Defining a new testosterone threshold for medical castration: Results from a prospective cohort series.
We seek to determine if testosterone levels below the accepted castration threshold (50 ng/dL) have an impact on time to progression to castrate-resistant prostate cancer (CRPC). This is a prospective cohort series of patients undergoing androgen deprivation therapy (ADT) with luteinizing hormone-releasing hormone agonist or antagonist at a tertiary centre from 2006 to 2011. Serum testosterone level was assessed every 3 months. Patients with any testosterone >50 ng/dL were excluded. Patients were stratified into groups based on those achieving mean testosterone levels <20 ng/dL and <32 ng/dL. Progression to CRPC was assessed with the Kaplan-Meier method and compared with the log-rank test. A total of 32 patients were included in this study. Mean patient follow-up was 25.7 months. Patients with a 9-month serum testosterone <32 ng/dL had a significantly increased time to CRPC compared to patients with testosterone 32 to 50 ng/dL (p = 0.001, median progression-free survival (PFS) 33.1 months [<32 ng/dL] vs. 12.5 months [>32 ng/dL]). Patients with first year mean testosterone <32 ng/dL also had a significantly increased time to CRPC compared to 32 to 50 ng/dL (p = 0.05, median PFS 33.1 months [<32 ng/dL] vs. 12.5 months [32-50 ng/dL]). A testosterone <20 ng/dL compared to 20 to 50 ng/dL did not significantly predict with time to CRPC. This study supports a lower testosterone threshold to define optimal medical castration (T <32 ng/dL) than the previously accepted standard of 50 ng/dL. Testosterone levels during ADT serve as an early predictor of disease progression and thus should be measured in conjunction with prostate-specific antigen.


Monitoring of population shifts in an enriched nitrifying system under gradually increased cadmium loading.
The changes in nitrifying bacterial population under cadmium loading were monitored and evaluated in a laboratory scale continuous-flow enriched nitrification system. For this purpose, the following molecular microbiological methods were used: slot-blot hybridization, denaturing gradient gel electrophoresis (DGGE), real-time PCR followed by melting curve analysis, cloning and sequence analysis. The initial cadmium concentration was incrementally increased from 1 to 10mg/l which led to a drop in ammonia removal efficiency from 99 to 10%. Inhibition was recovered when cadmium loading was stopped. During the second application of cadmium, nitrifying population became more tolerant. Even at 15mg/l Cd, only a minor inhibition was observed. To investigate the variations in ammonia and nitrite oxidizing bacteria populations in a period of 483 days, ammonia monooxygenase (amoA) and 16S rRNA genes-based molecular techniques were used. An obvious shift was experienced in the diversity of ammonia oxidizers after the first application of 10mg/l Cd. Metal-tolerant ammonia oxidizing species became dominant and the microbial diversity sharply shifted from Nitrosomonas and Nitrosococcus sp. to Nitrosospira sp. which were observed to tolerate higher cadmium loadings. This result indicated that the extent of nitrification inhibition was not only related to the metal concentration and quantity of microorganisms but also depended on the type of species.


[Sandflies (Diptera: Psychodidae) from a cerrado area of Corumbataí Municipality, Central East Region of São Paulo State, Brazil].
Distribution and ecology knowledge of sandfly species is essential for epidemiology vigilance and risk determination for transmission of leishmaniasis. Sandfly trapping was carried out in a cerrado strictu sensu vegetation fragment in a rural area of Corumbataí Municipality, São Paulo State, Brazil, during July to November 2004. Two CDC light traps were used from 18h to 8h, once a month, resulting in 112 total hours of capture. During this period, 60 sandfly specimens of ten different species were sampled. The most abundant species and the one found in all captures was Pintomyia monticola totalizing 15 (25.0%) specimens, the second more abundant one was P. pessoai with 14 (23.3%) and the third one was Brumptomyia guimaraesi with 12 (20.0%) individuals collected. Other captured species were Psathyromyia aragaoi, B. avellari, B. brumpti, B. cunhai, P. bianchigalatiae, Evandromyia termitophila and Nyssomyia whitmani which corresponded to 19 (31.2%) specimens captured. Pintomyia pessoai and N. whitmani presence indicates transmission risk of American Cutaneous Leishmaniasis in the area.


Condom use and HIV-related behaviors in urban Tanzanian men who have sex with men: a study of beliefs, HIV knowledge sources, partner interactions and risk behaviors.
Although studies have been conducted in neighboring countries, there are no published data on men who have sex with men (MSM) in mainland Tanzania. We report on a respondent-driven sampling study of 271 MSM in Dar es Salaam, Tanzania. The sample covered a wide range of educational attainment and employment, median age was 24, and all respondents had heard of HIV/AIDS, mostly through public media. Those satisfied with media information on HIV were younger, had lower education, and had obtained their information from health facilities. Over two-thirds believed that having one faithful partner and using condoms would protect against HIV: nevertheless, more than two-thirds were worried about HIV infection. Two-thirds had had a relationship with a woman, one-third in the past year. Predictors of non-use of condoms for anal sex with last casual partner were younger age, not being worried about HIV infection, and agreeing to have sex even if a condom was refused. There was no significance in proportion using a condom with last casual (43%) and last regular (49%) partner. Most partners (MSM knew a median of 10 other MSM) were met in bars, music halls, and in the home/local environment, and 70% of MSM described their sexual position as "bottom." Sixty percent reported having an HIV test and the great majority was comfortable discussing condoms with partners and friends: half would refuse to have sex if condoms were not agreed to. These data suggest a significant "gay" community in Dar es Salaam with relatively accurate HIV information but moderate condom use, HIV testing and ability to refuse unsafe sex. There is clearly scope for targeted HIV prevention programs in the MSM community in Tanzania.


[Activities at an Infectious Disease Intensive Care Unit (1988-1997)].
The practice of an Infectious Diseases Intensive Care Unit for a period of ten years (1988-1997) is reviewed. In this time 1,191 patients were treated--822 had more than 15 years of age and 369 were younger. The occupation rate ranged between 74.4% and 90.8%, and the mean patient stay in the UCI ranged between 6.7 and 12.5 days. Nine hundred and forty-four patients were discharged cured or improved during the stay and 247 (20.7%) died. The most frequent pathologies were: meningitis--371 patients (33.6%); encephalitis--103 (8.6%); and tetanus 143 (12.0%) patients. The morbidity and mortality of different pathologies admitted to the Intensive Care Unit are presented. The mortality rate of the two most common diseases (meningitis and tetanus) is discussed. The need for Intensive Care in the field of Infectious Diseases is stressed.


Sensorimotor synchronisation with higher metrical levels in music shortens perceived time.
The aim of the present study was to investigate if the perception of time is affected by actively attending to different metrical levels in musical rhythmic patterns. In an experiment with a repeated-measures design, musicians and non-musicians were presented with musical rhythmic patterns played at three different tempi. They synchronised with multiple metrical levels (half notes, quarter notes, eighth notes) of these patterns using a finger-tapping paradigm and listened without tapping. After each trial, stimulus duration was judged using a verbal estimation paradigm. Results show that the metrical level participants synchronised with influenced perceived time: actively attending to a higher metrical level (half notes, longer inter-tap intervals) led to the shortest time estimations, hence time was experienced as passing more quickly. Listening without tapping led to the longest time estimations. The faster the tempo of the patterns, the longer the time estimation. While there were no differences between musicians and non-musicians, those participants who tapped more consistently and accurately (as analysed by circular statistics) estimated durations to be shorter. Thus, attending to different metrical levels in music, by deliberately directing attention and motor activity, affects time perception.


A novel capsid-XL32-derived adeno-associated virus serotype prompts retinal tropism and ameliorates choroidal neovascularization.
Gene therapy has been adapted, from the laboratory to the clinic, to treat retinopathies. In contrast to subretinal route, intravitreal delivery of AAV vectors displays the advantage of bypassing surgical injuries, but the viral particles are more prone to be nullified by the host neutralizing factors. To minimize such suppression of therapeutic effect, especially in terms of AAV2 and its derivatives, we introduced three serine-to-glycine mutations, based on the phosphorylation sites identified by mass spectrum analysis, to the XL32 capsid to generate a novel serotype named AAVYC5. Via intravitreal administration, AAVYC5 was transduced more effectively into multiple retinal layers compared with AAV2 and XL32. AAVYC5 also enabled successful delivery of anti-angiogenic molecules to rescue laser-induced choroidal neovascularization and astrogliosis in mice and non-human primates. Furthermore, we detected fewer neutralizing antibodies and binding IgG in human sera against AAVYC5 than those specific for AAV2 and XL32. Our results thus implicate this capsid-optimized AAVYC5 as a promising vector suitable for a wide population, particularly those with undesirable AAV2 seroreactivity.


Bilateral symmetry and coherence of subthalamic nuclei beta band activity in Parkinson's disease.
Abnormal synchronization of neuronal activity in the basal ganglia has been associated with the dysfunction of sensorimotor circuits in Parkinson's disease (PD). In particular, oscillations at frequencies within the beta range (13-35 Hz) are specifically modulated by dopaminergic medication and are correlated with the clinical state of the subjects. While these oscillations have been shown to be coherent ipsilaterally within the basal ganglia and between the basal ganglia nuclei and the ipsilateral motor cortex in PD, the bilateral extent of their coherence has never been characterized. Here we demonstrate for the first time that the beta band oscillations recorded in the local field potential of the subthalamic nuclei (STN), while appearing different across subjects, are occurring at the same frequencies bilaterally (p<0.001) and are coherent between the two STNs of individual PD subjects (11/12 cases, p<0.05). These findings suggest the existence of a bilateral network controlling the beta band activity in the basal ganglia in PD.


Strain variation in susceptibility to monoclonal antibody-induced transplantation tolerance.
We have reproducibly induced specific tolerance to multiple minor histocompatibility antigens with nondepleting anti-CD4 and -CD8 monoclonal antibodies. The tolerance induced is effective for the lifetime of the host. We have tested this therapy in a number of mouse strain combinations to further understand the mechanisms. Various mouse strains were grafted with allogeneic tail skin with and without nondepleting CD4- and CD8-specific monoclonal antibody therapy. The grafts were monitored daily for signs of rejection. Whereas the CBA/Ca (H2k) strain can be made tolerant to skin grafts that are mismatched at multiple minor histocompatibility antigens indefinitely, using the same protocol, long-term survival of similarly mismatched grafts on the HW80 (B6 congenic for BALB H1) mouse strain is limited to around 8 weeks. Interestingly, the B10.BR strain, which is also of the H2k haplotype, is also not readily tolerized. In addition, an F1 between the CBA/Ca and the resistant B10.BR strains is B10.BR-like in its susceptibility to tolerance induction. Susceptibility to such antibody-dependent tolerance induction is not related to immunogenicity because grafts mismatched at only a single minor antigen also do not reproducibly survive beyond 8 weeks when grafted onto HW80 mice in the presence of the antibody therapy. The data strongly suggest that the B6/B10 genetic background confers a level of resistance to CD4- and CD8-specific monoclonal antibody-dependent tolerance induction.


Extending conceptual frameworks: life course epidemiology for the study of back pain.
Epidemiological studies have identified important causal and prognostic factors for back pain, but these frequently only identify a proportion of the variance, and new factors add little to these models. Recently, interest has increased in studying diseases over the life course, stimulated by the 1997 book by Kuh and Ben-Shlomo, a move accompanied by important conceptual and methodological developments. This has resulted in improvements in the understanding of other conditions like cardiovascular and respiratory disease. This paper aims to examine how conceptual frameworks from life course epidemiology could enhance back pain research. Life course concepts can be divided into three categories. Concept 1: patterns over time, risk chains and accumulation. Simple 'chains of risk' have been studied - e.g. depression leading to back pain - but studies involving more risk factors in the chain are infrequent. Also, we have not examined how risk accumulation influences outcome, e.g. whether multiple episodes or duration of depression, throughout the life course, better predicts back pain. One-year back pain trajectories have been described, and show advantages for studying back pain, but there are few descriptions of longer-term patterns with associated transitions and turning points. Concept 2: influences and determinants of pathways. Analyses in back pain studies commonly adjust associations for potential confounders, but specific analysis of factors modifying risk, or related to the resilience or susceptibility to back pain, are rarely studied. Concept 3: timing of risk. Studies of critical or sensitive periods - crucial times of life which influence health later in life - are scarce in back pain research. Such analyses could help identify factors that influence the experience of pain throughout the life course. Back pain researchers could usefully develop hypotheses and models of how risks from different stages of life might interact and influence the onset, persistence and prognosis of back pain throughout the life course. Adoption of concepts and methods from life course epidemiology could facilitate this.


Hepatitis C virus infection in hemodialysis patients is not associated with insulin resistance, inflammation and atherosclerosis.
Hepatitis C virus (HCV) infection is associated with increased levels of inflammatory markers and is also a significant risk factor for the development of Type 2 diabetes mellitus (DM) in the general population. In attempt to address this issue in chronic hemodialysis (HD) patients, we evaluated the relation of HCV infection with inflammatory markers, endothelial dysfunction, insulin resistance and atherosclerosis. In a cohort of 72 HD patients (36 (50%) had a positive HCV EIA test and 36 (50%) had a negative anti-HCV test), we examined HCV antibody status, insulin resistance (HOMA-IR) and beta-cell function (HOMA-beta), serum inflammatory parameters (high sensitive C-reactive protein (CRP), fibrinogen and procalcitonin), and performed echocardiography, high-resolution brachial artery ultrasound and B-mode carotid Doppler examination to assess the vascular functions and atherosclerosis. There were no differences in age, gender, body mass index (BMI), primary disease, duration of dialysis, smoking status, laboratory parameters except glucose, total cholesterol and LDL cholesterol between anti-HCV-positive and anti-HCV-negative groups. The serum concentrations of glucose, total cholesterol and LDL cholesterol were significantly lower in the anti-HCV-positive patients than anti-HCV-negative patients (p = 0.04, p = 0.02, p = 0.01, respectively). There were no significant differences in inflammatory parameters, total insulin secretion (HOMA-beta, p = 0.76) and insulin resistance (HOMA-R, p = 0.91) between anti-HCV-positive and negative patients. The intima media thickness, carotid plaque score and brachial artery endothelium-dependent dilatation did not differ significantly between the two groups (p = 0.44, 0.45, and 0.17, respectively). HCV infection was not related to atherosclerosis and insulin resistance in hemodialysis patients. Since hemodialysis patients had a large number of uremia-related cardiovascular risk factors, the effect of HCV infection could disappear in this group of patients.


Endoglin (CD105) expression in angiogenesis of primary hepatocellular carcinomas: analysis using tissue microarrays and comparisons with CD34 and VEGF.
Few studies about angiogenesis in hepatocellular carcinoma (HCC) have been conducted and little is known about the significance of angiogenesis in HCC. In this study, the clinicopathological significance of tumor microvessel density (MVD) was assessed in 105 patients with HCC by immunohistochemical staining of CD105, CD34, and vascular endothelial growth factor (VEGF). Moreover, the use of the tissue microarray technique in evaluating angiogenesis of HCC was appraised. The MVD by CD105 immunostaining (MVD-CD105) was significantly lower in larger tumors (5 cm diameter as a cutoff point, p=0.001), more aggressive tumors, as indicated by venous infiltration (present vs absent, p=0.001), and tumors with advanced TNM stage (stage I & II vs stage III, p=0.011). A lower score of MVD by CD34 immunostaining (MVD-CD34) showed significant association only with venous invasion (p<0.001), whereas the MVD by CD105 immunostaining in tissue microarray (MVD-MA) was significantly lower only in larger sized tumors (p=0.043). Moreover, MVD-CD105 was positively associated with the expression intensity of VEGF (p=0.009), but not for MVD-CD34 (p=0.088). When median scores of MVD were used as cut-off points, the patients with higher score of MVD-CD105 had a significantly poorer prognosis in either disease-free or overall survival analysis (p=0.002 and p=0.009, respectively), whereas similar prognostic significance of MVD-CD34 was not observed in overall survival analysis (p=0.052) but was observed in disease-free survival analysis (p=0.022). No prognostic significance of MVD-MA was found in either disease-free or overall survival analysis (p=0.277 and p=0.712, respectively). These data demonstrate the superiority of CD105 over CD34 as a marker of angiogenesis in HCC and indicate that the tissue microarray technique is unsuitable for evaluating angiogenesis in HCC.


Effect of cDMEM media containing Ectoine on human periodontal ligament mesenchymal stem cell survival and differentiation.
Ectoine is an amino acid that can preserve osmotic balance and has more preservative activity than other osmoregulators. There are no published reports on the osmoregulators' effects on viability or differentiation of dental stem cells. The aim of this study was to investigate the effect of Ectoine as a storage media on the viability and differentiation potential of human periodontal ligament mesenchymal stem cells (hPDLMSCs). hPDLMSCs were obtained from impacted third molar teeth. The cells were isolated, submitted to trilineage differentiation, and characterized by flow cytometer (FC). hPDLMSCs were incubated with different media containing Ectoine, complete DMEM (cDMEM), Ectoine+cDMEM, milk, and tap water for 2, 6, 12, and 24 h. The cells were analyzed by FC for viability, early-apoptosis, late apoptosis, and necrosis rates. Osteogenic and fibroblastic differentiation in hPDLMSCs were investigated by Alizarin red stain and vimentin expression. All treated groups showed significant decreases in cell viability after 2 h. Significant increases were detected in the number of dead cells between 2 and 12 h in all groups except the Ectoine+cDMEM group. The deposition of mineral matrix nodules was significantly higher in cells cultured with Ectoine+cDMEM compared with the other media. Higher vimentin expressions were detected in cells cultured with cDMEM and Ectoine+cDMEM media, respectively. Ectoine added to cDMEM media, promoted cell survival plus osteogenic and fibroblastic differentiation of hPDLMSCs.


Characterization of cytochrome P450 expression in human oesophageal mucosa.
The expression of cytochrome (CYP) P450 enzymes in human oesophageal mucosa was investigated in a total of 25 histologically non-neoplastic surgical tissue specimens by using specific antibodies in immunoblots and by RT-PCR mRNA analysis. The presence of CYP1A, 2E1, 3A and 4A enzymes was demonstrated by both techniques; CYP2A reactive protein was also detected by immunoblot. The presence of CYP4B1 mRNA was established but no specific antibody was available for detection of the corresponding protein by immunoblot. CYP2B6/7 mRNA was not detected in any sample. The mRNA transcripts for CYP1A1, 2E1, 4A11 and 4B1 were consistently detected in the majority of samples (>84%), whereas CYP1A2 mRNA was only detected in 11 of 19 specimens examined. An RT-PCR method to differentiate CYP3A4 and 3A5 mRNA was developed. This demonstrated CYP3A5 mRNA expression in all samples tested, whereas CYP3A4 mRNA was not detectable, suggesting that CYP3A5 is the major CYP3A protein in human oesophagus. There were significant interindividual variations in the amount of proteins, ranging from 8-fold for CYP4A to 43-fold for CYP2E1. For each patient, data on exposure to risk factors for oesophageal cancer were available, including tobacco smoke, alcohol, gastro-oesophageal reflux and hot beverage consumption. None of these risk factors or other patient characteristics (age, sex, tumour location and tumour stage) were correlated with the protein level of the individual CYP enzymes as determined by quantitation of immunoblot staining. However, the small series of samples precludes any strong conclusion concerning the lack of such correlations. There were no differences between squamous cell carcinomas and adenocarcinomas in either the qualitative or quantitative expression of the CYP enzymes. These data demonstrate that a range of CYP enzymes are expressed in human oesophageal mucosa and indicate that this tissue has the capacity to activate chemical carcinogens to reactive DNA binding metabolites.


Factors affecting exercise attendance and completion in sedentary older adults: a meta-analytic approach.
Many different constructs are used currently in the literature to assess exercise adherence. This study examined whether the same or different variables predict exercise attendance and exercise completion among sedentary older adults. Thirty-seven randomized control trials were selected from articles published between 1980 and 2000 that tested exercise interventions for sedentary older adults. Block-entry, weighted, hierarchical meta-regression analyses were conducted. Different factors predicted attendance and completion. Group-based (P < .05) and resistance exercise (P < .1) predicted higher attendance rates than individual-based and aerobic exercise. In contrast, facility-based exercise was associated with higher completion rates than homebased exercise (P < .1). Results show that completing a program is not synonymous with good attendance. Program designers need to consider different strategies to boost both of these rates that need to be maximized to best benefit program participants.


Large heterogeneity of mitochondrial DNA transcription and initiation of replication exposed by single-cell imaging.
Mitochondrial DNA (mtDNA) replication and transcription are crucial for cell function, but these processes are poorly understood at the single-cell level. We describe a novel fluorescence in situ hybridization protocol, called mTRIP (mitochondrial transcription and replication imaging protocol), that reveals simultaneously mtDNA and RNA, and that can also be coupled to immunofluorescence for in situ protein examination. mTRIP reveals mitochondrial structures engaged in initiation of DNA replication by identification of a specific sequence in the regulatory D-loop, as well as unique transcription profiles in single human cells. We observe and quantify at least three classes of mitochondrial structures: (i) replication initiation active and transcript-positive (Ia-Tp); (ii) replication initiation silent and transcript-positive (Is-Tp); and (iii) replication initiation silent and transcript-negative (Is-Tn). Thus, individual mitochondria are dramatically heterogeneous within the same cell. Moreover, mTRIP exposes a mosaic of distinct nucleic acid patterns in the D-loop, including H-strand versus L-strand transcripts, and uncoupled rRNA transcription and mtDNA initiation of replication, which might have functional consequences in the regulation of the mtDNA. Finally, mTRIP identifies altered mtDNA processing in cells with unbalanced mtDNA content and function, including in human mitochondrial disorders. Thus, mTRIP reveals qualitative and quantitative alterations that provide additional tools for elucidating the dynamics of mtDNA processing in single cells and mitochondrial dysfunction in diseases.


Efficient β-poly(l-malic acid) production from Jerusalem artichoke by Aureobasidium pullulans ipe-1 immobilized in luffa sponge matrices.
β-poly(l-malic acid) (PMLA) production by Aureobasidium pullulans ipe-1 using Jerusalem artichoke tuber (JA) hydrolysate as a low cost carbon source was developed. The PMLA production was favored by JA pretreated with 0.06 M nitric acid without adding exogenous nitrogen sources into fermentation medium. With an initial 130 g/L total sugar of the JA hydrolysate, the highest PMLA productivity 0.52 g/L·h was achieved, which was increased by 2.0 folds compared to that with sole glucose case. To further enhance PMLA productivity, the cells were immobilized in luffa sponge matrices, and repeated batch culture was carried out for 4 cycles. The resulting PMLA productivity was further enhanced by 50% compared with the batch culture. The cost of PMLA production in the JA case was only 5.4% of that in the glucose case. The outcomes of this work provided a strategy of PMLA production on a commercial scale.


Comprehensive Neuropsychiatric and Cognitive Characterization of Former Professional Football Players: Implications for Neurorehabilitation.
Objectives: To identify novel targets for neurorehabilitation of people with a remote history of multiple concussions by: (1) comprehensively characterizing neuropsychiatric and cognitive functioning in former professional football players, with a focus on executive functions; (2) distinguishing concussion-related findings from pre-morbid/cohort characteristics of professional football players; and, (3) exploring the relationship between executive functions and neuropsychiatric symptoms. Participants: Sixty-one high-functioning former professional football players and 31 age- and sex-matched control participants without history of concussion or participation in contact sports. Design: Between-groups analyses. Main measures: Neuropsychiatric. Personality Assessment Inventory (PAI) clinical scales plus the Aggression treatment consideration scale; the Mini International Neuropsychiatric Interview (MINI). Cognitive. Comprehensive clinical neuropsychological battery assessing domains of verbal and visuospatial attention; speed of processing and memory; current and estimated pre-morbid IQ; and, executive functioning, including two experimental measures that were novel for this population (i.e., response inhibition and inconsistency of responding on a go/no-go task). Results: (1) Compared to control participants, former professional football players scored significantly higher on the PAI Depression, Mania, and Aggression scales, and significantly lower on response inhibition. (2) Relative to controls, former players with >3 concussions ( x ¯ = 6 . 1 ), but not former players with ≤ 3 concussions ( x ¯ = 2 . 0 ), showed (i) significantly higher scores on the PAI Depression scale, (ii) significantly more MINI clinical diagnoses overall, and manic/hypomanic episodes specifically, and (iii) significantly poorer executive function. (3) Mediation analysis revealed that concussion exposure had a significant indirect effect on PAI Depression, Mania, and Aggression via inconsistency of responding on the go/no-go task. Conclusions: Notable impairments to neuropsychiatric functioning and worse performance on a sensitive experimental measure of executive function were observed; these were related to both concussion history and pre-morbid (cohort) factors. Therefore, neuropsychiatric and executive functioning should be carefully assessed in those with a remote history of multiple concussions. Moreover, former players' neuropsychiatric symptoms were associated with inconsistency of responding; this suggests that treatments targeted at response inconsistency could help to mitigate neuropsychiatric dysfunction.


Effects of coenzyme Q10 and α-lipoic acid supplementation in fructose fed rats.
This study was conducted to investigate the effects of α-lipoic acid and coenzyme Q10 on plasma levels of lipids, asymmetric dimethylarginine, oxidative stress in fructose fed rats which provide a model of dietary-induced insulin resistance and to evaluate vascular changes developing in these rats by histologically. Male Sprague Dawley rats were used in this study. The animals were divided into 4 groups. Group 1 did not receive any medication and served as a control. Group 2 received a regular diet and water ad libitum and fructose was administered as % 10 solution in drinking water. Group 3 received α-lipoic acid (100 mg/kg/day) i.p. for 5 weeks and Group 4 received coenzyme Q10 (10 mg/kg/day) i.p. for 5 weeks. For determination of plasma asymmetric dimethylarginine, glutathione and malondialdehyde levels, high-performance liquid chromatography system was used. Homeostatic model assessment as a measure of insulin resistance was calculated. Lipid profile measurements were determined using enzymatic assay on an Auto analyzer. The high fructose diet was significantly associated with an increase in levels of plasma LDL, VLDL and total cholesterol and decrease in level of HDL cholesterol. Plasma asymmetric dimethylarginine, malondialdehyde and glutathione levels were also increase in these rats. α-lipoic acid or coenzyme Q10 supplementation was found to have some positive effect on these parameters. These findings were also demonstrated by morphological observation of the aorta. We demonstrated that administration of α-lipoic acid and coenzyme Q10 notably suppresses oxidative and nitrative stress, hyperinsulinemia, insulin resistance developing in fructose fed rats, a model of metabolic syndrome (MS). These positive effects of α-lipoic acid or coenzyme Q10 can be attributed to its antioxidant activity.


Anaphylaxis to Lumason Contrast Agent in Echocardiography after Immune Sensitization: A Case Report and Literature Review.
With the continued evolution of echocardiography techniques, novel contrast agents have been introduced to aid in evaluating cardiac structure and function. Lumason is an FDA-approved ultrasound-enhancing agent made of Sulfur hexafluoride lipid-type A microspheres. We report a case of Lumason-induced immune sensitization and subsequent anaphylaxis on re-exposure in a middle-aged male patient presenting with symptoms concerning for a cerebrovascular accident. The patient was successfully treated with steroids and antihistamines. Providers, including physicians, echocardiography technicians, and mid-level providers should be aware of and be prepared to respond to the possibility of anaphylaxis to contrast-enhancing agents used in echocardiography.


Isolation of Streptococcus pneumoniae biofilm mutants and their characterization during nasopharyngeal colonization.
Asymptomatic colonization of the nasopharynx by Streptococcus pneumoniae precedes pneumococcal disease, yet pneumococcal colonization factors remain poorly understood. Many bacterial infections involve biofilms which protect bacteria from host defenses and antibiotics. To gain insight into the genetics of biofilm formation by S. pneumoniae, we conducted an in vitro screen for biofilm-altered mutants with the serotype 4 clinical isolate TIGR4. In a first screen of 6,000 mariner transposon mutants, we repeatedly isolated biofilm-overproducing acapsular mutants, suggesting that the capsule was antagonistic to biofilm formation. Therefore, we screened 6,500 additional transposon mutants in an S. pneumoniae acapsular background. Following this approach, we isolated 69 insertions in 49 different genes. The collection of mutants includes genes encoding bona fide and putative choline binding proteins, adhesins, synthases of membrane and cell wall components, extracellular and cell wall proteases, efflux pumps, ABC and PTS transporters, and transcriptional regulators, as well as several conserved and novel hypothetical proteins. Interestingly, while four insertions mapped to rrgA, encoding a subunit of a recently described surface pilus, rrgB and rrgC (encoding the other two pilus subunits) mutants had no biofilm defects, implicating the RrgA adhesin but not the pilus structure per se in biofilm formation. To correlate our findings to the process of colonization, we transferred a set of 29 mutations into the wild-type encapsulated strain and then tested the fitness of the mutants in vivo. Strikingly, we found that 23 of these mutants were impaired for nasopharyngeal colonization, thus establishing a link between biofilm formation and colonization.


Patients treated for psychosis and their perceptions of care in compulsory treatment: basis for an action plan.
The purpose of the study was to describe patients' conceptions and experiences of care in compulsory treatment for acute onset of psychosis. Twelve patients with experience of compulsory treatment were interviewed in 2008-2009, and phenomenographic analysis was used to analyse the data. Two descriptive categories emerged in the results: receiving needed support and perceiving respectful care. Patients perceived that coercive interventions were positive if they were given good care, if they were given the shelter they needed, if they got help with understanding and if the setting was healing. Patients felt respected if they were treated like human beings, if they were allowed to retain as much of their autonomy as possible and if they were invited to participate even though they were under compulsory treatment. The results show that it is important to prevent patients from being traumatized during compulsory treatment and to take advantage of patients' inner resources. Patients' experiences of compulsory treatment can form the basis for preparing an individual action plan for future compulsory treatment. Individual action plans could empower patients during compulsory treatment and improve their experience of care.


Pulsed UV-C disinfection of Escherichia coli with light-emitting diodes, emitted at various repetition rates and duty cycles.
A 2010 study exposed Staphylococcus aureus to ultraviolet (UV) radiation and thermal heating from pulsed xenon flash lamps. The results suggested that disinfection could be caused not only by photochemical changes from UV radiation, but also by photophysical stress damage caused by the disturbance from incoming pulses. The study called for more research in this area. The recent advances in light-emitting diode (LED) technology include the development of LEDs that emit in narrow bands in the ultraviolet-C (UV-C) range (100-280 nm), which is highly effective for UV disinfection of organisms. Further, LEDs would use less power, and allow more flexibility than other sources of UV energy in that the user may select various pulse repetition frequencies (PRFs), pulse irradiances, pulse widths, duty cycles and types of waveform output (e.g. square waves, sine waves, triangular waves, etc.). Our study exposed Escherichia coli samples to square pulses of 272 nm radiation at various PRFs and duty cycles. A statistically significant correlation was found between E. coli's disinfection sensitivity and these parameters. Although our sample size was small, these results show promise and are worthy of further investigation. Comparisons are also made with pulsed disinfection by LEDs emitting at 365 nm, and pulsed disinfection by xenon flash lamps.


Determining the Most Appropriate Assistive Walking Device Using the Inertial Measurement Unit-Based Gait Analysis System in Disabled Patients.
To evaluate the gait pattern of patients with gait disturbances without consideration of defilades due to assistive devices. This study focuses on gait analysis using the inertial measurement unit (IMU) system, which can also be used to determine the most appropriate assistive device for patients with gait disturbances. Records of 18 disabled patients who visited the Department of Rehabilitation from May 2018 to June 2018 were selected. Patients' gait patterns were analyzed using the IMU system with different assistive devices to determine the most appropriate device depending on the patient's condition. Evaluation was performed using two or more devices, and the appropriate device was selected by comparing the 14 parameters of gait evaluation. The device showing measurements nearer or the nearest to the normative value was selected for rehabilitation. The result of the gait evaluation in all 18 patients was analyzed using the IMU system. According to the records, the patients were evaluated using various assistive devices without consideration of defilades. Moreover, this gait analysis was effective in determining the most appropriate device for each patient. Increased gait cycle time and swing phase and decreased stance phase were observed in devices requiring significant assistance. The IMU-based gait analysis system is beneficial in evaluating gait in clinical fields. Specifically, it is useful in evaluating patients with gait disturbances who require assistive devices. Furthermore, it allows the establishment of an evidence-based decision for the most appropriate assistive walking devices for patients with gait disturbances.


Thrombolysis for myocardial infarction.
Treating myocardial infarction by dissolving occlusive thrombi in coronary arteries is an attractive idea. Although some thrombolytic agents have been available for many years their use in this role has only recently been critically examined. The place of thrombolysis in the management of myocardial infarction has yet to be determined.


Caveolin 3 is associated with the calcium release complex and is modified via in vivo triadin modification.
The triadin isoforms Trisk 95 and Trisk 51 are both components of the skeletal muscle calcium release complex. To investigate the specific role of Trisk 95 and Trisk 51 isoforms in muscle physiology, we overexpressed Trisk 95 or Trisk 51 using adenovirus-mediated gene transfer in skeletal muscle of newborn mice. Overexpression of either Trisk 95 or Trisk 51 alters the muscle fiber morphology, while leaving unchanged the expression of the ryanodine receptor, the dihydropyridine receptor, and calsequestrin. We also observe an aberrant expression of caveolin 3 in both Trisk 95- and Trisk 51-overexpressing skeletal muscles. Using a biochemical approach, we demonstrate that caveolin 3 is associated with the calcium release complex in skeletal muscle. Taking advantage of muscle and non-muscle cell culture models and triadin null mouse skeletal muscle, we further dissect the molecular organization of the caveolin 3-containing calcium release complex. Our data demonstrate that the association of caveolin 3 with the calcium release complex occurs via a direct interaction with the transmembrane domain of the ryanodine receptor. Taken together, these data suggest that caveolin 3-containing membrane domains and the calcium release complex are functionally linked and that Trisk 95 and Trisk 51 are instrumental to the regulation of this interaction, the integrity of which may be crucial for muscle physiology.


Do people really say it is "likely" when they believe it is only "possible"? Effect of politeness on risk communication.
Politeness theory posits that speakers can use verbal probabilities (e.g., there is a chance, it is likely) to hedge bad news. So far, only indirect evidence supports that claim: From the hearer's standpoint, verbal probabilities are interpreted either as plain likelihood-communication devices or as face-management devices, resulting in different risk perceptions. The present research aims to test more directly the postulate of politeness theory by focusing on the effects of speakers' intentions on risk communication. In three experiments, participants communicated a probability by choosing an expression from a list of verbal probabilities. Results consistently showed that polite speakers communicated a different risk magnitude than informative speakers. Further findings indicate that the effect of the speakers' intention depends on the valence of the uncertain outcome. The theoretical and applied implications of these findings are discussed.


The role of CcpA transcriptional regulator in carbon metabolism in Bacillus subtilis.
The CcpA protein has been identified as a key regulator of carbon metabolism in Bacillus subtilis. CcpA is a DNA binding protein in the LacI/GalR transcriptional repressor family, and genes which respond to CcpA contain common cis-acting target sequences (Ccp boxes). A number of pathways involved in carbon source utilization are repressed by CcpA, while at least one gene which is involved in excretion of excess carbon is activated by CcpA. Genes repressed by CcpA generally contain Ccp boxes within or downstream of the promoter, while ackA, which is activated by CcpA, contains Ccp boxes upstream of the promoter. It therefore appears that CcpA acts globally to direct carbon flow in B. subtilis.


Specific immunity in rats inoculated with Trypanosoma brucei brucei derived from tsetse salivary glands and cultured in vitro.
Twenty rats were each immunized intraperitoneally with 1.25 X 10(7) formalin-fixed Trypanosoma brucei brucei (Stock EATRO 1969) derived from tsetse salivary glands and propagated in culture at 29 degrees C in their infective form for more than 1000 days. Another group, consisting of 10 rats, was similarly immunized with parasites of stock EATRO 999 derived from salivary glands of a tsetse fly and propagated in culture for more than 360 days. Immunized animals were resistant to homologous challenge with 10(4) cultured infective parasites. There was no cross protection between stocks and animals were susceptible to homologous infective tsetse bite challenge.


Multicarbohydrase Enzymes for Non-ruminants.
The first purpose of this review is to outline some of the background information necessary to understand the mechanisms of action of fibre-degrading enzymes in non-ruminants. Secondly, the well-known and understood mechanisms are described, i) eliminating the nutrient encapsulating effect of the cell wall and ii) ameliorating viscosity problems associated with certain Non Starch Polysaccharides, particularly arabinoxylans and β-glucans. A third, indirect mechanism is then discussed: the activity of such enzymes in producing prebiotic oligosaccharides and promoting beneficial cecal fermentation. The literature contains a wealth of information on various non starch polysaccharide degrading enzyme (NSPase) preparations and this review aims to conclude by discussing this body of work, with reference to the above mechanisms. It is suggested that the way in which multi- versus single-component products are compared is often flawed and that some continuity should be employed in methods and terminology.


Effect of DNA repair inhibitor AsiDNA on the incidence of telomere fusion in crisis.
Telomere fusions lead to a state of genomic instability, and are thought to drive clonal evolution and tumorigenesis. Telomere fusions occur via both Classical and Alternative Non-Homologous End Joining repair pathways. AsiDNA is a DNA repair inhibitor that acts by mimicking a DNA double strand break (DSB) and hijacking the recruitment of proteins involved in various DNA repair pathways. In this study, we investigated whether the inhibition of DSB-repair pathways by AsiDNA could prevent telomere fusions during crisis. The present study showed that AsiDNA decreased the frequency of telomere fusions without affecting the rate of telomere erosion. Further, it indicated that AsiDNA does not impact the choice of the repair pathway used for the fusion of short dysfunctional telomeres. AsiDNA is thought to prevent short telomeres from fusing by inhibiting DNA repair. An alternative, non-mutually exclusive possibility is that cells harbouring fusions preferentially die in the presence of AsiDNA, thus resulting in a reduction in fusion frequency. This important work could open the way for investigating the use of AsiDNA in the treatment of tumours that have short dysfunctional telomeres and/or are experiencing genomic instability.


Targeted therapy with a localised abluminal groove, low-dose sirolimus-eluting, biodegradable-polymer coronary stent - five-year results of the TARGET All Comers randomised clinical trial.
In the prospective, multicentre, randomised TARGET All Comers study, percutaneous coronary intervention (PCI) with the FIREHAWK biodegradable-polymer sirolimus-eluting stent (BP-SES) was non-inferior to the durable-polymer everolimus-eluting stent (DP-EES) for the primary endpoint of target lesion failure (TLF) at 12 months. We aimed to report the final study outcomes at 5 years. Patients referred for PCI were randomised to receive either a BP-SES or DP-EES in a 1:1 ratio in 10 European countries. Randomisation was stratified by centre and ST-elevation myocardial infarction (STEMI) presentation, and clinical follow-up extended to 5 years. The primary endpoint was TLF (composite of cardiac death, target vessel myocardial infarction [MI], or ischaemia-driven target lesion revascularisation). Secondary endpoints included patient-oriented composite events (POCE; composite of all-cause death, all MI, or any revascularisation and its components). From December 2015 to October 2016, 1,653 patients were randomly assigned to the BP-SES or DP-EES groups, of which 93.8% completed 5-year clinical follow-up or were deceased. At 5 years, TLF occurred in 17.1% of the BP-SES group and in 16.3% of the DP-EES group (p=0.68). POCE occurred in 34.0% of the BP-SES group and 32.7% of the DP-EES group (p=0.58). Revascularisation was the most common POCE, occurring in 19.3% of patients receiving BP-SES and 19.2% receiving DP-EES, of which less than one-third was ischaemia-driven target lesion-related. In the landmark analysis, there were no differences in the rates of TLF and POCE between groups from 1 to 5 years, and these results were consistent across all subgroups. In an all-comers population requiring stent implantation for myocardial ischaemia, the BP-SES was non-inferior to the DP-EES for the primary endpoint of TLF at 12 months, and results were sustained at 5 years, confirming the long-term safety and efficacy of the FIREHAWK BP-SES.


Exocytosis in mast cells by basic secretagogues: evidence for direct activation of GTP-binding proteins.
Histamine release induced by the introduction of a nonhydrolyzable analogue of GTP, GTP-gamma-S, into ATP-permeabilized mast cells, is associated with phosphoinositide breakdown, as evidenced by the production of phosphatidic acid (PA) in a neomycin-sensitive process. The dependency of both PA formation and histamine secretion on GTP-gamma-S concentrations is bell shaped. Whereas concentrations of up to 0.1 mM GTP-gamma-S stimulate both processes, at higher concentrations the cells' responsiveness is inhibited. At a concentration of 1 mM, GTP-gamma-S self-inhibits both PA formation and histamine secretion. Inhibition of secretion can, however, be overcome by the basic secretagogues compound 48/80 and mastoparan that in suboptimal doses synergize with 1 mM GTP-gamma-S to potentiate secretion. Secretion under these conditions is not accompanied by PA formation and is resistant both to depletion of Ca2+ from internal stores and to pertussis toxin (PtX) treatment. In addition, 48/80, like mastoparan, is capable of directly stimulating the GTPase activity of G-proteins in a cell-free system. Together, our results are consistent with a model in which the continuous activation of a phosphoinositide-hydrolyzing phospholipase C (PLC) by a stimulatory G-protein suffices to trigger histamine secretion. Basic secretagogues of mast cells, such as compound 48/80 and mastoparan, are capable of inducing secretion in a mechanism that bypasses PLC by directly activating a G-protein that is presumably located downstream from PLC (GE). Thereby, these secretagogues induce histamine secretion in a receptor-independent manner.


Could the [14C]urea breath test be proposed as a 'gold standard' for detection of Helicobacter pylori infection ?
The urea breath test (UBT) with a microdose of [14C] is a non-invasive and simple method for the assessment of gastric infection by Helicobacter pylori. The aim of this study was to compare the [14C]UBT with invasive methods widely used for assessment of H. pylori gastric infection, including histology with hematoxylin-eosin staining, the gastric smear technique using Giemsa staining, and the biopsy urease test. We evaluated patients referred to our clinic for elective upper gastrointestinal endoscopy excluding those on antibiotics and/or bismuth during the previous 4 weeks, patients on H+ blockers or H2 antihistamines during the previous 7 days, pregnant women, and patients who had undergone gastric surgery or had bleeding disorders. Eighty-nine patients ranging in age from 18-75 years were included in the final study population, 61 women and 28 men (mean age: 43(15 years). When histology alone was considered as the reference standard, sensitivity for the [14 C]UBT was 94%, with a specificity of 37%; when the Giemsa technique, sensitivity was 95%, and specificity 35%; and when the biopsy urease test, sensitivity was 94% and specificity 45%. With two or more invasive techniques together considered as the reference standard, the [14C]UBT had a sensitivity of 95%, with a specificity of 44%. [14C]UBT is an objective and reproducible technique, capable of sampling the whole gastric mucosa. It shows high sensitivity, but low specificity, which could be explained by limited gastric sampling plus subjective interpretation in the invasive techniques that are currently used as gold standard.


Protein and Small-Molecule Leucopoiesis and Thrombopoiesis Stimulators.
Pluripotent stem cells of the bone marrow are stimulated by different cytokines to proliferation and differentiation into various types of blood cells. These cytokines are mostly glycoproteins. Erythropoietin stimulates stem cells to the formation of erythrocytes while colony-stimulating factors cause the formation of different types of white blood cells. Stem cell factors play an important role in the maintenance and survival of blood cells of all types. Thrombopoietin stimulates stem cells to proliferation and formation of blood platelets. Granulocyte colony-stimulating factor is probably the most important drug in use. It stimulates stem cells to the formation of neutrophile granulocytes. It is often used in recombinant forms such as filgrastim in the treatment of neutropenia in cancer chemotherapy or AIDS. Its pegylated conjugates such as pegfilgrastim are also available. Its activity can be supported by plerixafor, a small molecule - bicyclam derivative acting as an indirect agonist of stem cells factor. It acts as an antagonist of CXCR4 receptor activation of which brakes hematopoiesis. The treatment of conditions accompanied by thrombocytopenia such as idiopathic thrombocytopenic purpura is currently not performed by thrombopoietin but synthetic agonists of its receptor are preferred. Romiplostim is a peptibody. It consists of a protein part interacting with the thrombopoietin receptor which is, however, different from thrombopoietin, and of Fc fragment of immunoglobulin G1. In contrast, small molecule thrombopoietin receptor agonists represented by eltrombopag can be given orally unlike all of the above.


[Activity of hippocampal neurons deprived of ascending brain stem connections].
97 units of the hippocampal fields CA3 and CA1 were recorded extracellularly in chronic unanaesthetized rabbits after complete basal undercutting of the septum. In activity of about one third of the units slow rhythmic bursts (3,3-4,5 Hz) were present. Low frequency theta-like rhythm was present in EEG of the hippocampus. Reactivity to sensory stimuli was unusually low (46-47% of reactive units). Specific and phasic effects of stimuli, typical of the normal field CA1, were observed in both fields. The majority of the reactive neurons respond to sensory stimuli by prolonged shift of the mean frequency of discharges, by regularization of the rhythmic component, or by gradual increase of diffuse activation. Effects of suppression of activity by sensory stimuli were very rare. The data are discussed in the light of presumed difference of reinnervation by axonal sprouting in conditions of basal undercutting of the septum and complete septo-hippocampal disconnection.


The long-term effect of vagus nerve stimulation on quality of life in patients with pharmacoresistant focal epilepsy: the PuLsE (Open Prospective Randomized Long-term Effectiveness) trial.
To evaluate whether vagus nerve stimulation (VNS) as adjunct to best medical practice (VNS + BMP) is superior to BMP alone in improving long-term health-related quality of life (HRQoL). PuLsE (Open Prospective Randomized Long-term Effectiveness) was a prospective, randomized, parallel-group, open-label, and long-term effectiveness study (conducted at 28 sites in Europe and Canada). Adults with pharmacoresistant focal seizures (n = 112) received VNS + BMP or BMP (1:1 ratio). Medications and VNS parameters could be adjusted as clinically indicated for optimal seizure control while minimizing adverse effects. Primary endpoint was mean change from baseline HRQoL (using Quality of Life in Epilepsy Inventory-89 total score; QOLIE-89). Secondary endpoints included changes in seizure frequency, responder rate (≥50% decrease in seizure frequency), Centre for Epidemiologic Studies Depression scale (CES-D), Neurological Disorders Depression Inventory-Epilepsy scale (NDDI-E), Clinical Global Impression-Improvement scale (CGI-I), Adverse Event Profile (AEP), and antiepileptic drug (AED) load. The study was prematurely terminated due to recruitment difficulties prior to completing the planned enrollment of n = 362. Results for n = 96 who had baseline and at least one follow-up QOLIE-89 assessment (from months 3-12) were included in this analysis. Mixed model repeated measures (MMRM) analysis of variance was performed on change from baseline for the primary and secondary endpoints. Significant between-group differences in favor of VNS + BMP were observed regarding improvement in HRQoL, seizure frequency, and CGI-I score (respective p-values < 0.05, 0.03, and 0.01). More patients in the VNS + BMP group (43%) reported adverse events (AEs) versus BMP group (21%) (p = 0.01), a difference reflecting primarily mostly transient AEs related to VNS implantation or stimulation. No significant difference between treatment groups was observed for changes in CES-D, NDDI-E, AEP, and AED load. VNS therapy as a treatment adjunct to BMP in patients with pharmacoresistant focal seizures was associated with a significant improvement in HRQoL compared with BMP alone. A PowerPoint slide summarizing this article is available for download in the Supporting Information section here.


Evaluation of Maxillofacial Characteristics in Individuals with Anterior Open Bite Using Denture Frame Analysis.
The aim of this study was to evaluate the effectiveness of denture frame analysis (DFA) in individuals with anterior open bite who had completed pubertal peak growth (post-peak). This retrospective study was conducted using the cephalometric radiographs and hand-wrist X-rays of 50 individuals with open bite (mean age: 17.33±3 years; 35 female, 15 male) and a control group of 50 individuals without open bite (mean age: 17.38±2.72; 35 female, 15 male). All individuals included in the study were skeletal Class I and had completed or nearly completed skeletal growth. Skeletal and dental measurements pertaining to DFA were done and the data were analyzed using independent samples t test and Mann-Whitney U test. Measurements assessing the vertical dimension showed that GoGn/SN, Frankfurt horizontal (FH)/mandibular plane (MP), palatal plane (PP)/MP, occlusal plane (OP)/MP, and OP-MP/PP-MP measurements were significantly greater in the open bite group (p<0.001). AB/MP angle was significantly larger in the control group (p<0.05). The open bite group had shorter A'-P' (posterior maxillary length; p<0.05) and, therefore, higher A'-6'/A'-P' ratio (p<0.01). Our results suggest that there may be a close association between maxillary OP inclination and mandibular position in individuals with open bite, and that open bite may arise due to maxillary denture base deficiency, especially in the posterior region. DFA may be useful in the differential diagnosis of open bite and in treatment planning, particularly when determining the need for tooth extraction.


A novel site contributing to growth-arrest-specific gene 6 binding to its receptors as revealed by a human monoclonal antibody.
Gas6 (growth-arrest-specific gene 6) is a vitamin K-dependent protein known to activate the Axl family of receptor tyrosine kinases. It is an important regulator of thrombosis and many other biological functions. The C-terminus of Gas6 binds to receptors and consists of two laminin-like globular domains LG1 and LG2. It has been reported that a Ca2+-binding site at the junction of LG1 and LG2 domains and a hydrophobic patch at the LG2 domain are important for receptor binding [Sasaki, Knyazev, Cheburkin, Gohring, Tisi, Ullrich, Timpl and Hohenester (2002) J. Biol. Chem. 277, 44164-44170]. In the present study, we developed a neutralizing human monoclonal antibody, named CNTO300, for Gas6. The antibody was generated by immunization of human IgG-expressing transgenic mice with recombinant human Gas6 protein and the anti-Gas6 IgG sequences were rescued from an unstable hybridoma clone. Binding of Gas6 to its receptors was partially inhibited by the CNTO300 antibody in a dose-dependent manner. To characterize further the interaction between Gas6 and this antibody, the binding kinetics of CNTO300 for recombinant Gas6 were compared with independently expressed LG1 and LG2. The CNTO300 antibody showed comparable binding affinity, yet different dependence on Ca2+, to Gas6 and LG1. No binding to LG2 was detected. In the presence of EDTA, binding of the antibody to Gas6 was disrupted, but no significant effect of EDTA on LG1 binding was evident. Further epitope mapping identified a Gas6 peptide sequence recognized by the CNTO300 antibody. This peptide sequence was found to be located at the LG1 domain distant from the Ca2+-binding site and the hydrophobic patch. Co-interaction of Gas6 with its receptor and CNTO300 antibody was detected by BIAcore analysis, suggesting a second receptor-binding site on the LG1 domain. This hypothesis was further supported by direct binding of Gas6 receptors to an independently expressed LG1 domain. Our results revealed, for the first time, a second binding site for Gas6-receptor interaction.


Isokinetic concentric quadriceps and hamstring normative data for elite collegiate American football players participating in the NFL Scouting Combine.
Isokinetic concentric quadriceps and hamstring strength data using a Cybex dynamometer are collected for elite collegiate American football players invited to the annual National Football League Scouting Combine. We constructed a normative (reference) database of the Cybex strength data for the purpose of allowing comparison of an individual's values to his peers. Data reduction was performed to construct frequency distributions of hamstring/quadriceps (H/Q) ratios and side-to-side strength differences. For the cohort (n = 1,252 players), a statistically significant but very small (1.9%) mean quadriceps strength preference existed for dominant side vs. nondominant side. Peak torque (Newton meters, best repetition) for quadriceps and hamstrings was significantly correlated to player body mass (weight) (the same relationship was found for other variables using peak torque in the calculation). Peak torque varied by player position, being greatest for offensive linemen and lowest for kickers (p < 0.0001). Adjusting for body weight overcorrected these differences. The H/Q ratios and frequency distributions were similar across positions, with a mean of 0.6837 ± 0.137 for the cohort dominant side vs. 0.6940 ± 0.145 for the nondominant side (p = 0.021, n = 1,252). Considerable variation was seen for dominant-to-nondominant side difference for peak torque. For quadriceps, 47.2% of players had differences between -10% and +10%, 21.0% had a peak torque dominant-side deficit of 10% or greater compared to nondominant side, and for 31.8% of players, dominant-side peak torque was greater than 10% compared to nondominant side. For hamstrings, 57.0% of players had differences between -10% and +10%, 19.6% had a peak torque dominant-side deficit of 10% or greater compared to nondominant side, and 23.4% of players, dominant-side peak torque was greater than 10% compared to nondominant side. We observed that isokinetic absolute strength variables are dependent on body weight and vary across player position. The H/Q ratios vary only within a relatively narrow range. Side-to-side differences in strength variables >10% are common, not the exception.


One-to-one registration of en-face optical coherence tomography attenuation coefficients with histology of a prostatectomy specimen.
Optical coherence tomography (OCT), enables high-resolution 3D imaging of the morphology of light scattering tissues. From the OCT signal, parameters can be extracted and related to tissue structures. One of the quantitative parameters is the attenuation coefficient; the rate at which the intensity of detected light decays in depth. To couple the quantitative parameters with the histology one-to-one registration is needed. The primary aim of this study is to validate a registration method of quantitative OCT parameters to histological tissue outcome through one-to-one registration of OCT with histology. We matched OCT images of unstained fixated prostate tissue slices with corresponding histology slides, wherein different histologic types were demarcated. Attenuation coefficients were determined by a supervised automated exponential fit (corrected for point spread function and sensitivity roll-off related signal losses) over a depth of 0.32 mm starting from 0.10 mm below the automatically detected tissue edge. Finally, the attenuation coefficients corresponding to the different tissue types of the prostate were compared. From the attenuation coefficients, we produced the squared relative residue and goodness-of-fit metric R2 . This article explains the method to perform supervised automated quantitative analysis of OCT data, and the one-to-one registration of OCT extracted quantitative data with histopathological outcomes.


Cigarette smoking and cancer: intensity patterns in the alpha-tocopherol, beta-carotene cancer prevention study in Finnish men.
Relative risks for lung and bladder cancers by smoking intensity level off at more than 15-20 cigarettes per day. A three-parameter excess relative risk model in pack-years and intensity quantified this leveling (Lubin et al., Am J Epidemiol 2007;166:479-89). Above 15-20 cigarettes per day was an "inverse exposure rate" effect whereby, for equal pack-years, the excess relative risk/pack-year decreased with increasing intensity; that is, smoking at a lower intensity for a longer duration was more deleterious than smoking at a higher intensity for a shorter duration. After adjustment for pack-years, intensity effects were quantitatively homogeneous across multiple case-control studies of lung, bladder, oral cavity, pancreas, and esophagus cancers. The authors extended those analyses to examine intensity patterns for incident bladder, esophagus, kidney, larynx, liver, lung, oropharynx, and pancreas cancers by using data from a single prospective cohort in Finland, the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, with follow-up from enrollment, which occurred between 1985 and 1988, through April 2004. At more than 10 cigarettes per day, they found an inverse exposure rate pattern for each cancer site. After adjustment for pack-years, intensity effects were quantitatively homogeneous across the diverse cancer sites and homogeneous with intensity effects from the prior analysis of multiple studies. Consistency of intensity patterns suggested a general phenomenon and may provide clues to the molecular basis of smoking-related cancer risk.


Inhibitory Components from Glycosmis stenocarpa on Pepper Mild Mottle Virus.
The goal of this study was to identify a source of natural plant compounds with inhibitory activity against pepper mild mottle virus (PMMoV). We showed, using a half-leaf assay, that murrayafoline-A (1) and isomahanine (2) isolated from the aerial parts of Glycosmis stenocarpa have inhibitory activity against PMMoV through curative, inactivation, and protection effects. Using a leaf-disk assay, we confirmed that 2 inhibited virus replication in Nicotiana benthamiana. Using electron microscopy, we found that a mixture of the virus with 2 resulted in damage to the rod-shaped virus.


Functions of two genes in aluminium (Al) stress resistance: repression of oxidative damage by the AtBCB gene and promotion of efflux of Al ions by the NtGDI1gene.
The functions of two genes whose expression provides tolerance to aluminium (Al) stress were investigated using plants and Saccharomyces cerevisiae (yeast): the Arabidopsis thaliana blue copper binding gene (AtBCB) and Nicotiana tabacum guanosine diphosphate (GDP) dissociation inhibitor gene (NtGDI1). To determine the localization of these proteins, each gene was fused to the green fluorescent protein (GFP) gene and introduced into onion epidermal cells. AtBCB was localized to cell membrane region and NtGDI1 to cytoplasm. Transgenic lines over-expressing the AtBCB gene showed constitutive lignin production in whole roots. By contrast, wild-type Arabidopsis (Ler) produced a negligible level of lignin and enhanced lignin production in the root-tip region by Al stress. Compared with Ler, the AtBCB-expressing lines showed a lower deposition of malon dialdehyde after Al stress. Microscopic observation of the Al-treated roots indicated that the deposition of lipid peroxides was clearly low in the area where lignin accumulated. It was proposed that lipid peroxidation caused by Al stress was diminished by the formation of lignin. Expression of the NtGDI1 gene in yeast complemented the temperature-sensitive phenotype of a sec19 mutant at 37 degrees C. This gene also complemented an Al-sensitive phenotype shown by the sec19 mutant at the permissive temperature of 32 degrees C. These results suggested that the yeast Sec19 vesicle transport system has a function in providing basal Al resistance in yeast by the export of Al ions. It was also proposed that over-expression of the NtGDI1 protein activates an Al efflux system that protects Arabidopsis against Al toxicity.


Mitochondrial dysfunction in central nervous system white matter disorders.
Defects of mitochondrial respiration and function had been proposed as a major culprit in the most common neurodegenerative diseases, including prototypic diseases of central nervous system (CNS) white matter such as multiple sclerosis. The importance of mitochondria for white matter is best exemplified in a group of defects of the mitochondria oxidative metabolism called mitochondria leukoencephalopathies or encephalomyopathies. These diseases are clinically and genetically heterogeneous, given the dual control of the respiratory chain by nuclear and mitochondrial DNA, which makes the precise diagnosis and classification challenging. Our understanding of disease pathogenesis is nowadays still limited. Here, we review current knowledge on pathogenesis and genetics, outlining diagnostic clues for the various forms of mitochondria disease. In particular, we underscore the value of magnetic resonance imaging (MRI) for the differential diagnosis of specific types of mitochondrial leukoencephalopathies, such as genetic defects on SDHFA1. The use of novel technologies for gene identification, such as whole-exome sequencing studies, is expected to shed light on novel molecular etiologies, broadening prenatal diagnosis, disease understanding, and therapeutic options. Current treatments are mostly palliative, but very promising novel gene and pharmacologic therapies are emerging, which may also benefit a growing list of secondary mitochondriopathies, such as the peroxisomal disease adrenoleukodystrophy.


Progression of periodontal disease and interleukin-10 gene polymorphism.
Interleukin-10 is a key immunoregulatory cytokine that may be of significance in the immunopathogenesis of chronic inflammatory diseases such as periodontal disease. Molecular genetic studies have defined a number of haplotypes that may be associated with differing levels of interleukin-10 secretion. The present study investigated the possible association between interleukin-10 gene polymorphism and periodontal disease progression. Genomic DNA was obtained from 252 adults who were part of a prospective longitudinal study on the progression of periodontal disease in a general adult Australian population. Single nucleotide polymorphisms at positions -592 and -1082 in the interleukin-10 promoter were analysed using an induced heteroduplex methodology and used to determine interleukin-10 promoter haplotypes in individual samples. Periodontitis progression was assessed by measuring probing depths and relative attachment levels at regular intervals over a 5-year period. A generalized linear model was used to analyse the data, with age, gender, smoking status, interleukin-1 genotype and Porphyromonas gingivalis included as possible confounders. There was a significant (p approximately 0.02) main effect of interleukin-10 haplotypes, with individuals having either the ATA/ACC or the ACC/ACC genotype experiencing around 20% fewer probing depths of >or= 4 mm compared to individuals with other genotypes. Age and smoking had significant (p < 0.001) additional effects. These data suggest that the interleukin-10 genotype contributes to the progression of periodontal disease.


Vision-Based Assessment of Gait Features Associated With Falls in People With Dementia.
Gait impairments contribute to falls in people with dementia. In this study, we used a vision-based system to record episodes of walking over a 2-week period as participants moved naturally around their environment, and from these calculated spatiotemporal, stability, symmetry, and acceleration gait features. The aim of this study was to determine whether features of gait extracted from a vision-based system are associated with falls, and which of these features are most strongly associated with falling. Fifty-two people with dementia admitted to a specialized dementia unit participated in this study. Thirty different features describing baseline gait were extracted from Kinect recordings of natural gait over a 2-week period. Baseline clinical and demographic measures were collected, and falls were tracked throughout the participants' admission. A total of 1,744 gait episodes were recorded (mean 33.5 ± 23.0 per participant) over a 2-week baseline period. There were a total of 78 falls during the study period (range 0-10). In single variable analyses, the estimated lateral margin of stability, step width, and step time variability were significantly associated with the number of falls during admission. In a multivariate model controlling for clinical and demographic variables, the estimated lateral margin of stability (p = .01) was remained associated with number of falls. Information about gait can be extracted from vision-based recordings of natural walking. In particular, the lateral margin of stability, a measure of lateral gait stability, is an important marker of short-term falls risk.


Copy number variations and risk for schizophrenia in 22q11.2 deletion syndrome.
22q11.2 Deletion Syndrome (22q11.2DS) is a common microdeletion syndrome with congenital and late-onset features. Testing for the genomic content of copy number variations (CNVs) may help elucidate the 22q11.2 deletion mechanism and the variable clinical expression of the syndrome including the high (25%) risk for schizophrenia. We used genome-wide microarrays to assess CNV content and the parental origin of 22q11.2 deletions in a cohort of 100 adults with 22q11.2DS (44 with schizophrenia) and controls. 22q11.2DS subjects with schizophrenia failed to exhibit de novo CNVs or any excess of novel inherited CNVs outside the 22q11.2 region. There were no significant effects of parental origin of the 22q11.2 deletion, deletion length, parental age or family history on expression of schizophrenia. There was no evidence for a general increase of de novo CNVs in 22q11.2DS. A novel finding was the relative paucity of males with de novo 22q11.2 deletions of paternal origin (P = 0.019). The Y chromosome may play a mediating role in the mechanism of 22q11.2 deletion events during spermatogenesis, resulting in the previously observed excess of maternal de novo 22q11.2 deletions. Hemizygosity of the 22q11.2 region appears to be the major CNV-related risk factor for schizophrenia in 22q11.2DS. The results reinforce the need for further efforts to identify specific molecular mechanisms underlying this expression and to identify the 1% of patients with schizophrenia who carry 22q11.2 deletions.


Do 'do not attends' at a genitourinary medicine service matter?
Patients who do not attend (DNA) clinic appointments are wasteful of resources and may also pose a potential public health risk through the onward transmission of untreated sexually transmitted infections (STIs). This service evaluation was performed to assess the demographics of DNAs, patient reasons for non-attendance and follows up data to determine the health outcomes of non-attendance. The genitourinary (GU) medicine clinic DNA rate between October 2008 and March 2009 was 6% (383/6961). Of the 383 patients who DNA'd their appointment successful telephone contact was made in 182 (48%) of them. Of these 40% of patients reported that they had forgotten they held an appointment; no patients reported that clinic opening hours prevented their attendance. Telephone contact increased GU medicine clinic attendance by 9% (35/383), but led to a greater number of subsequent DNAs; 43/75 (63%) of patients who accepted a further appointment DNA'd that appointment. Sending a SMS text message to patients who DNA notifying them of the time of walk-in services might be the most effective way of recapturing these patients without compromising future clinic appointment slots.


External validation of three dimensional conformal radiotherapy based NTCP models for patient-rated xerostomia and sticky saliva among patients treated with intensity modulated radiotherapy.
The purpose of this study was to investigate the ability of predictive models for patient-rated xerostomia (XER(6M)) and sticky saliva (STIC(6M)) at 6 months after completion of primary (chemo)radiation developed in head and neck cancer patients treated with 3D-conformal radiotherapy (3D-CRT) to predict outcome in patients treated with intensity modulated radiotherapy (IMRT). Recently, we published the results of a prospective study on predictive models for patient-rated xerostomia and sticky saliva in head and neck cancer patients treated with 3D-CRT (3D-CRT based NTCP models). The 3D-CRT based model for XER(6M) consisted of three factors, including the mean parotid dose, age, and baseline xerostomia (none versus a bit). The 3D-CRT based model for STIC(6M) consisted of the mean submandibular dose, age, the mean sublingual dose, and baseline sticky saliva (none versus a bit). In the current study, a population consisting of 162 patients treated with IMRT was used to test the external validity of these 3D-CRT based models. External validity was described by the explained variation (R(2) Nagelkerke) and the Brier score. The discriminative abilities of the models were calculated using the area under the receiver operating curve (AUC) and calibration (i.e. the agreement between predicted and observed outcome) was assessed with the Hosmer-Lemeshow "goodness-of-fit" test. Overall model performance of the 3D-CRT based predictive models for XER(6M) and STIC(6M) was significantly worse in terms of the Brier score and R(2) Nagelkerke among patients treated with IMRT. Moreover the AUC for both 3D-CRT based models in the IMRT treated patients were markedly lower. The Hosmer-Lemeshow test showed a significant disagreement for both models between predicted risk and observed outcome. 3D-CRT based models for patient-rated xerostomia and sticky saliva among head and neck cancer patients treated with primary radiotherapy or chemoradiation turned out to be less valid for patients treated with IMRT. The main message from these findings is that models developed in a population treated with a specific technique cannot be generalised and extrapolated to a population treated with another technique without external validation.


Evaluation of epidural analgesia.
Lumbar epidural analgesia is outstandingly the best and most reliable method of pain relief in labour. It offers great advantages to both mother and child in cases of obstetric or pertinent non-obstetric abnormality. However, it demands care and attention, and requires a conduct of labour which in some cases is different from that to which most clinicians and midwives have been accustomed. It is necessary, therefore, that considerable experience be gained in the range of responses which may be evoked before the service is provided for mothers and infants who are at risk.


Resolution and recurrence rates of idiopathic trigger finger after corticosteroid injection.
This study addresses factors associated with apparent resolution and recurrence of triggering using data from providers with various treatment strategies. A retrospective review identified 878 adult patients with 1,210 Quinnell grade 2 or 3 trigger fingers that had one or more corticosteroid injections by one of three surgeons between 2001 and 2011. Two surgeons injected dexamethasone, but one had patients return 1 month after injection and was quick to recommend surgery (strategy A) and the other had patients return 2 months after injection, offered another injection or surgery, and followed the patient's preference. One surgeon used triamcinolone, had patients return only if the injection did not work, and waited at least 3 months to offer surgery. Factors associated with apparent resolution and recurrence of triggering were sought in bivariable and multivariable statistical analysis. Triamcinolone injection was associated with more frequent apparent resolution (83 %), than dexamethasone injection (30 %). Apparent resolution of triggering was also associated with a delayed surgery treatment strategy (B and C) and the affected finger (long and ring fingers were less likely to resolve). Return with triggering after documented or presumed resolution occurred in 188 fingers (33 %) and was associated with triamcinolone injection, index, long and ring finger, and orally treated non-insulin-dependent diabetes mellitus. Strategy A had the lowest initial apparent resolution rate, the highest proportion of patients having surgery, and the lowest final triggering rate of 10 %. Both treatment strategy and type of corticosteroid determine apparent resolution and recurrence rates.


Re-examining the relationship between audiometric profile and tinnitus pitch.
We explored the relationship between audiogram shape and tinnitus pitch to answer questions arising from neurophysiological models of tinnitus: 'Is the dominant tinnitus pitch associated with the edge of hearing loss?' and 'Is such a relationship more robust in people with narrow tinnitus bandwidth or steep sloping hearing loss?' A broken-stick fitting objectively quantified slope, degree and edge of hearing loss up to 16 kHz. Tinnitus pitch was characterized up to 12 kHz. We used correlation and multiple regression analyses for examining relationships with many potentially predictive audiometric variables. 67 people with chronic bilateral tinnitus (43 men and 24 women, aged from 22 to 81 years). In this ample of 67 subjects correlation failed to reveal any relationship between the tinnitus pitch and the edge frequency. The tinnitus pitch generally fell within the area of hearing loss. The pitch of the tinnitus in a subset of subjects with a narrow tinnitus bandwidth (n = 23) was associated with the audiometric edge. Our findings concerning subjects with narrow tinnitus bandwidth suggest that this can be used as an a priori inclusion criterion. A large group of such subjects should be tested to confirm these results.


Quantitative ultrastructural effects of cisplatin (Platinol), carboplatin (JM8), and iproplatin (JM9) on neurons of freshwater snail Lymnaea stagnalis.
Qualitative and quantitative ultrastructural effects of the platinum compounds cisplatin (Platinol), carboplatin (JM8), and iproplatin (JM9) were studied on two types of identified peptidergic neuron (caudodorsal cells, light green cells) in the pond snail Lymnaea stagnalis. Depending on the parameter under investigation, either one or both cell types were studied. Central nervous systems of the snail were incubated for 5 and 20 h in various identical and equitoxic drug concentrations. Cisplatin had the most severe effects. Platinol, i.e., cisplatin dissolved in NaCl solution with the addition of HCl (pH 2.0-3.0), as well as cisplatin dissolved in snail Ringer's solution (pH 7.8), caused swelling of axons and distensions of the intercellular spaces. This drug induced an increase in chromatin clump size in the caudodorsal cells (20-h incubation), while carboplatin and iproplatin induced the formation of many small chromatin clumps. Incubation in snail Ringer's solution (controls) and cisplatin affect the morphology of the nucleoli. At high dosages of cisplatin, the nucleoli of light green cells were transformed into homogeneous dense structures. The data indicate that platinum compounds react with nuclear and nucleolar DNA. All three drugs affected the activity and organization of the rough endoplasmic reticulum and the Golgi apparatus of the peptidergic neurons studied (qualitative observations). These effects, which point to a reduced neuropeptide synthesis, may be secondary, i.e., exerted via inhibition of RNA synthesis and ribosome formation (nucleoli). The fact that the number of neuropeptide granules in the cytoplasm of the cells remained constant (both cell types) may indicate that granule transport was also inhibited. Cisplatin and iproplatin induced an increase in the number of lysosomes in the light green cells. The number of lipid droplets in these cells was not affected by drug treatment. The results corroborate clinical data indicating that cisplatin is highly neurotoxic. Despite conflicting clinical data, observations on the snail neurons suggest that iproplatin is also neurotoxic, although less than cisplatin. Carboplatin is minimally neurotoxic, which is in accordance with clinical data. The central nervous system of Lymnaea is a suitable model for studying possible neurotoxic effects of platinum compounds.


Analysis of morphological and functional maturation of neoislets generated in vitro from pancreatic ductal cells and their suitability for islet banking and transplantation.
The pancreatic ductal stem cells are known to differentiate into islets of Langerhans; however, their yield is limited and the islet population is not defined. Therefore, the aims of the present study were to improvise a methodology for obtaining large numbers of islets in vitro and to characterize their morphological and functional status for islet cell banking and transplantation. Pancreatic ductal epithelial cell cultures were set in serum-free medium. Monolayers of epithelial cells in culture gave rise to islet-like clusters within 3-4 weeks. The identity of neoislets was confirmed by dithizone staining and analysis of the gene expression for endocrine markers by reverse transcriptase-polymerase chain reaction (RT-PCR). The islet population obtained was analysed by image analysis and insulin secretion in response to secretagogues. The cellular extracts from neoislets were immunoreactive to anti-insulin antibody and expressed insulin, glucagon, GLUT-2, PDX-1 and Reg-1 genes. The islets generated within 3-4 weeks exhibited a mixed population of large- and small-sized islets with clear cut dichotomy in the pattern of their insulin secretion in response to L-arginine and glucose. These neoislets maintained their structural and functional integrity on cryopreservation and transplantation indicating their suitability for islet cell banking. Thus, the present study describes an improved method for obtaining a constant supply of large numbers of islets from pancreatic ductal stem cell cultures. The newly generated islets undergo functional maturation indicating their suitability for transplantation.


Random selection of points distributed on curved surfaces.
Monte Carlo simulation is widely used in medical physics to obtain realistic solutions to radiation transport problems involving complex geometries. An important step is the random selection of the initial point in space from which the primary photon or charged particle originates. In many practical cases, the primary-particle sites are distributed over curved surfaces. We describe general sampling techniques that can be used to randomly choose the trajectory origin for sources distributed over any smooth surface that can be parametrically represented as a function of two variables. The source distribution need not be uniform. Implementation of these methods is particularly straight-forward for surfaces enclosing simple convex volumes such as spheres and ellipsoids.


Efficacy of Single-dose and 2-dose Intravenous Administration of Ramosetron in Preventing Postoperative Nausea and Vomiting After Laparoscopic Gynecologic Operation: A Randomized, Double-blind, Placebo-controlled, Phase 2 Trial.
This randomized trial investigated whether a 2-dose administration of intravenous ramosetron (5-hydroxytryptamine type 3 receptor antagonist) is more effective than a single-dose administration in preventing postoperative nausea and vomiting (PONV) in 89 patients who were scheduled to undergo laparoscopic operation for benign gynecologic diseases and to receive intravenous patient-controlled analgesia for relief of postoperative pain. After assignment at a ratio of 1:1, intravenous ramosetron (0.3 mg) was initially administered at the end of skin closure in all patients. Thereafter, ramosetron (0.3 mg) and placebo were administered to the study and control groups, respectively, at 4 hours after the operation. The baseline and operative characteristics were similar between the groups. The incidence of PONV during the 24-hour period after operation which was assessed as the primary endpoint did not differ between the groups. No serious adverse events occurred in either group. A 2-dose administration of intravenous ramosetron may not be superior to a single-dose administration in preventing PONV in patients undergoing laparoscopic operation for benign gynecologic diseases.


Detection of the KIT D816V mutation in peripheral blood of systemic mastocytosis: diagnostic implications.
Recent studies have found the KIT D816V mutation in peripheral blood of virtually all adult systemic mastocytosis patients once highly sensitive PCR techniques were used; thus, detection of the KIT D816V mutation in peripheral blood has been proposed to be included in the diagnostic work-up of systemic mastocytosis algorithms. However, the precise frequency of the mutation, the biological significance of peripheral blood-mutated cells and their potential association with involvement of bone marrow hematopoietic cells other than mast cells still remain to be investigated. Here, we determined the frequency of peripheral blood involvement by the KIT D816V mutation, as assessed by two highly sensitive PCR methods, and investigated its relationship with multilineage involvement of bone marrow hematopoiesis. Overall, our results confirmed the presence of the KIT D816V mutation in peripheral blood of most systemic mastocytosis cases (161/190; 85%)--with an increasing frequency from indolent systemic mastocytosis without skin lesions (29/44; 66%) to indolent systemic mastocytosis with skin involvement (124/135; 92%), and more aggressive disease subtypes (11/11; 100%)--as assessed by the allele-specific oligonucleotide-qPCR method, which was more sensitive (P<.0001) than the peptide nucleic acid-mediated PCR approach (84/190; 44%). Although the presence of the KIT mutation in peripheral blood, as assessed by the allele-specific oligonucleotide-qPCR technique, did not accurately predict for multilineage bone marrow involvement of hematopoiesis, the allele-specific oligonucleotide-qPCR allele burden and the peptide nucleic acid-mediated-PCR approach did. These results suggest that both methods provide clinically useful and complementary information through the identification and/or quantification of the KIT D816V mutation in peripheral blood of patients suspected of systemic mastocytosis.


Gender differences in the relationship between perceived discrimination and personality traits in young adulthood: Evidence using sibling fixed effects.
Although perceived discrimination (PD) is known to be associated with personality traits, family background characteristics may confound this association. Moreover, little is known about whether the relationship differs by gender. This study investigates whether the association between PD and personality traits is confounded by family background characteristics. Given gender differences in contexts and perceptions of discrimination as well as personality traits, this study also explores whether the association between PD and personality traits differs for men and women. Using data from the National Longitudinal Study of Adolescent to Adult Health, this study examines the association between PD and Big Five personality traits among young adults. This study uses sibling fixed effects models with a lagged dependent variable to account for unobservable family-level characteristics, such as genetics, parental characteristics, family environment, and childhood social contexts. Sibling fixed effects estimates showed that PD was associated with lower levels of conscientiousness and extraversion and higher levels of neuroticism. There were also gender differences such that PD was associated with lower conscientiousness only for women and lower extraversion only for men. The positive association with neuroticism was apparent for both men and women. This study suggests that the association between PD and personality traits is generally not confounded by stable family-level characteristics shared by siblings. This study also documents gender differences in the relationship between PD and personality traits. Given substantial implications of personality for a broad range of outcomes, especially among young adults, the findings of this study reaffirm the commitment of the whole society to eradicate any form of discrimination.


Hypothesis: Beta-adrenergic receptor blockers and weight gain: A systematic analysis.
One of the arguments put forward against the primary use of beta-blockers has been concern about adverse metabolic effects, such as unfavorable effects on lipids or insulin sensitivity. Another less-appreciated potential drawback is their propensity to cause weight gain in some patients. In 8 evaluable prospective randomized controlled trials that lasted >/=6 months, body weight was higher in the beta-blocker than in the control group at the end of the study. The median difference in body weight was 1.2 kg (range -0.4 to 3.5 kg). A regression analysis suggested that beta-blockers were associated with an initial weight gain during the first few months. Thereafter, no further weight gain compared with controls was apparent. There was no relationship between demographic characteristics and changes in body weight. Based on these observations, the first-line use of beta-blockers in obese hypertensive patients should be reviewed. Obesity management in overweight hypertensive patients may be more difficult in the face of beta-blocker treatment.


Current status of heroic limb salvage for critical limb ischemia.
The treatment of chronic lower extremity peripheral arterial disease is in a state of flux. During the past decade, vascular surgeons have assumed the responsibility for the endovascular care of patients needing vascular intervention. Once surgeons began performing these procedures, a change in attitude toward angioplasty resulted in an "endovascular explosion" and an overall reassessment of the traditional approaches to critical limb ischemia. Our current method of assessing outcomes is also in a state of flux. The original measure of procedural success, reconstruction patency, has been found to be a poor predictor of both patient palliation and functional success. A shift toward determining more accurate, patient-oriented outcome measures is ongoing. Until then, evidence would suggest that there are patients with such severe medical comorbidities, which include impaired ambulatory ability at presentation, that the benefits of revascularization seem to be insignificant. As our patient population ages and our healthcare system continues to fail financially, economic rationing motivated by lack of evidence-based data to the contrary may dictate that these patients are best served by primary limb amputation.


Association mapping for chilling tolerance in elite flint and dent maize inbred lines evaluated in growth chamber and field experiments.
Chilling sensitivity of maize is a strong limitation for its cultivation in the cooler areas of the northern and southern hemisphere because reduced growth in early stages impairs on later biomass accumulation. Efficient breeding for chilling tolerance is hampered by both the complex physiological response of maize to chilling temperatures and the difficulty to accurately measure chilling tolerance in the field under fluctuating climatic conditions. For this research, we used genome-wide association (GWA) mapping to identify genes underlying chilling tolerance under both controlled and field conditions in a broad germplasm collection of 375 maize inbred lines genotyped with 56 110 single nucleotide polymorphism (SNP). We identified 19 highly significant association signals explaining between 5.7 and 52.5% of the phenotypic variance observed for early growth and chlorophyll fluorescence parameters. The allelic effect of several SNPs identified for early growth was associated with temperature and incident radiation. Candidate genes involved in ethylene signalling, brassinolide, and lignin biosynthesis were found in their vicinity. The frequent involvement of candidate genes into signalling or gene expression regulation underlines the complex response of photosynthetic performance and early growth to climatic conditions, and supports pleiotropism as a major cause of co-locations of quantitative trait loci for these highly polygenic traits.


Sociality in the gray mouse lemur (Microcebus murinus) in northwestern Madagascar.
The mating system and social organization of the gray mouse lemur (Microcebus murinus) was investigated in two three-month field studies (covering a period before, during, and after the first mating season) in Ampijoroa, northwestern Madagascar. The spatial and temporal distribution of the sexes within a population was studied using mark/recapture techniques and radiotelemetry to assess possible contest or scramble competition between the males. Sociality was inferred from the occurrence and probability of nocturnal social encounters, the temporal stability of daily sleeping groups, and nocturnal ranging patterns of co-sleepers. Males and females were evenly distributed in the study area within a network of highly overlapping home ranges. No indications were found for the spatial monopolization of the females by certain dominant males. Males and females had spatial access to several potential mates; the mating system is therefore characterized as a multi-male/multi-female system. Male home range sizes increased during the first mating season, which was interpreted as an indicator for scramble competition between the males. Competitive mate searching, sperm competition, and temporary mate guarding as well as female mate choice are suggested as the most probable reproductive strategies. Over the course of the study the animals lived continually within the study area, and most females formed stable individualized sleeping associations. Females that slept together shared a higher percentage of their home range than did females that slept at different sites. It is suggested that this network of social relationships should be described as a dispersed but individualized neighborhood.