Fortification of dark chocolate with spray dried black mulberry (Morus nigra) waste extract encapsulated in chitosan-coated liposomes and bioaccessability studies.
Fine-disperse anionic liposomes containing black mulberry (Morus nigra) extract (BME) were prepared by high pressure homogenization at 25,000 psi. Primary liposomes were coated with cationic chitosan (0.4, w/v%) using the layer-by-layer depositing method and mixed with maltodextrin (MD) (20, w/v%) prior to spray drying. After that, spray dried liposomal powders containing BME were added to chocolates with alkalization degrees (pH 4.5, 6, 7.5) at conching temperatures of 40 °C, 60 °C, and 80 °C. The results showed that, compared to spray dried extract, chitosan coated liposomal powders provided better protection of anthocyanin content in both increased temperature and pH. In addition, encapsulation in liposomes enhanced in vitro bioaccessability of anthocyanins. Chocolate was fortified with encapsulated anthocyanins maximum 76.8% depending on conching temperature and pH.


Microsurgical, endoscopic, and shunt management of pediatric temporosylvian arachnoid cysts: a comparative study.
Arachnoid cysts (ACs) are most frequently located in the middle cranial fossa. Some patients are asymptomatic whereas others exhibit signs of increased intracranial pressure, seizures, or cognitive and behavioral symptoms. When ACs do require treatment, the optimal surgical technique remains controversial. This study was conducted to assess the most effective surgical treatment for these cysts. The authors retrospectively reviewed 240 temporal intracranial ACs managed over a 25-year period in their pediatric neurosurgical unit. Pre- and posttreatment results were clinically and radiologically assessed. A majority of male patients (74.6%) with an overall median age of 6.9 years were included. The mean cyst size was 107 cm3; the Galassi classification showed 99 (41.3%) type I, 77 (32.1%) type II, and 64 (26.7%) type III cysts. Forty-four ACs (18.3%) were diagnosed after rupture. Surgical management was performed by microsurgery (28.3%), endoscopic cyst fenestration (14.6%), cystoperitoneal shunting (CPS; 16.2%), or subdural shunting (10%). Furthermore, 74 children (30.8%) did not undergo operations. After a mean follow-up of 4.1 years, the mean percentage decrease in cyst volume and the overall rate of clinical improvement did not significantly differ. The endoscopy group had earlier complications and a shorter event-free survival (EFS) time (EFS at 3 years = 67.7%, vs 71.5% and 90.5% for CPS and microsurgery, respectively; p < 0.007) and presented with more subdural hematomas compared to the microsurgery group (p < 0.005). The microsurgery group also showed a tendency for longer cystocisternostomy permeability than the endoscopy group. Concerning the management of unruptured symptomatic temporal ACs, microsurgery appears to be the most effective treatment, with longer EFS and fewer complications compared to shunting or endoscopy.


Food quality in producer-grazer models: a generalized analysis.
Stoichiometric constraints play a role in the dynamics of natural populations but are not explicitly considered in most mathematical models. Recent theoretical works suggest that these constraints can have a significant impact and should not be neglected. However, it is not yet resolved how stoichiometry should be integrated in population dynamical models, as different modeling approaches are found to yield qualitatively different results. Here we investigate a unifying framework that reveals the differences and commonalities between previously proposed models for producer-grazer systems. Our analysis reveals that stoichiometric constraints affect the dynamics mainly by increasing the intraspecific competition between producers and by introducing a variable biomass conversion efficiency. The intraspecific competition has a strongly stabilizing effect on the system, whereas the variable conversion efficiency resulting from a variable food quality is the main determinant for the nature of the instability once destabilization occurs. Only if the food quality is high can an oscillatory instability, as in the classical paradox of enrichment, occur. While the generalized model reveals that the generic insights remain valid in a large class of models, we show that other details such as the specific sequence of bifurcations encountered in enrichment scenarios can depend sensitively on assumptions made in modeling stoichiometric constraints.


Effect of donor recipient age match on survival after first deceased donor renal transplantation.
Donor-recipient age matching has been proposed as a means of improving overall outcomes in deceased donor renal transplantation. It was hypothesized that donor-recipient age matching would improve patient survival time in younger recipients while not adversely affecting patient survivals in older recipients because they seldom outlive their grafts. By use of data from United Network of Organ Sharing Standard Transplant and Analysis and Research Files 50,320 patients were identified who underwent a first deceased donor renal transplantation between January 1, 1990, and December 31, 1997. Adjusted patient survival and death-with-graft function patient survival were analyzed from the date of transplantation. Patient survival was affected by donor age for all recipient age groups, including recipients older than 55 yr. The effect of donor age on patient survival is greater than that seen with HLA matching. The effect of donor age on patient survival persisted even when censoring recipients in whom grafts failed before death, suggesting that both longevity and quality of graft function are important in patient survival. Donor-recipient age matching is occurring to a limited degree in this population. Donor-recipient age matching would improve survival in younger recipients but would adversely affect survival in older patients by reducing the availability of younger donor kidneys for this group. The issue of donor-recipient age matching needs to be debated among the public and transplantation community so that a logical and just system can be developed.


Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning.
We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies.


Genome-scale reconstruction of Gcn4/ATF4 networks driving a growth program.
Growth and starvation are considered opposite ends of a spectrum. To sustain growth, cells use coordinated gene expression programs and manage biomolecule supply in order to match the demands of metabolism and translation. Global growth programs complement increased ribosomal biogenesis with sufficient carbon metabolism, amino acid and nucleotide biosynthesis. How these resources are collectively managed is a fundamental question. The role of the Gcn4/ATF4 transcription factor has been best studied in contexts where cells encounter amino acid starvation. However, high Gcn4 activity has been observed in contexts of rapid cell proliferation, and the roles of Gcn4 in such growth contexts are unclear. Here, using a methionine-induced growth program in yeast, we show that Gcn4/ATF4 is the fulcrum that maintains metabolic supply in order to sustain translation outputs. By integrating matched transcriptome and ChIP-Seq analysis, we decipher genome-wide direct and indirect roles for Gcn4 in this growth program. Genes that enable metabolic precursor biosynthesis indispensably require Gcn4; contrastingly ribosomal genes are partly repressed by Gcn4. Gcn4 directly binds promoter-regions and transcribes a subset of metabolic genes, particularly driving lysine and arginine biosynthesis. Gcn4 also globally represses lysine and arginine enriched transcripts, which include genes encoding the translation machinery. The Gcn4 dependent lysine and arginine supply thereby maintains the synthesis of the translation machinery. This is required to maintain translation capacity. Gcn4 consequently enables metabolic-precursor supply to bolster protein synthesis, and drive a growth program. Thus, we illustrate how growth and starvation outcomes are both controlled using the same Gcn4 transcriptional outputs that function in distinct contexts.


The cumulative effect of core lifestyle behaviours on the prevalence of hypertension and dyslipidemia.
Most cardiovascular disease (CVD) occurs in the presence of traditional risk factors, including hypertension and dyslipidemia, and these in turn are influenced by behavioural factors such as diet and lifestyle. Previous research has identified a group at low risk of CVD based on a cluster of inter-related factors: body mass index (BMI) < 25 Kg/m2, moderate exercise, alcohol intake, non-smoking and a favourable dietary pattern. The objective of this study was to determine whether these factors are associated with a reduced prevalence of hypertension and dyslipidemia in an Irish adult population. The study was a cross-sectional survey of 1018 men and women sampled from 17 general practices. Participants completed health, lifestyle and food frequency questionnaires and provided fasting blood samples for analysis of glucose and insulin. We defined a low risk group based on the following protective factors: BMI <25 kg/m2; waist-hip ratio (WHR) <0.85 for women and <0.90 for men; never smoking status; participants with medium to high levels of physical activity; light alcohol consumption (3.5-7 units of alcohol/week) and a "prudent" diet. Dietary patterns were assessed by cluster analysis. We found strong significant inverse associations between the number of protective factors and systolic blood pressure, diastolic blood pressure and dyslipidemia. The prevalence odds ratio of hypertension in persons with 1, 2, 3, > or = 4 protective factors relative to those with none, were 1.0, 0.76, 0.68 and 0.34 (trend p < 0.01). The prevalence odds ratio of dyslipidemia in persons with 1, 2, 3, > or = 4 protective factors relative to those with none were 0.83, 0.98, 0.49 and 0.24 (trend p = 0.001). Our findings of a strong inverse association between low risk behaviours and two of the traditional risk factors for CVD highlight the importance of 'the causes of the causes' and the potential for behaviour modification in CVD prevention at a population level.


Possible variation of the human oral bacterial community after wearing removable partial dentures by DGGE.
Although it is well-known that variations of the microbial community in a specific location of human body may be associated with some diseases, the developing change of the oral microbiota related to oral diseases before and after wearing the removable partial dentures (RPD) is not completely understood. In this study, three kinds of samples (saliva, supra- and subgingival plaque, and oral mucosal surfaces) were collected from the 10-patients group at three different times: before, 1-month and 6-months after the treatment. Ten healthy adults were also selected as the control group. Denaturing gradient gel electrophoresis was applied to identify the bacterial profiles and to analyze the dynamics of the oral microbial population in the pre- and post-therapy. The ANOVA of Repeated Measurement Data indicated that, in the saliva and mucosal surfaces, wearing RPDs caused significant change of numbers of amplicons. As many as 607 amplicons were chosen to cut out and re-amplify by PCR. After cloning and sequencing, a total of 16 bacterial genera were identified. The health-associated genera such as Streptococcus, Neisseria, Rothia, Corynebacterium, Leptotrichia, Gemella, Veillonella, Selenomona and Actinomyces tended to decrease, whereas the disease-associated species including Streptococcus mutans tended to increase. In general, wearing RPDs influenced the diversity of the bacterial species in the oral microbial ecosystem. It is noteworthy that the oral environment will be changed from the healthy status towards the disease status after the treatment.


Acetyl-l-carnitine deficiency in patients with major depressive disorder.
The lack of biomarkers to identify target populations greatly limits the promise of precision medicine for major depressive disorder (MDD), a primary cause of ill health and disability. The endogenously produced molecule acetyl-l-carnitine (LAC) is critical for hippocampal function and several behavioral domains. In rodents with depressive-like traits, LAC levels are markedly decreased and signal abnormal hippocampal glutamatergic function and dendritic plasticity. LAC supplementation induces rapid and lasting antidepressant-like effects via epigenetic mechanisms of histone acetylation. This mechanistic model led us to evaluate LAC levels in humans. We found that LAC levels, and not those of free carnitine, were decreased in patients with MDD compared with age- and sex-matched healthy controls in two independent study centers. Secondary exploratory analyses showed that the degree of LAC deficiency reflected both the severity and age of onset of MDD. Moreover, these analyses showed that the decrease in LAC was larger in patients with a history of treatment-resistant depression (TRD), among whom childhood trauma and, specifically, a history of emotional neglect and being female, predicted the decreased LAC. These findings suggest that LAC may serve as a candidate biomarker to help diagnose a clinical endophenotype of MDD characterized by decreased LAC, greater severity, and earlier onset as well as a history of childhood trauma in patients with TRD. Together with studies in rodents, these translational findings support further exploration of LAC as a therapeutic target that may help to define individualized treatments in biologically based depression subtype consistent with the spirit of precision medicine.


Establishment of an internationally agreed minimum data set for acute telestroke.
Globally, the use of telestroke programmes for acute care is expanding. Currently, a standardised set of variables for enabling reliable international comparisons of telestroke programmes does not exist. The aim of the study was to establish a consensus-based, minimum dataset for acute telestroke to enable the reliable comparison of programmes, clinical management and patient outcomes. An initial scoping review of variables was conducted, supplemented by reaching out to colleagues leading some of these programmes in different countries. An international expert panel of clinicians, researchers and managers (n = 20) from the Australasia Pacific region, USA, UK and Europe was convened. A modified-Delphi technique was used to achieve consensus via online questionnaires, teleconferences and email. Overall, 533 variables were initially identified and harmonised into 159 variables for the expert panel to review. The final dataset included 110 variables covering three themes (service configuration, consultations, patient information) and 12 categories: (1) details about telestroke network/programme (n = 12), (2) details about initiating hospital (n = 10), (3) telestroke consultation (n = 17), (4) patient characteristics (n = 7), (5) presentation to hospital (n = 5), (6) general clinical care within first 24 hours (n = 10), (7) thrombolysis treatment (n = 10), (8) endovascular treatment (n = 13), (9) neurosurgery treatment (n = 8), (10) processes of care beyond 24 hours (n = 7), (11) discharge information (n = 5), (12) post-discharge and follow-up data (n = 6). The acute telestroke minimum dataset provides a recommended set of variables to systematically evaluate acute telestroke programmes in different countries. Adoption is recommended for new and existing services.


Methotrexate dose-escalation studies in transgenic mice and marrow transplant recipients expressing drug-resistant dihydrofolate reductase activity.
Methotrexate (MTX) dose-escalation studies were conducted in two inbred lines of FVB/N transgenic mice expressing distinct drug-resistant dihydrofolate reductases (DHFRs) and in animals transplanted with transgenic marrow. Survival of animals expressing a tryptophan-31 variant DHFR transgene was only slightly improved over that of normal animals, and survival of tryptophan-31 variant DHFR marrow transplant recipients was indistinguishable from that of normal animals (at a MTX dose of 4 mg/kg i.p. daily). In contrast, extended survival was observed for animals expressing an arginine-22 variant (Arg22) DHFR transgene, with the last three of eight animals in this group succumbing at a final MTX dose of 14 mg/kg i.p. daily. Survival was slightly reduced for normal animals transplanted with Arg22 marrow. Interestingly, demise of animals in both Arg22 groups was not associated with the profound drop in hematocrit levels usually observed in MTX-treated animals. These animals were instead characterized by severe atrophy of the gastrointestinal tract, whereas hematocrit levels and marrow histology were relatively normal. Kidney pathology (mesangiocapillary glomerulopathy) was also observed in Arg22 marrow recipients but not in Arg22 transgenics, consistent with expression of the drug-resistance gene in kidney tissues of the transgenics, as demonstrated by ribonuclease protection analysis. Immediate dose-response studies in Arg22 marrow transplant recipients defined a maximum tolerated dose of 4 mg/kg/day MTX, 2 to 3 times that of animals transplanted with normal marrow or of normal untransplanted animals. These results define the extent of chemoprotection afforded by drug-resistant DHFR expression and serve to identify alternate sites of toxicity in animals administered the higher levels of MTX afforded by drug-resistant DHFR expression in the marrow.


Surface Modified Biodegradable Electrospun Membranes as a Carrier for Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells.
Human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells are currently undergoing clinical trials to treat retinal degenerative diseases. Transplantation of hESC-RPE cells in conjuction with a supportive biomaterial carrier holds great potential as a future treatment for retinal degeneration. However, there has been no such biodegradable material that could support the growth and maturation of hESC-RPE cells so far. The primary aim of this work was to create a thin porous poly (L-lactide-co-caprolactone) (PLCL) membrane that could promote attachment, proliferation, and maturation of the hESC-RPE cells in serum-free culture conditions. The PLCL membranes were modified by atmospheric pressure plasma processing and coated with collagen IV to enhance cell growth and maturation. Permeability of the membranes was analyzed with an Ussing chamber system. Analysis with scanning electron microscopy, contact angle measurement, atomic force microscopy, and X-ray photoelectron spectroscopy demonstrated that plasma surface treatment augments the surface properties of the membrane, which enhances the binding and conformation of the protein. Cell proliferation assays, reverse transcription-polymerase chain reaction, indirect immunofluoresence staining, trans-epithelial electrical resistance measurements, and in vitro phagocytosis assay clearly demonstrated that the plasma treated PLCL membranes supported the adherence, proliferation, maturation and functionality of hESC-RPE cells in serum-free culture conditions. Here, we report for the first time, how PLCL membranes can be modified with atmospheric pressure plasma processing to enable the formation of a functional hESC-RPE monolayer on a porous biodegradable substrate, which have a potential as a tissue-engineered construct for regenerative retinal repair applications.


Dynamic distribution patterns of ribosomal DNA and chromosomal evolution in Paphiopedilum, a lady's slipper orchid.
Paphiopedilum is a horticulturally and ecologically important genus of ca. 80 species of lady's slipper orchids native to Southeast Asia. These plants have long been of interest regarding their chromosomal evolution, which involves a progressive aneuploid series based on either fission or fusion of centromeres. Chromosome number is positively correlated with genome size, so rearrangement processes must include either insertion or deletion of DNA segments. We have conducted Fluorescence In Situ Hybridization (FISH) studies using 5S and 25S ribosomal DNA (rDNA) probes to survey for rearrangements, duplications, and phylogenetically-correlated variation within Paphiopedilum. We further studied sequence variation of the non-transcribed spacers of 5S rDNA (5S-NTS) to examine their complex duplication history, including the possibility that concerted evolutionary forces may homogenize diversity. 5S and 25S rDNA loci among Paphiopedilum species, representing all key phylogenetic lineages, exhibit a considerable diversity that correlates well with recognized evolutionary groups. 25S rDNA signals range from 2 (representing 1 locus) to 9, the latter representing hemizygosity. 5S loci display extensive structural variation, and show from 2 specific signals to many, both major and minor and highly dispersed. The dispersed signals mainly occur at centromeric and subtelomeric positions, which are hotspots for chromosomal breakpoints. Phylogenetic analysis of cloned 5S rDNA non-transcribed spacer (5S-NTS) sequences showed evidence for both ancient and recent post-speciation duplication events, as well as interlocus and intralocus diversity. Paphiopedilum species display many chromosomal rearrangements--for example, duplications, translocations, and inversions--but only weak concerted evolutionary forces among highly duplicated 5S arrays, which suggests that double-strand break repair processes are dynamic and ongoing. These results make the genus a model system for the study of complex chromosomal evolution in plants.


Common variants in mismatch repair genes associated with increased risk of sperm DNA damage and male infertility.
The mismatch repair (MMR) pathway plays an important role in the maintenance of the genome integrity, meiotic recombination and gametogenesis. This study investigated whether genetic variations in MMR genes are associated with an increased risk of sperm DNA damage and male infertility. We selected and genotyped 21 tagging single nucleotide polymorphisms (SNPs) in five MMR genes (MLH1, MLH3, PMS2, MSH4 and MSH5) using the SNPstream 12-plex platform in a case-control study of 1,292 idiopathic infertility patients and 480 fertile controls in a Chinese population. Sperm DNA damage levels were detected with the Tdt-mediated dUTP nick end labelling (TUNEL) assay in 450 cases. Fluorescence resonance energy transfer (FRET) and co-immunoprecipitation techniques were employed to determine the effects of functional variants. One intronic SNP in MLH1 (rs4647269) and two non-synonymous SNPs in PMS2 (rs1059060, Ser775Asn) and MSH5 (rs2075789, Pro29Ser) seem to be risk factors for the development of azoospermia or oligozoospermia. Meanwhile, we also identified a possible contribution of PMS2 rs1059060 to the risk of male infertility with normal sperm count. Among patients with normal sperm count, MLH1 rs4647269 and PMS2 rs1059060 were associated with increased sperm DNA damage. Functional analysis revealed that the PMS2 rs1059060 can affect the interactions between MLH1 and PMS2. Our results provide evidence supporting the involvement of genetic polymorphisms in MMR genes in the aetiology of male infertility.


The epidemiology of migraine.
This article provides a review of the epidemiology and risk factors for migraine in population studies, as well as patterns for healthcare use. The burden and costs of migraine, as well as risk factors for disease progression, are also discussed. Although migraine is a remarkably common cause of temporary disability, many persons with migraine, even those with disabling headache, have never consulted a physician for the problem. Prevalence is highest in women, in persons between the ages of 25 and 55 years, and, at least in the United States, in individuals from lower income households. However, prevalence is high in groups other than these high-risk groups. In a subgroup of patients, migraine may be a progressive disorder.


GPs' job satisfaction: doctors who chose general practice early or late.
In the UK many practising GPs did not choose general practice as their first choice of career when they originally graduated as doctors. To compare job satisfaction of GPs who chose general practice early or later in their career. Questionnaires were sent to all UK-trained doctors who graduated in selected years between 1993 and 2000. Questionnaires were sent to the doctors 1, 3, 7 and 10 years after graduation. Of all 3082 responders working in general practice in years 7 and 10, 38% had first specified general practice as their preferred career when responding 1 year after graduation, 19% by year 3, 21% by year 5, and 22% after year 5. Job satisfaction was high and, generally, there was little difference between the first three groups (although, when different, the most positive responses were from the earliest choosers); but there were slightly lower levels of job satisfaction in the 'more than 5 years' group. For example, in response to the statement 'I find enjoyment in my current post', the percentages agreeing in the four groups, respectively, were 91.5%, 91.1%, 91.0% and 88.2%. In response to 'I am doing interesting and challenging work' the respective percentages were 90.2%, 88.0%, 86.6% and 82.6%. Job satisfaction levels were generally high among the late choosers as well as the early choosers. On this evidence, most doctors who turn to general practice, after preferring another specialty in their early career, are likely to have a satisfying career.


Analysis of Immunologic Function Changes in Lichen Planus After Clinical Treatment.
BACKGROUND Lichen planus (LP) is a common chronic superficial skin lesion that causes chronic or sub-acute inflammatory disorders. LP can affect the oral cavity, skin, mucous membrane, and other body parts, and features include repeat attacks and long duration, leading to lower quality of life. This study aimed to analyze the changes of immunologic function before and after treatment of LP. MATERIAL AND METHODS Thirty cutaneous LP patients were selected. Peripheral blood was collected in the morning before and after treatment. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient method. Flow cytometry was used to detect T cell subpopulation CD4⁺ T cells and CD8⁺ T to calculate CD4⁺ T/CD8⁺ T ratio. Enzyme-linked immunosorbent assay (ELISA) was adopted to detect the helper T-cell (Th) factor IL-2, IFN-γ, IL-4, IL-6, IL-17, and IL-22 levels. RESULTS Compared with before treatment, the expressions of CD4⁺ T cells and CD8⁺ T cells were decreased, while the proportion of CD4⁺ T/CD8⁺ T were significantly elevated after treatment. IL-2 and IFN-γ secretion were markedly increased, whereas IL-4, IL-6, IL-17, and IL-22 were significantly reduced after treatment (P<0.05). CONCLUSIONS LP treatment reduces the distribution of CD4⁺ T cells and CD8⁺ T cells, and promotes the changes of Th1, Th2, and Th17 cytokines secretion.


Characterization of in planta-induced rust genes isolated from a haustorium-specific cDNA library.
Rust fungi are plant parasites that depend on living host tissue for growth. For invasion of leaves, dikaryotic urediospores differentiate germ tubes and infection structures that penetrate through stomata. Biotrophic growth occurs by intercellular mycelia that form haustoria within host cells. A cDNA library was constructed from haustoria isolated from broad bean leaves infected by Uromyces fabae. Differential screening revealed that a high proportion (19%) of the haustorial cDNAs are specifically expressed in planta but are not expressed, or are much weaker, in germlings or infection structures produced in vitro. A total of 31 different in planta-induced genes (PIGs) were identified. Some of the PIGs are highly expressed in haustoria. The PIGs are single or low copy number genes in the rust genome. A variety of developmentally regulated expression patterns of PIG mRNAs were observed. Sequence analysis of PIG cDNAs revealed similarities to genes encoding proteins involved in amino acid transport, thiamine biosynthesis, short-chain dehydrogenases, metallothioneins, cytochrome P-450 monooxygenases, and peptidyl-prolyl isomerases.


Repairs of the rotator cuff. Correlation of functional results with integrity of the cuff.
We evaluated the results of 105 operative repairs of tears of the rotator cuff of the shoulder in eighty-nine patients at an average of five years postoperatively. We correlated the functional result with the integrity of the cuff, as determined by ultrasonography. Eighty per cent of the repairs of a tear involving only the supraspinatus tendon were intact at the time of the most recent follow-up, while more than 50 per cent of the repairs of a tear involving more than the supraspinatus tendon had a recurrent defect. Older patients and patients in whom a larger tear had been repaired had a greater prevalence of recurrent defects. At the time of the most recent follow-up, most of the patients were more comfortable and were satisfied with the result of the repair, even when they had sonographic evidence of a recurrent defect. The shoulders in which the repaired cuff was intact at the time of follow-up had better function during activities of daily living and a better range of active flexion (129 +/- 20 degrees compared with 71 +/- 41 degrees) compared with the shoulders that had a large recurrent defect. Similar correlations were noted for the range of active external and internal rotation and for strength of flexion, abduction, and internal rotation. In the shoulders in which the cuff was not intact, the degree of functional loss was related to the size of the recurrent defect.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of carbon monoxide donor CORM-2 on vitamin D3 metabolism.
Carbon monoxide (CO) interferes with cytochrome-dependent cellular functions and acts as gaseous transmitter. CO is released from CO-releasing molecules (CORM) including tricarbonyl-dichlororuthenium (II) dimer (CORM-2), molecules considered for the treatment of several disorders including vascular dysfunction, inflammation, tissue ischemia and organ rejection. Cytochrome P450-sensitive function include formation of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) by renal 25-hydroxyvitamin D3 1-alpha-hydroxylase (Cyp27b1). The enzyme is regulated by PTH, FGF23 and klotho. 1,25(OH)2D3 regulates Ca(2+) and phosphate transport as well as klotho expression. The present study explored, whether CORM-2 influences 1,25(OH)2D3 formation and klotho expression. Mice were treated with intravenous CORM-2 (20 mg/kg body weight). Plasma 1,25(OH)2D3 and FGF23 concentrations were determined by ELISA, phosphate, calcium and creatinine concentrations by colorimetric methods, transcript levels by quantitative RT-PCR and protein expression by western blotting. Fgf23 mRNA transcript levels were further determined in rat osteosarcoma UMR106 cells without or with prior treatment for 24 hours with 20 µM CORM-2. CORM-2 injection within 24 hours significantly increased FGF23 plasma levels and decreased 1,25(OH)2D3 plasma levels, renal Cyp27b1 gene expression as well as renal klotho protein abundance and transcript levels. Moreover, treatment of UMR106 cells with CORM-2 significantly increased Fgf23 transcript levels. CO-releasing molecule CORM-2 enhances FGF23 expression and release and decreases klotho expression and 1,25(OH)2D3 synthesis. © 2013 S. Karger AG, Basel.


Worldwide trends in incidence in occupational allergy and asthma.
Recent improvements in the methods for analyzing trends in occupational health surveillance and the prospect of future improvements in the collecting and sharing of electronic data alongside increasing availability of linked datasets make this a good time to review the existing literature on trends in occupational allergy and asthma (OAA). There is a notable lack of reports of recent trends in OAA in the academic literature and much of the published work comes from European countries. The incidence of OAA appears to be declining based on physician-reporting or recognized compensation claims for the countries with published data. However, we need to be cautious in interpreting this as a decline in the 'true' incidence of OAA. Few of the studies adjusted appropriately for changes in the population at risk and one of the most robust study designs showed no change in the incidence of allergic contact dermatitis in contrast to the other studies. Many existing datasets have the potential to be used to examine trends, and studies from Canada show the potential of using linked databases for surveillance. We hope that this review will encourage improvements in the analysis, and more dissemination, of trends.


Nucleotide sequence of the chloramphenicol resistance determinant of the streptococcal plasmid pIP501.
We have sequenced the chloramphenicol resistance determinant (cat) of plasmid pIP501 from Streptococcus agalactiae to investigate its relationship with other cognate cat determinants. Sequence analysis revealed that it exhibits a high degree of similarity with the cat genes of plasmids pC221 and pUB112 from Staphylococcus aureus and pSCS1 from Staphylococcus intermedius. These genes, however, display several differences in their regulatory and coding regions. These results demonstrate that the cat determinant of plasmid pIP501 belongs to the pC221 subgroup of CAT variants.


NUT midline carcinoma: an aggressive intrathoracic neoplasm.
Nuclear protein in testis (NUT) midline carcinoma (NMC) is a poorly differentiated squamous cell carcinoma that is characterized by a balanced translocation between chromosomes 15 and 19 [t(15;19)(q14;p13.1)]. This genetic aberration results in the fusion of the NUT gene on chromosome 15 to the bromodomain containing 4 (BRD4) gene on chromosome 19. The resultant BRD4-NUT fusion oncogene leads to global hypoacetylation and transcriptional repression of genes required for differentiation." Although it was first reported in 1991 by Kubonishi et al., awareness of this condition remains low and the diagnosis is overlooked initially in a number of patients. A 36-year-old man complained of cough and right-sided chest pain for 3 weeks before presentation. Imaging studies revealed a right hilar mass, and a bronchoscopic biopsy was consistent with an aggressive poorly differentiated neoplasm. A combination of cisplatin, ifosfamide, and etoposide was administered for two cycles without any improvement. A repeat core biopsy showed focal squamous differentiation; and given the clinical presentation along with the histologic features, NMC was considered in the differential diagnosis. Immunohistochemical staining for NUT was positive, and dual-color break-apart fluorescence in situ hybridization demonstrated BRD4-NUT rearrangement, thereby confirming a diagnosis of NMC. Our patient was subsequently enrolled on a phase 1 clinical trial of a novel, orally bioavailable bromodomain and extra terminal inhibitor, GSK525762 (NCT01587703). This report illustrates the challenges in diagnosing this rare malignancy, and highlights new treatment options for these patients.


An electron-beam CT approach for transvenous coronary arteriography.
The goal is to image the 3D anatomy of the major coronary arteries within a single breath-hold and using a single intravenous injection of contrast agent. With use of an electron-beam CT (EBCT) scanner, operated in its multislice scan mode (using all four target rings, each scanning two nominal 8 mm thick slices in a total time interval of 224 ms), a radiologically realistic thorax phantom containing a heart with coronary arteries (opacified with 40 mg iodine/ml) was scanned six times with the patient table advanced by a 2 mm step into the scanner between each scan. The normalized and scaled sinogram data were transferred to an off-line computer and reconstructed using our own implementation of a 3D algebraic reconstruction technique that takes into account the exact 3D relationship of the X-ray source and detector elements of the EBCT. A single volume image consisting of 40 slices, at 2 mm intervals and each uniformly 3.7 mm thick, depicted the lumina of the coronary arteries with greater anatomic detail than the "original" images at 8 mm, primarily because of the reduced partial volume effect. Our preliminary results suggest that EBCT, operated in multislice scan mode, may be useful for transvenous coronary angiography performed within a single breath-hold.


Energy balance and body composition during US Army special forces training.
Small Unit Tactics (SUT) is a 64-day phase of the Special Forces Qualification Course designed to simulate real-world combat operations. Assessing the metabolic and physiological responses of such intense training allows greater insights into nutritional requirements of soldiers during combat. The purpose of this study was to examine energy balance around specific training events, as well as changes in body mass and composition. Data were collected from 4 groups of soldiers (n = 36) across 10-day periods. Participants were 28 ± 5 years old, 177 ± 6 cm tall, and weighed 83 ± 7 kg. Doubly labeled water (D2(18)O) was used to assess energy expenditure. Energy intake was calculated by subtracting energy in uneaten foods from known energy in distributed foods in individually packaged combat rations or in the dining facility. Body composition was estimated from skinfold thickness measurements on days 0 and 64 of the course. Simulated urban combat elicited that largest energy deficit (11.3 ± 2.3 MJ·day(-1) (2700 ± 550 kcal·day(-1)); p < 0.05), and reduction in body mass (3.3 ± 1.9 kg; p < 0.05), during SUT, while energy balance was maintained during weapons familiarization training and platoon size raids. Over the entire course body mass decreased by 4.2 ± 3.7 kg (p < 0.01), with fat mass decreasing by 2.8 ± 2.0 kg (p < 0.01) and fat-free mass decreasing by 1.4 ± 2.8 kg (p < 0.05). The overall reduction in body mass suggests that soldiers were in a negative energy balance during SUT, with high energy deficit being observed during strenuous field training.


Biased towards food: Electrophysiological evidence for biased attention to food stimuli.
We investigated the neural mechanisms involved in bias for food stimuli in our visual environment using event related lateralized (ERL) responses. The participants were presented with a cue (food or non-food item) to either identify or hold in working memory. Subsequently, they had to search for a target in a 2-item display where target and distractor stimuli were each flanked by a picture of a food or a non-food item. The behavioural data showed that performance was strongly affected by food cues, especially when food was held in WM compared to when the cues were merely identified. The temporal dynamics of electrophysiological measures of attention (the N1pc and N2pc) showed that the orienting of attention towards food stimuli was associated with two different mechanisms; an early stage of attentional suppression followed by a later stage of attentional orienting towards food stimuli. In contrast, non-food cues were associated only with the guidance of attention to or away from cued stimuli on valid and invalid trials. The results demonstrate that food items, perhaps due to their motivational significance modulate the early orienting of attention, including an initial suppressive response to food items.


Genetic diversity of expressed Plasmodium falciparum var genes from Tanzanian children with severe malaria.
Severe malaria has been attributed to the expression of a restricted subset of the var multi-gene family, which encodes for Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1). PfEMP1 mediates cytoadherence and sequestration of infected erythrocytes into the post-capillary venules of vital organs such as the brain, lung or placenta. var genes are highly diverse and can be classified in three major groups (ups A, B and C) and two intermediate groups (B/A and B/C) based on the genomic location, gene orientation and upstream sequences. The genetic diversity of expressed var genes in relation to severity of disease in Tanzanian children was analysed. Children with defined severe (SM) and asymptomatic malaria (AM) were recruited. Full-length var mRNA was isolated and reversed transcribed into var cDNA. Subsequently, the DBL and N-terminal domains, and up-stream sequences were PCR amplified, cloned and sequenced. Sequences derived from SM and AM isolates were compared and analysed. The analysis confirmed that the var family is highly diverse in natural Plasmodium falciparum populations. Sequence diversity of amplified var DBL-1α and upstream regions showed minimal overlap among isolates, implying that the var gene repertoire is vast and most probably indefinite in endemic areas. var DBL-1α sequences from AM isolates were more diverse with more singletons found (p<0.05) than those from SM infections. Furthermore, few var DBL-1α sequences from SM patients were rare and restricted suggesting that certain PfEMP1 variants might induce severe disease. The genetic sequence diversity of var genes of P. falciparum isolates from Tanzanian children is large and its relationship to disease severity has been studied. Observed differences suggest that different var genes might have fundamentally different roles in the host-parasite interaction. Further research is required to examine clear disease-associations of var gene subsets in different geographical settings. The importance of very strict clinical definitions and appropriate large control groups needs to be emphasized for future studies on disease associations of PfEMP1.


Genetic polymorphisms in organic cation transporter 1 (OCT1) in Chinese and Japanese populations exhibit altered function.
Organic cation transporter 1 (OCT1; SLC22A1) seems to play a role in the efficacy and disposition of the widely used antidiabetic drug metformin. Genetic variants in OCT1 have been identified largely in European populations. Metformin is increasingly being used in Asian populations where the incidence of type 2 diabetes (T2D) is on the rise. The goal of this study is to identify genetic variants of OCT1 in Chinese and Japanese populations, which may potentially modulate response to metformin. We used recent data from the 1000 Genomes Project (Chinese and Japanese) and direct sequencing of selected amplicons of OCT1 in 66 DNA samples from Japanese patients with T2D. A total of six nonsynonymous variants were identified. Three of them (Q97K, P117L, and R206C) had not been functionally characterized previously and had allele frequencies of 0.017, 0.023 and 0.008, respectively. The uptake of metformin in cells expressing Q97K, P117L, and R206C was significantly reduced relative to the OCT1 reference (62 ± 4.3, 55 ± 6.8, and 22 ± 1.5% for Q97K, P117L, and R206C, respectively). Kinetic studies indicated that P117L and R206C exhibited a reduced V(max), whereas Q97K showed an increased K(m). The green fluorescent protein (GFP)-tagged Q97K and P117L variants localized to the plasma membrane, whereas the GFP-tagged R206C was retained mainly in the endoplasmic reticulum. Replacement of the highly conserved R206 with different amino acids modulated the subcellular localization and function of the transporter. This study suggests that nonsynonymous variants of OCT1 in Chinese and Japanese populations may affect the differential response to metformin.


Multi-Source Transfer Learning via Ensemble Approach for Initial Diagnosis of Alzheimer's Disease.
Alzheimer's disease (AD) is one of the most common progressive neurodegenerative diseases, and the number of AD patients has increased year after year with the global aging trend. The onset of AD has a long preclinical stage. If doctors can make an initial diagnosis in the mild cognitive impairment (MCI) stage, it is possible to identify and screen those at a high-risk of developing full-blown AD, and thus the number of new AD patients can be reduced. However, there are problems with the medical datasets including AD data, such as insufficient number of samples and different data distributions. Transfer learning, which can effectively solve the problem of distribution discrepancy between training and test data and an insufficient number of target samples, has attracted increasing attention over recent years. In this paper, we propose a multi-source ensemble transfer learning (METL) approach by introducing ensemble learning and our tri-transfer model that uses Tri-Training, which ensures the transferability of source data by the tri-transfer model and high performance through ensemble learning. The experimental results on the benchmark and AD datasets demonstrate that our proposed approach has effective transferability, robustness, and feasibility, and is superior to existing algorithms. Based on METL, we propose an auxiliary diagnosis system for the initial diagnosis of AD, which helps doctors identify patients in the MCI stage as quickly as possible and with high accuracy so that measures can be taken to prevent or delay the occurrence of AD.


Viscoelasticity of dense suspensions of thermosensitive microgel mixtures undergoing colloidal gelation.
Dense suspensions of temperature (T)-sensitive poly(N-isopropyl acrylamide) (N) and poly(N-isopropyl methacrylamide) (NM) microgel mixtures with different volume transition temperatures (T and T, respectively; T < T) exhibit a characteristic T-dependent viscoelasticity due to T-induced changes in the type of interparticle interaction as well as the volume fraction of each gel. In the range of T < T, where the swollen microgels with repulsive interparticle interactions are densely packed, the equilibrium modulus (G) decreases upon heating due entirely to the packing effect, i.e., a reduction in the total volume fraction of the microgels (φ). At T > T where the attractive interparticle interactions between dehydrated and hydrophobic microgels emerge, the suspensions show solid-like elastic properties due to the network-like flocculation of the shrunken microgels (colloidal gelation), even when φ becomes considerably lower than the threshold for randomly close packing. The T-dependence of G shows a minimum at a characteristic temperature (TB; TB > T) due to the competition between the repulsive interparticle interactions from the packing effect and electrostatic force, and the attractive interactions from the hydrophobicity. The TB in N/NM mixture suspensions shifts to a higher value with a decrease in N content in the mixtures (XN), accompanied by a discontinuous-like change at a specific value of XN (XN*). The TB at every value of XN agrees approximately with the temperature where the total volume fraction of the attractive hydrophobic microgels is 0.3 regardless of microgel type (N or NM). The discontinuous-like variation in TB at XN* reflects the change in the network-like flocculation particles, from only attractive N microgels in the high XN regime, to the attractive N and NM microgel mixtures in the moderate XN regime. The requirement of the repulsive electrostatic force with an appropriate strength for the stability of the network-like flocculation is also demonstrated using the PNIPAM-co-fumaric acid (NF) microgel suspensions at various pH.


Inner privacy of conscious experiences and quantum information.
The human mind is constituted by inner, subjective, private, first-person conscious experiences that cannot be measured with physical devices or observed from an external, objective, public, third-person perspective. The qualitative, phenomenal nature of conscious experiences also cannot be communicated to others in the form of a message composed of classical bits of information. Because in a classical world everything physical is observable and communicable, it is a daunting task to explain how an empirically unobservable, incommunicable consciousness could have any physical substrates such as neurons composed of biochemical molecules, water, and electrolytes. The challenges encountered by classical physics are exemplified by a number of thought experiments including the inverted qualia argument, the private language argument, the beetle in the box argument and the knowledge argument. These thought experiments, however, do not imply that our consciousness is nonphysical and our introspective conscious testimonies are untrustworthy. The principles of classical physics have been superseded by modern quantum physics, which contains two fundamentally different kinds of physical objects: unobservable quantum state vectors, which define what physically exists, and quantum operators (observables), which define what can physically be observed. Identifying consciousness with the unobservable quantum information contained by quantum physical brain states allows for application of quantum information theorems to resolve possible paradoxes created by the inner privacy of conscious experiences, and explains how the observable brain is constructed by accessible bits of classical information that are bound by Holevo's theorem and extracted from the physically existing quantum brain upon measurement with physical devices.


Methylation of a histone mimic within the histone methyltransferase G9a regulates protein complex assembly.
Epigenetic gene silencing in eukaryotes is regulated in part by lysine methylation of the core histone proteins. While histone lysine methylation is known to control gene expression through the recruitment of modification-specific effector proteins, it remains unknown whether nonhistone chromatin proteins are targets for similar modification-recognition systems. Here we show that the histone H3 methyltransferase G9a contains a conserved methylation motif with marked sequence similarity to H3 itself. As with methylation of H3 lysine 9, autocatalytic G9a methylation is necessary and sufficient to mediate in vivo interaction with the epigenetic regulator heterochromatin protein 1 (HP1), and this methyl-dependent interaction can be reversed by adjacent G9a phosphorylation. NMR analysis indicates that the HP1 chromodomain recognizes methyl-G9a through a binding mode similar to that used in recognition of methyl-H3K9, demonstrating that the chromodomain functions as a generalized methyl-lysine binding module. These data reveal histone-like modification cassettes - or "histone mimics" - as a distinct class of nonhistone methylation targets and directly extend the principles of the histone code to the regulation of nonhistone proteins.


Electrophysiological characterization of harmane-induced activation of mesolimbic dopamine neurons.
It has been suggested that the beta-carbolines harmane and norharmane may be involved in the pathophysiology of Parkinson's disease, psychosis and addiction, but the mechanisms of these possible effects remain to be elucidated. In the present study, the effects of the two compounds were examined by using in vivo extracellular recordings of ventral tegmental dopamine neurons. The effects of harmane (2mg/kg) and norharmane (2mg/kg), were compared to those of nicotine (11microg/kg), of cotinine (0.5mg/kg), of the monoamine-oxidase-A inhibitor befloxatone (0.12mg/kg), and of the monoamine-oxidase-B inhibitor selegiline (0.5mg/kg). The effects of harmane were also tested after pre-treatment with the nicotine receptor antagonist mecamylamine. The results show that all substances, except befloxatone, activate the firing and/or burst activity of dopamine neurons. The increase in firing rate produced by harmane was approximately 18 times greater than that produced by nicotine. Such powerful excitation of dopamine neurons by harmane may in part explain its involvement in neurotoxicity, psychosis and addiction. The absence of effect of befloxatone supports the hypothesis that the effect of harmane is not related to its monoamine-oxidase-A inhibitory properties. Mecamylamine inhibited by approximately 80% the activity of harmane, indicating that the activating effect of harmane on dopamine neurons involves several mechanisms, among which activation of nicotinic receptors likely has a prominent importance. The results of the present study support the hypothesis that harmane could be a tobacco (or smoke) component other than nicotine involved in tobacco dependence.


Computational approaches to protein inference in shotgun proteomics.
Shotgun proteomics has recently emerged as a powerful approach to characterizing proteomes in biological samples. Its overall objective is to identify the form and quantity of each protein in a high-throughput manner by coupling liquid chromatography with tandem mass spectrometry. As a consequence of its high throughput nature, shotgun proteomics faces challenges with respect to the analysis and interpretation of experimental data. Among such challenges, the identification of proteins present in a sample has been recognized as an important computational task. This task generally consists of (1) assigning experimental tandem mass spectra to peptides derived from a protein database, and (2) mapping assigned peptides to proteins and quantifying the confidence of identified proteins. Protein identification is fundamentally a statistical inference problem with a number of methods proposed to address its challenges. In this review we categorize current approaches into rule-based, combinatorial optimization and probabilistic inference techniques, and present them using integer programming and Bayesian inference frameworks. We also discuss the main challenges of protein identification and propose potential solutions with the goal of spurring innovative research in this area.


Mapping the digital food environment: a scoping review protocol.
Food environments are the interface through which people interact with the broader food system. They are a key determinant of healthy and sustainable diets. The widespread use of digital technology in late modernity and the shift towards a digital society have posed new challenges for nutrition and health, with a concomitant surge in research on social media, digital health promotion interventions, and more recently, increasing interest in digital food marketing. While the literature is abundant on studies linking food, nutrition and digital technology, the effort to conceptualise and describe the digital food environment is new. This scoping review aims to support the development of a definition of the digital food environment and characterise it, along with key thematic research trends on this topic and potential consequences for nutrition and health. The planned scoping review will be supported by the methodological framework proposed by Arksey and O'Malley and further developed by Levac et al. Development and reporting will follow the Preferred Reporting Items for Systematic Reviews and MetaAnalyses-Extension for Scoping Reviews (PRISMA-ScR) checklist and guidelines. The development of the search strategy was guided by the food environment conceptual framework developed by Turner et al. Four databases will be searched: MEDLINE, EMBASE, Scopus and Web of Science. Citation searching will be applied to identify additional studies, through checking of reference lists of primary studies and reviews. Studies in English, published from the year 2000 onwards, will be included. No geographical or population limits will be applied. Data will be extracted and analysed using a standardised charting tool. No ethical approval is required for this study. The results will be submitted to an international peer-reviewed journal and scientific conferences. They will be disseminated through digital science communication platforms, including academic social media, to amplify its reach and usefulness.


Trends in adolescent pregnancies at Tygerberg Hospital, 1976-1980.
This study was undertaken to highlight trends relating to pregnancies among females aged less than 21 years seen at Tygerberg Hospital, Parowvallei, CP, during the 5-year period 1976-1980 inclusive. There was a fairly steady decline in the pregnancy rate for Coloured adolescents over this period. The rate for Black adolescents increased after a 3-year decline, and that for Whites also increased after a 2-year drop. The caesarean section rate increased after a marked drop in 1978, both in White adolescents and in the White group as a whole, possibly due to an increase in the instrumental delivery rate in 1978. The caesarean section rate has increased steadily among non-Whites, while the forceps delivery rate for non-Whites declined to below the normal level of 15% and that for Whites fell after a steep increase in 1978. Vacuum extractions appeared to be very popular during the first 3 years, but the rates declined rapidly towards 1979-1980, particularly in the White group. Pre-eclamptic toxaemia remains a problem among White adolescents. Among both Whites and non-Whites 1978 seems to have been a disastrous year as far as the outcome of pregnancies is concerned, with an increase in the incidence of miscarriage. A drop in the non-White stillbirth rate in 1978 was followed by a sharp rise in 1979-1980. The incidence of abruptio placentae and placenta praevia rose in 1978, but in 1979-1980 dropped to below 10% for Whites and to below 15% for non-Whites. Premature deliveries are still common among non-Whites, with average incidences of 23% for the 5-year period for non-Whites and 8,5% for Whites. Pregnancy rates for unmarried Coloured and Black adolescents remain high, and the rate for Whites increased slightly over the 5-year period.


Endoscopic Severity Predicts Long-Term Prognosis in Crohn's Disease Patients with Clinical Remission.
Mucosal healing has emerged as a desirable treatment goal in clinical practice for patients with Crohn's disease (CD). The aim of this study was to assess the relationship between endoscopic activity and the long-term prognosis of CD using simple endoscopic score for Crohn's disease (SESCD) and Rutgeerts' score. We conducted a cohort study in clinical practice at a single center. Among CD patients who underwent colonoscopy between July 2008 and June 2011 at our hospital, 131 patients with clinical remission were selected, and the patients were divided into 2 groups: a non-surgical group (n = 84) and a surgical group (n = 47). The primary endpoint of this study was to assess the associations between variables and clinical relapse after endoscopic procedures. The cut-off levels of SESCD or Rutgeerts' score for the prediction of relapse were also assessed in patients with clinical remission. In the non-surgical group, SESCD and C-reactive protein at baseline were significantly higher in patients who had clinical recurrence than in patients who maintained remission. A factor of SESCD ≤2 was independently associated with sustained remission, even in patients with clinical remission. In the surgical group, patients with Rutgeerts' scores ≤1 had significantly prolonged clinical remission compared to patients with Rutgeerts' scores ≥3. A cut-off value of SESCD ≤2 and a Rutgeerts' score ≤1 enabled the prediction of long-term prognosis. These cut-off values could be used in clinical trials of endoscopic remission from the point of view of the clinical outcomes of CD.


Effectiveness of Tai Chi on Physical and Psychological Health of College Students: Results of a Randomized Controlled Trial.
To investigate the effectiveness and safety of Tai Chi Chuan (TCC) on physical and psychological health of college students. Two hundred six college students were recruited and randomly allocated to a control group or a TCC exercise group in an equal ratio. Participants in the control group were instructed to maintain their original activity level and those in the TCC exercise group received 12 weeks of TCC exercise training based on their original activity level. Physical and psychological outcomes were evaluated at baseline, 13 weeks and 25 weeks. Intention-to-treat analysis was performed for the above outcomes. Compared with the control group, the TCC exercise group showed significant improvements at the end of the 12-week intervention period for flexibility (length of Sit and Reach (cm): TCC group 14.09±7.40 versus control 12.88±6.57, P = 0.039 adjusted for its baseline measures using a general linear model) and balance ability (open eyes perimeter: TCC group 235.6(191~314) versus control 261(216~300); closed eyes perimeter: TCC group 370.5 (284~454) versus control 367 (293~483); P = 0.0414, 0.008, respectively, adjusted for corresponding baseline measures using a general linear model). No significant changes in other physical and mental outcomes were found between the two groups. No adverse events were reported during the study period. TCC exercise was beneficial in college students for improving flexibility and balance capability to some extent, compared with usual exercise. Chinese Clinical Trial Registry ChiCTR-TRC-13003328.


Longitudinal Cognitive Trajectories of Women Veterans from the Women's Health Initiative Memory Study.
A comparison of longitudinal global cognitive functioning in women Veteran and non-Veteran participants in the Women's Health Initiative (WHI). We studied 7,330 women aged 65-79 at baseline who participated in the WHI Hormone Therapy Trial and its ancillary Memory Study (WHIMS). Global cognitive functioning (Modified Mini-Mental State Examination [3MSE]) in Veterans (n = 279) and non-Veterans (n = 7,051) was compared at baseline and annually for 8 years using generalized linear modeling methods. Compared with non-Veterans, Veteran women were older, more likely to be Caucasian, unmarried, and had higher rates of educational and occupational attainment. Results of unadjusted baseline analyses suggest 3MSE scores were similar between groups. Longitudinal analyses, adjusted for age, education, ethnicity, and WHI trial assignment revealed differences in the rate of cognitive decline between groups over time, such that scores decreased more in Veterans relative to non-Veterans. This relative difference was more pronounced among Veterans who were older, had higher educational/occupational attainment and greater baseline prevalence of cardiovascular risk factors (e.g., smoking) and cardiovascular disease (e.g., angina, stroke). Veteran status was associated with higher prevalence of protective factors that may have helped initially preserve cognitive functioning. However, findings ultimately revealed more pronounced cognitive decline among Veteran relative to non-Veteran participants, likely suggesting the presence of risks that may impact neuropathology and the effects of which were initially masked by Veterans' greater cognitive reserve.


Causal implication by rhythmic transcranial magnetic stimulation of alpha frequency in feature-based local vs. global attention.
Although oscillatory activity in the alpha band was traditionally associated with lack of alertness, more recent work has linked it to specific cognitive functions, including visual attention. The emerging method of rhythmic transcranial magnetic stimulation (TMS) allows causal interventional tests for the online impact on performance of TMS administered in short bursts at a particular frequency. TMS bursts at 10 Hz have recently been shown to have an impact on spatial visual attention, but any role in featural attention remains unclear. Here we used rhythmic TMS at 10 Hz to assess the impact on attending to global or local components of a hierarchical Navon-like stimulus (D. Navon (1977) Forest before trees: The precedence of global features in visual perception. Cognit. Psychol., 9, 353), in a paradigm recently used with TMS at other frequencies (V. Romei, J. Driver, P.G. Schyns & G. Thut. (2011) Rhythmic TMS over parietal cortex links distinct brain frequencies to global versus local visual processing. Curr. Biol., 2, 334-337). In separate groups, left or right posterior parietal sites were stimulated at 10 Hz just before presentation of the hierarchical stimulus. Participants had to identify either the local or global component in separate blocks. Right parietal 10 Hz stimulation (vs. sham) significantly impaired global processing without affecting local processing, while left parietal 10 Hz stimulation vs. sham impaired local processing with a minor trend to enhance global processing. These 10 Hz outcomes differed significantly from stimulation at other frequencies (i.e. 5 or 20 Hz) over the same site in other recent work with the same paradigm. These dissociations confirm differential roles of the two hemispheres in local vs. global processing, and reveal a frequency-specific role for stimulation in the alpha band for regulating feature-based visual attention.


Influence of activation origin, lead number, and lead configuration on the noninvasive electrophysiologic detection of cardiac allograft rejection.
An improved method of cardiac allograft surveillance, less invasive than endomyocardial biopsy (EMB), is required as the scope of cardiac transplantation increases. The correlation between changes in telemetrically recorded intramyocardial electrograms and the degree of rejection was examined. The electrophysiological parameter assessed was the unipolar peak-to-peak amplitude (UPPA), defined as the magnitude of the first, fast negative deflection of the QRS complex in unipolar electrograms. A canine model of intrathoracic heterotopic cardiac transplantation was employed in which each graft was instrumented with four unipolar intramyocardial electrodes (two left ventricle [LV] and two right ventricle [RV]) connected to two telemetric pacemakers. Immunosuppression was begun at operation and continued for a variable period of time following the acquisition of control data (postoperative days 6-10). Graft status was monitored via biweekly EMB. Intramyocardial electrograms were recorded from each lead during sinus rhythm and 36 atrial and ventricular pacing protocols on a daily basis. The daily UPPA was calculated for each rhythm sequence and expressed as percent of control UPPA. EMBs were qualitatively graded and assigned a quantitative rejection score (0 = none; 3 = severe). Criteria for rejection were rejection score (RS) greater than or equal to 0.66 or daily UPPA less than 85% control. Eight animals were included for study and, in all animals, rejection was documented. A total of 30 EMBs were obtained; in 11 EMBs rejection was present by histological criteria. Analysis of grouped UPPA data from all four leads revealed a sensitivity and specificity of 91% and 84%, respectively, in the detection of rejection. Analysis of grouped sinus UPPA data from the two LV leads revealed a sensitivity of 94% and 91%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Impact of micronutrient fortification of yoghurt on micronutrient status markers and growth - a randomized double blind controlled trial among school children in Bangladesh.
Interventions providing foods fortified with multiple micronutrients can be a cost-effective and sustainable strategy to improve micronutrient status and physical growth of school children. We evaluated the effect of micronutrient-fortified yoghurt on the biochemical status of important micronutrients (iron, zinc, iodine, vitamin A) as well as growth indicators among school children in Bogra district of Bangladesh. In a double-masked randomized controlled trial (RCT) conducted in 4 primary schools, 1010 children from classes 1-4 (age 6-9 years) were randomly allocated to receive either micronutrient fortified yoghurt (FY, n = 501) or non-fortified yoghurt (NFY, n = 509). For one year, children were fed with 60 g yoghurt everyday providing 30% RDA for iron, zinc, iodine and vitamin A. Anthropometric measurements and blood/urine samples were collected at base-, mid- and end-line. All children (FY, n = 278, NFY, n = 293) consenting for the end-line blood sample were included in the present analyses. Both groups were comparable at baseline for socio-economic status variables, micronutrient status markers and anthropometry measures. Compliance was similar in both the groups. At baseline 53.4% of the population was anemic; 2.1% was iron deficient (ferritin <15.0 μg/L and TfR > 8.3 mg/L). Children in the FY group showed improvement in Hb (mean difference: 1.5; 95% CI: 0.4-2.5; p = 0.006) as compared to NFY group. Retinol binding protein (mean diff: 0.05; 95% CI: 0.002-0.09; p = 0.04) and iodine levels (mean difference: 39.87; 95% CI: 20.39-59.35; p < 0.001) decreased between base and end-line but the decrease was significantly less in the FY group. Compared to NFY, the FY group had better height gain velocity (mean diff: 0.32; 95% CI: 0.05-0.60; p = 0.02) and height-for-age z-scores (mean diff: 0.18; 95% CI: 0.02-0.33; p = 0.03). There was no difference in weight gain velocity, weight-for-age z-scores or Body Mass Index z-scores. In the absence of iron deficiency at baseline the impact on iron status would not be expected to be observed and hence cannot be evaluated. Improved Hb concentrations in the absence of a change in iron status suggest improved utilization of iron possibly due to vitamin A and zinc availability. Fortification improved height gain without affecting weight gain. ClinicalTrial.gov: NCT00980733.


Proton-sensitive transitions of renal type II Na(+)-coupled phosphate cotransporter kinetics.
In the kidney proximal tubule, acidification of the glomerular filtrate leads to an inhibition of inorganic phosphate (P(i)) reabsorption by type II Na(+)-coupled cotransporters (NaPi-II). As external pH also alters the divalent/monovalent P(i) ratio, it has been difficult to separate putative proton interactions with the cotransporter from direct titration of divalent P(i), the preferred species transported. To distinguish between these possibilities and identify pH-sensitive transitions in the cotransport cycle, the pH-dependent kinetics of two NaPi-II isoforms, expressed in Xenopus laevis oocytes, were investigated electrophysiologically. At -50 mV, both isoforms showed >70% suppression of electrogenic response for an external pH change from 8.0 to 6.2, not attributable to titration of divalent P(i). This was accompanied by a progressive removal of steady-state voltage dependence. The NaPi-II-related uncoupled slippage current was unaffected by a pH change from 7.4 to 6.2, with no shift in the reversal potential, which suggested that protons do not function as substrate. The voltage-dependence of pre-steady-state relaxations was shifted to depolarizing potentials in 100 mM and 0 mM Na(ext)(+) and two kinetic components were resolved, the slower of which was pH-dependent. The changes in kinetics are predicted by a model in which protons interact with the empty carrier and final Na(+) binding step.


Amphetamine modulates human incentive processing.
Research suggests that psychostimulants can physiologically alter dopamine kinetics in the ventral striatum (VS) and psychologically enhance mood and attention. Using event-related functional magnetic resonance imaging (fMRI), we conducted a within-subject, double-blind, placebo (PLAC)-controlled study of the effects of oral dextroamphetamine (AMPH, 0.25 mg/kg) treatment on brain activity and affect during incentive processing. In two counterbalanced scanning sessions 60-180 min after ingesting AMPH or PLAC, 8 healthy volunteers played a game involving anticipation and receipt of monetary gains and losses. Group and volume of interest analyses suggested that by enhancing tonic over phasic activation, AMPH treatment "equalized" levels of VS activity and positive arousal during anticipation of both gain and loss. These findings suggest that therapeutic effects of amphetamine on incentive processing may involve reducing the difference between anticipation of gains and losses.


Predicting emergency department admissions.
To develop and validate models to predict emergency department (ED) presentations and hospital admissions for time and day of the year. Initial model development and validation was based on 5 years of historical data from two dissimilar hospitals, followed by subsequent validation on 27 hospitals representing 95% of the ED presentations across the state. Forecast accuracy was assessed using the mean average percentage error (MAPE) between forecasts and observed data. The study also determined a daily sample size threshold for forecasting subgroups within the data. Presentations to the ED and subsequent admissions to hospital beds are not random and can be predicted. Forecast accuracy worsened as the forecast time intervals became smaller: when forecasting monthly admissions, the best MAPE was approximately 2%, for daily admissions, 11%; for 4-hourly admissions, 38%; and for hourly admissions, 50%. Presentations were more easily forecast than admissions (daily MAPE ∼7%). When validating accuracy at additional hospitals, forecasts for urban facilities were generally more accurate than regional forecasts (accuracy is related to sample size). Subgroups within the data with more than 10 admissions or presentations per day had forecast errors statistically similar to the entire dataset. The study also included a software implementation of the models, resulting in a data dashboard for bed managers. Valid ED prediction tools can be generated from access to de-identified historic data, which may be used to assist elective surgery scheduling and bed management. The paper provides forecasting performance levels to guide similar studies.


Identification of gene-based responses in human blood cells exposed to alpha particle radiation.
The threat of a terrorist-precipitated nuclear event places humans at danger for radiological exposures. Isotopes which emit alpha (α)-particle radiation pose the highest risk. Currently, gene expression signatures are being developed for radiation biodosimetry and triage with respect to ionizing photon radiation. This study was designed to determine if similar gene expression profiles are obtained after exposures involving α-particles. Peripheral blood mononuclear cells (PBMCs) were used to identify sensitive and robust gene-based biomarkers of α-particle radiation exposure. Cells were isolated from healthy individuals and were irradiated at doses ranging from 0-1.5 Gy. Microarray technology was employed to identify transcripts that were differentially expressed relative to unirradiated cells 24 hours post-exposure. Statistical analysis identified modulated genes at each of the individual doses. Twenty-nine genes were common to all doses with expression levels ranging from 2-10 fold relative to control treatment group. This subset of genes was further assessed in independent complete white blood cell (WBC) populations exposed to either α-particles or X-rays using quantitative real-time PCR. This 29 gene panel was responsive in the α-particle exposed WBCs and was shown to exhibit differential fold-changes compared to X-irradiated cells, though no α-particle specific transcripts were identified. Current gene panels for photon radiation may also be applicable for use in α-particle radiation biodosimetry.


Standardization of medium for culturing Lyme disease spirochetes.
To standardize the procedure for isolating and culturing Lyme disease spirochetes, we modified the composition of the medium generally used for this purpose (BSK-II) and developed a system for its distribution. This medium contains no gelatin or agarose, and various components are used in proportions that differ from those in BSK-II. Each of the major proteinacious components was screened by substitution in samples of the complete product. The final medium was evaluated for the capacity to grow related spirochetes including Borrelia burgdorferi N40, Guilford, and JD-1 as well as strains of Borrelia hermsii (HS-1) and of Borrelia coriaceae (CO53). Each isolate developed from inocula containing as few as one to five organisms. Doubling time of B. burgdorferi during log-phase growth at 37 degrees C was 10 to 12 h. Lyme disease spirochetes were isolated in this medium from ear punch biopsies and dermal aspirates from naturally infected mice and rabbits, from dermal biopsies from a human patient, and by sampling field-collected deer ticks (Ixodes dammini). Cultured spirochetes remained infective to mice and to ticks. The medium can be stored at -20 degrees C or lower temperatures for at least 8 months without effect on its ability to support growth of small inocula to densities exceeding 10(8) spirochetes per ml. Lyme disease spirochetes remained infective to mice after being stored at -80 degrees C in this medium for at least 8 months. We anticipate that the availability of this standardized medium (Sigma Chemical Co.), supplemented with prescreened rabbit serum, will facilitate comparison of research results between laboratories and may eventually permit definitive clinical diagnosis of Lyme disease based on demonstration of the pathogen. The standardized medium is designated BSK-H.


Detection of mRNA by reverse transcription-PCR as an indicator of viability in Escherichia coli cells.
The relationship between the detection of mRNA and cellular viability in Escherichia coli was investigated in cells killed by heat or ethanol. Reverse transcription-PCR (RT-PCR) methods were developed for detecting mRNA from rpoH, groEL, and tufA genes. mRNA from all three genes was detected immediately after the cells had been killed by heat or ethanol but gradually disappeared with time when dead cells were held at room temperature. In heat-killed cells, some mRNA targets became undetectable after 2 to 16 h, whereas after ethanol treatment, mRNA was still detected after 16 h. In contrast, 16S rRNA was detected by RT-PCR in all samples containing dead cells and did not disappear during a subsequent incubation of 16 h at room temperature. Of the different types of nucleic acid, mRNA is the most promising candidate for an indicator of viability in bacteria, but its persistence in dead cells depends on the inactivating treatment and subsequent holding conditions.


Characterization of Escherichia coli O157:H7 strains from contaminated raw beef trim during "high event periods".
The development and implementation of effective antimicrobial interventions by the beef processing industry in the United States have dramatically reduced the incidence of beef trim contamination by Escherichia coli O157:H7. However, individual processing plants still experience sporadic peaks in contamination rates where multiple E. coli O157:H7-positive lots are clustered in a short time frame. These peaks have been referred to as "high event periods" (HEP) of contamination. The results reported here detail the characterization of E. coli O157:H7 isolates from 21 HEP across multiple companies and processing plants to gain insight regarding the mechanisms causing these incidents. Strain genotypes were determined by pulsed-field gel electrophoresis, and isolates were investigated for characteristics linking them to human illness. Through these analyses, it was determined that individual HEP show little to no diversity in strain genotypes. Hence, each HEP has one strain type that makes up most, if not all, of the contamination. This is shown to differ from the genotypic diversity of E. coli O157:H7 found on the hides of cattle entering processing plants. In addition, it was found that a large proportion (81%) of HEP are caused by strain types associated with human illness. These results pose a potential challenge to the current model for finished product contamination during beef processing.


The malaria parasite monitored by photoacoustic spectroscopy.
Noninvasive photoacoustic spectroscopy was used to study the malaria parasites Plasmodium chabaudi and Plasmodium berghei, their pigment, and ferriprotoporphyrin IX, which is a by-product of the hemoglobin that the parasite ingests. The results indicate that the pigment consists of ferriprotophorphyrin self-aggregates and a noncovalent complex of ferriprotoporphyrin and protein. Spectra of chloroquine-treated parasites reveal in situ interaction between the drug and ferriprotoporphyrin. Chloroquine-resistant parasites, readily distinguishable by this method, appear to degrade hemoglobin only partially.


Changes in Awake Cystometry and Expression of Bladder β-adrenoceptors after Partial Bladder Outlet Obstruction in Male Rats.
To explore possible changes in awake cystometry and expression of beta-adrenoceptors (ARs) as a cause for bladder dysfunction in a male rat model of partial bladder outlet obstruction (pBOO). Awake cystometry was performed in rats with pBOO (16) and sham-operated rats (16), 8 weeks after the operation. The expression of mRNA and protein of β-ARs was assessed by real-time PCR and western blot. The bladders with pBOO (1030mg) were increased compared to those in control rats (230mg). In the cystometric studies, the maximum intravesical pressure significantly increased in the pBOO group compared to control group rats (p=0.001). The time to reach maximal intravesical pressure during micturition in the pBOO group was significantly longer than the sham group (p=0.003). The frequency of non-void contraction in the pBOO group was significantly more than the sham group (p=0.006). The mRNA expressions of β2- and 3-ARs were increased insignificantly in pBOO group compared to sham group. The data of pBOO group expressed as folds of corresponding expression in sham group were 1.28 and 1.46 respectively in β2- and 3-ARs. Compared to the sham groups, the density of 60Kda protein band recognized by β2-AR antibodies and the density of 45Kda protein band recognized by β3-AR antibodies were higher in the bladder from pBOO group rats. PBOO of male rats increase the maximal intravesical pressure and contraction time during micturition and the frequency of non-void contraction as well as weight of bladder. The expression of β2- and 3-ARs subtypes was increased insignificantly compared to sham operated group. This study demonstrates that the changes of cystometric or non-void contraction parameters in pBOO is one of the pathophysiologic processes potentially associated with the alterations of bladder β-ARs.


Internal thoracic artery grafting in the elderly patient undergoing coronary artery bypass grafting: room for process improvement?
The acute and long-term benefits of internal thoracic artery grafting are clear in younger patients undergoing coronary artery bypass grafting. The elderly, however, face higher surgical risks and have shorter life expectancy, and thus the use of internal thoracic artery grafting in this age group has been debated. This study examined the use, complication risks, and operative (30-day) mortality associated with internal thoracic artery grafting in patients 75 years of age and older. Between 1996 and 1999, 522,656 patients in the Society of Thoracic Surgeons National Cardiac Database underwent primary, nonemergency-salvage coronary artery bypass grafting; of these, 99,942 were 75 years of age or older. The influence of internal thoracic artery use on operative mortality and 5 major complications in this elderly group was examined by means of (1) risk adjustment (adjusting for 28 baseline risk factors and site) and (2) a treatment propensity score analysis that compares patients with similar baseline likelihood for receiving an internal thoracic artery graft. In the National Cardiac Database 77.4% of patients aged 75 to 84 years received an internal thoracic artery graft compared with 93.5% for those aged 55 years or less. In this elderly group use of the internal thoracic artery was strongly associated with decreased operative mortality (unadjusted mortality, 6.20% vs. 4.05%; P <.0001) that persisted after controlling for baseline risk and provider effects (adjusted odds ratio, 0.85; 95% confidence intervals, 0.79-0.91). This mortality benefit was seen among those with low-to-high baseline propensity for receiving an internal thoracic artery graft. Use of the internal thoracic artery in elderly patients undergoing coronary artery bypass grafting provides an acute survival benefit. This benefit is similar to that seen in younger patients and persists after adjusting for both patient and provider selection factors. The internal thoracic artery appears to be underused in elderly patients undergoing bypass grafting and is a potential area for quality improvement.


Brief intervention for problem drug use in safety-net primary care settings: a randomized clinical trial.
Although brief intervention is effective for reducing problem alcohol use, few data exist on its effectiveness for reducing problem drug use, a common issue in disadvantaged populations seeking care in safety-net medical settings (hospitals and community health clinics serving low-income patients with limited or no insurance). To determine whether brief intervention improves drug use outcomes compared with enhanced care as usual. A randomized clinical trial with blinded assessments at baseline and at 3, 6, 9, and 12 months conducted in 7 safety-net primary care clinics in Washington State. Of 1621 eligible patients reporting any problem drug use in the past 90 days, 868 consented and were randomized between April 2009 and September 2012. Follow-up participation was more than 87% at all points. Participants received a single brief intervention using motivational interviewing, a handout and list of substance abuse resources, and an attempted 10-minute telephone booster within 2 weeks (n = 435) or enhanced care as usual, which included a handout and list of substance abuse resources (n = 433). The primary outcomes were self-reported days of problem drug use in the past 30 days and Addiction Severity Index-Lite (ASI) Drug Use composite score. Secondary outcomes were admission to substance abuse treatment; ASI composite scores for medical, psychiatric, social, and legal domains; emergency department and inpatient hospital admissions, arrests, mortality, and human immunodeficiency virus risk behavior. Mean days used of the most common problem drug at baseline were 14.40 (SD, 11.29) (brief intervention) and 13.25 (SD, 10.69) (enhanced care as usual); at 3 months postintervention, means were 11.87 (SD, 12.13) (brief intervention) and 9.84 (SD, 10.64) (enhanced care as usual) and not significantly different (difference in differences, β = 0.89 [95% CI, -0.49 to 2.26]). Mean ASI Drug Use composite score at baseline was 0.11 (SD, 0.10) (brief intervention) and 0.11 (SD, 0.10) (enhanced care as usual) and at 3 months was 0.10 (SD, 0.09) (brief intervention) and 0.09 (SD, 0.09) (enhanced care as usual) and not significantly different (difference in differences, β = 0.008 [95% CI, -0.006 to 0.021]). During the 12 months following intervention, no significant treatment differences were found for either variable. No significant differences were found for secondary outcomes. A one-time brief intervention with attempted telephone booster had no effect on drug use in patients seen in safety-net primary care settings. This finding suggests a need for caution in promoting widespread adoption of this intervention for drug use in primary care. clinicaltrials.gov Identifier: NCT00877331.


The mechanism of rare earth incorporation in solution doping process.
The mechanism involved during solution doping process has been systematically investigated by correlating the soot characteristics and solution parameters with the amount of rare earth (RE) incorporated in the core of optical fiber. Experiments show that the amount of RE incorporation may be controlled with better precision by adjusting Al ion concentration in the soaking solution. A model has been developed on the basis of cooperative adsorption mechanism correlating different parameters in the overall process. Theoretical estimation shows good agreement with the experimental results and can be used to predict the extent of RE incorporation for any composition if the soot layer characteristics are known.


[DNA polymerase gamma in the cells of loach embryos localized in mitochondria].
The homogenate of loach (Misgurnus fossilis) embryonic cells was fractionated by differential centrifugation into fractions, in which the activities of DNA polymerases alpha and gamma were measured. About half of the total alpha-polymerase activity was retained in the postmicrosomal supernatant. The activity of gamma-polymerase in the supernatant did not exceed 2% of the total activity. The specific activity of gamma-polymerase was maximal in the mitochondrial pellet. The enzyme activity in the nuclear pellet was proportional to the activity of mitochondrial enzymes, cytochrome oxidase and KCN-insensitive superoxide dismutase. The nuclei were separated from gamma-polymerase-containing fraction by ultracentrifugation in a 10-50% sucrose density gradient. The material of this fraction was identical to the mitochondria in terms of buoyant density, morphology and enzymatic properties. Thus, in loach embryos the fraction of gamma-polymerase associated with the nuclei is absent; the total gamma-polymerase activity is localized in the mitochondria.


Functional characterization of TRPV4 as an osmotically sensitive ion channel in porcine articular chondrocytes.
Transient receptor potential vanilloid 4 (TRPV4) is a Ca(2+)-permeable channel that can be gated by tonicity (osmolarity) and mechanical stimuli. Chondrocytes, the cells in cartilage, respond to their osmotic and mechanical environments; however, the molecular basis of this signal transduction is not fully understood. This study was undertaken to demonstrate the presence and functionality of TRPV4 in chondrocytes. TRPV4 protein expression was measured by immunolabeling and Western blotting. In response to TRPV4 agonist/antagonists, osmotic stress, and interleukin-1 (IL-1), changes in Ca(2+) signaling, cell volume, and prostaglandin E(2) (PGE(2)) production were measured in porcine chondrocytes using fluorescence microscopy, light microscopy, or immunoassay, respectively. TRPV4 was expressed abundantly at the RNA and protein levels. Exposure to 4alpha-phorbol 12,13-didecanoate (4alphaPDD), a TRPV4 activator, caused Ca(2+) signaling in chondrocytes, which was blocked by the selective TRPV4 antagonist, GSK205. Blocking TRPV4 diminished the chondrocytes' response to hypo-osmotic stress, reducing the fraction of Ca(2+) responsive cells, the regulatory volume decrease, and PGE(2) production. Ca(2+) signaling was inhibited by removal of extracellular Ca(2+) or depletion of intracellular stores. Specific activation of TRPV4 restored the defective regulatory volume decrease caused by IL-1. Chemical disruption of the primary cilium eliminated Ca(2+) signaling in response to either 4alphaPDD or hypo-osmotic stress. Our findings indicate that TRPV4 is present in articular chondrocytes, and chondrocyte response to hypo-osmotic stress is mediated by this channel, which involves both an extracellular Ca(2+) and intracellular Ca(2+) release. TRPV4 may also be involved in modulating the production or influence of proinflammatory molecules in response to osmotic stress.


Fatal human herpesvirus 6-associated encephalitis in two boys with underlying POLG mitochondrial disorders.
Human herpesvirus 6 is a significant cause of the febrile illness roseola infantum in young children. Infection with human herpesvirus 6 typically causes a self-limited febrile illness but occasionally is associated with central nervous system manifestations, including febrile seizures and encephalitis. Host factors associated with severe manifestations of human herpesvirus 6-associated neurological disease remain poorly characterized. We report two previously healthy young boys with human herpesvirus 6-associated encephalitis who developed a progressive, and ultimately fatal, encephalopathy with refractory movement disorder concurrent with acquisition of acute human herpesvirus 6 infection. Both children were treated with the antiviral ganciclovir without improvement of their neurological symptoms, although quantitative human herpesvirus 6 polymerase chain reaction of cerebrospinal fluid and/or blood confirmed a decline in viral load with treatment. The clinical course in both cases was most consistent with Alpers-Huttenlocher syndrome, given the intractable seizures, developmental regression, and, ultimately, death due to liver and renal failure. In support of this, postmortem analysis identified both children to be compound heterozygous for mutations in the mitochondrial polymerase γ gene, POLG. POLG mutations are associated with Alpers-Huttenlocher syndrome; however, no prior studies have examined the role of acute human herpesvirus 6 infection in these patients presenting with severe neurological disease. It is possible the POLG mutation phenotype was unmasked and/or exacerbated by human herpesvirus 6 infection in these two patients, potentially contributing to a more rapid clinical deterioration. This report provides new insight into a previously unrecognized association between POLG mutations and poor neurological outcome after human herpesvirus 6 infection.


The mechanism of adenosine production in rat polymorphonuclear leucocytes.
Adenosine production in intact rat polymorphonuclear leucocytes was studied during 2-deoxyglucose-induced ATP catabolism. A cell-free system containing the cytosolic 5'-nucleotidase (EC 188.8.131.52) as the only phosphohydrolase was also studied. The rate of adenosine formation in both intact cells and the cell-free system showed a similar dependence on energy charge (([ATP] + 1/2 [ADP]/([ATP] + [ADP] + [AMP])), being maximal only at values close to 0.8. Sufficient cytosolic 5'-nucleotidase was present in intact cells to explain the observed rate of adenosine formation. We conclude that the cytosolic 5'-nucleotidase is responsible for adenosine production in rat polymorphonuclear leucocytes. This mechanism provides a direct biochemical link between the energy status of a cell and the rate of adenosine formation.


Evaluation of the burden of unsuspected pulmonary tuberculosis and co-morbidity with non-communicable diseases in sputum producing adult inpatients.
A high burden of tuberculosis (TB) occurs in sub-Saharan African countries and many cases of active TB and drug-resistant TB remain undiagnosed. Tertiary care hospitals provide an opportunity to study TB co-morbidity with non-communicable and other communicable diseases (NCDs/CDs). We evaluated the burden of undiagnosed pulmonary TB and multi-drug resistant TB in adult inpatients, regardless of their primary admission diagnosis, in a tertiary referral centre. In this prospective study, newly admitted adult inpatients able to produce sputum at the University Teaching Hospital, Lusaka, Zambia, were screened for pulmonary TB using fluorescent smear microscopy and automated liquid culture. The burden of pulmonary TB, unsuspected TB, TB co-morbidity with NCDs and CDs was determined. Sputum was analysed from 900 inpatients (70.6% HIV infected) 277 (30.8%) non-TB suspects, 286 (31.8%) TB suspects and 337 (37.4%) were already receiving TB treatment. 202/900 (22.4%) of patients had culture confirmed TB. TB co-morbidity was detected in 20/275 (7.3%) NCD patients, significantly associated with diabetes (P = 0.006, OR 6.571, 95%CI: 1.706-25.3). 27/202 (13.4%) TB cases were unsuspected. There were 18 confirmed cases of MDR-TB, 5 of which were unsuspected. A large burden of unsuspected pulmonary TB co-morbidity exists in inpatients with NCDs and other CDs. Pro-active sputum screening of all inpatients in tertiary referral centres in high TB endemic countries is recommended. The scale of the problem of undiagnosed MDR-TB in inpatients requires further study.


Knowledge of patients on safe medication use in relation to nonsteroidal anti-inflammatory drugs.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely prescribed and used medications that are used as analgesic, antipyretic, or anti-inflammatory agents, but have a high risk of adverse effects. The main purpose of this study is to evaluate the knowledge of patients toward the risks and safe usage of NSAIDs. A descriptive cross-sectional study was conducted among patients who attended orthopedic clinics at King Khalid University Hospital in Riyadh, Saudi Arabia, during December 2017 to February 2018. A self-administered questionnaire comprising three sections [sociodemographic information, patients' knowledge of NSAIDs, and the attitude toward receiving information about NSAIDs' adverse drug reactions (ADRs)] was used. The attitude was measured with a 5-point Likert scale. The mean duration of NSAIDs usage was 7.3 ± 6.9 years. Only 45% of the respondents used NSAIDs daily, while 38% said they only used NSAIDs as needed. Nearly one-quarter of patients (25.5%) were counseled by a healthcare provider on the risks of NSAIDs. Almost 94% and 90.5% of the study subjects agreed that physicians and pharmacists, respectively, should play an essential role in providing information regarding ADRs to their patients. More than half of the respondents claimed that information about ADRs might lead them to discontinue their medicines. More than one-third of respondents stated that their healthcare setting provided them with insufficient knowledge of ADRs. Counseling of patients may diminish potentially inappropriate use and increase risk awareness. Healthcare providers can improve their role in identifying and counseling patients on NSAIDs.