Serological testing for herpes simplex virus (HSV)-1 and HSV-2 infection.
Serological tests for herpes simplex virus (HSV) that can accurately distinguish between HSV-1 and HSV-2 are now commercially available. These tests detect antibodies to HSV glycoproteins G-1 and G-2, which evoke a type-specific antibody response. Focus Technologies produces the HerpeSelect-1 and HerpeSelect-2 enzyme-linked immunosorbent assay tests and the HSV-1 and HSV-2 HerpeSelect1/2 Immunoblot. Diagnology has marketed POCkit-HSV-2, a point-of-care test for HSV-2 that allows blood from a finger stick to be tested in a clinic. These tests can be used to confirm a genital herpes diagnosis, establish diagnosis of HSV infection in patients with atypical complaints, identify asymptomatic carriers, and identify persons at risk for acquiring HSV. Potential settings for use of these tests include sexually transmitted disease clinics, prenatal clinics, and clinics that care for patients with human immunodeficiency virus. Patient interest in HSV serological tests appears high.


Eptastigmine, nicotinamide and nicotinic acid determination using an inhibition enzyme sensor; application to pharmaceutical analysis.
An enzyme inhibition biosensor, developed in our laboratory and previously used for the analysis of compounds with anticholinesterase activity (e.g. physostigmine, neostigmine, pyridostigmine nicotine and organophosphorus compounds) has now been tested for the analysis of another recently synthesized cholinesterase inhibitor, i.e. eptastigmine. In addition nicotinic acid and nicotinamide, although displaying weaker inhibition properties, were also tested in pharmaceutical products using the same inhibition enzyme sensor. The biosensor consisted of a hydrogen peroxide amperometric electrode coupled to a functionalised nylon membrane chemically bonding both the enzymes butyrylcholinesterase and choline oxidase; a butyrylcholine standard solution in glycine buffer acted as substrate. The response of the system to all the inhibitors considered was characterised completely and the analysis of several pharmaceutical formulations containing nicotinamide or nicotinic acid was also performed.


Multisystem Inflammatory Syndrome in an Adult Who Received Casirivimab-Imdevimab (REGEN-COV).
Multisystem inflammatory syndrome in adults (MIS-A) is a systemic inflammatory syndrome that presents with a heterogeneous collection of signs and symptoms in adults. Here we present a case of a 38-year-old male who met the case definition of the MIS-A four weeks after a mild, symptomatic case of coronavirus disease 2019 (COVID-19) despite receiving casirivimab-imdevimab (REGEN-COV). Given the presence of signs and symptoms consistent with MIS-A, the patient was started on intravenous immune globulin (IVIG) and IV methylprednisolone. He promptly demonstrated clinical improvement over the next several days.


[Problems of ante mortem diagnostics of prion diseases].
The review presents the state-of-the-art on the problem of diagnosis of prion diseases (PD) in humans and animals with a brief description of their etiology and pathogenesis. We pointed out that understanding the nature of the etio logical agent of PD determined their zoonotic potential and led to the development of highly specific immunological diagnostic methods aimed at identifying the infectious isoform of prion protein (PrPd) as the only marker of the disease. In this regard, we briefly summarize the results of studies, including our own, concerning the conversion of normal prion protein molecules (PrPc) to PrPd, the production of monoclonal antibodies and their application as immunodiagnostic reagents for the post-mortem detection of PrPd in various formats of immunoassay. We also emphasize the issues related to the development of methods for ante mortem diagnostics of PD. In this regard, a method for amplifying amino acid sequences using quacking-induced conversion of PrPc to PrPd in real time (RTQuIC) described in details. The results of recent studies on the assessment of the sensitivity, specificity and reproducibility of this method, carried out in various laboratories around the world, are presented. The data obtained indicate that RT-QuIC is currently the most promising laboratory assay for detecting PrPd in biological material at the preclinical stage of the disease. The significant contribution of US scientists to the introduction of this method into clinical practice on the model of diagnosis of chronic wasting disease of wild Cervidae (CWD) is noted. The possible further spread of CWD in the population of moose and deer in the territories bordering with Russia, as well as the established fact of alimentary transmission of CWD to macaques, indicate the threat of the appearance of PD in our country. In conclusion, the importance of developing new hypersensitive and/or selective components of known methods for PrPd identification from the point of view of assessing the risks of creating artificial infectious prion proteins in vivo or in vitro, primarily new pathogenic isoforms ("strains") and synthetic prions, was outlined.


[Effect of beta-carotene on 60Co-gamma-induced mutation at T-lymphocyte hypoxanthine-guanine phosphoribosyl transferase locus in rats].
To study the effect of beta-carotene (beta-Car) on radiation-induced mutation. Mutant frequency at T-lymphocyte hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in rats was measured using a T-lymphocyte cloning technique and malondialdehyde (MDA) was measured using a thiobarbituric acid (TBA). Mutant frequency at the HGPRT locus was elevated by 60Co-gamma 3.75 Gy, and was reduced by ig beta-car 20 and 10 mg.kg-1 (P < 0.05), but not by beta-Car 5 mg.kg-1. It showed a dose-dependent relationship. MDA was signifcantly reduced in plasma of rats given beta-Car after 60 Co-gamma radiation. There was positive coherence between MDA and mutant frequency at the HGPRT locus (r = 0.9978, P < 0.05). Radiation-induced mutation is inhibited by beta-Car, associated with antioxidant action of beta-Car.


Parathyroid hormone alleviates non-alcoholic liver steatosis via activating the hepatic cAMP/PKA/CREB pathway.
Non-alcoholic fatty liver disease (NAFLD), hallmarked by liver steatosis, is becoming a global concern, but effective and safe drugs for NAFLD are still lacking at present. Parathyroid hormone (PTH), the only FDA-approved anabolic treatment for osteoporosis, is important in calcium-phosphate homeostasis. However, little is known about its potential therapeutic effects on other diseases. Here, we report that intermittent administration of PTH ameliorated non-alcoholic liver steatosis in diet-induced obese (DIO) mice and db/db mice, as well as fasting-induced hepatic steatosis. In vitro, PTH inhibits palmitic acid-induced intracellular lipid accumulation in a parathyroid hormone 1 receptor (PTH1R)-dependent manner. Mechanistically, PTH upregulates the expression of genes involved in lipid β-oxidation and suppresses the expression of genes related to lipid uptake and de novo lipogenesis by activating the cAMP/PKA/CREB pathway. Taken together, our current finding proposes a new therapeutic role of PTH on NAFLD.


Best's vitelliform macular dystrophy in 10- and 31-month-old siblings.
There are limited reports of Best's disease in children younger than 3 years. This is a case series of two family members aged 10 and 31 months with Best's disease. To the authors' knowledge, the 10-month-old male infant is the youngest patient with Best's disease documented by fundus photography.


Molecular mechanisms of primary resistance to flucytosine in Candida albicans.
Primary resistance in Candida albicans to flucytosine (5-FC) was investigated in 25 strains by identifying and sequencing the genes FCA1, FUR1, FCY21, and FCY22, which code for cytosine deaminase, uracil phosphoribosyltransferase (UPRT), and two purine-cytosine permeases, respectively. These proteins are involved in pyrimidine salvage and 5-FC metabolism. An association between a polymorphic nucleotide and resistance to 5-FC was found within FUR1 where the substitution of cytidylate for thymidylate at nucleotide position 301 results in the replacement of arginine with cysteine at amino acid position 101 in UPRT. Isolates that are homozygous for this mutation display increased levels of resistance to 5-FC, whereas heterozygous isolates have reduced susceptibility. Three-dimensional protein modeling of UPRT suggests that the Arg101Cys mutation disturbs the quaternary structure of the enzyme, which is postulated to compromise optimal enzyme activity. A single resistant isolate, lacking the above polymorphism in FUR1, has a homozygous polymorphism in FCA1 that results in a glycine-to-aspartate substitution at position 28 in cytosine deaminase.


Comparative transcriptome analysis of the human endocervix and ectocervix during the proliferative and secretory phases of the menstrual cycle.
Despite extensive studies suggesting increased susceptibility to HIV during the secretory phase of the menstrual cycle, the molecular mechanisms involved remain unclear. Our goal was to analyze transcriptomes of the endocervix and ectocervix during the proliferative and secretory phases using RNA sequencing to explore potential molecular signatures of susceptibility to HIV. We identified 202 differentially expressed genes (DEGs) between the proliferative and secretory phases of the cycle in the endocervix (adjusted p < 0.05). The biofunctions and pathways analysis of DEGs revealed that cellular assembly and epithelial barrier function in the proliferative phase and inflammatory response/cellular movement in the secretory phase were among the top biofunctions and pathways. The gene set enrichment analysis of ranked DEGs (score = log fold change/p value) in the endocervix and ectocervix revealed that (i) unstimulated/not activated immune cells gene sets positively correlated with the proliferative phase and negatively correlated with the secretory phase in both tissues, (ii) IFNγ and IFNα response gene sets positively correlated with the proliferative phase in the ectocervix, (iii) HIV restrictive Wnt/β-catenin signaling pathway negatively correlated with the secretory phase in the endocervix. Our data show menstrual cycle phase-associated changes in both endocervix and ectocervix, which may modulate susceptibility to HIV.


Study of cellular changes induced by moderate cerebral ischemia achieved through internal carotid artery ligation.
Reduced cerebral blood flow beyond the compensatory mechanisms leads to cerebral hypoxia. Hypoxia causes various lesions of neurons, glial cells and cerebral blood vessels, depending on its duration and intensity. In our study, we reduced cerebral blood flow in the experience animal on average by 30%, by right internal carotid artery ligation. Fifteen days after the onset of hypoxia, by histology and immunohistochemical studies, we identified neuronal, glial and vascular damage. Lesions of nerve and glial cells ranged from changes of cytoplasmic tinting with the development of "red neurons", to neuronal and glial cytolysis with areas of focal necrosis. Vascular lesions were represented by the collapse, fragmentation and discontinuity of capillaries, always associated with a marked perivascular edema.


An Augmented Reality magic mirror as additive teaching device for gross anatomy.
When preparing young medical students for clinical activity, it is indispensable to acquaint them with anatomical section images which enable them to use the clinical application of imaging methods. A new Augmented Reality Magic Mirror (AR MM) system, which provides the advantage of a novel, interactive learning tool in addition to a regular dissection course, was therefore tested and evaluated by 880 first-year medical students as part of the macroscopic anatomy course in 2015/16 at Ludwig-Maximilians-Universität (LMU) in Munich. The system consists of an RGB-D sensor as a real-time tracking device, which enables the system to link a deposited section image to the projection of the user's body, as well as a large display mimicking a real-world physical mirror. Using gesture input, the users have the ability to interactively explore radiological images in different anatomical intersection planes. We designed a tutorial during which students worked with the system in groups of about 12 and evaluated the results. Subsequently, each participant was asked to assess the system's value by filling out a Likert-scale questionnaire. The respondents approved all statements which stressed the potential of the system to serve as an additional learning resource for anatomical education. In this case, emphasis was put on active learning, 3-dimensional understanding, and a better comprehension of the course of structures. We are convinced that such an AR MM system can be beneficially installed into anatomical education in order to prepare medical students more effectively for the clinical standards and for more interactive, student-centered learning.


Atamestane, a new aromatase inhibitor for the management of benign prostatic hyperplasia.
Atamestane is a new, competitive, and irreversible inhibitor of estrogen biosynthesis. Its pharmacologic action has been evaluated in mice, rats, rabbits, dogs, monkeys, and humans. In rats, atamestane leads to a decrease of pregnant mare serum gonadotropin-stimulated ovarian estrogen production, and inhibits androstenedione-induced estrogenic effects such as uterine growth and abortion. In all species tested, atamestane lacks other intrinsic hormonal or antihormonal activities, and shows no inhibition of other cytochrome P450-dependent enzymes of adrenal steroidogenesis. However, it inhibits estrogen-related negative feedback. The extent and consequences of the induced counterregulation of the pituitary-hypothalamic axis show major sex- and species-specific differences. Atamestane is highly effective in inhibiting estrogen-induced hyperplastic changes in the fibromuscular stroma of the prostate in androstenedione-treated dogs and monkeys. In male volunteers and patients with benign prostatic hyperplasia (BPH), atamestane induces an expected reduction of serum (and BPH tissue) estrogen concentrations without significant changes in androgen levels. In conclusion, all available results indicate that atamestane is a selective (no inhibition of adrenal function), pure (no endocrine side effects), and highly effective steroidal aromatase inhibitor, with an excellent safety profile. Based on the discussion of its clinical potential, atamestane seems to be a promising compound for the management of BPH.


Diffuse Alveolar Hemorrhage Synergistically Induced by Amiodarone and Rheumatoid Arthritis.
Diffuse alveolar hemorrhage (DAH) syndrome is characterized by bleeding into the alveolar spaces of the lungs and occurs when there is an injury to the alveolar microcirculation that leads to hemorrhage from the alveolar capillaries. We report a case of an 82-year-old woman who presented with acute respiratory distress. The patient had a history of rheumatoid arthritis (RA) and was on amiodarone for atrial fibrillation therapy. Initial diagnostic workup, including bronchoscopy and imaging studies, revealed diffuse opacities and the bronchoalveolar lavage fluid consistent with DAH. The patient required aggressive management with supportive care and corticosteroids. Laboratory work showed a synergistic effect between amiodarone and RA in inducing DAH. This makes the report unique as no reports in the literature described a synergic effect of amiodarone and RA in inducing DAH. The primary objective of this report is to guide physicians and remind them to keep DAH at the top of their differential diagnosis in the setting of an RA patient taking amiodarone.


From PIM1 to PI3Kδ via GSK3β: Target Hopping through the Kinome.
Selective inhibitors of phosphoinositide 3-kinase delta are of interest for the treatment of inflammatory diseases. Initial optimization of a 3-substituted indazole hit compound targeting the kinase PIM1 focused on improving selectivity over GSK3β through consideration of differences in the ATP binding pockets. Continued kinase cross-screening showed PI3Kδ activity in a series of 4,6-disubstituted indazole compounds, and subsequent structure-activity relationship exploration led to the discovery of an indole-containing lead compound as a potent PI3Kδ inhibitor with selectivity over the other PI3K isoforms.


Aminoribosylated Analogues of Muraymycin Nucleoside Antibiotics.
Nucleoside antibiotics are uridine-derived natural products that inhibit the bacterial membrane protein MraY. MraY is a key enzyme in the membrane-associated intracellular stages of peptidoglycan biosynthesis and therefore considered to be a promising, yet unexploited target for novel antibacterial agents. Muraymycins are one subclass of such naturally occurring MraY inhibitors. As part of structure-activity relationship (SAR) studies on muraymycins and their analogues, we now report on novel derivatives with different attachment of one characteristic structural motif, i.e., the aminoribose moiety normally linked to the muraymycin glycyluridine core unit. Based on considerations derived from an X-ray co-crystal structure, we designed and synthesised muraymycin analogues having the aminoribose attached (via a linker) to either the glycyluridine amino group or to the uracil nucleobase. Reference compounds bearing the non-aminoribosylated linker units were also prepared. It was found that the novel aminoribosylated analogues were inactive as MraY inhibitors in vitro, but that the glycyluridine-modified reference compound retained most of the inhibitory potency relative to the unmodified parent muraymycin analogue. These results point to 6'-N-alkylated muraymycin analogues as a potential novel variation of the muraymycin scaffold for future SAR optimisation.


Regulation of T cell responses in the developing human fetus.
Although human T cells enter the peripheral lymphoid tissues early during fetal development, the adaptive immune system in the fetus has largely been regarded as functionally immature and unresponsive to stimulation. In this study, we show that depletion of fetal CD4+CD25(high) T regulatory (T(Reg)) cells, which are present at high frequency in fetal lymphoid tissues, results in vigorous T cell proliferation and cytokine production in vitro, even in the absence of exogenous stimulation. Analysis of CD4+ and CD8(+) T cell populations revealed a large subset of cells that expressed the early activation Ag, CD69. We show that this population represents a subset of highly reactive fetal T cells actively suppressed by fetal CD4+CD25(high) T(Reg) cells during development. These findings indicate that fetal T cells are, in the absence of CD4+CD25(high) T(Reg) cells, highly responsive to stimulation and provide evidence for an important role for CD4+CD25(high) T(Reg) cells in controlling T cell responses in utero.


Comparison of Facial Muscle Activation Patterns Between Healthy and Bell's Palsy Subjects Using High-Density Surface Electromyography.
Facial muscle activities are essential for the appearance and communication of human beings. Therefore, exploring the activation patterns of facial muscles can help understand facial neuromuscular disorders such as Bell's palsy. Given the irregular shape of the facial muscles as well as their different locations, it should be difficult to detect the activities of whole facial muscles with a few electrodes. In this study, a high-density surface electromyogram (HD sEMG) system with 90 electrodes was used to record EMG signals of facial muscles in both healthy and Bell's palsy subjects when they did different facial movements. The electrodes were arranged in rectangular arrays covering the forehead and cheek regions of the face. The muscle activation patterns were shown on maps, which were constructed from the Root Mean Square (RMS) values of all the 90-channel EMG recordings. The experimental results showed that the activation patterns of facial muscles were distinct during doing different facial movements and the activated muscle regions could be clearly observed. Moreover, two features of the activation patterns, 2D correlation coefficient (corr2) and Centre of Gravity (CG) were extracted to quantify the spatial symmetry and the location of activated muscle regions respectively. Furthermore, the deviation of activated muscle regions on the paralyzed side of a face compared to the healthy side was quantified by calculating the distance between two sides of CGs. The results revealed that corr2 of the activated facial muscle region (classified into forehead region and cheek region) in Bell's palsy subjects was significantly (p < 0.05) lower than that in healthy subjects, while CG distance of activated facial region in Bell's palsy subjects was significantly (p < 0.05) higher than that in healthy subjects. The correlation between corr2 of these regions and Bell's palsy [assessed by the Facial Nerve Grading Scale (FNGS) 2.0] was also significant (p < 0.05) in Bell's palsy subjects. The spatial information on activated muscle regions may be useful in the diagnosis and treatment of Bell's palsy in the future.


Smoking and Cancers: Case-robust Analysis of a Classic Data Set.
A typical structural equation model is intended to reproduce the means, variances, and correlations or covariances among a set of variables based on parameter estimates of a highly restricted model. It is not widely appreciated that the sample statistics being modeled can be quite sensitive to outliers and influential observations leading to bias in model parameter estimates. A classic public epidemiological data set on the relation between cigarette purchases and rates of four types of cancer among states in the USA is studied with case-weighting methods that reduce the influence of a few cases on the overall results. The results support and extend the original conclusions; the standardized effect of smoking on a factor underlying deaths from bladder and lung cancer is .79.


Emotional experience and facial expression in Alzheimer's disease.
Emotional experiences and facial muscle activity of individuals with Alzheimer's Disease (AD) (n = 13) and healthy elderly control subjects (n = 21) were measured while viewing emotion-eliciting images. Alzheimer's disease and control groups rated their emotional experiences similarly and in the expected directions on dimensions of valence and arousal. Change in corrugator activity while viewing images, compared to baseline, was comparable across groups and was greatest while viewing negative images. Change in zygomatic activity, however, was significantly different between AD and control groups, with AD subjects demonstrating an inverted pattern of activity compared to controls. These findings are discussed as possible consequences of frontal cortical system involvement accompanying the disease process.


[Characteristics of HIV's sexual behavior and their effect on the secondary transmission rate].
To analyze the sexual behavior characteristics of the sexually transmitted HIV-positive people and to estimate the secondary transmission rate. Field investigation and literature review were conducted among sexually transmitted HIV-positive people to collect general information during the last 6 months before notification of HIV infection. A mathematical model was used to estimate the secondary transmission rate. A total of 769 HIV-positive people were recruited for the study, 186 of them were women, 286 were men who have sex with women (MSW), 242 were men who have sex with men (MSM) and 55 were men who have sex with men and women (MSMW). During 6 months, the average sex partner among these 4 groups were 2.29, 1.61, 3.32 and 4.10, respectively; the sexual behavior frequency were 26.03, 20.97, 14.77 and 25.51, respectively; the rates of non-use of condom were 74.14%, 73.53%, 59.60% and 72.06%, respectively; the secondary transmission rate were 0.0095, 0.0151, 0.1759 and 0.1985, respectively. Under constant conditions of other factors, the secondary transmission rates decreased by 30.13%-82.00%, 23.00%-49.51%, and 16.10%-19.09%, respectively, if there was a reduction in 1 sex partner, 1 time/month for the sexual frequency and 20% of rate for non-use of condom. The HIV secondary transmission from MSMW HIV-positive people to general population was the highest. Change in sexual behavior of the HIV-positive people can decrease the HIV secondary transmission rate significantly.


Comparison between Sonographic Features and Fine Needle Aspiration Cytology with Histopathology in the Diagnosis of Solitary Thyroid Nodule.
High resolution ultrasonography (USG) is the first-line investigation in evaluation of euthyroid nodules. Thyroid imaging reporting and data system (TIRADS) is an USG-based risk stratification system for classifying thyroid nodules. Subjects with high-risk category of TIRADS undergo fine needle aspiration cytology (FNAC) and FNAC findings are reported according to Bethesda classification. Bethesda categories are used for determining risk of malignancy. Data regarding sonographic classification of thyroid nodule and its cytological association with respect to final histopathological diagnosis remains scarcely available in India. The study evaluated euthyroid nodules for risk of malignancy and compared sonographic features and FNAC (Bethesda classification) findings with histopathology of excised samples. This was a single-center observational study on 137 consecutive subjects of solitary euthyroid nodule. All subjects underwent USG according to TIRADS and FNAC where applicable. Surgical biopsy report was used as a gold standard. The sensitivity, specificity, accuracy, positive predictive and negative predictive value of FNAC were 80%, 90%, 85%, 86%, and 86.6% and TIRADS were 80%, 47.2% 61%, 51.3%, and 77.3%, respectively. FNAC classification was equally sensitive and more specific than TIRADS. Among individual USG parameters, micro-calcification was most sensitive (80%) and specific (86%). Irregular margin and taller-than-wider shape had a specificity of 89% and 92%, respectively. 3 patients (14.28%) with benign cytology and suspicious USG features (specifically TIRADS 4 & 5) undergoing surgery had malignancy in final HPE. USG and FNAC are equally sensitive in diagnosing malignant thyroid nodule but FNA is more specific (90%). It's a minimally invasive method which can be used to distinguish malignant from benign lesions with a high degree of accuracy (85%). In patient having high risk feature on USG, a benign cytology needs to be repeat FNAC and they should undergo surgical biopsy for confirmation.


In vivo inhibition of granulopoiesis in acute inflammation requires T lymphocyte integrity.
In previous studies, we have shown that mice undergoing an inflammatory reaction induced by subcutaneous (s.c.) implantation of copper rods elaborate humoral factors that initially enhance, and subsequently inhibit, diffusion chamber (DC) granulopoiesis. In order to quantify the inhibition of DC granulopoiesis after inflammation, one to three copper rods were implanted s.c. either at the same place (1 abscess), or at different sites (multiple abscesses). There was an inverse relationship between the increase in the number of abscesses, and the number of DC granulocytic cells measured in the inhibitory phase. To investigate the role of T lymphocytes in the release of putative inhibitory factor(s) that act on DC cells, cyclosporin A (CyA), a T lymphocyte function inhibitor, was given orally each day (0.75 mg) to mice, starting two days before copper implantation. CyA abrogated the inflammation-related inhibition on DC spleen colony-forming units (CFU-s), granulocyte-macrophage colony-forming units (CFU-gm), and total cell production. To confirm these results, DC were implanted in T cell deficient nude mice where no inhibition of DC cells was observed after inflammation. In conclusion, our data suggest that the in vivo inhibition of granulopoiesis is related to the level of the inflammatory stimulus and requires the functional integrity of T cells.


Risk factors for multisymptom illness in US Army veterans of the Gulf War.
This research study examined the prevalence of symptoms and identified risk factors for reported symptoms among a group of Army Gulf War (GW) veterans. A survey was mailed to all members of the Ft. Devens cohort in 1997, representing the third assessment of a group that consisted of 2949 US Army soldiers deployed to the Gulf, and was studied initially in 1991. A total of 1290 subjects responded to the mailed survey; aggressive follow-up methods to address non-response bias were employed. Subjects were classified as having multisymptom illness if they reported symptoms from at least two of three symptom categories (fatigue, mood-cognition, musculoskeletal). Sixty percent of the respondents met criteria for multisymptom illness. Female gender, lower levels of education, psychological symptoms, self-reported use of a medical clinic in the Gulf, ingestion of anti-nerve gas pills (pyridostigmine bromide), anthrax vaccination, tent heaters, exposure to oil fire smoke, and chemical odors were significantly related to multisymptom illness in logistic regression analyses. Analyses in which subjects were stratified by level of psychological symptoms revealed different sets of GW-service environmental exposures and suggest that subgroups of GW veterans may have different sets of risk factors.


Use of bisphosphonates in a case of Perthes disease.
Bisphosphonates are a group of drugs that have been used extensively for more than 10 years to treat adults with osteoporosis. Use of bisphosphonates in pediatrics has generally been confined to conditions such as osteogenesis imperfecta or other conditions with bone density problems. Bisphosphonates increase bone density, bone mineral content, and strength; improve mobility; reduce fracture rates; and are effective agents in combating bone pain. Bisphosphonates inhibit osteoclastic action, thereby increasing bone density (Rauch & Glorieux 2004). Recently there has been interest in the effect of bisphosphonates in localized disorders of bone, including avascular necrosis. The third-generation bisphosphonate zoledonic acid is currently being explored as a treatment for children presenting with Perthes disease at the Children's Hospital at Westmead, Sydney, Australia, and this is a report of that experience.


Greater Social Competence Is Associated With Higher Interpersonal Neural Synchrony in Adolescents With Autism.
Difficulty engaging in reciprocal social interactions is a core characteristic of autism spectrum disorder. The mechanisms supporting effective dynamic real-time social exchanges are not yet well understood. This proof-of-concept hyperscanning electroencephalography study examined neural synchrony as the mechanism supporting interpersonal social interaction in 34 adolescents with autism spectrum disorder (50% female), age 10-16 years, paired with neurotypical confederates of similar age. The degree of brain-to-brain neural synchrony was quantified at temporo-parietal scalp locations as the circular correlation of oscillatory amplitudes in theta, alpha, and beta frequency bands while the participants engaged in a friendly conversation. In line with the hypotheses, interpersonal neural synchrony was significantly greater during the social interaction compared to the baseline. Lower levels of synchrony were associated with increased behavioral symptoms of social difficulties. With regard to sex differences, we found evidence for stronger interpersonal neural synchrony during conversation than baseline in females with autism, but not in male participants, for whom such condition differences did not reach statistical significance. This study established the feasibility of hyperscanning during real-time social interactions as an informative approach to examine social competence in autism, demonstrated that neural coordination of activity between the interacting brains may contribute to social behavior, and offered new insights into sex-related variability in social functioning in individuals with autism spectrum disorders.


Nonsense Suppression Therapy: An Emerging Treatment for Hereditary Skin Diseases.
Nonsense mutations cause the premature termination of protein translation via premature termination codons (PTCs), leading to the synthesis of incomplete functional proteins and causing large numbers of genetic disorders. The emergence of nonsense suppression therapy is considered to be an effective method for the treatment of hereditary diseases, but its application in hereditary skin diseases is relatively limited. This review summarizes the current research status of nonsense suppression therapy for hereditary skin diseases, and discusses the potential opportunities and challenges of applying new technologies related to nonsense suppression therapy to dermatology. Further research is needed into the possible use of nonsense suppression therapy as a strategy for the safer and specific treatment of hereditary skin diseases.


Exploring biomolecular dynamics and interactions using advanced sampling methods.
Molecular dynamics (MD) and Monte Carlo (MC) simulations have emerged as a valuable tool to investigate statistical mechanics and kinetics of biomolecules and synthetic soft matter materials. However, major limitations for routine applications are due to the accuracy of the molecular mechanics force field and due to the maximum simulation time that can be achieved in current simulations studies. For improving the sampling a number of advanced sampling approaches have been designed in recent years. In particular, variants of the parallel tempering replica-exchange methodology are widely used in many simulation studies. Recent methodological advancements and a discussion of specific aims and advantages are given. This includes improved free energy simulation approaches and conformational search applications.


Role of PP2B in cAMP-induced dephosphorylation and translocation of NTCP.
cAMP-mediated stimulation of hepatic bile acid uptake is associated with dephosphorylation and translocation of Na+-taurocholate (TC) cotransporting peptide (NTCP) to the plasma membrane. Although translocation of NTCP may be facilitated by dephosphorylation, the mechanism of dephosphorylation is unknown. The ability of cAMP to translocate and dephosphorylate NTCP is, in part, dependent on cAMP-mediated increases in cytosolic Ca2+ concentration ([Ca2+]), indicating that a Ca2+/calmodulin-dependent protein phosphatase (PP2B) may be involved. Thus we studied the role of PP2B using the inhibitor cypermethrin (CM). Freshly isolated hepatocytes were pretreated with 1-5 nM CM for 30 min followed by 15 min incubation with 10 microM 8-(4-chlorophenylthio)cAMP. CM (5 nM) and FK-506 (5 microM) inhibited cAMP-stimulated TC uptake by 80 and 75%, respectively, without affecting basal TC uptake. CM also reversed cAMP-mediated NTCP dephosphorylation and translocation to 80 and 15% of the basal level, respectively. cAMP stimulated PP2B activity by 60%, and this effect was completely inhibited by 5 nM CM. PP2B dephosphorylated NTCP immunoprecipitated from control but not from cAMP-treated hepatocytes. The effect of CM was not due to any changes in cAMP-mediated increases in cytosolic [Ca2+] or decreases in mitogen-activated protein kinase (extracellular regulated kinases 1 and 2) activity. Taken together, these results suggest that cAMP dephosphorylates NTCP by activating PP2B in hepatocytes, and PP2B-mediated dephosphorylation of NTCP may be involved in cAMP-mediated NTCP translocation to the plasma membrane.


At the frontlines of digitisation: a qualitative study on the challenges and opportunities in maintaining accurate, complete and timely digital health records in India's government health system.
To understand factors underpinning the accuracy and timeliness of mobile phone numbers and other health information captured in India's government registry for pregnant and postpartum women. Accurate and timely registration of mobile phone numbers is necessary for beneficiaries to receive mobile health services. Madhya Pradesh and Rajasthan states in India at the community, clinical, and administrative levels of the health system. Interviews (n=59) with frontline health workers (FLHWs), data entry operators, and higher level officials. Focus group discussions (n=12) with pregnant women to discuss experiences with sharing data in the health system. Observations (n=9) of the process of digitization and of interactions between stakeholders for data collection. Thematic analysis identified how key actors experienced the data collection and digitisation process, reasons for late or inaccurate data, and mechanisms that can bolster timeliness and accuracy. Pregnant women were comfortable sharing mobile numbers with health workers, but many were unaware that their data moved beyond their FLHW. FLHWs valued knowing up-to-date beneficiary mobile numbers, but felt little incentive to ensure accuracy in the digital record system. Delays in registering pregnant women in the online portal were attributed to slow movement of paper records into the digital system and difficulties in gathering required documents from beneficiaries. Data, including women's phone numbers, were handwritten and copied multiple times by beneficiaries and health workers with variable literacy. Supervision tended to focus on completeness rather than accuracy. Health system actors noted challenges with the digital system but valued the broader project of digitisation. Increased focus on training, supportive supervision, and user-friendly data processes that prioritise accuracy and timeliness should be considered. These inputs can build on existing positive patient-provider relationships and health system actors' enthusiasm for digitisation.


Is Gaining Affective Commitment the Missing Strategy for Successful Change Management in Healthcare?
Despite the requirement for continual change and development, change failure is omnipresent in health care, ranging from small technical errors within new systems, processes or technologies, through to breakdowns and large-scale disaster. Despite decades of research investment, consultancy and initiatives, creating a healthcare context that promotes clinician engagement with change remains elusive, with limited demonstrated progress. Affective commitment to change refers to commitment that is driven by a desire to support change based on its perceived benefits or value, as opposed to commitment that is based on a sense of obligation or the minimization of costs. Recent evidence from health-care contexts indicates that affective commitment to change drives change readiness more so than the individual's self-efficacy for dealing with the change. Considering evidence regarding the effect of affective commitment to change on individual and collective change readiness among health-care staff, we may need to reorient our current strategies for managing change. We explore the opportunities to enhance affective commitment to change and, in turn, change readiness through adopting values-based approaches to designing and executing change proposals with clinicians and service users.


Plant oxygen sensing is mediated by the N-end rule pathway: a milestone in plant anaerobiosis.
Like all aerobic organisms, plants require molecular oxygen for respiratory energy production. In plants, hypoxic conditions can occur during natural events (e.g., flooding), during developmental processes (e.g., seed germination), and in cells of compact tissues with high metabolic rates. Plant acclimation responses to hypoxia involve a modulation of gene expression leading to various biochemical, physiological, and morphological changes that stave off eventual anoxia. In contrast with the animal kingdom, a direct oxygen-sensing mechanism in plants has been elusive so far. However, two recent independent studies show that oxygen sensing in plants operates via posttranslational regulation of key hypoxia response transcription factors by the N-end rule pathway. The N-end rule is an evolutionarily conserved pathway for protein degradation that relates the fate of a protein with the identity of its N-terminal residues. Results from these studies demonstrate that oxygen-dependent modification and targeted proteolysis of members of the ethylene response factor group VII transcription factor family regulate hypoxia-responsive gene expression in Arabidopsis thaliana. The discovery of this plant hypoxia-sensing mechanism sets the stage for further research on plant homeostatic response to oxygen, which could be relevant to understanding plant distributions in flood-prone ecosystems and improving hypoxia tolerance of crops.


Odontogenic myxoma in an 8-year-old girl: A case report with review of literature.
Aggressive enlargements of maxilla in pediatric patients are uncommon and present with diagnostic and therapeutic dilemma. The decision on therapeutic modality is based on an early and accurate diagnosis, minimizing disease-associated morbidity which is of utmost importance considering the young age and thereby resulting in better prognosis. Odontogenic myxoma is a locally aggressive lesion which is primarily seen in relation to odontogenic apparatus in mandibular posterior region in association with an impacted tooth. This presentation describes a unique case of odontogenic myxoma of anterior maxilla in an 8-year-old girl with emphasis on its diagnosis and treatment planning.


Understanding communication during hospitalist service changes: a mixed methods study.
Little data exist to inform hospitalist communication during service changes. To characterize hospitalist handoffs during service changes. Serial survey study. Single academic medical center. From May to December 2007, 60 service changes among 17 hospitalists on a nonteaching service were targeted for evaluation using an anonymous 18-item survey that was completed by hospitalists within 48 hours of assuming care for patients. Survey items assessed completeness of handoff communication, certainty of patient care plans, missed information, time spent recovering information, and near misses/adverse events due to incomplete handoffs. The association between completeness of communication and handoff outcomes was examined. Narrative comments were analyzed qualitatively. Ninety-three percent (56/60) of surveys were returned. All 17 hospitalists participated. Thirteen percent of respondents reported incomplete handoffs and 18% were uncertain of care plan on transition day. At least 1 near miss, attributable to incomplete communication was reported by 16%. Hospitalists who reported incomplete handoffs were more likely to report uncertainty about patient care plans on the transition day (71% incomplete vs. 10% complete, P < 0.01), discovery of missing information (71% incomplete vs. 24% complete, P = 0.01), near misses/adverse events (57% incomplete vs. 10% complete, P < 0.01), and more time resolving issues arising from missed information (71% incomplete vs. 22% complete, P < 0.01). Qualitative comments suggest the need for a more systematic, focused, team-based, and patient-centered handoff model. Incomplete handoffs during service changes are associated with uncertainty and potential patient harm. Suggestions to improve the completeness of hospitalist service change communications are offered.


Edge-setting criterion in confocal microscopy.
The intensity that locates the image of an edge in a confocal microscope is calculated as a function of detector size. Both pinhole and slit detectors are considered. Knowledge of this edge-setting criterion permits accurate determination of the edge position, as is necessary for confocal metrology.


Neuroactive Steroids in Acute Ischemic Stroke: Association with Cognitive, Functional, and Neurological Outcomes.
Despite several scientific and technological advances, there is no single neuroprotective treatment that can reverse the brain damage after acute ischemic stroke (AIS). Neuroactive steroids are cholesterol-derived hormones that have the ability to modulate the normal and pathologic nervous system employing genomic and nongenomic mechanisms. In this work, we first investigated if AIS affects the plasma concentration of 5 neuroactive steroids (cortisol, estradiol, progesterone, testosterone, and 3α-androstenediol glucuronide). Second, we studied if levels of circulating steroids associate with neurological, cognitive, and functional outcome in a cohort of 60- to 90 year-old male and female patients with AIS. For this purpose, we recruited patients who were hospitalized at the Emergency Room of the Central Military Hospital within the first 24 h after stroke onset. We designed 2 experimental groups, each one composed of 30 control subjects and 30 AIS patients, both males and females. The assessment of neurological deficit was performed with the NIHSS and the tests used for the functional and cognitive status were: (1) modified Rankin Scale; (2) Photo test, and (3) abbreviated Pfeiffer's mental status questionnaire. We observed a significant difference in plasma concentration of cortisol and estradiol between both experimental groups. In the AIS group, higher levels of these neuroactive steroids were associated with more pronounced neurological, cognitive and functional deficits in women compared to men. We propose that in elderly patients, high levels of circulating neuroactive steroids like cortisol and estradiol could potentiate AIS-mediated neuropathology in the ischemic and penumbra areas.


Time resolved mechanism of the isotope selectivity in the ultrafast light induced dissociation in N2.
The time evolution of a vacuum ultraviolet excited N2 molecule is followed all the way from an ultrafast excitation to dissociation by a quantum mechanical simulation. The primary aim is to discern the role of the excitation by a pulse short compared to the vibrational period, to discern the different coupling mechanisms between different electronic states, nonadiabatic, spin orbit, and to analyze the origin of any isotopic effect. We compare the picture in the time and energy domains. The initial ultrafast excitation pumps the molecule to a coherent electronic wave packet to which several singlet bound electronic states contribute. The total nonstationary wave function is given as a coherent sum of nuclear wave packets on each electronic state times the stationary electronic wave function. When the wave packets on different electronic states overlap, they are coupled in a mass-dependent manner whether one uses an adiabatic or a diabatic electronic basis. A weak spin-orbit coupling acts as a bottleneck between the bound singlet part of phase space and the triplet manifold of states in which dissociation takes place. To describe the spin-orbit perturbation that is ongoing in time, an energy-resolved eigenstate representation appears to be more intuitive. In the eigenstate basis, the singlet-to-triplet population transfer is large only between those vibronic eigenstates that are quasiresonant in energy. The states in resonance are different for different excitation energy ranges. The resonances are mass dependent, which explains the control of the isotope effect through the profile of the pulse.


Developmental changes in endogenous control of attention: the role of target familiarity on infants' distraction latency.
This study evaluated the interactive effects of endogenous and exogenous influences on infants' attention allocation by assessing the role of target familiarity on distraction latency during object exploration. In Experiment 1 (N = 54), infants' distraction latencies as they investigated both familiar toys (ones they previously had seen in a familiarization procedure) and novel toys (ones they had not seen in the familiarization procedure) were assessed longitudinally at 6.5 and 9 months of age. In Experiment 2 (N = 32), infants' distraction latencies were assessed at either 6.5 or 10 months as they investigated either familiar or novel targets. In both experiments, older infants, but not younger infants, exhibited longer latencies as they investigated novel toys as compared with their latencies as they investigated familiar toys. These results are discussed in terms of developmental changes in the interactive effects of endogenous and exogenous factors controlling attention allocation.


[Criticism of the concept of progress in encyclopedic psychiatry].
The concept of progress in biological psychiatry its being opposed in this study to another one which makes it necessary to adopt a different mode of thinking and to be ready for a change of paradigms. Both concepts are not exclusive of each other, yet, one cannot confer either of them an absolute value without limiting the subject and the methological tools necessary to psychiatric-psychopathological research. This will be explained, using everyday clinical cases.


Dependence of baroreceptor-mediated sympathetic outflow on biventricular assist device driving frequency.
To investigate the drive condition of the artificial heart from a neurophysiological point of view, the dynamic transduction characteristics of the baroreflex system were analyzed by means of sympathetic neurograms. Two pneumatically actuated ventricular assist devices were implanted as biventricular bypasses (BVBs) in adult mongrel dogs to compare the natural heart circulation and prosthetic circulation. After BVB pumping was initiated, the natural heart was electrically fibrillated. Renal sympathetic nerve activity (RSNA) was recorded to analyze sympathetic outflow. Coherence function between the arterial pressure and RSNA was calculated to evaluate the linearity of the baroreflex system. The largest peak coherency was observed when BVB was driven at the frequency of natural heart beat prior to electrical fibrillation, which suggests that the baroreflex system shows the largest linearity at this frequency. These results suggest the possibility that the natural heart beat frequency is the setting frequency at which the baroreflex system transfers the hemodynamic rhythm to the sympathetic outflow.


Interface shear microrheometer with an optically driven oscillating probe particle.
We report the first experimental demonstration of an active interfacial shear microrheometer (ISMR) that uses a particle trapped by oscillating optical tweezers (OT) to probe the shear modulus G(s)(*)(ω) of a gas/liquid interface. The most significant advantages of the oscillating OT in a rheology study are: (1) very high sensitivity compared to other active microrheology methods and (2) the ability to measure both the real and imaginary components of the complex shear modulus without relying on the use of Kramers-Kronig relation, which can be problematic at low frequencies for most of the passive methods. We demonstrate the utilities of our ISMR in two case studies: (1) a 1,2-dipalmitoyl-sn-glycero-3-phosphocholine monolayer and (2) a composite of poly(styrene sulfonate) and dioctadecyldimethylammonium at the air/water interface in regimes where no other active instruments can explore.


The cytokine and histological response in islet xenograft rejection is dependent upon species combination.
Islet xenografts have clinical potential, may avoid hyperacute rejection, and therefore are a good place to examine the cellular xenograft immune response. The aim of this study was to examine the cellular, humoral, and cytokine response in islet xenograft rejection and to determine the difference in the immune response with a different donor species. Two islet xenograft models (DA rat islets to B6AF1 mouse and canine islets to B6AF1 mouse) and a mouse syngeneic control model were examined histologically and by a semiquantitative polymerase chain reaction method. There was significant up-regulation of all intragraft cytokines tested (interleukin [IL]-2, IL-4, IL-5, IL-10, and interferon-gamma) in both xenograft models compared with the controls. However, the dog islet grafts had higher levels of IL-4 and IL-5 gene expression than the rat islet grafts, which, conversely, had higher levels of interferon-gamma gene expression. These differences correlated with the histological and anti-donor antibody production differences between the two models. The dog to mouse model had an intense eosinophilic infiltrate and an early up-regulation of anti-donor antibody, whereas there was little eosinophilic infiltrate and a delayed anti-donor antibody up-regulation in the rat to mouse model. The mouse used different mechanisms to reject the rat and canine islets, suggesting that the immune response in islet xenograft rejection may be dependent on the species combination. It may not be possible to characterize the cellular xenograft rejection response in a bipolar manner as has been the case with humoral rejection response. Caution therefore needs to be taken before extrapolating the cellular immune responses seen in animal models to the clinical setting.


No Associations between Interindividual Differences in Sleep Parameters and Episodic Memory Consolidation.
Sleep and memory are stable and heritable traits that strongly differ between individuals. Sleep benefits memory consolidation, and the amount of slow wave sleep, sleep spindles, and rapid eye movement sleep have been repeatedly identified as reliable predictors for the amount of declarative and/or emotional memories retrieved after a consolidation period filled with sleep. These studies typically encompass small sample sizes, increasing the probability of overestimating the real association strength. In a large sample we tested whether individual differences in sleep are predictive for individual differences in memory for emotional and neutral pictures. Between-subject design. Cognitive testing took place at the University of Basel, Switzerland. Sleep was recorded at participants' homes, using portable electroencephalograph-recording devices. Nine hundred-twenty-nine healthy young participants (mean age 22.48 ± 3.60 y standard deviation). None. In striking contrast to our expectations as well as numerous previous findings, we did not find any significant correlations between sleep and memory consolidation for pictorial stimuli. Our results indicate that individual differences in sleep are much less predictive for pictorial memory processes than previously assumed and suggest that previous studies using small sample sizes might have overestimated the association strength between sleep stage duration and pictorial memory performance. Future studies need to determine whether intraindividual differences rather than interindividual differences in sleep stage duration might be more predictive for the consolidation of emotional and neutral pictures during sleep.


Incidence and Prevalence of Inflammatory Bowel Disease across Asia.
Inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), are chronic inflammatory disorders of the gastrointestinal tract caused by interactions between genetic, environmental, immunological, and microbial factors. While the incidence and prevalence of IBD in Asian populations were relatively lower than those in Western countries, they appear to be gradually increasing. A Westernized diet, high socioeconomic status, improvement of hygiene, and development of vaccination could affect the increases in IBD incidence and prevalence in Asian countries. This review describes the latest trends in the incidence and prevalence of IBD in Asia. Studying the epidemiology of IBD in Asia may unravel the etiopathogenesis of and risk factors for IBD.


Reduction of the small synaptic vesicle protein synaptophysin but not the large dense core chromogranins in the left thalamus of subjects with schizophrenia.
It has been hypothesized that a lesion in the neuronal circuits of thalamus might contribute to the symptoms in schizophrenia. It has also been suggested that impaired synaptic transmission is an important component of the pathophysiology of schizophrenia. In the present study we assess the synaptic integrity of thalamus by means of examining the protein levels of: (1) synaptophysin, a membrane bound protein of small synaptic vesicles, and (2) chromogranins, a family of soluble secretory proteins stored and released from the secretory large dense-core vesicles. The brains of 9 patients with schizophrenia and 9 age-matched control subjects were studied. The levels of synaptophysin and chromogranins were measured by radioimmunoassays. The amount of synaptophysin in the left thalamus was significantly decreased (p = .036) in the schizophrenic group (2655 +/- 605 nmol synaptophysin/mg total protein) compared to the control group (3248 +/- 827 nmol synaptophysin/mg total protein). There were no differences between the groups in the levels of chromogranins, nor in the levels of synaptophysin of the right thalamus. These findings indicate defect synaptic function in the left thalamus of patients with schizophrenia. This may be the cause of a reduction of synaptic terminals or a defect limited to certain structures of the synapse, namely the small presynaptic vesicles.


[Risk factors of dental caries among young children in Pudong New District, Shanghai].
To establish a prospective cohort of kindergarten children and longitudinally study the causes of early childhood caries. Cluster random sampling was used to select a kindergarten in the urban and suburban areas of Pudong New District of Shanghai, a total of 240 small-class children joined the study. Chi-square test, analysis of variance, binomial logistic regression model and general linear regression model was used to analyze caries of the cohort children at baseline and 1 year after follow-up with SPSS 21.0 software package. In the first two years of this cohort study, the follow-up rate was 88.3%, the caries rate in the first year of baseline and follow-up were 58.3% and 69.8%, and the mean dmft values were 3.1±4.2 and 4.5±4.9, respectively. 56.1% of children had new caries. Logistic regression results showed that children who lived in the suburbs (P=0.010) and ate candy more frequently (P=0.036) had higher rates of new caries. The results of general linear regression equation showed that children in the suburbs (P<0.001), those who did not use fluoridated toothpaste (P=0.003) and those who ate candy more frequently (P=0.002) had higher new mean dmft values. Living in the suburbs, not using fluoride toothpaste and eating candy more frequently are important risk factors for new caries in preschool children in Pudong New District of Shanghai.


Influence of maternal smoking on variability of peak expiratory flow rate in school children.
Diurnal variability of peak expiratory flow rates (PEFRs) was assessed in 1,237 children. The PEFR was measured twice daily over a 1-week period. As an index of variability, the log of a week's mean of daily amplitude was calculated. Linear regression analyses revealed a significant positive association between maternal smoking and the variability of PEFR for nonasthmatic children. For these children, exposure to maternal smoking was associated with a 13.7 percent increase (confidence interval [CI], 3.8 to 24.7 percent) in PEFR variability. For asthmatic children an effect was found for nonatopic (54.7 percent increase; CI, 5.5 to 226.8 percent) but not for atopic children (-8.5 percent change; CI, -41.2 to 42.3 percent). In the latter group, there was evidence that mothers changed their smoking habits subsequent to the development of disease in their children. We conclude that exposure to maternal smoking can increase the variability of PEFR and thus might contribute to the development of asthma.


NF-kappaB activation upregulates fibroblast growth factor 8 expression in prostate cancer cells.
Fibroblast growth factor 8 (FGF8) is over-expressed in prostate cancer (CaP) correlating with high-grade disease and reduced survival. The role of acetylation in transcriptional regulation of FGF8 was investigated using the histone deacetylase (HDAC) inhibitor Trichostatin A (TSA). FGF8 transcriptional response to TSA was investigated by gene reporter assays, RT-PCR, and Western blotting. Chromatin immunoprecipitation (ChIP) assays were also performed. FGF8 is upregulated in response to TSA treatment along with NF-kappaB transcriptional activity. Over-expression of p65 activated FGF8 transcription. ChIP assays revealed p65 recruitment to the fgf8 promoter, containing putative NF-kappaB binding sites, post TSA stimulation. PI-3K activity is required for TSA mediated FGF8 upregulation. Using TSA treatment in prostate cancer cells, a requirement of PI-3K activity in mediating TSA function is demonstrated and a novel role for NF-kappaB in the regulation of FGF8 expression is uncovered.


Contingent leadership and effectiveness of trauma resuscitation teams.
This research investigated leadership and effectiveness of teams operating in a high-velocity environment, specifically trauma resuscitation teams. On the basis of the literature and their own ethnographic work, the authors proposed and tested a contingency model in which the influence of leadership on team effectiveness during trauma resuscitation differs according to the situation. Results indicated that empowering leadership was more effective when trauma severity was low and when team experience was high. Directive leadership was more effective when trauma severity was high or when the team was inexperienced. Findings also suggested that an empowering leader provided more learning opportunities than did a directive leader. The major contribution of this article is the linkage of leadership to team effectiveness, as moderated by relatively specific situational contingencies.


Efficient lipid staining in plant material with sudan red 7B or fluorol [correction of fluoral] yellow 088 in polyethylene glycol-glycerol.
Polyethylene glycol (400) with 90% glycerol (aqueous) is introduced as an efficient solvent system for lipid stains. Various lipid-soluble dyes were dissolved in this solvent system and tested for their intensity, contrast, and specificity of staining of suberin lamellae in plant tissue. The stability (i.e., lack of precipitation) of the various staining solutions in the presence of fresh tissue was also tested. When dissolved in polyethylene glycol-glycerol, Sudan red 7B (fat red) was the best nonfluorescent stain and fluorol yellow 088 (solvent green 4) was an excellent fluorochrome. These two dyes formed stable staining solutions which efficiently stained lipids in fresh sections without forming precipitates. Estimations of the solubilities of these dyes in the solvent compared with their solubilities in lipids of various chemical types indicated that they should both be effective stains for lipids in general.


Characterization of purinergic P2X4 receptor channels expressed in anterior pituitary cells.
Anterior pituitary cells express cation-conducting P2X receptor channels (P2XRs), but their molecular identity, electrophysiological properties, cell-specific expression pattern, and physiological roles have been only partially characterized. In this study, we show by quantitative RT-PCR that mRNA transcripts for the P2X(4) subunit are the most abundant in rat anterior pituitary tissue and confirm the P2X(4)R protein expression by Western blot analysis. Single-cell patch-clamp recordings show that extracellular ATP induced an inward depolarizing current in a majority of thyrotropin-releasing hormone-responsive pituitary cells, which resembled the current profile generated by recombinant P2X(4)R. The channels were activated and desensitized in a dose-dependent manner and deactivated rapidly. Activation of these channels led to stimulation of electrical activity and promotion of voltage-gated and voltage-insensitive Ca(2+) influx. In the presence of ivermectin, a specific allosteric modulator of P2X(4)Rs, there was an approximately fourfold increase in the maximum amplitude of the ATP-induced inward current, accompanied by an increase in the sensitivity of receptors for ATP, slowed deactivation of receptors, and enhanced ATP-induced prolactin release. These results indicate that thyrotropin-releasing hormone-responsive cells, including lactotrophs, express homomeric and/or heteromeric P2X(4)Rs, which facilitate Ca(2+) influx and hormone secretion.


Individual differences in the peripheral immune system promote resilience versus susceptibility to social stress.
Depression and anxiety disorders are associated with increased release of peripheral cytokines; however, their functional relevance remains unknown. Using a social stress model in mice, we find preexisting individual differences in the sensitivity of the peripheral immune system that predict and promote vulnerability to social stress. Cytokine profiles were obtained 20 min after the first social stress exposure. Of the cytokines regulated by stress, IL-6 was most highly up-regulated only in mice that ultimately developed a susceptible behavioral phenotype following a subsequent chronic stress, and levels remained elevated for at least 1 mo. We confirmed a similar elevation of serum IL-6 in two separate cohorts of patients with treatment-resistant major depressive disorder. Before any physical contact in mice, we observed individual differences in IL-6 levels from ex vivo stimulated leukocytes that predict susceptibility versus resilience to a subsequent stressor. To shift the sensitivity of the peripheral immune system to a pro- or antidepressant state, bone marrow (BM) chimeras were generated by transplanting hematopoietic progenitor cells from stress-susceptible mice releasing high IL-6 or from IL-6 knockout (IL-6(-/-)) mice. Stress-susceptible BM chimeras exhibited increased social avoidance behavior after exposure to either subthreshold repeated social defeat stress (RSDS) or a purely emotional stressor termed witness defeat. IL-6(-/-) BM chimeric and IL-6(-/-) mice, as well as those treated with a systemic IL-6 monoclonal antibody, were resilient to social stress. These data establish that preexisting differences in stress-responsive IL-6 release from BM-derived leukocytes functionally contribute to social stress-induced behavioral abnormalities.


The island pedicle rotation advancement gastrocnemius musculocutaneous flap for complete coverage of the popliteal fossa.
The medial head of the gastrocnemius flap is a large, versatile flap that can be used to cover areas above the popliteal fossa, thereby minimizing the necessity for amputation in many traumatic injuries to this area. By advancing the medial head of the gastrocnemius muscle and rotating it on the neurovascular pedicle of the medial sural vessels, we have covered a 17 cm X 20 cm defect of crushed muscle, tendon, and bone in a leg that was traumatically amputated proximal to the knee. This procedure makes possible muscular coverage for bone, stainless steel plate, and reanastomosed arteries and veins above the knee and provides skin coverage extending to 70% of the circumference of the popliteal fossa. Autografting provides coverage for the remainder of the wound. Subsequent extension and flexion of the viable knee joint was achieved.


Conformational states of human purine nucleoside phosphorylase at rest, at work, and with transition state analogues.
Human purine nucleoside phosphorylase (PNP) is a homotrimer binding tightly to the transition state analogues Immucillin-H (ImmH; K(d) = 56 pM) and DATMe-ImmH-Immucillin-H (DATMe-ImmH; K(d) = 8.6 pM). ImmH binds with a larger entropic penalty than DATMe-ImmH, a chemically more flexible inhibitor. The testable hypothesis is that PNP conformational states are more relaxed (dynamic) with DATMe-ImmH, despite tighter binding than with ImmH. PNP conformations are probed by peptide amide deuterium exchange (HDX) using liquid chromatography high-resolution Fourier transform ion cyclotron resonance mass spectrometry and by sedimentation rates. Catalytically equilibrating Michaelis complexes (PNP.PO(4).inosine <--> PNP.Hx.R-1-P) and inhibited complexes (PNP.PO(4).DATMe-ImmH and PNP.PO(4).ImmH) show protection from HDX at 9, 13, and 15 sites per subunit relative to resting PNP (PNP.PO(4)) in extended incubations. The PNP.PO(4).ImmH complex is more compact (by sedimentation rate) than the other complexes. HDX kinetic analysis of ligand-protected sites corresponds to peptides near the catalytic sites. HDX and sedimentation results establish that PNP protein conformation (dynamic motion) correlates more closely with entropy of binding than with affinity. Catalytically active turnover with saturated substrate sites causes less change in HDX and sedimentation rates than binding of transition state analogues. DATMe-ImmH more closely mimics the transition of human PNP than does ImmH and achieves strong binding interactions at the catalytic site while causing relatively modest alterations of the protein dynamic motion. Transition state analogues causing the most rigid, closed protein conformation are therefore not necessarily the most tightly bound. Close mimics of the transition state are hypothesized to retain enzymatic dynamic motions related to transition state formation.


Vasorin deficiency leads to cardiac hypertrophy by targeting MYL7 in young mice.
Vasorin (VASN) is an important transmembrane protein associated with development and disease. However, it is not clear whether the death of mice with VASN deficiency (VASN-/- ) is related to cardiac dysfunction. The aim of this research was to ascertain whether VASN induces pathological cardiac hypertrophy by targeting myosin light chain 7 (MYL7). VASN-/- mice were produced by CRISPR/Cas9 technology and inbreeding. PCR amplification, electrophoresis, real-time PCR and Western blotting were used to confirm VASN deficiency. Cardiac hypertrophy was examined by blood tests, histological analysis and real-time PCR, and key downstream factors were identified by RNA sequencing and real-time PCR. Western blotting, immunohistochemistry and electron microscopy analysis were used to confirm the downregulation of MYL7 production and cardiac structural changes. Our results showed that sudden death of VASN-/- mice occurred 21-28 days after birth. The obvious increases in cardiovascular risk, heart weight and myocardial volume and the upregulation of hypertrophy marker gene expression indicated that cardiac hypertrophy may be the cause of death in young VASN-/- mice. Transcriptome analysis revealed that VASN deficiency led to MYL7 downregulation, which induced myocardial structure abnormalities and disorders. Our results revealed a pathological phenomenon in which VASN deficiency may lead to cardiac hypertrophy by downregulating MYL7 production. However, more research is necessary to elucidate the underlying mechanism.


Determinants of Organic Fertilizers Utilization Among Smallholder Farmers in South Gondar Zone, Ethiopia.
Adoption of organic fertilization is low among farmers in rural areas of Ethiopia, affecting yields and general food security in the region. This study aimed to identify the determinants of the utilization of organic fertilizers among smallholder farmers in the South Gondar Zone, Amhara National Regional State (ANRS), Northwest Ethiopia. A community-based cross-sectional study was used among smallholder farmers in the South Gondar Zone, ANRS, Northwest Ethiopia. Primary data were collected from 420 sample respondents using multistage sampling with a combination of both simple random and cluster sampling techniques. The binary logistic regression model was used to assess the use of organic fertilizers among smallholder farmers in the South Gondar Zone. The results are presented as adjusted odds ratios (AOR) together with their corresponding 95% confidence intervals. head of household age (AOR = 1.099, 95% CI 1.018-1.187), married marital status (AOR = 10.506, 95% CI 1.355-81.427), literate head of household (AOR = 3.323, 95% CI 1.571-7.029), number of laborers (AOR = 1.442, 95% CI 1.060-1.962), farming experience (AOR = 1.132, 95% CI 1.041-1.232), farm size (AOR = 1.063, 95% CI 1.008-1.121), and number of livestock (AOR = 1.368, 95% CI 1.115-1.677) were positively associated with the utilization of organic fertilizer while single marital status (AOR = 0.062, 95% CI 0.004-0.851), cost of laborer (AOR = 0.965, 95% CI 0.951-0.978), household income (AOR = 0.880, 95% CI 0.824-0.939), medium soil fertility (AOR = 0.039, 95% CI 0.007-0.229), fertile soil (AOR = 0.020, 95% CI 0.003-0.120), and home to farm distance (AOR = 0.219, 95% CI 0.067-0.717) were negatively associated with the utilization of organic fertilizer. This finding showed that multiple variables have an effect to determining the use of organic fertilizer by smallholder farmers. Therefore, the finding is important to adopt programs to encourage the use of organic fertilizer, implement policies in an attempt to adapt the use of organic fertilizer among the South Gondar Zone, and critically consider these factors. Furthermore, extension workers should focus on raising awareness about the importance of organic fertilizers to encourage or expand their use among smallholder farmers.


Sensitive arginine sensing based on inner filter effect of Au nanoparticles on the fluorescence of CdTe quantum dots.
Arginine plays an important role in many biological functions, whose detection is very significant. Herein, a sensitive, simple and cost-effective fluorescent method for the detection of arginine has been developed based on the inner filter effect (IFE) of citrate-stabilized gold nanoparticles (AuNPs) on the fluorescence of thioglycolic acid-capped CdTe quantum dots (QDs). When citrate-stabilized AuNPs were mixed with thioglycolic acid-capped CdTe QDs, the fluorescence of CdTe QDs was significantly quenched by AuNPs via the IFE. With the presence of arginine, arginine could induce the aggregation and corresponding absorption spectra change of AuNPs, which then IFE-decreased fluorescence could gradually recover with increasing amounts of arginine, achieving fluorescence "turn on" sensing for arginine. The detection mechanism is clearly illustrated and various experimental conditions were also optimized. Under the optimum conditions, a decent linear relationship was obtained in the range from 16 to 121μgL-1 and the limit of detection was 5.6μgL-1. And satisfactory results were achieved in arginine analysis using arginine injection, compound amino acid injection, even blood plasma as samples. Therefore, the present assay showed various merits, such as simplicity, low cost, high sensitivity and selectivity, making it promising for sensing arginine in biological samples.


Glow-peak stability in 6LiF:Mg,Ti (TLD-600) exposed to a Fe-ion beam.
The stability of glow peaks in 6LiF:Mg,Ti (TLD-600) exposed to a high-energy Fe-ion beam was examined in comparison to 137Cs gamma-ray irradiation under changing annealing conditions. The peak areas induced by the Fe ions were much smaller than those by gamma-rays. The sizes and positions of peaks 3-5 in Fe-ion irradiated samples were hardly changed after post-annealing at 100 degrees C x 30 min, regardless of the pre-annealing conditions (fast quenching or subsequent pre-annealing at 100 degrees C x 2 h). Whereas, the peaks in gamma-ray irradiated samples were notably affected by post-annealing; the peak positions and peak-area sizes changed in different ways depending on the pre-annealing conditions. The effects of post-annealing on peak 6 were identical for Fe ions and gamma-rays. These facts suggest that peaks 3-5 in TLD-600 comprised both stable and unstable luminescent centers, and that the latter part would be easily depleted in highly dense ionization.


Centrilobular distribution of acetaldehyde and collagen in the ethanol-fed micropig.
We established a new animal model of alcoholic liver disease in the micropig, a species that consumes ethanol voluntarily in the diet. Ten micropigs were pair-fed diets containing 40% of calories as ethanol or cornstarch with identical amounts of fat, protein and micronutrients for 12 mo. Liver histopathology in the ethanol-fed pigs included steatonecrosis in all five and interstitial and perivenous fibrosis in three. Electron microscopy showed Ito-cell transformation with perisinusoidal collagen accumulation. Acetaldehyde adducts were found by immunofluorescence in the centrilobular region and were focused in perivenous zone 3 of all ethanol-fed animals. Protein and triglyceride levels were increased, whereas vitamin A and iron levels were decreased in liver homogenates from ethanol-fed animals. Thus, in this new animal model of alcoholism, ethanol feeding produced the features of alcoholic liver disease concurrent with hepatic deficiency of selected nutrients. Histological and immunofluorescent studies provide in vivo evidence that perivenous collagen deposition is linked to ethanol metabolism and acetaldehyde production.


Effect of dose on the disposition of 2-ethoxyethanol after inhalation by F344/N rats.
Glycol ethers such as 2-ethoxyethanol (EE) are widely used as solvents because they are miscible in aqueous and organic solutions. Toxic effects of EE in rodents include teratogenicity, fetotoxicity, hematotoxicity, and testicular atrophy. The purpose of this study was to determine the effect of dose on the absorption, metabolism, and excretion of 2-ethoxy [U-14C]ethanol by F344/N rats after inhalation exposure. Rats were exposed to either 5 ppm EE for 5 hr 40 min or 46 ppm EE for 6 hr. The uptake and metabolism of EE were linear in the concentration range studied. Significant percentages of the retained doses were exhaled during (22%) and after exposure (16%) as 14CO2. Forty-six percent of the retained dose was excreted in the urine. Approximately 10% of the retained dose was detected in the carcass 66 hr after exposure. The major urinary metabolite was ethoxyacetic acid (EAA), the toxic metabolite of EE. The amount of EAA excreted was linearly related to exposure concentration. Ethylene glycol and N-ethoxyacetyl glycinate were identified as minor metabolites excreted in the urine. The results of this study suggest that the toxicity of inhaled EE should be directly proportional to the exposure concentration up to 46 ppm if the toxicity of EE is due to EAA.


Reestablishment of hope as an intervention for a patient with cystic fibrosis awaiting lung transplantation.
Patients with cystic fibrosis (CF) experience a progressive deterioration in health leading to a shortened life expectancy with concomitant psychologic challenges that also may require intervention. Approaches to address psychologic dysfunction include insight-oriented therapy, cognitive-behavioral therapy, biofeedback, hypnosis, massage, music therapy, relaxation, and family therapy. Patients who learn to use coping strategies are more likely to adhere to treatment, which helps improve both their physical and psychologic well-being. Unique stressors arise for patients awaiting lung transplant as a result of their advanced state of disease along with their fear of death before donor lungs become available. This case report demonstrates some of the psychologic struggles faced by a 10-year-old with CF as he awaited lung transplantation, which led to a loss of hope and his request for withdrawal of care. Encouraging of the patient to express how his life might be improved yielded changes in his management that restored his hopefulness, and allowed him to survive long enough to undergo successful lung transplantation. Thus, reestablishment of hope might be studied as an additional intervention to help improve the well-being of patients who are seriously ill.


Prediction of protein secondary structure content.
All existing algorithms for predicting the content of protein secondary structure elements have been based on the conventional amino-acid-composition, where no sequence coupling effects are taken into account. In this article, an algorithm was developed for predicting the content of protein secondary structure elements that was based on a new amino-acid-composition, in which the sequence coupling effects are explicitly included through a series of conditional probability elements. The prediction was examined by a self-consistency test and an independent dataset test. Both indicated a remarkable improvement obtained when using the current algorithm to predict the contents of alpha-helix, beta-sheet, beta-bridge, 3(10)-helix, pi-helix, H-bonded turn, bend and random coil. Examples of the improved accuracy by introducing the new amino-acid-composition, as well as its impact on the study of protein structural class and biologically function, are discussed.


Learning for clinical leadership.
Clinical leadership has been acclaimed widely as a major factor influencing the quality of patient care but research has revealed a paucity of preparation for this significant role. Leadership literature has rarely addressed clinical leadership specifically or referred to the difficulties in characterizing effective clinical leaders. The research informing this paper focused on clinical leadership and identified five attributes of effective clinical leaders: creativity, highlighting, influencing, respecting, and supporting. Effective clinical leaders adopted a transformational leadership style and improved care, through others, by including transformational (soft) knowledge as an integral part of their effective practice repertoire. Phronesis is introduced as practical wisdom that is gained through immersion in relevant experience, and as an essential element of preparation for clinical nursing leadership practice. It is argued, that learning to transform care requires opportunities to work within an environment that engenders and supports aspiring leaders. The paper describes the research process, elucidates the attributes through illustrative examples from the research data, and discusses an emergent educational strategy for the development of these attributes by clinicians in their practice environments. The paper also describes the application of this research through an interdisciplinary programme for staff leading teams in both health and social services sectors.


Effect of nivazol in Nelson's syndrome.
We report the use of nivazol in a patient with Nelson's syndrome. Nivazol was highly effective in reducing ACTH secretion but, contrary to reports of its use in other primates, in our patient nivazol did have systemic glucocorticoid effects.


The identification of a tetracycline resistance gene tet(M), on a Tn916-like transposon, in the Bacillus cereus group.
In order to investigate whether resistance genes present in bacteria in manure could transfer to indigenous soil bacteria, resistant isolates belonging to the Bacillus cereus group (Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis) were isolated from farm soil (72 isolates) and manure (12 isolates) samples. These isolates were screened for tetracycline resistance genes (tet(K), tet(L), tet(M), tet(O), tet(S) and tet(T)). Of 88 isolates examined, three (3.4%) isolates carried both tet(M) and tet(L) genes, while four (4.5%) isolates carried the tet(L) gene. Eighty-one (92.1%) isolates did not contain any of the tested genes. All tet(M) positive isolates carried transposon Tn916 and could transfer this mobile DNA element to other Gram-positive bacteria.


[Differential gene expression in the wear particle induced and infectious periprosthetic membrane of loosened knee-endoprostheses].
About 5 to 12 % of hip endoprostheses will loosen after ten years. The periprosthetic membran between bone and prosthesis plays a crucial role in prosthesis loosening. Different pathomechanisms lead to the growth of such a membran, which can be discriminated by different histomorphologies: wear particle induced type, infectious type, combined type, indifferent type. 8 hybridizations were performed on PIQOR cDNA arrays. Objects of the study were periprosthetic interface tissue samples from 3 patients with particle induced and 2 patients with infectious prosthesis loosening. Tissue parts directly adjacent to the site of RNA-isolation were analyzed immuno-/ histopathologically in order to overcome the problem of tissue heterogeneity. 34 genes were found constantly differentially expressed, among which were cd9, cd11b, cd18, cd68, osteopontin, ferritin heavy-chain upregulated in the particle induced membrane and collagen types 1alpha-1, 3alpha-1, integrin alpha-1, thrombospondin 2 and nidogen upregulated in the infectious membrane. The most striking finding was the strong upregulation (from 20 fold to 323 fold) of megakaryocyte stimulating factor (msf) in all wear particle cases and 1 out of 2 infectious cases, which was confirmed by real-time RT-PCR. The upregulation of msf suggests an important pathogenetic role: The msf splice product lubricin is responsible for the lubrification of healthy joints, but its excellent lubrification ability may disturb the tight interaction between bone and prosthesis and thereby contribute to prosthesis loosening.


Diabetes epidemic sweeping the Arab world.
The prevalence of type-2 diabetes mellitus (T2DM) has increased dramatically during the last 2 decades, a fact driven by the increased prevalence of obesity, the primary risk factor for T2DM. The figures for diabetes in the Arab world are particularly startling as the number of people with diabetes is projected to increase by 96.2% by 2035. Genetic risk factors may play a crucial role in this uncontrolled raise in the prevalence of T2DM in the Middle Eastern region. However, factors such as obesity, rapid urbanization and lack of exercise are other key determinants of this rapid increase in the rate of T2DM in the Arab world. The unavailability of an effective program to defeat T2DM has serious consequences on the increasing rise of this disease, where available data indicates an unusually high prevalence of T2DM in Arabian children less than 18 years old. Living with T2DM is problematic as well, since T2DM has become the 5(th) leading cause of disability, which was ranked 10(th) as recently as 1990. Giving the current status of T2DM in the Arab world, a collaborative international effort is needed for fighting further spread of this disease.


Assessment of the Road Surface Condition with Longitudinal Acceleration Signal of the Car Body.
On the basis of road tests, the authors assessed the feasibility of the vehicle body acceleration values for the purposes of assessing road surface characteristics in terms of its roughness. Short-term Fourier Transform (STFT) was used for the analysis of the recorded signal. The spectra obtained in successive frequency bands demonstrate the amplitudes originating from the natural vibrations of the rolling wheel and forces resulting from the interaction with the road roughness. The article focuses on the relationships between the road roughness and the ratios of individual amplitudes in a specific frequency band of the vehicle body acceleration values. Amplitude values derived on the basis of successive windows were averaged for analogous, arbitrarily assumed local frequency bands. The value characterizing the road surface condition provided the information regarding the mean amplitude value in specific frequency ranges depending on the instantaneous velocity of the car body and the condition of the road surface on which it was moving. In cases where the road was free of any visible roughness, the obtained mean amplitude value in the analyzed spectrum window, for the adopted vehicle velocity range from 50 km h to 100 km/h, did not exceed 0.02 m/s2. It was also demonstrated that the road surface roughness leads to an increase in the mean amplitude value from 0.07 m/s2 to 0.16 m/s2.


Extra-anatomic bypass: a new look (opposing view).
The criteria by which revascularizations are measured ultimately is patency, since cessation of blood flow equates with a failed graft and usually signals a return to the preoperative limb perfusion or less. An uncommonly analyzed criterion for graft function is blood flow capacity through the reconstruction which is critically important when increased blood flow demands are required with exercise. This is particularly true in the aortoiliac segment when the bypass must meet large blood flow requirements. When the extra-anatomic bypass is evaluated in this manner, it is often inadequate to meet the blood flow demands required with activity. The source of blood flow is the subclavian artery and not the aorta. The graft diameter is almost one half that used to bypass or substitute for the aorta and four times as long. It is not surprising, therefore, that increases in blood flow are limited in this remote subcutaneous bypass circuit. The axillofemoral and, to some extent, the femorfemoral bypass grafts produce hemodynamic gradients to blood flow increases above basal rates. Thus, graft patency alone does not realistically measure the functional capacity of an aortoiliac reconstruction. The conceptual design of an extra-anatomic bypass results in hemodynamic liabilities that produce an inferior performance to that observed with in-line direct aortofemoral bypass. This observation further supports our contention that extra-anatomic bypass of the aortoiliac segment should be restricted to circumvent a prosthetic graft infection in the aortoiliac segment to maintain limb perfusion following graft removal. Whether it has a role in selected patients with a short life span and critical ischemia who appear to be a prohibited risk for in-line aortofemoral reconstruction will always be debatable. Current experience does not justify expanded indications for extra-anatomic bypass in symptomatic patients with aortoiliac disease. The proven effective and durable intervention is aortofemoral revascularization.


Characterization of CRISPR-Cas system in clinical Staphylococcus epidermidis strains revealed its potential association with bacterial infection sites.
Staphylococcus epidermidis is considered as a major cause of nosocomial infections, bringing an immense burden to healthcare systems. Virulent phages have been confirmed to be efficient in combating the pathogen, but the prensence of CRISPR-Cas system, which is a bacterial immune system eliminating phages was reported in few S. epidermidis strains. In this study, the CRISPR-Cas system was detected in 12 from almost 300 published genomes in GenBank and by PCR of cas6 gene in 18 strains out of 130 clinical isolates obtained in Copenhagen. Four strains isolated in 1965-1966 harboured CRISPR elements confirming that this immunity system was not recently acquired by S. epidermidis. In these CRISPR-positive strains, 44 and 12 spacers were found to belong to CRISPR1 and CRISPR2 elements, respectively. However, only 15 spacers displayed homology to reported phages and plasmids DNA. Interestingly, 5 different spacers located in the CRISPR1 locus with homolgy to virulent phage 6ec DNA sequences, and 19 strains each carrying 2 or 3 different spacers recognizing this phage, implied that the CRISPR-Cas immunity could be abrogated by nucleotide mismatch between the spacer and its target phage sequence, while new spacers obtained from the evolved phage could recover the CRISPR interference. In addition, phylogenetic analysis of the 29 CRISPR-positive isolates divided them into four lineages, with 81% human blood isolates as a distinct sub-lineage, suggesting that the CRISPR difference is closely related to diverse habitats. Knowledge of CRISPR and its prevalence may ultimately be applied in the understanding of origin and evolution of CRISPR-positive S. epidermidis strains.


Antineoplastic activities of MT81 and its structural analogue in Ehrlich ascites carcinoma-bearing Swiss Albino mice.
Many fungal toxins exhibit in vitro and in vivo antineoplastic effects on various cancer cell types. Luteoskyrin,a hydroxyanthraquinone has been proved to be a potent inhibitor against Ehrlich ascites tumor cells. The comparative antitumor activity and antioxidant status of MT81 and its structural analogue [Acetic acid-MT81 (Aa-MT81)] having polyhydroxyanthraquinone structure were assessed against Ehrlich ascites carcinoma (EAC) tumor in mice. The in vitro cytotoxicity was measured by the viability of EAC cells after direct treatment of the said compounds. In in vivo study, MT81 and its structural analogue were administered (i.p.) at the two different doses (5, 7 mg MT81; 8.93, 11.48 mg Aa-MT81/kg body weight) for 7 days after 24 hrs. of tumor inoculation. The activities were assessed using mean survival time (MST), increased life span (ILS), tumor volume, viable tumor cell count, peritoneal cell count, protein percentage and hematological parameters. Antioxidant status was determined by malondialdehyde (MDA) and reduced glutathione (GSH) content, and by the activity of superoxide dismutase (SOD) and catalase (CAT). MT81 and its structural analogues increased the mean survival time, normal peritoneal cell count. They decreased the tumor volume, viable tumor cell count, hemoglobin percentage and packed cell volume. Differential counts of WBC, total counts of RBC & WBC that altered by EAC inoculation, were restored in a dose-dependent manner. Increased MDA and decreased GSH content and reduced activity of SOD, and catalase in EAC bearing mice were returned towards normal after the treatment of MT81 and its structural analogue. Being less toxic than parent toxin MT81, the structural analogue showed more prominent antineoplastic activities against EAC cells compared to MT81. At the same time, both compounds exhibit to some extent antioxidant potential for the EAC-bearing mice.


Comparison of Neonatal Abstinence Syndrome Manifestations in Preterm Versus Term Opioid-Exposed Infants.
Twenty percent to 40% of infants exposed to in utero opioid were delivered preterm. There is currently no neonatal abstinence syndrome (NAS) scoring tool known to accurately evaluate preterm opioid-exposed infants. This can lead to difficulties in titrating pharmacotherapy in this population. To describe NAS symptoms in preterm opioid-exposed infants in comparison with matched full-term controls. This was a retrospective cohort study from a single tertiary care center of methadone-exposed infants born between 2006 and 2010. Using modified Finnegan scale scores recorded every 3 to 4 hours beginning at 6 hours of life until 24 to 48 hours after medication discontinuation, NAS symptoms was compared between 45 preterm infants and 49 full-term matched controls. Concurrent neonatal medical diagnoses were also compared. The median gestational age in the preterm group was 35 weeks (interquartile range [IQR] = 33-36) versus 39 weeks (IQR = 38-40) in the term group. Preterm infants scored less frequently for many items including sleep disturbance (24.4% vs 46.2%), tremors (77.9% vs 89.7%), muscle tone (87.9% vs 97.4%), sweating (2.1% vs 9.4%), nasal stuffiness (11.9% vs 20.5%), and loose stools (7.0% vs 14.3%) than full-term controls. Preterm infants scored more frequently for hyperactive moro reflex (26.4% vs 5.5%), tachypnea (19.3% vs 16.1%), and poor feeding (24.6% vs 11.8%). Provider awareness of differences in manifestations of preterm and term infants with NAS, as well as concurrent prematurity diagnoses that can influence NAS scoring, is needed. These findings mandate the development of a modified NAS scoring tool for the preterm NAS population. A preterm NAS scoring tool needs to be developed and validated to more accurately evaluate and treat preterm opioid-exposed infants.


MDM2 gene promoter polymorphisms and risk of lung cancer: a case-control analysis.
The MDM2 protein negatively regulates p53 expression level in modulating DNA repair, cell-cycle control, cell growth and apoptosis. Polymorphisms in the promoter region of the MDM2 gene have been shown to alter protein expression and may, thus, play a role in carcinogenesis. To test our hypothesis that the MDM2 promoter polymorphisms are associated with risk of lung cancer, we conducted a hospital-based, case-control study of 1026 non-Hispanic white patients newly diagnosed with lung cancer and 1145 cancer-free controls who were frequency-matched by age (+/-5 years), sex, ethnicity and smoking status. We genotyped for the MDM2 promoter G2580T (also called SNP309) and G2164A polymorphisms that have a minor allele frequency >0.05. The distributions of the MDM2-2580G variant allele and genotypes were significantly less common among the cases than among the controls (P = 0.038 and 0.045, respectively), but this was not evident for MDM2-2164G (P = 0.865 and 0.614, respectively). Compared with the MDM2-2580TT genotype, the MDM2-2580G variant genotypes were associated with a decreased risk of lung cancer [odds ratio = 0.81 and 95% confidence interval = 0.67-0.98 for GT, 0.83 (0.63-1.08) for GG, and 0.81 (0.68-0.97) for the combined GT/GG genotype]. However, no significant association was observed between the MDM2-2164G variant genotypes and lung cancer risk. Our results suggest that the MDM2-2580G allele may be a marker of reduced genetic susceptibility to lung cancer in the non-Hispanic white population, a finding that seems to contradict previous reports.


Interference of plant volatiles on pheromone receptor neurons of male Grapholita molesta (Lepidoptera: Tortricidae).
In moths, sex pheromone components are detected by pheromone-specific olfactory receptor neurons (ph-ORNs) housed in sensilla trichodea in the male antennae. In Grapholita molesta, ph-ORNs are highly sensitive and specific to the individual sex pheromone components, and thus help in the detection and discrimination of the unique conspecific pheromone blend. Plant odors interspersed with a sub-optimal pheromone dose are reported to increase male moth attraction. To determine if the behavioral synergism of pheromone and plant odors starts at the ph-ORN level, single sensillum recordings were performed on Z8-12:Ac and E8-12:Ac ph-ORNs (Z-ORNs and E-ORNs, respectively) stimulated with pheromone-plant volatile mixtures. First, biologically meaningful plant-volatile doses were determined by recording the response of plant-specific ORNs housed in sensilla auricillica and trichodea to several plant odorants. This exploration provided a first glance at plant ORNs in this species. Then, using these plant volatile doses, we found that the spontaneous activity of ph-ORNs was not affected by the stimulation with plant volatiles, but that a binary mixture of sex pheromone and plant odorants resulted in a small (about 15%), dose-independent, but statistically significant, reduction in the spike frequency of Z-ORNs with respect to stimulation with Z8-12:Ac alone. The response of E-ORNs to a combination of E8-12:Ac and plant volatiles was not different from E8-12:Ac alone. We argue that the small inhibition of Z-ORNs caused by physiologically realistic plant volatile doses is probably not fully responsible for the observed behavioral synergism of pheromone and plant odors.