the effects of cochlear injury. In our research, we investigated the effects of acoustic damage on TRPV1 expression in spiral ganglion neurons of adult rats. After exposing them unilaterally to noise, we extracted cochleas and processed the spiral ganglion for TRPV1 expression at four posttrauma intervals (2 hours, 24 hours, 12 days, and 16.9 months). We measured TRPV1 immunodensity in the apical, middle, and basal turns of the cochlea. We found a significant interaction (p = .039) between posttrauma interval and regional cochlear receptor expression: For survival intervals between 24 hours and 2 weeks, TRPV1 density increased in all cochlear regions; at the longest survival interval (16.9 months), TRPV1 density was dramatically reduced in the basal region. We also psychophysically tested the long-survival subjects, which showed evidence of 20-kHz tonal tinnitus. These results suggest that TRPV1 may participate after cochlear injury in a signal cascade that is responsible for the neuroplastic events leading to tinnitus and hyperacusis.


Targeted delivery of the hydroxylase inhibitor DMOG provides enhanced efficacy with reduced systemic exposure in a murine model of colitis.
Targeting hypoxia-sensitive pathways has recently been proposed as a new therapeutic approach to the treatment of intestinal inflammation. HIF-hydroxylases are enzymes which confer hypoxic-sensitivity upon the hypoxia-inducible factor (HIF), a major regulator of the adaptive response to hypoxia. Previous studies have shown that systemic (intraperitoneal) administration of hydroxylase inhibitors such as dimethyloxalylglycine (DMOG) is profoundly protective in multiple models of colitis, however the therapeutic potential of this approach is limited due to potential side-effects associated with systemic drug exposure and the fact that orally delivered DMOG is ineffective (likely due to drug inactivation by gastric acid). In order to overcome these issues, we formulated DMOG in a liquid emulsion drug delivery system which, when coated with specific polymer coatings, permits oral delivery of a reduced dose which is released locally throughout the colon. This colon-targeted DMOG formulation demonstrated increased relative colonic bioactivity with reduced systemic exposure and provided a similar degree of protection to systemic (intraperitoneal) administration at a 40-fold lower dose in DSS-induced colitis. In summary, targeted delivery of DMOG to the colon provides local protection resulting in enhanced efficacy with reduced systemic exposure in the treatment of colitis. This novel approach to targeting hydroxylase inhibitors to specific diseased regions of the GI tract may improve it's potential as a new therapeutic in inflammatory bowel diseases such as ulcerative colitis.


Misperception of tiredness in young adults with insomnia.
People with insomnia often exhibit interpretive biases to cues associated with their condition. This study examined whether individuals with insomnia display an interpretive bias, such that they misperceive facial attributes of tiredness in a disorder-consistent manner. The efficacy of providing feedback related to the accuracy of participants' perception on later judgements of tiredness was further examined. Forty participants, 20 with DSM-5-defined insomnia disorder and 20 normal-sleepers, participated. The perception of one's own facial appearance of tiredness was assessed twice over two consecutive days using a visual task whereby participants indicated when a morphing image of their face represented their current level of tiredness. Visual and verbal feedback, related to participants' degree of misperception, was provided on completion of Day 1 testing. Overall, individuals with insomnia perceived their own face as significantly more tired than a baseline neutral photograph was, whereas normal-sleepers perceived themselves as appearing more alert. This pattern of results was only apparent on Day 1. Although no group × day interaction was established, mean scores suggest an improvement in perception on Day 2 amongst individuals with insomnia only. These findings suggest that individuals with insomnia exhibit a misperception of their facial attributes of tiredness, interpreting them in a disorder-consistent manner. This finding adds to the body of literature on cognitive models of insomnia, demonstrating more general cognitive biases in the disorder. Further, the results provide suggestive evidence that this misperception may be reformed.


Genetic monitoring of western corn rootworm (Coleoptera: Chrysomelidae) populations on a microgeographic scale.
Microsatellite and mitochondrial DNA genetic monitoring of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was undertaken in Croatia and Serbia from 1996 to 2011 and in the United States in 2011. The seven U.S. populations displayed the greatest allelic diversity. In Europe, the highest number of alleles was found in Rugvica, Croatia, and Surčin, Serbia, the two sites closest to international airports. The highest number of mitochondrial (mt) DNA haplotypes was recorded from Croatia in 1996. From 2009 to 2011, haplotype diversity declined, and Croatia and Serbia had a single fixed haplotype. U.S. continuous maize locations had one haplotype, while three haplotypes were found at crop-rotated locations. Minimal temporal genetic differentiation was found within and between populations in Europe and the United States. Bayesian cluster analysis identified two genetic clusters that grouped western corn rootworm from Croatia and Serbia separately from U.S. populations; however, these clusters were not neat, and numerous U.S. individuals had both European and U.S. ancestry, suggesting bidirectional gene flow. Bottlenecks were identified within most Croatian populations sampled in 1996, only two populations in 2009, and in all populations in 2011. Bottlenecks were not identified from Serbia from 1996 to 2011 or from the United States in 2011. As suspected Serbia was identified as the geographic source of western corn rootworm in Croatia. The temporal genetic monitoring undertaken allowed a deeper understanding of the population genetics of western corn rootworm in Croatia, neighboring Serbia, and its geographic source in the United States. The data obtained can be used to inform western corn rootworm pest management strategies in Croatia and Europe.


Dysfunctional peripheral T follicular helper cells dominate in people with impaired influenza vaccine responses: Results from the FLORAH study.
Antigen-primed cluster of differentiation (CD) 4+ T follicular helper (Tfh) cells interact with B cells in the germinal centers (GCs) of lymph nodes to generate vaccine-induced antibody (Ab) responses. In the circulation, peripheral Tfh (pTfh) cells, a subset of memory CD4 T cells, serve as surrogates for GC Tfh because of several functional and phenotypic similarities between them. We investigated features of H1N1 influenza antigen-specific pTfh (Ag.pTfh) in virologically controlled HIV+ volunteers on antiretroviral therapy (ART) and healthy control (HC) participants selected from a seasonal influenza vaccine responsiveness study. Selection of the participants was made based on age, defined as young (18-40 y) and old (>60 y) and on their classification as a vaccine responder (VR) or vaccine nonresponder (VNR). VRs demonstrated expansion of CD40L+ and CD69+ Ag.pTfh, with induction of intracellular interleukin 21 (IL-21) and inducible costimulator (ICOS) post vaccination; these responses were strongest in young HC VRs and were less prominent in HIV+ individuals of all ages. Ag.pTfh in VNRs exhibited dramatically different characteristics from VRs, displaying an altered phenotype and a cytokine profile dominated by cytokines IL-2, tumor necrosis factor alpha (TNF-α), or IL-17 but lacking in IL-21. In coculture experiments, sorted pTfh did not support the B cell IgG production in VNRs and were predominantly an inflammatory T helper 1 (Th1)/T helper 17 (Th17) phenotype with lower ICOS and higher programmed cell death protein 1 (PD1) expression. Induction of IL-21 and ICOS on Ag.pTfh cells are negatively affected by both aging and HIV infection. Our findings demonstrate that dysfunctional Ag.pTfh cells with an altered IL-21/IL-2 axis contribute to inadequate vaccine responses. Approaches for targeting inflammation or expanding functional Tfh may improve vaccine responses in healthy aging and those aging with HIV infection.


A history of the DSM-5 Scientific Review Committee.
This article describes the history of the Scientific Review Committee (SRC) for DSM-5 and reviews its background, procedures and deliberative processes, and conceptual/philosophical framework. The results of its work and the most important and contentious issues that arose in its efforts are reviewed. The central role of the SRC was to provide external review for all proposals for diagnostic change in DSM-5, evaluate them on their level of empirical support using objectively structured rules of evidence agreed upon in advance and make appropriate recommendations to the leadership of the American Psychiatric Association. While the creation of the SRC necessitated a great deal of additional work on the part of the SRC, the workgroups and the DSM-5 Task Force, the SRC succeeded in increasing the focus on empirical standards for nosologic change and providing a greater degree of consistency and objectivity in the DSM review process. The article concludes with recommendations, based on lessons learned, for similar efforts that might be included in future iterations of our psychiatric nosology.


Increased myocardial matrix metalloproteinases in hypoxic newborn pigs during resuscitation: effects of oxygen and carbon dioxide.
Perinatal asphyxia is associated with cardiac dysfunction, and it is important to prevent further tissue injury during resuscitation. There is increasing evidence that myocardial matrix metalloproteinases (MMPs) are involved in myocardial hypoxaemia-reoxygenation injury. To assess MMPs and antioxidant capacity in newborn pigs after global ischaemia and subsequent resuscitation with ambient air or 100% O(2) at different PaCO(2)-levels. Newborn pigs (12-36 h of age) were resuscitated for 30 min by ventilation with 21% or 100% O(2) at different PaCO(2) levels after a hypoxic insult, and thereafter observed for 150 min. In myocardial tissue extracts, MMPs were analyzed by gelatin zymography and broad matrix-degrading capacity (total MMP). Total endogenous antioxidant capacity in myocardial tissue extracts was measured by the oxygen radical absorbance capacity (ORAC) assay. Matrix metalloproteinase-2 more than doubled from baseline values (P < 0.001), and was higher in piglets resuscitated with 100% O(2) than with ambient air (P = 0.012). The ORAC value was considerably decreased (P < 0.001). In piglets with elevated PaCO(2), total MMP-activity in the right ventricle was more increased than in the left ventricle (P = 0.008). In the left ventricle, total MMPactivity was higher in the piglets with low PaCO(2) than in the piglets with elevated PaCO(2) (P = 0.013). In hypoxaemia-reoxygenation injury the MMP-2 level was highly increased and was most elevated in the piglets resuscitated with 100% O(2). Antioxidant capacity was considerably decreased. Assessed by total MMP-activity, elevated PaCO(2) during resuscitation might protect the left ventricle, and probably increase right ventricle injury of the myocardium.


Malaria Policy Advisory Committee to the WHO: conclusions and recommendations of seventh biannual meeting (March 2015).
The Malaria Policy Advisory Committee to the World Health Organization held its seventh meeting in Geneva, Switzerland from 5 to 7 March 2015. This article provides a summary of the discussions, conclusions and meeting recommendations. Meeting sessions included: an update on the Greater Mekong Subregion elimination strategy; an update on the RTS,S vaccine; G6PD testing to support the safe use of anti-relapse therapy for Plasmodium vivax; update from the Vector Control Advisory Group; newly proposed evidence reviews or consultations on malaria terminology, malaria in pregnancy, and the feasibility of eradication; as well as updates from the World Health Organization Global Malaria Programme regarding their strategy update and policy setting processes. Policy statements, position statements, and guidelines that arise from the Malaria Policy Advisory Committee meeting conclusions and recommendations will be formally issued and disseminated to World Health Organization Member States by the World Health Organization Global Malaria Programme.


Mechanisms of Very Late Drug-Eluting Stent Thrombosis Assessed by Optical Coherence Tomography.
The pathomechanisms underlying very late stent thrombosis (VLST) after implantation of drug-eluting stents (DES) are incompletely understood. Using optical coherence tomography, we investigated potential causes of this adverse event. Between August 2010 and December 2014, 64 patients were investigated at the time point of VLST as part of an international optical coherence tomography registry. Optical coherence tomography pullbacks were performed after restoration of flow and analyzed at 0.4 mm. A total of 38 early- and 20 newer-generation drug-eluting stents were suitable for analysis. VLST occurred at a median of 4.7 years (interquartile range, 3.1-7.5 years). An underlying putative cause by optical coherence tomography was identified in 98% of cases. The most frequent findings were strut malapposition (34.5%), neoatherosclerosis (27.6%), uncovered struts (12.1%), and stent underexpansion (6.9%). Uncovered and malapposed struts were more frequent in thrombosed compared with nonthrombosed regions (ratio of percentages, 8.26; 95% confidence interval, 6.82-10.04; P<0.001 and 13.03; 95% confidence interval, 10.13-16.93; P<0.001, respectively). The maximal length of malapposed or uncovered struts (3.40 mm; 95% confidence interval, 2.55-4.25; versus 1.29 mm; 95% confidence interval, 0.81-1.77; P<0.001), but not the maximal or average axial malapposition distance, was greater in thrombosed compared with nonthrombosed segments. The associations of both uncovered and malapposed struts with thrombus were consistent among early- and newer-generation drug-eluting stents. The leading associated findings in VLST patients in descending order were malapposition, neoatherosclerosis, uncovered struts, and stent underexpansion without differences between patients treated with early- and new-generation drug-eluting stents. The longitudinal extension of malapposed and uncovered stent was the most important correlate of thrombus formation in VLST.


Assessing the impact of behavioral risk factors and known-groups validity of the SF-12 in a US Chinese immigrant population.
Health status measures are being used in increasingly diverse populations. However, there are no known studies to date that examine the SF-12 in US Chinese populations. This study reports on the performance and validity of the SF-12 among Chinese immigrants residing in New York City, and evaluates the impact of multiple behavioral risk factors on physical and mental health status. We used cross-sectional survey data from a multistage probability sample of 2537 Chinese adults. SF-12 scores were examined according to sociodemographic, cultural, and clinical characteristics. Regression analyses were used to examine associations between health status and co-occurring behavioral risk factors of smoking, risky drinking, physical inactivity, and overweight/obesity. SF-12 scores were significantly lower among women, those with less education, lower incomes, and more health problems (P < 0.001). Older adults had worse physical but better mental health (P < 0.05). Individuals with 1, 2, 3, and 4 behavioral risk factors reported decreases of 1.91, 2.92, 4.86, and 9.21 points on the PCS-12, respectively, in comparison with the reference group having zero risk factors (P < 0.01). Similar trends up to 2 co-occurring risks were observed with MCS-12 scores (P < 0.01). The SF-12 exhibited known-groups validity in a US Chinese immigrant population. Co-occurring behavioral risk factors were associated with progressive declines in physical health, independent of sociodemographic and clinical characteristics traditionally associated with impairments in health status. Targeting patients with multiple risks for behavior change may be effective in improving health across diverse populations.


Flow cytometric analysis of marine bacteria with hoechst 33342.
We investigated the accuracy and precision of flow cytometric (FCM) estimates of bacterial abundances using 4', 6-diamidino-2-phenylindole (DAPI) and Hoechst 33342 (HO342, a bisbenzamide derivative) on paraformaldehyde-fixed seawater samples collected from two stations near Oahu, Hawaii. The accuracy of FCM estimates was assessed against direct counts by using epifluorescence microscopy. DAPI and HO342 differ in two aspects of their chemistry that make HO342 better suited for staining marine heterotrophic bacteria for FCM analysis. These differences are most important in studies of open-ocean ecosystems that require dual-beam FCM analysis to clearly separate heterotrophic bacterial populations from populations of photosynthetic Prochlorococcus spp. Bacterial populations were easier to distinguish from background fluorescence when stained with HO342 than when stained with DAPI, because HO342 has a higher relative fluorescence quantum yield. A substantially higher coefficient of variation of blue fluorescence, which was probably due to fluorescent complexes formed by DAPI with double-stranded RNA, was observed for DAPI-stained populations. FCM estimates averaged 2.0 and 12% higher than corresponding epifluorescence microscopy direct counts for HO342 and DAPI-stained samples, respectively. A paired-sample t test between FCM estimates and direct counts found no significant difference for HO342-stained samples but a significant difference for DAPI-stained samples. Coefficients of variation of replicate FCM abundance estimates ranged from 0.63 to 2.9% (average, 1.5%) for natural bacterial concentrations of 6 x 10 to 15 x 10 cells ml.


A randomized controlled trial to evaluate the effectiveness and cost-effectiveness of psychodynamic counselling for general practice patients with chronic depression.
Counsellors have been employed in general practice with little evidence of effectiveness. This study examined the effectiveness and cost-effectiveness of short-term counselling in general practice for patients with chronic depression either alone or combined with anxiety. A randomized controlled trial with an economic evaluation was carried out in Derbyshire. One hundred and forty-five patients were recruited at seven GP practices by screening using the Beck Depression Inventory. Both the experimental and control group received routine GP treatment but the experimental group were also referred to the practice counsellor. Depression, anxiety and other mental health symptoms, social and interpersonal functioning and social support were measured at baseline, 6 months and 12 months. Comprehensive costs were also estimated. There was an overall significant improvement in the actual scores over time, but there were no significant differences between the two groups on any of the measures at either 6 or 12 months. However fewer experimental group patients were still 'cases' on the BDI than controls at 12 months. There were no significant differences in the mean total costs, aggregate costs of services, or any service-group costs except for primary care, between the experimental and control groups over time. This trial demonstrates only very limited evidence of improved outcomes in those referred to counselling and increased primary care treatment costs in the short-term. Stricter referral criteria to exclude the more severely depressed in the group (BDI > or = 24) might have yielded more conclusive results.


New Insight Into Metformin Action: Regulation of ChREBP and FOXO1 Activities in Endothelial Cells.
Metformin has been considered a potential adjunctive therapy in treating poorly controlled type 1 diabetes with obesity and insulin resistance, owing to its potent effects on improving insulin sensitivity. However, the underlying mechanism of metformin's vascular protective effects remains obscure. Thioredoxin-interacting protein (TXNIP), a key regulator of cellular redox state induced by high-glucose concentration, decreases thioredoxin reductase activity and mediates apoptosis induced by oxidative stress. Here we report that high glucose-induced endothelial dysfunction is associated with induction of TXNIP expression in primary human aortic endothelial cells exposed to high-glucose conditions, whereas the metformin treatment suppresses high-glucose-induced TXNIP expression at mRNA and protein levels. We further show that metformin decreases the high-glucose-stimulated nuclear entry rate of two transcription factors, carbohydrate response element-binding protein (ChREBP) and forkhead box O1 (FOXO1), as well as their recruitment on the TXNIP promoter. An AMP-activated protein kinase inhibitor partially compromised these metformin effects. Our data suggest that endothelial dysfunction resulting from high-glucose concentrations is associated with TXNIP expression. Metformin down-regulates high-glucose-induced TXNIP transcription by inactivating ChREBP and FOXO1 in endothelial cells, partially through AMP-activated protein kinase activation.


Co-release of acetylcholine and GABA by the starburst amacrine cells.
Rabbit retinas were isolated from the eye and maintained in vitro. When they were incubated for 60 min in the presence of 3H-GABA, subsequent autoradiography showed radioactivity to be present primarily in amacrine cells. Under these conditions, most of the radioactivity contained in the retinas remained in the chemical form of GABA. Autoradiography and immunohistochemistry of alternate sections showed the amacrine cells that accumulate 3H-GABA to be the same cells that contain endogenous GABA immunoreactivity. These include the starburst cells, the indoleamine-accumulating cells, and other, as yet unidentified amacrine cells. The localization confirms previous immunohistochemical findings. When retinas containing 3H-GABA were expressed to elevated concentrations of K+, their content of 3H-GABA decreased. Autoradiography showed a reduced 3H-GABA content in all of the cells that contained 3H-GABA. Since those include the starburst cells, previously shown to be cholinergic, the finding demonstrates that the starburst cells release both ACh and GABA. Retinas simultaneously labeled with 14C-GABA and 3H-ACh were superfused, and the release of radioactive compounds from the retina was studied. Depolarization by elevated K+ caused an increased recovery of both ACh and GABA in the superfusate, but the predominant mechanisms of their release appeared to be different. The stimulated release of ACh was entirely Ca2+ dependent, while the release of radioactivity originating from GABA was much less so. A concentration-dependent counterflux (homoexchange) of intracellular GABA was demonstrated by raising the extracellular concentration of GABA (or nipecotic acid). These results suggest that a large outward flux of GABA occurs via the GABA transporter, probably by the potential-sensitive mechanism studied by Schwartz (1982, 1987). Stimulation of double-labeled retinas by flashing light or moving bars always increased the release of ACh, and the release was entirely dependent on the presence of extracellular Ca2+. Stimulation with light never caused a detectable release of GABA. This was unexpected, since the two neurotransmitters are present in the same amacrine cells: stimulation adequate to release one neurotransmitter should release both.(ABSTRACT TRUNCATED AT 400 WORDS)


Work, Care and Gender during the COVID-19 Crisis.
We explore the effects of the COVID-19 crisis and the associated restrictions to economic activity on paid and unpaid work for men and women in the United Kingdom. Using data from the COVID-19 supplement of Understanding Society, we find evidence that labour market outcomes of men and women were roughly equally affected at the extensive margin, as measured by the incidence of job loss or furloughing. But, if anything, women suffered smaller losses at the intensive margin, experiencing slightly smaller changes in hours and earnings. Within the household, women provided on average a larger share of increased childcare needs, but in an important share of households fathers became the primary childcare providers. These distributional consequences of the pandemic may be important to understand its inequality legacy over the longer term.


Macrophages: The Good, the Bad, and the Gluttony.
Macrophages are dynamic cells that play critical roles in the induction and resolution of sterile inflammation. In this review, we will compile and interpret recent findings on the plasticity of macrophages and how these cells contribute to the development of non-infectious inflammatory diseases, with a particular focus on allergic and autoimmune disorders. The critical roles of macrophages in the resolution of inflammation will then be examined, emphasizing the ability of macrophages to clear apoptotic immune cells. Rheumatoid arthritis (RA) is a chronic autoimmune-driven spectrum of diseases where persistent inflammation results in synovial hyperplasia and excessive immune cell accumulation, leading to remodeling and reduced function in affected joints. Macrophages are central to the pathophysiology of RA, driving episodic cycles of chronic inflammation and tissue destruction. RA patients have increased numbers of active M1 polarized pro-inflammatory macrophages and few or inactive M2 type cells. This imbalance in macrophage homeostasis is a main contributor to pro-inflammatory mediators in RA, resulting in continual activation of immune and stromal populations and accelerated tissue remodeling. Modulation of macrophage phenotype and function remains a key therapeutic goal for the treatment of this disease. Intriguingly, therapeutic intervention with glucocorticoids or other DMARDs promotes the re-polarization of M1 macrophages to an anti-inflammatory M2 phenotype; this reprogramming is dependent on metabolic changes to promote phenotypic switching. Allergic asthma is associated with Th2-polarised airway inflammation, structural remodeling of the large airways, and airway hyperresponsiveness. Macrophage polarization has a profound impact on asthma pathogenesis, as the response to allergen exposure is regulated by an intricate interplay between local immune factors including cytokines, chemokines and danger signals from neighboring cells. In the Th2-polarized environment characteristic of allergic asthma, high levels of IL-4 produced by locally infiltrating innate lymphoid cells and helper T cells promote the acquisition of an alternatively activated M2a phenotype in macrophages, with myriad effects on the local immune response and airway structure. Targeting regulators of macrophage plasticity is currently being pursued in the treatment of allergic asthma and other allergic diseases. Macrophages promote the re-balancing of pro-inflammatory responses towards pro-resolution responses and are thus central to the success of an inflammatory response. It has long been established that apoptosis supports monocyte and macrophage recruitment to sites of inflammation, facilitating subsequent corpse clearance. This drives resolution responses and mediates a phenotypic switch in the polarity of macrophages. However, the role of apoptotic cell-derived extracellular vesicles (ACdEV) in the recruitment and control of macrophage phenotype has received remarkably little attention. ACdEV are powerful mediators of intercellular communication, carrying a wealth of lipid and protein mediators that may modulate macrophage phenotype, including a cargo of active immune-modulating enzymes. The impact of such interactions may result in repair or disease in different contexts. In this review, we will discuss the origin, characterization, and activity of macrophages in sterile inflammatory diseases and the underlying mechanisms of macrophage polarization via ACdEV and apoptotic cell clearance, in order to provide new insights into therapeutic strategies that could exploit the capabilities of these agile and responsive cells.


Mutation history of the roma/gypsies.
The 8-10 million European Roma/Gypsies are a founder population of common origins that has subsequently split into multiple socially divergent and geographically dispersed Gypsy groups. Unlike other founder populations, whose genealogy has been extensively documented, the demographic history of the Gypsies is not fully understood and, given the lack of written records, has to be inferred from current genetic data. In this study, we have used five disease loci harboring private Gypsy mutations to examine some missing historical parameters and current structure. We analyzed the frequency distribution of the five mutations in 832-1,363 unrelated controls, representing 14 Gypsy populations, and the diversification of chromosomal haplotypes in 501 members of affected families. Sharing of mutations and high carrier rates supported a strong founder effect, and the identity of the congenital myasthenia 1267delG mutation in Gypsy and Indian/Pakistani chromosomes provided the best evidence yet of the Indian origins of the Gypsies. However, dramatic differences in mutation frequencies and haplotype divergence and very limited haplotype sharing pointed to strong internal differentiation and characterized the Gypsies as a founder population comprising multiple subisolates. Using disease haplotype coalescence times at the different loci, we estimated that the entire Gypsy population was founded approximately 32-40 generations ago, with secondary and tertiary founder events occurring approximately 16-25 generations ago. The existence of multiple subisolates, with endogamy maintained to the present day, suggests a general approach to complex disorders in which initial gene mapping could be performed in large families from a single Gypsy group, whereas fine mapping would rely on the informed sampling of the divergent subisolates and searching for the shared genomic region that displays the strongest linkage disequilibrium with the disease.


Study protocol: differential effects of diet and physical activity based interventions in pregnancy on maternal and fetal outcomes--individual patient data (IPD) meta-analysis and health economic evaluation.
Pregnant women who gain excess weight are at risk of complications during pregnancy and in the long term. Interventions based on diet and physical activity minimise gestational weight gain with varied effect on clinical outcomes. The effect of interventions on varied groups of women based on body mass index, age, ethnicity, socioeconomic status, parity, and underlying medical conditions is not clear. Our individual patient data (IPD) meta-analysis of randomised trials will assess the differential effect of diet- and physical activity-based interventions on maternal weight gain and pregnancy outcomes in clinically relevant subgroups of women. Randomised trials on diet and physical activity in pregnancy will be identified by searching the following databases: MEDLINE, EMBASE, BIOSIS, LILACS, Pascal, Science Citation Index, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effects, and Health Technology Assessment Database. Primary researchers of the identified trials are invited to join the International Weight Management in Pregnancy Collaborative Network and share their individual patient data. We will reanalyse each study separately and confirm the findings with the original authors. Then, for each intervention type and outcome, we will perform as appropriate either a one-step or a two-step IPD meta-analysis to obtain summary estimates of effects and 95% confidence intervals, for all women combined and for each subgroup of interest. The primary outcomes are gestational weight gain and composite adverse maternal and fetal outcomes. The difference in effects between subgroups will be estimated and between-study heterogeneity suitably quantified and explored. The potential for publication bias and availability bias in the IPD obtained will be investigated. We will conduct a model-based economic evaluation to assess the cost effectiveness of the interventions to manage weight gain in pregnancy and undertake a value of information analysis to inform future research. PROSPERO 2013: CRD42013003804.


Low-vision reading speed: influences of linguistic inference and aging.
Reading is a dynamic task involving both linguistic and visual analysis. In this study, we asked how two types of linguistic information--characters used in segmenting words from one another, and sentence context--differ in their usefulness for people with normal and low vision. Given evidence for age-related differences in some forms of cognitive processing, we also investigated the effect of age. There were four groups of 10 participants: vision status (normal, low) crossed with age (young, <35 years; old, >65 years). Reading speeds were compared for regularly spaced text and text in which the spaces were removed, a manipulation intended to eliminate local cues for text segmentation and force attention to clusters of letters or whole words. We also evaluated the effect of sentence context by comparing reading speeds for regular sentences and sentences in which word order was scrambled. Removal of spaces had a greater impact on low vision than normal vision, reducing average speeds to 45% and 66% of speeds for regularly spaced text, respectively. We interpret this to mean that people with low vision have less access to spatially distributed linguistic regularities of text such as prefixes, suffixes, or word length. Removal of sentence context through scrambling had a greater impact on normal vision than low vision, reducing mean reading speed to 53% and 66%, respectively. Finally, comparison of our young and old readers showed no major differences in the use of sentence context or in the impact of removing spaces between words. People with low vision appear to rely more on spacing information in sentences, whereas people with normal vision appear to make better use of sentence context, irrespective of age.


Antitumoral activity of interferon-gamma involved in impaired immune function in cancer patients.
Insufficient immunosurveillance is an important aspect in early tumorigenesis and in the pathogenesis of malignant disease. In the later course of cancer, the development of immunodeficiency is considered the major reason for disease progression and death. Within the anti-tumoral host defense reaction, Th1-type cytokine interferon-gamma (IFN-gamma) is of particular relevance. IFN-gamma stimulates several anti-proliferative and thus tumoricidal biochemical pathways in macrophages and other cells and also in tumor cell lines. These include inducible nitric oxide synthase, indoleamine (2, 3)-dioxygenase, an enzyme degrading the essential amino acid tryptophan, and the production of reactive oxygen species and neopterin in human macrophages and dendritic cells. Although the anti-proliferative strategy of the immune system aims to inhibit the growth of malignant cells, it can also affect T-cell response and thus contribute to the development of immunodeficiency. Accelerated degradation of tryptophan and increased production of neopterin were found to parallel the course of malignant diseases. Moreover, a higher degree of these metabolic changes characterizes poor prognosis and is associated with the development of anemia, weight loss and depressive mood in patients. Available data suggest that immunodeficiency in cancer patients may develop as a long-term side-effect of the antiproliferative and pro-apoptotic mechanisms elicited within Th1-type immune response, and enhanced production of pro-inflammatory cytokine IFN-gamma seems to be critically involved.


Fenfluramine, a serotonin-releasing drug, prevents seizure-induced respiratory arrest and is anticonvulsant in the DBA/1 mouse model of SUDEP.
Prevention of sudden unexpected death in epilepsy (SUDEP) is a critical goal for epilepsy therapy. The DBA/1 mouse model of SUDEP exhibits an elevated susceptibility to seizure-induced death in response to electroconvulsive shock, hyperthermia, convulsant drug, and acoustic stimulation. The serotonin hypothesis of SUDEP is based on findings that treatments which modify serotonergic function significantly alter susceptibility to seizure-induced sudden death in several epilepsy models, including DBA/1 mice. Serotonergic abnormalities have also recently been observed in human SUDEP. Fenfluramine is a drug that enhances serotonin release in the brain. Recent studies have found that the addition of fenfluramine improved seizure control in patients with Dravet syndrome, which has a high incidence of SUDEP. Therefore, we investigated the effects of fenfluramine on seizures and seizure-induced respiratory arrest (S-IRA) in DBA/1 mice. The dose and time course of the effects of fenfluramine (i.p.) on audiogenic seizures (Sz) induced by an electric bell in DBA/1 mice were determined. Videos of Sz-induced behaviors were recorded for analysis. Statistical significance (P < 0.05) was evaluated using the chi-square test. Sixteen hours after administration of 15 mg/kg of fenfluramine, a high incidence of selective block of S-IRA susceptibility (P < 0.001) occurred in DBA/1 mice without blocking any convulsive behavior. Thirty minutes after 20-40 mg/kg of fenfluramine, significant reductions of seizure incidence and severity, as well as S-IRA susceptibility occurred, which were long-lasting (≥48 hours). The median effective dose (ED50 ) of fenfluramine for significantly reducing Sz at 30 minutes was 21 mg/kg. This study presents the first evidence for the effectiveness of fenfluramine in reducing seizure incidence, severity, and S-IRA susceptibility in a mammalian SUDEP model. The ability of fenfluramine to block S-IRA selectively suggests the potential usefulness of fenfluramine in prophylaxis of SUDEP. These results further confirm and extend the serotonin hypothesis of SUDEP.


Antihyperalgesia induced by Heliopsis longipes extract.
Heliopsis longipes is an herbaceous plant found in Mexico. Heliopsis longipes is traditionally used for its analgesic and anesthetic properties. Plant extracts may represent a therapeutic advantage for the clinical treatment of pain. Therefore, the main objective of this study was to determine the possible antihyperalgesic effect produced by the Heliopsis longipes ethanolic extract (HLEE) in the Hargreaves model of thermal hyperalgesia in the mouse. HLEE was administrated systemically to mice and the antihyperalgesic effect was evaluated using the thermal hyperalgesia test. Oral Administration of HLEE produced a dose-dependent antihyperalgesic effect. Previously, it was reported that Heliopsis longipes extract was able to release GABA in mice temporal cortex slices. Therefore, it is likely that the antihyperalgesic effect observed in our study could result from GABA liberation and its inhibition of excessive excitation of nociceptive circuits in the thalamus and cortex evoked by tissue injury. Our results suggest that HLEE may represent a therapeutic advantage for the clinical treatment of inflammatory pain.


Frequent genes in rare diseases: panel-based next generation sequencing to disclose causal mutations in hereditary neuropathies.
Hereditary neuropathies comprise a wide variety of chronic diseases associated to more than 80 genes identified to date. We herein examined 612 index patients with either a Charcot-Marie-Tooth phenotype, hereditary sensory neuropathy, familial amyloid neuropathy, or small fiber neuropathy using a customized multigene panel based on the next generation sequencing technique. In 121 cases (19.8%), we identified at least one putative pathogenic mutation. Of these, 54.4% showed an autosomal dominant, 33.9% an autosomal recessive, and 11.6% an X-linked inheritance. The most frequently affected genes were PMP22 (16.4%), GJB1 (10.7%), MPZ, and SH3TC2 (both 9.9%), and MFN2 (8.3%). We further detected likely or known pathogenic variants in HINT1, HSPB1, NEFL, PRX, IGHMBP2, NDRG1, TTR, EGR2, FIG4, GDAP1, LMNA, LRSAM1, POLG, TRPV4, AARS, BIC2, DHTKD1, FGD4, HK1, INF2, KIF5A, PDK3, REEP1, SBF1, SBF2, SCN9A, and SPTLC2 with a declining frequency. Thirty-four novel variants were considered likely pathogenic not having previously been described in association with any disorder in the literature. In one patient, two homozygous mutations in HK1 were detected in the multigene panel, but not by whole exome sequencing. A novel missense mutation in KIF5A was considered pathogenic because of the highly compatible phenotype. In one patient, the plasma sphingolipid profile could functionally prove the pathogenicity of a mutation in SPTLC2. One pathogenic mutation in MPZ was identified after being previously missed by Sanger sequencing. We conclude that panel based next generation sequencing is a useful, time- and cost-effective approach to assist clinicians in identifying the correct diagnosis and enable causative treatment considerations.


Tyrosinase overexpression promotes ATM-dependent p53 phosphorylation by quercetin and sensitizes melanoma cells to dacarbazine.
Dacarbazine (DTIC) has been used for the treatment of melanoma for decades. However, monotherapy with this chemotherapeutic agent results only in moderate response rates. To improve tumor response to DTIC current clinical trials in melanoma focus on combining a novel targeted agent with chemotherapy. Here, we demonstrate that tyrosinase which is commonly overexpressed in melanoma activates the bioflavonoid quercetin (Qct) and promotes an ataxia telangiectasia mutated (ATM)-dependent DNA damage response. This response sensitizes melanoma cells that overexpress tyrosinase to DTIC. In DB-1 melanoma cells that overexpress tyrosinase (Tyr(+) cells), the threshold for phosphorylation of ATM and p53 at serine 15 was observed at a low dose of Qct (25 microM) when compared to the mock transfected pcDNA3 cells, which required a higher dose (75 microM). Both pcDNA3 and Tyr(+) DB-1 cells demonstrated similar increases in phosphorylation of p53 at other serine sites, but in the Tyr(+) cells, DNApk expression was found to be reduced compared to control cells, indicating a shift towards an ATM-mediated response. The DB-1 control cells were resistant to DTIC, but were sensitized to apoptosis with high dose Qct, while Tyr(+) cells were sensitized to DTIC with low or high dose Qct. Qct also sensitized SK Mel 5 (p53 wildtype) and 28 (p53 mutant) cells to DTIC. However, when SK Mel 5 cells were transiently transfected with tyrosinase and treated with Qct plus DTIC, SK Mel 5 cells demonstrated a more than additive induction of apoptosis. Therefore, this study demonstrates that tyrosinase overexpression promotes an ATM-dependent p53 phosphorylation by Qct treatment and sensitizes melanoma cells to dacarbazine. In conclusion, these results suggest that Qct or Qct analogues may significantly improve DTIC response rates in tumors that express tyrosinase.


Mechanisms of action of mesenchymal stem cells in cardiac repair: potential influences on the cardiac stem cell niche.
Clinical and basic studies of cell-based myocardial therapy have proceeded at a rapid pace. Cell therapy could lead to successful cardiac regeneration or repair by any of three general mechanisms: differentiation of the administered cells into all of the cellular constituents of the heart; release of factors capable of paracrine signaling from the administered cells; and fusion of the administered cells with the existing constituents of the heart. Here, we argue that a fourth general mechanism could be operative: stimulation of endogenous repair by injected cells, which and might cause the regeneration of stem cell niches. In a porcine model of myocardial infarction, allogeneic mesenchymal stem cells stimulated substantial improvement in the ejection fraction, reduction of infarct size, and the growth of a rim of new cardiac tissue in the region in which the mesenchymal stem cells were injected. These effects occurred in the absence of definitive cardiac myocyte differentiation. After myocardial infarction, porcine hearts exhibit evidence of cardiac myocytes that have entered the cell cycle, neovascularization, and reduced levels of apoptosis. These data, in addition to new insights regarding the presence of endogenous cardiac stem cells, strongly support the concept that the heart could contain stem cell niches. Effective cell therapy could lead to restoration of these niches through multifaceted cell-cell interactions.


Supporting prison nurses: an action research approach to education.
Since April 2006, commissioning responsibility for healthcare services in public prisons has been fully devolved to NHS primary care trusts (PCTs), with the expectation that offenders will have access to the same range and quality of health services available to the wider population. In order to support prison nurses in meeting this goal, a PCT and university established a partnership, which used an action research approach to develop, instigate and evaluate a bespoke educational programme for nurses working in two local prisons. This article outlines the processes involved in the design and implementation of the programme. It also reports on findings from pre- and post-intervention questionnaires and focus groups with course participants, and semi-structured interviews with key stakeholders, which suggest that the innovation had a positive impact on the nurses' confidence, assertiveness, clinical expertise and approach to change. The article concludes that the action research project should continue, but its scope should now broaden to address educational support for healthcare assistants, collaborative learning between prison officers and prison nurses, and the implementation of clinical supervision and action learning sets.


Etching and printing of diffractive optical microstructures by a femtosecond excimer laser.
Diffractive optics fabrication is performed by two complementary processing methods that rely on the photoablation of materials by ultrashort UV laser pulses. The spatially selective ablation of materials permits the direct microetching of high-quality surface-relief patterns. In addition, the direct, spatially selective transfer of the ablated material onto planar and nonplanar receiving substrates provides a complementary microprinting operation. The radiation from the ultrashort pulsed excimer laser results in superior quality at relatively low-energy density levels, owing to the short absorption length and minimal thermal-diffusion effects. Computer-generated holographic structures are produced by both modes of operation. Submicrometer features, including Bragg-type structures, are microprinted onto planar and high-curvature optical-fiber surfaces, demonstrating the unique ability of the schemes for complex microstructure and potentially nanostructure development.


Time-varying social support and time to death in the cardiovascular health study.
There is a consensus that social connectedness is integral for a long, healthy life. However, studies of social support and survival have primarily relied on baseline social support measures, potentially missing the effects of fluctuations of perceived support over time. This is especially important for older adults who experience increased changes in disability. This study examined whether among older adults time-varying perceived social support was associated with time to death (main effect model of support) and whether time-varying disability was a modifier (stress-buffering model of support). Gender and marital status were also examined as modifiers. Older adults in the Cardiovascular Health Study (N = 5,201) completed self- report measures of demographics and psychological health and clinical risk factors for mortality at baseline (1989-1990). Perceived social support and disability were measured from baseline through Wave 11 (1998-1999). Cox regression of time to death with time-varying covariates was performed. Time-varying as well as baseline-only perceived social support was associated with greater survival in the unadjusted models but not after adjustment. Gender, marital status, and time-varying disability were not significant modifiers. In contrast with the previously reported association between baseline individual differences in perceived social support and time to death, older adults' baseline-only and fluctuating perceptions of perceived support over time were not associated with time to death after adjustment for other clinical physical and psychological risk factors. Research is needed to identify other relationship factors that may be more informative as time-varying predictors of health and longevity in large longitudinal data sets. (PsycINFO Database Record (c) 2018 APA, all rights reserved).