Association of Japanese encephalitis virus NS3 protein with microtubules and tumour susceptibility gene 101 (TSG101) protein.
Previously reported findings by our group showed that non-structural protein 3 (NS3) of Japanese encephalitis virus (JEV) was localized mainly in the JEV-induced convoluted membrane (CM), which has been proposed to originate from rough endoplasmic reticulum (rER), Golgi apparatus or the trans-Golgi network (TGN), and serves as a reservoir for viral proteins during virus assembly. Earlier findings indicated that NS3 of Kunjin virus interacts with microtubules. In addition, one of the Golgi-associated proteins, tumour susceptibility protein 101 (TSG101), associates with microtubules and is required for budding of retroviral particles. To clarify the association of NS3 with microtubules or with TSG101 during JEV assembly, we applied immunofluorescence, co-immunoprecipitation and immunoelectron microscopic methods. Virus infection, as well as transfection with an NS2B-NS3 expression plasmid, induced microtubule rearrangement. When cells were treated with colchicine, which interferes with microtubule polymerization, NS3 still associated with tubulin and TSG101. Furthermore, tubulin and TSG101 were co-localized with NS3 in the CM by immunogold labelling. Our observations indicate that microtubules and TSG101 associate with NS3, which is incorporated into the JEV-induced structure during JEV replication.


Khyâl attacks: a key idiom of distress among traumatized cambodia refugees.
Traumatized Cambodian refugees with PTSD often complain of khyâl attacks. The current study investigates khyâl attacks from multiple perspectives and examines the validity of a model of how khyâl attacks are generated. The study found that khyâl attacks had commonly been experienced in the previous 4 weeks and that their severity was strongly correlated with the severity of PTSD (PTSD Checklist). It was found that khyâl attacks were triggered by various processes--such as worry, trauma recall, standing up, going to a mall--and that khyâl attacks almost always met panic attack criteria. It was also found that during a khyâl attack there was great fear that death might occur from bodily dysfunction. It was likewise found that a complex nosology of khyâl attacks exists that rates the attacks on a scale of severity, that the severity determines how the khyâl attacks should be treated and that those treatments are often complex. As illustrated by the article, khyâl attacks constitute a key aspect of trauma ontology in this group, a culturally specific experiencing of anxiety and trauma-related disorder. The article also contributes to the study of trauma somatics, that is, to the study of how trauma results in specific symptoms in a specific cultural context, showing that a key part of the trauma-somatic reticulum is often a cultural syndrome.


Preschoolers will drink their GREENS! Children accept, like, and drink novel smoothies containing dark green vegetables (DGVs).
Dark green vegetables (DGVs; e.g., spinach) are a nutrient rich source of essential vitamins and minerals; yet, children's intakes of DGVs fall well below dietary recommendations and creative solutions are needed. This study describes preschoolers (3-5 y) willingness to taste, liking, and intake of fruit-based smoothies containing DGVs (i.e., spinach, collards, kale), commonly referred to as "green smoothies," and explores individual differences in children's eating responses. Using a between-subjects design, preschoolers were randomized to either a FRUIT ONLY smoothie condition (n = 36) or FRUIT+DGV smoothie condition (n = 32). Children's acceptance and intake were collected in one tasting session and one ad libitum snack session, respectively. Parents reported on child food pickiness, food responsiveness, and approach, and children's intake of fruits and DGVs. Children self-reported on previous experience with the study fruits and DGVs. The initial tasting session revealed that the majority of children (84.3%) in the FRUIT+DGV condition willingly tasted all five green smoothies and rated the green smoothies as moderately liked (2.3 ± 0.1). Children in the FRUIT+DGV condition consumed 225.7 ± 31.4 g (9.0 ± 1.3 oz; 1.1 ± 0.2 cups; 91.9 ± 12.9 kcals) of their most preferred green smoothie, providing 18.3 ± 3.7 g (or 0.7 ± 0.1 cups) of DGVs. Children's willingness to try, liking, and intake did not differ by smoothie condition. Individual differences in children's intake are reported. In conclusion, children were willing to try fruit smoothies supplemented with DGVs. Children rated the green smoothies as moderately-liked and children's intake during snack met 31% of their weekly USDA recommendations for DGVs. Adding DGVs to fruit-based smoothies may compliment other effective feeding strategies for increasing children's vegetable consumption.


Sodium ((23)Na) ultra-short echo time imaging in the human brain using a 3D-Cones trajectory.
Sodium magnetic resonance imaging ((23)Na-MRI) of the brain has shown changes in (23)Na signal as a hallmark of various neurological diseases such as stroke, Alzheimer's disease, Multiple Sclerosis and Huntington's disease. To improve scan times and image quality, we have implemented the 3D-Cones (CN) sequence for in vivo (23)Na brain MRI. Using signal-to-noise (SNR) as a measurement of sequence performance, CN is compared against more established 3D-radial k-space sampling schemes featuring cylindrical stack-of-stars (SOS) and 3D-spokes kooshball (KB) trajectories, on five healthy volunteers in a clinical setting. Resolution was evaluated by simulating the point-spread-functions (PSFs) and experimental measures on a phantom. All sequences were shown to have a similar SNR arbitrary units (AU) of 6-6.5 in brain white matter, 7-9 in gray matter and 17-18 AU in cerebrospinal fluid. SNR between white and gray matter were significantly different for KB and CN (p = 0.046 and <0.001 respectively), but not for SOS (p = 0.1). Group mean standard deviations were significantly smaller for CN (p = 0.016). Theoretical full-width at half-maximum linewidth of the PSF for CN is broadened by only 0.1, compared to 0.3 and 0.8 pixels for SOS and KB respectively. Actual image resolution is estimated as 8, 9 and 6.3 mm for SOS, KB and CN respectively. The CN sequence provides stronger tissue contrast than both SOS and KB, with more reproducible SNR measurements compared to KB. For CN, a higher true resolution in the same amount of time with no significant trade-off in SNR is achieved. CN is therefore more suitable for (23)Na-MRI in the brain.


MM Algorithms For Variance Components Models.
Variance components estimation and mixed model analysis are central themes in statistics with applications in numerous scientific disciplines. Despite the best efforts of generations of statisticians and numerical analysts, maximum likelihood estimation and restricted maximum likelihood estimation of variance component models remain numerically challenging. Building on the minorization-maximization (MM) principle, this paper presents a novel iterative algorithm for variance components estimation. Our MM algorithm is trivial to implement and competitive on large data problems. The algorithm readily extends to more complicated problems such as linear mixed models, multivariate response models possibly with missing data, maximum a posteriori estimation, and penalized estimation. We establish the global convergence of the MM algorithm to a Karush-Kuhn-Tucker (KKT) point and demonstrate, both numerically and theoretically, that it converges faster than the classical EM algorithm when the number of variance components is greater than two and all covariance matrices are positive definite.


Rastering strategy for screening and centring of microcrystal samples of human membrane proteins with a sub-10 microm size X-ray synchrotron beam.
Crystallization of human membrane proteins in lipidic cubic phase often results in very small but highly ordered crystals. Advent of the sub-10 microm minibeam at the APS GM/CA CAT has enabled the collection of high quality diffraction data from such microcrystals. Herein we describe the challenges and solutions related to growing, manipulating and collecting data from optically invisible microcrystals embedded in an opaque frozen in meso material. Of critical importance is the use of the intense and small synchrotron beam to raster through and locate the crystal sample in an efficient and reliable manner. The resulting diffraction patterns have a significant reduction in background, with strong intensity and improvement in diffraction resolution compared with larger beam sizes. Three high-resolution structures of human G protein-coupled receptors serve as evidence of the utility of these techniques that will likely be useful for future structural determination efforts. We anticipate that further innovations of the technologies applied to microcrystallography will enable the solving of structures of ever more challenging targets.


Drugs with anticholinergic effects and cognitive impairment, falls and all-cause mortality in older adults: A systematic review and meta-analysis.
The aim was to investigate associations between drugs with anticholinergic effects (DACEs) and cognitive impairment, falls and all-cause mortality in older adults. A literature search using CINAHL, Cochrane Library, Embase and PubMed databases was conducted for randomized controlled trials, prospective and retrospective cohort and case-control studies examining the use of DACEs in subjects ≥65 years with outcomes on falls, cognitive impairment and all-cause mortality. Retrieved articles were published on or before June 2013. Anticholinergic exposure was investigated using drug class, DACE scoring systems (anticholinergic cognitive burden scale, ACB; anticholinergic drug scale, ADS; anticholinergic risk scale, ARS; anticholinergic component of the drug burden index, DBIAC ) or assessment of individual DACEs. Meta-analyses were performed to pool the results from individual studies. Eighteen studies fulfilled the inclusion criteria (total 124 286 participants). Exposure to DACEs as a class was associated with increased odds of cognitive impairment (OR 1.45, 95% CI 1.16, 1.73). Olanzapine and trazodone were associated with increased odds and risk of falls (OR 2.16, 95% CI 1.05, 4.44; RR 1.79, 95% CI 1.60, 1.97, respectively), but amitriptyline, paroxetine and risperidone were not (RR 1.73, 95% CI 0.81, 2.65; RR 1.80, 95% CI 0.81, 2.79; RR 1.39, 95% CI 0.59, 3.26, respectively). A unit increase in the ACB scale was associated with a doubling in odds of all-cause mortality (OR 2.06, 95% CI 1.82, 2.33) but there were no associations with the DBIAC (OR 0.88, 95% CI 0.55, 1.42) or the ARS (OR 3.56, 95% CI 0.29, 43.27). Certain individual DACEs or increased overall DACE exposure may increase the risks of cognitive impairment, falls and all-cause mortality in older adults.


Time-Resolved Temperature-Jump Infrared Spectroscopy at a High Repetition Rate.
Time-resolved temperature-jump infrared absorption spectroscopy at a 0.5 to 1 kHz repetition rate is presented. A 1 kHz neodymium-doped yttrium aluminum garnet (Nd:YAG) laser pumping an optical parametric oscillator provided >70 µJ, 3.75 µm pump pulses, which delivered a temperature jump via excitation of the O-D stretch of a D2O solution. A 10 kHz train of mid-infrared probe pulses was used to monitor spectral changes following the temperature jump. Calibration with trifluoroacetic acid solution showed that a temperature jump of 10 K lasting for tens of microseconds was achieved, sufficient to observe fast processes in functionally relevant biomolecular mechanisms. Modeling of heating profiles across ≤10 µm path length cells and subsequent cooling dynamics are used to describe the initial <100 ns cooling at the window surface and subsequent, >10 µs cooling dynamics of the bulk solution.


Comprehensive men's health and male infertility.
There is increasing evidence that male infertility may be a harbinger of comorbid medical illness. Existing studies have shed light on associations between infertility and the prevalence of cardiovascular, metabolic, and oncologic disease, along with rates of hospitalization and overall mortality. Although theorized mechanisms include genetic, developmental, and behavioral precipitants, the exact nature of these associations remains unclear and warrants further investigation.


Direct sequencing of SARS-coronavirus S and N genes from clinical specimens shows limited variation.
Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged, in November 2002, as a novel agent causing severe respiratory illness. To study sequence variation in the SARS-CoV genome, we determined the nucleic acid sequence of the S and N genes directly from clinical specimens from 10 patients--1 specimen with no matched SARS-CoV isolate, from 2 patients; multiple specimens from 3 patients; and matched clinical-specimen/cell-culture-isolate pairs from 6 patients. We identified 3 nucleotide substitutions that were most likely due to natural variation and 2 substitutions that arose after cell-culture passage of the virus. These data demonstrate the overall stability of the S and N genes of SARS-CoV over 3 months during which a minimum of 4 generations for transmission events occurred. These findings are a part of the expanding investigation of the evolution of how this virus adapts to a new host.


Respiratory muscle activity related to flow and lung volume in preterm infants compared with term infants.
Infants with chronic lung disease (CLD) have a capacity to maintain functional lung volume despite alterations to their lung mechanics. We hypothesize that they achieve this by altering breathing patterns and dynamic elevation of lung volume, leading to differences in the relationship between respiratory muscle activity, flow and lung volume. Lung function and transcutaneous electromyography of the respiratory muscles (rEMG) were measured in 20 infants with CLD and in 39 healthy age-matched controls during quiet sleep. We compared coefficient of variations (CVs) of rEMG and the temporal relationship of rEMG variables, to flow and lung volume [functional residual capacity (FRC)] between these groups. The time between the start of inspiratory muscle activity and the resulting flow (tria)--in relation to respiratory cycle time--was significantly longer in infants with CLD. Although FRC had similar associations with tria and postinspiratory activity (corrected for respiratory cycle time), the CV of the diaphragmatic rEMG was lower in CLD infants (22.6 versus 31.0%, p = 0.030). The temporal relationship of rEMG to flow and FRC and the loss of adaptive variability provide additional information on coping mechanisms in infants with CLD. This technique could be used for noninvasive bedside monitoring of CLD.


Applications of Raman spectroscopy in the development of cell therapies: state of the art and future perspectives.
Therapies based on injecting living cells into patients offer a huge potential to cure many degenerative and deadly diseases, with hundreds of clinical trials ongoing. Due to their complex nature, a basic understanding of their biochemical and functional characteristics, how to manufacture them for safe and efficacious therapy, and how to effectively implement them in clinical settings are very challenging. Raman spectroscopy could provide an information-rich, non-invasive, non-destructive analytical method to complement the use of conventional sample-based, infrequent and destructive biochemical assays typically employed to analyze and validate the quality of therapeutic cells. This article provides an overview of the current state of emerging cell therapies, and then reviews the related Raman spectroscopic state of the art analysis of human cells. This includes spectroscopic data processing considerations, the scope offered by technical variants of Raman spectroscopy, and analytical difficulties encountered by spectroscopists working with therapeutic cells. Finally, we outline a number of salient challenges as cell therapy products are translated from the laboratory to the clinic, and propose how Raman spectroscopy-based solutions could address these challenges.


Mapping episodic fluctuations in plasma LH in orchidectomized rats.
Reports of the frequency of pulsatile LH release in orchidectomized rats are surprisingly variable. Estimates of the period between LH pulses vary from 18.5 to 250 min in nine reports from six laboratories, published in 1981-1983. In these studies, blood samples were drawn at intervals ranging from 2.5 to 60 min. We examined the relationship between estimated LH pulse frequency and blood-sampling interval. Six castrated male rats were cannulated, and blood samples were drawn at 2.5-min intervals through 4 h. Plasma LH levels were determined by radioimmunoassay. Using the data obtained at 2.5-min intervals, we simulated blood-sampling intervals of 5, 7.5, 10, 12.5, and 15 min by sequentially deleting data points. The original and simulated data sets were analyzed by both the PULSAR and Cycle Detector computer programs. The results show that as the sampling interval increased from 2.5 to 15 min, the apparent period of pulsatile LH release rose steadily from about 20 min to about 100 min. We found a high, positive correlation between blood-sampling interval and the apparent period of LH pulses. Estimates of LH pulse frequency in castrated male rats vary directly with the frequency at which blood samples are taken. Sampling intervals greater than 5 min in orchidectomized rats yield an LH pulse period that is most likely exaggerated.


The Role of the Y Chromosome in Brain Function.
In mammals, sex differences are evident in many aspects of brain development, brain function and behaviour. Ultimately, such differences must arise from the differential sex chromosome complements in males and females: males inherit a single X chromosome and a Y chromosome, whilst females inherit two X chromosomes. One possible mechanism for sexual differentiation of the brain is via male-limited expression of genes on the small Y chromosome. Many Y-linked genes have been implicated in the development of the testes, and therefore could theoretically contribute to sexual differentiation of the brain indirectly, through influencing gonadal hormone production. Alternatively, Y-linked genes that are expressed in the brain could directly influence neural masculinisation. The present paper reviews evidence from human genetic studies and animal models for Y-linked effects (both direct and indirect) on neurodevelopment, brain function and behaviour. Besides enhancing our knowledge of the mechanisms underlying mammalian neural sexual differentiation, studies geared towards understanding the role of the Y chromosome in brain function will help to elucidate the molecular basis of sex-biased neuropsychiatric disorders, allowing for more selective sex-specific therapies.


Na+/H+ Exchanger Regulatory Factor 1 Mediates the Pathogenesis of Airway Inflammation in a Murine Model of House Dust Mite-Induced Asthma.
Na+/H+ exchanger regulatory factor 1 (NHERF1), a class I PDZ-binding protein, regulates G protein-coupled receptor signaling in some cell types. NHERF1 also functions as a scaffolding protein and activates non-G protein-coupled receptor signaling pathways, thereby contributing to the pathogenesis of various diseases. Although we have previously shown that NHERF1 regulates mast cell functions, there is little information regarding the role of NHERF1 in other immune cells. How NHERF1 regulates the pathogenesis of allergic disease such as asthma also remains unknown. In the current study, we show that NHERF1 promotes allergic airway inflammation in a house dust mite extract (HDME)-induced mouse model of asthma. Specifically, HDME-specific serum IgE levels, airway leukocyte numbers, and goblet cell hyperplasia were reduced in NHERF1+/- mice as compared with NHERF1+/+ mice. Interestingly, the gene expression of inflammatory (IL-17a, IL-25, and IL-33) as well as T helper 2 (Th2) cytokines (IL-4, IL-5, and IL-13) and several chemokines that recruit eosinophils, neutrophils, and lymphocytes were also decreased in the lungs of NHERF1+/- mice exposed to HDME. Consistent with these observations, microRNAs regulating mucus production, inflammation, Th2 effector functions, and IL-13 expression were increased in the lungs of HDME-treated NHERF1+/- mice. Overall, our studies reveal a unique role for NHERF1 in regulating asthma pathogenesis, and further elucidation of the mechanisms through which NHERF1 modulates allergic inflammation will lead to the development of new therapeutic strategies for asthma.


Identifying a role for Toll-like receptor 3 in the innate immune response to Chlamydia muridarum infection in murine oviduct epithelial cells.
Because epithelial cells are the major cell type productively infected with Chlamydia during genital tract infections, the overall goal of our research was to understand the contribution of infected epithelial cells to the host defense. We previously showed that Toll-like receptor 3 (TLR3) is the critical pattern recognition receptor in oviduct epithelial (OE) cells that is stimulated during Chlamydia infection, resulting in the synthesis of beta interferon (IFN-β). Here, we present data that implicates TLR3 in the expression of a multitude of other innate-inflammatory immune modulators including interleukin-6 (IL-6), CXCL10, CXCL16, and CCL5. We demonstrate that Chlamydia-induced expression of these cytokines is severely disrupted in TLR3-deficient OE cells, whereas Chlamydia replication in the TLR3-deficient cells is more efficient than in wild-type OE cells. Pretreatment of the TLR3-deficient OE cells with 50 U of IFN-β/ml prior to infection diminished Chlamydia replication and restored the ability of Chlamydia infection to induce IL-6, CXCL10, and CCL5 expression in TLR3-deficient OE cells; however, CXCL16 induction was not restored by IFN-β preincubation. Our findings were corroborated in pathway-focused PCR arrays, which demonstrated a multitude of different inflammatory genes that were defectively regulated during Chlamydia infection of the TLR3-deficient OE cells, and we found that some of these genes were induced only when IFN-β was added prior to infection. Our OE cell data implicate TLR3 as an essential inducer of IFN-β and other inflammatory mediators by epithelial cells during Chlamydia infection and highlight the contribution of TLR3 to the inflammatory cytokine response.


Intra-capsular complete tonsillectomy, a modification of surgical technique to eliminate delayed post-operative bleeding.
Pediatric tonsillectomy is a frequently performed procedure using a variety of surgical techniques. Despite decades of modern surgical experience and advances in tools and techniques, a consensus on which technique is superior for minimizing complications has not been established. In a 2014 study, the authors examined differing surgical techniques and devices to determine if there was a significant difference in the bleed rates based on technique and device [1]. In that study 7024 children were evaluated, of which a total of 1.4% (n = 99) of children experienced a postoperative hemorrhage that required a second surgery. There was no significant difference found between the four different techniques that were evaluated. Although the combined rates of post-operative hemorrhage requiring a second surgery was lower compared to many other published reports, we hypothesized that continued improvement in surgical techniques could eliminate post-operative hemorrhage completely. As follow up to that study, we decided on a collective change in technique, and subsequent analysis of our post-operative results with respect to only one outcome, post-operative hemorrhage requiring a second surgery. With a new standardized technique in place, we retrospectively evaluated one year of surgical outcomes. These procedures were performed using the technique of a modified and complete, intra-capsular Coblator tonsillectomy. Of the 783 tonsillectomies done in 12 months, we found there were 0 patients that had experienced a postoperative hemorrhage requiring a second surgery. Based on our previous study with a rate of 1.4% post-operative hemorrhage we would have expected to have 11 episodes of post-operative hemorrhage requiring a second surgery.


Physical activity is the key determinant of skeletal muscle mitochondrial function in type 2 diabetes.
Conflicting data exist on mitochondrial function and physical activity in type 2 diabetes mellitus (T2DM) development. The aim was to assess mitochondrial function at different stages during T2DM development in combination with physical exercise in longstanding T2DM patients. We performed cross-sectional analysis of skeletal muscle from 12 prediabetic 11 longstanding T2DM male subjects and 12 male controls matched by age and body mass index. One-year intrasubject controlled supervised exercise training intervention was done in longstanding T2DM patients. Extensive ex vivo analyses of mitochondrial quality, quantity, and function were collected and combined with global gene expression analysis and in vivo ATP production capacity after 1 yr of training. Mitochondrial density, complex I activity, and the expression of Krebs cycle and oxidative phosphorylation system-related genes were lower in longstanding T2DM subjects but not in prediabetic subjects compared with controls. This indicated a reduced capacity to generate ATP in longstanding T2DM patients only. Gene expression analysis in prediabetic subjects suggested a switch from carbohydrate toward lipid as an energy source. One year of exercise training raised in vivo skeletal muscle ATP production capacity by 21 ± 2% with an increased trend in mitochondrial density and complex I activity. In addition, expression levels of β-oxidation, Krebs cycle, and oxidative phosphorylation system-related genes were higher after exercise training. Mitochondrial dysfunction is apparent only in inactive longstanding T2DM patients, which suggests that mitochondrial function and insulin resistance do not depend on each other. Prolonged exercise training can, at least partly, reverse the mitochondrial impairments associated with the longstanding diabetic state.


The epidemiology of firearm suicide in the United States.
Little attention has been given to the role of firearms in suicide. In 1998, firearms were the leading method of committing suicide for both men and women, responsible for three times the number of suicides compared to the next leading method. Understanding the epidemiology of firearm suicide will increase awareness of firearm suicide as a major public health problem. Rates of firearm suicide have changed little over the past two decades and have consistently exceeded rates of firearm homicide. The firearm suicide rate among men is approximately six times that of women. While firearm suicide rates are highest among the elderly, the majority (66%) of firearm suicides are among persons under 55 years of age. Firearm suicide rates among women of all ages have dropped modestly, while rates among elderly men have risen considerably. Whites have roughly twice the rate of firearm suicide as do blacks and other race/ethnicity groups. Individual-level empirical studies have consistently indicated that keeping firearms in the home is associated with an increased risk of suicide. For suicide prevention to be effective, the availability and use of firearms in suicides must be addressed.


HB Handa [alpha 90 (FG 2) Lys replaced by Met]: structure and biosynthesis of a new slightly higher oxygen affinity variant.
Hb Handa, an abnormal hemoglobin which isoelectrofocused anodally to Hb A, was found in a 15-year-old Japanese girl and her mother, who were apparently healthy. Clinical and hematological examinations showed no abnormality in individual heterozygous for this mutant gene except for a slight increase of reticulocyte count (2.1-2.4%). Structural studies disclosed an alpha chain anomaly and substitution of alpha 90 (FG 2) Lys replaced by Met. The content of the abnormal hemoglobin in the hemolysate was 17.5-18.3% of the hemoglobin. The isopropanol precipitation test was negative. The purified abnormal hemoglobin showed a slightly higher oxygen affinity than that of Hb A, but Hill's n constant, Bohr effect and organic phosphate effect were within the normal range. Globin chain biosynthesis in reticulocytes indicated that the production rate of the abnormal alpha chain was suppressed to 38% of the normal level.


Challenges to and the future of medication safety in Saudi Arabia: A qualitative study.
Medication safety is a global concern among healthcare providers. However, the challenges to and the future of medication safety in Saudi Arabia have not been explored. We explored the perspectives of healthcare practitioners on current issues about medication safety in hospitals and community settings in Saudi Arabia in order to identify challenges to improving it and explore the future of medication safety practice. A total of 65 physicians, pharmacists, academics and nurses attended a one-day meeting in March 2010, designed especially for the purpose of this study. The participants were divided into nine round-table discussion sessions. Three major themes were explored in these sessions, including: major factors contributing to medication safety problems, challenges to improving medication safety practice, and participants' suggestions for improving medication safety. The round-table discussion sessions were videotaped and transcribed verbatim and analyzed by two independent researchers. The round-table discussions revealed that major factors contributing to medication safety problems included unrestricted public access to medications from various hospitals and community pharmacies, communication gaps between healthcare institutions, limited use of important technologies such as computerized provider order entry, and the lack of medication safety programs in hospitals. Challenges to current medication safety practice identified by participants included underreporting of medication errors and adverse drug reactions, multilingualism and differing backgrounds of healthcare professionals, lack of communication between healthcare providers and patients, and high workloads. Suggestions for improving medication safety practices in Saudi Arabia included continuous education for healthcare professionals and competency assessment focusing on medication safety, development of a culture that encourages medication error and adverse drug reactions reporting, use of technology proven to decrease medication errors, and promotion and implementation of national patient safety initiatives. Healthcare professionals have identified major challenges and opportunities for medication safety in Saudi Arabia. Policy makers and practitioners should consider these factors when designing future programs aimed at improving the safe use of medications.


Pro-Inflammatory Alterations of Circulating Monocytes in Latent Tuberculosis Infection.
Latent tuberculosis infection (LTBI) has been associated with increased cardiovascular risk. We investigated the activation and pro-inflammatory profile of monocytes in individuals with LTBI and their association with coronary artery disease (CAD). Individuals 40-70 years old in Lima, Peru, underwent QuantiFERON-TB testing to define LTBI, completed a coronary computed tomography angiography to evaluate CAD, and provided blood for monocyte profiling using flow cytometry. Cells were stimulated with lipopolysaccharide to assess interleukin-6 (IL-6) and tumor necrosis factor (TNF)-α responses. The clinical characteristics of the LTBI (n = 28) and non-LTBI (n = 41) groups were similar. All monocyte subsets from LTBI individuals exhibited higher mean fluorescence intensity (MFI) of CX3CR1 and CD36 compared with non-LTBI individuals. LTBI individuals had an increased proportion of nonclassical monocytes expressing IL-6 (44.9 vs 26.9; P = .014), TNF-α (62.3 vs 35.1; P = .014), and TNF-α+IL-6+ (43.2 vs 36.6; P = .042). Among LTBI individuals, CAD was associated with lower CX3CR1 MFI on classical monocytes and lower CD36 MFI across all monocyte subsets. In multivariable analyses, lower CD36 MFI on total monocytes (b = -0.17; P = .002) and all subsets remained independently associated with CAD in LTBI. Individuals with LTBI have distinct monocyte alterations suggestive of an exacerbated inflammatory response and tissue migration. Whether these alterations contribute to cardiovascular disease pathogenesis warrants further investigation.


External Validation of the DynPG for Kidney Transplant Recipients.
In kidney transplantation, dynamic prediction of patient and kidney graft survival (DynPG) may help to promote therapeutic alliance by delivering personalized evidence-based information about long-term graft survival for kidney transplant recipients. The objective of the current study is to externally validate the DynPG. Based on 6 baseline variables, the DynPG can be updated with any new serum creatinine measure available during the follow-up. From an external validation sample of 1637 kidney recipients with a functioning graft at 1-year posttransplantation from 2 European transplantation centers, we assessed the prognostic performance of the DynPG. As one can expect from an external validation sample, differences in several recipient, donor, and transplantation characteristics compared with the learning sample were observed. Patients were mainly transplanted from deceased donors (91.6% versus 84.8%; P < 0.01), were less immunized against HLA class I (18.4% versus 32.7%; P < 0.01) and presented less comorbidities (62.2% for hypertension versus 82.7%, P < 0.01; 25.1% for cardiovascular disease versus 33.9%, P < 0.01). Despite these noteworthy differences, the area under the ROC curve varied from 0.70 (95% confidence interval [CI], 0.64-0.76) to 0.76 (95% CI, 0.64-0.88) for prediction times at 1 and 6 years posttransplantation respectively, and calibration plots revealed reasonably accurate predictions. We validated the prognostic capacities of the DynPG in terms of both discrimination and calibration. Our study showed the robustness of the DynPG for informing both the patient and the physician, and its transportability for a cohort presenting different features than the one used for the DynPG development.


Von Willebrand factor, red cell fragmentation, and disease activity in systemic lupus erythematosus.
This study sought to determine whether the plasma levels of Von Willebrand factor (vWf) and the degree of red blood cell (RBC) fragmentation on peripheral smear correlate with disease activity in systemic lupus erythematosus (SLE). Forty consecutive patients who fulfilled the criteria for SLE were studied prospectively for 1 year. Patients were categorized according to the SLE Disease Activity Index (SLEDAI) as either active (>2) or inactive disease and followed up monthly (active) or quarterly (inactive). At each visit, patients were examined fully and had complete blood count, tests on antibodies to double-stranded DNA, C3, and C4 levels, and urinalysis. Citrated plasma was analyzed for vWf antigen by standard enzyme-linked immunosorbent assay. A Wright's stained blood smear was obtained and schistocytes were quantitated on blood smear. The number of schistocytes per 500 RBCs was determined and a schistocyte index (SI) was calculated. At baseline, vWf correlated with SLEDAI (r = 0.64, p < 0.01), SI correlated with SLEDAI (r = 0.62, p < 0.01), and vWf and SI correlated with each other (r = 0.41, p = 0.01). There was an inverse correlation between baseline C3 levels and vWf (r = 0.49, p = 0.0013) and C3 levels and SI (r = 0.40, p = 0.01). Over time, there was also a correlation of SLEDAI with vWf (r = 0.53, p = 0.002) and SI (r = 0.57;p = 0.002). The relation of vWf with SI approached but did not reach statistical significance (r = 0.37, p = 0.06). We found that the plasma levels of vWf and the degree of RBC fragmentation correlate with lupus disease activity over time. Therefore, inflammation in SLE may be associated with endothelial injury.


Structure of a GPCR ligand in its receptor-bound state: leukotriene B4 adopts a highly constrained conformation when associated to human BLT2.
G protein-coupled receptors (GPCRs) are key players in signal recognition and cell communication and are among the most important targets for drug development. Direct structural information on the conformation of GPCR ligands bound to their receptors is scarce. Using a leukotriene receptor, BLT2, expressed under a perdeuterated form in Escherichia coli , purified in milligram amounts, and folded to its native state using amphipols, we have solved, by (1)H NMR, the structure of receptor-bound leukotriene B4 (LTB4). Upon binding, LTB4 adopts a highly constrained seahorse conformation, at variance with the free state, where it explores a wide range of conformations. This structure provides an experimentally determined template of a pro-inflammatory compound for further pharmacological studies. The novel approach used for its determination could prove powerful to investigate ligand binding to GPCRs and membrane proteins in general.


Outlining a Population "at Risk" of Parkinson's Disease: Evidence from a Case-Control Study.
The multifactorial pathogenesis of Parkinson's Disease (PD) requires a careful identification of populations "at risk" of developing the disease. In this case-control study we analyzed a large Italian population, in an attempt to outline general criteria to define a population "at risk" of PD. We enrolled 300 PD patients and 300 controls, gender and age matched, from the same urban geographical area. All subjects were interviewed on demographics, family history of PD, occupational and environmental toxicants exposure, smoking status, and alcohol consumption. A sample of 65 patients and 65 controls also underwent serum dosing of iron, copper, mercury, and manganese by means of Inductively Coupled-Plasma-Mass-Spectrometry (ICP-MS). Positive family history, toxicants exposure, non-current-smoker, and alcohol nonconsumer status occurred as significant risk factors in our population. The number of concurring risk factors overlapping in the same subject impressively increased the overall risk. No significant differences were measured in the metal serum levels. Our findings indicate that combination of three to four concurrent PD-risk factors defines a condition "at risk" of PD. A simple stratification, based on these questionnaires, might be of help in identifying subjects suitable for neuroprotective strategies.


Tobacco attributable deaths in South Africa.
In mid 1998, a question "Was the deceased a smoker five years ago?" was introduced on the newly revised South African death notification form. A total of 16,230 new death notification forms from 1998 have been coded, and comparison of the prevalence of smoking among those who died of different causes was used to estimate, by case-control comparisons, tobacco attributed mortality in South Africa. Cases comprised deaths from causes known (from other studies) to be causally associated with smoking, and controls comprised deaths from medical conditions expected to be unrelated to smoking. Those who died from external causes, and from diseases strongly related to alcohol consumption, were excluded. Reports were available from 5340 deceased adults (age 25+), whose smoking status was given by a family member. Significantly increased risks were found for deaths from tuberculosis (odds ratio (OR) 1.61, 95% confidence interval (CI) 1.23 to 2.11), chronic obstructive pulmonary disease (COPD) (OR 2.5, 95% CI 1.9 to 3.4), lung cancer (OR 4.8, 95% CI 2.9 to 8.0), other upper aerodigestive cancer (OR 3.0, 95% CI 1.9 to 4.9) and ischaemic heart disease (OR 1.7, 95% CI 1.2 to 2.3). If smokers had the same death rate as non-smokers, 58% of lung cancer deaths, 37% of COPD deaths, 20% of tuberculosis deaths, and 23% of vascular deaths would have been avoided. About 8% of all adult deaths in South Africa (more than 20 000 deaths a year) were caused by smoking.


Negative mood induction: Affective reactivity in recurrent, but not persistent depression.
Despite the high clinical and epidemiological relevance of persistent depression, little is known about its specific psychopathology and whether it is distinct from recurrent depression. Depression in general has been associated with blunted affective reactivity but the evidence from previous studies is inconsistent. Here, we asked whether affective reactivity might differ between persistent and recurrent depression. Twenty patients with persistent depression, 20 patients with recurrent depression and 20 healthy controls (HC) were recruited. Both patient groups showed moderate symptom severity. All participants underwent a sad mood induction procedure. Affective reactivity was assessed with the Positive and Negative Affect Schedule (PANAS) before and after mood induction. We found a striking difference in affective reactivity between patient groups. While the persistent group showed blunted reactivity to mood induction, the recurrent group demonstrated an affective response that was comparable to HC, with an increase in negative and a decrease in positive affect. Blunted affective reactivity was thus specifically associated with persistent in contrast to recurrent depression. These results highlight affective reactivity as an important psychopathological feature that differs between the two patient groups. Preserved affective reactivity to emotional stimuli in the recurrent group might reflect a resilience factor against persistence of depression.


Use of re-esterified palm oils, differing in their acylglycerol structure, in weaning-piglet diets.
Re-esterified oils are new fat sources obtained from chemical esterification of acid oils with glycerol (both economically interesting by-products from oil refining and biodiesel industries, respectively). The different fatty acid (FA) positional distribution and acylglycerol composition of re-esterified oils may enhance the apparent absorption of saturated fatty acids (SFA) and, thus, their overall nutritive value. The aim of the present study was to investigate the potential use of re-esterified palm oils, in comparison with their corresponding acid and native oils, and also with an unsaturated fat source in weaning-piglet diets. The parameters assessed were: FA apparent absorption, acylglycerol and free fatty acid (FFA) composition of feces, and growth performance. One-hundred and twenty weaning piglets (average weight of 8.50±1.778 kg) were blocked by initial BW (six blocks) and randomly assigned to five dietary treatments, resulting in four piglets per pen and six replicates per treatment. Dietary treatments were a basal diet supplemented with 10% (as-fed basis) of native soybean oil (SN), native palm oil (PN), acid palm oil (PA), re-esterified palm oil low in mono- (MAG) and diacylglycerols (DAG) (PEL), or re-esterified palm oil high in MAG and DAG (PEH). Results from the digestibility balance showed that SN reached the greatest total FA apparent absorption, and statistically different from PN, PA and PEL (P0.05), but PEH achieved the greatest total FA apparent absorption. Animals fed PEL, despite the fact that PEL oil contained more sn-2 SFA, did not show an improved absorption of SFA (P>0.05). Animals fed PA and PN showed similar apparent absorption coefficients (P>0.05), despite the high FFA content of PA oil. The acylglycerol and FFA composition of feces was mainly composed of FFA. There were no significant differences in growth performance (P>0.05). Results of the present study suggest that, despite the different acylglycerol structure of re-esterified oils, there were no significant differences in digestibility or performance with respect to their corresponding PN and PA oils in weaning-piglet diets.


Cumulative evidence of relationships between multiple variants in 8q24 region and cancer incidence.
Genome-wide association studies (GWAS) have identified multiple independent cancer susceptibility loci at chromosome 8q24. We aimed to evaluate the associations between variants in the 8q24 region and cancer susceptibility. A comprehensive research synopsis and meta-analysis was performed to evaluate associations between 28 variants in 8q24 and risk of 7 cancers using data from 103 eligible articles totaling 146,932 cancer cases and 219,724 controls. Results: 20 variants were significantly associated with risk of prostate cancer, colorectal cancer, thyroid cancer, breast cancer, bladder cancer, stomach cancer, and glioma, including 1 variant associated with prostate cancer, colorectal cancer, and thyroid cancer. Cumulative epidemiological evidence of an association was graded as strong for DG8S737 -8 allele, rs10090154, rs7000448 in prostate cancer, rs10808556 in colorectal cancer, rs55705857 in gliomas, rs9642880 in bladder cancer, moderate for rs16901979, rs1447295, rs6983267, rs7017300, rs7837688, rs1016343, rs620861, rs10086908 associated in prostate cancer, rs10505477, rs6983267 in colorectal cancer, rs6983267 in thyroid cancer, rs13281615 in breast cancer, and rs1447295 in stomach cancer, weak for rs6983561, rs13254738, rs7008482, rs4242384 in prostate cancer. Data from ENCODE suggested that these variants with strong evidence and other correlated variants might fall within putative functional regions. Our study provides summary evidence that common variants in the 8q24 are associated with risk of multiple cancers in this large-scale research synopsis and meta-analysis. Further studies are needed to explore the mechanisms underlying variants in the 8q24 involved in various human cancers.


Strontium isotope evidence for Neanderthal and modern human mobility at the upper and middle palaeolithic site of Fumane Cave (Italy).
To investigate the mobility patterns of Neanderthals and modern humans in Europe during the Middle-to-Upper Palaeolithic transition period, we applied strontium isotope analysis to Neanderthal (n = 3) and modern human (n = 2) teeth recovered from the site of Fumane Cave in the Monti Lessini region of Northern Italy. We also measured a large number of environmental samples from the region, to establish a strontium 'baseline', and also micromammals (vole teeth) from the levels associated with the hominin teeth. We found that the modern humans and Neanderthals had similar strontium isotope values, and these values match the local baseline values we obtained for the site and the surrounding region. We conclude that both groups were utilizing the local mountainous region where Fumane Cave is situated, and likely the nearby Lessini highlands and Adige plains, and therefore the strontium evidence does not show differening mobility patterns between Neanderthals and modern humans at the Fumane site.


Thromboembolic meningoencephalitis diagnosed in Natal.
Nine feedlot cattle showed clinical signs consistent with those expected in thromboembolic meningoencephalitis. These signs included pyrexia, ataxia, posterior paresis, paralysis and coma. Brown necrotic foci with haemorrhagic borders were observed in the brains of three animals that had died. In these foci vasculitis, thrombosis, infarction and neutrophil infiltration were observed during microscopical examination. Haemophilus somnus was isolated in pure culture from the brains.


[Follow-up survey of dioxins in the blood of Yusho patients (in 2001)].
Follow-up survey of the blood concentration of polychlorinated dibezo-p-dioxin (PCDDs), polychlorinated dibenzofuran (PCDFs), Non-ortho-coplaner PCBs (Non-Co-PCB), and Mono-ortho-coplaner PCBs (Mo-Co-PCB) in Yusho patients is very important for their health control. We determined the blood concentration of these dioxin-like isomers in 78 blood samples collected in 2001 using by a high-resolution gas chromatograph/high-resolution mass spectrometer (HRGC/HRMS) equipped with a solvent-cut large volume injection system (SCLV). The accelerated solvent extraction (ASE) method was employed for the treatment of blood samples. The method employed here can allow a reduced amount of blood collecting from Yusho patients compared with the conventional method. In typical Yusho patients (Group A of PCB pattern), the mean TEQ concentrations of PCDDs, PCDFs, Non-Co-PCBs, and Mono-Co-PCBs in blood collected in 2001 were 36, 260, 9.3, and 64 pg-TEQ/g lipid, respectively. As well, the toxic contribution of PCDDs, PCDFs, Non-Co-PCBs, and Mono-Co-PCBs in typical Yusho patients (Group A of PCB pattern) were 10, 70, 3, and 17% of the total TEQ value, respectively. The toxic contribution rate of PCDFs TEQ was still at high levels in relation to total TEQ in typical Yusho patients as mention above. The Yusho patients showing B pattern also had the same A pattern contribution rate. In the patients of group C, however, the mean blood levels of dioxin TEQ in 2001 was two times those of the normal controls determined in 2000 in Fukuoka prefecture. As well, the contribution rates were similar to those of normal subjects, although some patients in this group showed a high toxic contribution rate of PCDFs TEQ in relation to total TEQ. Thirty-three years had passed since the Yusho accident, and the total blood concentrations of PCDFs in the blood of Yusho patients were still higher than those of normal subjects. Among the different groups of PCB patterns, the concentration of 2,3,4,7,8-PeCDF, 1,2,3,4,7,8-HxCDF and 1,2,3,6,7,8-HxCDF showed significant differences. These findings indicate that these isomers are not metabolized in the human body since traces of them continued to be detected in these patients since they had been contaminated by the rice oil in the original incident.


Alcohol consumption patterns among young people from rural areas of Lublin province.
The article aims at the increasing of knowledge among young people coming from the country, studying at various types of schools--from the aspect of frequency of their drinking, age of alcohol initiation and preferences towards alcoholic drinks. We examined 5,000 young people from rural areas in Lublin province between the ears 2002-2004. A screening questionnaire that included AUDIT test (the Alcohol Use Disorders Identification Test) was used. The AUDIT was developed by the WHO to identify hazardous and harmful alcohol consumption. The results showed that both frequency and alcohol initiation age were related to type of school and sex of young people. We found that young people from rural areas of Lublin province do not differ from all-Polish trends in relation to these patterns. Some changes in the alcohol consumption model in Polish villagers were noted. Young people in the country clearly prefer beer drinks. Restrictions regarding alcohol sales to young people are not complied with. In almost half of the group we found alcoholism in families. Our findings show that antialcoholic prophylaxis should be initiated in the first years of childrens' education. Preventive programmes should be based not only on school, but also on the family.


Reversal of a cholinergic-induced deficit in a rodent model of recognition memory by the selective 5-HT6 receptor antagonist, Ro 04-6790.
Accumulating evidence suggests a potential role for the 5-HT(6 )receptor in cognitive function and the potential use of 5-HT(6) receptor antagonists in the treatment of learning and memory disorders. The aim of the current study was to investigate the effect of the selective 5-HT(6) receptor antagonist, Ro 04-6790, on both the performance of normal adult rats and restoration of a pharmacological disruption of memory function produced by the non-selective muscarinic receptor antagonist, scopolamine, or the dopamine D(2) receptor antagonist, raclopride, in a rodent model of recognition memory. Passive, perceptually based, recognition memory was assessed using a novel object discrimination task. Following habituation to an arena, rats were presented with two identical objects during trial 1 (T(1)) and a novel and familiar object during trial 2 (T(2)). The time spent exploring the two objects in each trial was measured and novel object discrimination assessed in T(2). In the absence of drug all rats spent an equal time exploring the two identical objects in T(1) but more time exploring the novel object in T(2). Scopolamine (but not N-methylscopolamine) and raclopride both produced a dose-dependent reduction in novel object discrimination whilst the 5-HT(6) receptor antagonist, Ro 04-6790, had no effect on discrimination when given alone but completely reversed the scopolamine- but not the raclopride-induced deficit. This study demonstrates that acute administration of Ro 04-6790 reverses a cholinergic but not a dopaminergic deficit in a rodent model of recognition memory and provides further support for a role of the 5-HT(6) receptor in the regulation of cognitive function.


[Extrarenal variant of an embryonal nephroma].
A rare variant of embryonal nephroma in a man of 18 is described. Epithelial and mesodermal components were markedly distinctive in the tumour. The epithelial part of the tumour was represented by tubules resembling renal tubules, and glomerulo-like structures, and the mesodermal part consisted of cross-striated muscles, edematous connective tissue and osteoid substance.


Identifying pathogenic genetic background of simplex or multiplex retinitis pigmentosa patients: a large scale mutation screening study.
More than half of the retinitis pigmentosa (RP) cases are genetically simplex or multiplex. To date, 37 causative genes of RP have been identified; however, the elucidation of gene defects in simplex or multiplex RP patients/families remains problematic. The aim of our study was to identify the genetic causes of RP in patients with unknown or non-Mendelian inheritance. Since 2003, 52 simplex RP patients, 151 patients from 141 multiplex RP families, and six sporadic patients with retinal degeneration were studied. A total of 108 exons of 30 RP-causing genes that harboured the reported mutations were screened by an efficient denaturing high performance liquid chromatography (dHPLC) based assay. Aberrant fragments were subsequently analysed by automatic sequencing. Twenty-six mutations, including two frameshift mutations, one single amino acid deletion, and 23 missense mutations, were identified in 28 probands (14.07%). Eighteen mutations have not been reported to date. Three pairs of combined mutations in different genes were identified in two sporadic cases and one multiplex family, indicating the possibility of novel digenic patterns. Of the 23 missense mutations, 21 were predicted as deleterious mutations by computational methods using PolyPhen, SIFT, PANTHER, and PMut programs. We elucidated the mutation spectrum in Japanese RP patients and demonstrated the validity of the mutation detection system using dHPLC sequencing for genetic diagnosis in RP patients independent of familial incidence, which may provide a model strategy for identifying genetic causes in other diseases linked to a wide range of genes.


Nicotine exposure and decontamination on tobacco harvesters' hands.
Green tobacco sickness is an illness associated with nicotine exposures among tobacco harvesters. Agricultural workers manually harvest tobacco and thus have the potential for skin exposure to nicotine, particularly on the hands. Often gloves are not worn as it hinders the harvesters' ability to harvest the tobacco leaves. The purposes of this study were to measure the concentration of nicotine residue on the hands of tobacco harvesters and the effectiveness of hand washing at removing the residue. Wipe samples from the hands of 12 tobacco harvesters were collected at the end of morning and afternoon work periods over two consecutive days. Each harvester had one hand wiped before washing his hands, and the other hand wiped after washing his hands with soap and water. Eight samples per worker were collected over the two days for a total of 96 samples collected. In addition to the hand-wipe samples, leaf-wipe samples were collected from 15 tobacco plants to estimate the amount of nicotine residue on the plants. The average nicotine level in leaf-wipe samples was 1.0 microg cm(-2). The geometric mean pre-wash and post-wash nicotine levels on the hands were 10 and 0.38 microg cm(-2), respectively. Nicotine leaf-wipe level, right or left hand and time of sampling did not significantly influence exposure. Job position-working on the bottom versus the top of the tobacco harvesting machine-was associated with nicotine levels. Pre-wash nicotine levels were higher for workers on the bottom of the harvester but not significantly higher (P = 0.17). Post-wash nicotine levels were significantly higher for workers on the bottom of the harvester (P = 0.012). A substantial amount of nicotine was transferred to the hands, but washing with soap and water in the field significantly reduced nicotine levels by an average of 96% (P < 0.0001).


Tetrachloroethene conversion to ethene by a Dehalococcoides-containing enrichment culture from Bitterfeld.
A Dehalococcoides-dominated culture coupling reductive dechlorination of tetrachloroethene (PCE) to ethene to growth was enriched from a European field site for the first time. Microcosms were set up using groundwater from a chlorinated ethene-contaminated anaerobic aquifer in Bitterfeld (Germany). Active, lactate-amended microcosms capable of PCE dechlorination to ethene without the accumulation of intermediates were used for further enrichment. After three transfers on lactate as an electron donor and PCE as an electron acceptor, the enrichment was transferred to parallel cultures with one of the chlorinated ethenes as an electron acceptor and acetate and hydrogen as the carbon and energy source, respectively. After three more transfers, a highly purified culture was derived that was capable of dechlorinating PCE with hydrogen and acetate as the electron donor and carbon source, respectively. PCR, followed by denaturing gradient gel electrophoresis, cloning and sequencing revealed that this culture was dominated by a Dehalococcoides sp. belonging to the Pinellas group. Investigation of substrate specificity in the parallel cultures suggested the presence of a novel Dehalococcoides that can couple all dechlorination steps, from PCE to ethene, to energy conservation. Quantitative real-time PCR confirmed growth with PCE, cis-dichloroethene, 1,1-dichloroethene or vinyl chloride as electron acceptors. The culture was designated BTF08 due to its origin in Bitterfeld.


Development of a nutritional screening and assessment indicator system for patients with esophageal cancer in China: Findings from the Delphi method.
In China, individuals diagnosed with esophageal cancer are confronted with an elevated risk of nutritional inadequacy or malnutrition throughout the course of their disease, a condition that contributes to various adverse clinical outcomes. A vast corpus of data are burgeoning at an unprecedented rate, primarily due to the revolutionary growth of digitalization technologies and artificial intelligence, notably within the domains of health care and medicine. The purpose of this investigation is to initiate the development of a nutritional screening and assessment indicator framework for patients with esophageal cancer within the Chinese context. We seek to furnish an instrumental reference to facilitate preparations for the forthcoming era of advanced, "deep," evidence-based medicine. An integrative methodology was employed to forge the preliminary draft of the nutritional screening and assessment indicator system for preoperative patients with esophageal cancer. This encompassed a rigorous literature survey, in-depth clinical practice investigation, and the facilitation of expert panel discussions. Thereafter, two iterative consultation phases were conducted using the Delphi method in China. The analytic hierarchy process was deployed to ascertain the weighting of each index within the definitive evaluation indicator system. The effective response rates for the dual rounds of expert consultation were 91.7% and 86.4%, with commensurate authority coefficients of 0.97 and 0.91. The Kendall harmony coefficients were ascertained to be 0.19 and 0.14 (p < 0.01), respectively. The culminating nutritional screening and assessment indicator system for patients with esophageal cancer comprised 5 primary-level indicators and 38 secondary-level indicators. The nutritional screening and assessment indicator system contrived for patients with esophageal cancer is underpinned by cogent theoretical principles, leverages an astute research methodology, and manifests dependable outcomes. This system may be appositely utilized as a meaningful reference for the nutritional screening and assessment process in patients afflicted with esophageal cancer.


Mental healthcare staff's knowledge and experiences of diabetes care for persons with psychosis--a qualitative interview study.
This paper aims to explore and analyse mental healthcare staff's (MHCS) knowledge and experiences of diabetes care for persons with psychosis. There are a range of studies concerning the increased risk of type 2 diabetes mellitus among persons with psychosis, and the need for healthy lifestyle interventions to prevent the illness. MHCS are often trusted and have regular follow-ups with the patients, and their attitudes and actions often play an important role for the person's care behaviour. There is still little documentation of their experiences of diabetes care. A qualitative, explorative design was used, collecting data through semi-structured interviews with 12 MHCS working in psychosis outpatient care in Sweden. Data were analysed with qualitative content analysis. Findings Three categories emerged and provide a deeper understanding of how staff were aware of the risks of type 2 diabetes among their patients and therefore performed lifestyle interventions to promote these. Nevertheless, they lacked knowledge of diabetes care and simultaneously felt a lack of training among diabetes nurses to adapt diabetes care to suit persons with cognitive dysfunctions. Patients who were overconfident in their ability to manage diabetes care reported to have experienced most difficulties. Cooperation among those involved in these persons' health was considered necessary. Implications Diabetes care for persons with psychosis could improve if knowledge of type 2 diabetes was increased among MHCS and training in how to adapt diabetes care to persons with cognitive dysfunctions was enlarged among diabetes nurses. A challenge for nurses is to see how the care of different illnesses and support given by the family and others affect the persons total life situation and health. Healthcare plans and cooperation among all those involved in these persons' health is necessary for this.


Structural basis for ion selectivity in TMEM175 K+ channels.
The TMEM175 family constitutes recently discovered K+channels that are important for autophagosome turnover and lysosomal pH regulation and are associated with the early onset of Parkinson Disease. TMEM175 channels lack a P-loop selectivity filter, a hallmark of all known K+ channels, raising the question how selectivity is achieved. Here, we report the X-ray structure of a closed bacterial TMEM175 channel in complex with a nanobody fusion-protein disclosing bound K+ ions. Our analysis revealed that a highly conserved layer of threonine residues in the pore conveys a basal K+ selectivity. An additional layer comprising two serines in human TMEM175 increases selectivity further and renders this channel sensitive to 4-aminopyridine and Zn2+. Our findings suggest that large hydrophobic side chains occlude the pore, forming a physical gate, and that channel opening by iris-like motions simultaneously relocates the gate and exposes the otherwise concealed selectivity filter to the pore lumen.


Gastrointestinal and metabolic effects of amylase inhibition in diabetics.
A partially purified amylase inhibitor given with a single meal causes maldigestion of carbohydrate, increases delivery of carbohydrate to the ileum, and reduces postprandial plasma glucose. To determine the effect of more prolonged administration of the inhibitor on gastrointestinal function and carbohydrate tolerance, we studied 6 non-insulin-dependent diabetics (3 previously treated with oral agents and 3 treated with diet alone) for 3 wk while they ate a weight-maintenance diet. Patients taking oral agents continued them during the first week. During the second week, 4-6 g of the inhibitor was given with each meal. Capillary blood glucose concentration was measured before each meal and 90 min postprandially. On the last day of each week venous blood samples for glucose, hormones, and lactic acid analysis and a quantitative stool culture were obtained. Total carbohydrate absorption was estimated by comparing postprandial breath hydrogen on study days 7, 14, and 21 with breath hydrogen after ingesting 15 g of lactulose on days 0, 15, and 22. There 24-h stools were collected and weighed at the end of each week and analyzed for carbohydrate, lactic acid, short-chain fatty acids, pH, dry matter, amylase, and fat. The inhibitor significantly (p less than 0.05) reduced postprandial plasma glucose, C-peptide, insulin, and gastric inhibitory polypeptide concentrations, significantly increased (p less than 0.05) breath hydrogen excretion, and caused carbohydrate malabsorption. Diarrhea occurred the first day the inhibitor was ingested, but thereafter cessation of diarrhea was associated with changes in the metabolism of carbohydrate by colonic flora. As the amylase inhibitor improves carbohydrate homeostasis and is not associated with continuing diarrhea, it may be a useful adjuvant in the treatment of patients with non-insulin-dependent diabetes mellitus.


KIF13B-NRG1 Gene Fusion and KRAS Amplification in a Case of Natural Progression of Lung Cancer.
An 85-year-old woman with a history of several primary lung cancers presented with liver metastases. The primary lung cancers were all managed surgically and the patient did not receive adjuvant or neoadjuvant therapy prior to presenting with metastases. Comparison of molecular testing results from the most recent primary and the liver metastases demonstrated ( a) a clonal relationship between the 2 cancers and ( 2) the presence of a KIF13B-NRG1 fusion and KRAS amplification unique to the metastases. When integrated, the molecular surgical pathology findings in this case illustrate the extent of "oncogenic drive" in preterminal lung cancer.


The Oncogene Metadherin Interacts with the Known Splicing Proteins YTHDC1, Sam68 and T-STAR and Plays a Novel Role in Alternative mRNA Splicing.
Oncogenic metadherin is a key contributor to tumourigenesis with metadherin expression and cytoplasmic localisation previously linked to poor survival. A number of reports have shown metadherin localises specifically to nuclear speckles known to be rich in RNA-binding proteins including the splicing proteins YTHDC1, Sam68 and T-STAR, that have been shown to select alternative splice sites in mRNA of tumour-associated proteins including BRCA, MDM2 and VEGF. Here we investigate the interaction and relationship between metadherin and the splice factors YTHDC1, T-STAR and Sam68. Using a yeast two-hybrid assay and immunoprecipitation we show that metadherin interacts with YTHDC1, Sam68 and T-STAR and demonstrate that T-STAR is significantly overexpressed in prostate cancer tissue compared to benign prostate tissue. We also demonstrate that metadherin influences splice site selection in a dose-dependent manner in CD44v5-luc minigene reporter assays. Finally, we demonstrate that prostate cancer patients with higher metadherin expression have greater expression of the CD44v5 exon. CD44v5 expression could be used to discriminate patients with poor outcomes following radical prostatectomy. In this work we show for the first time that metadherin interacts with, and modulates, the function of key components of splicing associated with cancer development and progression.


Performance of Chromogenic Candida agar and CHROMagar Candida in recovery and presumptive identification of monofungal and polyfungal vaginal isolates.
Chromogenic Candida agar (OCCA) is a novel medium facilitating isolation and identification of Candida albicans, C. tropicalis, and C. krusei, as well as indicating polyfungal population in clinical samples. We compare the performance of OCCA, to CHROMagar Candida (CAC) and Sabouraud chloramphenicol agar (SCA). Vaginal swab samples from 392 women were simultaneously inoculated onto three study media. A total of 161 (41.1%) were found to be positive for fungi of which 140 (87%) were monofungal, and 21 (13%) polyfungal. One-hundred and fifty-seven samples (97.5%) were positive on CAC, 156 (96.9%) on OCCA, 148 (91.9%) on SCA and 144 (89.4%) samples were positive on all three media. The yeasts were identified by conventional methods including germ tube test, microscopic morphology on cornmeal-Tween 80 agar, and the commercial API 20C AUX. The 182 isolates were C. albicans (n = 104), C. glabrata (n = 51), C. krusei (n = 7), C. tropicalis (n = 5), C. famata (n = 3), C. kefyr (n = 3), C. zeylanoides (n = 3), C. colliculosa (n = 2), and other species of Candida (n = 4). Among the 21 polyfungal populations, 20 (95.2%) were detected in OCCA, 14 (66.7%) in CAC, and 13 (61.9%) in CAC and OCCA (P <0.05). Most polyfungal populations (47.6%) yielded C. albicans + C. glabrata. The efficiency of both chromogenic media for C. albicans was >or=92.9% at 72 h. OCCA is more efficient and reliable for rapidly identifying C. albicans and polyfungal populations than CAC. However, CAC is more efficient for identifying C. krusei and C. tropicalis. A chromogenic agar with a higher isolation rate of yeasts and better detection of polyfungal populations than SCA, is suggested as a medium of first choice when available.


Allopurinol intake does not modify the slow component of V(.)O(2) kinetics and oxidative stress induced by severe intensity exercise.
The aim of this study was to test the hypothesis that allopurinol ingestion modifies the slow component of V(.)O(2) kinetics and changes plasma oxidative stress markers during severe intensity exercise. Six recreationally active male subjects were randomly assigned to receive a single dose of allopurinol (300 mg) or a placebo in a double-blind, placebo-controlled crossover design, with at least 7 days washout period between the two conditions. Two hours following allopurinol or placebo intake, subjects completed a 6-min bout of cycle exercise with the power output corresponding to 75 % V(.)O(2)max. Blood samples were taken prior to commencing the exercise and then 5 minutes upon completion. Allopurinol intake caused increase in resting xanthine and hypoxanthine plasma concentrations, however it did not affect the slow component of oxygen uptake during exercise. Exercise elevated plasma inosine, hypoxanthine, and xanthine. Moreover, exercise induced a decrease in total antioxidant status, and sulfhydryl groups. However, no interaction treatment x time has been observed. Short term severe intensity exercise induces oxidative stress, but xanthine oxidase inhibition does not modify either the kinetics of oxygen consumption or reactive oxygen species overproduction.


Graph-based Clustering of miRNA Sequences.
The many miRNAs discovered so far have been divided into biologically representative families, aiming at organizing and systematizing their study so to promote, mainly, a better understanding of their functionalities. Clustering miRNA sequences can corroborate the family-based organizations as well as helping to explore sequences belonging to the same cluster as potentially having similar biological functions. Considering that members of the same miRNA family tend to biologically function in similar ways, a well-structured family can help detecting miRNA functions which have not been associated yet with any existing family. The work described in this paper empirically investigates the suitability of organizing miRNAs as families, using a clustering algorithm based on a particular type of graph i.e., minimal spanning tree (MST), for clustering miRNA sequences. Seven miRNA families stored in the online miRBase database have been used as input to the MST-based clustering algorithm and clustering results have been compared to assess the suitability of identirying them. The motivations for the experiments were to identify refinements in miRNA family organizations that could be pursued and, also, investigate how the chosen graph-based clustering algorithm would perform in miRNA related domains. Interesting and useful results, particularly related to detecting information based on the visualization of the final induced graphs, and their induced connected components (clusters), are presented and discussed. Particularly, experiments results suggested the possibility of refining some miRNA families by grouping some of their miRNAs as sub-families.


Long term improvement of glucose homeostasis by vanadate treatment in diabetic rats.
The trace element vanadium (V) exerts insulin-like effects in vitro. The present study examined its effects on glucose homeostasis in rats made diabetic by streptozotocin. Na3VO4 (0.2 or 0.5 mg/ml) was administered ad libitum in drinking water. Fed plasma glucose levels (approximately 26 mmol/liter) fell by 30% and 56% after 5 days of treatment with the low (VO.2) and high (VO.5) concentrations of vanadate, respectively. This decrease was not due to a rise in peripheral insulin levels and persisted for more than 2 months. Daily glucosuria was decreased by 60% and 85% in VO.2 and VO.5 rats, respectively. Tolerance of the rats to oral or iv glucose was also considerably improved by vanadate; integrated glucose responses were about 55% and 75% lower in VO.2 and VO.5 rats than in controls, and the differences were not due to restoration of insulin release. Compared to nondiabetic rats, pancreatic insulin reserves amounted to 1% in untreated rats, 3% in VO.2 rats, and 6% in VO.5 rats after 9 weeks of treatment. Liver, but not muscle, glycogen was increased by vanadate. Despite improvement of their diabetic state, vanadate-treated rats did not gain more weight than untreated rats. Their food intake (corrected for urinary glucose losses) was decreased by about 25%. No signs of altered kidney or liver function were observed in rats receiving vanadate. In conclusion, vanadate markedly improves glucose homeostasis in streptozotocin-diabetic rats by an insulin-like mechanism, but does not reproduce the anabolic effects of the hormone.


Quantifying the reproducibility of heart position during treatment and corresponding delivered heart dose in voluntary deep inhalation breath hold for left breast cancer patients treated with external beam radiotherapy.
Voluntary deep inhalation breath hold (VDIBH) reduces heart dose during left breast irradiation. We present results of the first study performed to quantify reproducibility of breath hold using bony anatomy, heart position, and heart dose for VDIBH patients at treatment table. Data from 10 left breast cancer patients undergoing VDIBH whole-breast irradiation were analyzed. Two computed tomography (CT) scans, free breathing (FB) and VDIBH, were acquired to compare dose to critical structures. Pretreatment weekly kV orthogonal images and tangential ports were acquired. The displacement difference from spinal cord to sternum across the isocenter between coregistered planning Digitally Reconstructed Radiographs (DRRs) and kV imaging of bony thorax is a measure of breath hold reproducibility. The difference between bony coregistration and heart coregistration was the measured heart shift if the patient is aligned to bony anatomy. Percentage of dose reductions from FB to VDIBH: mean heart dose (48%, SD 19%, p = 0.002), mean LAD dose (43%, SD 19%, p = 0.008), and maximum left anterior descending (LAD) dose (60%, SD 22%, p = 0.008). Average breath hold reproducibility using bony anatomy across the isocenter along the anteroposterior (AP) plane from planning to treatment is 1 (range, 0-3; SD, 1) mm. Average heart shifts with respect to bony anatomy between different breath holds are 2 ± 3 mm inferior, 1 ± 2 mm right, and 1 ± 3 mm posterior. Percentage dose changes from planning to delivery: mean heart dose (7%, SD 6%); mean LAD dose, ((9%, SD 7%)S, and maximum LAD dose, (11%, SD 11%) SD 11%, p = 0.008). We observed excellent three-dimensional bony registration between planning and pretreatment imaging. Reduced delivered dose to heart and LAD is maintained throughout VDIBH treatment.


The effect of maternal position on venous return for pregnant women during MRI.
Magnetic resonance imaging (MRI) in pregnancy is commonly undertaken in the left lateral tilt (LLT) position to prevent inferior vena cava (IVC) compression and supine hypotensive events, although this may be suboptimal for image quality. The supine position may also have an adverse effect on fetal well-being. The spinal venous plexus may provide an alternative pathway for venous return in the presence of IVC compression. This study assesses morphology and blood flow of the IVC and spinal venous plexus for pregnant women in LLT and supine positions to ascertain the effect of maternal position on venous return during MRI. Eighty-two pregnant women underwent phase contrast MRI (PC-MRI) of the IVC and spinal venous plexus in the supine position; 25 were also imaged in the LLT position. Differences in life monitoring, IVC, spinal venous plexus and total venous return between the two positions were assessed. A linear regression assessed the relationship between flow in the IVC and the spinal venous plexus in the supine position. Increasing gestational age and the right-sided position of the uterus on IVC and spinal venous plexus venous return were also evaluated. Hypotension symptoms were similar in supine (10%) and LLT (8%) positioning. Supine positioning decreased IVC height (p < 0.004) and flow (p = 0.045) but flow in the spinal venous plexus increased (p < 0.001) compared with the LLT position. Total venous return showed no difference (p = 0.989) between the two positions. Additional measurements of flow in the aorta also showed no significant difference between the two groups (p = 0.866). Reduced IVC flow in the supine position was associated with increasing gestational age (p = 0.004) and degree of right-sided uterine position (p = 0.004). Women in the left lateral decubitus position who then rotated supine had greater flow in the IVC (p = 0.008) and spinal venous plexus (p = 0.029) than those who started supine. For the majority of women, the spinal venous plexus acts as a complementary venous return system for pregnant women in the supine position, maintaining vascular homeostasis. Further study is needed to assess the effects on the health of the fetus.


In vitro and in silico strategies to identify OATP1B1 inhibitors and predict clinical drug-drug interactions.
To establish in vitro and in silico models that predict clinical drug-drug interactions (DDIs) with the OATP1B1 (SLCO1B1) transporter. The inhibitory effect of 146 drugs and drug-like compounds on OATP1B1-mediated transport was studied in HEK293 cells. A computational model was developed to predict OATP1B1 inhibition. Concentration-dependent effects were investigated for six compounds; clinical DDIs were predicted by calculating change in exposure (i.e. R-values) in eight different ways. Sixty-five compounds were identified as OATP1B1 inhibitors at 20 μM. The computational model predicted the test set with 80% accuracy for inhibitors and 91% for non-inhibitors. In vitro-in vivo comparisons underscored the importance of using drugs with known clinical effects as references. Thus, reference drugs, cyclosporin A, gemfibrozil, and fenofibrate, provided an inhibition interval to which three antiviral drugs, atazanavir, lopinavir, and amprenavir, could be compared and their clinical DDIs with OATP1B1 classified. Twenty-two new OATP1B1 inhibitors were identified, a predictive OATP1B1 inhibition in silico model was developed, and successful predictions of clinical DDIs were obtained with OATP1B1.


Human diabetic endoneurial sorbitol, fructose, and myo-inositol related to sural nerve morphometry.
Fascicles of the sural nerve from each of 20 diabetic patients, mostly with maturity-onset diabetes, were studied by biochemical and pathological techniques, and results were compared to values found in nerve specimens from 15 healthy persons. The sorbitol and fructose content was much more variable in diabetic than in healthy nerves. More than one-third of the diabetic nerves had sorbitol and fructose values above the highest levels for controls. myo-Inositol and scyllo-inositol content was not reduced in diabetic nerves. The sorbitol, fructose, and inositol concentrations could not be related to clinical, neurophysiological, or pathological severity of neuropathy. A comparison of scored symptoms and signs and clinical neurophysiological studies against morphometric and teased fiber studies of sural nerve demonstrated that the former three provide sensitive and reliable measures of severity of neuropathy that can be used for controlled clinical trials of diabetic neuropathy. The presence and type of teased fiber abnormalities could be related to the duration of diabetes and to symptoms of neuropathy. In untreated diabetics without symptoms of neuropathy, a higher than normal frequency of teased fibers showing segmental demyelination and remyelination was found. Untreated diabetics with symptomatic neuropathy showed two kinds of abnormalities: fibers with segmental demyelination and remyelination and fibers undergoing axonal degeneration. In treated diabetics, who often had longstanding neuropathy, the most common abnormalities were fibers undergoing axonal degeneration.


Impact of diabetes mellitus on clinical outcomes in patients undergoing surgical resection for pancreatic cancer: a retrospective, cohort study.
Diabetes mellitus (DM) has a bidirectional association with pancreatic cancer (PaC); however, its effect on clinical outcomes has not been thoroughly evaluated. We analyzed these data in a large sample of PaC subjects who had undergone surgical resection. Subjects enrolled in the Mayo Clinic Pancreatic Cancer SPORE registry from 2000 to 2010 who had resection with curative intent were identified (n=488). Tumor size, cancer stage, and postoperative median survival were evaluated. Median survivals were compared with Kaplan-Meier curves and Cox proportional hazards regression modeling. A total of 275 (56%) subjects had DM before surgery. DM subjects had larger tumors compared with those without DM (3.6 cm vs. 3.3, P=0.002), even after controlling for covariates including age, body mass index, and tumor grade. Cancer stage at the time of surgery was not affected by DM status (P=0.575). Preoperative DM was not associated with an increased risk of death using a multivariable survival analysis (hazard ratio 1.06, 95% confidence interval 0.81-1.38, P=0.676). The median survival following cancer resection was similar between subjects with and without DM (24 vs. 26 months, P=0.610). In addition, postoperative survival was similar on the basis of the duration of DM (new-onset vs. long-standing) and prior use of antidiabetic treatments in diabetic subjects. PaC subjects with DM have larger tumors than nondiabetic subjects. Despite this observation, preoperative DM does not negatively impact the cancer stage at the time of surgery or postoperative survival. Thus, the effect of DM on tumor size is either overshadowed by early metastatic spread of the cancer or is mitigated by the tumor resection.


Difference in the electromyographic onset of the deep and superficial multifidus during shoulder movement while standing.
Based on the current literature, it remains unclear whether electromyographic onset of the deep fibers of the multifidus (DM) is dependent on the direction of shoulder movement and the position of the center of foot pressure (CFP). In the present study, we re-examined the electromyographic onset of the DM during shoulder flexion and extension and investigated the influence of the CFP position before arm movement. Intramuscular and surface electrodes recorded the electromyographic onset of the DM, superficial fibers of the multifidus (SM), rectus abdominis, and anterior and posterior deltoid. Eleven healthy participants performed rapid, unilateral shoulder flexion and extension in response to audio stimuli at three CFP positions: quiet standing, extreme forward leaning, and extreme backward leaning. It was found that the electromyographic onset of the DM and SM relative to the deltoid was dependent on the direction of arm movement. Additionally, of all electromyographic onsets recorded, only that of the DM occurred earlier in the extreme forward leaning position than in the extreme backward leaning position during shoulder flexion. These results suggest that the electromyographic onset of DM was influenced by the biomechanical disturbance such as shoulder movement and CFP position.


Models with indirect genetic effects depending on group sizes: a simulation study assessing the precision of the estimates of the dilution parameter.
In settings with social interactions, the phenotype of an individual is affected by the direct genetic effect (DGE) of the individual itself and by indirect genetic effects (IGE) of its group mates. In the presence of IGE, heritable variance and response to selection depend on size of the interaction group (group size), which can be modelled via a 'dilution' parameter (d) that measures the magnitude of IGE as a function of group size. However, little is known about the estimability of d and the precision of its estimate. Our aim was to investigate how precisely d can be estimated and what determines this precision. We simulated data with different group sizes and estimated d using a mixed model that included IGE and d. Schemes included various average group sizes (4, 6, and 8), variation in group size (coefficient of variation (CV) ranging from 0.125 to 1.010), and three values of d (0, 0.5, and 1). A design in which individuals were randomly allocated to groups was used for all schemes and a design with two families per group was used for some schemes. Parameters were estimated using restricted maximum likelihood (REML). Bias and precision of estimates were used to assess their statistical quality. The dilution parameter of IGE can be estimated for simulated data with variation in group size. For all schemes, the length of confidence intervals ranged from 0.114 to 0.927 for d, from 0.149 to 0.198 for variance of DGE, from 0.011 to 0.086 for variance of IGE, and from 0.310 to 0.557 for genetic correlation between DGE and IGE. To estimate d, schemes with groups composed of two families performed slightly better than schemes with randomly composed groups. Dilution of IGE was estimable, and in general its estimation was more precise when CV of group size was larger. All estimated parameters were unbiased. Estimation of dilution of IGE allows the contribution of direct and indirect variance components to heritable variance to be quantified in relation to group size and, thus, it could improve prediction of the expected response to selection in environments with group sizes that differ from the average size.


International illicit drug traffic: the United States response.
The Government of the United States of America recognizes the serious threat of illicit drug abuse and trafficking and the associated economic costs and social destruction. The United States Government is waging a major battle to fight drug abuse and is working to lower demand at home through education and prevention, detoxification and treatment, and research as well as to lower supply through bilateral and multilateral crop eradication programmes. The Government is also working to lower supply within the United States through a campaign of illicit drug traffic interdiction of unprecedented size and scope. Among the actions taken to counter the illegal trade were: the creation of the South Florida Task Force headed by the Vice President; the establishment of 12 Organized Crime/Drug Enforcement task forces around the country; the setting up of the National Narcotics Border Interdiction System; the increased use of military resources to assist law enforcement agencies; and the initiation of a widespread cannabis eradication programme involving aerial spraying of paraquat.


Urea loading enhances freezing survival and postfreeze recovery in a terrestrially hibernating frog.
We tested the hypothesis that urea, an osmolyte accumulated early in hibernation, functions as a cryoprotectant in the freeze-tolerant wood frog, Rana sylvatica. Relative to saline-treated, normouremic (10 micromol ml(-1)) frogs, individuals rendered hyperuremic (70 micromol ml(-1)) by administration of an aqueous urea solution exhibited significantly higher survival (100% versus 64%) following freezing at -4 degrees C, a potentially lethal temperature. Hyperuremic frogs also had lower plasma levels of intracellular proteins (lactate dehydrogenase, creatine kinase, hemoglobin), which presumably escaped from damaged cells, and more quickly recovered neurobehavioral functions following thawing. Experimental freezing-thawing did not alter tissue urea concentrations, but did elevate glucose levels in the blood and organs of all frogs. When measured 24 h after thawing commenced, glucose concentrations were markedly higher in urea-loaded frogs as compared to saline-treated ones, possibly because elevated urea retarded glucose clearance. Like other low-molecular-mass cryoprotectants, urea colligatively reduces both the amount of ice forming within the body and the osmotic dehydration of cells. In addition, by virtue of certain non-colligative properties, it may bestow additional protection from freeze-thaw damage not afforded by glucose.


Efficient downregulation of ErbB-2 induces TACC1 upregulation in breast cancer cell lines.
The ErbB-2 gene, whose overexpression is observed in many types of tumors including breast cancer, plays an important role in carcinoma formation. Dysregulation of the human transforming acidic coiled-coil 1 (TACC1) and ErbB-2 genes is thought to be important in the development and progression of breast cancer. However, a putative interaction between ErbB-2 and TACC1 remains undetermined in breast cancer. After infecting BT474 cells with lentiviral-mediated ErbB2-specific shRNA, we detected the expression of ErbB-2 and TACC1 by real-time PCR and western blotting. ErbB-2 mRNA expression was decreased in the Lenti-ShERBB2 infected cells, and western blotting indicated a concordant reduction in ErbB-2 protein. TACC1 expression at the mRNA and protein levels was significantly upregulated by ErbB-2 silencing in BT474 cells. CCK-8 assay indicated that the inhibition of ErbB-2 expression increased the sensitivity of BT474 cells to docetaxel treatment. These findings provide proof and the foundation for the molecular and biological relationships of ErbB-2 and TACC1 in breast cancer.


[The activity of gas metabolism, thermoregulation, and antioxidant enzymes in aging C57Bl/6 mice].
The distribution type and correlative links between physiological and biochemical indices characterizing functional condition of the systems of gaseous exchange (V(O2) and V(CO2)), thermoregulation (body temperature and coefficient of thermoconductivity) and antioxidant defense have been studied in 62 young (3-5 mo.) and 58 old (23-26 mo.) male C57Bl/ 6 mice. The coefficients of variation differed significantly depending on the variable but not the age-group. Mean values of V(O2) and V(CO2), body temperature and thermoconductivity, but not activities of the antioxidant enzymes, declined in aging. Moreover, the activities of catalase, glutathione-peroxidase and glutathionereductase, i.e. enzymes involved in regulation of hydrogen peroxide level, increased in aging. The correlations between V(O2) and V(CO2), V(O2) and body temperature or V(O2) and the liver pH, as well as between the antioxidant enzyme activities exhibited little age-changes. However, three-dimensional non-linear models revealed significant age-changes in relations between the studied variables.


[The Mediterranean diet and cardiovascular disease: results of a pilot study from the SUN project (University of Navarre Follow-Up Study)].
The Mediterranean diet has been postulated as a model for the prevention of coronary heart disease. The SUN project ("Seguimiento Universidad de Navarra") is an epidemiological prospective cohort study of University of Navarre alumni started in 2000 with the main objective of identifying dietary and non-dietary determinants of these disorders. The feasibility of the project was verified in a two-phase pilot study. Volunteers participated in the first phase of the pilot study. Dietary and non-dietary exposures were measured using self-administered questionnaires and interview. A random sample of 600 graduates was selected for the second phase of the pilot study. The selected alumni received a questionnaire by mail. The 10th, 25th, 50th, 75th and 90th percentiles were calculated for the consumption of each food item or nutrient to estimate the inter-subject variability. The participants understood and answered the questionnaires properly. The mean time spent in completing the questionnaire was 55 minutes (95% CI: 50.7-59.3). A wide inter-subject variability was found in the consumption of food items considered as more representative of the Mediterranean diet (olive oil, fruits, vegetables and wine) with major differences between the 10th and 90th percentile. A Mediterranean diet pattern was found with a wide inter-subject variability. The response rate was acceptable. This finding is likely to ensure the identification of associations between the consumption of given food items and the occurrence of the main outcomes we are targeting.


Automated search of natively folded protein fragments for high-throughput structure determination in structural genomics.
Structural genomic projects envision almost routine protein structure determinations, which are currently imaginable only for small proteins with molecular weights below 25,000 Da. For larger proteins, structural insight can be obtained by breaking them into small segments of amino acid sequences that can fold into native structures, even when isolated from the rest of the protein. Such segments are autonomously folding units (AFU) and have sizes suitable for fast structural analyses. Here, we propose to expand an intuitive procedure often employed for identifying biologically important domains to an automatic method for detecting putative folded protein fragments. The procedure is based on the recognition that large proteins can be regarded as a combination of independent domains conserved among diverse organisms. We thus have developed a program that reorganizes the output of BLAST searches and detects regions with a large number of similar sequences. To automate the detection process, it is reduced to a simple geometrical problem of recognizing rectangular shaped elevations in a graph that plots the number of similar sequences at each residue of a query sequence. We used our program to quantitatively corroborate the premise that segments with conserved sequences correspond to domains that fold into native structures. We applied our program to a test data set composed of 99 amino acid sequences containing 150 segments with structures listed in the Protein Data Bank, and thus known to fold into native structures. Overall, the fragments identified by our program have an almost 50% probability of forming a native structure, and comparable results are observed with sequences containing domain linkers classified in SCOP. Furthermore, we verified that our program identifies AFU in libraries from various organisms, and we found a significant number of AFU candidates for structural analysis, covering an estimated 5 to 20% of the genomic databases. Altogether, these results argue that methods based on sequence similarity can be useful for dissecting large proteins into small autonomously folding domains, and such methods may provide an efficient support to structural genomics projects.


Differential nuclease sensitivity of the ovalbumin and beta-globin chromatin regions in erythrocytes and oviduct cells of laying hen.
We have monitored the differential nuclease sensitivity of defined regions of the chicken genome in different cells using a method which combines restriction enzyme digestion and blotting to diazobenzyloxymethyl (DBM)-paper (see Ref. 11). By using different specific probes and by scanning the bands on the autoradiograms, it is possible to compare on the same blot the digestion patterns of similar-sized fragments from different regions of the genome corresponding to "active" and reference "inactive" genes. We have demonstrated the preferential sensitivity to DNaseI and micrococcal nuclease digestion of the ovalbumin gene region in hen oviduct chromatin. The beta-globin gene region (containing both an adult and an embryonic gene) is also preferentially digested by DNaseI in hen mature erythrocyte nuclei, but at a lower rate than the ovalbumin gene region in oviduct. These observations raise the possibility that there may be several types of preferential nuclease sensitivities, all characterized by increased rates of digestion but to different levels, the highest corresponding to the very actively transcribing genes.