Weak and uneven associations of home, neighborhood, and school environments with stress hormone output across multiple timescales.
The progression of lifelong trajectories of socioeconomic inequalities in health and mortality begins in childhood. Dysregulation in cortisol, a stress hormone that is the primary output of the hypothalamus-pituitary-adrenal (HPA) axis, has been hypothesized to be a mechanism for how early environmental adversity compromises health. However, despite the popularity of cortisol as a biomarker for stress and adversity, little is known about whether cortisol output differs in children being raised in socioeconomically disadvantaged environments. Here, we show that there are few differences between advantaged and disadvantaged children in their cortisol output. In 8-14-year-old children from the population-based Texas Twin Project, we measured cortisol output at three different timescales: (a) diurnal fluctuation in salivary cortisol (n = 400), (b) salivary cortisol reactivity and recovery after exposure to the Trier Social Stress Test (n = 444), and (c) cortisol concentration in hair (n = 1210). These measures converged on two moderately correlated, yet distinguishable, dimensions of HPA function. We tested differences in cortisol output across nine aspects of social disadvantage at the home (e.g., family socioeconomic status), school (e.g., average levels of academic achievement), and neighborhood (e.g., concentrated poverty). Children living in neighborhoods with higher concentrated poverty had higher diurnal cortisol output, as measured in saliva; otherwise, child cortisol output was unrelated to any other aspect of social disadvantage. Overall, we find limited support for alteration in HPA axis functioning as a general mechanism for the health consequences of socioeconomic inequality in childhood.


An ecosystem modelling approach for deriving water quality criteria.
Ecological effects of chemicals on ecosystems are the result of direct effects of the chemical, determined in single-species toxicity testing, and indirect effects due to ecological interactions between species. Current experimental methods to account for such interactions are expensive. Hence, mathematical models of ecosystems have been proposed as an alternative. The use of these models often requires extensive calibration, which hampers their use as a general tool in ecological effect assessments. Here we present a novel ecosystem modelling approach which assesses effects of chemicals on ecosystems by integrating single-species toxicity test results and ecological interactions, without the need for calibration on case-specific data. The methodology is validated by comparing predicted ecological effects of copper in a freshwater planktonic ecosystem with an experimental ecosystem data set. Two main effects reflected by this data set (a decrease of cladocerans and an increase of small phytoplankton) which were unpredictable from single-species toxicity test results alone, were predicted accurately by the developed model. Effects on populations which don't interact directly with other populations, were predicted equally well by single-species toxicity test results as by the ecosystem model. The small amount of required data and the high predictive capacity can make this ecosystem modelling approach an efficient tool in water quality criteria derivation for chemicals.


Fluoromodule-based reporter/probes designed for in vivo fluorescence imaging.
Optical imaging of whole, living animals has proven to be a powerful tool in multiple areas of preclinical research and has allowed noninvasive monitoring of immune responses, tumor and pathogen growth, and treatment responses in longitudinal studies. However, fluorescence-based studies in animals are challenging because tissue absorbs and autofluoresces strongly in the visible light spectrum. These optical properties drive development and use of fluorescent labels that absorb and emit at longer wavelengths. Here, we present a far-red absorbing fluoromodule-based reporter/probe system and show that this system can be used for imaging in living mice. The probe we developed is a fluorogenic dye called SC1 that is dark in solution but highly fluorescent when bound to its cognate reporter, Mars1. The reporter/probe complex, or fluoromodule, produced peak emission near 730 nm. Mars1 was able to bind a variety of structurally similar probes that differ in color and membrane permeability. We demonstrated that a tool kit of multiple probes can be used to label extracellular and intracellular reporter-tagged receptor pools with 2 colors. Imaging studies may benefit from this far-red excited reporter/probe system, which features tight coupling between probe fluorescence and reporter binding and offers the option of using an expandable family of fluorogenic probes with a single reporter gene.


Prediction of adult skinfolds and body mass from infancy through adolescence.
In this paper age-to-age correlations for the body mass index and for skinfolds are evaluated for a sample of normal children studied from birth to adulthood. While correlations over larger age spans are modest, they become appreciable from childhood to adolescence and from adolescence to adulthood. Correlations are consistently higher for boys compared to girls, and only for the former does the body mass index correlate better than skinfolds. Significant correlations between weight increase in the first year and the adult body mass index were found, as well as between the age of 'adiposity rebound' and the adult body mass index. However, the small size of the correlations forbids any predictive applications. As it turns out, the individual prediction of the adult size of the body mass index or of skinfolds is a thorny problem, whatever variables and methods are chosen. The precision of such a prediction is very low up to late childhood and becomes somewhat better in adolescence. From a positive side, this leaves much room for overweight children to improve their state. On the other hand, the relative risk for becoming a heavy adult is much increased for those who are already heavy as children and adolescents. This underlines the dangers of early overweight from an epidemiological viewpoint.


Cysteine 98 in CYP3A4 contributes to conformational integrity required for P450 interaction with CYP reductase.
Previously human cytochrome P450 3A4 was efficiently and specifically photolabeled by the photoaffinity ligand lapachenole. One of the modification sites was identified as cysteine 98 in the B-C loop region of the protein [B. Wen, C.E. Doneanu, C.A. Gartner, A.G. Roberts, W.M. Atkins, S.D. Nelson, Biochemistry 44 (2005) 1833-1845]. Loss of CO binding capacity and subsequent decrease of catalytic activity were observed in the labeled CYP3A4, which suggested that aromatic substitution on residue 98 triggered a critical conformational change and subsequent loss of enzyme activity. To test this hypothesis, C98A, C98S, C98F, and C98W mutants were generated by site-directed mutagenesis and expressed functionally as oligohistidine-tagged proteins. Unlike the mono-adduction observed in the wild-type protein, simultaneous multiple adductions occurred when C98F and C98W were photolabeled under the same conditions as the wild-type enzyme, indicating a substantial conformational change in these two mutants compared with the wild-type protein. Kinetic analysis revealed that the C98W mutant had a drastic 16-fold decrease in catalytic efficiency (V(max)/K(m)) for 1'-OH midazolam formation, and about an 8-fold decrease in catalytic efficiency (V(max)/K(m)) for 4-OH midazolam formation, while the C98A and C98S mutants retained the same enzyme activity as the wild-type enzyme. Photolabeling of C98A and C98S with lapachenole resulted in monoadduction of only Cys-468, in contrast to the labeling of Cys-98 in wild-type CYP3A4, demonstrating the marked selectivity of this photoaffinity ligand for cysteine residues. The slight increases in the midazolam binding constants (K(s)) in these mutants suggested negligible perturbation of the heme environment. Further activity studies using different P450:reductase ratios suggested that the affinity of P450 to reductase was significantly decreased in the C98W mutant, but not in the C98A and C98S mutants. In addition, the C98W mutant exhibited a 41% decrease in the maximum electron flow rate between P450 and reductase as measured by reduced nicotinamide adenine dinucleotide phosphate consumption at a saturating reductase concentration. In conclusion, our data strongly suggest that cysteine 98 in the B-C loop region significantly contributes to conformational integrity and catalytic activity of CYP3A4, and that this residue or residues nearby might be involved in an interaction with P450 reductase.


Living inside the box: environmental effects on mouse models of human disease.
The impact of the laboratory environment on animal models of human disease, particularly the mouse, has recently come under intense scrutiny regarding both the reproducibility of such environments and their ability to accurately recapitulate elements of human environmental conditions. One common objection to the use of mice in highly controlled facilities is that humans live in much more diverse and stressful environments, which affects the expression and characteristics of disease phenotypes. In this Special Article, we review some of the known effects of the laboratory environment on mouse phenotypes and compare them with environmental effects on humans that modify phenotypes or, in some cases, have driven genetic adaptation. We conclude that the 'boxes' inhabited by mice and humans have much in common, but that, when attempting to tease out the effects of environment on phenotype, a controlled and, importantly, well-characterized environment is essential.


'You see the empty bed which means it's either a transplant or a death': a qualitative study exploring the impact of death in the haemodialysis community.
To explore the impact of the death of a patient in the haemodialysis unit on fellow patients. We interviewed patients on dialysis in a tertiary dialysis centre using semistructured interviews. We purposively sampled patients who had experienced the death of a fellow patient. After interviews were transcribed, they were thematically analysed by independent members of the research team using inductive analysis. Input from the team during analysis ensured the rigour and quality of the findings. 10 participants completed the interviews (6 females and 4 males with an age range of 42-88 years). The four core themes that emerged from the interviews included: (1) patients' relationship to haemodialysis, (2) how patients define the haemodialysis community, (3) patients' views on death and bereavement and (4) patients' expectations around death in the dialysis community. Patients noticed avoidance behaviour by staff in relation to discussing death in the unit and would prefer a culture of open acknowledgement. Staff acknowledgement of death is of central importance to patients on haemodialysis who feel that the staff are part of their community. This should guide the development of appropriate bereavement support services and a framework that promotes the provision of guidance for staff and patients in this unique clinical setting. However, the authors acknowledge the homogenous sample recruited in a single setting may limit the transferability of the study. Further work is needed to understand diverse patient and nurse experiences and perceptions when sharing the knowledge of a patient's death and how they react to loss.


Bioaccumulation and toxicokinetics of 42 polychlorinated biphenyl congeners in American kestrels (Falco sparverius).
The bioaccumulation and toxicokinetics of 42 polychlorinated biphenyls (PCBs) was determined in male American kestrels exposed to an Aroclor-contaminated diet for 120 d followed by a 348-d depuration period. The birds were housed under ambient outdoor temperatures to permit normal fluctuations in body weight during the study. Whole body PCB clearance, plasma/fat distribution coefficients, and plasma PCB clearance constants were determined for individual PCBs to calibrate a two-compartment rate constant model in order to describe PCB elimination in the birds. Plasma/fat partition coefficients (K(PF)) averaged 0.0060 +/- 0.0001 for all congeners of study, were not dependent on chemical hydrophobicity, and did not change in summer versus winter sacrificed animals. Plasma clearance constants (k'pc) for PCBs were observed to be dependent on both chlorine substitution patterns and congener hydrophobicity. Polychlorinated biphenyl congeners categorized as readily cleared congeners contained vicinal meta-para hydrogen substituents on at least one phenyl ring, while slowly cleared congeners were chlorine hindered at these positions. A general equation was derived to predict plasma clearance constants for all tri- to octachlorobiphenyls based on the presence of an open meta-para site on one of the phenyl rings and from the n-octanol-water partition coefficient of the chemical. The equation was validated by comparing predicted versus measured relative biomagnification factors of PCBs determined in birds at the end of the dosing period. The two-compartment model calibrated for PCB elimination in American kestrels may be used to describe PCB toxicokinetics in wild birds provided that seasonal fluctuations in the fat content of the modeled population is known.


Mediators between bereavement and somatic symptoms.
In our research we examined the frequency of somatic symptoms among bereaved (N = 185) and non-bereaved men and women in a national representative sample (N = 4041) and investigated the possible mediating factors between bereavement status and somatic symptoms. Somatic symptoms were measured by the Patient Health Questionnaire (PHQ-15), anxiety with a four-point anxiety rating scale, and depression with a nine-item shortened version of the Beck Depression Inventory. Among the bereaved, somatic symptoms proved to be significantly more frequent in both genders when compared to the non-bereaved, as did anxiety and depression. On the multivariate level, the results show that both anxiety and depression proved to be a mediator between somatic symptoms and bereavement. The effect sizes indicated that for both genders, anxiety was a stronger predictor of somatic symptoms than depression. The results of our research indicate that somatic symptoms accompanying bereavement are not direct consequences of this state but they can be traced back to the associated anxiety and depression. These results draw attention to the need to recognize anxiety and depression looming in the background of somatic complaints in bereavement and to the importance of the dissemination of related information.


Adoption and impact of improved cookstoves in Lambayeque, Peru, 2017.
Approximately three billion people in Asia, Africa, and the Americas cook with biomass, cleaner cooking technologies with the potential to reduce household air pollution exposure. It is necessary to assess the adoption and long-term use of these stoves, measure perceived benefits among users, and use this information to provide feedback to programs that are implementing new cooking technologies. The aim of this study is to determine the level of adoption and impact of improved biomass cookstoves in the rural area of Lambayeque, Peru, in 2017. A descriptive cross-sectional study was conducted. A total of 52 homes were surveyed in the districts of Pacora and Íllimo where improved biomass stoves were introduced between 2005 and 2013. A questionnaire for the assessment of adoption and impact indexes proposed by the Global Alliance for Clean Cookstoves was applied. The STROBE checklist was used. The mean number of years with the improved biomass cookstove was 9.1 (standard deviation: 2.9); 51.9% always used the improved stove, and 34.6% never used it due to destruction during the El Niño phenomenon in 2017. The median impact index was 5.62; 19.2% had a very good/good adoption. The median adoption index was 6.5; 25% had a very high/high impact. The use of the traditional or open fire biomass stove persisted in 61.5% of the houses. The adoption and impact of improved biomass cookstoves were acceptable, but traditional stove use persisted in more than half of the houses. Households used a mix of different stove technologies. Gas stoves were used more frequently for breakfast or dinner, while the traditional biomass stoves were used for larger lunchtime meals.


MetaPrism: A versatile toolkit for joint taxa/gene analysis of metagenomic sequencing data.
In microbiome research, metagenomic sequencing generates enormous amounts of data. These data are typically classified into taxa for taxonomy analysis, or into genes for functional analysis. However, a joint analysis where the reads are classified into taxa-specific genes is often overlooked. To enable the analysis of this biologically meaningful feature, we developed a novel bioinformatic toolkit, MetaPrism, which can analyze sequence reads for a set of joint taxa/gene analyses to: 1) classify sequence reads and estimate the abundances for taxa-specific genes; 2) tabularize and visualize taxa-specific gene abundances; 3) compare the abundances between groups; and 4) build prediction models for clinical outcome. We illustrated these functions using a published microbiome metagenomics dataset from patients treated with immune checkpoint inhibitor therapy and showed the joint features can serve as potential biomarkers to predict therapeutic responses. MetaPrism is a toolkit for joint taxa and gene analysis. It offers biological insights on the taxa-specific genes on top of the taxa-alone or gene-alone analysis. MetaPrism is open-source software and freely available at https://github.com/jiwoongbio/MetaPrism. The example script to reproduce the manuscript is also provided in the above code repository.


Mammalian Elongin A complex mediates DNA-damage-induced ubiquitylation and degradation of Rpb1.
The Elongin complex stimulates the rate of transcription elongation by RNA polymerase II (pol II) by suppressing transient pausing of the pol II at many sites along the DNA. Elongin is composed of a transcriptionally active A subunit and two small regulatory B and C subunits, which can form an isolable Elongin BC subcomplex. Here, we have shown that both the ubiquitylation and proteasomal degradation of the largest subunit of pol II (Rpb1) following UV-irradiation are significantly suppressed in Elongin A-deficient cells; however, in both cases suppression is rescued by transfection of wild-type Elongin A. Moreover, we have demonstrated that the Elongin A-Elongin BC complex is capable of assembling with the Cul5/Rbx2 module, and that this hetero-pentamer complex efficiently ubiquitylates Rpb1 in vitro. Mechanistic studies indicate that colocalization of Elongin A and Cul5 in cells and the interaction of Elongin A with the Ser5-phosphorylated form of Rpb1 are strongly enhanced following UV-irradiation. Taken together, our results suggest that mammalian Elongin A is directly involved in ubiquitylation and degradation of Rpb1 following DNA damage.


Insomnia and changes in alcohol consumption: Relation between possible complicated grief and depression among bereaved family caregivers.
Bereaved individuals are known to have greater health risks, such as insomnia, excessive alcohol intake, and depression. However, few studies have investigated the relation between these risks and bereavement outcomes, namely complicated grief (CG) and major depressive disorder (MDD). This study aimed to assess the relation between insomnia, changes in alcohol consumption, and CG or MDD. A cross-sectional, self-report questionnaire survey was conducted between May and July 2014 on 20 acute hospitals, 133 inpatient palliative care units (PCUs), and 22 home hospice services. Questionnaires were sent to the bereaved family members identified by each institution. Data were obtained from 814, 7,291, and 1,018 family members from acute hospitals, PCUs, and home hospice services, respectively. Significant associations were found between CG or MDD and all sleep condition symptoms (OR: 1.48-12.88; all p<0.0001) and between changes in alcohol intake (OR: 1.63-3.55; all p<0.0001). The majority were family members who had lost a loved one to cancer in a PCU, the psychological health of nonresponders was unavailable, the results were based on self-report data, and no clinical assessment interviews were done; this could limit the generalizability of the findings. Overall, 14% and 17% of the respondents reported increased and decreased alcohol consumption, respectively, and 46-61% reported experiencing insomnia. Interestingly, both increase and decrease in alcohol intake after bereavement were risk factors of possible CG or MDD. These results suggest that assessing sleep conditions and alcohol consumption might help prevent severe psychological impairments in bereaved individuals.


Past exposure to vaccines and subsequent risk of Alzheimer's disease.
It has been suggested that changes to the immune system could be a factor in age-related conditions such as Alzheimer's disease. Our objective was to examine the association between past exposure to conventional vaccines and risk of Alzheimer's disease. We analyzed data from a representative community sample of subjects 65 years of age or older participating in the Canadian Study of Health and Aging, a prospective cohort study of dementia. Screening and clinical evaluations were done at both baseline and follow-up. Past exposure to vaccines was assessed at baseline by means of a self-administered questionnaire. Of the 4392 eligible subjects who were cognitively unimpaired and for whom vaccine information was available at baseline (in 1991-1992) and who completed follow-up 5 years later (in 1996-1997), 527 were diagnosed as having cognitive impairment or dementia other than Alzheimer's disease and were excluded from these analyses. Of the remaining subjects, 3682 were cognitively unimpaired at follow-up and 183 were newly diagnosed as having Alzheimer's disease. After adjustment for age, sex and education, past exposure to vaccines against diphtheria or tetanus, poliomyelitis and influenza was associated with lower risk for Alzheimer's disease (odds ratio [OR] 0.41, 95% confidence interval [CI] 0.27-0.62; OR 0.60, 95% CI 0.37-0.99; and OR 0.75, 95% CI 0.54-1.04 respectively) than no exposure to these vaccines. Past exposure to vaccines against diphtheria or tetanus, poliomyelitis and influenza may protect against subsequent development of Alzheimer's disease.


Blood biomarker levels by total sleep duration: cross-sectional analyses in UK Biobank.
Short or long sleep duration has been associated with some major chronic diseases, but whether disease-related blood biomarkers vary according to habitual sleep duration is unclear. This cross-sectional study aimed to assess blood biomarker levels in relation to total sleep duration. The analysis includes 459,796 white British adults aged 40-69 during 2006-2010 in UK Biobank. At recruitment, blood samples and self-reported information on total sleep duration were collected from participants. A panel of blood biomarkers were measured. Using linear regression, we estimated geometric mean concentrations of blood biomarkers and mean ratio of ApoB/ApoA1 by sleep duration adjusted for sex, age at data collection, time of blood collection, and lifestyle covariates. Percentage differences in the concentrations of most biomarkers by sleep duration were modest. The largest differences were for C-reactive protein (CRP, an inflammatory biomarker) and gamma glutamyltransferase (GGT, a liver function biomarker), and the differences were markedly attenuated after multivariable-adjustment. The multivariable-adjusted geometric means of CRP and of GGT were 14% and 14% higher in <6 h vs 7-8 h of sleep; and 22% and 12% higher in >9 h vs 7-8 h of sleep, respectively. In white British adults, most blood biomarker levels varied only modestly with sleep duration and the remaining associations may be due to residual confounding.


Subpopulations of neurons in lOFC encode previous and current rewards at time of choice.
Studies of neural dynamics in lateral orbitofrontal cortex (lOFC) have shown that subsets of neurons that encode distinct aspects of behavior, such as value, may project to common downstream targets. However, it is unclear whether reward history, which may subserve lOFC's well-documented role in learning, is represented by functional subpopulations in lOFC. Previously, we analyzed neural recordings from rats performing a value-based decision-making task, and we documented trial-by-trial learning that required lOFC (Constantinople et al., 2019). Here, we characterize functional subpopulations of lOFC neurons during behavior, including their encoding of task variables. We found five distinct clusters of lOFC neurons, either based on clustering of their trial-averaged peristimulus time histograms (PSTHs), or a feature space defined by their average conditional firing rates aligned to different task variables. We observed weak encoding of reward attributes, but stronger encoding of reward history, the animal's left or right choice, and reward receipt across all clusters. Only one cluster, however, encoded the animal's reward history at the time shortly preceding the choice, suggesting a possible role in integrating previous and current trial outcomes at the time of choice. This cluster also exhibits qualitatively similar responses to identified corticostriatal projection neurons in a recent study (Hirokawa et al., 2019), and suggests a possible role for subpopulations of lOFC neurons in mediating trial-by-trial learning.


Chemical amplification accelerates reactive oxygen species triggered polymeric degradation.
Chemical amplification is a known strategy for improving the sensitivity of stimuli-responsive polymers. However, the chemical amplification effect has never been fully examined. Many questions remain about its mechanism and efficacy, obstructing its further implementation. Here, we design and demonstrate a reactive oxygen species (ROS) responsive polymer (ROS-ARP) with a chemical amplification strategy to dismiss these concerns. The ROS-ARP is designed to change the hydrophilicity by ROS, revealing a carboxylic acid, which also catalyzes ketal hydrolysis along the polymer backbone. The chemical amplification strategy of ROS-ARP accelerated the polymer degradation up to 17 fold compared to a previously reported ROS-responsive polymer. To investigate the mechanism behind this increased acceleration, we compared the degradation kinetics in various environments. Additionally, other effects such as hydrophilicity changes were excluded. The accelerated degradation of ROS-ARP is evaluated as a potential drug delivery system, demonstrating on-demand cargo release from the formulated polymeric particles.


Self-assembling, reactivity and molecular dynamics of fullerenol nanoparticles.
In this work structuring of water and insight into intermolecular interactions between water and fullerenol are studied throughout the process of forming nanoagglomerates at different temperatures applying both experimental and computational approaches. The obtained fullerenol nanoparticles (FNPs) are firstly characterized using dynamic light scattering, atomic force microscopy and transmission electron microscopy. The density, electrical conductivity and dynamic viscosity of aqueous fullerenol solutions are measured in the temperature range of 293.15 to 315.15 K. From the experimental density results other important thermodynamic values, such as apparent molar volumes and the partial molar volumes of water and fullerenol, are also calculated. To support the conclusion derived from the experimental density and calculated volumetric parameters, and to better understand the nature of the interactions with water, molecular dynamics simulations and radial distribution functions are also employed.


Next generation sequencing reveals genome downsizing in allotetraploid Nicotiana tabacum, predominantly through the elimination of paternally derived repetitive DNAs.
We used next generation sequencing to characterize and compare the genomes of the recently derived allotetraploid, Nicotiana tabacum (<200,000 years old), with its diploid progenitors, Nicotiana sylvestris (maternal, S-genome donor), and Nicotiana tomentosiformis (paternal, T-genome donor). Analysis of 14,634 repetitive DNA sequences in the genomes of the progenitor species and N. tabacum reveal all major types of retroelements found in angiosperms (genome proportions range between 17-22.5% and 2.3-3.5% for Ty3-gypsy elements and Ty1-copia elements, respectively). The diploid N. sylvestris genome exhibits evidence of recent bursts of sequence amplification and/or homogenization, whereas the genome of N. tomentosiformis lacks this signature and has considerably fewer homogenous repeats. In the derived allotetraploid N. tabacum, there is evidence of genome downsizing and sequences loss across most repeat types. This is particularly evident amongst the Ty3-gypsy retroelements in which all families identified are underrepresented in N. tabacum, as is 35S ribosomal DNA. Analysis of all repetitive DNA sequences indicates the T-genome of N. tabacum has experienced greater sequence loss than the S-genome, revealing preferential loss of paternally derived repetitive DNAs at a genome-wide level. Thus, the three genomes of N. sylvestris, N. tomentosiformis, and N. tabacum have experienced different evolutionary trajectories, with genomes that are dynamic, stable, and downsized, respectively.


Retroperitoneal unicentric Castleman's disease with multiple lymph node involvement.
A 78-year-old woman with B-symptoms was referred for a left adrenal incidentaloma of 5 cm. Imaging revealed features compatible with adrenal cancer. The authors excluded excess production of catecholamines or adrenal steroids. The tumour was removed by en bloc radical left retroperitonectomy with adrenalectomy, nephrectomy, interaortocaval lymphadenectomy and splenectomy. Histology demonstrated periadrenal hyaline vascular Castleman's disease with local infiltration and 14 positive lymph nodes. The lymphoid infiltrate spilled into the adjacent renal cortex. HHV8 was negative. The Ki67 proliferative index was 30-40% in germinal centres. There was no syn- or metachronous disease on extended imaging including fluorodeoxyglucose positron emission tomography-CT and narrow follow-up at 3 years. This is a rare case of unicentric hyaline vascular Castleman's disease with documented locoregional lymph node involvement. The case exemplifies the transition from unifocal unicentric disease into disseminated disease with involvement of multiple lymph node stations (multicentric disease). The authors demonstrate surgical cure by oncological resection.


Automatic identification and normalization of dosage forms in drug monographs.
Each day, millions of health consumers seek drug-related information on the Web. Despite some efforts in linking related resources, drug information is largely scattered in a wide variety of websites of different quality and credibility. As a step toward providing users with integrated access to multiple trustworthy drug resources, we aim to develop a method capable of identifying drug's dosage form information in addition to drug name recognition. We developed rules and patterns for identifying dosage forms from different sections of full-text drug monographs, and subsequently normalized them to standardized RxNorm dosage forms. Our method represents a significant improvement compared with a baseline lookup approach, achieving overall macro-averaged Precision of 80%, Recall of 98%, and F-Measure of 85%. We successfully developed an automatic approach for drug dosage form identification, which is critical for building links between different drug-related resources.


Mating behavior: facilitation in the female rat after cortical application of potassium chloride.
Mating behavior in ovariectomized female rats treated with estrogen can be greatly enhanced by subcutaneous injections of progesterone. Application of potassium chloride to the cortex of females previously treated with estrogen can also induce greatly increased sexual receptivity as indicated by the lordosis response. This facilitation of mating behavior by a treatment known to cause functional decortication suggests that mechanisms mediating female mating behavior are under tonic inhibition by an inhibitory system which involves the cerebral cortex.


Dominant component analysis of electrophysiological connectivity networks.
Connectivity matrices obtained from various modalities (DTI, MEG and fMRI) provide a unique insight into brain processes. Their high dimensionality necessitates the development of methods for population-based statistics, in the face of small sample sizes. In this paper, we present such a method applicable to functional connectivity networks, based on identifying the basis of dominant connectivity components that characterize the patterns of brain pathology and population variation. Projection of individual connectivity matrices into this basis allows for dimensionality reduction, facilitating subsequent statistical analysis. We find dominant components for a collection of connectivity matrices by using the projective non-negative component analysis technique which ensures that the components have non-negative elements and are non-negatively combined to obtain individual subject networks, facilitating interpretation. We demonstrate the feasibility of our novel framework by applying it to simulated connectivity matrices as well as to a clinical study using connectivity matrices derived from resting state magnetoencephalography (MEG) data in a population of subjects diagnosed with autism spectrum disorder (ASD).


Genomic variation in microbial populations inhabiting the marine subseafloor at deep-sea hydrothermal vents.
Little is known about evolutionary drivers of microbial populations in the warm subseafloor of deep-sea hydrothermal vents. Here we reconstruct 73 metagenome-assembled genomes (MAGs) from two geochemically distinct vent fields in the Mid-Cayman Rise to investigate patterns of genomic variation within subseafloor populations. Low-abundance populations with high intra-population diversity coexist alongside high-abundance populations with low genomic diversity, with taxonomic differences in patterns of genomic variation between the mafic Piccard and ultramafic Von Damm vent fields. Populations from Piccard are significantly enriched in nonsynonymous mutations, suggesting stronger purifying selection in Von Damm relative to Piccard. Comparison of nine Sulfurovum MAGs reveals two high-coverage, low-diversity MAGs from Piccard enriched in unique genes related to the cellular membrane, suggesting these populations were subject to distinct evolutionary pressures that may correlate with genes related to nutrient uptake, biofilm formation, or viral invasion. These results are consistent with distinct evolutionary histories between geochemically different vent fields, with implications for understanding evolutionary processes in subseafloor microbial populations.


A Simple Alternative Method for Preservation of 2-Methylisoborneol in Water Samples.
2-Methylisoborneol (2-MIB) is one of the most commonly observed taste and odor (T&O) compounds present in drinking water sources. As it is biodegradable, a preservation agent, typically mercury chloride, is needed if the water is not analyzed right after sampling. Since mercury is a toxic metal, an alternative chemical that is cheaper and less toxic is desirable. In this study, two chemicals commonly used in water treatment processes, chlorine (as sodium hypochlorite) and KMnO₄ (potassium permanganate), are studied to determine their feasibility as preservation agents for 2-MIB in water. Preservation experiments were first conducted in deionized water spiked with 2-MIB and with chlorine or permanganate at 4 and 25 °C. The results indicate that 2-MIB concentrations in the water samples spiked with both chemicals remained almost constant within 14 days for all the tested conditions, suggesting that oxidation and volatilization did not cause the loss of 2-MIB in the system. The experiments were further conducted for three different reservoir water samples with 30⁻60 ng/L of indulgent 2-MIB. The experimental results demonstrated that preservation with permanganate may have underestimated the 2-MIB concentration in the samples as a result of the formation of manganese dioxide particles in natural water and adsorption of 2-MIB onto the particles. Chlorine was demonstrated to be a good preservation agent for all three tested natural waters since oxidation of 2-MIB was negligible and biodegradation was inhibited. When the residual chlorine concentrations were controlled to be higher than 0.5 mg/L on the final day (day 14) of the experiments, the concentration reduction of 2-MIB became lower than 13% at both of the tested temperatures. The results demonstrated that sodium hypochlorite can be used as an alternative preservation agent for 2-MIB in water before analysis.


Mapping of Current Obstacles for Rationalizing Use of Medicines (CORUM) in Europe: Current Situation and Potential Solutions.
There are increasing concerns regarding the inappropriate use of medicines with expenditure continuing to grow driven by increasing sales in oncology and orphan diseases, enhanced by their emotive nature. As a result, even high income countries are struggling to fund new premium priced medicines. These concerns have resulted in initiatives to better manage the entry of new medicines and enhance the rational use of medicines (RUM). However, there is a need to ascertain the current situation. We sought to address this by developing the Current Obstacles for Rationalizing Use of Medicines in Europe (CORUM) mapping tool to qualitatively investigate the current situation and provide analysis of current views on RUM and interventions among key European payers and their advisers. The findings will be used to provide future guidance. Descriptive study exploring and identifying perceived gaps to achieving optimal RUM. The CORUM tool was based on the WHO 12 key interventions to promote RUM. 62 participants took part with most respondents believing their country could improve RUM capacity. This included educational initiatives on the use of clinical guidelines (90%) and the inclusion of problem-based pharmacotherapy in undergraduate curricula and for Continued Professional Development. Key challenges included a lack of regular updates of guidelines, exacerbated by limited funding and a lack of follow-up to monitor adherence to agreed guidelines. RUM could also be enhanced by the development of regional formularies as well as implementing Drug and Therapeutic Committees where these are currently limited. There also needs to be greater co-ordination between RUM and Health Technology Assessment activities, with countries learning from each other. There is an urgent need to improve RUM through improved educational and other activities among European countries, with countries learning from each other. This will involve addressing current challenges and we will be following this up.


Secondary gastrointestinal cancer in childhood cancer survivors: a cohort study.
Childhood cancer survivors develop gastrointestinal cancer more frequently and at a younger age than the general population, but the risk factors have not been well-characterized. To determine the risk and associated risk factors for gastrointestinal subsequent malignant neoplasms (SMNs) in childhood cancer survivors. Retrospective cohort study. The Childhood Cancer Survivor Study, a multicenter study of childhood cancer survivors diagnosed between 1970 and 1986. 14 358 survivors of cancer diagnosed when they were younger than 21 years of age who survived for 5 or more years after the initial diagnosis. Standardized incidence ratios (SIRs) for gastrointestinal SMNs were calculated by using age-specific population data. Multivariate Cox regression models identified associations between risk factors and gastrointestinal SMN development. At median follow-up of 22.8 years (range, 5.5 to 30.2 years), 45 cases of gastrointestinal cancer were identified. The risk for gastrointestinal SMNs was 4.6-fold higher in childhood cancer survivors than in the general population (95% CI, 3.4 to 6.1). The SIR for colorectal cancer was 4.2 (CI, 2.8 to 6.3). The highest risk for gastrointestinal SMNs was associated with abdominal radiation (SIR, 11.2 [CI, 7.6 to 16.4]). However, survivors not exposed to radiation had a significantly increased risk (SIR, 2.4 [CI, 1.4 to 3.9]). In addition to abdominal radiation, high-dose procarbazine (relative risk, 3.2 [CI, 1.1 to 9.4]) and platinum drugs (relative risk, 7.6 [CI, 2.3 to 25.5]) independently increased the risk for gastrointestinal SMNs. This cohort has not yet attained an age at which risk for gastrointestinal cancer is greatest. Childhood cancer survivors, particularly those exposed to abdominal radiation, are at increased risk for gastrointestinal SMNs. These findings suggest that surveillance of at-risk childhood cancer survivors should begin at a younger age than that recommended for the general population. National Cancer Institute.


HSP70 and HSP90 Differentially Regulate Translocation of Extracellular Antigen to the Cytosol for Cross-Presentation.
Antigens (Ag) from cancer or virus-infected cells must be internalized by dendritic cells (DCs) to be presented to CD8(+) T cells, which eventually differentiate into Ag-specific cytotoxic T lymphocytes (CTLs) that destroy cancer cells and infected cells. This pathway is termed cross-presentation and is also implicated as an essential step in triggering autoimmune diseases such as Type I diabetes. Internalized Ag locates within endosomes, followed by translocation through a putative pore structure spanning endosomal membranes into the cytosol, where it is degraded by the proteasome to generate antigen peptides. During translocation, Ag is believed to be unfolded since the pore size is too narrow to accept native Ag structure. Here, we show that paraformaldehyde-fixed, structurally inflexible Ag is less efficient in cross-presentation because of diminished translocation into the cytosol, supporting the "unfolded Ag" theory. We also show that HSP70 inhibitors block both endogenous and cross-presentation. ImageStream analysis revealed that the inhibition in cross-presentation is not due to blocking of Ag translocation because a HSP70 inhibitor rather facilitates the translocation, which is in marked contrast to the effect of an HSP90 inhibitor that blocks Ag translocation. Our results indicate that Ag translocation to the cytosol in cross-presentation is differentially regulated by HSP70 and HSP90.


Expression of cation exchanger NHE and anion exchanger AE isoforms in primary human bone-derived osteoblasts.
The authors used isoform-specific antibodies against cation (NHE) and anion (AE) exchange isoforms in order to establish their specific expression and localization in dispersed human bone-derived cells. Immunocytochemical preparations of permeabilized osteoblasts probed with polyclonal antibodies were optically analysed by conventional immunofluorescence and con-focal laser scanning microscopy. These techniques demonstrated the abundant presence of epitopes of the cation exchangers NHE1 and NHE3 and the anion exchanger AE2 in these cells. The NHE1 and NHE3 isoform proteins were predominantly located in subplasmalemmal and nucleoplasmic vesicles. The AE2 isoform was densely localized to a subcellular location characteristic of the Golgi complex. The molecular identity of the AE and NHE isoforms was investigated by RT-PCR that confirmed the presence of NHE1 and NHE3 transcripts in addition to NHE4. RT-PCR and diagnostic restriction analysis of amplified AE cDNA established preferential AE2 expression. Since AE2 has been shown to act as a sulfate transporter at low pH, it is possible that it performs this function in the osteoblast Golgi complex where sulfation reactions occur post-translationally on numerous extracellular matrix macromolecules prior to secretion and mineralization. The Na(+)/H(+)exchanger proteins are regulated by mitogenic and non-mitogenic stimuli in the osseus environment and are involved in the large fluxes of ions and protons that necessarily occur during bone formation and resorption and thus play an important role in intracellular ion homeostasis in osteoblasts.


FTO genotype, dietary protein, and change in appetite: the Preventing Overweight Using Novel Dietary Strategies trial.
A common obesity-risk variant rs9939609 in the fat mass- and obesity-associated (FTO) gene was recently shown to affect appetite, and the gene is sensitive to the regulation of amino acids. We examined the interaction between FTO genotype and protein intake on the long-term changes in appetite in a randomized controlled trial. We genotyped FTO rs9939609 in 737 overweight adults in the 2-y Preventing Overweight Using Novel Dietary Strategies trial and assessed 4 appetite-related traits including cravings, fullness, hunger, and prospective consumption. We showed that dietary protein significantly modified genetic effects on changes in food cravings and appetite scores at 6 mo after adjustment for age, sex, ethnicity, baseline body mass index, weight change, and baseline value for respective outcomes (P-interaction = 0.027 and 0.048, respectively). The A allele was associated with a greater decrease in food cravings and appetite scores in participants with high-protein-diet intake (P = 0.027 and 0.047, respectively) but not in subjects in the low-protein-diet group (P = 0.384 and 0.078, respectively). The weight regain from 6 to 24 mo attenuated gene-protein interactions. Protein intakes did not modify FTO genotype effects on other appetite measures. Our data suggest that individuals with the FTO rs9939609 A allele might obtain more benefits in a reduction of food cravings and appetite by choosing a hypocaloric and higher-protein weight-loss diet. This trial was registered at clinicaltrials.gov as NCT00072995.


NPAS1-ARNT and NPAS3-ARNT crystal structures implicate the bHLH-PAS family as multi-ligand binding transcription factors.
The neuronal PAS domain proteins NPAS1 and NPAS3 are members of the basic helix-loop-helix-PER-ARNT-SIM (bHLH-PAS) family, and their genetic deficiencies are linked to a variety of human psychiatric disorders including schizophrenia, autism spectrum disorders and bipolar disease. NPAS1 and NPAS3 must each heterodimerize with the aryl hydrocarbon receptor nuclear translocator (ARNT), to form functional transcription complexes capable of DNA binding and gene regulation. Here we examined the crystal structures of multi-domain NPAS1-ARNT and NPAS3-ARNT-DNA complexes, discovering each to contain four putative ligand-binding pockets. Through expanded architectural comparisons between these complexes and HIF-1α-ARNT, HIF-2α-ARNT and CLOCK-BMAL1, we show the wider mammalian bHLH-PAS family is capable of multi-ligand-binding and presents as an ideal class of transcription factors for direct targeting by small-molecule drugs.


Psychological therapies for chronic pelvic pain: systematic review of randomized controlled trials.
Chronic pelvic pain (CPP), a common cause of disability in women, is a condition best viewed in the biopsychosocial framework. Psychological interventions are frequently considered alongside medical and surgical treatments. Our objective was to evaluate the effectiveness of psychological therapies for the treatment of CPP. Electronic literature searches were conducted in Medline, Embase, PsycInfo and DARE databases from database inception to April 2010. Reference lists of selected articles were searched for further articles. The studies selected were randomized controlled trials of psychological therapies in patients with CPP compared with no treatment, standard gynecological treatment or another form of psychological therapy. Two reviewers independently selected articles without language restrictions and extracted data covering study characteristics, study quality and results. Reduction in pain, measured using visual analog scales or other measurements, was the main outcome measure. Of the 107 citations identified, four studies satisfied the inclusion criteria. Compared with no psychological intervention, therapy produced a standardized mean pain score of -3.27 [95% confidence interval (CI) -4.52 to -2.02] and 1.11 (95% CI -0.05 to 2.27) at 3 months and -3.95 (95% CI -5.35 to -2.55) and 0.54 (95% CI -0.78 to 1.86) at 6 months and greater, based on a visual analog scale score of 0-10. The current evidence does not allow us to conclude whether psychological interventions have an effect on self-reported pain scores in women with CPP.


Mesoporous silica nanoparticles incorporating squaraine-based photosensitizers: a combined experimental and computational approach.
Squaraine dyes, that exhibit intense absorption in the red/near infrared region, have been successfully introduced into mesoporous silica nanoparticles (MSNs) to obtain a nanoplatform for photodynamic therapy. Three brominated squaraine dyes which exhibited good PS performance in solution but a different behaviour in vitro due to cell permeability issues were used as photosensitizers. The effects of the adsorption within the pores of MSNs on the overall UV-Vis-NIR absorption and emission properties as well as on the PS performance was evaluated via a combined experimental and computational physico-chemical approach, which allowed us to correlate the unusual optical properties of two out of three systems with the dimerization of squaraines on the silica surface, with detrimental effects on the PS performance. Conversely, the nanoplatform where the squaraine molecules are adsorbed as monomers on the silica surface exhibited the best activity in singlet oxygen generation.


A new approach to estimating trends in chlamydia incidence.
Directly measuring disease incidence in a population is difficult and not feasible to do routinely. We describe the development and application of a new method for estimating at a population level the number of incident genital chlamydia infections, and the corresponding incidence rates, by age and sex using routine surveillance data. A Bayesian statistical approach was developed to calibrate the parameters of a decision-pathway tree against national data on numbers of notifications and tests conducted (2001-2013). Independent beta probability density functions were adopted for priors on the time-independent parameters; the shapes of these beta parameters were chosen to match prior estimates sourced from peer-reviewed literature or expert opinion. To best facilitate the calibration, multivariate Gaussian priors on (the logistic transforms of) the time-dependent parameters were adopted, using the Matérn covariance function to favour small changes over consecutive years and across adjacent age cohorts. The model outcomes were validated by comparing them with other independent empirical epidemiological measures, that is, prevalence and incidence as reported by other studies. Model-based estimates suggest that the total number of people acquiring chlamydia per year in Australia has increased by ∼120% over 12 years. Nationally, an estimated 356 000 people acquired chlamydia in 2013, which is 4.3 times the number of reported diagnoses. This corresponded to a chlamydia annual incidence estimate of 1.54% in 2013, increased from 0.81% in 2001 (∼90% increase). We developed a statistical method which uses routine surveillance (notifications and testing) data to produce estimates of the extent and trends in chlamydia incidence.


Retrospective review of Japanese sudden unexpected death in infancy: the importance of metabolic autopsy and expanded newborn screening.
Sudden unexpected death in infancy is defined as sudden unexpected death occurring before 12 months of age. The common causes of sudden unexpected death in infancy are infection, cardiovascular anomaly, child abuse, and metabolic disorders. However, the many potential inherited metabolic disorders are difficult to diagnose at autopsy and may therefore be underdiagnosed as a cause of sudden unexpected death in infancy. In the present study we retrospectively reviewed 30 Japanese sudden unexpected death in infancy cases encountered between 2006 and 2009 at our institute. With postmortem blood acylcarnitine analysis and histological examination of the liver, we found two cases of long-chain fatty acid oxidation defects. Molecular analysis revealed that the one patient had a compound heterozygote for a novel mutation (p.L644S) and a disease-causing mutation (p.F383Y) in the carnitine palmitoyltransferase 2 gene. Furthermore, retrospective acylcarnitine analysis of the newborn screening card of this patient was consistent with carnitine palmitoyltransferase II deficiency. Metabolic autopsy and expanded newborn screening would be helpful for forensic scientists and pediatricians to diagnose fatty acid oxidation disorders and prevent sudden unexpected death in infancy.


Comparison of the effects of the uncompetitive N-methyl-D-aspartate antagonist (+-)-5-aminocarbonyl-10,11-dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (ADCI) with its structural analogs dizocilpine (MK-801) and carbamazepine on ethanol withdrawal seizures.
The ability of [(+-)-5-aminocarbonyl-10,11-dihydro-5H-di-benzo [a,d]cyclohepten-5,10-imine (ADCI) and its structural analogs dizocilipine (MK-801) and carbamazepine to block ethanol withdrawal seizures was tested in mice made physically dependent upon ethanol. Three injections of either ADCI (ranging from 1.0-10.0 mg/kg), dizocilpine (ranging from 0.1-1.0 mg/kg) or carbamazepine (ranging from 17-50 mg/kg) were administered during the first 7 hr of ethanol withdrawal. The severity of ethanol withdrawal seizures was rated during the first 11 hr of withdrawal and again at 24 hr after withdrawal of ethanol. ADCI and dizocilpine suppressed the severity and occurrence of the withdrawal seizures in a dose-dependent fashion, whereas carbamazepine was ineffective in blocking the withdrawal seizures. The relative potencies of dizocilpine, ADCI and carbamazepine in suppressing ethanol withdrawal seizures corresponded with the relative potencies of the compounds in displacing [3H]dizocilpine from mouse cortical membrane preparations. These findings are consistent with the suggestion that blockade of N-methyl-D-aspartate-mediated neurotransmission is an effective treatment for decreasing ethanol withdrawal seizures. ADCI also blocked the occurrence of withdrawal-associated whole body tremors, whereas dizocilpine and carbamazepine were ineffective in blocking the tremors. The doses of ADCI, dizocilpine and carbamazepine that resulted in motor incoordination on an accelerating rotarod task were determined in groups of naive mice. Dizocilpine in doses as low as 0.3 mg/kg produced a decreased ability to remain on the rotarod, whereas ADCI up to 30 mg/kg did not affect rotarod performance.(ABSTRACT TRUNCATED AT 250 WORDS)


The meaning and measurement of implementation climate.
Climate has a long history in organizational studies, but few theoretical models integrate the complex effects of climate during innovation implementation. In 1996, a theoretical model was proposed that organizations could develop a positive climate for implementation by making use of various policies and practices that promote organizational members' means, motives, and opportunities for innovation use. The model proposes that implementation climate--or the extent to which organizational members perceive that innovation use is expected, supported, and rewarded--is positively associated with implementation effectiveness. The implementation climate construct holds significant promise for advancing scientific knowledge about the organizational determinants of innovation implementation. However, the construct has not received sufficient scholarly attention, despite numerous citations in the scientific literature. In this article, we clarify the meaning of implementation climate, discuss several measurement issues, and propose guidelines for empirical study. Implementation climate differs from constructs such as organizational climate, culture, or context in two important respects: first, it has a strategic focus (implementation), and second, it is innovation-specific. Measuring implementation climate is challenging because the construct operates at the organizational level, but requires the collection of multi-dimensional perceptual data from many expected innovation users within an organization. In order to avoid problems with construct validity, assessments of within-group agreement of implementation climate measures must be carefully considered. Implementation climate implies a high degree of within-group agreement in climate perceptions. However, researchers might find it useful to distinguish implementation climate level (the average of implementation climate perceptions) from implementation climate strength (the variability of implementation climate perceptions). It is important to recognize that the implementation climate construct applies most readily to innovations that require collective, coordinated behavior change by many organizational members both for successful implementation and for realization of anticipated benefits. For innovations that do not possess these attributes, individual-level theories of behavior change could be more useful in explaining implementation effectiveness. This construct has considerable value in implementation science, however, further debate and development is necessary to refine and distinguish the construct for empirical use.


Analysis of optical elements with the local plane-interface approximation.
The local plane-interface approximation (LPIA) is a method for propagating electromagnetic fields through the inhomogeneous regions (e.g., elements) of an optical system. The LPIA is the superclass of all approximations that replace the usually curved optical interfaces with local tangential planes. Therefore the LPIA is restricted to smooth optical surfaces. A maximum radius of curvature of the optical interface of the order of a few wavelengths is a rough estimate for the validity of the LPIA. Two important approximation levels of the LPIA are the thin-element approximation (TEA) and a geometric-optical version of the LPIA (LPIA(ray)). The latter combines the wave-optical propagation of an electromagnetic field in the homogeneous region of an optical system with a ray-tracing step in the inhomogeneous region. We discuss the regions of validity of the LPIA in general and the approximation levels LPIA(ray) and TEA in detail.


Multiple control elements mediate activation of the murine and human interleukin 12 p40 promoters: evidence of functional synergy between C/EBP and Rel proteins.
Interleukin 12 (IL-12) is a heterodimeric cytokine whose activity is critical for T-helper 1 responses. The gene for the IL-12 p40 subunit is expressed in macrophages following induction by bacterial products, and its expression is augmented by gamma interferon. In this study, we performed a functional analysis of the murine and human p40 promoters in the murine macrophage cell line RAW 264.7. Transcription from the murine p40 promoter was strongly induced by lipopolysaccharide and heat-killed Listeria monocytogenes (HKLM), but promoter activity was not enhanced by gamma interferon. Multiple cis-acting elements involved in activated transcription were identified through an extensive mutant analysis. The most critical element, whose activity is conserved in mice and humans, is located between positions -96 and -88 relative to the murine transcription start site. This element exhibits functional synergy with a previously described NF-kappaB half-site which interacts with Rel proteins. DNase I footprinting and electrophoretic mobility shift assays demonstrated that C/EBP proteins interact with the critical element, but in nuclear extracts, cooperative binding of C/EBP and Rel proteins to their respective sites was not observed. Interestingly, promoter activity was induced by HKLM in the presence of cycloheximide, consistent with induction by posttranslational mechanisms. The results suggest that C/EBP and Rel proteins play important roles in the activation of IL-12 p40 transcription by bacteria. However, many complex interactions will need to be clarified to fully understand p40 regulation.


Engineered living blood vessels: functional endothelia generated from human umbilical cord-derived progenitors.
Tissue-engineered living blood vessels (TEBV) with growth capacity represent a promising new option for the repair of congenital malformations. We investigate the functionality of TEBV with endothelia generated from human umbilical cord blood-derived endothelial progenitor cells. Tissue-engineered living blood vessels were generated from human umbilical cord-derived myofibroblasts seeded on biodegradable vascular scaffolds, followed by endothelialization with differentiated cord blood-derived endothelial progenitor cells. During in vitro maturation the TEBV were exposed to physiologic conditioning in a flow bioreactor. For functional assessment, a subgroup of TEBV was stimulated with tumor necrosis factor-alpha. Control vessels endothelialized with standard vascular endothelial cells were treated in parallel. Analysis of the TEBV included histology, immunohistochemistry, biochemistry (extracellular matrix analysis, DNA), and biomechanical testing. Endothelia were analyzed by flow cytometry and immunohistochemistry (CD31, von Willebrand factor, thrombomodulin, tissue factor, endothelial nitric oxide synthase). Histologically, a three-layered tissue organization of the TEBV analogous to native vessels was observed, and biochemistry revealed the major matrix constituents (collagen, proteoglycans) of blood vessels. Biomechanical properties (Young's modulus, 2.03 +/- 0.65 MPa) showed profiles resembling those of native tissue. Endothelial progenitor cells expressed typical endothelial cell markers CD31, von Willebrand factor, and endothelial nitric oxide synthase comparable to standard vascular endothelial cells. Stimulation with tumor necrosis factor-alpha resulted in physiologic upregulation of tissue factor and downregulation of thrombomodulin expression. These results indicate that TEBV with tissue architecture and functional endothelia similar to native blood vessels can be successfully generated from human umbilical cord progenitor cells. Thus, blood-derived progenitor cells obtained before or at birth may enable the clinical realization of tissue engineering constructs for pediatric applications.


Tuberculosis meningitis, Abbassia Fever Hospital-Naval Medical Research Unit No. 3-Cairo, Egypt, from 1976 to 1996.
A total of 1,430 patients with the presumptive diagnosis of tuberculous meningitis were admitted to the U.S. Naval Medical Research Unit No. 3/Abbassia Fever Hospital in Cairo, Egypt from January 1976 to January 1996. Diagnosis was confirmed by culture of the mycobacteria from the cerebrospinal fluid CSF of 857 patients and these patients are included in the final analysis. There were 497 males and 360 females. The patients ranged in age from five months to 55 years. The number of patients admitted during the months of March, April, and May were more than double those admitted during October, November, and December. The duration of symptoms prior to admission ranged from seven to 90 days (mean = 29.5 days). Upon admission, 4% of the patients were alert, 34% were drowsy, and 62% were in a coma. Of the 857 patients studied, 490 (57%) died, 256 (30%) recovered completely, and 11 (13%) recovered with sequelae. The mortality and neurologic sequelae were directly related to the stage of disease and duration of symptoms prior to admission. Mortality was significantly lower in patients admitted in stage II and or with short duration of disease compared with those in stage III and or with prolonged duration of symptoms prior to admission. The use of dexamethasone in patients with tuberculous meningitis significantly reduced the ocular complications that occur in these patients and also significantly reduced the fatality rate.


Microbial production of bi-functional molecules by diversification of the fatty acid pathway.
Fatty acids that are chemically functionalized at their ω-ends are rare in nature yet offer unique chemical and physical properties with wide ranging industrial applications as feedstocks for bio-based polymers, lubricants and surfactants. Two enzymatic determinants control this ω-group functionality, the availability of an appropriate acyl-CoA substrate for initiating fatty acid biosynthesis, and a fatty acid synthase (FAS) variant that can accommodate that substrate in the initial condensation reaction of the process. In Type II FAS, 3-ketoacyl-ACP synthase III (KASIII) catalyses this initial condensation reaction. We characterized KASIIIs from diverse bacterial sources, and identified variants with novel substrate specificities towards atypical acyl-CoA substrates, including 3-hydroxybutyryl-CoA. Using Alicyclobacillus acidocaldarius KASIII, we demonstrate the in vivo diversion of FAS to produce novel ω-1 hydroxy-branched fatty acids from glucose in two bioengineered microbial hosts. This study unveils the biocatalytic potential of KASIII for synthesizing diverse ω-functionalized fatty acids.


Should global burden of disease estimates include depression as a risk factor for coronary heart disease?
The 2010 Global Burden of Disease Study estimates the premature mortality and disability of all major diseases and injuries. In addition it aims to quantify the risk that diseases and other factors play in the aetiology of disease and injuries. Mental disorders and coronary heart disease are both significant public health issues due to their high prevalence and considerable contribution to global disease burden. For the first time the Global Burden of Disease Study will aim to assess mental disorders as risk factors for coronary heart disease. We show here that current evidence satisfies established criteria for considering depression as an independent risk factor in development of coronary heart disease. A dose response relationship appears to exist and plausible biological pathways have been proposed. However, a number of challenges exist when conducting a rigorous assessment of the literature including heterogeneity issues, definition and measurement of depression and coronary heart disease, publication bias and residual confounding. Therefore, despite some limitations in the available data, it is now appropriate to consider major depression as a risk factor for coronary heart disease in the new Global Burden of Disease Study.


Impairment of TrkB-PSD-95 signaling in Angelman syndrome.
Angelman syndrome (AS) is a neurodevelopment disorder characterized by severe cognitive impairment and a high rate of autism. AS is caused by disrupted neuronal expression of the maternally inherited Ube3A ubiquitin protein ligase, required for the proteasomal degradation of proteins implicated in synaptic plasticity, such as the activity-regulated cytoskeletal-associated protein (Arc/Arg3.1). Mice deficient in maternal Ube3A express elevated levels of Arc in response to synaptic activity, which coincides with severely impaired long-term potentiation (LTP) in the hippocampus and deficits in learning behaviors. In this study, we sought to test whether elevated levels of Arc interfere with brain-derived neurotrophic factor (BDNF) TrkB receptor signaling, which is known to be essential for both the induction and maintenance of LTP. We report that TrkB signaling in the AS mouse is defective, and show that reduction of Arc expression to control levels rescues the signaling deficits. Moreover, the association of the postsynaptic density protein PSD-95 with TrkB is critical for intact BDNF signaling, and elevated levels of Arc were found to impede PSD-95/TrkB association. In Ube3A deficient mice, the BDNF-induced recruitment of PSD-95, as well as PLCγ and Grb2-associated binder 1 (Gab1) with TrkB receptors was attenuated, resulting in reduced activation of PLCγ-α-calcium/calmodulin-dependent protein kinase II (CaMKII) and PI3K-Akt, but leaving the extracellular signal-regulated kinase (Erk) pathway intact. A bridged cyclic peptide (CN2097), shown by nuclear magnetic resonance (NMR) studies to uniquely bind the PDZ1 domain of PSD-95 with high affinity, decreased the interaction of Arc with PSD-95 to restore BDNF-induced TrkB/PSD-95 complex formation, signaling, and facilitate the induction of LTP in AS mice. We propose that the failure of TrkB receptor signaling at synapses in AS is directly linked to elevated levels of Arc associated with PSD-95 and PSD-95 PDZ-ligands may represent a promising approach to reverse cognitive dysfunction.


Common breast cancer susceptibility alleles are associated with tumour subtypes in BRCA1 and BRCA2 mutation carriers: results from the Consortium of Investigators of Modifiers of BRCA1/2.
Previous studies have demonstrated that common breast cancer susceptibility alleles are differentially associated with breast cancer risk for BRCA1 and/or BRCA2 mutation carriers. It is currently unknown how these alleles are associated with different breast cancer subtypes in BRCA1 and BRCA2 mutation carriers defined by estrogen (ER) or progesterone receptor (PR) status of the tumour. We used genotype data on up to 11,421 BRCA1 and 7,080 BRCA2 carriers, of whom 4,310 had been affected with breast cancer and had information on either ER or PR status of the tumour, to assess the associations of 12 loci with breast cancer tumour characteristics. Associations were evaluated using a retrospective cohort approach. The results suggested stronger associations with ER-positive breast cancer than ER-negative for 11 loci in both BRCA1 and BRCA2 carriers. Among BRCA1 carriers, single nucleotide polymorphism (SNP) rs2981582 (FGFR2) exhibited the biggest difference based on ER status (per-allele hazard ratio (HR) for ER-positive = 1.35, 95% CI: 1.17 to 1.56 vs HR = 0.91, 95% CI: 0.85 to 0.98 for ER-negative, P-heterogeneity = 6.5 × 10-6). In contrast, SNP rs2046210 at 6q25.1 near ESR1 was primarily associated with ER-negative breast cancer risk for both BRCA1 and BRCA2 carriers. In BRCA2 carriers, SNPs in FGFR2, TOX3, LSP1, SLC4A7/NEK10, 5p12, 2q35, and 1p11.2 were significantly associated with ER-positive but not ER-negative disease. Similar results were observed when differentiating breast cancer cases by PR status. The associations of the 12 SNPs with risk for BRCA1 and BRCA2 carriers differ by ER-positive or ER-negative breast cancer status. The apparent differences in SNP associations between BRCA1 and BRCA2 carriers, and non-carriers, may be explicable by differences in the prevalence of tumour subtypes. As more risk modifying variants are identified, incorporating these associations into breast cancer subtype-specific risk models may improve clinical management for mutation carriers.


The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin.
The cell substrate attachment (CSAT) antigen is an integral membrane glycoprotein complex that participates in the adhesion of cells to extracellular molecules. The CSAT monoclonal antibody, directed against this complex, inhibited adhesion of cardiac and tendon fibroblasts and skeletal myoblasts to both laminin and fibronectin, thus implicating the CSAT antigen in adhesion to these extracellular molecules. Equilibrium gel filtration was used to explore the hypothesis that the CSAT antigen functions as a cell surface receptor for both laminin and fibronectin. In this technique, designed for rapidly exchanging equilibria, the gel filtration column is pre-equilibrated with extracellular ligand to ensure receptor occupancy during its journey through the column. Both laminin and fibronectin formed complexes with the CSAT antigen. The association with laminin was inhibited by the CSAT monoclonal antibody; the associations with both fibronectin and laminin were inhibited by synthetic peptides containing the fibronectin cell-binding sequence. Estimates of the dissociation constants by equilibrium gel filtration agree well with those available from other measurements. This suggests that these associations are biologically significant. SDS PAGE showed that all three glycoproteins comprising the CSAT antigen were present in the antigen-ligand complexes. Gel filtration and velocity sedimentation were used to show that the three bands comprise and oligomeric complex, which provides an explanation for their functional association. The inhibition of adhesion by the CSAT monoclonal antibody and the association of the purified antigen with extracellular ligands are interpreted as strongly implicating the CSAT antigen as a receptor for both fibronectin and laminin and perhaps for other extracellular molecules as well.


Heat exposure, cardiovascular stress and work productivity in rice harvesters in India: implications for a climate change future.
Excessive workplace heat exposures create well-known risks of heat stroke, and it limits the workers' capacity to sustain physical activity. There is very limited evidence available on how these effects reduce work productivity, while the quantitative relationship between heat and work productivity is an essential basis for climate change impact assessments. We measured hourly heat exposure in rice fields in West Bengal and recorded perceived health problems via interviews of 124 rice harvesters. In a sub-group (n = 48) heart rate was recorded every minute in a standard work situation. Work productivity was recorded as hourly rice bundle collection output. The hourly heat levels (WBGT = Wet Bulb Globe Temperature) were 26-32°C (at air temperatures of 30-38°C), exceeding international standards. Most workers reported exhaustion and pain during work on hot days. Heart rate recovered quickly at low heat, but more slowly at high heat, indicating cardiovascular strain. The hourly number of rice bundles collected was significantly reduced at WBGT>26°C (approximately 5% per°C of increased WBGT). We conclude that high heat exposure in agriculture caused heat strain and reduced work productivity. This reduction will be exacerbated by climate change and may undermine the local economy.


Leaf Spot Disease of Spinach in California Caused by Stemphylium botryosum.
Beginning in 1997, a new disease of spinach was found in the Salinas Valley, Monterey County, CA. Initial symptoms were leaf spots that were 2 to 5 mm in diameter, circular, and gray-green in color. Spots later enlarged, turned tan in color, and became dry and papery in texture, resembling phytotoxicity due to agrichemicals. Although fungal signs were generally absent from the spots, Stemphylium botryosum was consistently isolated and caused identical symptoms when inoculated onto 20 spinach cultivars. Three isolates did not cause disease symptoms when inoculated onto other crop plants representing 16 different genera and a Chenopodium weed species. A fourth isolate showed similar results with the exception of small leaf spots occurring on inoculated fava bean. Isolates produced a Pleospora herbarum teleomorph after 7 months incubation at 5°C. Preliminary experiments with cell-free culture filtrates indicated that phytotoxins apparently were not produced by these isolates. This is the first report of a foliar spinach disease caused by S. botryosum.


G-quadruplexes incorporating modified constituents: a review.
This review summarizes the results of structural studies carried out with analogs of G-quadruplexes built from natural nucleotides. Several dozens of base-, sugar-, and phosphate derivatives of the biological building blocks have been incorporated into more than 50 potentially quadruplex forming DNA and RNA oligonucleotides and the stability and folding topology of the resultant intramolecular, bimolecular and tetramolecular architectures characterized. The TG4T, TG5T, the 15 nucleotide-long thrombin binding aptamer, and the human telomere repeat AG3(TTAG3)3 sequences were modified in most cases, and four guanine analogs can be noted as being particularly useful in structural studies. These are the fluorescent 2-aminopurine, the 8-bromo-, and 8-methylguanines, and the hypoxanthine. The latter three analogs stabilize a given fold in a mixture of structures making possible accurate structural determinations by circular dichroism and nuclear magnetic resonance measurements.


The mouse mammary tumour virus long terminal repeat encodes a type II transmembrane glycoprotein.
Superantigens are products of bacterial or viral origin which stimulate large numbers of T cells as a consequence of the interaction of particular V beta chains of the T cell receptor with class II major histocompatibility complex (MHC) molecules and superantigen on the stimulating cell. The Minor lymphocyte stimulatory (Mls) antigens, originally discovered as strong lymphocyte stimulatory determinants in vitro and subsequently shown to delete T cells expressing specific V beta chains during development, have recently been shown to be genetically linked to endogenous mouse mammary tumour viruses (MTVs). This stimulation is effectuated by an unidentified product encoded by an open reading frame (orf) present in the 3' long terminal repeat (LTR) of MTVs. Using in vitro translation in the presence of rough microsomal vesicles, we show that (i) the orf of MTV encodes a type II transmembrane glycoprotein (N-terminus intracellular, C-terminus extracytoplasmic), and (ii) a cotranslationally secreted orf protein is not produced. We have also isolated and sequenced several endogenous MTV orfs (MTV-1, MTV-6 and MTV-13) which are involved in the deletion of V beta-bearing T cells; each of these sequences are nearly identical to each other. These observations, together with sequence comparisons of several orf genes, lead to a model of action of viral superantigens.


PhenCode: connecting ENCODE data with mutations and phenotype.
PhenCode (Phenotypes for ENCODE; http://www.bx.psu.edu/phencode) is a collaborative, exploratory project to help understand phenotypes of human mutations in the context of sequence and functional data from genome projects. Currently, it connects human phenotype and clinical data in various locus-specific databases (LSDBs) with data on genome sequences, evolutionary history, and function from the ENCODE project and other resources in the UCSC Genome Browser. Initially, we focused on a few selected LSDBs covering genes encoding alpha- and beta-globins (HBA, HBB), phenylalanine hydroxylase (PAH), blood group antigens (various genes), androgen receptor (AR), cystic fibrosis transmembrane conductance regulator (CFTR), and Bruton's tyrosine kinase (BTK), but we plan to include additional loci of clinical importance, ultimately genomewide. We have also imported variant data and associated OMIM links from Swiss-Prot. Users can find interesting mutations in the UCSC Genome Browser (in a new Locus Variants track) and follow links back to the LSDBs for more detailed information. Alternatively, they can start with queries on mutations or phenotypes at an LSDB and then display the results at the Genome Browser to view complementary information such as functional data (e.g., chromatin modifications and protein binding from the ENCODE consortium), evolutionary constraint, regulatory potential, and/or any other tracks they choose. We present several examples illustrating the power of these connections for exploring phenotypes associated with functional elements, and for identifying genomic data that could help to explain clinical phenotypes.


Establishing an evaluation mode with multiple primary outcomes based on combination of diseases and symptoms in TCM clinical trials.
The development of a scientific evaluation tool is a prerequisite to reflect the efficacy of traditional Chinese medicine (TCM). At present, separate analysis of each outcome is the most commonly used method in randomized controlled trials (RCTs) of TCM, while this method has been reported to have some limitations. Combination evaluation of multiple primary outcomes including diseases and symptoms can reflect the efficacy of TCM comprehensively and objectively. Now, several multivariate statistical methods have been proposed to analyze RCTs with multiple clinical outcomes. In this article, we highlighted the two commonly used methods, global statistical test (GST) methodology and alpha-adjustment procedures, and introduced an evaluation mode. First, we used one stage adaptive adjustment strategy to estimate the correlation among multiple primary outcomes in blindness, which can resolve the inappropriate correlation estimation among multiple primary outcomes, then we introduced an adaptation of the Bonferroni procedure which can account for correlated data to calculate the alpha of individual outcome. GST using O'Brien ranking procedure and the corresponding global treatment effect (GTE) measure were used to assess the treatment's global impact. We will conduct a randomized, double-blind, controlled trial with compound Danshen dripping pills for stable angina to explore establishing the evaluation mode. We hope that the introduction of this approach can provide methodological aid for the assessment of therapeutic effect of TCM.


Activity of the farnesyl protein transferase inhibitor SCH66336 against BCR/ABL-induced murine leukemia and primary cells from patients with chronic myeloid leukemia.
BCR/ABL, the oncoprotein responsible for chronic myeloid leukemia (CML), transforms hematopoietic cells through both Ras-dependent and -independent mechanisms. Farnesyl protein transferase inhibitors (FTIs) were designed to block mutant Ras signaling, but they also inhibit the growth of transformed cells with wild-type Ras, implying that other farnesylated targets contribute to FTI action. In the current study, the clinical candidate FTI SCH66336 was characterized for its ability to inhibit BCR/ABL transformation. When tested against BCR/ABL-BaF3 cells, a murine cell line that is leukemogenic in mice, SCH66336 potently inhibited soft agar colony formation, slowed proliferation, and sensitized cells to apoptotic stimuli. Quantification of activated guanosine triphosphate (GTP)-bound Ras protein and electrophoretic mobility shift assays for AP-1 DNA binding showed that Ras effector pathways are inhibited by SCH66336. However, SCH66336 was more inhibitory than dominant-negative Ras in assays of soft agar colony formation and cell proliferation, suggesting activity against targets other than Ras. Cell cycle analysis of BCR/ABL-BaF3 cells treated with SCH66336 revealed G2/M blockade, consistent with recent reports that centromeric proteins that regulate the G2/M checkpoint are critical farnesylated targets of FTI action. Mice injected intravenously with BCR/ABL-BaF3 cells developed acute leukemia and died within 4 weeks with massive splenomegaly, elevated white blood cell counts, and anemia. In contrast, nearly all mice treated with SCH66336 survived and have remained disease-free for more than a year. Furthermore, SCH66336 selectively inhibited the hematopoietic colony formation of primary human CML cells. As an oral, nontoxic compound with a mechanism of action distinct from that of ABL tyrosine kinase inhibition, FTI SCH66336 shows promise for the treatment of BCR/ABL-induced leukemia.


Comparison of endotracheal intubation, laryngeal mask airway, and I-gel in children undergoing strabismus surgery.
Insertion of the advanced airway during induction of general anesthesia can cause undesirable sympathetic stimulation such as increased intraocular pressure (IOP) and hemodynamic parameters. In this study, we compared insertion of three different advanced airway devices; endotracheal tube (ETT), laryngeal mask airway (LMA) and I-gel in terms of IOP, hemodynamic changes and postoperative nausea and vomiting (PONV) following induction of general anesthesia with propofol and remifentanil in children undergoing strabismus surgery. A total of 90 children (5.68 ± 1.49 years old) were randomly assigned to one of the three groups, ETT, LMA, or I-gel insertion as advanced airway devices IOP and also hemodynamic variables were measured before (T0 and T1) and immediately after (T2) the insertion of these airway devices, although 2 min (T3) and 5 min (T4) after it. PONV was assessed about 2 h after the completion of surgery in the recovery room. The mean arterial pressure (MAP), IOP, and systolic and diastolic blood pressures were significantly different between the three groups immediately (T2), 2 min (T3), and 5 min (T4) after the insertion of airway devices. The heart rate (HR) was significantly different between the three groups in all measurement times except of T0. Within-group comparisons showed that the three groups had significant changes in MAP, IOP, HR, systolic and diastolic pressure before and after airway insertion (T1 and T2). The trend in the LMA and ETT groups was descending-ascending-descending, whereas in the I-gel group, it was quite descending. There was no significant difference among the three groups in terms of PONV. As a result, our study showed that, compared with LMA and ETT, the I-gel had less impact on undesirable stress responses and seems to be superior to LMA and ETT in children undergoing strabismus surgery.


Multisite Model of Allosterism for the Adenosine A1 Receptor.
Despite being a target for about one-third of approved drugs, G protein-coupled receptors (GPCRs) still represent a tremendous reservoir for therapeutic strategies against diseases. For example, several cardiovascular and central nervous system conditions could benefit from clinical agents that activate the adenosine 1 receptor (A1R); however, the pursuit of A1R agonists for clinical use is usually impeded by both on- and off-target side effects. One of the possible strategies to overcome this issue is the development of positive allosteric modulators (PAMs) capable of selectively enhancing the effect of a specific receptor subtype and triggering functional selectivity (a phenomenon also referred to as bias). Intriguingly, besides enforcing the effect of agonists upon binding to an allosteric site, most of the A1R PAMs display intrinsic partial agonism and orthosteric competition with antagonists. To rationalize this behavior, we simulated the binding of the prototypical PAMs PD81723 and VCP171, the full-agonist NECA, the antagonist 13B, and the bitopic agonist VCP746. We propose that a single PAM can bind several A1R sites rather than a unique allosteric pocket, reconciling the structure-activity relationship and the mutagenesis results.


[Effect of dongchong xiacao capsule on airway inflammation of asthmatic patients].
To investigate the effect of dongchong xiacao capsule on the airway inflammation of asthmatic patients and to explore the relevant mechanism of therapeutic effect of Dongchongxiacao capsule. Sixty patients with moderate persistent asthma were randomized into the treatment group (n=30) and the control group (n=30). Inhaled corticosteroid and as-needed beta-agonist were used in the treatment group while this therapy plus dongchong xiacao capsule were used in the control group for two months. Serum IL-4, IFN-gamma, sICAM-1, MMP-9, IgG, IgE level were assessed at randomization and 2 months after randomization. The serum level of IgE, sICAM-1, IL-4 and MMP-9 of the treatment group was lowered to a greater degree than that of the control group (P < 0.05 or P < 0.01 ). Dongchong xiacao capsule can reduce the serum markers of airway inflammation, which suggests this therapy bares the anti-inflammation effects probably through regulating the balance of TH1/TH2, inhibiting the activity of adherence molecule and reducing IgE production. It may also have the effect of reversing airway remodeling, which needs further research to determine.


Scene-based nonuniformity corrections for optical and SWIR pushbroom sensors.
We propose and evaluate several scene-based methods for computing nonuniformity corrections for visible or near-infrared pushbroom sensors. These methods can be used to compute new nonuniformity correction values or to repair or refine existing radiometric calibrations. For a given data set, the preferred method depends on the quality of the data, the type of scenes being imaged, and the existence and quality of a laboratory calibration. We demonstrate our methods with data from several different sensor systems and provide a generalized approach to be taken for any new data set.


CiiiDER: A tool for predicting and analysing transcription factor binding sites.
The availability of large amounts of high-throughput genomic, transcriptomic and epigenomic data has provided opportunity to understand regulation of the cellular transcriptome with an unprecedented level of detail. As a result, research has advanced from identifying gene expression patterns associated with particular conditions to elucidating signalling pathways that regulate expression. There are over 1,000 transcription factors (TFs) in vertebrates that play a role in this regulation. Determining which of these are likely to be controlling a set of genes can be assisted by computational prediction, utilising experimentally verified binding site motifs. Here we present CiiiDER, an integrated computational toolkit for transcription factor binding analysis, written in the Java programming language, to make it independent of computer operating system. It is operated through an intuitive graphical user interface with interactive, high-quality visual outputs, making it accessible to all researchers. CiiiDER predicts transcription factor binding sites (TFBSs) across regulatory regions of interest, such as promoters and enhancers derived from any species. It can perform an enrichment analysis to identify TFs that are significantly over- or under-represented in comparison to a bespoke background set and thereby elucidate pathways regulating sets of genes of pathophysiological importance.


A Foilized Polyethylene Pouch for the Prevention of Transmission of HIV from Mother to Child.
Many children become HIV+ due to mother-to-child transmission, a risk that can be largely eliminated if infants ingest antiretroviral (ARV) medications immediately after birth. As most mothers in Africa deliver at home, the ARV must be provided at their last antenatal visit, sometimes months before birth. No current drug delivery system allows the mother to store the medication at home long enough to be effective. We propose a preserving, foilized, polyethylene pouch to be pre-dosed and sealed by a pharmacist for later delivery to the newborn.Pouches were filled with 0.6 ml of Nevirapine (NVP). Thirty-three pouches were immediately studied to measure the impact of medication handling (oxygen, light, filling and sealing the pouches). The remaining samples were stored for up to one year at three storage conditions (25°C/60%RH, 30°C/65%RH, and 40°C/75%RH). Every two months, moisture loss, preservative concentration, impurity concentrations and NVP concentration were measured. Flora and fauna challenges were conducted.THE POUCH NEARLY ELIMINATED MOISTURE LOSS: pouches lost less than 0.7% of their weight over twelve months. As expected, exposing the medication to light, oxygen, and handling significantly affected the sacrificial preservative concentrations (Propyl paraben dropped 38%, Methyl paraben dropped 12% at time point zero). However, after the initial time point, preservative levels were stable in the package over twelve months under all storage conditions (4.1% average concentration drop), leaving sufficient preservatives to protect the medication. The concentration of NVP changed an average of only 1.3% over all storage conditions and times points (maximum 1.4%).We conclude that the foilized polyethylene pouch can preserve NVP, and perhaps other ARV's, for up to one year.


Health-related needs of people with multiple chronic diseases: differences and underlying factors.
To examine the health-related needs of people with multiple chronic diseases in the Netherlands compared to people with one chronic disease, and to identify different subgroups of multimorbid patients based on differences in their health problems. Participants were 1092 people with one or more chronic diseases of a nationwide prospective panel study on the consequences of chronic illness in the Netherlands. They completed the EQ-6D, a multi-dimensional questionnaire on health problems (October 2013). Chi-square tests and analyses of variance were performed to test for differences between multimorbid patients and patients with one chronic disease. To identify subgroups of multimorbid patients, cluster analysis was performed and differences in EQ-6D scores between clusters were tested with Chi-square tests. Multimorbid patients (51 % of the total sample) experience more problems in most health domains than patients with one chronic disease. Almost half (44 %) of the multimorbid people had many health problems in different domains. These people were more often female, had a smaller household size, had a lower health literacy, and suffered from more chronic diseases. Remarkably, a small subgroup of multimorbid patients (4 %, mostly elderly males) is characterized by all having cognitive problems. Based on the problems they experience, we conclude that patients with multimorbidity have relatively many and diverse health-related needs. Extensive health-related needs among people with multimorbidity may relate not only to the number of chronic diseases they suffer from, but also to their patient characteristics. This should be taken into account, when identifying target groups for comprehensive support programmes.


Reduced Serum Levels of Cluster of Differentiation 200 in Alcohol-Dependent Patients.
Chronic alcohol consumption can activate and dysregulate the neuroimmune system which leads to neuroinflammation. Neuroimmune regulatory proteins (NIReg) (e.g. Cluster of Differentiation 200 (CD200)) are the regulators of innate immune response and are responsible for silencing the innate immunity and suppression of inflammation. In this study, we explored the changes of serum levels of CD200 in patients with alcohol dependence at baseline, after one-week alcohol withdrawal and after one-month of alcohol abstinence. Seventeen patients with alcohol dependence admitted for de-addiction treatment and 12 healthy controls were enrolled in the study. Blood samples were collected at baseline, after one-week, and after one-month, and CD200 levels were measured using enzyme-linked immunosorbent assay kit and compared with the healthy controls. The serum level of the neuroimmune regulatory protein CD200 in alcohol dependent group (at baseline) was significantly lower compared to healthy controls (p=0.003), and increased after one-week, and one-month period. The present study indicates that decrease of CD200 serum levels in alcohol dependent patients and its rise during alcohol withdrawal and abstinence may provide a preliminary evidence of the role of neuroimmune regulatory proteins in neuroadaptation during alcohol withdrawal.


Ion gradients in xylem exudate and guttation fluid related to tissue ion levels along primary leaves of barley.
The concentration of ions in plant cells and tissues is an essential factor in determining physiological function. In the present study, we established that concentration gradients of mobile ions exist in both xylem exudates and tissues within a barley (Hordeum vulgare) primary leaf. For K(+) and NO3 (-) , ion concentrations generally decreased from the leaf base to the tip in both xylem exudates and tissues. Ion gradients were also found for Pi and Cl(-) in the xylem. The hydathode strongly absorbed Pi and re-translocated it to the rest of the plant, whereas Cl(-) was extruded. The ion concentration gradients developed early during leaf growth, increased as the tissue aged and remained under both high and low transpiration conditions. Measurement of the expression profiles of Pi, K(+) and NO3 (-) transporters along the longitudinal axis of the leaf revealed that some transporters are more expressed at the hydathode, but for most transporters, there was no significant variation along the leaf. The mechanisms by which longitudinal ion gradients develop in leaves and their physiological functions are discussed.


Compact efficient passively Q-switched Nd:GdVO4/PPLN/Cr4+:YAG tunable intracavity optical parametric oscillator.
We report on a compact efficient diode pumped passively Q-switched Nd:GdVO4/Cr4+:YAG/PPLN intracavity optical parametric oscillator (OPO) with a shared-resonator configuration. Experimental results reveal that the amplitude stability of the shared-resonator configuration is substantially superior to that of the conventional coupledresonator configuration. At a diode pump power of 15 W, the compact intracavity OPO cavity produces the average power greater than 900 mW with a pulse repetition rate of 36 kHz. The output pulses noticeably display the mode-locking phenomenon that leads to the maximum peak power to be higher than 20 kW.


Video-assisted thoracoscopic surgery for pulmonary aspergilloma.
Surgical management of pulmonary aspergilloma in symptomatic patients offers a significant chance of cure. Video-assisted thoracic surgery is a valid alternative for properly selected cases. We herein report our experience with thoracoscopic management of pulmonary aspergilloma. This retrospective analysis was performed on 41 patients operated between 2012 to 2015. The patient records were thoroughly analyzed for demography, clinical presentation, computed tomography , the procedure performed , post-operative complications and course during 6 month's follow up. Out of total 41 patients, 23 (56%) were treated by VATS and 18 (44%) by thoracotomy. Average intraoperative blood loss was 214 ml (±106) in VATS group and 461 ml (±167) in thoracotomy. Mean operative time was 162 (±14) minutes in VATS and 239 (±12) minutes in thoracotomy group. In VATS group, postoperative complications were found in 5 patients and in 11 patients in the thoracotomy group. Average duration of chest tube was 5.43 () days in VATS group and 8.94 () days in thoracotomy group. Average length of hospital stay was 5.04 in VATS group and 6.55 days in thoracotomy group. VATS for pulmonary aspergilloma, if applicable, may be a safe and efficacious option in experienced hands. Simple aspergilloma, in particular , is considered to be a good indication for VATS. Some cases of complex aspergilloma may also be amenable to VATS. However, the long term results need to be further analyzed using a larger study group.