Conflicts in ethical problems of patient education: strategies for hypertension control explore contractual approach.
Experiences in the training of health education interviewers for an intervention strategy in a study which utilized health education strategies for control of hypertension afforded the opportunity for examining the relationship of research to practice and for addressing a recurring concern of health educators in practice, i.e., concern over the ethics of adopting a directing role with patients. This concern, which results in ambivalence in presentation and extreme postures of authoritativeness or passivity as educators, detracts from the potential of a health education intervention. While not evaluated comparatively with other strategies as to outcome, one method of addressing this problem, namely explication of the contractual approach in health education, was utilized informally and has implications for health education interventions. This approach utilizes the patient as a truly active partner and sharpens the use of the educational diagnosis. The outcome has potential positives for patients in personal growth and for educators in the sharpening of skills and in more effective use of their resources. These observations and the ensuing discussions come as an unplanned output of the study from which they derived. However, a variety of experiences in health education lend weight to the impression that the concerns addressed are not atypical of universal concerns among practitioners. Such conclusions suggest the timeliness of testing out the approach in an experimental fashion in an effort to document its use as technique as well as its effectiveness in ultimate outcome.


Exploring Diener's Multidimensional Conceptualization of Well-Being Through Network Psychometrics.
Previous research has addressed the study of the well-being structure focusing on the separability among hedonic and eudaimonic dimensions by means of latent variable approaches. Diener's tripartite model of Subjective Well-Being has often been identified with hedonic well-being, while flourishing (i.e., positive functioning) has been classified as a proxy for eudaimonic well-being. Instead of pursuing a theory-driven approach, the structure of well-being dimensions can be retrieved by means of network psychometrics that is a highly informative, data-driven approach that allows the model structure to spontaneously emerge from the relationships among indicators. Furthermore, we propose a strength centrality decomposition that is able to summarize the influence of a specific indicator within its dimension and between the other dimensions. Contrary to the theoretical conjecture that the separability among well-being dimensions can be grounded on a hedonic and eudaimonic distinction, our results point to a categorization of the Diener's well-being indicators based on the type of information they convey, that is global evaluation of life satisfaction, positive and negative affective experiences, and perceived positive functioning in life.


[Retinotopic organization of the lateral suprasylvian sulcus' posterior-medial area as revealed by analysis of the pattern of cortico-cortical connection with the cat 17th area].
Using cortico-cortical connection analysis technique, the cat visual area PMLS (the area located on posterior medial wall of the lateral suprasylvian sulcus) retinotopic organization was investigated. A retrograde axonal tracer: horseradish peroxidase (HRP), was injected in the PMLS, and initial neurons were investigated in area 17. It was shown that after HRP injection in PMLS locus, where a central vision field is located, a labelled cell pattern in area 17 corresponded to the L. Palmer et al., 1978, retinotopic map. On the contrary, after HRP injection in PMLS locus, where an upper vision field must be located, as L. Palmer et al., 1978 predicted, initial neurons are visualized in area 17 loci where low visual periphery is displayed: -10 degrees to -60 degrees in vertical meridian and 40 degrees to 80 degrees in horizontal meridian. Such discrepancy in upper and lower visual field representation was also obtained in electrophysiological and topographic investigations by Grant, Shipp, 1991. The data suggest necessity of S. Grant and S. Shipp's retinotopic map use in the cat area PMLS morphofunctional investigation.


Dynamics of pump-induced refractive index changes in single-mode Yb-doped optical fibers.
We quantify the refractive index changes (RIC) in single-mode ytterbium-doped optical fibers induced by optical pulses at 980 nm and, for the first time, report details of the effect dynamics. The RIC dynamics is shown to follow that of the population of the excited/unexcited ion states with a factor proportional to their polarizability difference (PD). The absolute PD value is evaluated in the spectral range of 1460-1620 nm for different fiber samples and is found to be independent on the fiber geometry and on the ion concentration. The PD dispersion profile indicates to a predominant far-resonance UV rather than near-resonant IR transitions contribution to the RIC.


Effectiveness of Start to Run, a 6-week training program for novice runners, on increasing health-enhancing physical activity: a controlled study.
The use of the organized sports sector as a setting for health-promotion is a relatively new strategy. In the past few years, different countries have been investing resources in the organized sports sector for promoting health-enhancing physical activity. In the Netherlands, National Sports Federations were funded to develop and implement "easily accessible" sporting programs, aimed at the least active population groups. Start to Run, a 6-week training program for novice runners, developed by the Dutch Athletics Organization, is one of these programs. In this study, the effects of Start to Run on health-enhancing physical activity were investigated. Physical activity levels of Start to Run participants were assessed by means of the Short QUestionnaire to ASsess Health-enhancing physical activity (SQUASH) at baseline, immediately after completing the program and six months after baseline. A control group, matched for age and sex, was assessed at baseline and after six months. Compliance with the Dutch physical activity guidelines was the primary outcome measure. Secondary outcome measures were the total time spent in physical activity and the time spent in each physical activity intensity category and domain. Changes in physical activity within groups were tested with paired t-tests and McNemar tests. Changes between groups were examined with multiple linear and logistic regression analyses. In the Start to Run group, the percentage of people who met the Dutch Norm for Health-enhancing Physical Activity, Fit-norm and Combi-norm increased significantly, both in the short- and longer-term. In the control group, no significant changes in physical activity were observed. When comparing results between groups, significantly more Start to Run participants compared with control group participants were meeting the Fit-norm and Combi-norm after six months. The differences in physical activity between groups in favor of the Start to Run group could be explained by an increase in the time spent in vigorous-intensity activities and sports activities. Start to Run positively influences levels of health-enhancing physical activity of participants, both in the short- and longer-term. Based on these results, the use of the organized sports sector as a setting to promote health-enhancing physical activity seems promising.


Estrogen induced prolactin mRNA accumulation in adult male rat pituitary as revealed by in situ hybridization.
Adult male rats were injected with different doses (1, 10, and 100 micrograms) of 17 beta-estradiol daily for 5 days, and the changes in prolactin (PRL) mRNA levels were examined by in situ hybridization and cytoplasmic dot blot hybridization using cloned cDNA for rat prolactin mRNA. An increase in cytoplasmic PRL mRNA content was evident in all the animals treated with estrogen as revealed with cytoplasmic dot blot analysis. There were however, no significant differences in PRL mRNA content among the three estradiol treated groups. Cytoplasmic PRL mRNA was also demonstrated by in situ hybridization on the frozen pituitary sections using a 3H-labeled PRL cDNA probe. The number of grains per cell was increased after estrogen treatment. 3H-thymidine uptake into pituitary cells was also examined in vivo using combined techniques of immunocytochemistry and autoradiography. Although the percentage of immunoreactive PRL cells which took up thymidine in their nuclei increased to more than double after estrogen treatment, the increase in the total number of immunoreactive PRL cells was small. These results suggest that the major effect of estrogen on PRL cells is an increase in the accumulation of PRL mRNA in the individual PRL cells. The number of grains per cell was found to vary from cell to cell, both in control and estrogen treated animals. This variability is discussed in relation to the functional heterogeneity within the PRL cell population.


Betacarotene supplementation protects from photoaging-associated mitochondrial DNA mutation.
Mutations of mitochondrial DNA accumulate during normal aging and can be detected at elevated levels in skin prematurely aged by chronic exposure to ultraviolet (UV) light (photoaging). In normal human fibroblasts, we have previously demonstrated that mtDNA deletions are induced by repetitive exposure to sublethal doses of UVA radiation mediated through singlet oxygen. Betacarotene is a known quencher of ROS and singlet oxygen in particular, and it is widely applied in photoprotective compounds. Therefore we investigated whether in our in vitro system, betacarotene is capable of protecting from the induction of photoaging-associated mtDNA deletions. All-E (trans) betacarotene was tested at doses from 0.25 to 3.0 microM for uptake into cells as well as its protective capacity. Assessment of cellular uptake of all-E betacarotene measured by HPLC revealed a dose dependent increase of intracellular concentrations, as well as an increase in oxidative metabolites. UVA-exposure led to a decrease of all-E-betacarotene, its Z-isomers and oxidative metabolites. Assessment of mtDNA deletions by PCR revealed reduced levels of mtDNA mutagenesis in cells coincubated with betacarotene at concentrations of 0.5 microM and higher. Taken together, these results indicate that betacarotene (i) is taken up into the cell in a dose dependent manner, (ii) interacts with UVA radiation in the cell and (iii) shows protective properties from the induction of a photoaging-associated mtDNA mutation.


Within-person reproducibility and sensitivity to dietary change of C15:0 and C17:0 levels in dried blood spots: Data from the European Food4Me Study.
Previous work highlighted the potential of odd-chain length saturated fatty acids as potential markers of dairy intake. The aim of this study was to assess the reproducibility of these biomarkers and their sensitivity to changes in dairy intake. Fatty acid profiles and dietary intakes from food frequency questionnaires (FFQs) were measured three times over six months in the Food4Me Study. Reproducibility was explored through intra-class correlation coefficients (ICCs) and within-subject coefficients of variation (WCV). Sensitivity to changes in diet was examined using regression analysis. C15:0 blood levels showed high correlation over time (ICC: 0.62, 95% CI: 0.57, 0.68), however, the ICC for C17:0 was much lower (ICC: 0.32, 95% CI: 0.28, 0.46). The WCV for C15:0 was 16.6% and that for C17:0 was 14.6%. There were significant associations between changes in intakes of total dairy, high-fat dairy, cheese and butter and C15:0; and change in intakes of high-fat dairy and cream and C17:0. Results provide evidence of reproducibility of C15:0 levels over time and sensitivity to change in intake of high-fat dairy products with results comparable to the well-established biomarker of fish intake (EPA+DHA).


Inpatient versus other settings for detoxification for opioid dependence.
There are a complex range of variables that can influence the course and subjective severity of opioid withdrawal. There is a growing evidence for the effectiveness of a range of medically-supported detoxification strategies, but little attention has been paid to the influence of the setting in which the process takes place. To evaluate the effectiveness of any inpatient opioid detoxification programme when compared with all other time-limited detoxification programmes on the level of completion of detoxification, the intensity and duration of withdrawal symptoms, the nature and incidence of adverse effects, the level of engagement in further treatment post-detoxification, and the rates of relapse post-detoxification. Electronic databases: the Cochrane Central Register of Controlled Trials (CENTRAL - The Cochrane Library Issue 3, 2004); MEDLINE (January 1966-March 2004); EMBASE (January 1988-March 2004); PsycInfo (January 1967-March 2004); CINAHL (January 1982-March 2004). In addition the Current Contents, Biological Abstracts, Science Citation Index and Social Sciences Index were searched. Randomised controlled clinical trials comparing inpatient opioid detoxification (any drug or psychosocial therapy) with other time-limited detoxification programmes (including residential units that are not staffed 24 hours per day, day-care facilities where the patient is not resident for 24 hours per day, and outpatient or ambulatory programmes, and using any drug or psychosocial therapy). All abstracts were independently inspected by two reviewers (ED & JI) and relevant papers were retrieved and assessed for methodological quality using Cochrane Reviewers' Handbook criteria. Only one study met the inclusion criteria. This did not explicitly report the number of participants in each group that successfully completed the detoxification process, but the published data allowed us to deduce that 7 out of 10 (70%) in the inpatient detoxification group were opioid-free on discharge, compared with 11 out of 30 (37%) in the outpatient group. There was very limited data about the other outcomes of interest. This review demonstrates that there is no good available research to guide the clinician about the outcomes or cost-effectiveness of inpatient or outpatient approaches to opioid detoxification.


Cyclin A and B functions in the early Drosophila embryo.
In eukaryotes, mitotic cyclins localize differently in the cell and regulate different aspects of the cell cycle. We investigated the relationship between subcellular localization of cyclins A and B and their functions in syncytial preblastoderm Drosophila embryos. During early embryonic cycles, cyclin A was always concentrated in the nucleus and present at a low level in the cytoplasm. Cyclin B was predominantly cytoplasmic, and localized within nuclei only during late prophase. Also, cyclin B colocalized with metaphase but not anaphase spindle microtubules. We changed maternal gene doses of cyclins A and B to test their functions in preblastoderm embryos. We observed that increasing doses of cyclin B increased cyclin B-Cdk1 activity, which correlated with shorter microtubules and slower microtubule-dependent nuclear movements. This provides in vivo evidence that cyclin B-Cdk1 regulates microtubule dynamics. In addition, the overall duration of the early nuclear cycles was affected by cyclin A but not cyclin B levels. Taken together, our observations support the hypothesis that cyclin B regulates cytoskeletal changes while cyclin A regulates the nuclear cycles. Varying the relative levels of cyclins A and B uncoupled the cytoskeletal and nuclear events, so we speculate that a balance of cyclins is necessary for proper coordination during these embryonic cycles.


25 years of neuroepidemiology in the Americas.
In the Western Hemisphere, prior to 1950, there were few reliable data on the incidence, geographic distribution or population patterns of diseases of the nervous system. Impressions of prevalence were generally derived from the European literature and from a variety of case reports and relative frequencies based on clinic or hospital admissions or selected autopsy series. Since such data from different sources generally defy comparison, it has been necessary to develop data sources for defined populations to provide incidence and prevalence rates. Such an effort, initially concerned with multiple sclerosis, was launched in 1950 by NIH. International mortality statistics were also collected and analyzed and intensive population surveys in the United States and Canada were planned and carried out. In time, such efforts were extended to cover a wide variety of other chronic and subacute diseases of the nervous system, and support was provided by NIH and voluntary agencies for studies in many other countries. Independently, population studies were developed at various Universities and health care facilities in Latin America. ISSSTE in Mexico City provided a broad experience on multiple sclerosis, ALS, epilepsy, stroke and brain tumor; studies developed in the West Indies on Jamaican Neuropathy; and there were studies in several countries on cysticercosis, Chagas' disease and epilepsy. Also notable are recent studies in Guatemala and plans for geographic comparisons over the broad range of latitude in Chile. The advantages and limitations of mortality statistics will be discussed. The priorities in the selection of the disorders which were to be studied, the procedures which were developed, and the highlights of the results of such investigations will be presented. In addition to descriptive population surveys, case control comparisons and prospective studies are also being developed which seek to identify the risk factors or etiologic determinants that may account for the population pattern which have been observed.


[Distribution of reticulospinal fibers and theri endings in the lumbar segments of the rat spinal cord].
After stereotaxis lexions in the nucleus reticularis gigantocellularis of the modulla oblongata and nucleus reticularis pontis caudalis, the distribution of degenerating nerve fibers in the lumbar segments of the spinal cord has been studied by silver impregnation methods of Nauta and Fink-Heimer. Degenerating reticulo-spinal fibers and fragments of axonal terminations were found in the area of n. motorius ventro-medialis and n. motorius ventro-lateralis, as well as partly in n. motorius dorso-lateralis close to motoneurons and their dendrites. Mainly they pass into layers VII and VIII. This fact indicates the existence of direct-reticulo-motoneuronal synaptic connections in rats, which coincides with electrophysiological data.


Immunogenicity and safety of autoclaved Leishmania major plus BCG vaccine in healthy Sudanese volunteers.
In a longitudinal study in the epidemiology of Leishmania donovani infection in an endemic focus in eastern Sudan, we observed that previous exposure or infection with Leishmania major appeared to protect against visceral leishmaniasis caused by L. donovani. We therefore conducted a study to test the safety and immunogenicity of a vaccine consisting of autoclaved L. major (ALM) plus BCG in inducing protection in vaccinated individuals. Leishmanin-negative healthy Sudanese volunteers were enrolled in the study and were divided into three groups: group (A) received ALM+BCG, group (B) received BCG alone, and group (C) received the vaccine diluent. The subjects were examined for their clinical and immunological responses before intervention, following intervention and 6-8 weeks after vaccination. Vaccinated subjects (group A) developed localized reactions at the sites of vaccine inoculation that ulcerated and healed within 4-6 weeks; 61.6% of them converted to leishmanin reactive following vaccination. Only one subject in group (C) became leishmanin-positive. A total 76.9% of the vaccinated volunteers in group (A) produced significant levels of interferon-gamma in response to L. major antigen. The vaccine produced significant cellular immune responses that may protect against natural challenge. None of the groups had systemic reactions and all the reactions observed in the vaccinated group were comparable with the BCG-vaccinated group.


Dynamics of support perceptions among couples coping with cardiac illness: the effect on recovery outcomes.
The current prospective study explored how male cardiac patients' perceptions of received support (i.e., active engagement, protective buffering, and overprotection) moderated the associations between female partners' perceptions of provided support and patients' recovery outcomes: psychological well-being, cholesterol levels, and smoking cessation. Couples (N = 86) completed surveys at the initial hospitalization after patients' Acute Coronary Syndrome (ACS), and 1 and 6 months later. Partners' ways of providing support and patients' concurrent perceptions of these ways were measured using the Ways of Giving Support Questionnaire; patients' depressive and anxiety symptoms were measured using the Brief Symptom Inventory (BSI). Patients' cholesterol levels were assessed during hospitalization and 6 months later, and smoking habits were reported by the patients. Female partners' protective buffering was positively associated with male patients' depressive symptoms at follow-up only when male patients' own perceptions of partners' protective buffering were low. Female partners' active engagement was positively associated with better odds for male patients' cessation of smoking only when patients' own perceptions of partners' active engagement were high. Finally, female partners' overprotection was associated with higher levels of male patients' harmful blood lipids at follow-up, but only when patients' own perceptions of partners' overprotection were high. As hypothesized, the effect of partners' perceptions of support provided on patients' recovery was moderated by patients' own perceptions of the support received. The effect of this interaction was determined by the specific types of support provided or received and by the specific recovery outcome that was measured. The clinical and theoretical implications of the findings are discussed.


Living with late deafness: insight from between worlds.
The psychosocial impact of 'late' deafness in adults has received little research attention. The aim of this study was to examine the views of people with experience of late deafness living in the UK. Eight participants (six male; age range 33 to 60) were interviewed by a researcher who had undergone appropriate communication skills training. In-depth, face-to-face, semi-structured interviews were conducted and transcribed. Data were analysed using framework analysis to identify themes. Becoming deafened was a devastating experience for many participants who experienced severe psychological, social, and employment consequences. They no longer felt they belonged in the hearing world, and neither did they belong in the prelingually deaf world. Onset of deafness had left them in a twilight zone between worlds and had robbed them of their identity. Whilst the support received from health and social care professionals was mixed, all participants valued the peer support and training received on an intensive rehabilitation programme delivered by deafened people. Findings provide insight into deafened peoples' psychosocial experiences and emphasize the need for support and advice.


Cardiac resynchronization therapy: refocus on the electrical substrate.
Cardiac resynchronization therapy (CRT) is an established treatment for selected heart failure patients with conduction disease. Many studies aimed at quantifying mechanical dyssynchrony in CRT candidates when it became apparent that 30-50% of CRT recipients showed no improvement after implantation. As these, often echocardiography-based, measurements have not yet succeeded in estimating the mechanical substrate in an accurate and reproducible manner, interest in electrical substrate has renewed. In this review, current knowledge concerning electrical substrate in CRT candidates will be explored and applied to current CRT practice, highlighting why the electrical substrate is both essential and sufficient for successful CRT. Finally, novel ways to better measure and treat the electrical substrate are discussed.


Barriers and facilitators to the uptake of computerized clinical decision support systems in specialty hospitals: protocol for a qualitative cross-sectional study.
Computerized clinical decision support systems (CDSSs) have been shown to improve the efficiency and quality of patient care by connecting healthcare professionals with high quality, evidence-based information at the point-of-care. The mere provision of CDSSs, however, does not guarantee their uptake. Rather, individual and institutional perceptions can foster or inhibit the integration of CDSSs into routine clinical workflow. Current studies exploring health professionals' perceptions of CDSSs focus primarily on technical and usability issues, overlooking the social or cultural variables as well as broader administrative or organizational roles that may influence CDSS adoption. Moreover, there is a lack of data on the evolution of perceived barriers or facilitators to CDSS uptake across different stages of implementation. We will conduct a qualitative, cross-sectional study in three Italian specialty hospitals involving frontline physicians, nurses, information technology staff, and members of the hospital board of directors. We will use semi-structured interviews following the Grounded Theory framework, progressively recruiting participants until no new information is gained from the interviews. CDSSs are likely to become an integral and diffuse part of clinical practice. Various factors must be considered when planning their introduction in healthcare settings. The findings of this study will guide the development of strategies to facilitate the successful integration of CDSSs into the regular clinical workflow. The evaluation of diverse health professionals across multiple hospital settings in different stages of CDSS uptake will better capture the complexity of roles and contextual factors affecting CDSS uptake.


Preparation of protein samples for NMR structure, function, and small-molecule screening studies.
In this chapter, we concentrate on the production of high-quality protein samples for nuclear magnetic resonance (NMR) studies. In particular, we provide an in-depth description of recent advances in the production of NMR samples and their synergistic use with recent advancements in NMR hardware. We describe the protein production platform of the Northeast Structural Genomics Consortium and outline our high-throughput strategies for producing high-quality protein samples for NMR studies. Our strategy is based on the cloning, expression, and purification of 6×-His-tagged proteins using T7-based Escherichia coli systems and isotope enrichment in minimal media. We describe 96-well ligation-independent cloning and analytical expression systems, parallel preparative scale fermentation, and high-throughput purification protocols. The 6×-His affinity tag allows for a similar two-step purification procedure implemented in a parallel high-throughput fashion that routinely results in purity levels sufficient for NMR studies (>97% homogeneity). Using this platform, the protein open reading frames of over 17,500 different targeted proteins (or domains) have been cloned as over 28,000 constructs. Nearly 5000 of these proteins have been purified to homogeneity in tens of milligram quantities (see Summary Statistics, http://nesg.org/statistics.html), resulting in more than 950 new protein structures, including more than 400 NMR structures, deposited in the Protein Data Bank. The Northeast Structural Genomics Consortium pipeline has been effective in producing protein samples of both prokaryotic and eukaryotic origin. Although this chapter describes our entire pipeline for producing isotope-enriched protein samples, it focuses on the major updates introduced during the last 5 years (Phase 2 of the National Institute of General Medical Sciences Protein Structure Initiative). Our advanced automated and/or parallel cloning, expression, purification, and biophysical screening technologies are suitable for implementation in a large individual laboratory or by a small group of collaborating investigators for structural biology, functional proteomics, ligand screening, and structural genomics research.


Lysine and Arginine Protein Post-translational Modifications by Enhanced DIA Libraries: Quantification in Murine Liver Disease.
Proteoforms containing post-translational modifications (PTMs) represent a degree of functional diversity only harnessed through analytically precise simultaneous quantification of multiple PTMs. Here we present a method to accurately differentiate an unmodified peptide from its PTM-containing counterpart through data-independent acquisition-mass spectrometry, leveraging small precursor mass windows to physically separate modified peptidoforms from each other during MS2 acquisition. We utilize a lysine and arginine PTM-enriched peptide assay library and site localization algorithm to simultaneously localize and quantify seven PTMs including mono-, di-, and trimethylation, acetylation, and succinylation in addition to total protein quantification in a single MS run without the need to enrich experimental samples. To evaluate biological relevance, this method was applied to liver lysate from differentially methylated nonalcoholic steatohepatitis (NASH) mouse models. We report that altered methylation and acetylation together with total protein changes drive the novel hypothesis of a regulatory function of PTMs in protein synthesis and mRNA stability in NASH.


Insulin receptor substrate-1 regulates the transformed phenotype of BT-20 human mammary cancer cells.
Although originating from a human breast cancer, BT-20 cells do not form colonies in soft agar. BT-20 cells do not express insulin receptor substrate-1 (IRS-1), which is known to promote both normal and abnormal growth and to inhibit differentiation. Stable expression of IRS-1 confers to BT-20 cells the ability to form colonies in soft agar. BT-20 cells form tumors in xenografts in mice, but the size of tumors is twice as large when the cells express IRS-1. The increased transformed phenotype is characterized by occupancy of the rDNA and cyclin D1 promoters by IRS-1 and the activation of the cyclin D1, c-myc, and rDNA promoters. In addition, the retinoblastoma protein, which is detectable in the rDNA promoter of quiescent BT-20/IRS-1 cells, is replaced by IRS-1 after insulin-like growth factor-I stimulation. Our results indicate that in BT-20 human mammary cancer cells, expression of IRS-1 activates promoters involved in cell growth and cell proliferation, resulting in a more transformed phenotype. Targeting of IRS-1 could be effective in inhibiting the proliferation of mammary cancer cells.


Regulation of Glycosylphosphatidylinositol-Anchored Protein (GPI-AP) Expression by F-Box/LRR-Repeat (FBXL) Protein in Wheat (Triticum aestivum L.).
F-box proteins are substrate recognition components of the Skp1-Cullin-F-box (SCF) complex, which performs many important biological functions including the degradation of numerous proteins via the ubiquitin-26S proteasome system. In this study, we isolated the gene encoding the F-box/LRR-repeat (FBXL) protein from wheat (Triticum aestivum L.) seedlings and validated that the TaFBXL protein is a component of the SCF complex. Yeast two-hybrid assays revealed that TaFBXL interacts with the wheat glycosylphosphatidylinositol-anchored protein (TaGPI-AP). The green fluorescent protein (GFP) fusion protein of TaFBXL was detected in the nucleus and plasma membrane, whereas that of TaGPI-AP was observed in the cytosol and probably also plasma membrane. yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays revealed that TaFBXL specifically interacts with TaGPI-AP in the nucleus and plasma membrane, and TaGPI-AP is targeted by TaFBXL for degradation via the 26S proteasome system. In addition, TaFBXL and TaGPI-AP showed antagonistic expression patterns upon treatment with indole-3-acetic acid (IAA), and the level of TaGPI-AP was higher in tobacco leaves treated with both MG132 (proteasome inhibitor) and IAA than in leaves treated with either MG132 or IAA. Taken together, our data suggest that TaFBXL regulates the TaGPI-AP protein level in response to exogenous auxin application.


Comparative physiologic performance of polymethyl methacrylate and gas-permeable contact lenses.
A short-term study of the physiologic response of the cornea to polymethyl methacrylate (PMMA) contact lenses and hard, gas-permeable contact lenses of different materials (polycon and Menicon 02), thicknesses, and fittings, was carried out on nine unadapted subjects using one eye as the experimental eye and the other as the control eye (no lens). Changes in corneal thickness were monitored at seven locations across the cornea using an electronic micropachymeter. It was found that gas-permeable lenses produced less corneal swelling than the PMMA lenses. However, lens thickness and the fit of the lens were still important control variables with these gas-permeable lenses. The thin gas-permeable lens produced less swelling than thicker lenses, and the flatter fitting gas-permeable lenses produced less swelling than the steeper fitting lenses. The swelling with all the hard lenses studied was confined primarily to the central 6 mm of the cornea.


Prostacyclin infusion in patients with acute myocardial infarction.
To investigate whether prostacyclin protects ischaemic myocardium in humans the effect of prostacyclin or placebo was studied in two groups of patients with acute myocardial infarction who presented within six and 16 hours of the onset of symptoms. Intravenous infusion of prostacyclin or placebo was started immediately after admission at a rate corresponding to 4-5 ng/kg/minute. The infusion was maintained for 72 hours. Clinical status, electrocardiograms, plasma enzyme activity, infarct extension during the infusion, and reinfarction after the infusion were studied. Prostacyclin was well tolerated by most patients: neither systemic blood pressure nor heart rate differed between the two groups. In the 11 patients who received treatment within six hours of the onset of symptoms prostacyclin significantly lowered the maximum plasma activities of creatine kinase MB and lactate dehydrogenase. In the 19 patients who received treatment 6-16 hours after the onset of symptoms prostacyclin had no such effect. None of the patients receiving prostacyclin had an extension of the infarction during the infusion, whereas four patients receiving placebo did; this difference was significant. These data are the first to provide evidence that prostacyclin might limit myocardial injury in patients with acute myocardial infarction.


Freeze-drying of spermatozoa.
Bull semen was diluted to a concentration of 2 X 10(8) cells/ml, cooled to 5 degrees C in 5 h, frozen in 0.025 ml spheres on the surface of solid carbon dioxide, and stored in liquid nitrogen. 50% by volume of the diluent was 325 m0sm Tes:N-tris (hydroxymethyl) methyl-2-amino ethane sulfonic acid titrated to pH 7.2 with 325 m0sm Tris:tris (hydroxymethyl) amino methane. The diluent also contained 30% by volume isotonic sodium citrate and 20% by volume egg yolk. The frozen spermatozoa were freeze-dried in 400 mg quantities in test tubes at -50 degrees C with a condenser at -196 degrees C. Moisture content was determined by weighing the individual samples before and after freeze-drying. Drying the samples for several days at 20 degrees C removed 868 mg from each gram of frozen material and this was considered the zero moisture level. Samples were stored at 20 degrees C or -196 degrees C after freeze-drying. The freezer-dried samples were rehydrated by flooding with five times their original volume of isotonic sodium citrate. Tests of the recovered spermatozoa included percentage motile cells, acrosome damage, enzyme release, protein denaturation, hypotonic swelling and fertility testing. Sperm motility decreased with dryness until it reached zero at 3% moisture. Acrosome morphology and enzyme release appeared normal down to 1% moisture. Preliminary results showed some fertility at all levels of dryness with -196 degrees C storage, and fertility at less than 1% moisture with 20 degrees C storage. Additional fertility testing is underway.


What Time Alone Offers: Narratives of Solitude From Adolescence to Older Adulthood.
Solitude - the state of being alone and not physically with another - can be rewarding. The present research explored the potential benefits of solitude from a pragmatist approach: a ground-up, top-down perspective that is receptive to new knowledge but informed by theory. Participant recruitment was stratified by age and gender, and the sample involved 2,035 individuals including adolescents (13-16 years), adults (35-55 years), or older adults (65+ years). Data were analyzed with a mixed-methods approach. Coded themes from brief narratives about solitude were extracted, and their frequencies (i.e., their salience to participants) were compared across the lifespan. Themes were then correlated with two indicators of well-being in solitude: self-determined motivation for solitude and peaceful mood. Several prominent themes emerged when talking about time spent in solitude. With the exception of feeling competent in solitude, which was described frequently but consistently unrelated to self-reported well-being regardless of age, benefits of solitude tended to shift over the lifespan. Some qualities, such as a sense of autonomy (self-connection and reliance; absence of pressure), were salient and consequential for everyone, but increasingly so from adolescence to older adulthood. Older adults also reported feeling most peaceful in solitude and described their social connection and alienation less frequently, suggesting they see solitude and social time as more distinct states. Findings are discussed in light of existing work on solitude across the lifespan, and theoretical frameworks that spoke well to the data (e.g., self-determination theory).


New insights on the neuroendocrine control of puberty and seasonal breeding in female sheep.
Timing of puberty has a great influence on animal productivity. For example, reproduction in sheep can be affected by seasonality, leading to fluctuations in availability of animal products. Therefore, optimization of birth dates would improve reproductive success in sheep. Since the discovery of the major role of kisspeptin and Kiss1R, its cognate receptor, in reproductive function, there are new opportunities for interventions. Repeated or continuous administration of native kisspeptin are able to hasten puberty and induce ovulation during breeding and non-breeding seasons of sheep. However, due to the short half-life of kisspeptin, protocols involving native kisspeptin are usually proof of concept, but not practical under field conditions. Consequently, there are efforts to develop kisspeptin analogues capable of replicating effects of repeated/continuous administration of native kisspeptin. In this review, we intended to provide a comprehensive summary of the neuroendocrine requirements for puberty onset and ovulation in adult ewes, focusing on kisspeptin, its physiological effects and responses to its analogues on reproductive function in ewes.


Synthetic ligaments. Current status.
Many techniques for ligamentous reconstruction have been developed in recent years. In the United States, injuries of the knee have been increasingly treated with innovative methods of surgical reconstruction, most of which have used normal structures. There are obvious theoretic advantages in using synthetic materials that might simplify surgery, spare normal tissues, and possibly facilitate stronger repairs. To these ends, several synthetic substances have been used experimentally and clinically. This is a brief summary of eight of the materials that have been or are being investigated in the United States. Some are no longer in use, others are currently being used in clinical trials. As of this writing, only the Gortex ligament has received a general device release from the Food and Drug Administration (FDA).


Probing the Effect of Cuttings Particle Size on the Friction and Wear Mechanism at the Casing Friction Interface: A Molecular Dynamics Study.
Cuttings particles of different sizes in the drilling fluid are the leading cause of wear at the casing and drill pipe joints, and diamond-like carbon (DLC) films have excellent research potential in reducing tool wear due to their ultra-low friction coefficient and high wear resistance. In this paper, a corresponding molecular dynamics model was developed using LAMMPS to investigate the effect of silica particles of different particle sizes on the friction and wear mechanisms of Fe/DLC friction pairs at the microscale. The results show that small cuttings particles in a dry environment are more likely to cause interface wear between the casing and drill pipe joint, while in a water environment, the opposite is true. The main reason is that small particles in a dry environment have smaller contact areas and greater indentation depth, leading to greater wear at the friction interface. The movement of water molecules in the water environment will promote the composite movement of large particles, thereby exacerbating the wear of the interface. Moreover, the relevant research results at the micro-scale indicate that DLC films can effectively reduce wear, which provides theoretical support for its application in drill pipe joints.


Animal Cap Isolation from Xenopus laevis.
INTRODUCTIONThis protocol presents a method for isolating Xenopus laevis animal cap cells, which are cells situated around the animal (pigmented) pole of a blastula or very early gastrula-stage embryo. This tissue is fated to become cement gland/neurectoderm on the dorsal side of the embryo and epidermis on the ventral side. Animal caps are composed of pluripotent cells that can be induced to form endodermal, mesodermal, or ectodermal cell types, and can therefore serve as a useful substrate to assess the activity of various inducing factors. Caps from embryos that have been injected with expression constructs can also be removed and analyzed to assess the activity of various genes. In addition, caps can be used in conjugation (induction) assays.


Causes of reduced clutch size in a tidal marsh endemic.
We tested three hypotheses of clutch size variation in two subspecies of the swamp sparrow (Melospiza georgiana georgiana and M. g. nigrescens). Swamp sparrows follow the pattern of other estuarine endemics, where clutch size is smaller among tidal salt marsh populations (M. g. nigrescens) than their closest inland relatives (M. g. georgiana). Our results support predation risk and temperature, but not adult survival, as explanations of this pattern in swamp sparrows. Coastal nests were twice as likely to fail as inland nests, and parental activity around the nest site was positively related to clutch size at both sites. When brood size was controlled for, coastal adults visited nests less often and females vocalized less frequently during visits than inland birds, which may decrease nest detectability to predators. Coastal parents waited longer than inland birds to feed offspring in the presence of a model nest predator, but there was no difference in their response to models of predators of adults, as would be expected if coastal birds possessed increased longevity. Additionally, coastal females laid more eggs than inland females over a single season, following a within-season bet-hedging strategy rather than reducing within-season investment. Coastal territories experienced ambient air temperatures above the physiological zero of egg development more often, and higher temperatures during laying correlated with smaller clutches and increased egg inviability among coastal birds. Similar effects were not seen among inland nests, where laying temperatures were generally below physiological zero. Both subspecies showed an increase in hatching asynchrony and a decrease in apparent incubation length under high temperatures. Coastal individuals, however, showed less hatching asynchrony overall despite higher temperatures. Both air temperatures during laying and predation risk could potentially explain reduced clutch size in not only coastal plain swamp sparrows, but also other tidal marsh endemics.


Retention of intratumor injections of cisplatinum in murine tumors and the impact on laser thermal therapy for cancer treatment.
Recent studies using murine models of human squamous cell carcinoma (SCCA) have revealed a significant improvement in survival and cure rate of animals transplanted with human SCCA when treated with a combination of intratumor injections of chemotherapy and laser induced thermal therapy (LITT). These preliminary results suggest that this novel combination therapy may lead to improved clinical response compared to either treatment modality alone. Using a murine model of human SCCA we investigated two different modes of intratumor injection of cisplatin: a sustained-release cisplatin gel implant (CDDP/gel) versus cisplatin in solution (CDDP) at varying doses (range 1-3 mg/ml). In addition, we tested CDDP/gel combined with LITT. Results showed optimal drug concentration (30-300 nM) at tumor margins up to 4 h after injection of CDDP/gel implant compared to 3 nM at 5 min after injection with CDDP solution. Combined CDDP/gel and laser therapy significantly decreased tumor volume (P<0.05), with recurrence in only 25% of animals tested, compared to 78% tumor regrowth after LITT alone. These results suggest that laser chemotherapy may be an effective treatment for head and neck SCCA.


Individualizing justice after Atkins.
On August 6, 2005, newspapers and other media outlets reported that Daryl Atkins had been determined by a Virginia jury not to be retarded and therefore was mentally competent to receive the death penalty. A judge immediately scheduled his execution for December. Atkins, of course, is the convicted murderer whose case three years earlier had led the U.S. Supreme Court, in a landmark ruling, to declare that mentally retarded offenders are constitutionally exempt from the death penalty. While a bitter irony for Atkins, his family, and supporters, the Virginia jury's finding suggests that the practical effects of the Supreme Court's decision are less dramatic than many had anticipated. It shows that mere labels need not be determinative and that judges and juries as well as mental health experts called to assist them in capital cases can continue to work toward an individualized brand of justice.


Yeast secretory mutants that block the formation of active cell surface enzymes.
Yeast cells secrete a variety of glycosylated proteins. At least two of these proteins, invertase and acid phosphatase, fail to be secreted in a new class of mutants that are temperature-sensitive for growth. Unlike the yeast secretory mutants previously described (class A sec mutants; Novick, P., C. Field, and R. Schekman, 1980, Cell., 21:205-420), class B sec mutants (sec 53, sec 59) fail to produce active secretory enzymes at the restrictive temperature (37 degrees C). sec 53 and sec 59 appear to be defective in reactions associated with the endoplasmic reticulum. Although protein synthesis continues at a nearly normal rate for 2 h at 37 degrees C, incorporation of [3H]mannose into glycoprotein is reduced. Immunoreactive polypeptide forms of invertase accumulate within the cell which have mobilities on SDS PAGE consistent with incomplete glycosylation: sec 53 produces little or no glycosylated invertase, and sec 59 accumulates forms containing 0-3 of the 9-10 N-linked oligosaccharide chains that are normally added to the protein. In addition to secreted enzymes, maturation of the vacuolar glycoprotein carboxypeptidase Y, incorporation of the plasma membrane sulfate permease activity, and secretion of the major cell wall proteins are blocked at 37 degrees C.


Modulation of agrin-induced acetylcholine receptor clustering by extracellular signal-regulated kinases 1 and 2 in cultured myotubes.
Agrin released by motoneurons induces and/or maintains acetylcholine receptor (AChR) clustering and other aspects of postsynaptic differentiation at the vertebrate neuromuscular junction. Agrin acts by binding and activating a receptor complex containing LDL receptor protein 4 (Lrp4) and muscle-specific kinase (MuSK). Two critical downstream components of this signaling cascade, Dox-7 and rapsyn, have been identified. However, additional intracellular essential elements remain unknown. Prior observations by others and us suggested antagonistic interactions between agrin and neuregulin-1 (Nrg-1) signaling in cultured myotubes and developing muscle fibers in vivo. A hallmark of Nrg-1 signaling in skeletal muscle cells is the activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2). ERK1/2 are also activated in most cells by phorbol 12-myristate 13-acetate, a classical inhibitor of agrin-induced AChR clustering in myotubes. Here, it was investigated whether agrin activates ERK1/2 directly and whether such activation modulates agrin-induced AChR clustering. Agrin induced a rapid but transient activation of ERK1/2 in myotubes that was Lrp4/MuSK-dependent. However, blocking this ERK1/2 activation did not prevent but potentiated AChR clustering induced by agrin. ERK1/2 activation was dispensable for Nrg-1-mediated inhibition of the AChR clustering activity of agrin, but was indispensable for such activity by phorbol 12-myristate 13-acetate. Together, these results suggest agrin-induced activation of ERK1/2 is a negative modulator of agrin signaling in skeletal muscle cells.


[Genetic polymorphisms and genetic effects of IGF2 gene in pigs].
Single nucleotide polymorphisms of the exon7, 8, 9 of the porcine IGF2 gene were tested using PCR-SSCP. Genetic effects of the IGF2 gene on birth weight, weaning weight, six-month weight and six-month backfat thickness were analyzed. On the basis of the published DNA sequence (AY044828) of the porcine IGF2 gene, three pairs of primers were designed, and one polymorphism, three genotypes were found in the PCR product amplified with Ex8 primers. C-->T transition at 53 site of exon8 was detected by sequencing the homozygotes. The results showed that: firstly, the genotype distribution was not consistent among the different pig breeds. No significant differences (P > 0.05) were found in the genotype distribution when compared the Landrace with the Large White , the Laiwu with the Dapuliang, and the Yimeng with the Licha Black pig breeds, whereas significant differences (P lt; 0.01) were found among other tested pig breeds in the genotype distribution. Secondly, on the basis of the fixed effect model, significant differences (P lt; 0.05) were found in birth weight and six-month backfat thickness, whereas no significant differences (P > 0.05) existed in weaning weight and six-month weight. Thirdly, using least square analysis, it was shown that individuals of the BB genotype have significantly lower (P lt; 0.05) birth weight than those of AA and AB genotypes, with the order being AB > AA > BB ; Individuals of the AA genotype have significantly lower (P lt; 0.05) six-month backfat thickness than those of AB and BB genotypes, with the order being BB > AB > AA. These results suggest that IGF2 gene has significant effects on individual birth weight and backfat thickness. The IGF2 gene can be used in marker-assisted selection to accelerate the breeding progress.


Atypical central pain processing in sensory modulation disorder: absence of temporal summation and higher after-sensation.
Sensory over-responsivity (SOR), a subtype of the proposed sensory modulation disorder (SMD), is characterized by over-responsiveness to stimuli in several sensory modalities. SMD individuals demonstrate abnormal responses to naturally occurring stimuli in a manner that interferes with daily life participation. Previous psychophysical testing of the somatosensory system revealed that SOR individuals rated pain sensations higher than controls, demonstrating hyperalgesia that can be centrally mediated. Temporal summation (TS) of second pain and after-sensation are manifestations of central sensitization; therefore, this study explored these measures for better characterization of central pain processing in SOR. Twelve SOR adults and 12 healthy controls participated. TS was produced by a train of fifteen repetitive heat pulses, 0.7 s duration each, and 2 s of inter-stimulus interval, applied to the thenar-eminence, while four pain ratings were obtained. An after-sensation was then measured for 5 min, obtaining six pain ratings. No TS of pain was indicated in the SOR group (SOR: p = 0.36; control: p < 0.001). Further, while controls reported a gradual disappearance of pain after-sensation, individuals with SOR continued to report pain for the duration of the 5 min measured (p = 0.002). These results demonstrate an atypical response pattern, suggesting alteration in pain processing and/or modulation at a central level in individuals with SOR. These possible neural changes may manifest themselves as interference with daily functioning as well as shed light on some of the between-subject variability seen in psychophysical testing in non-painful subjects.


Correlation between pretreatment viral sequences and the emergence of lamivudine resistance in hepatitis B virus infection.
The emergence of amino acid or nucleotide substitutions leads to lamivudine resistance in hepatitis B virus (HBV) infected patients. The aim of this study was to investigate whether viral sequences help predict the emergence of lamivudine resistance. The study subjects comprised 59 consecutive patients infected with HBV treated with daily therapy of 100 mg lamivudine. Among those, 32 patients with adequate pretreatment serum preservation were investigated for the correlation between viral amino acid substitutions and the appearance of lamivudine resistance with consideration of clinical background by determining dominant HBV full open reading frames. Viral resistance to lamivudine emerged in 28 of 59 patients (47%) in a median period of 2.45 years. Sequence comparisons of HBV genomes between patients who later developed lamivudine resistance and patients who did not revealed the existence of significant differences between the two groups in the pre-S1 84 (P = 0.042), pre-S2 1 (P = 0.017) and 22 (P = 0.015), and polymerase tp 95 (P = 0.046), judged by a log-rank test. Viral sequence analyses revealed the presence of amino acid substitutions in HBV pre-S1 and pre-S2 that may be associated with the emergence of lamivudine resistance during chronic HBV infection.


Space charge neutralization by electron-transparent suspended graphene.
Graphene possesses many fascinating properties originating from the manifold potential for interactions at electronic, atomic, or molecular levels. Here we report measurement of electron transparency and hole charge induction response of a suspended graphene anode on top of a void channel formed in a SiO2/Si substrate. A two-dimensional (2D) electron gas induced at the oxide interface emits into air and makes a ballistic transport toward the suspended graphene. A small fraction (>~0.1%) of impinging electrons are captured at the edge of 2D hole system in graphene, demonstrating good transparency to very low energy (<3 eV) electrons. The hole charges induced in the suspended graphene anode have the effect of neutralizing the electron space charge in the void channel. This charge compensation dramatically enhances 2D electron gas emission at cathode to the level far surpassing the Child-Langmuir's space-charge-limited emission.


Production of recombinant HIV-1/HBV virus-like particles in Nicotiana tabacum and Arabidopsis thaliana plants for a bivalent plant-based vaccine.
Human immunodeficiency virus (HIV-1) and hepatitis B virus (HBV) spread via similar transmission pathways, and infection by HBV occurs in up to 32% of HIV-1 cases. Here, we describe the successful expression of novel recombinant HIV-1/HBV virus-like particles (VLPs) in Nicotiana tabacum and Arabidopsis thaliana. The production levels and quality of the recombinant VLPs were comparable in the two plants, showing that parameters intrinsic to the recombinant proteins determined their assembly into VLPs. These heterologous VLPs can be used in a bivalent anti-HIV-1/-HBV vaccine, administrated via ingestion of transgenic plants.


Physiological role of the interaction between CARMIL1 and capping protein.
The regulation of free barbed ends is central to the control of dynamic actin assembly and actin-based motility in cells. Capping protein (CP) is known to regulate barbed ends and control actin assembly in cells. The CARMIL family of proteins can bind and inhibit CP in vitro, but the physiological significance of the interaction of CARMIL with CP in cells is poorly understood. Mammalian cells lacking CARMIL1 have defects in lamellipodia, macropinocytosis, cell migration, and Rac1 activation. Here we investigate the physiological significance of the CARMIL1-CP interaction, using a point mutant with a well-defined biochemical defect. We find that the CARMIL1-CP interaction is essential for the assembly of lamellipodia, the formation of ruffles, and the process of macropinocytosis. In contrast, the interaction of CARMIL1 with CP shows little to no importance for other functions of CARMIL1, including localization of CARMIL1 to the membrane, activation of Rac1, and cell migration. One implication is that lamellipodia are only marginally important for cell migration in a wound-healing model. The results also suggest that the ability of CARMIL1 to inhibit CP in cells may be regulated.


Deep vein thrombosis.
This article reviews the incidence, causes, risk factors, diagnosis, treatment, complications, and prevention of deep vein thrombosis (DVT). Various employees, including long-distance drivers or travelers, sedentary office workers, females taking estrogen, those who are obese, those who have cancer, and those who are hospitalized, may be at risk for developing this condition. Genetic testing can determine some inherited factors predisposing workers to clotting. Low-molecular weight heparins can be used to manage DVT on an outpatient basis.


Cardiac myxoma: A surgical experience of 38 patients over 9 years, at SSKM hospital Kolkata, India.
Cardiac myxoma is the most common benign intracardiac tumor. We studied its clinical presentation, morbidity, mortality and recurrence following surgery over a period of 9 years. This study was performed at cardiothoracic and vascular surgery department of a tertiary level hospital of eastern India, Seth Sukhlal Karnani Memorial hospital, Institute of Post Graduate Medical Education and Research Kolkata. Near 6000 cardiac cases were operated at our center over this period. Preoperative diagnosis was made with clinical presentation and preoperative echocardiography. Complete tumor excision was done and all patients were followed up for recurrence and complications. A total of 38 cases of cardiac myxoma were operated over a period from October 2002 to October 2011. Cardiac myxoma constituted about 0.6% of all cardiac cases operated at our institute. This most commonly presented at fifth decade of life. Of these, 35 cases were left atrial and 2 cases were right atrial, and 1 case was having both atrial involvements. The left atrial myxoma mostly presented as mitral stenosis and very few presented with embolic and constitutional symptoms. No death or recurrence was observed during the follow up period. Cardiac myxomas form a very small percentage of the cardiac cases. A high index of suspicion is essential for diagnosis. Echocardiography is the ideal diagnostic tool as also for follow-up. Immediate surgical treatment is indicated in all patients. Cardiac myxomas can be excised with a low rate of mortality and morbidity.


Understanding the illness representations of young people with anxiety and depression: A qualitative study.
Many young people with anxiety or depression drop out of treatment early, and/or leave treatment without showing measurably improved symptom levels. To enhance treatment engagement and effectiveness, it is critical to better understand how young people's perceptions of the symptoms, causes, consequences, treatability, and course of their anxiety and depression influence engagement. This study aimed to provide a qualitative account of illness perceptions among youth with anxiety and depression by applying the Common Sense Model of Self-Regulation (CSM), which was developed in physical health contexts. Semi-structured interviews were conducted with 26 young people (aged 16-24, 73% female) with a history of anxiety and/or depression. Interviews were analysed using a combination of theory- and data-driven analysis techniques, consisting primarily of deductive thematic analysis. The five themes broadly mapped onto the dimensions of the CSM, suggesting parallels in how mental and physical health problems are perceived. Anxiety and depression were viewed as non-linear, relapsing and remitting, but lifelong conditions, with a fluctuating and complex path to recovery and coping. Youth described pervasive negative impacts on their lives, but also described some positive aspects. Better understanding of young people's illness beliefs has the potential to open a range of intervention possibilities by prioritizing young people's illness perceptions over the clinician's understanding and the supposed objective condition severity and trajectory. Although this study supported a common structure of illness beliefs, the content of these beliefs was idiosyncratic and specific to anxiety and depression, suggesting the need to develop a valid tool to measure illness perceptions in this group. Our findings suggest that illness perceptions are complex, highly idiosyncratic, and specific to youth anxiety and depression. Given the complexity of these beliefs and the known association with important treatment- and health-related outcomes, it is important that clinical formulation incorporates young people's illness belief models, including their perceptions of symptoms, cause, timeline to recovery, consequences, and personal and treatment control. To increase help-seeking, treatment engagement and adaptive coping strategies, therapy should work to a shared understanding of illness beliefs. Increasing congruence between the belief models of young people, families, and clinicians may serve to improve treatment benefits and address the unmet mental health needs of young people.


Phage display of cDNA libraries: enrichment of cDNA expression using open reading frame selection.
Phage display technologies are powerful tools for selecting binding ligands against purified molecular targets, live cells, and organ vasculature. However, the selection of natural ligands using phage display has been limited because of significant problems associated with the display of complex cDNA repertoires. Here we describe the use of cDNA fragmentation and open reading frame (ORF) selection to display a human placental cDNA library on the pIII coat protein of filamentous phage. The library was enriched for ORFs by selecting cDNA-beta-lactamase fusion proteins on ampicillin, resulting in a cDNA population having 97% ORFs. The ORF-selected cDNAs were fused to pIII in the phagemid vector, pUCMG4CT-198, and the library was rescued with a pIII-deleted helper phage for multivalent display. The resulting phagemid particle library consisted of 87% ORFs, compared to only 6% ORFs when prepared without ORF selection. Western blot analysis indicated cDNA-pIII fusion protein expression in eight out of nine ORF clones tested, and seven of the ORF encoded peptides were displayed multivalently. The high level of cDNA expression obtained by ORF selection suggests that ORF-enriched phage cDNA libraries prepared by these methods will be useful as functional genomics tools for identifying natural ligands from various source tissues.


Specificity determinants of CYP1B1 estradiol hydroxylation.
Cytochrome P450 (P450)-catalyzed oxidation of the aromatic ring of estradiol can result in 2- or 4-hydroxylation. Which of these products is formed is biologically important, as the 4-hydroxylated metabolite is carcinogenic, whereas the 2-hydroxylated metabolite is not. Most human P450 enzymes, including CYP1A1 and CYP1A2, exhibit a high preference for estradiol 2-hydroxylation, but human CYP1B1 greatly favors 4-hydroxylation. Here we show that heterologous expression of the human, monkey, dog, rat, and mouse CYP1B1 enzymes yields active proteins that differ in their estradiol hydroxylation specificity. The monkey and dog orthologs, like the human enzyme, preferentially catalyze 4-hydroxylation, but the rat and mouse enzymes favor 2-hydroxylation. Analysis of the CYP1B1 sequences in light of these findings suggested that one residue, Val395 in human CYP1B1, could account for the differential hydroxylation specificities. In fact, mutation of this valine in human CYP1B1 to the leucine present in the rat enzyme produces a human enzyme that has the 2-hydroxylation specificity of the rat enzyme. The converse is true when the leucine in the rat enzyme is mutated to the human valine. The role of CYP1B1 in estradiol carcinogenicity thus depends on the identity of this single amino acid residue.


Neurological outcomes in patients transported to hospital without a prehospital return of spontaneous circulation after cardiac arrest.
As emergency medical services (EMS) personnel in Japan are not allowed to perform termination of resuscitation in the field, most patients experiencing an out-of-hospital cardiac arrest (OHCA) are transported to hospitals without a prehospital return of spontaneous circulation (ROSC). As the crucial prehospital factors for outcomes are not clear in patients who had an OHCA without a prehospital ROSC, we aimed to determine the prehospital factors associated with 1-month favorable neurological outcomes (Cerebral Performance Category scale 1 or 2 (CPC 1-2)). We analyzed the data of 398,121 adult OHCA patients without a prehospital ROSC from a prospectively recorded nationwide Utstein-style Japanese database from 2007 to 2010. The primary endpoint was 1-month CPC 1-2. The rate of 1-month CPC 1-2 was 0.49%. Multivariate logistic regression analysis indicated that the independent variables associated with CPC 1-2 were the following nine prehospital factors: (1) initial non-asystole rhythm (ventricular fibrillation (VF): adjusted odds ratio (aOR), 9.37; 95% confidence interval (CI), 7.71 to 11.4; pulseless ventricular tachycardia (VT): aOR, 8.50; 95% CI, 5.36 to 12.9; pulseless electrical activity (PEA): aOR, 2.75; 95% CI, 2.40 to 3.15), (2) age <65 years (aOR, 3.90; 95% CI, 3.28 to 4.67), (3) arrest witnessed by EMS personnel (aOR, 2.82; 95% CI, 2.48 to 3.19), (4) call-to-hospital arrival time <24 minutes (aOR, 2.58; 95% CI, 2.22 to 3.01), (5) arrest witnessed by any layperson, (6) physician-staffed ambulance, (7) call-to-response time <5 minutes, (8) prehospital shock delivery, and (9) presumed cardiac cause. When four crucial key factors (with an aOR >2.0 in the regression model: initial non-asystole rhythm, age <65 years, EMS-witnessed arrest, and call-to-hospital arrival time <24 minutes) were present, the rates of 1-month CPC 1-2 and 1-month survival were 16.1% and 23.2% in initial VF, 8.3% and 16.7% in pulseless VT, and 3.8% and 9.4% in PEA, respectively. In OHCA patients transported to hospitals without a prehospital ROSC, nine prehospital factors were significantly associated with 1-month CPC 1-2. Of those, four are crucial key factors: initial non-asystole rhythm, age <65 years, EMS-witnessed arrest, and call-to-hospital arrival time <24 minutes.


p16 Stimulates CDC42-dependent migration of hepatocellular carcinoma cells.
Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths worldwide. Tumor dissemination to the extra-hepatic region of the portal vein, lymph nodes, lungs or bones contributes to the high mortality seen in HCC; yet, the molecular mechanisms responsible for HCC metastasis remain unclear. Prior studies have suggested a potential link between accumulated cytoplasm-localized p16 and tumor progression. Here we report that p16 enhances metastasis-associated phenotypes in HCC cells - ectopic p16 expression increased cell migration in vitro, and lung colonization after intravenous injection, whereas knockdown of endogenous p16 reduced cell migration. Interestingly, analysis of p16 mutants indicated that the Cdk4 interaction domain is required for stimulation of HCC cell migration; however, knockdown of Cdk4 and Cdk6 showed that these proteins are dispensable for this phenomenon. Intriguingly, we found that in p16-positive HCC samples, p16 protein is predominantly localized in the cytoplasm. In addition, we identified a potential role for nuclear-cytoplasmic shuttling in p16-stimulated migration, consistent with the predominantly cytoplasmic localization of p16 in IHC-positive HCC samples. Finally, we determined that p16-stimulated cell migration requires the Cdc42 GTPase. Our results demonstrate for the first time a pro-migratory role for p16, and suggest a potential mechanism for the observed association between cytoplasmic p16 and tumor progression in diverse tumor types.


The primary structure of aspartate aminotransferase from pig heart muscle. Partial sequences determined by digestion with thermolysin and elastase.
Peptides produced by thermolytic digestion of aminoethylated aspartate aminotransferase and of the oxidized enzyme were isolated and their amino acid sequences determined. Digestion by elastase of the carboxymethylated enzyme gave peptides representing approximately 40% of the primary structure. Fragments from these digests overlapped with previously reported sequences of peptides obtained by peptic and tryptic digestion (Doonan et al., 1972), giving ten composite peptides containing 395 amino acid residues. The amino acid composition of these composite peptides agrees well with that of the intact enzyme. Confirmatory results for some of the present data have been deposited as Supplementary Publication 50018 at the National Lending Library for Science and Technology, Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1973) 131, 5.


A Difluoroboron β-Diketonate Probe Shows "Turn-on" Near-Infrared Fluorescence Specific for Tau Fibrils.
Tau aggregation in neuronal cells has recently received significant attention as a robust predictor of the progression of Alzheimer's disease (AD) because of its proven correlation with the degree of cognitive impairment in AD patients. Accordingly, noninvasive imaging of tau aggregates has been highlighted as a promising diagnostic tool for AD. We have previously identified a tau-specific "turn-on" near-infrared fluorescent (NIRF) probe (1), and, in this study, structural modification was performed to optimize its physicochemical as well as fluorescence properties. Thus, a series of fluorescent dyes (2a-2j) composed of a variously substituted difluoroboron β-diketonate and an N,N-dimethylaniline moiety linked by a length-extendable π-bridge were prepared. Among those, isobutyl-substituted difluoroboron β-ketonate with a π-conjugated 1,4-butadienyl linker (2e) showed the most promising properties as a tau-specific NIRF probe. Compared with 1, the "turn-on" fluorescence of 2e was more specific to tau fibrils, and it showed 8.8- and 6.2-times higher tau-over-Aβ and tau-over-BSA specificity, respectively. Also, the fluorescence intensity of 2e upon binding to tau fibrils was substantially higher (∼2.9 times) than that observed from 1. The mechanism for tau-specificity of 2e was investigated, which suggested that the molecular rotor-like property of 2e enables specific recognition of the microenvironment of tau aggregates to emit strong fluorescence. In transgenic cell lines stably expressing GFP-tagged tau proteins, 2e showed good colocalization with tau-GFP. Moreover, the fluorescence from 2e exhibited almost complete overlap with p-Tau antibody staining in the human AD brain tissue section. Collectively, these observations demonstrate the potential of 2e as a tau-specific fluorescent dye in both in vitro and ex vivo settings.


Host preference and zoonotic potential of Chlamydia psittaci and C. gallinacea in poultry.
Chlamydia psittaci and C. gallinacea are obligate intracellular bacteria infecting poultry. We conducted a survey in two poultry slaughterhouses that were processing either exclusively ducks (A) or various poultry species except ducks (B). Cloacal swabs were collected from all incoming poultry flocks in the course of a week, and blood samples and pharyngeal swabs were taken from workers. Swabs were examined using PCR and sera were analyzed with two immunoassays. PCR testing revealed the presence of C. psittaci in 9/38 duck flocks and the complete absence of C. gallinacea in these flocks (slaughterhouse A), whereas 16/33 Chlamydiaceae-positive poultry flocks handled in slaughterhouse B harbored C. gallinacea only. In an episode of psittacosis in slaughterhouse A, where one PCR-positive worker presented clinical signs, seroconversions were detected in 10 workers. In contrast, serological responses of slaughterhouse B workers to C. psittaci were generally low. This is in line with the almost complete absence of C. psittaci in handled flocks, where in additional sampling campaigns the agent was detected only once in the course of a year. Our study indicates that C. psittaci has a certain preference for ducks, whereas C. gallinacea was the predominant chlamydial agent in chickens and guinea fowl flocks.


Purinergic P2X7 receptor as a potential therapeutic target in depression.
The elaborate mechanisms of depression have always been a research hotspot in recent years, and the pace of research has never ceased. The P2X7 receptor (P2X7R) belongs to one of the adenosine triphosphates (ATP)-gated cation channels that exist widely in brain tissues and play a prominent role in the regulation of depression-related pathology. To date, the role of purinergic P2X7R in the mechanisms underlying depression is not fully understood. In this review, we conclude that the purinergic receptor P2X7 is a potential therapeutic target for depression based on research results published over the past 5 years in Google Scholar and the National Library of Medicine (PubMed). Additionally, we introduced the functional characteristics of P2X7R and confirmed that excessive activation of P2X7R led to increased release of inflammatory cytokines, which eventually contributed to depression. Furthermore, the inhibition of P2X7R produced antidepressant-like effects in animal models of depression, further proving that P2X7R signalling mediates depression-like behaviours. Finally, we summarised related studies on drugs that exert antidepressant effects by regulating the expression of P2X7R. We hope that the conclusions of this review will provide information on the role of P2X7R in the neuropathophysiology of depression and novel therapeutic targets for the treatment of depression.


Integrated Analysis of Ovarian Juvenile Granulosa Cell Tumors Reveals Distinct Epigenetic Signatures and Recurrent TERT Rearrangements.
Adult granulosa cell tumor (AGCT) is characterized by the somatic FOXL2 p.C134W mutation, and recurrences have been associated with TERT promoter and KMT2D-truncating mutations. Conversely, the molecular underpinnings of the rare juvenile granulosa cell tumor (JGCT) have not been well elucidated. To this end, we applied a tumor-only integrated approach to investigate the genomic, transcriptomic, and epigenomic landscape of 31 JGCTs to identify putative oncogenic drivers. Multipronged analyses of 31 JGCTs were performed utilizing a clinically validated next-generation sequencing (NGS) panel targeting 580 cancer-related genes for genomic interrogation, in addition to targeted RNA NGS for transcriptomic exploration. Genome-wide DNA methylation profiling was conducted using an Infinium Methylation EPIC array targeting 866,562 CpG methylation sites. We identified frequent KMT2C-truncating mutations along with other mutated genes implicated in the switch/sucrose nonfermentable (SWI/SNF) chromatin remodeling complex, in addition to previously reported hotspot AKT1 and DICER1 mutations. Targeted transcriptome sequencing revealed recurrent TERT rearrangements (13%) involving partners CLPTM1L or DROSHA, and differential gene expression analysis showed FGFR1 upregulation in the TERT non-rearranged JGCTs under direct promoter control. Genome-wide DNA methylation rendered a clear delineation between AGCTs and JGCTs at the epigenomic level, further supporting its diagnostic utility in distinguishing among these tumors. This is the largest comprehensive molecular study of JGCTs, where we further expand our current understanding of JGCT pathogenesis and demonstrate putative oncogenic drivers and TERT rearrangements in a subset of tumors. Our findings further offer insights into possible targeted therapies in a rare entity.


[Certain humoral factors in acute myocardial infarct complicated by cardiogenic shock].
The activity of the sympathoadrenal and renin-angiotensin-aldosterone systems was studied in patients with myocardial infarction complicated by cardiogenic shock. During the initial 12 hours after the onset of myocardial infarction an increased plasma level of adrenalin and noradrenal is observed, which then returns to the normal level on the 3rd day. In cases of cardiogenic shock a lower content of catecholamines in blood is noted. The determinations of renin activity in the plasma of peripheral venous blood demonstrated that it was increased since the 1st day of the disease in the myocardial infarction patients, but that its maximum elevation fell within the 7th--10th days. In patients with cardiogenic shock the plasma renin activity was much higher than in those with uncomplicated myocardial infarction. Urine aldosterone excretion was higher in patients with cardiogenick shock since the 1st day of the disease than in those with an uncomplicated course of myocardial infarction. The study of the electrolyte balance in patients with myocardial infarction complicated by cardiogenic shock demonstrated that on the 1st day of the disease the potassium level in plasma and erythrocytes was decreased, while the sodium concentration was somewhat increased.


TELOMERASE ACTIVATOR1 induces telomerase activity and potentiates responses to auxin in Arabidopsis.
Telomerase activity is highly regulated, abundant in rapidly dividing cells and reproductive organs, but undetectable in most other differentiated tissues. Little is known about mechanisms that regulate telomerase. Here, we used a biochemical assay to screen activation-tagged lines of Arabidopsis thaliana for mutants that ectopically express this enzyme in their leaves. In one such mutant, a previously uncharacterized zinc-finger protein we designate TELOMERASE ACTIVATOR1 (TAC1) is overexpressed and induces telomerase in fully differentiated leaves without stimulating progression through the cell cycle. Reducing endogenous concentrations of auxin in the mutant blocks the ability of TAC1 to induce telomerase. This result, along with other phenotypes of the mutant, such as the ability of cells to grow in culture without exogenous auxin and increased sensitivity of primary root growth to exogenous auxin, indicates that TAC1 not only is part of the previously reported link between auxin and telomerase expression but also potentiates other classic responses to this phytohormone.


[Wide-angle x-ray scattering comparison of the structure of crystalline cytochrome c and cytochrome c in solution].
Large-angle X-ray diffuse scattering has been used for studying the conformational changes in cytochrome c during its transition from crystal into solution and during a change of the electron state of the heme. It has been found that the structure of cytochrome c in solution differs from its structure in crystal by a shift of the chain fragment in the region of 60-77 amino acid residues. The studies of the oxidized, reduced and cyanoforms of protein in solution have not revealed noticeable changes in the protein structure.


Implementation and Operational Research: Expedited Results Delivery Systems Using GPRS Technology Significantly Reduce Early Infant Diagnosis Test Turnaround Times.
The objective of this study was to quantify the impact of a new technology to communicate the results of an infant HIV diagnostic test on test turnaround time and to quantify the association between late delivery of test results and patient loss to follow-up. We used data collected during a pilot implementation of Global Package Radio Service (GPRS) printers for communicating results in the early infant diagnosis program in Mozambique from 2008 through 2010. Our dataset comprised 1757 patient records, of which 767 were from before implementation and 990 from after implementation of expedited results delivery system. We used multivariate logistic regression model to determine the association between late result delivery (more than 30 days between sample collection and result delivery to the health facility) and the probability of result collection by the infant's caregiver. We used a sample selection model to determine the association between late result delivery to the facility and further delay in collection of results by the caregiver. The mean test turnaround time reduced from 68.13 to 41.05 days post-expedited results delivery system. Caregivers collected only 665 (37.8%) of the 1757 results. After controlling for confounders, the late delivery of results was associated with a reduction of approximately 18% (0.44 vs. 0.36; P < 0.01) in the probability of results collected by the caregivers (odds ratio = 0.67, P < 0.05). Late delivery of results was also associated with a further average increase in 20.91 days of delay in collection of results (P < 0.01). Early infant diagnosis program managers should further evaluate the cost-effectiveness of operational interventions (eg, GPRS printers) that reduce delays.


Controlling the False Discovery Rate for Feature Selection in High-resolution NMR Spectra.
Successful implementation of feature selection in nuclear magnetic resonance (NMR) spectra not only improves classification ability, but also simplifies the entire modeling process and, thus, reduces computational and analytical efforts. Principal component analysis (PCA) and partial least squares (PLS) have been widely used for feature selection in NMR spectra. However, extracting meaningful metabolite features from the reduced dimensions obtained through PCA or PLS is complicated because these reduced dimensions are linear combinations of a large number of the original features. In this paper, we propose a multiple testing procedure controlling false discovery rate (FDR) as an efficient method for feature selection in NMR spectra. The procedure clearly compensates for the limitation of PCA and PLS and identifies individual metabolite features necessary for classification. In addition, we present orthogonal signal correction to improve classification and visualization by removing unnecessary variations in NMR spectra. Our experimental results with real NMR spectra showed that classification models constructed with the features selected by our proposed procedure yielded smaller misclassification rates than those with all features.


[A model of regulation of crossing over].
Very high variability is characteristic of crossing over. A mechanism of crossing-over variation is proposed on the basis of new data obtained by Chadov et al. Variations are believed to be determined by the character of pairing preceding crossing over and the mechanism of transforming contacts into exchanges. Pairing of homologues is assumed to involve consecutive formation of local contacts between them. The onset of this process depends of the chromosome structure. Exchanges appear in contact sites. Determination of crossing-over sites occurs when the contact number in a meiocyte reaches a certain threshold. The number of contacts formed in each chromosome pair by this time can vary. This results in variations of all parameters of crossing over: recombinational length (RL), exchange number per chromosome arm, and interference. The effect of a chromosome aberration on crossing over in aberrant and heterologous chromosome pairs, i.e., the interchromosomal effect on crossing over (IEC) and the mechanisms determining exchange number and interference, are explained on the basis of the proposed model.


In vivo studies on lysosubtilin. I. Efficacy for prophylaxis and treatment of gastrointestinal disorders in newborn calves.
This is the first part of a research report on the in vivo properties of lysosubtilin, a broad-spectrum preparation of lytic enzymes from Bacillus subtilis designed for veterinary medicine. This preparation has both antimicrobial properties and the capacity to compensate for digestive enzyme deficiencies in newborn calves. This study demonstrates its efficacy for both the prophylaxis and treatment of calf gastrointestinal disorders. Dose determination studies involved calves fed with colostrum or later milk, supplemented with 10 g/L of sodium bicarbonate, which received either a prophylactic dose of lysosubtilin of 2 x 10(4) U/kg of weight given twice daily for 10 days beginning with the first feeding (30-45 min after birth) or a therapeutic dose of 2.5 x 10(4) U/kg of weight given twice daily until recovery. These could be used for the prophylaxis and treatment, respectively, of the disorders mentioned above with an efficacy up to 100%. Trials in veterinary medicine were carried out in four former Soviet Union republics. More than 16,000 newborn calves were involved. The efficacy of prophylaxis and treatment was found to be 94.4-100% and 95.7-100% respectively, while the efficacy of regular methods was no higher than 82.0 and 91.8%, respectively. A positive impact of lysosubtilin on the improvement of digestive tract functioning, development of an animal and its protective functions against disease was also demonstrated. The consequent increases in calves' daily weight gain, abomasal proteolytic capacity as well as in amount of gamma-globulin, lysozyme and bactericidal activities in their blood sera were significant.


Increased cortical recruitment in Huntington's disease using a Simon task.
Cognitive deficits in Huntington's disease (HD) have been attributed to neuronal degeneration within the striatum; however, postmortem and structural imaging studies have revealed more widespread morphological changes. To examine the impact of HD-related changes in regions outside the striatum, we used functional magnetic resonance imaging (fMRI) in HD to examine brain activation patterns using a Simon task that required a button press response to either congruent or incongruent arrow stimuli. Twenty mild to moderate stage HD patients and 17 healthy controls were scanned using a 3T GE scanner. Data analysis involved the use of statistical parametric mapping software with a random effects analysis model to investigate group differences brain activation patterns compared to baseline. HD patients recruited frontal and parietal cortical regions to perform the task, and also showed significantly greater activation, compared to controls, in the caudal anterior cingulate, insula, inferior parietal lobules, superior temporal gyrus bilaterally, right inferior frontal gyrus, right precuneus/superior parietal lobule, left precentral gyrus, and left dorsal premotor cortex. The significantly increased activation in anterior cingulate-frontal-motor-parietal cortex in HD may represent a primary dysfunction due to direct cell loss or damage in cortical regions, and/or a secondary compensatory mechanism of increased cortical recruitment due to primary striatal deficits.


[The evaluation of complications following augmentation rhinoplasty with silicon gel].
Since 1985, 15 patients who suffered from complications after augmentation rhinoplasty by injection of silicone gel have been reoperated on in our department. These complications included: (1) unsatisfactory aesthetic result in nine cases; (2) ulceration in three cases; (3) ophthalmic complications, including blindness of one-eye (2 cases) and two-eyes with severe vision disorders (1 case). The reasons for unsatisfactory aesthetic result and other complications were analysed and evaluated.


Childhood asthma is a risk factor for the development of chronic obstructive pulmonary disease.
This review will catalog the many recent longitudinal studies that have investigated the relationship between asthma and lung function, or the persistence and trajectories of lung function deficits. Recent work has reported on 50-year follow-ups of some prominent population cohorts. A history of asthma confers a 10-30-fold risk of chronic obstructive pulmonary disease. Individuals reaching a reduced maximum growth of forced expiratory volume in 1 s in early adulthood are at risk for early or more severe chronic obstructive pulmonary disease (COPD). Taken together, there is a wealth of overlapping cohort studies of lung function, asthma and COPD. These show that asthma is associated with reduced lung function, which may start in infancy or prenatally, persists through childhood and adulthood and predisposes for early or more severe COPD.


Parallel Independent Losses of G-Type Lysozyme Genes in Hairless Aquatic Mammals.
Lysozyme enzymes provide classic examples of molecular adaptation and parallel evolution, however, nearly all insights to date come from chicken-type (c-type) lysozymes. Goose-type (g-type) lysozymes occur in diverse vertebrates, with multiple independent duplications reported. Most mammals possess two g-type lysozyme genes (Lyg1 and Lyg2), the result of an early duplication, although some lineages are known to have subsequently lost one copy. Here we examine g-type lysozyme evolution across >250 mammals and reveal widespread losses of either Lyg1 or Lyg2 in several divergent taxa across the mammal tree of life. At the same time, we report strong evidence of extensive losses of both gene copies in cetaceans and sirenians, with an additional putative case of parallel loss in the tarsier. To validate these findings, we inspected published short-read data and confirmed the presence of loss of function mutations. Despite these losses, comparisons of selection pressures between intact g- and c-type lysozyme genes showed stronger purifying selection in the former, indicative of conserved function. Although the reasons for the evolutionary loss of g-type lysozymes in fully aquatic mammals are not known, we suggest that this is likely to at least partially relate to their hairlessness. Indeed, although Lyg1 does not show tissue-specific expression, recent studies have linked Lyg2 expression to anagen hair follicle development and hair loss. Such a role for g-type lysozyme would explain why the Lyg2 gene became obsolete when these taxa lost their body hair.


SIRT2 overexpression in hepatocellular carcinoma mediates epithelial to mesenchymal transition by protein kinase B/glycogen synthase kinase-3β/β-catenin signaling.
Sirtuin 1 (SIRT1) has been implicated in telomere maintenance and the growth of hepatocellular carcinoma (HCC). Nevertheless, the role of other sirtuins in the pathogenesis of HCC remains elusive. We found that sirtuin 2 (SIRT2), another member of the sirtuin family, also contributes to cell motility and invasiveness of HCC. SIRT2 is up-regulated in HCC cell lines and in a subset of human HCC tissues (23/45). Up-regulations of SIRT2 in primary HCC tumors were significantly correlated with the presence of microscopic vascular invasion (P = 0.001), a more advanced tumor stage (P = 0.004), and shorter overall survival (P = 0.0499). Functional studies by short hairpin RNA-mediated suppression of SIRT2 expression in HCC cell lines revealed significant inhibition of motility and invasiveness. Depletion of SIRT2 also led to the regression of epithelial-mesenchymal transition (EMT) phenotypes, whereas the ectopic expression of SIRT2 in the immortalized hepatocyte cell line L02 promoted cell motility and invasiveness. Mechanistic studies revealed that SIRT2 regulates the deacetylation and activation of protein kinase B, which subsequently impinges on the glycogen synthase kinase-3β/β-catenin signaling pathway to regulate EMT. Our findings have uncovered a novel role for SIRT2 in HCC metastasis, and provide a rationale to explore the use of sirtuin inhibitors in HCC therapy. (HEPATOLOGY 2013;).


Ambulatory Goeckerman treatment of psoriasis: experience with 200 patients.
At a psoriasis day care centre 200 patients were treated with an ambulatory Goeckerman regimen, 25 of them twice, because of recurrence. This treatment consists of the application of a coal tar preparation at home at bedtime, followed the next day by exposure to high-intensity short wavelength ultraviolet radiation (UVB) at the centre. Treatment was given 5 days a week for 1 month. The psoriasis cleared in 86% of the patients after a mean of 21.6 UVB treatment sessions. The mean length of remission was 5.1 months, but at the time of follow-up 15 patients were still free of psoriasis. Compliance with the regimen was good to excellent in 94% of the patients. In our hands ambulatory treatment of psoriasis is much less expensive than hospital treatment and gives better results than photochemotherapy.


Anatomical remodelling of the supraoptic nucleus: changes in synaptic and extrasynaptic transmission.
The adult hypothalamic-neurohypophysial system undergoes activity-dependent morphological plasticity that modifies the astrocytic enwrapping of its magnocellular neurones. For a long time, the functional consequences of such changes have remained hypothetical. Modifications in the glial environment of neurones are expected to have important physiological repercussions in view of the various functions played by astrocytes in the central nervous system. In particular, glial cells are essential for uptake of neurotransmitters, including glutamate, and for physically and functionally restricting diffusion of neuroactive substances within the extracellular space. Recent studies performed in the supraoptic nucleus of lactating and chronically dehydrated animals, in conditions where astrocytic coverage of neurones is reduced, have revealed a significant impairment of glutamate clearance. The resulting accumulation of the excitatory amino acid in the extracellular space around glutamatergic inputs causes an enhanced activation of presynaptic metabotropic glutamate receptors that inhibit transmitter release. In the supraoptic nucleus of lactating rats, neuroglial remodelling is accompanied by modification of the geometry, size and diffusion properties of the extracellular space. The latter observations suggest that, in the activated supraoptic nucleus, the range of action and the concentration of released neuroactive substances may be significantly enhanced. Overall, our observations indicate that the glial environment of supraoptic neurones influences synaptic glutamatergic transmission, as well as extrasynaptic forms of communication.


Clinical course and prognosis of musculoskeletal pain in patients referred for physiotherapy: does pain site matter?
Danish patients with musculoskeletal disorders are commonly referred for primary care physiotherapy treatment but little is known about their general health status, pain diagnoses, clinical course and prognosis. The objectives of this study were to 1) describe the clinical course of patients with musculoskeletal disorders referred to physiotherapy, 2) identify predictors associated with a satisfactory outcome, and 3) determine the influence of the primary pain site diagnosis relative to those predictors. This was a prospective cohort study of patients (n = 2,706) newly referred because of musculoskeletal pain to 30 physiotherapy practices from January 2012 to May 2012. Data were collected via a web-based questionnaire 1-2 days prior to the first physiotherapy consultation and at 6 weeks, 3 and 6 months, from clinical records (including primary musculoskeletal symptom diagnosis based on the ICPC-2 classification system), and from national registry data. The main outcome was the Patient Acceptable Symptom State. Potential predictors were analysed using backwards step-wise selection during longitudinal Generalised Estimating Equation regression modelling. To assess the influence of pain site on these associations, primary pain site diagnosis was added to the model. Of the patients included, 66% were female and the mean age was 48 (SD 15). The percentage of patients reporting their symptoms as acceptable was 32% at 6 weeks, 43% at 3 months and 52% at 6 months. A higher probability of satisfactory outcome was associated with place of residence, being retired, no compensation claim, less frequent pain, shorter duration of pain, lower levels of disability and fear avoidance, better mental health and being a non-smoker. Primary pain site diagnosis had little influence on these associations, and was not predictive of a satisfactory outcome. Only half of the patients rated their symptoms as acceptable at 6 months. Although satisfactory outcome was difficult to predict at an individual patient level, there were a number of prognostic factors that were associated with this outcome. These factors should be considered when developing generic prediction tools to assess the probability of satisfactory outcome in musculoskeletal physiotherapy patients, because the site of pain did not affect that prognostic association.