Ubiquitin and phosphorylated neurofilament epitopes in ballooned neurons of the extraocular muscle nuclei in a case of Werdnig-Hoffmann disease.
The extraocular muscle nuclei in one case of Werdnig-Hoffmann disease were examined immunocytochemically using antibodies against phosphorylated neurofilament (pNF) and ubiquitin (UBQ). The oculomotor and trochlear nuclei showed several chromatolytic ballooned neurons. All ballooned neurons contained epitopes of pNF and UBQ. pNF were present mainly in the periphery of the cell in a ring-like shape and were occasionally seen in the center of some cells. On the other hand, the structures stained by the antibody to UBQ were small vesicles or granules and most of them were aggregated in the center of the cell. These distribution patterns of pNF and UBQ may be unique in Werdnig-Hoffmann disease, since similar patterns were reported in other types of neurons of Werdnig-Hoffmann disease but were not seen in two other motor neuron diseases: classical amyotrophic lateral sclerosis, and familial amyotrophic lateral sclerosis with posterior column and spinocerebellar tract involvement.


Acceptability and psychological impact of out-of-office monitoring to diagnose hypertension: an evaluation of survey data from primary care patients.
Out-of-office blood pressure (BP) is recommended for diagnosing hypertension in primary care due to its increased accuracy compared to office BP. Moreover, being diagnosed as hypertensive has previously been linked to lower wellbeing. There is limited evidence regarding the acceptability of out-of-office BP and its impact on wellbeing. To assess the acceptability and psychological impact of out-of-office monitoring in people with suspected hypertension. A pre- and post-evaluation of participants with elevated (≥130 mmHg) systolic BP, assessing the psychological impact of 28 days of self-monitoring followed by ambulatory BP monitoring for 24 hours. Participants completed standardised psychological measures pre- and post-monitoring, and a validated acceptability scale post-monitoring. Descriptive data were compared using χ2 tests and binary logistic regression. Pre- and post-monitoring comparisons were made using the paired t-test and Wilcoxon signed rank test. Out-of-office BP monitoring had no impact on depression and anxiety status in 93% and 85% of participants, respectively. Self-monitoring was more acceptable than ambulatory monitoring (n = 183, median 2.4, interquartile range [IQR] 1.9-3.1 versus median 3.2, IQR 2.7-3.7, P<0.01). When asked directly, 48/183 participants (26%, 95% confidence interval [CI] = 20 to 33%) reported that self-monitoring made them anxious, and 55/183 (30%, 95% CI = 24 to 37%) reported that ambulatory monitoring made them anxious. Out-of-office monitoring for hypertension diagnosis does not appear to be harmful. However, health professionals should be aware that in some patients it induces feelings of anxiety, and self-monitoring may be preferable to ambulatory monitoring.


Entanglement spectrum of a disordered topological Chern insulator.
We investigate the behavior of a topological Chern insulator in the presence of disorder, with a focus on its entanglement spectrum (EtS) constructed from the ground state. For systems with symmetries, the EtS was shown to contain explicit information about the topological universality class revealed by sorting the EtS against the conserved quantum numbers. In the absence of any symmetry, we demonstrate that statistical methods such as the level statistics of the EtS can be equally insightful, allowing us to distinguish when an insulator is in a topological or trivial phase and to map the boundary between the two phases. The phase diagram of a Chern insulator is explicitly computed as function of Fermi level (EF) and disorder strength using the level statistics of the EtS and energy spectrum, together with a computation of the Chern number (C) via a new, efficient real-space formula.


Interferon beta-1b in the treatment of multiple sclerosis: final outcome of the randomized controlled trial. The IFNB Multiple Sclerosis Study Group and The University of British Columbia MS/MRI Analysis Group.
Our previously reported multicenter, blinded, randomized, controlled study of two doses of interferon beta-1b (IFNB) in 372 patients demonstrated a reduction in relapse frequency and severity and in MRI activity. We now report the results of the continuation of that study. The median time on study was 46.0 months for the placebo arm, 45.0 months for 1.6 million international units (MIU), and 48.0 months for 8 MIU. IFNB had a persistent beneficial effect on exacerbation rate and MRI burden of disease and was relatively free of long-term side effects. There was a one-third reduction in exacerbation rate in the 8-MIU treatment arm, compared with placebo, in each of 5 years. Serial annual MRI was done in all patients, and 217 of the patients had either a fourth- or fifth-year scan. There was no significant progression of lesion burden in the 8-MIU arm in each successive year compared with baseline (at 4 years, p = 0.917), whereas a highly significant increase in lesion area occurred in the placebo arm (p = 0.0001). Among the 154 noncompleters, there was no systematic bias recognized that favored either treatment arm for the outcome measures of exacerbation rate, disability, or MRI activity. Dropouts in the placebo group had higher exacerbation rates and accumulation of MRI lesion burden than did dropouts in the other treatment arms, which probably reduced the power of the study to demonstrate treatment effects on these measures in the later years of the trial.(ABSTRACT TRUNCATED AT 250 WORDS)


Unemployment and substance use: a review of the literature (1990-2010).
The current article summarizes the results of a comprehensive review of the international research published between 1990 and 2010. The research was focused on the prevalence of substance use/disorders among the unemployed and employed, the impact of substance abuse on unemployment and vice versa, the effect of unemployment on alcohol/ drug addiction treatment and smoking cessation, and the relationship between business cycle, unemployment rate and substance use. Over hundred-thirty relevant studies were identified investigating these issues. The main results are as follows: (1) Risky alcohol consumption (associated with hazardous, binge, and heavy drinking) is more prevalent among the unemployed. They are also more likely to be smokers, to use illicit and prescription drugs, and to have alcohol and drug disorders (abuse, dependence). (2) Problematic substance use increases the likelihood of unemployment and decreases the chance of finding and holding down a job. (3) Unemployment is a significant risk factor for substance use and the subsequent development of substance use disorders. However, the current research provides only limited information about which individuals are more likely to be affected. (4) Unemployment increases the risk of relapse after alcohol and drug addiction treatment. (5) The exact nature of the relationship between unemployment and the probability of smoking cessation remains unclear due to the mixed results observed in the literature review. (6) Drinking and smoking patterns appear to be procyclical. We see a decrease in both when the economy declines and the unemployment rate increases. In contrast, a countercyclical trend was observed amongst adolescent drug users. However, these studies do not provide any convincing or additional information about substance use amongst the unemployed. This paper discusses the merits, limitations and problems of the research, proposes numerous future research questions, and outlines important implications for policy makers and practitioners, especially with regard to prevention and vocational promotion and rehabilitation.


Nurses' views of the coping and support mechanisms experienced in managing terminal haemorrhage.
Although palliative care is a rewarding specialty, it presents emotive personal challenges for the health professionals working within it. Terminal haemorrhage is arguably the most feared and distressing event in the palliative care setting, both for the patient and for the health professionals looking after them. The aim of this study was to explore and reflect on the coping and support mechanisms that have helped nurses to manage these events. Purposive sampling was used to recruit nurses from palliative care and oncology who had personal experience of managing terminal haemorrhage. Semi-structured interviews were used to collect qualitative data that were then analysed using interpretative phenomenological analysis. Saturation of data was reached after 11 participants had been interviewed. The participants had a combined experience of managing 37 patients with a terminal haemorrhage. The key themes that emerged were the role of the 'autopilot', having a plan, education and training, the value of debriefing and peer support, and the importance of supporting the whole team. Education and training specifically about terminal haemorrhage and structured debriefing after such events were both identified by the participants as currently unmet needs. Professionals working in areas in which patients are at particular risk of terminal haemorrhage require adequate training and education prior to these events and a structure for formal debriefing and peer support afterwards.


DNA polymerase requirements for parvovirus H-1 DNA replication in vitro.
An in vitro system using nuclei from parvovirus H-1-infected cells was used to characterize the influence of inhibitors of mammalian DNA polymerases on viral DNA synthesis. The experiments tested the effects of aphidicolin, which is highly specific for DNA polymerase alpha, and 2',3'-dideoxythymidine-5'-triphosphate (ddTTP), which inhibits cellular DNA polymerases in the order gamma greater than beta greater than alpha. Both aphidicolin and ddTTP were inhibitory, indicating that both polymerase alpha and a ddttp-sensitive enzyme are required for viral DNA synthesis. This was seen more clearly in kinetic measurements, which indicated an initial period of rapid DNA synthesis with the participation of polymerase alpha, followed by a period of less rapid, but more sustained, rate of DNA synthesis carried out by a ddTTP-sensitive enzyme, probably polymerase gamma. One interpretation of the results is that polymerase alpha functions in a strand displacement stage of the viral DNA replication mechanism, whereas polymerase gamma serves to convert the displaced single strands back to double-strand replicative form.


Cross-sectional comparisons of BMI and waist circumference in British children: mixed public health messages.
Research suggests that there has been a leveling off in obesity prevalence occurring in the child population. However, a concern with the evidence base is that all of the studies have relied upon the use of BMI. The purpose of this study was to compare waist circumference (WC), BMI, and waist-to-height ratio (WHtR) data in three different sample of children (total number: 14,697) typically aged 11-12 years. Obesity prevalence defined by BMI did not change significantly between measurement years (2005 boys 20.6%, girls 18.0%; 2006 boys 19.3%, girls 17.3%; 2007 boys 19.8%, girls 16.4%). Obesity prevalence defined by WC was considerably higher especially, in girls (2005 boys 26.3%, girls 35.6%; 2006 boys 20.3%, girls 28.2%; 2007 boys 22.1%, girls 30.1%). The prevalence of children defined as "at risk" according to WHtR (2005 boys 23.3%, girls 21.1%; 2006 boys 16.7%, girls 15.6%; 2007 boys 17.6%, girls 17.2%) was found to be between obesity prevalence, estimated using BMI and WC. This data are the most up to date collection that includes BMI and WC in three large samples of children and clearly demonstrates inconsistencies between different measurements based on current classification systems. There is a need to understand the relationship between BMI and WC, with growth and health risk to establish a consistent public health message that is easily understood by the public.


Role of Fine Needle Aspiration Cytology (FNAC) as a Diagnostic Tool in Paediatric Head and Neck Lymphodenopathy.
A neck mass that is present for longer than a week might be pathological requiring rapid and thorough evaluation. This study aims to evaluate the positive role Fine needle aspiration cytology plays in the diagnosis of pediatric patients with lymphadenopathy in the head and neck region. Fine needle aspiration of lymph nodes was carried out, fixed and stained by cytopathologists for 56 patients at the Jos University Teaching Hospital, 43 (76.8%) were inflammatory and 13 (23.2%) malignant. Reactive hyperplasia (72.1%), acute suppurative (18.6%) and tuberculosis (9.3%) constituted the inflammatory lesions. Non Hodgkins lymphoma was the commonest malignant lesion (76.9%) followed by Hodgkins lymphoma (15.4%), the least being Rhabdomyosarcoma (7.7%). Age range 10-14 years had the highest number of cases (46.4%). Males were 36 (64.3%) while females were 20 (35.7%). All malignant cases diagnosed by FNAC had to undergo confirmation/characterization by histology and had 100% concordance. Thus there were no false positives and specificity was 100%, sensitivity 100%. Of the 43 diagnosed as inflammatory by FNAC, 12 cases which did not resolve after treatment or where patients condition worsened had to undergo surgical biopsy. Out of these only 1 (8.3%) case of fungal infection was misdiagnosed by FNAC. The lymph nodes were generalized 4 (7.1%) and localized in 52 (92.9%). Maximum number of cases 53 (94.6%) had Cervical Lymphadenopathy followed by axillary 2 (3.6%) and inguinal 1 (1.8%). Out of the cervical group of nodes, the upper anterior and upper posterior deep cervical nodes were involved in majority of cases (95%).


Preceptors' strategies for correcting residents in an ambulatory care medicine setting: a qualitative analysis.
To understand the interactional strategies preceptors use as they relate to and occasionally correct interns in a general internal medicine teaching clinic. An observational, cohort study was carried out from May 1990 through May 1993 of the precepting conversations between 11 pairs of interns and faculty preceptors in the general internal medicine ambulatory care clinic of the Hospital of the University of Pennsylvania. All interactions were videotaped and transcribed. The interactions provided numerous examples of preceptors' correcting of interns. These were analyzed qualitatively by ethnographic and conversation-analytic methods. The strategies the preceptors used to correct the interns were complicated and quite indirect, and tended to minimize exposing the interns' errors. These strategies revealed the dilemmas inherent in being a preceptor and also the beliefs the preceptors brought to their task. The preceptors' strategies demonstrated their high regard for maintaining the interns' self-esteem and sense of responsibility, as well as the preceptors' willingness to forego, at least for the moment, correctional strategies that might have been more explicit and direct. The preceptors' general approach to correcting interns was consistent with pedagogic norms favoring discovery learning and with societal norms favoring egalitarianism and respect for individuals. This approach, however, may not be free of problems, and raises questions regarding the effects such strategies have upon the interns' capacity for accurate self-assessment, including the assessment of their own knowledge bases.


Change in surface roughness by dynamic shape-memory acrylate networks enhances osteoblast differentiation.
Microscale surface roughness has been shown to enhance osseointegration of titanium implants through increased osteoblast differentiation while osteoblast proliferation remains greater on smooth titanium. Taking advantage of these phenomena, we developed a shape memory (meth)acrylate copolymer with thermomechanical properties that created a time-dependent dynamic surface change from smooth to rough under in vitro cell culture conditions and evaluated the effect of the shape recovery on osteoblast response. Rough topographies were created using soft lithography techniques to mimic those found on clinically-used Ti surfaces (machined smooth; acid-etched; grit-blasted). The surface roughness was then reduced to smooth via compression and shown to fully recover within 24 h in culture conditions. When grown under static conditions, osteoblast number, alkaline phosphatase specific activity (ALP), and osteoprotegerin (OPG) and vascular endothelial growth factor (VEGF) production were unaffected by polymer surface roughness, but osteocalcin (OCN) was increased on the grit-blasted polymer mimic. Under dynamic conditions, DNA was reduced but OCN and OPG were increased on the compressed grit-blasted polymer at 3 days compared to static surfaces. The present study indicates that responses to polymer surface are sensitive to time-dependent changes in topography. The use of a shape memory polymer with dynamic surface roughness may improve osseointegration.


PKC1 is essential for protection against both oxidative and nitrosative stresses, cell integrity, and normal manifestation of virulence factors in the pathogenic fungus Cryptococcus neoformans.
Cell wall integrity is crucial for fungal growth, survival, and pathogenesis. Responses to environmental stresses are mediated by the highly conserved Pkc1 protein and its downstream components. In this study, we demonstrate that both oxidative and nitrosative stresses activate the PKC1 cell integrity pathway in wild-type cells, as measured by phosphorylation of Mpk1, the terminal protein in the PKC1 phosphorylation cascade. Furthermore, deletion of PKC1 shows that this gene is essential for defense against both oxidative and nitrosative stresses; however, other genes involved directly in the PKC1 pathway are dispensable for protection against these stresses. This suggests that Pkc1 may have multiple and alternative functions other than activating the mitogen-activated protein kinase cascade from a "top-down" approach. Deletion of PKC1 also causes osmotic instability, temperature sensitivity, severe sensitivity to cell wall-inhibiting agents, and alterations in capsule and melanin. Furthermore, the vital cell wall components chitin and its deacetylated form chitosan appear to be mislocalized in a pkc1Delta strain, although this mutant contains wild-type levels of both of these polymers. These data indicate that loss of Pkc1 has pleiotropic effects because it is central to many functions either dependent on or independent of PKC1 pathway activation. Notably, this is the first time that Pkc1 has been implicated in protection against nitrosative stress in any organism.


An Evidence-Based Systematic Review on the Diagnostic Accuracy of Hearing Screening Instruments for Preschool- and School-Age Children.
The purpose of this article is to conduct an evidence-based systematic review on the accuracy of pure-tone or otoacoustic emission (OAE) screening for identifying hearing loss in preschool- and school-age children. A systematic search of the literature published between 1975 and 2013 was conducted. Articles meeting the selection criteria were critically appraised for quality. Selection criteria required that behavioral thresholds be measured in children failing the screen and in at least a subset of children passing the screen. Sensitivity and specificity were used to calculate positive and negative likelihood ratios that could be compared between instruments. Eighteen studies were included in the final analysis. There was considerable variability among studies on stimulus levels, response criteria, and definition of hearing loss. Approximately half of positive and negative likelihood ratio pairs for OAEs (52%) and pure-tone screening (45%) were considered suggestive or informative for identifying hearing loss. Both pure-tone and OAE screening can identify hearing loss in preschool- and school-age children. Studies that compared both tools in the same population concluded that pure-tone screening had higher sensitivity than OAE screening and thus was considered the preferred tool. Future research should incorporate standard stimulus levels, response criteria, and definitions of hearing loss.


The evolution of proton beam therapy: Current and future status.
Proton beam therapy (PBT) has been increasingly used in a variety of cancers due to its excellent physical properties and superior dosimetric parameters. PBT may improve patient survival by improving the local tumor treatment rate while reducing injury to normal organs, which may result in fewer radiation-induced adverse effects. However, the significant cost of establishing and maintaining proton facilities cannot be overlooked. In addition, there has been significant controversy regarding routine application of this treatment in certain types of cancer. The challenges of PBT in the future mainly include the lack of basic clinical trials, unclear biological effects, immature imaging technology and miniaturization of imaging guidance. Overcoming these limitations may promote the rapid development of PBT. We herein provide an overview of the existing literature on the efficacy and toxicity of common oncological applications of proton beam therapy.


Intracellular trafficking and assembly of specific Kir3 channel/G protein complexes.
We have previously demonstrated that Kir3.1 channels and Gbeta1gamma2 subunits initially interact in the endoplasmic reticulum (ER). To elucidate the role that anterograde protein trafficking pathways may play in the formation of these complexes, we used dominant negative (DN) mutants of the small G proteins Sar 1 and various compartment-specific Rabs which impede anterograde protein trafficking at different steps. Sar 1 H79G and Rab 1 S25N mutants efficiently blocked the plasma membrane trafficking of the Kir3.1/Kir3.4 complex however they did not block the Gbeta1gamma2/Kir3.1 interaction as measured using bioluminescence resonance energy transfer (BRET). This interaction was also insensitive to the presence of DN Rabs 2, 6, 8, and 11. These results confirm that Gbetagamma/Kir3 complexes form early during channel biosynthesis and trafficking. Using a combination of BRET, protein complementation assays and co-immunoprecipitation, we demonstrate that Gbeta1-4 can interact with Kir3.1 in the absence of Kir3.4. Gbeta5 does not directly interact with the channel but can still be co-immunoprecipitated as part of a larger complex. The interaction between Gbeta and Kir3.1 was selectively fostered by co-expression with different Ggamma subunits. When Ggamma1 or Ggamma11 was co-expressed with eGFP-Gbeta3 or eGFP-Gbeta4, the interaction with the effector was lost. Kir3.2 was capable of interacting with Gbeta1-3 and not Gbeta4 or Gbeta5. These interactions were again fostered by co-expression with Ggamma and were also insensitive to DN Sar 1 or Rab 1. Taken together, our data show that these "precocious" channel/G protein interactions are specific and may have implications beyond their basic function in signalling events.


High coherence plane breaking packaging for superconducting qubits.
We demonstrate a pogo pin package for a superconducting quantum processor specifically designed with a nontrivial layout topology (e.g., a center qubit that cannot be accessed from the sides of the chip). Two experiments on two nominally identical superconducting quantum processors in pogo packages, which use commercially available parts and require modest machining tolerances, are performed at low temperature (10 mK) in a dilution refrigerator and both found to behave comparably to processors in standard planar packages with wirebonds where control and readout signals come in from the edges. Single- and two-qubit gate errors are also characterized via randomized benchmarking, exhibiting similar error rates as in standard packages, opening the possibility of integrating pogo pin packaging with extensible qubit architectures.


Detection of intestinal metaplasia in Barrett's esophagus: an observational comparator study suggests the need for a minimum of eight biopsies.
Intestinal metaplasia (IM) and dysplasia in Barrett's esophagus are recognized surrogates for esophageal adenocarcinoma risk. While few would argue with the "hunt for dysplasia," there is a divide regarding the usefulness of the histological confirmation of intestinal metaplasia in endoscopically apparent long segment Barrett's esophagus. We aimed to assess the frequency of intestinal metaplasia in 125 consecutive patients with columnar-lined esophagus and to determine the optimal biopsy protocol to detect intestinal metaplasia. Two-hundred ninety-six endoscopies were performed over a 4-yr period in Barrett's esophagus segments of mean length 4 cm (range 1-11 cm) at a single center and the resulting biopsies were analyzed retrospectively. Biopsies were all processed with routine hematoxylin and eosin (H&E) staining, and a subset (N=92) was subject to alcian blue/periodic-acid Schiff staining. Using H&E staining, we found that the optimum number of biopsies to diagnose intestinal metaplasia was 8 per endoscopy, mean 67.9% endoscopies having intestinal metaplasia. In contrast, if only four were taken the yield was 34.7% with intestinal metaplasia. Unless more than 16 biopsies were taken (100% yield of intestinal metaplasia), no additional significant detection was achieved. Using additional alcian blue/periodic-acid Schiff staining only had a marginal benefit, with 5.4% of new cases of intestinal metaplasia being identified. There is a proximal cephalo-caudal gradient of intestinal metaplasia, especially with increased chronological age, but doing repeat endoscopies on patients did not increase the detection of intestinal metaplasia. The data suggest that at least 8 random biopsies is the minimum to be taken and analyzed with conventional H&E staining to diagnose benign intestinal metaplasia. Taking more biopsies did not statistically increase the diagnosis of intestinal metaplasia except when greater than 16 were taken when 100% yield was obtained.


Coarse-grained kinetic computations for rare events: application to micelle formation.
We discuss a coarse-grained approach to the computation of rare events in the context of grand canonical Monte Carlo (GCMC) simulations of self-assembly of surfactant molecules into micelles. The basic assumption is that the computational system dynamics can be decomposed into two parts-fast (noise) and slow (reaction coordinates) dynamics, so that the system can be described by an effective, coarse-grained Fokker-Planck (FP) equation. While such an assumption may be valid in many circumstances, an explicit form of FP equation is not always available. In our computations we bypass the analytic derivation of such an effective FP equation. The effective free energy gradient and the state-dependent magnitude of the random noise, which are necessary to formulate the effective Fokker-Planck equation, are obtained from ensembles of short bursts of microscopic simulations with judiciously chosen initial conditions. The reaction coordinate in our micelle formation problem is taken to be the size of a cluster of surfactant molecules. We test the validity of the effective FP description in this system and reconstruct a coarse-grained free energy surface in good agreement with full-scale GCMC simulations. We also show that, for very small clusters, the cluster size ceases to be a good reaction coordinate for a one-dimensional effective description. We discuss possible ways to improve the current model and to take higher-dimensional coarse-grained dynamics into account.


Hippocampal and cortical growth-associated protein-43 messenger RNA in schizophrenia.
Growth-associated protein-43 is involved in maturational and plasticity-associated processes, and changes in growth-associated protein-43 expression are a marker of altered plasticity following experimental and neuropathological lesions. Using in situ hybridization, we have investigated growth-associated protein-43 mRNA in the medial temporal lobe and cerebral cortex in 11 normal subjects and 11 matched subjects with schizophrenia, a disorder in which perturbed neurodevelopment and aberrant plasticity are implicated. In the schizophrenia group, growth-associated protein-43 messenger RNA was decreased in the medial temporal lobe, primary visual cortex and anterior cingulate gyrus, but was unaltered in the superior temporal and dorsolateral prefrontal cortices. Correlations of growth-associated protein-43 messenger RNA signal between areas were stronger and more numerous in the schizophrenics than in the controls, suggesting a more global regulation of growth-associated protein-43 expression. Finally, the ratio of growth-associated protein-43 messenger RNA to synaptophysin messenger RNA--a putative index of the production of new synapses--was decreased in the medial temporal lobe in the schizophrenics. Our findings imply that neuronal plasticity as indexed by growth-associated protein-43 expression is impaired, and perhaps aberrantly regulated, in schizophrenia. The data support the emerging view that the disease pathophysiology is one which affects the hippocampal and cortical circuitry and that the abnormalities are reflected in the altered expression of specific neuronal genes.


Incidental non-Hodgkin's lymphoma in patients with localized prostate cancer.
To determine the outcome of patients with clinically organ-confined prostate cancer undergoing radical retropubic prostatectomy for cure and incidentally discovered concurrent low-grade non-Hodgkin's lymphoma at time of surgery. From September 1986 to September 1997, 4319 patients underwent radical retropubic prostatectomy at our institution. The records of 10 patients incidentally diagnosed to have low-grade non-Hodgkin's lymphoma at the time of radical prostatectomy were retrospectively reviewed. Of 4319 patients requiring radical prostatectomy, 10 (0.2%) were found to have low-grade non-Hodgkin's lymphoma. All 10 men had an uneventful postoperative course. Two patients subsequently developed progression of lymphoma, one of whom required treatment. One patient died of sepsis associated with his lymphoma and 1 patient died of an unrelated malignancy (lung cancer), both 7 years following surgery. Two patients developed biochemical prostate-specific antigen recurrence. The remainder of men were free of prostate cancer recurrence and experienced no progression of lymphoma at an average of 45 months (range 12 to 142). Patients with organ-confined prostate cancer, who are candidates for radical prostatectomy, experience long-term prostate cancer-free survival in the face of incidentally diagnosed low-grade lymphoma. Because the management of most incidentally discovered low-grade lymphomas is expectant, patients discovered at surgery to have this clinical entity should not be denied radical prostatectomy.


Mechanisms regulating melanogenesis.
Skin pigmentation is an important human phenotypic trait whose regulation, in spite of recent advances, has not yet been fully understood. The pigment melanin is produced in melanosomes by melanocytes in a complex process called melanogenesis. The melanocyte interacts with endocrine, immune, inflammatory and central nervous systems, and its activity is also regulated by extrinsic factors such as ultraviolet radiation and drugs. We have carried out a review of the current understanding of intrinsic and extrinsic factors regulating skin pigmentation, the melanogenesis stages and related gene defects. We focused on melanocyte-keratinocyte interaction, activation of melanocortin type 1 receptor (MC1-R) by peptides (melanocyte-stimulating hormone and adrenocorticotropic hormone) resulting from proopiomelanocortin (POMC) cleavage, and mechanisms of ultraviolet-induced skin pigmentation. The identification and comprehension of the melanogenesis mechanism facilitate the understanding of the pathogenesis of pigmentation disorders and the development of potential therapeutic options.


Molecular mechanisms of cannabinoid protection from neuronal excitotoxicity.
Cannabinoids protect neurons from excitotoxic injury. We investigated the mechanisms involved by studying N-methyl-D-aspartate (NMDA) toxicity in cultured murine cerebrocortical neurons in vitro and mouse cerebral cortex in vivo. The cannabinoid agonist R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)-methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl)-methanone mesylate [R(+)-Win 55212] reduced neuronal death in murine cortical cultures treated with 20 microM NMDA, and its protective effect was attenuated by the CB1 cannabinoid receptor (CB1R) antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-cichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride (SR141716A). Cultures from CB1R-knockout mice were more sensitive to NMDA toxicity than were cultures from wild-type mice. The in vitro protective effect of R(+)-Win 55212 was reduced by pertussis toxin, consistent with signaling through CB1R-coupled G-proteins. The nitric-oxide synthase (NOS) inhibitors 7-nitroindazole (7-NI) and N-omega-nitro-L-arginine methyl ester also reduced NMDA toxicity. In addition, CB1R and neuronal NOS were coexpressed in cultured cortical neurons, suggesting that cannabinoids might reduce NMDA toxicity by interfering with the generation of NO. NOS activity in cerebral cortex was higher in CB1R-knockouts than in wildtype mice, and 7-NI reduced NMDA lesion size. R(+)-Win 55212 inhibited NO production after NMDA treatment of wild-type cortical neuron cultures, measured with 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate, and this effect was reversed by SR141716A. In contrast, R(+)-Win 55212 failed to inhibit NO production in cultures from CB1R knockouts. Dibutyryl-cAMP blocked the protective effect of R(+)-Win 55212, and this was reversed by the protein kinase A (PKA) inhibitor N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide (H89). Cannabinoids seem to protect neurons against NMDA toxicity at least in part by activation of CB1R and downstream inhibition of PKA signaling and NO generation.


Evidence for a protein mutator in yeast: role of the Hsp70-related chaperone ssb in formation, stability, and toxicity of the [PSI] prion.
Propagation of the yeast protein-based non-Mendelian element [PSI], a prion-like form of the release factor Sup35, was shown to be regulated by the interplay between chaperone proteins Hsp104 and Hsp70. While overproduction of Hsp104 protein cures cells of [PSI], overproduction of the Ssa1 protein of the Hsp70 family protects [PSI] from the curing effect of Hsp104. Here we demonstrate that another protein of the Hsp70 family, Ssb, previously implicated in nascent polypeptide folding and protein turnover, exhibits effects on [PSI] which are opposite those of Ssa. Ssb overproduction increases, while Ssb depletion decreases, [PSI] curing by the overproduced Hsp104. Both spontaneous [PSI] formation and [PSI] induction by overproduction of the homologous or heterologous Sup35 protein are increased significantly in the strain lacking Ssb. This is the first example when inactivation of an unrelated cellular protein facilitates prion formation. Ssb is therefore playing a role in protein-based inheritance, which is analogous to the role played by the products of mutator genes in nucleic acid-based inheritance. Ssb depletion also decreases toxicity of the overproduced Sup35 and causes extreme sensitivity to the [PSI]-curing chemical agent guanidine hydrochloride. Our data demonstrate that various members of the yeast Hsp70 family have diverged from each other in regard to their roles in prion propagation and suggest that Ssb could serve as a proofreading component of the enzymatic system, which prevents formation of prion aggregates.


Screening for Adolescent Alcohol Use in the Emergency Department: What Does It Tell Us About Cannabis, Tobacco, and Other Drug Use?
The pediatric emergency department (PED) represents an opportune time for alcohol and drug screening. The National Institute of Alcohol Abuse and Alcoholism (NIAAA) recommends a two-question alcohol screen for adolescents as a predictor of alcohol and drug misuse. A multi-site PED study was conducted to determine the association between the NIAAA two-question alcohol screen and adolescent cannabis use disorders (CUD), cigarette smoking, and lifetime use of other drugs. Participants included 12-17-year olds (n = 4834) treated in one of 16 participating PEDs. An assessment battery, including the NIAAA two-question screen and other measures of alcohol, tobacco and drug use, was self-administered on a tablet computer. A diagnosis of CUD, lifetime tobacco use or lifetime drug use was predicted by any self-reported alcohol use in the past year, which indicates a classification of moderate risk for middle school ages and low risk for high school ages on the NIAAA two-question screen. Drinking was most strongly predictive of a CUD, somewhat weaker for lifetime tobacco use, and weakest for lifetime drug use. This same pattern held for high school and middle school students and was stronger for high school students over middle school students for all three categories. This association was also found across gender, ethnicity and race. The association was strongest for CUD for high school students, sensitivity 81.7% (95% CI, 77.0, 86.5) and specificity 70.4% (95% CI, 68.6, 72.1). Conclusions/Importance: A single question about past year alcohol use can provide valuable information about other substance use, particularly marijuana.


Biological pathway analysis by ArrayUnlock and Ingenuity Pathway Analysis.
Once a list of differentially expressed genes has been identified from a microarray experiment, a subsequent post-analysis task is required in order to find the main biological processes associated to the experimental system. This paper describes two pathways analysis tools, ArrayUnlock and Ingenuity Pathways Analysis (IPA) to deal with the post-analyses of microarray data, in the context of the EADGENE and SABRE post-analysis workshop. Dataset employed in this study proceeded from an experimental chicken infection performed to study the host reactions after a homologous or heterologous secondary challenge with two species of Eimeria. Analysis of the same microarray data source employing both commercial pathway analysis tools in parallel let to identify several biological and/or molecular functions altered in the chicken Eimeria maxima infection model, including several immune system related pathways. Biological functions differentially altered in the homologous and heterologous second infection were identified. Similarly, the effect of the timing in a homologous second infection was characterized by several biological functions. Functional analysis with ArrayUnlock and IPA provided information related to functional differences with the three comparisons of the chicken infection leading to similar conclusions. ArrayUnlock let an improvement of the annotations of the chicken genome adding InterPro annotations to the data set file. IPA provides two powerful tools to understand the pathway analysis results: the networks and canonical pathways that showed several pathways related to an adaptative immune response.


Transgene x environment interactions in genetically modified wheat.
The introduction of transgenes into plants may cause unintended phenotypic effects which could have an impact on the plant itself and the environment. Little is published in the scientific literature about the interrelation of environmental factors and possible unintended effects in genetically modified (GM) plants. We studied transgenic bread wheat Triticum aestivum lines expressing the wheat Pm3b gene against the fungus powdery mildew Blumeria graminis f.sp. tritici. Four independent offspring pairs, each consisting of a GM line and its corresponding non-GM control line, were grown under different soil nutrient conditions and with and without fungicide treatment in the glasshouse. Furthermore, we performed a field experiment with a similar design to validate our glasshouse results. The transgene increased the resistance to powdery mildew in all environments. However, GM plants reacted sensitive to fungicide spraying in the glasshouse. Without fungicide treatment, in the glasshouse GM lines had increased vegetative biomass and seed number and a twofold yield compared with control lines. In the field these results were reversed. Fertilization generally increased GM/control differences in the glasshouse but not in the field. Two of four GM lines showed up to 56% yield reduction and a 40-fold increase of infection with ergot disease Claviceps purpurea compared with their control lines in the field experiment; one GM line was very similar to its control. Our results demonstrate that, depending on the insertion event, a particular transgene can have large effects on the entire phenotype of a plant and that these effects can sometimes be reversed when plants are moved from the glasshouse to the field. However, it remains unclear which mechanisms underlie these effects and how they may affect concepts in molecular plant breeding and plant evolutionary ecology.


Mepacrine inhibits matrix metalloproteinases-1 (MMP-1) and MMP-9 activation in human fibroblast-like synoviocytes.
Matrix metalloproteinases (MMP) are enzymes known to be involved in normal physiological and in many pathological conditions. Rheumatic diseases are among the ailments where MMP have been shown to exert detrimental effects. Mepacrine is used alone or in combination with other drugs to treat lupus and other rheumatic diseases. We investigated whether mepacrine's beneficial effects might be due to its influence on MMP. Human fibroblast-like synoviocytes were used to study the effect of mepacrine on phorbol myristic acetate (PMA) induced activation of MMP. Western blot, reverse transcription-polymerase chain reaction, and electrophoretic mobility shift assay (EMSA) experiments were used to investigate the effect of mepacrine. Mepacrine selectively inhibited MMP in human fibroblast-like synoviocytes. Mepacrine inhibited MMP-1 as well as MMP-9, but had no effect on MMP-3 at the mRNA level. Possible mechanisms to explain these findings were investigated, and it was found that mepacrine had a strikingly different effect on c-Jun, as opposed to c-Fos activation. While mepacrine treatment alone led to increased concentrations of c-Jun within the nuclear compartment, c-Fos translocation into the nucleus was blocked in synoviocytes treated with mepacrine and stimulated with PMA. Accordingly, EMSA showed reduced AP-1 binding in mepacrine treated synoviocytes. These results imply that the observed effects of mepacrine on immediate early genes resulted in reduced AP-1 binding. That ultimately led to a selective suppression of genes that rely on unhindered assembling of these transcription factors for their activation. Selective inhibition of MMP by mepacrine might in part explain the beneficial effects of this drug in the treatment of certain diseases.


The Concept of Patient Participation in Forensic Psychiatric Care: The Patient Perspective.
The importance of patient participation is advocated in medical treatment and nursing care and has been linked to increased quality of care, increased patient satisfaction, and treatment adherence. Still, patients in forensic psychiatric care often report being unhappy with their experienced level of participation. The concept of patient participation is complex and has several definitions, thus it is important to investigate it from different perspectives in different contexts. The aim of this study was to describe patients' perceptions of the concept of patient participation in forensic psychiatric care. A qualitative design with a phenomenographic approach was used, and interviews with 19 participants in a Swedish setting were completed. The participants described the concept of patient participation in forensic psychiatric care as follows: influence, to have good communication and to be involved; confidence, to have mutual trust and to trust the care; and own responsibility, to participate in activities and to take the initiative. On the basis of the results of this study, improved patient participation in forensic psychiatric care may be achieved with active communication, by building up and maintaining trust for professional competence and by encouraging patients' own responsibility. It is important that knowledge about patients' views of the concept of patient participation is included in the planning and improvement of forensic care.


Ontogeny of the mouse vocal fold epithelium.
This investigation provides the first systematic determination of the cellular and molecular progression of vocal fold (VF) epithelium development in a murine model. We define five principal developmental events that constitute the progression from VF initiation in the embryonic anterior foregut tube to fully differentiated and functional adult tissue. These developmental events include (1) the initiation of the larynx and vocal folds with apposition of the lateral walls of the primitive laryngopharynx (embryonic (E) day 10.5); (2) the establishment of the epithelial lamina with fusion of the lateral walls of the primitive laryngopharynx (E11.5); (3) the epithelial lamina recanalization and separation of VFs (E13.5-18.5); (4) the stratification of the vocal folds (E13.5-18.5); and (5) the maturation of vocal fold epithelium (postnatal stages). The illustration of these morphogenetic events is substantiated by dynamic changes in cell proliferation and apoptosis, as well as the expression pattern of key transcription factors, FOXA2, SOX2 and NKX2-1 that specify and pattern the foregut endoderm. Furthermore, we documented the gradual conversion of VF epithelial cells from simple precursors expressing cytokeratins 8 and 18 in the embryo into mature stratified epithelial cells also expressing cytokeratins 5 and 14 in the adult. Interestingly, in the adult, cytokeratins 5 and 14 appear to be expressed in all cell layers in the VF, in contrast to their preferential localization to the basal cell layer in surrounding epithelium. To begin investigating the role of signaling molecules in vocal fold development, we characterized the expression pattern of SHH pathway genes, and how loss of Shh affects vocal fold development in the mutant. This study defines the cellular and molecular context and serves as the necessary foundation for future functional investigations of VF formation.


Characterization of stable human aromatase expressed in E. coli.
Aromatase (P450arom, CYP19) catalyzes the aromatization reaction that converts androgens to estrogens. Although human P450arom has been readily purified from placenta, its hydrophobic properties and instability has hampered detailed characterization. Utilizing a N-terminal replacement (MARQSFGRGKL, derived from CYP2C11), we successfully modified this unstable enzyme into stable forms. Based on a known polymorphism, we created two constructs, NmA264C and NmA264R having cysteine or arginine at position 264. The recombinant P450arom NmA264R was expressed in Escherichia coli (350-400 nmol/L culture) primarily by coexpression with molecular chaperones GroES/GroEL while NmA264C was expressed (240 nmol/L culture) only in the presence of chloramphenicol. Although NmA264C was recovered only in the membrane fraction, approximately 14% of NmA264R was recovered in the cytosolic fraction, suggesting that NmA264R is more hydrophilic than NmA264C. NmA264R was highly purified to the specific content 13.6 nmol P450/mg protein. Purified P450arom NmA264R converted androstenedione to estrone with Vmax 12.4 nmol/(min nmol) and Km) 0.26 microM, and testosterone to estradiol with Vmax 52.2 nmol/(min nmol) and Km 10.9 microM. Because of the increased stability of NmA264R, we could unambiguously determine properties of human P450arom by optical and electron paramagnetic resonance spectroscopy. The purified protein was a typical low-spin form, which was converted to a high-spin form when androstenedione was added. The rhombicity of substrate-bound forms was higher than that reported for other P450s, an interesting characteristic of human P450arom. The highly stable and active P450arom NmA264R sets the stope for detailed structure/function analyses of this important member of the P450 superfamily.


Recombination between viral RNA and transgenic plant transcripts.
Transformed plants expressing the 3' two-thirds of the cowpea chlorotic mottle virus (CCMV) capsid gene were inoculated with a CCMV deletion mutant lacking the 3' one-third of the capsid gene. Although the deletion inoculum replicates in inoculated cells, systemic infections occur only if recombination restores a functional capsid gene. Four of 125 inoculated transgenic plants, representing three different transgenic lines, became systemically infected. Analysis of viral RNA confirmed that RNA recombination had united the transgenic messenger RNA and the challenging virus through aberrant homologous recombination.


Ozone-sensitive arabidopsis rcd1 mutant reveals opposite roles for ethylene and jasmonate signaling pathways in regulating superoxide-dependent cell death.
We have isolated a codominant Arabidopsis mutant, radical-induced cell death1 (rcd1), in which ozone (O(3)) and extracellular superoxide (O(2)(*)-), but not hydrogen peroxide, induce cellular O(2)(*)- accumulation and transient spreading lesions. The cellular O(2)(*)- accumulation is ethylene dependent, occurs ahead of the expanding lesions before visible symptoms appear, and is required for lesion propagation. Exogenous ethylene increased O(2)(*)--dependent cell death, whereas impairment of ethylene perception by norbornadiene in rcd1 or ethylene insensitivity in the ethylene-insensitive mutant ein2 and in the rcd1 ein2 double mutant blocked O(2)(*)- accumulation and lesion propagation. Exogenous methyl jasmonate inhibited propagation of cell death in rcd1. Accordingly, the O(3)-exposed jasmonate-insensitive mutant jar1 displayed spreading cell death and a prolonged O(2)(*)- accumulation pattern. These results suggest that ethylene acts as a promoting factor during the propagation phase of developing oxyradical-dependent lesions, whereas jasmonates have a role in lesion containment. Interaction and balance between these pathways may serve to fine-tune propagation and containment processes, resulting in alternate lesion size and formation kinetics.


Abnormal development of dendritic spines in FMR1 knock-out mice.
Fragile X syndrome is caused by a mutation in the FMR1 gene leading to absence of the fragile X mental retardation protein (FMRP). Reports that patients and adult FMR1 knock-out mice have abnormally long dendritic spines of increased density suggested that the disorder might involve abnormal spine development. Because spine length, density, and motility change dramatically in the first postnatal weeks, we analyzed these properties in mutant mice and littermate controls at 1, 2, and 4 weeks of age. To label neurons, a viral vector carrying the enhanced green fluorescent protein gene was injected into the barrel cortex. Layer V neurons were imaged on a two-photon laser scanning microscope in fixed tissue sections. Analysis of >16,000 spines showed clear developmental patterns. Between 1 and 4 weeks of age, spine density increased 2.5-fold, and mean spine length decreased by 17% in normal animals. Early during cortical synaptogenesis, pyramidal cells in mutant mice had longer spines than controls. At 1 week, spine length was 28% greater in mutants than in controls. At 2 weeks, this difference was 10%, and at 4 weeks only 3%. Similarly, spine density was 33% greater in mutants than in controls at 1 week of age. At 2 or 4 weeks of age, differences were not detectable. The spine abnormality was not detected in neocortical organotypic cultures. The transient nature of the spine abnormality in the intact animal suggests that FMRP might play a role in the normal process of dendritic spine growth in coordination with the experience-dependent development of cortical circuits.


Molecular detection of Peronospora variabilis in quinoa seed and phylogeny of the quinoa downy mildew pathogen in South America and the United States.
Quinoa (Chenopodium quinoa) is an important export of the Andean region, and its key disease is quinoa downy mildew, caused by Peronospora variabilis. P. variabilis oospores can be seedborne and rapid methods to detect seedborne P. variabilis have not been developed. In this research, a polymerase chain reaction (PCR)-based detection method was developed to detect seedborne P. variabilis and a sequencing-based method was used to validate the PCR-based method. P. variabilis was detected in 31 of 33 quinoa seed lots using the PCR-based method and in 32 of 33 quinoa seed lots using the sequencing-based method. Thirty-one of the quinoa seed lots tested in this study were sold for human consumption, with seed originating from six different countries. Internal transcribed spacer (ITS) and cytochrome c oxidase subunit 2 (COX2) phylogenies were examined to determine whether geographical differences occurred in P. variabilis populations originating from Ecuador, Bolivia, and the United States. No geographical differences were observed in the ITS-derived phylogeny but the COX2 phylogeny indicated that geographical differences existed between U.S. and South American samples. Both ITS and COX2 phylogenies supported the existence of a Peronospora sp., distinct from P. variabilis, that causes systemic-like downy mildew symptoms on quinoa in Ecuador. The results of these studies allow for a better understanding of P. variabilis populations in South America and identified a new causal agent for quinoa downy mildew. The PCR-based seed detection method allows for the development of P. variabilis-free quinoa seed, which may prove important for management of quinoa downy mildew.


Sensitivity to bee and wasp venoms: association with specific IgE responses to the bee and wasp venom and HLA DRB1 and DPB1.
Stings from bees and wasps can cause systemic reactions which can be fatal in some individuals. In these venom-sensitive patients, specific IgE to the venom is produced and is considered to participate in the adverse reactions. This immune response requires antigen presentation by human leucocyte antigens (HLA) class II molecules, which includes DR and DP, which are present on antigen presenting cells. To test for associations between HLA class II DRB1 and DPB1 alleles and life-threatening sensitivity to both bee and wasp venoms. To establish further whether any associations are independent of the atopy phenotype. A total of 33 bee- and 44 wasp-venom-sensitive patients was studied. DRB1 genotypes were determined by single stranded oligonucleotide (SSO) probing of PCR products, and DPB1 genotypes by amplified fragment length polymorphism (AFLP) analysis. Total and specific IgE were measured using the Pharmacia Immunocap, FEIA. Patients with specific IgE to the venom antigens only were termed monosensitive and those with additional specific IgE to HDM and/or GP were termed polysensitive. Allele frequencies were compared to an unrelated control population. The 33 bee-sensitive patients had a greater prevalence of DRB1*07 alleles than the control subjects, 26% vs 14%, with an odds ratio (OR) of 2.1 (95% CI, 1.2-3.7, P = 0.015, corrected for multiple comparisons, Pc = ns). This association was confined to the 15 monosensitive bee patients, who had a 43% DRB1*07 allele frequency when compared with 11% in the 18 polysensitive bee patients, OR 6.1 (95% CI, 1.73-22, P = 0.004, Pc = 0.05), and when compared with a control group of non-venom subjects, 43% vs 16%, OR 3.9 (95% CI, 1.72-9.0, P = 0.002, Pc = 0.02). The 44 wasp-sensitive patients had an increase in the DRB1*11 allele when compared with the control subjects, 13% vs 6%, with an OR 2.2 (95% CI, 1.0-4.6, P = 0.04, Pc = NS), and a decreased prevalence of DRB1*04 alleles, 10% vs 19%, with an OR 0.33 (95% CI, 0.24-0.99, P = 0.04, Pc = NS), but these were not significant when multiple comparisons were taken into account. The DPB1 alleles were not significantly different between the venom sensitive patients and the controls. Patients monosensitive to bee venom had a significantly greater prevalence of DRB1*07 alleles than the non-venom, control population suggesting that IgE responses in these patients may, in part be controlled by immune response HLA class II genes. These results are also suggestive of wasp-sensitive patients having a higher prevalence of DRB1*11 and a lower prevalence of DRB1*04 than the control population.


Evaluation of cannabinoid receptor binding and in vivo activities for anandamide analogs.
Recent evidence implicates anandamide as the endogenous ligand for the cannabinoid receptor. One purpose of this study was to determine the structural requirements for anandamide's receptor interaction and the influence of phenylmethylsulfonyl fluoride (PMSF), an enzyme inhibitor, on receptor affinity. A second objective was evaluation of the correlation between affinities of the analogs and in vivo pharmacological activities. The ability of anandamide and analogs to displace [3H]CP-55,940 ([3](-)-3-[2-hydroxyl-4-(1,1-dimethylheptyl)phenyl]-4-[3- hydroxylpropyl]cyclohexan-1-ol) was determined by a filtration assay. Displacement curves for anandamide in the presence of PMSF produced a Ki of 89 +/- 10 nM; without PMSF the Ki increased to 5400 +/- 1600 nM. Anandamide analogs were evaluated for their ability to produce antinociception and hypomotility. The levels of saturation of the anandamide structure were critical to receptor affinity and in vivo potency, with complete saturation and hydroxyl substitution with a fluorine moiety resulting in a compound with increased potency in the spontaneous activity and antinociception assays. Substitution of the hydroxyl with a fluorine atom increased affinity only in the presence of PMSF and reduced potency in the antinociception assay. Ethanolamide substitution with bromobenzenesulfonamide produced an inactive compound in all assays. Increasing the length of the N-substituent by one or two carbons decreased receptor binding affinity and potency in the tail-flick assay only. Certain structural modifications, such as methylations, allowed the analogs to retain affinity without the addition of PMSF. Linear correlation between the behavioral and binding assays were performed, and the greatest correlation was obtained with compounds that were either very potent or inactive.


Myofascial trigger points, neck mobility, and forward head posture in episodic tension-type headache.
To assess the differences in the presence of trigger points (TrPs) in head and neck muscles, forward head posture (FHP) and neck mobility between episodic tension-type headache (ETTH) subjects and healthy controls. In addition, we assess the relationship between these muscle TrPs, FHP, neck mobility, and several clinical variables concerning the intensity and the temporal profile of headache. TTH is a headache in which musculoskeletal disorders of the craniocervical region might play an important role in its pathogenesis. Design.-A blinded, controlled pilot study. Fifteen ETTH subjects and 15 matched controls without headache were studied. TrPs in both upper trapezius, both sternocleidomastoids, and both temporalis muscles were identified according to Simons and Gerwin diagnostic criteria (tenderness in a hypersensible spot within a palpable taut band, local twitch response elicited by snapping palpation, and elicited referred pain with palpation). Side-view pictures of each subject were taken in both sitting and standing positions, in order to assess FHP by measuring the craniovertebral angle. A cervical goniometer was employed to measure neck mobility. All measures were taken by a blinded assessor. A headache diary was kept for 4 weeks in order to assess headache intensity, frequency, and duration. The mean number of TrPs for each ETTH subject was 3.7 (SD: 1.3), of which 1.9 (SD: 0.9) were active, and 1.8 (SD: 0.9) were latent. Control subjects only had latent TrPs (mean: 1.5; SD: 1). TrP occurrence between the 2 groups was significantly different for active TrPs (P < .001), but not for latent TrPs (P > .05). Differences in the distribution of TrPs were significant for the right upper trapezius muscles (P= .04), the left sternocleidomastoid (P= .03), and both temporalis muscles (P < .001). Within the ETTH group, headache intensity, frequency, and duration outcomes did not differ depending on TrP activity, whether the TrP was active or latent. The craniovertebral angle was smaller, ie, there was a greater FHP, in ETTH patients than in healthy controls for both sitting and standing positions (P < .05). ETTH subjects with active TrPs in the analyzed muscles had a greater FHP than those with latent TrPs in both sitting and standing positions, though differences were only significant for certain muscles. Finally, ETTH patients also showed lesser neck mobility than healthy controls in the total range of motion as well as in half-cycles (except for cervical extension), although neck mobility did not seem to influence headache parameters. Active TrPs in the upper trapezius, sternocleidomastoid, and temporalis muscles were more common in ETTH subjects than in healthy controls, although TrP activity was not related to any clinical variable concerning the intensity and the temporal profile of headache. ETTH patients showed greater FHP and lesser neck mobility than healthy controls, although both disorders were not correlated with headache parameters.


Prevalence of hepatitis B and C and HIV infections among problem drug users in Luxembourg: self-report versus serological evidence.
To determine the seroprevalence of hepatitis B (HBV), hepatitis C (HCV) and HIV infections in problem drug users (PDU) in Luxembourg. To measure the validity of self-reported test results provided by study participants as well as obtained through the national drug-monitoring system (RELIS). In a cross-sectional multisite study, data were collected by voluntary, anonymous and assisted questionnaires and serological detection of antibodies and antigens. Out of 1169 contacts, 397 participants were recruited within in and out-of-treatment settings (84.2% injecting drug users; IDU). The prevalence of antibodies to HIV was 8/272 (2.9%; 95% CI 0.9% to 4.9%), to HCV 245/343 (71.4%; 66.6% to 76.2%), and 67/310 (21.6%; 17.1% to 26.2%) to total HBV antibodies and surface antigen (for IDU 5/202, 218/268 and 59/239, respectively). Specificity of study self-reports was very high for HBV and perfect for HCV and HIV. Sensitivity was 0.224, 0.798 and 0.800, respectively. Kappa scores provided degrees of agreement between serological tests and study self-reports of 0.89 for HIV, 0.65 for HCV and 0.25 for HBV. In contrast to simultaneous cross-sectional self-reports, secondary self-reported data (RELIS) showed high agreement for HIV and HBV infections and provided a good proxy for estimation of HCV seroprevalence. HIV testing routines in PDU should be completed at least by HBV and HCV detection given the poor validity of cross-sectional self-reports on hepatitis infections. HIV and hepatitis prevalence estimations in PDU gain by relying on multisite/setting data collection. Research should further investigate the validity of HIV and hepatitis self-reports from routine drug-monitoring systems versus cross-sectional surveys.


Nonspherical double emulsions with multiple distinct cores enveloped by ultrathin shells.
Microfluidics has provided means to control emulsification, enabling the production of highly monodisperse double-emulsion drops; they have served as useful templates for production of microcapsules. To provide new opportunities for double-emulsion templates, here, we report a new design of capillary microfluidic devices that create nonspherical double-emulsion drops with multiple distinct cores covered by ultrathin middle layer. To accomplish this, we parallelize capillary channels, each of which has a biphasic flow in a form of core-sheath stream; this is achieved by preferential wetting of oil to the hydrophobic wall. These core-sheath streams from the parallelized channels are concurrently emulsified into continuous phase, making paired double-emulsion drops composed of multiple cores and very thin middle shell. This microfluidic approach provides high degree of controllability and flexibility on size, shape, number, and composition of double-emulsion drops. Such double-emulsion drops are useful as templates to produce microcapsules with multicompartments which can encapsulate and deliver multiple distinct components, while avoiding their cross-contamination. In addition, nonspherical envelope exerts strong capillary force, leading to preferential coalescence between innermost drops; this is potentially useful for nanoliter-scale reactions and encapsulations of the reaction products.


Use of a microchip device coupled with mass spectrometry for ligand screening of a multi-protein target.
Nanoflow electrospray mass spectrometry has been applied previously to investigate noncovalent protein-protein and protein-ligand interactions. Here we evaluate a commercial microchip device for this application. We show that the microchip can be used to obtain mass spectra of the noncovalent tetramer transthyretin. The device showed a 10-fold increase in signal stability compared with a nanoflow capillary and a high level of nozzle-to-nozzle reproducibility. Binding of the natural ligand thyroxine was clearly observed, and a range of small molecules proposed as inhibitors of transthyretin amyloidosis were shown to be effective in stabilizing the tetramer. We propose that measuring the ability of small molecules to stabilize protein complexes using this automated microchip technology will enable high-throughput screening of multi-protein complexes by mass spectrometry.


Effects of a porphyrazine derivative on some developmental characteristics of Drosophila melanogaster.
In this study, the effects of a porphyrazine derivative, [octakis (2-trimethylammoniumethyl thio) porphyrazinatocobalt] octaiodide (CoPzq), were explored on the developmental characteristics, sex ratio and egg fertility of Drosophila melanogaster. CoPzq was preferred in the study due to its probability to be used as a chemotherapeutic agent in medicine. As a result of the application of CoPzq, a phenotypic abnormality was revealed within the individuals of F1 and F2 generation of Drosophila. Although individuals of F2 generation did not directly come into contact with the chemical, the rate of abnormality is high. An increase was observed within both generations in terms of the egg fertility. Besides, a deviation emerged in the sex-ratio in some concentrations.


Does maternal exposure to artificial food coloring additives increase oxidative stress in the skin of rats?
Glutathione-S-transferase (GST) and cytochrome P450 family 1 subfamily A polypeptide 1 (CYP1A1) metabolize and detoxify carcinogens, drugs, environmental pollutants, and reactive oxygen species. Changes of GST expression in tissues and gene mutations have been reported in association with many neoplastic skin diseases and dermatoses. Widely used artificial food coloring additives (AFCAs) also reported to effect primarily behavioral and cognitive function and cause neoplastic diseases and several inflammatory skin diseases. We aimed to identify the changes in expression of GSTs, CYP1A1, and vascular endothelial growth factor (VEGF) in rat skin which were maternally exposed AFCAs. A rat model was designed to evaluate the effects of maternal exposure of AFCAs on skin in rats. "No observable adverse effect levels" of commonly used AFCAs as a mixture were given to female rats before and during gestation. Immunohistochemical expression of GSTs, CYP1A1, and VEGF was evaluated in their offspring. CYP1A1, glutathione S-transferase pi (GSTP), glutathione S-transferase alpha (GSTA), glutathione S-transferase mu (GSTM), glutathione S-transferase theta (GSTT), and VEGF were expressed by epidermal keratinocytes, dermal fibroblasts, sebaceous glands, hair follicle, and subcutaneous striated muscle in the normal skin. CYP1A1, GSTA, and GSTT were expressed at all microanatomical sites of skin in varying degrees. The expressions of CYP1A1, GSTA, GSTT, and VEGF were decreased significantly, while GSTM expression on sebaceous gland and hair follicle was increased. Maternal exposure of AFCAs apparently effects expression of the CYP1A1, GSTs, and VEGF in the skin. This prominent change of expressions might play role in neoplastic and nonneoplastic skin diseases.


ESNM/ANMS consensus paper: Diagnosis and management of refractory gastro-esophageal reflux disease.
Up to 40% of patients with symptoms suspicious of gastroesophageal reflux disease (GERD) do not respond completely to proton pump inhibitor (PPI) therapy. The term "refractory GERD" has been used loosely in the literature. A distinction should be made between refractory symptoms (ie, symptoms may or may not be GERD-related), refractory GERD symptoms (ie, persisting symptoms in patients with proven GERD, regardless of relationship to ongoing reflux), and refractory GERD (ie, objective evidence of GERD despite adequate medical management). The present ESNM/ANMS consensus paper proposes use the term "refractory GERD symptoms" only in patients with persisting symptoms and previously proven GERD by either endoscopy or esophageal pH monitoring. Even in this context, symptoms may or may not be reflux related. Objective evaluation, including endoscopy and esophageal physiologic testing, is requisite to provide insights into mechanisms of symptom generation and evidence of true refractory GERD. Some patients may have true ongoing refractory acid or weakly acidic reflux despite PPIs, while others have no evidence of ongoing reflux, and yet others have functional esophageal disorders (overlapping with proven GERD confirmed off therapy). In this context, attention should also be paid to supragastric belching and rumination syndrome, which may be important contributors to refractory symptoms.


A hierarchical classification of polysaccharide lyases for glycogenomics.
Carbohydrate-active enzymes face huge substrate diversity in a highly selective manner using only a limited number of available folds. They are therefore subjected to multiple divergent and convergent evolutionary events. This and their frequent modularity render their functional annotation in genomes difficult in a number of cases. In the present paper, a classification of polysaccharide lyases (the enzymes that cleave polysaccharides using an elimination instead of a hydrolytic mechanism) is shown thoroughly for the first time. Based on the analysis of a large panel of experimentally characterized polysaccharide lyases, we examined the correlation of various enzyme properties with the three levels of the classification: fold, family and subfamily. The resulting hierarchical classification, which should help annotate relevant genes in genomic efforts, is available and constantly updated at the Carbohydrate-Active Enzymes Database (http://www.cazy.org).


Anticoagulation Knowledge Tool (AKT): Further evidence of validity in the Italian population.
Oral Anticoagulation therapy (OAC) is highly effective in the management of thromboembolic disorders. An adequate level of knowledge is important for self-management and optimizing clinical outcomes. The Anticoagulation Knowledge Tool (AKT) was developed to assess OAC knowledge and caters for both patients prescribed direct oral anticoagulants or vitamin K antagonist (VKA). However, evidence regarding its psychometric proprieties, validity and reliability are unavailable in non-English speaking settings. For this reason, the aim of this study is to provide further evidence of validity for AKT and also developing an Italian AKT version (I-AKT) supported by evidence of validity and reliability. A multiphase study was conducted which included the following: cultural and linguistic validity; i.e. content validity; construct validity; reliability assessment. The Construct validity was performed using the contrasted group approach using three groups comprised of health care providers, patients and the general public. Furthermore, Exploratory Structural Equation Modelling (ESEM) was performed to confirm the mono-dimensional structure of the items in the AKT. In construct validity phase 334 participants were enrolled. One-way ANOVA and post hoc analysis test demonstrated significant differences between the means knowledge scores of the three groups: 30.42±3.04 vs 23.45± 4.57 vs14.32±6.07 (Statistic F = 266.83; p < .001). ESEM analysis demonstrates the I-AKT mono-dimensionally structure with an explained variance of 56.42%. The scale also showed both good internal consistency reliability (Cronbach's α = 0.896) and test-retest reliability (r = 0.855). This study developed and validated I-AKT with supporting evidence for validity and reliability. The study also confirms the mono-dimensional of the items in the AKT. This suggest that the instrument can be useful in non-English setting for knowledge assessment and in potentially developing patient education materials.


Bolus matters: the influence of food oral breakdown on dynamic texture perception.
This review article focuses on design of food structure, characterisation of oral processing by boli characterisation and dynamic texture perception. Knowledge of the food properties governing bolus formation and bolus properties determining temporal changes in texture perception is of major importance. Such knowledge allows academia to better understand the mechanisms underlying texture perception and food industry to improve product texture. For instance, such knowledge can be used for developing foods with desired texture perception that fit in a healthy diet or that are customized to specific consumer groups. The end point of oral processing is the formation of a safe-to-swallow bolus. The transitions of solid and soft solid foods into bolus are accompanied by tremendous modifications of food properties. The review discusses dynamic changes in bolus properties resulting in dynamic changes of texture perception during oral processing. Studies monitoring chewing behaviour are discussed to complement the relationships between bolus properties and dynamic texture perception. We conclude that texture perception evolves over mastication time and depends on food properties, such as mechanical properties, mainly in the beginning of oral processing. Towards the middle and end of oral processing, bolus properties depend on food properties and explain texture perception better than food properties.


Islets of Langerhans: dye coupling among immunocytochemically distinct cell types.
Cells in isolated rat islets of Langerhans were microinjected with Lucifer Yellow CH in a medium containing 16.7 millimolar glucose. Dye was rapidly transferred from the injected cell to neighboring islet cells without specificity with regard to the immunocytochemical identity of either the donor or the recipient cells. The transfer of dye between the islet cells (types A, B, and D) demonstrates homologous and heterologous cell coupling in a system where the normal proportions and relationships of the cell types are maintained.


Using ChIP-seq technology to generate high-resolution profiles of histone modifications.
The dynamic modification of DNA and histones plays a key role in transcriptional regulation through -altering the packaging of DNA and modifying the nucleosome surface. These chromatin states, also referred to as the epigenome, are distinctive for different tissues, developmental stages, and disease states and can also be altered by environmental influences. New technologies allow the genome-wide visualization of the information encoded in the epigenome. For example, the chromatin immunoprecipitation (ChIP) assay allows investigators to characterize DNA-protein interactions in vivo. ChIP followed by hybridization to microarrays (ChIP-chip) or by high-throughput sequencing (ChIP-seq) are both powerful tools to identify genome-wide profiles of transcription factors, histone modifications, DNA methylation, and nucleosome positioning. ChIP-seq technology, which can now interrogate the entire human genome at high resolution with only one lane of sequencing, has recently surpassed ChIP-chip technology for epigenomic analyses. Importantly, for the study of primary cells and tissues, epigenetic profiles can be generated using as little as 1 μg of chromatin. In this chapter, we describe in detail the steps involved in performing ChIP assays (with a focus on characterizing histone modifications in primary cells) either manually or using the IP-Star ChIP robot, followed by a detailed protocol to prepare successful libraries for Illumina sequencing. Critical quality control checkpoints are discussed. Although not a focus of this chapter, we also point the reader to several methods by which massive ChIP-seq data sets can be analyzed to extract the tremendous information contained within.


Placental inflammation is not increased in inflammatory bowel disease.
Women with inflammatory bowel disease (IBD) are at increased risk for adverse birth outcomes such as preterm delivery and small for gestational age (SGA) infants. Most recognized cases of fetal growth restriction in singleton pregnancies have underlying placental causes. However, studies in IBD examining poor birth outcomes have focused on maternal factors. We examined whether women with IBD have a higher rate of placental inflammation than non-IBD controls. Between 2008 and 2011, the placental tissue of 7 ulcerative colitis, 5 Crohn's disease, and 2 IBD-unclassified subjects enrolled in the Pregnancy in Inflammatory Bowel Disease and Neonatal Outcome (PIANO) registry were evaluated for villitis, deciduitis, and chorioamnionitis with/without a fetal inflammatory response. The history and birth outcomes of all IBD subjects were reviewed and matched to 26 non-IBD controls by gestational age at delivery. Of women with IBD, 29% delivered preterm infants and 21% delivered SGA infants. Half of the IBD patients had mild-moderate disease flares during pregnancy. Five (36%) patients required corticosteroids, 2 (14%) were maintained on an immunomodulator, and 3 (21%) others received tumor necrosis factor-alpha inhibitors during their pregnancy. Chorioamnionitis was the only identified placental pathology present in the placentas reviewed, occurring less frequently in cases compared to controls (7% vs. 27%, P=0.32). Placental inflammatory activation does not appear to be responsible for the increase in adverse birth outcome in women with IBD. Further studies are necessary to validate these findings in IBD to explain poor birth outcomes.


Resistance and Resilience of Sulfidogenic Communities in the Face of the Specific Inhibitor Perchlorate.
Hydrogen sulfide is a toxic and corrosive gas, produced by the activity of sulfate-reducing microorganisms (SRM). Owing to the environmental, economic and human-health consequences of sulfide, there is interest in developing specific inhibitors of SRM. Recent studies have identified perchlorate as a promising emerging inhibitor. The aim of this work is to quantitatively dissect the inhibitory dynamics of perchlorate. Sulfidogenic mixed continuous-flow systems were treated with perchlorate. SRM number, sulfide production and community structure were monitored pre-, during and post-treatment. The data generated was compared to a simple mathematical model, where SRM growth slows as a result of inhibition. The experimental data supports the interpretation that perchlorate largely acts to suppress SRM growth rates, rendering planktonic SRM increasingly susceptible to wash-out. Surface-attachment was identified as an important parameter preventing SRM wash-out and thus governing inhibitory dynamics. Our study confirmed the lesser depletion of surface-attached SRM as compared to planktonic SRM during perchlorate treatment. Indirect effects of perchlorate (bio-competitive exclusion of SRM by dissimilatory perchlorate-reducing bacteria, DPRB) were also assayed by amending reactors with DPRB. Indeed, low concentrations of perchlorate coupled with DRPB amendment can drive sulfide concentrations to zero. Further, inhibition in a complex community was compared to that in a pure culture, highlighting similarities and differences between the two scenarios. Finally, we quantified susceptibility to perchlorate across SRM in various culture conditions, showing that prediction of complex behavior in continuous systems from batch results is possible. This study thus provides an overview of the sensitivity of sulfidogenic communities to perchlorate, as well as mechanisms underlying these patterns.


Quantum and classical dynamics of a Bose-Einstein condensate in a large-period optical lattice.
We experimentally investigate diffraction of a 87Rb Bose-Einstein condensate from a one-dimensional optical lattice. We use a range of lattice periods and timescales, including those beyond the Raman-Nath limit. We compare the results to numerical solutions of the Gross-Pitaevskii equation and classical calculations, with quantitative and qualitative agreement, respectively. The classical calculations predict that the envelope of the time-evolving diffraction pattern is shaped by caustics: singularities in the phase-space density of classical trajectories. This behavior becomes increasingly clear as the lattice period grows.


Progression-Free Survival as a Surrogate End Point for Overall Survival in First-Line Diffuse Large B-Cell Lymphoma: An Individual Patient-Level Analysis of Multiple Randomized Trials (SEAL).
Purpose Overall survival (OS) is the definitive and best-established primary efficacy end point to evaluate diffuse large B-cell lymphoma (DLBCL) therapies, but it requires prolonged follow-up. An earlier end point assessed post-treatment would expedite clinical trial conduct and accelerate patient access to effective new therapies. Our objective was to formally evaluate progression-free survival (PFS) and PFS at 24 months (PFS24) as surrogate end points for OS in first-line DLBCL. Patients and Methods Individual patient data were analyzed from 7,507 patients from 13 multicenter randomized controlled trials of active treatment in previously untreated DLBCL, published after 2002, with sufficient PFS data to predict treatment effects on OS. Trial-level surrogacy examining the correlation of treatment effect estimates of PFS/PFS24 and OS was evaluated using both linear regression ( R2WLS) and Copula bivariable ( R2Copula) models. Prespecified criteria for surrogacy required either R2WLS or R2Copula ≥ 0.80 and neither < 0.7, with lower-bound 95% CI > 0.60. Results Trial-level surrogacy for PFS was strong ( R2WLS = 0.83; R2Copula = 0.85) and met the predefined criteria for surrogacy. At the patient level, PFS strongly correlated with OS. The surrogate threshold effect had a hazard ratio of 0.89. Surrogacy was consistent across comparisons with or without rituximab and with rituximab maintenance trials. Trial-level surrogacy for PFS24 was relatively strong ( R2WLS = 0.77; R2Copula = 0.78) but did not meet prespecified criteria. At the patient level, PFS24 significantly correlated with OS. The surrogate threshold effect had an odds ratio of 1.51. Conclusion This large pooled analysis of individual patient data supports PFS as a surrogate end point for OS in future randomized controlled trials evaluating chemoimmunotherapy in DLBCL. Use of this end point may expedite therapeutic development with the intent of bringing novel therapies to this patient population years before OS results are mature.


Analytical calculation of sensing parameters on carbon nanotube based gas sensors.
Carbon Nanotubes (CNTs) are generally nano-scale tubes comprising a network of carbon atoms in a cylindrical setting that compared with silicon counterparts present outstanding characteristics such as high mechanical strength, high sensing capability and large surface-to-volume ratio. These characteristics, in addition to the fact that CNTs experience changes in their electrical conductance when exposed to different gases, make them appropriate candidates for use in sensing/measuring applications such as gas detection devices. In this research, a model for a Field Effect Transistor (FET)-based structure has been developed as a platform for a gas detection sensor in which the CNT conductance change resulting from the chemical reaction between NH3 and CNT has been employed to model the sensing mechanism with proposed sensing parameters. The research implements the same FET-based structure as in the work of Peng et al. on nanotube-based NH3 gas detection. With respect to this conductance change, the I-V characteristic of the CNT is investigated. Finally, a comparative study shows satisfactory agreement between the proposed model and the experimental data from the mentioned research.


Sensitivity and specificity of urinary neutrophil gelatinase-associated lipocalin and kidney injury molecule-1 for the diagnosis of renal cell carcinoma.
Neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) are urinary biomarkers of diagnostic relevance in a wide variety of acute and chronic kidney diseases. Their diagnostic sensitivity and specificity for kidney cancer are largely unknown and therefore the subject of this investigation. A prospective cohort study was performed to evaluate urine biomarkers for clear-cell and papillary subtypes of renal cancer (67 patients undergoing nephrectomy) and 55 control patients undergoing non-kidney surgery. Urinary KIM-1 and NGAL concentrations were determined by sensitive and specific ELISAs. In renal cancer patients, median NGAL excretion was 0.52 (1st to 3rd quartiles: 0.28-0.82) ng/mg urinary creatinine (U(Cr)) before nephrectomy compared to 0.15 (0.04-0.31) ng/mg U(Cr) in controls (p < 0.001), and there was a modest decrease of 30% after nephrectomy (p < 0.008). NGAL was not correlated to tumor size (r = 0.19, p = 0.27) or stage. Before nephrectomy, KIM-1 excretion was 0.68 (0.40-1.12) ng/mg U(Cr) compared to 0.03 (0.01-0.06) in controls (p < 0.001). There was a linear correlation between KIM-1 excretion before nephrectomy and tumor size (Spearman's r = 0.66, p < 0.001), tumor stage, and a 50% decrease in median KIM-1 concentration 1 month following tumor excision (p < 0.01). Biomarker concentration ranges for renal cancer patients and controls overlapped substantially for NGAL but not KIM-1. NGAL is not a sensitive or specific urinary biomarker of kidney cancer. Although KIM-1 had diagnostic sensitivity for kidney cancer, it is well known to reflect many types of kidney injuries, thus limiting its specificity as a diagnostic biomarker for renal cancer.


Private sector malaria RDT initiative in Nigeria: lessons from an end-of-project stakeholder engagement meeting.
The malaria rapid diagnosis testing (RDT) landscape is rapidly evolving in health care delivery in Nigeria with many stakeholders playing or having potential for critical roles. A recent UNITAID grant supported a pilot project on the deployment of quality-assured RDTs among formal and informal private service outlets in three states in Nigeria. This paper describes findings from a series of stakeholder engagement meetings held at the conclusion of the project. The agreed meeting structure was a combination of plenary presentations, structured facilitated discussions, and nominal group techniques to achieve consensus. Rapporteurs recorded the meeting proceeding and summaries of the major areas of discussion and consensus points through a retrospective thematic analysis of the submitted meeting reports. Key findings indicate that private providers were confident in the use of RDTs for malaria diagnosis and believed it has improved the quality of their services. However, concerns were raised about continued access to quality-assured RDT kits. Going forward, stakeholders recommended increasing client-driven demand, and continuous training and supervision of providers through integration with existing monitoring and supervision mechanisms.


Coffee consumption and risk of hypertension in the SUN Project.
Evidence on coffee consumption and its association with the incidence of hypertension is still inconsistent. The aim of this study was to examine the association of regular or decaffeinated coffee consumption with the risk of developing hypertension in a middle-aged Mediterranean cohort. The SUN Project is a prospective open cohort with more than 22,500 Spanish university graduates. For the present study, we analyzed data from 13,374 participants initially free of hypertension (mean follow-up 9.1 years). The consumption of regular and decaffeinated coffee was obtained at baseline using a previously validated semi-quantitative food frequency questionnaire. Validated, self-reported medical diagnoses of hypertension were collected biennially. We used Cox regression models to estimate hazard ratios (HR) and 95% confidence intervals (95% CI) for incident hypertension according to baseline coffee consumption. We assessed the interaction with sex and baseline adherence to the Mediterranean diet. Among 121,397 person-years of follow-up, a total of 1757 participants developed hypertension. Overall, coffee consumption -either caffeinated or decaffeinated- was not significantly associated with the risk of hypertension. Only among women, higher consumption of regular coffee was associated with a 26% lower risk of hypertension (>=2 cups/d vs. never/seldom, 95% CI 9%-39%; p for interaction: 0.0236). Women with a low baseline adherence to the Mediterranean diet showed the strongest risk reduction (HR ≥ 2 cups/d vs. never/seldom 0.58, 95% CI (0.41-0.82) p for interaction = 0.0452). In the SUN project we found an inverse association between regular coffee consumption and the risk of hypertension in women, which was strongest among women with a suboptimal food pattern (low adherence to the Mediterranean diet).


Regulation by vitamin D metabolites of parathyroid hormone gene transcription in vivo in the rat.
In vitro 1,25-dihydroxycholecalciferol (1,25(OH)2D3) decreased levels of preproparathyroid(preproPTH) hormone mRNA. We have now pursued these studies in vivo in the rat. Rats were administered vitamin D metabolites i.p. and the levels of preproPTH mRNA were determined in excised parathyroid-thyroid glands by blot hybridization. PreproPTH mRNA levels were less than 4% of basal at 48 h after 100 pmol 1,25(OH)2D3, with no increase in serum calcium. Gel blots showed that 1,25(OH)2D3 decreased preproPTH mRNA levels without any change in its size (833 basepair). Microdissected parathyroids after 1,25(OH)2D3 (100 pmol) showed mRNA levels for preproPTH were 40 +/- 8% of controls, but for beta-actin were 100% of controls. The relative potencies of vitamin D metabolites were: 1,25(OH)2D3 greater than 24,25(OH)2D3 greater than 25(OH)D3 greater than vitamin D3. In vitro nuclear transcription showed that 1,25(OH)2D3-treated (100 pmol) rats' PTH transcription was 10% of control, while beta-actin was 100%. These results show that 1,25(OH)2D3 regulates PTH gene transcription. PTH stimulates 1,25(OH)2D3 synthesis, which then inhibits PTH synthesis, thus completing an endocrinological feedback loop.


Signal categorization by foraging animals depends on ecological diversity.
Warning signals displayed by defended prey are mimicked by both mutualistic (Müllerian) and parasitic (Batesian) species. Yet mimicry is often imperfect: why does selection not improve mimicry? Predators create selection on warning signals, so predator psychology is crucial to understanding mimicry. We conducted experiments where humans acted as predators in a virtual ecosystem to ask how prey diversity affects the way that predators categorize prey phenotypes as profitable or unprofitable. The phenotypic diversity of prey communities strongly affected predator categorization. Higher diversity increased the likelihood that predators would use a 'key' trait to form broad categories, even if it meant committing errors. Broad categorization favors the evolution of mimicry. Both species richness and evenness contributed significantly to this effect. This lets us view the behavioral and evolutionary processes leading to mimicry in light of classical community ecology. Broad categorization by receivers is also likely to affect other forms of signaling.


Malignant Peritoneal Mesothelioma Masquerades as Peritoneal Metastasis on (18)F-FDG PET/CT Scans; a Rare Diagnosis that Should Not Be Missed.
Malignant peritoneal mesothelioma (MPM) is a rare but fatal tumor. The clinical presentations and imaging findings are nonspecific and resemble various diseases, including peritoneal metastasis. Imaging findings of MPH on (18)F-(18)F-fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET)/computed tomography (CT) are diverse and not well described. We report the two cases of biopsy-proven MPH using (18)F-FDG PET/CT. In our cases, interesting disease patterns-including MPH arising from visceral peritoneal lining of kidney that suffer from polycystic disease and from the parietal peritoneum beneath the appendectomy scar-were presented. One case showed classical metastases localized within the abdominal cavity; while the other case exhibited the rare pattern of extensive multi-organ metastases. By knowing the possible variations and diagnostic pitfalls of (18)F-FDG PET/CT findings in MPM, more accurate interpretation of such mysterious cancer is attainable.


Parkinson disease mutant E46K enhances α-synuclein phosphorylation in mammalian cell lines, in yeast, and in vivo.
Although α-synuclein (α-syn) phosphorylation has been considered as a hallmark of sporadic and familial Parkinson disease (PD), little is known about the effect of PD-linked mutations on α-syn phosphorylation. In this study, we investigated the effects of the A30P, E46K, and A53T PD-linked mutations on α-syn phosphorylation at residues Ser-87 and Ser-129. Although the A30P and A53T mutants slightly affected Ser(P)-129 levels compared with WT α-syn, the E46K mutation significantly enhanced Ser-129 phosphorylation in yeast and mammalian cell lines. This effect was not due to the E46K mutant being a better kinase substrate nor due to alterations in endogenous kinase levels, but was mostly linked with enhanced nuclear and endoplasmic reticulum accumulation. Importantly, lentivirus-mediated overexpression in mice also showed enhanced Ser-129 phosphorylation of the E46K mutant compared to WT α-syn, thus providing in vivo validation of our findings. Altogether, our findings suggest that the different PD-linked mutations may contribute to PD pathogenesis via different mechanisms.


Postural synergies and their development.
The recent developments of a particular approach to analyzing motor synergies based on the principle of motor abundance has allowed a quantitative assessment of multi-effector coordination in motor tasks involving anticipatory adjustments to self-triggered postural perturbations and in voluntary postural sway. This approach, the uncontrolled manifold (UCM) hypothesis, is based on an assumption that the central nervous system organizes covariation of elemental variables to stabilize important performance variables in a task-specific manner. In particular, this approach has been used to demonstrate and to assess the emergence of synergies and their modification with motor practice in typical persons and persons with Down syndrome. The framework of the UCM hypothesis allows the formulation of testable hypotheses with respect to developing postural synergies in typically and atypically developing persons.


The effect of eating speed at breakfast on appetite hormone responses and daily food consumption.
The effect of eating speed at a meal on appetite gut hormone responses and future food consumption is not clear. This study examined the effect of eating speed at breakfast on postprandial gut hormone responses, subjective appetite, and daily food consumption. Twenty-five participants [68% men; age, 25.9 (8.1) years; body mass index, 25.0 (3.2) kg/m] were recruited. Each participant consumed the same breakfast at a slow (30 minutes) and fast (10 minutes) speed, on 2 separate days, in a randomized crossover design. Blood samples were collected in the fasting state and 3 hours postprandially during each eating condition. Appetite was assessed over the same period using visual analog scales. Blood concentrations of orexigenic hormone, ghrelin, and anorexigenic hormones, glucagon-like peptide-1 (GLP-1) and peptide YY (PYY), were determined. Daily food intake was measured, by food recall, after the slow and fast breakfast. Mixed-model repeated-measures analysis showed no eating condition or eating condition by time interaction effect on ghrelin, GLP-1, PYY, hunger, or fullness. Significant eating speed by time interaction effect on desire to eat was found (P=0.007). Desire to eat was lower at 60 minutes (P=0.007) after breakfast began during the slow versus fast eating condition. Eating speed at breakfast did not affect daily energy and macronutrient intake. Eating speed at breakfast did not affect postprandial ghrelin, GLP-1, PYY, hunger, and fullness values or daily energy and macronutrient intake. Desire to eat was lower at 60 minutes in the slow versus fast eating condition, but this result could not be explained by the changes in meal-related hormones measured in the study.