Expression and functional roles of Bradyrhizobium japonicum genes involved in the utilization of inorganic and organic sulfur compounds in free-living and symbiotic conditions.
Strains of Bradyrhizobium spp. form nitrogen-fixing symbioses with many legumes, including soybean. Although inorganic sulfur is preferred by bacteria in laboratory conditions, sulfur in agricultural soil is mainly present as sulfonates and sulfur esters. Here, we show that Bradyrhizobium japonicum and B. elkanii strains were able to utilize sulfate, cysteine, sulfonates, and sulfur-ester compounds as sole sulfur sources for growth. Expression and functional analysis revealed that two sets of gene clusters (bll6449 to bll6455 or bll7007 to bll7011) are important for utilization of sulfonates sulfur source. The bll6451 or bll7010 genes are also expressed in the symbiotic nodules. However, B. japonicum mutants defective in either of the sulfonate utilization operons were not affected for symbiosis with soybean, indicating the functional redundancy or availability of other sulfur sources in planta. In accordance, B. japonicum bacteroids possessed significant sulfatase activity. These results indicate that strains of Bradyrhizobium spp. likely use organosulfur compounds for growth and survival in soils, as well as for legume nodulation and nitrogen fixation.


The effects of age and social interactions on innate immunity in a leaf-cutting ant.
Both developmental and environmental factors shape investment in costly immune defences. Social insect workers have different selection pressures on their innate immune system compared to non-social insects because workers do not reproduce and their longevity affects the fitness of relatives. Furthermore, hygienic behavioural defences found in social insects can result in increased survival after fungal infection, although it is not known if there is modulation in physiological immune defence associated with group living vs. solitary living. Here we investigated whether physiological immune defence is affected by both age and the short-term presence or absence of nestmates in the leaf-cutting ant Acromyrmex octospinosus. We predicted that older ants would show immune senescence and that group living may result in prophylactic differences in immune defence compared to solitarily kept ants. We kept old and young workers alone or in nestmate groups for 48h and assayed a key innate immune system enzyme, expressing phenoloxidase (PO) and its stored precursor (proPO), a defence that acts immediately, i.e. it is constitutive. Short-term solitary living did not affect PO or proPO levels relative to group living controls and we found no evidence for immunosenescence in proPO. However, we found a significant increase in active PO in older workers, which is consistent with two non-mutually exclusive explanations: it could be an adaptive response or indicative of immunosenescence. Our results suggest that future studies of immunosenescence should consider both active immune effectors in the body, such as PO, and the stored potential to express immune defences, such as proPO.


N-Cadherin Is Critical for the Survival of Germ Cells, the Formation of Steroidogenic Cells, and the Architecture of Developing Mouse Gonads.
Normal gonad development assures the fertility of the individual. The properly functioning gonads must contain a sufficient number of the viable germ cells, possess a correct architecture and tissue structure, and assure the proper hormonal regulation. This is achieved by the interplay between the germ cells and different types of somatic cells. N-cadherin coded by the Cdh2 gene plays a critical role in this interplay. To gain an insight into the role of N-cadherin in the development of mouse gonads, we used the Cre-loxP system to knock out N-cadherin separately in two cell lines: the SF1+ somatic cells and the OCT4+ germ cells. We observed that N-cadherin plays a key role in the survival of both female and male germ cells. However, the N-cadherin is not necessary for the differentiation of the Sertoli cells or the initiation of the formation of testis cords or ovigerous cords. In the later stages of gonad development, N-cadherin is important for the maintenance of testis cord structure and is required for the formation of steroidogenic cells. In the ovaries, N-cadherin is necessary for the formation of the ovarian follicles. These results indicate that N-cadherin plays a major role in gonad differentiation, structuralization, and function.


Identifying Causal Risk Factors for Violence among Discharged Patients.
Structured Professional Judgement (SPJ) is routinely administered in mental health and criminal justice settings but cannot identify violence risk above moderate accuracy. There is no current evidence that violence can be prevented using SPJ. This may be explained by routine application of predictive instead of causal statistical models when standardising SPJ instruments. We carried out a prospective cohort study of 409 male and female patients discharged from medium secure services in England and Wales to the community. Measures were taken at baseline (pre-discharge), 6 and 12 months post-discharge using the Historical, Clinical and Risk-20 items version 3 (HCR-20v3) and Structural Assessment of Protective Factors (SAPROF). Information on violence was obtained via the McArthur community violence instrument and the Police National Computer. In a lagged model, HCR-20v3 and SAPROF items were poor predictors of violence. Eight items of the HCR-20v3 and 4 SAPROF items did not predict violent behaviour better than chance. In re-analyses considering temporal proximity of risk/ protective factors (exposure) on violence (outcome), risk was elevated due to violent ideation (OR 6.98, 95% CI 13.85-12.65, P<0.001), instability (OR 5.41, 95% CI 3.44-8.50, P<0.001), and poor coping/ stress (OR 8.35, 95% CI 4.21-16.57, P<0.001). All 3 risk factors were explanatory variables which drove the association with violent outcome. Self-control (OR 0.13, 95% CI 0.08-0.24, P<0.001) conveyed protective effects and explained the association of other protective factors with violence. Using two standardised SPJ instruments, predictive (lagged) methods could not identify risk and protective factors which must be targeted in interventions for discharged patients with severe mental illness. Predictive methods should be abandoned if the aim is to progress from risk assessment to effective risk management and replaced by methods which identify factors causally associated with violence.


Toward Developing a New Occupational Exposure Metric Approach for Characterization of Diesel Aerosols.
The extensive use of diesel-powered equipment in mines makes the exposure to diesel aerosols a serious occupational issue. The exposure metric currently used in U.S. underground noncoal mines is based on the measurement of total carbon (TC) and elemental carbon (EC) mass concentration in the air. Recent toxicological evidence suggests that the measurement of mass concentration is not sufficient to correlate ultrafine aerosol exposure with health effects. This urges the evaluation of alternative measurements. In this study, the current exposure metric and two additional metrics, the surface area and the total number concentration, were evaluated by conducting simultaneous measurements of diesel ultrafine aerosols in a laboratory setting. The results showed that the surface area and total number concentration of the particles per unit of mass varied substantially with the engine operating condition. The specific surface area (SSA) and specific number concentration (SNC) normalized with TC varied two and five times, respectively. This implies that miners, whose exposure is measured only as TC, might be exposed to an unknown variable number concentration of diesel particles and commensurate particle surface area. Taken separately, mass, surface area, and number concentration did not completely characterize the aerosols. A comprehensive assessment of diesel aerosol exposure should include all of these elements, but the use of laboratory instruments in underground mines is generally impracticable. The article proposes a new approach to solve this problem. Using SSA and SNC calculated from field-type measurements, the evaluation of additional physical properties can be obtained by using the proposed approach.


Chitosan-decorated selenium nanoparticles as protein carriers to improve the in vivo half-life of the peptide therapeutic BAY 55-9837 for type 2 diabetes mellitus.
As a potential protein therapeutic for type 2 diabetes mellitus (T2DM), BAY 55-9837 is limited by poor stability and a very short half-life in vivo. The purpose of this study was to construct a novel nanostructured biomaterial by conjugating BAY 55-9837 to chitosan-decorated selenium nanoparticles (CS-SeNPs) to prolong the in vivo half-life of BAY 55-9837 by reducing its renal clearance rate. BAY 55-9837-loaded CS-SeNPs (BAY-CS-SeNPs) were prepared, and their surface morphology, particle size, zeta potential, and structure were characterized. The stability, protein-loading rate, and in vitro release of BAY 55-9837 from CS-SeNPs were also quantified. Additionally, a sensitive high-performance liquid chromatography (HPLC) assay was developed for the quantification of BAY 55-9837 in mouse plasma. Thereafter, mice were injected via the tail vein with either BAY 55-9837 or BAY-CS-SeNPs, and the plasma concentration of BAY 55-9837 was determined via our validated HPLC method at different time intervals postinjection. Relevant in vivo pharmacokinetic parameters (half-life, area under the curve from time 0 to last sampling point, observed clearance) were then calculated and analyzed. BAY-CS-SeNPs were successfully synthesized, with diameters of approximately 200 nm. BAY-CS-SeNPs displayed good stability with a high protein-loading rate, and the release process of BAY 55-9837 from the CS-SeNPs lasted for over 70 hours, with the cumulative release reaching 78.9%. Moreover, the conjugation of CS-SeNPs to BAY 55-9837 significantly reduced its renal clearance to a rate of 1.56 mL/h and extended its half-life to 20.81 hours. In summary, our work provides a simple method for reducing the renal clearance rate and extending the half-life of BAY 55-9837 in vivo by utilizing CS-SeNPs as nanocarriers.


Salinibacter sensory rhodopsin: sensory rhodopsin I-like protein from a eubacterium.
Halobacterium salinarum sensory rhodopsin I (HsSRI), a dual receptor regulating both negative and positive phototaxis in haloarchaea, transmits light signals through changes in protein-protein interactions with its transducer, halobacterial transducer protein I (HtrI). Haloarchaea also have another sensor pigment, sensory rhodopsin II (SRII), which functions as a receptor regulating negative phototaxis. Compared with HsSRI, the signal relay mechanism of SRII is well characterized because SRII from Natronomonus pharaonis (NpSRII) is much more stable than HsSRI and HsSRII, especially in dilute salt solutions and is much more resistant to detergents. Two genes encoding SRI homologs were identified from the genome sequence of the eubacterium Salinibacter ruber. Those sequences are distantly related to HsSRI ( approximately 40% identity) and contain most of the amino acid residues identified as necessary for its function. To determine whether those genes encode functional protein(s), we cloned and expressed them in Escherichia coli. One of them (SrSRI) was expressed well as a recombinant protein having all-trans retinal as a chromophore. UV-Vis, low-temperature UV-Vis, pH-titration, and flash photolysis experiments revealed that the photochemical properties of SrSRI are similar to those of HsSRI. In addition to the expression system, the high stability of SrSRI makes it possible to prepare large amounts of protein and enables studies of mutant proteins that will allow new approaches to investigate the photosignaling process of SRI-HtrI.


The membrane-integrated transcriptional activator CadC of Escherichia coli senses lysine indirectly via the interaction with the lysine permease LysP.
In an acidic (pH 5.8) and lysine-rich environment Escherichia coli induces expression of the cadBA operon which encodes CadA, catalysing the decarboxylation of lysine to cadaverine, and CadB, the lysine/cadaverine antiporter. cadBA expression is dependent on CadC, a membrane-integrated transcriptional activator which belongs to the ToxR-like protein family and directly binds to the DNA in the cadBA promoter region. Here we describe that CadC senses the extracellular lysine not directly but indirectly requiring the interplay with the lysine permease LysP. Biochemical analyses of isolated CadC or the periplasmic domain of CadC (CadC188-512) revealed an unexpectedly low affinity for lysine, making it unlikely that CadC is a direct sensor for this substrate. Moreover, CadC hybrid proteins, in which the transmembrane domain or single amino acids were replaced, supported lysine-independent cadBA expression but retained a pH-dependent regulation. These CadC mutants were resistant to the effect of an overproduction of LysP, which represses cadBA expression in wild-type cells. Our results suggest a model according to which CadC is inactivated by an interaction with LysP at a low external lysine concentration. When lysine is abundantly available, the interaction between LysP and CadC is released, and CadC becomes susceptible to activation by low pH.


[Primary gastric lymphoma].
Primary Gastric Lymphoma is an uncommon malignancy among gastric malignancies. Histology of the Primary Gastric Lymphoma is varied and the extranodal marginal zone B-cells lymphoma is specially significant on account of its potential remission with antibiotic therapy. Observe the clinical characteristics of patients with Primary Gastric Lymphoma, assess the most relevant endoscopic findings, identify the factors that influence survival and evaluate the effects of therapy. The study is an observational, analytical, cross evaluation including 169 patients with histological diagnosis of Gastric Lymphoma, treated at the National Institute for Neoplastic Diseases, Lima, Peru, from January 1995 to December 2000. Staging was based on the Ann Arbor system, modified by Musshoff and histology, on the REAL-WHO classification. The statistical analysis included the student-t and the chi-square tests. Survival data were entered using the Kaplan Meier curves and prognosis factors, using the Cox regression test. The sample represents patients from the Peruvian Coast, with a mean age of 55 years old and slight predominance of female patients (54.4%). Signs and symptoms are unspecific. Clinical stage I-II corresponds to 75% of the patients. The endoscopic pattern of multiple ulcerated lesions is characteristic of the Gastric Lymphoma. A total of 71% of the patients with extranodal marginal zone B-cells lymphoma showed total remission of the disease with antibiotic therapy (5/7). The histological type of the Gastric Lymphoma in the 169 patients was as follows: Large, diffuse, B-cells Lymphoma, 137 patients, extranodal marginal zone B-cells lymphoma, 16 patients, peripheral T-cell Lymphoma, 6 patients, anaplastic large T-cell Lymphoma, 3 patients, undetermined Lymphoma, 3 patients, mantle cell Lymphoma, 2 patients, adult T-cell Lymphoma, 1 patient and follicular Lymphoma, 1 patient. Global survival after 36 months was of 61.34%, survival according to the histological type was of 92.31% for extranodal marginal zone B-cells Lymphomas, 62.21% for large, diffuse B-cells Lymphomas and 29.63% for T-cell Lymphomas. Survival after 36 months in patients in clinical stage I-II treated with chemotherapy, was of 82.16%, with surgery, 71.89% and with surgery and chemotherapy, 70.39, with similar results in all three groups (p: 0.6530). The groups classified according to the international index, showed a clear difference between them (p:0.0000). The univariate analysis revealed that Zubrod (p:0.0000) DHL (p:0.0073) disease remission (p:0.0000) stage (p:0.0000) treatment (p:0.0000) and location (p:0.0000) had statistical significance. Multivariate analysis showed that in the Cox regression model, remission (OR:13,342, p:0.0000) and location (OR:2.375, p:0.041) fall within the equation of such function. The multiple ulcerated lesions are characteristic of the Gastric Lymphoma. Remission of the disease in the extranodal marginal zone B-cells Lymphoma is evidenced with the use of antibiotic therapy (5/7). Chemotherapy in patients with EC I-II achieves survival results similar to those treated with surgery and with a combination of both. Validity of the international index is confirmed and the multivariate analysis proved that remission and location of the disease have statistical significance.


TREC to WHERE? Transdisciplinary Research on Energetics and Cancer.
When information is exchanged across disciplinary boundaries, resources are shared, and discipline-specific approaches are altered to achieve a common scientific goal, we create a new intellectual space for transdisciplinary research. This approach, fostered heavily by multiple NCI-funded initiatives, has the potential to forge new understanding of major public health issues. By breaking down disciplinary barriers, we work toward making real, meaningful, and lasting forward motion in addressing key public health issues. One of the transdisciplinary initiatives of the NCI is TREC: Transdisciplinary Research on Energetics and Cancer. In this article, we review the goals and scope of TREC, as well as the ways in which the initiative promotes transdisciplinary science. A particular focus is on multiple examples of the most unique aspect of the initiative: the funding of developmental projects across multiple TREC centers, toward the goal of incubating high-risk science that has the potential to translate into major leaps forward in understanding energetics in cancer. As we enter an era of greater focus on investigator-initiated science, new approaches may be needed to ensure that the peer review process is not solely organized along disciplinary lines. Inclusion of expertise regarding transdisciplinarity, as well as representation from multiple scientific disciplines within a panel, may allow transdisciplinary research to receive an educated hearing. The body of researchers trained to work in a transdisciplinary research space is ideally suited to address these challenges.


Current perspectives on the role of biomechanical loading and genetics in development of disc degeneration and low back pain; a narrative review.
Degenerative changes in the disc have long been of interest; they are thought to be strongly associated with low back pain and caused by inappropriate loading or through injury. However, independent of the magnitude of occupational spinal loading, twin studies find that the heritability of lumbar disc degeneration is 34-74%. This finding has led to intensive searches for susceptibility genes; some genes associated with disc degeneration have been identified, though all with small effects on the degenerative process. The complex nature of degenerative changes suggests that many different genes are involved, and that interactions with environmental factors are influential in progression of degeneration. Low back pain itself also appears heritable (30-46%). The most important clinical question though, is not how discs degenerate but is disc degeneration related to low back pain. Imaging studies find many people with degenerate discs or even with discs showing pathological features such as herniations, are asymptomatic. However results are obscured by the lack of consistent definitions of the phenotypes of disc degeneration and of low back pain. Epidemiological studies could help disentangle these complex relationships, but they will only be successful once consistent classifications and phenotypes of both disc degeneration and low back pain are developed.


Guilt in the Treatment of Posttraumatic Stress Disorder Among Active Duty Military Personnel.
The current study examined the role of trauma-related guilt on posttraumatic stress disorder (PTSD) symptom change during prolonged exposure therapy (PE) as well as the efficacy of PE in reducing three dimensions of guilt (responsibility, wrongdoing, and lack of justification) during treatment. Participants were 331 active duty U.S. military personnel seeking treatment for PTSD who were randomized to one of four groups: massed PE (10 sessions delivered over 2 weeks), spaced PE (10 sessions delivered over 8 weeks), present-centered therapy (PCT; 10 sessions delivered over 8 weeks), or minimal contact control (MCC; weekly therapist phone check-in for 4 weeks). The results showed that baseline guilt did not predict reductions in PTSD symptoms for spaced PE or for PCT, ps = .178-.387, ds = -0.02-0.07. Treatment condition (massed PE vs. MCC; spaced PE vs. PCT) did not moderate reductions in guilt for spaced PE versus PCT. Guilt decreased significantly over treatment in all groups, p < .001 to p = .038, ds = -0.19 to -0.42, except concerning justification in the spaced PE and PCT groups, p = .140, d = -0.10. The findings suggest that guilt may be reduced significantly following active PTSD treatment and attention control and that PTSD recovery is not impacted by baseline levels of trauma-related guilt in military personnel with PTSD, although reported levels of guilt were low to moderate in this sample.


Genetic basis and evolution of rapid cycling in railway populations of tetraploid Arabidopsis arenosa.
Spatially structured plant populations with diverse adaptations provide powerful models to investigate evolution. Human-generated ruderal habitats are abundant and low-competition, but are challenging for plants not adapted to them. Ruderal habitats also sometimes form networked corridors (e.g. roadsides and railways) that allow rapid long-distance spread of successfully adapted variants. Here we use transcriptomic and genomic analyses, coupled with genetic mapping and transgenic follow-up, to understand the evolution of rapid cycling during adaptation to railway sites in autotetraploid Arabidopsis arenosa. We focus mostly on a hybrid population that is likely a secondary colonist of a railway site. These mountain railway plants are phenotypically similar to their cosmopolitan cousins. We thus hypothesized that colonization primarily involved the flow of adaptive alleles from the cosmopolitan railway variant. But our data shows that it is not that simple: while there is evidence of selection having acted on introgressed alleles, selection also acted on rare standing variation, and new mutations may also contribute. Among the genes we show have allelic divergence with functional relevance to flowering time are known regulators of flowering, including FLC and CONSTANS. Prior implications of these genes in weediness and rapid cycling supports the idea that these are "evolutionary hotspots" for these traits. We also find that one of two alleles of CONSTANS under selection in the secondary colonist was selected from rare standing variation in mountain populations, while the other was introgressed from the cosmopolitan railway populations. The latter allele likely arose in diploid populations over 700km away, highlighting how ruderal populations could act as allele conduits and thus influence local adaptation.


Chronic Diffuse Sclerosing Osteomyelitis of the Mandible.
Chronic diffuse sclerosing osteomyelitis (CDSO) is a disease that presents a rather unusual clinicoradiographic picture creating diagnostic and therapeutic problems. Involvement of the temporomandibular joint is not frequently observed in CDSO. We report a unique case of a 37-year-old male with cyclic episodes of insidious swelling and progressive trismus for the past 4 years. Each episode was followed by spontaneous remission after 2 weeks. Radiographic examination detected unilateral coarse trabeculae with ground-glass appearance of the left hemimandible and resorption of ipsilateral condyle. Computed tomography demonstrated endosteal sclerosis, cortical thickening, and condylar resorption, while magnetic resonance imaging revealed altered marrow intensity with postcontrast enhancement of the surrounding musculature. Bone histopathology was consistent with the diagnosis of osteomyelitis. Palliative antibiotic therapy and regular follow-up were recommended. Of particular interest in the present case is ipsilateral condylar resorption not associated with any dermato-skeletal conditions, which is uncommon in CDSO of the mandible.


Formal intergenerational mentoring at Australian Men's Sheds: a targeted survey about mentees, mentors, programmes and quality.
Intergenerational mentoring enables a purposeful exchange of skills and knowledge to enhance individual and social outcomes for sub-groups at risk of health and social disparities. Male intergenerational mentoring may be an approach to help address these disparities in young men. Over 1000 Men's Sheds operate in Australia with 39% providing some form of mentoring mainly to youth. Yet, little is known about the variables intrinsic to creating and running quality programmes. This study aimed to identify the characteristics of formal intergenerational mentoring programmes, review their quality against the Australian Youth Mentoring Network (AYMN) quality benchmarks, and identify the factors that predict quality in these programmes. All known Australian Men's Sheds were invited to participate in an online cross-sectional survey. Forty sheds with formal mentor programmes completed the survey for a total of 387 mentees (mean = 9.7 mentees/programme), the majority being male. The majority of mentor programme facilitators were unpaid male volunteers aged 61 years and older, and programmes were unfunded. Promoting social and emotional well-being of the mentees was the primary focus in more than half of the programmes, and working on a shared construction project was the most common activity. Respondents rated the three most important factors that influenced programme effectiveness as being: (i) meaningful activities; (ii) mentors' approach; and (iii) a safe environment. Univariate analyses revealed that mentoring programmes that had a system in place for screening mentors, trained mentors and evaluated the programme were most likely to rate highly against the AYMN quality benchmarks.


Epigenetic reprogramming in cancer.
The demonstration of induced pluripotency and direct lineage conversion has led to remarkable insights regarding the roles of transcription factors and chromatin regulators in mediating cell state transitions. Beyond its considerable implications for regenerative medicine, this body of work is highly relevant to multiple stages of oncogenesis, from the initial cellular transformation to the hierarchical organization of established malignancies. Here, we review conceptual parallels between the respective biological phenomena, highlighting important interrelationships among transcription factors, chromatin regulators, and preexisting epigenetic states. The shared mechanisms provide insights into oncogenic transformation, tumor heterogeneity, and cancer stem cell models.


Flanking residues help determine whether a hydrophobic segment adopts a monotopic or bitopic topology in the endoplasmic reticulum membrane.
Proteins interacting with membranes via a single hydrophobic segment can be classified as either monotopic or bitopic. Here, we probe the topology of a membrane-attached enzyme, the ε isoform of human diacylglycerol kinase (DGKε), when inserted into rough microsomes and compare it with the monotopic membrane protein mouse caveolin-1. In contrast to previous findings, the N-terminal hydrophobic stretch in DGKε attains a bitopic rather than a monotopic topology in our experimental system. In addition, we find that charged flanking residues as well as proline residues embedded in the hydrophobic segment are important determinants of monotopic versus bitopic topology.


Lamb Performance on Island Pastures in Northern Norway.
The Norwegian sheep industry is based on utilization of "free" rangeland pasture resources. Use of mountain pastures is dominating, with about two million sheep grazing these pastures during summer. Regional challenges related to e.g., loss of sheep to large carnivores make farmers think differently. The Norwegian coastline is among the longest globally and is scattered with islets and islands. Alone along the coast of Nordland county, it is estimated more than 14,000 islands. Use of islands for summer pasture is an alternative but there is a limited knowledge about such a management system. In this study, we examined lambs' average daily gain on island pastures at the coast of Norway. In total 230 lambs on three islands (Sandvær, Sjonøya, and Buøya), with varying pasture quality and stocking rate, for 3 years (2012, 2013, and 2014). At Sandvær as much as 92% of the island was characterized as high nutritional value while at Sjonøya and Buøya only 15%, was characterized high nutritional value. We found an average daily lamb growth rate of 0.320 kg d-1. Lambs on Sandvær had a higher daily gain (P < 0.05) than those on Sjonøya and Buøya, and lambs' average daily gain was significantly lower (P < 0.05) in 2013 compared to 2012 and 2014. We conclude that with a dynamic and adaptive management strategy there is a potential to utilize islands for sheep grazing during summer.


Hypothalamic S1P/S1PR1 axis controls energy homeostasis in Middle-Aged Rodents: the reversal effects of physical exercise.
Recently, we demonstrated that the hypothalamic S1PR1/STAT3 axis plays a critical role in the control of food consumption and energy expenditure in rodents. Here, we found that reduction of hypothalamic S1PR1 expression occurs in an age-dependent manner, and was associated with defective thermogenic signaling and weight gain. To address the physiological relevance of these findings, we investigated the effects of chronic and acute exercise on the hypothalamic S1PR1/STAT3 axis. Chronic exercise increased S1PR1 expression and STAT3 phosphorylation in the hypothalamus, restoring the anorexigenic and thermogenic signals in middle-aged mice. Acutely, exercise increased sphingosine-1-phosphate (S1P) levels in the cerebrospinal fluid (CSF) of young rats, whereas the administration of CSF from exercised young rats into the hypothalamus of middle-aged rats at rest was sufficient to reduce the food intake. Finally, the intracerebroventricular (ICV) administration of S1PR1 activators, including the bioactive lipid molecule S1P, and pharmacological S1PR1 activator, SEW2871, induced a potent STAT3 phosphorylation and anorexigenic response in middle-aged rats. Overall, these results suggest that hypothalamic S1PR1 is important for the maintenance of energy balance and provide new insights into the mechanism by which exercise controls the anorexigenic and thermogenic signals in the central nervous system during the aging process.


Activation of p53 by nutlin-3a induces apoptosis and cellular senescence in human glioblastoma multiforme.
Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor in adults. Despite concerted efforts to improve current therapies and develop novel clinical approaches, patient survival remains poor. As such, increasing attention has focused on developing new therapeutic strategies that specifically target the apoptotic pathway in order to improve treatment responses. Recently, nutlins, small-molecule antagonists of MDM2, have been developed to inhibit p53-MDM2 interaction and activate p53 signaling in cancer cells. Glioma cell lines and primary cultured glioblastoma cells were treated with nutlin-3a. Nutlin-3a induced p53-dependent G1- and G2-M cell cycle arrest and apoptosis in glioma cell lines with normal TP53 status. In addition, nutlin-arrested glioma cells show morphological features of senescence and persistent induction of p21 protein. Furthermore, senescence induced by nutlin-3a might be depending on mTOR pathway activity. In wild-type TP53 primary cultured cells, exposure to nutlin-3a resulted in variable degrees of apoptosis as well as cellular features of senescence. Nutlin-3a-induced apoptosis and senescence were firmly dependent on the presence of functional p53, as revealed by the fact that glioblastoma cells with knockdown p53 with specific siRNA, or cells with mutated or functionally impaired p53 pathway, were completely insensitive to the drug. Finally, we also found that nutlin-3a increased response of glioma cells to radiation therapy. The results provide a basis for the rational use of MDM2 antagonists as a novel treatment option for glioblastoma patients.


Plasmodium chabaudi chabaudi malaria parasites can develop stable resistance to atovaquone with a mutation in the cytochrome b gene.
Plasmodium falciparum, has developed resistance to many of the drugs in use. The recommended treatment policy is now to use drug combinations. The atovaquone-proguanil (AP) drug combination, is one of the treatment and prophylaxis options. Atovaquone (ATQ) exerts its action by inhibiting plasmodial mitochondria electron transport at the level of the cytochrome bc1 complex. Plasmodium falciparum in vitro resistance to ATQ has been associated with specific point mutations in the region spanning codons 271-284 of the cytochrome b gene. ATQ -resistant Plasmodium yoelii and Plasmodium berghei lines have been obtained and resistant lines have amino acid mutations in their CYT b protein sequences. Plasmodium chabaudi model for studying drug-responses and drug-resistance selection is a very useful rodent malaria model but no ATQ resistant parasites have been reported so far. The aim of this study was to determine the ATQ sensitivity of the P. chabaudi clones, to select a resistant parasite line and to perform genotypic characterization of the cytb gene of these clones. To select for ATQ resistance, Plasmodium. chabaudi chabaudi clones were exposed to gradually increasing concentrations of ATQ during several consecutive passages in mice. Plasmodium chabaudi cytb gene was amplified and sequenced. ATQ resistance was selected from the clone AS-3CQ. In order to confirm whether an heritable genetic mutation underlies the response of AS-ATQ to ATQ, the stability of the drug resistance phenotype in this clone was evaluated by measuring drug responses after (i) multiple blood passages in the absence of the drug, (ii) freeze/thawing of parasites in liquid nitrogen and (iii) transmission through a mosquito host, Anopheles stephensi. ATQ resistance phenotype of the drug-selected parasite clone kept unaltered. Therefore, ATQ resistance in clone AS-ATQ is genetically encoded. The Minimum Curative Dose of AS-ATQ showed a six-fold increase in MCD to ATQ relative to AS-3CQ. A mutation was found on the P. chabaudi cytb gene from the AS-ATQ sample a substitution at the residue Tyr268 for an Asn, this mutation is homologous to the one found in P. falciparum isolates resistant to ATQ.


Quantitative evaluation of macrophage aggregates in brook trout Salvelinus fontinalis and rainbow trout Oncorhynchus mykiss.
Macrophage aggregates (MAs) occur in various organs of fishes, especially the kidney, liver and spleen, and contain melanin, ceroid/lipofuscin and hemosiderin pigments. They have been used as indicators of a number of natural and anthropogenic stressors. Macrophage aggregates occur in salmonids but are poorly organized, irregularly shaped, and are generally smaller than those in derived teleosts. These features complicate quantification, and thus these fishes have seldom been used in studies correlating MAs with environmental stressors. To alleviate these complications, we developed color filtering algorithms for use with the software package ImagePro Plus (Media Cybernetics) that select and quantify pigmented area (i.e. colors ranging from gold to brown to black) in tissue sections. Image analysis results compared well with subjective scoring when tested on brook trout Salvelinus fontinalis and rainbow trout Oncorhynchus mykiss captured from high-elevation lakes or hatcheries. Macrophage aggregate pigments correlated positively with age and negatively with condition factor. Within individual fish, pigmentation correlated positively among organs, suggesting that the kidney, liver or spleen are suitable indicator organs. In age-matched fishes, MA pigments were not different between hatcheries and lakes in the organs examined. Between lakes, differences in pigments were observed in the kidney and spleen, but were not explained by age, condition factor, sex or maturation state. Our results indicate that quantification of the area occupied by MA pigments is an efficient and accurate means of evaluating MAs in salmonid organs and that organ pigmentation correlates with age and condition factor, as seen in studies with more derived fishes.


Data-based modelling and proportional-integral-plus (pip) control of nitrate in an activated sludge benchmark.
This paper presents the result of an investigation into the Proportional Integral Plus (PIP) control of nitrate in the second zone of an activated sludge benchmark. A data-based reduced order model is used as the control model and identified using the Simplified Refined Instrumental Variable (SRIV) identification and estimation algorithm. The PIP control design is based on the Non Minimum State Space (NMSS) form and State Variable Feedback (SVF) methodology. The PIP controller is tested against dynamic load disturbances and compared with the response of a well tuned PI controller.


Electronic medical record use by office-based physicians and their practices: United States, 2007.
This report presents information on the adoption of electronic medical records (EMRs) by office-based physicians in 2007. Percentages of medical practices and physicians within practices using EMR systems are presented by selected physician and practice characteristics. Data from the physician induction interviews of the 2007 National Ambulatory Medical Care Survey (NAMCS) are presented. NAMCS is based upon a national probability sample of nonfederal office-based physicians who saw patients in an office setting. Sample data were weighted to produce national estimates of office-based physician characteristics and their practices. In 2007, 34.8 percent of office-based physicians reported using any EMR (all electronic or partially electronic medical record) system, which represented a 19.2 percent increase since 2006 and a 91.2 percent increase since 2001. Starting in 2005, NAMCS included additional questions about features of electronic record systems making it possible to categorize systems as basic or fully functional using similar definitions developed by health information technology (HIT) experts. Fully functional systems are a subset of basic systems. Some systems do not meet the requirements. In 2007, 11.8 percent of physicians had systems meeting the criteria of basic systems (95% CI: 9.6-13.9), unchanged from 2006 (10.5 percent). The percentage of office-based physicians with systems meeting the definition of fully functional (3.8 percent, 95% CI: 2.6-5.0) was similar to the 2006 percentage (3.1 percent). Physicians in practices with 11 or more physicians were most likely to use any EMR system (74.3 percent), whereas physicians in solo practice were least likely to use EMRs (20.6 percent). EMR use was higher among physicians in multi-specialty practices (52.5 percent) than in solo or single-specialty practices (30.3 percent). EMR use was inversely associated with physician age. If those without EMR systems in 2007 with definite plans to install one actually do so, 53.6 percent of physicians will have some type of an EMR system in 2010.


Social dimensions of healing: a longitudinal study of outcomes of medical management of headaches.
The social sciences have made few direct empirical contributions to the understanding of 'non-specific' benefits of treatment and generally the symbolic healing of indigenous non-Western medicine has received most attention in this field. This paper reports some results of a wider study of neurological clinics in England in which it is shown that a sample attending for headaches experienced considerable improvement in symptoms when followed up 1 year after attendance. Most of this improvement appeared not to be due to any intended treatments received at the clinics but could be attributed to the quality of patients' immediate responses to clinic attendance as assessed from reach interviews conducted after their consultations. This relationship between immediate 'satisfied' response and subsequent symptomatic improvement is interpreted in terms of the general levels of expectancy and sense of potential control achieved by obtaining referral to a specialist which directly enhanced recovery in those patients who felt the doctor's actions to be directly relevant to their personal concerns. Disappointment with the doctor reduced the 'non-specific' therapeutic benefits of the hospital referral. The intimate connections of patient satisfaction, treatment received and subsequent outcomes need more careful consideration in social studies of Western medicine.


Discontinuation of yellow jacket venom immunotherapy: follow-up of 75 patients by means of deliberate sting challenge.
Venom immunotherapy is effective in preventing systemic reactions in patients with a history of an anaphylactic reaction to Hymenoptera stings. It is uncertain how long venom immunotherapy should be continued. We evaluated whether the duration of venom immunotherapy given to yellow jacket-sensitive patients related to the risk of an anaphylactic reaction to a later sting. Seventy-five yellow jacket-sensitive patients (29 male and 46 female) received a median number of three in-hospital sting challenges from a live insect in 3 subsequent years after discontinuation of venom immunotherapy. An anaphylactic reaction to one or more of the sting challenges was considered a relapse. We analyzed whether patients with and patients without a relapse differed in terms of gender, age, preimmunotherapy skin test data, preimmunotherapy level of venom-specific IgE, severity of the field-sting reaction that preceded immunotherapy, severity of the reaction to the sting challenge that preceded immunotherapy, adverse reactions to immunotherapy, changes in IgE and IgG4 levels during immunotherapy, duration of immunotherapy, and presence of venom-specific IgE after cessation of therapy. Venom immunotherapy was given for a median duration of 40 months (range, 7 to 120 months). Relapses were observed in six patients. In two of them, a rather severe anaphylactic reaction was observed after the second sting challenge. No relation was found between duration of venom immunotherapy and relapse risk. The relapse rate was higher among patients with high levels of specific IgE before and after immunotherapy. During therapy, the mean level of specific IgE decreased. This decline persisted in the 3 following years. No relapses of sting reactions were observed among patients without detectable specific IgE. Discontinuation of venom immunotherapy appears safe for patients with pretreatment IgE antibodies if these antibodies can no longer be detected during immunotherapy. For the remaining patients, a treatment period of 3 years may suffice. After discontinuation of immunotherapy, a clinical sting challenge can be considered to estimate the patient's current grade of hypersensitivity.


SU-E-I-50: The ACR CT Dose Index Registry: Implementation Challenges and Preliminary Data.
To successfully register our institution into the American College of Ragiology (ACR) CT Dose Index Registry (DIR), to discuss the technical and implementation challenges of doing so, and to review preliminary results. TRIAD is the American College of Radiology Imaging Network's (ACRIN) image acquisition and management software, with the purpose of receiving and then transmitting DICOM structured dose report files from institutional CT scanners around the country. TRIAD was installed onto a virtual machine server at our institution so that anonymized and encrypted DICOM structured dose report files from our 6 CT scanners could be sent to the TRIAD Application Server at ACR. Doing so required collaboration between ACR and IT, PACS personnel and the physicist on site. Implementation involved several challenges, such as software installation and data transmission consistency problems. Since numerous institutions are involved, the ACR required an exam mapping process via the Radlex playbook to unify the protocol classification. These challenges have been overcome and data is being successfully transmitted to and analyzed by the ACR. The first report comparing dose data (CTDI and DLP by examination and by scan) between our site and others around the region and country was made available recently. For each exam, the report includes boxplots and histogram data for a variety of standard protocols. For example, for a CT head exam, our median CTDIvol and DLP were 44 mGy and 736 mGy-cm compared with 64 mGy and 844 mGy-cm respectively, for all other facilities registered in the DIR. The ACR CT DIR registry is a useful tool for dose data mining and will eventually establish national benchmarks for CT dose indices. Our experience will allow others to anticipate these challenges and have potential solutions available for when they do arise.


Xwnt-8 and lithium can act upon either dorsal mesodermal or neurectodermal cells to cause a loss of forebrain in Xenopus embryos.
When Xenopus gastrulae are made to misexpress Xwnt-8, or are exposed to lithium ions, they develop with a loss of anterior structures. In the current study, we have characterized the neural defects produced by either Xwnt-8 or lithium and have examined potential cellular mechanisms underlying this anterior truncation. We find that the primary defect in embryos exposed to lithium at successively earlier stages during gastrulation is a progressive rostral to caudal deletion of the forebrain, while hindbrain and spinal regions of the CNS remain intact. Misexpression of Xwnt-8 during gastrulation produces an identical loss of forebrain. Our results demonstrate that lithium and Wnts can act upon either prospective neural ectodermal cells, or upon dorsal mesodermal cells, to cause a loss of anterior pattern. Specifically, ectodermal cells isolated from lithium- or Wnt-exposed embryos are unable to form anterior neural tissue in response to inductive signals from normal dorsal mesoderm. In addition, although dorsal mesodermal cells from lithium- or Wnt-exposed embryos are specified properly, and produce normal levels of the anterior neural inducing molecules noggin and chordin, they show a greatly reduced capacity to induce anterior neural tissue in conjugated ectoderm. Taken together, our results are consistent with a model in which Wnt- or lithium-mediated signals can induce either mesodermal or ectodermal cells to produce a dominant posteriorizing morphogen which respecifies anterior neural tissue as posterior.


A rare occupation causing mesothelioma: mechanisms and differential etiology.
In a mesothelioma lawsuit, the Public Prosecutor commissioned an expert evidence on the legal accountability for the disease, because the patient experienced multiple exposures to asbestos in both occupational and environmental settings. To collect information on asbestos exposure from all available sources and to quantify the contribution of each source of exposure as a percentage of the total risk. We retrieved information on jobs done and asbestos exposure from a work colleague and a database maintained by the National Institute for Insurance of Occupational Accidents/Diseases, respectively. Information on environmental exposure was searched through the scientific literature. The contribution of each source of exposure was quantified with a method of risk apportionment, taking into account time elapsed since first and last exposure, intensity and frequency of exposure and carcinogenic potency of asbestos fiber mix. The subject worked in the maintenance of railway electrification system. The mechanical compression stress induced on the ballast during passage of trains released chrysotile (from fragmented stones) and crocidolite (through abrasive action of crushed gravel on the underbody of rolling stocks insulated with friable crocidolite). Despite the low cumulative exposure (about 2 ff×years/cc), 99% of the mesothelioma risk was attributable to the work done because of the high content of crocidolite of inhaled asbestos. The report of an uncommon source of occupational asbestos exposure and a scientifically based method to allocate mesothelioma risk among multiple exposure could help to recognize mesothelioma as occupational disease in the workers employed in maintenance of the railway electrification system under the Italian National Railways.


Crystal structure of the heteromolecular chaperone, AscE-AscG, from the type III secretion system in Aeromonas hydrophila.
The putative needle complex subunit AscF forms a ternary complex with the chaperones AscE and AscG in the type III secretion system of Aeromonas hydrophila so as to avoid premature assembly. Previously, we demonstrated that the C-terminal region of AscG (residues 62-116) in the hetero-molecular chaperone, AscE-AscG, is disordered and susceptible to limited protease digestion. Here, we report the crystal structure of the ordered AscG(1-61) region in complex with AscE at 2.4 Å resolution. Helices α2 and α3 of AscE in the AscE-AscG(1-61) complex assumes a helix-turn-helix conformation in an anti-parallel fashion similar to that in apo AscE. However, in the presence of AscG, an additional N-terminal helix α1 in AscE (residues 4-12) is observed. PscG or YscG in the crystal structures of PscE-PscF-PscG or YscE-YscF-YscG, respectively, assumes a typical tetratricopeptide repeat (TPR) fold with three TPR repeats and one C-terminal capping helix. By comparison, AscG in AscE-AscG(1-61) comprises three anti-parallel helices that resembles the N-terminal TPR repeats in the corresponding region of PscG or YscG in PscE-PscF-PscG or YscE-YscF-YscG. Thermal denaturation of AscE-AscG and AscE-AscG(1-61) complexes demonstrates that the C-terminal disordered region does not contribute to the thermal stability of the overall complex. The N-terminal region of the AscG in the AscE-AscG complex is ordered and assumes a structure similar to those in the corresponding regions of PscE-PscG-PscF or YscE-YscF-YscG complexes. While the C-terminal region of AscG in the AscE-AscG complex is disordered and will assume its structure only in the presence of the substrate AscF. We hypothesize that AscE act as a chaperone of the chaperone to keep AscG in a stable but partially disordered state for interaction with AscF.


MM2-cortical-type sporadic Creutzfeldt-Jakob disease with early stage cerebral cortical pathology presenting with a rapidly progressive clinical course.
We report the case of a 67-year-old man with MM2-cortical-type sporadic Creutzfeldt-Jakob disease (sCJD) with a rapidly progressive clinical course of 5 months. Initial symptoms were progressive memory disturbance and dementia. MRI revealed high signal-intensity lesions on diffusion-weighted images in the bilateral frontal and occipital cortices. Myoclonus and periodic sharp-wave complexes on the electroencephalogram were observed in the early disease stage. The clinical diagnosis was typical sCJD. Neuropathologic examination at autopsy showed widespread, characteristic cerebral neocortical involvement with large confluent vacuole-type spongiform change. Spongiform degeneration was also evident in the striatum and medial thalamus. In the cerebellar cortex, slight depletion of Purkinje neurons was evident without spongiform change in the molecular layer or apparent neuron loss in the granule cell layer. The inferior olivary nucleus showed slight hypertrophic astrocytosis without neuron loss. Prion protein (PrP) immunostaining showed widespread, characteristic perivacuolar-type PrP deposits with irregular plaque-like PrP deposits in the cerebral neocortex, striatum and medial thalamus. We believe this patient showed early-stage cerebral cortical pathology of MM2-cortical-type sCJD, which may provide clues regarding the pathologic progression of this rare sCJD subtype. Although MM2-cortical-type sCJD generally shows slow progression without myoclonus or periodic sharp-wave complexes, the present patient showed a rapidly progressive clinical course similar to that of MM1-type sCJD.


Clinical spectrum and prognostic factors of possible UIP pattern on high-resolution CT in patients who underwent surgical lung biopsy.
Few studies have reported the diagnostic variability in patients with a possible usual interstitial pneumonia (UIP) pattern on high-resolution CT (HRCT) who underwent surgical lung biopsy (SLB), and the prognostic factors for these patients have not been fully evaluated. We retrospectively investigated the frequency of idiopathic pulmonary fibrosis (IPF) and prognostic factors in patients with possible UIP pattern on HRCT. Consecutive patients who had a possible UIP pattern on HRCT, underwent SLB, and had a diagnosis of IIPs before SLB were retrospectively recruited from 10 hospitals. Diagnoses were made based on multidisciplinary discussion using the criteria for current IPF guidelines and multidisciplinary classification for IIPs in each hospital. 179 patients who underwent SLB were enrolled. The diagnoses were IPF in 91 patients (51%), unclassifiable IIPs in 47 (26%), idiopathic NSIP in 18 (10%), and chronic hypersensitivity pneumonia in 17 (9%). One-year FVC changes showed significant differences between IPF and non-IPF (-138.6 mL versus 18.2 mL, p = 0.014). Patients with IPF had a worse mortality than those with non-IPF (Logrank test, p = 0.025). Multivariable Cox regression analysis demonstrated that diagnoses of IPF (HR, 2.961; 95% CI, 1.183-7.410; p = 0.02), high modified MRC score (HR, 1.587; 95% CI, 1.003-2.510; p = 0.049), and low %FVC (HR, 0.972; 95% CI, 0.953-0.992; p = 0.005). About a half of patients with a possible UIP pattern on HRCT had diagnoses other than IPF, and patients with IPF had a worse mortality than those with an alternative diagnosis. We reaffirmed that multidisciplinary discussion is crucial in patients with possible UIP pattern on HRCT.


Organometallic single-molecule electronics: tuning electron transport through X(diphosphine)2FeC4Fe(diphosphine)2X building blocks by varying the Fe-X-Au anchoring scheme from coordinative to covalent.
A series of X(depe)2FeC≡C-C≡CFe(depe)2X complexes (depe =1,2-bis(diethylphosphino)ethane; X = I 1, NCMe 2, N2 3, C2H 4, C2SnMe3 5, C4SnMe3 6, NCSe 7, NCS 8, CN 9, SH 10, and NO2 11) was designed to study the influence of the anchor group on organometallic molecular transport junctions to achieve high-conductive molecular wires. The FeC4Fe core is electronically functional due to the redox-active Fe centers and sp-bridging ligands allowing a strong electronic delocalization. 1-11 were characterized by elemental analyses, X-ray diffraction, cyclic voltammetry, NMR, IR, and Raman spectroscopy. DFT calculations on model compounds gave the HOMO/LUMO energies. 5-9 were investigated in mechanically controllable break-junctions. For 9, unincisive features at 8.1 × 10(-7) G0 indicate that sterical reasons prevent stable junctions to form or that the coordinative binding motif prohibits electron injection. 7 and 8 with the hitherto unexploited coordinatively binding end groups NCSe and NCS yielded currents of 1.3 × 10(-9) A (7) and 1.8 × 10(-10) A (8) at ±1.0 V. The SnMe3 in 5 and 6 splits off, yielding junctions with covalent C-Au bonds and currents of 6.5 × 10(-7) A (Au-5'-Au) or 2.1 × 10(-7) A (Au-6'-Au). Despite of a length of almost 2 nm, the Au-5'-Au junction reaches 1% of the maximum current assuming one conductance channel in quantum point contacts. Additionally, the current noise in the transport data is considerably reduced for the covalent C-Au coupling compared to the coordinative anchoring of 7-9, endorsing C-Au coupled organometallic complexes as excellent candidates for low-ohmic molecular wires.


Interventions for preventing distal intestinal obstruction syndrome (DIOS) in cystic fibrosis.
Cystic fibrosis (CF) is the most common, life-limiting, genetically inherited disease. It affects multiple organs, particularly the respiratory system. However, gastrointestinal problems such as constipation and distal intestinal obstruction syndrome (DIOS) are also important and well-recognised complications in CF. They share similar symptoms e.g. bloating, abdominal pain, but are distinct conditions. Constipation occurs when there is gradual faecal impaction of the colon, but DIOS occurs when there is an accumulation of faeces and sticky mucus, forming a mass in the distal part of the small intestine. The mass may partially block the intestine (incomplete DIOS) or completely block the intestine (complete DIOS). Symptoms of DIOS can affect quality of life and other aspects of CF health, such as airway clearance, exercise, sleep and nutritional status. Treatment of constipation and prevention of complete bowel obstruction are required for gastrointestinal management in CF. However, many different strategies are used in clinical practice and there is a lack of consensus. The importance of this topic was highlighted in a recent research priority setting exercise by the James Lind Alliance. To evaluate the effectiveness and safety of laxative agents of differing types for preventing DIOS (complete and incomplete) in children and adults with CF. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register comprising references identified from comprehensive electronic database searches and handsearches of relevant journals and abstract books of conference proceedings. Date of search: 22 May 2018.We also searched online trial registries. Date of last search: 10 June 2018. Randomised and quasi-randomised controlled parallel trials comparing laxative therapy for preventing DIOS (including osmotic agents, stimulants, mucolytics and substances with more than one action) at any dose to placebo, no treatment or an alternative laxative therapy, in people of any age with pancreatic sufficient or insufficient CF and any stage of lung disease. Randomised cross-over trials were judged on an individual basis. Two authors independently assessed trials for inclusion, extracted outcome data and performed a risk of bias assessment for the included data. We judged the quality of the evidence using GRADE criteria. We included one cross-over trial (17 participants) with a duration of 12 months, in which participants were randomly allocated to either cisapride (a gastro-prokinetic agent) or placebo for six months each. The trial had an unclear risk of bias for most domains but had a high risk of reporting bias.Radiograph scores revealed no difference in occurrence of DIOS between cisapride and placebo (narrative report, no data provided). There were no adverse effects. Symptom scores were the only secondary outcome within the review that were reported. Total gastrointestinal symptom scores favoured cisapride with a statistically significant mean difference (MD) of -7.60 (95% confidence interval (CI) -14.73 to -0.47). There was no significant difference at six months between cisapride and placebo for abdominal distension, MD -0.90 (95% CI -2.39 to 0.59) or abdominal pain, MD -0.4 (95% CI -2.05 to 1.25). The global symptom scores (whether individuals felt better or worse) were reported in the paper to favour cisapride and be statistically significant (P < 0.05).We assessed the available data to be very low quality. There was a great deal of missing data from the included trial and the investigators failed to report numerical data for many outcomes. The overall risk of bias of the trial was unclear and it had a high risk for reporting bias. There was also indirectness; the trial drug (cisapride) has since been removed from the market in several countries due to adverse effects, thus it has no current applicability for preventing DIOS. The included trial also had very few participants, which downgraded the quality a further level for precision. There is an absence of evidence for interventions for the prevention of DIOS. As there was only one included trial, we could not perform a meta-analysis of the data. Furthermore, the included trial compared a prokinetic agent (cisapride) that is no longer licensed for use in a number of countries due to the risk of serious cardiac events, a finding that came to light after the trial was conducted. Therefore, the limited findings from the trial are not applicable in current clinical practice.Overall, a great deal more research needs to be undertaken on gastrointestinal complications in CF, as this is a very poorly studied area compared to respiratory complications in CF.


[Aldose reductase in the polyol pathway: a potential target for the therapeutic intervention of diabetic complications].
Aldose reductase (AR), an enzyme in the polyol pathway, catalyzes the reduction of glucose to sorbitol. Sorbitol is subsequently converted to fructose by sorbitol dehydrogenase. The two enzymes constitute the sorbitol (polyol) pathway, the alternate route of glucose metabolism. The acceleration of this pathway and ensuing metabolic imbalances have been postulated to play a key role in the pathogenesis of diabetic complications. Using a transgenic animal model expressing human AR, we defined the primary role of this pathway in the development of functional and structural abnormalities elicited by diabetes. The inhibitors for AR would thus become effective therapeutic agents for diabetic complications. As AR is a member of the structurally related, NADPH-dependent aldo-keto reductase superfamily, other members of this family, coexisting with AR, may interact with the inhibitors to quench their action against AR. With our new immunoassay system, the levels of AR expressed in diabetic patients can be measured directly. The enzyme levels were significantly associated with the presence of complications, indicating that variable levels of AR expressed in diabetic individuals may affect the susceptibility or development of pathological changes associated with diabetes. In this review, recent advances in the understanding of the pathophysiological significance of AR are presented that would aid in the effective pharmacological intervention of diabetic complications.


Outbreak of late-onset Group B Streptococcal disease with serotype Ib in a Neonatal Intensive Care Unit.
Hospital outbreaks of invasive group B streptococcus (GBS) infection are rare. There are only a few published reports of late-onset GBS outbreaks in neonatal intensive care units (NICUs). We report here three cases of late-onset GBS in our NICU. Three preterm very low birth weight (VLBW) infants born at 24 -27 weeks gestation developed lateonset GBS sepsis within four weeks. Two asymptomatic GBS carriers were identified in the NICU prior to the outbreak. Tests of maternal rectovaginal GBS colonization were negative in all three cases; as such, vertical transmission was unlikely. All three GBS isolates were capsular serotype 1b, with comparable antibiotic susceptibility profiles. Preterm delivery and VLBW are associated with an increased risk of invasive late-onset GBS infection. This report underscores the ongoing risk of nosocomial transmission of GBS in the NICU.


Antibacterial Activity of Terpenes and Terpenoids Present in Essential Oils.
Background: The antimicrobial activity of essential oils has been reported in hundreds of studies, however, the great majority of these studies attribute the activity to the most prevalent compounds without analyzing them independently. Therefore, the aim was to investigate the antibacterial activity of 33 free terpenes commonly found in essential oils and evaluate the cellular ultrastructure to verify possible damage to the cellular membrane. Methods: Screening was performed to select substances with possible antimicrobial activity, then the minimal inhibitory concentrations, bactericidal activity and 24-h time-kill curve studies were evaluated by standard protocols. In addition, the ultrastructure of control and death bacteria were evaluated by scanning electron microscopy. Results: Only 16 of the 33 compounds had antimicrobial activity at the initial screening. Eugenol exhibited rapid bactericidal action against Salmonella enterica serovar Typhimurium (2 h). Terpineol showed excellent bactericidal activity against S. aureus strains. Carveol, citronellol and geraniol presented a rapid bactericidal effect against E. coli. Conclusions: The higher antimicrobial activity was related to the presence of hydroxyl groups (phenolic and alcohol compounds), whereas hydrocarbons resulted in less activity. The first group, such as carvacrol, l-carveol, eugenol, trans-geraniol, and thymol, showed higher activity when compared to sulfanilamide. Images obtained by scanning electron microscopy indicate that the mechanism causing the cell death of the evaluated bacteria is based on the loss of cellular membrane integrity of function. The present study brings detailed knowledge about the antimicrobial activity of the individual compounds present in essential oils, that can provide a greater understanding for the future researches.


Expression of E6AP and PML predicts for prostate cancer progression and cancer-specific death.
The promyelocytic leukemia (PML) tumor suppressor plays an important role in the response to a variety of cellular stressors and its expression is downregulated or lost in a range of human tumors. We have previously shown that the E3 ligase E6-associated protein (E6AP) is an important regulator of PML protein stability but the relationship and clinical impact of PML and E6AP expression in prostatic carcinoma is unknown. E6AP and PML expression was assessed in tissue microarrays from a phase I discovery cohort of 170 patients treated by radical prostatectomy for localized prostate cancer (PC). Correlation analysis was carried out between PML and E6AP expression and clinicopathological variates including PSA as a surrogate of disease recurrence. The results were confirmed in a phase II validation cohort of 318 patients with associated clinical recurrence and survival data. Survival analysis of the phase I cohort revealed that patients whose tumors showed reduced PML and high E6AP expression had reduced time to PSA relapse (P = 0.012). This was confirmed in the phase II validation cohort where the expression profile of high E6AP/low PML was significantly associated with reduced time to PSA relapse (P < 0.001), clinical relapse (P = 0.016) and PC-specific death (P = 0.014). In multivariate analysis, this expression profile was an independent prognostic indicator of PSA relapse and clinical relapse independent of clinicopathologic factors predicting recurrence. This study identifies E6AP and PML as potential prognostic markers in localized prostate carcinoma and supports a role for E6AP in driving the downregulation or loss of PML expression in prostate carcinomas.


The effects of resveratrol on the healing of left colonic anastomosis.
Resveratrol (RSV) is a natural polyphenolic compound found in grape skins and the red wine which improves histological reorganization of the regenerating tissue in dermal wound healing. Since anastomotic healing possesses paramount importance to prevent complications in colorectal surgery, the present study is aimed to evaluate the effect of RSV on the healing of experimental left colonic anastomoses. Thirty-two male Wistar albino rats were randomized into two groups and subjected to colonic anastomosis. The study group was treated with RSV and the control group received tap water instead. The rats were sacrificed 3 and 7 days postoperatively. Wound complications, intra-abdominal abscesses, and anastomotic leaks and stenosis were recorded. Four types of assessment were performed: bursting pressure, hydroxyproline (OHP) content, histopathology, and biochemical analysis. Compared to the control group, the RSV-treated rats displayed a higher bursting pressure (p < .001) and anastomotic OHP content (p < .05)]. RSV treatment leads to significant increase in PON activity at both time points and decrease in malondialdehyde levels on postoperative day 3 (p < .001). Histopathological analysis revealed that RSV administration leads to a better anastomotic healing in terms of mucosal ischemia, neovascularization, reepithelialization, fibroblast, and lymphocyte infiltration. The study results suggest that exogenous RSV administration exerts a positive effect on experimental colonic wound healing in the rat. Although the precise cellular mechanisms by which RSV enhances anastomotic wound healing is not clear, stimulation of neovascularization, generation of collagen synthesis, inhibition of over inflammation, and restriction of oxidative injury seems to be of paramount importance.


Influence of impaired liver methionine metabolism on the development of vascular disease and inflammation.
Methionine (Met) metabolism involves the sequential formation of S-adenosylmethionine (SAM, the main biological methyl donor), S-adenosylhomocysteine (SAH) and homocysteine (Hcy). Hcy can be remethylated to Met or catabolized through the trans-sulfuration pathway. In mammals, as much as 48% of Met metabolism and up to 85% of all transmethylation reactions occur in the liver. These figures underscore the central role played by this organ in Met metabolism. Maintaining the homeostasis of this metabolic cycle has proved to be essential for the preservation of liver function up to the point of preventing its neoplastic transformation. However, an adequate hepatic metabolism of Met is not only important for the liver parenchymal cell. Evidence has accumulated over the past few years supporting the involvement of Met-derived metabolites in the triggering or attenuation of pathological processes with systemic implications. This is best illustrated by the fact that a deteriorated liver function has emerged as a major factor in the development of hyperhomocysteinemia. Elevated plasma levels of Hcy have been related to several disorders including cardiovascular and cerebrovascular diseases. On the other end, liver damage also leads to deficient SAM synthesis. Among the consequences of impaired SAM synthesis in liver tissue are the enhanced production of pro-inflammatory cytokines and mediators. In this review, we will address the mechanisms and consequences of abnormal Met metabolism in liver injury, the systemic implications of such impairment and finally the potential therapeutic interventions.


Comparing IV With Structural Models: What Simple IV Can and Cannot Identify.
This paper compares the economic questions addressed by instrumental variables estimators with those addressed by structural approaches. We discuss Marschak's Maxim: estimators should be selected on the basis of their ability to answer well-posed economic problems with minimal assumptions. A key identifying assumption that allows structural methods to be more informative than IV can be tested with data and does not have to be imposed.


A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death.
X chromosome-linked inhibitor of apoptosis (XIAP) is an endogenous inhibitor of caspase-3, -7, and -9. Smac/DIABLO, an inhibitor of XIAP, is released from mitochondria upon receiving apoptotic stimuli and binds to the BIR2 and BIR3 domains of XIAP, thereby inhibiting its caspase-inhibitory activity. Here we report that a serine protease called HtrA2/Omi is released from mitochondria and inhibits the function of XIAP by direct binding in a similar way to Smac. Moreover, when overexpressed extramitochondrially, HtrA2 induces atypical cell death, which is neither accompanied by a significant increase in caspase activity nor inhibited by caspase inhibitors, including XIAP. A catalytically inactive mutant of HtrA2, however, does not induce cell death. In short, HtrA2 is a Smac-like inhibitor of IAP activity with a serine protease-dependent cell death-inducing activity.


Couples' Time Together: Complementarities in Production versus Complementarities in Consumption.
Economists have previously suggested that gains from marriage can be generated by complementarities in production (gains from specialization and exchange) or by complementarities in consumption (gains from joint consumption of household public goods and joint time consumption). This paper uses the American Time Use Survey (ATUS) from 2003-2011 to test whether couples that engage in less specialization (are more similar in hours of market work) spend more time together. We find that among married couples without young children, those with a greater difference in weekly hours of work between husband and wife spend less time together on non-working weekend days. Importantly, we find that this relationship is quite symmetric between couples in which the husband works greater hours and couples in which the wife works greater hours. We do not find evidence of a relationship between specialization and couple time together among couples with young children.


The critical role of novel benzophenone analogs on tumor growth inhibition targeting angiogenesis and apoptosis.
In modern biology, one of the major topics of importance is progress in anti-cancer drugs with specific targets. The angiopreventive and in vitro tumor inhibition activities of novel synthetic benzophenone analogs have been investigated intensively and explored in a very systematic way. Novel benzophenone analogs (9a-d and 10a-d) substituted with methyl, chloro and fluoro groups at different positions on an identical chemical backbone and incorporating variations in the number of substituents have been synthesized in a multistep process and characterized. In this study, we further evaluate the newly synthesized compounds for their cytotoxic and anti-proliferative effects against A549, HeLa and MCF-7 cells. The potent lead compound was further assessed for anti-angiogenic effects. Through the structure-activity relationship, we found that an increase in the number of methyl, chloro and fluoro groups in a benzophenone ring on compound 9d resulted in higher potency compared to other compounds. Tumor inhibition was notably promoted, and this was reflected in effects on neovessel formation in in vivo systems, such as the CAM. Compound 9d interacts with rVEGF through hydrogen bonds in silico, thereby down-regulating the expression of VEGF in angiogenesis. From our investigation, it is suggested on the basis of clonogenesis and cell migration assays that compound 9d has the potency to exhibit prolonged activity against cancer progression, through cell cycle arrest at the G2/M phase. In addition, compound 9d inhibits A549 cells through caspase-activated DNase-mediated apoptosis.


Unbiased proteomics analysis demonstrates significant variability in mucosal immune factor expression depending on the site and method of collection.
Female genital tract secretions are commonly sampled by lavage of the ectocervix and vaginal vault or via a sponge inserted into the endocervix for evaluating inflammation status and immune factors critical for HIV microbicide and vaccine studies. This study uses a proteomics approach to comprehensively compare the efficacy of these methods, which sample from different compartments of the female genital tract, for the collection of immune factors. Matching sponge and lavage samples were collected from 10 healthy women and were analyzed by tandem mass spectrometry. Data was analyzed by a combination of differential protein expression analysis, hierarchical clustering and pathway analysis. Of the 385 proteins identified, endocervical sponge samples collected nearly twice as many unique proteins as cervicovaginal lavage (111 vs. 61) with 55% of proteins common to both (213). Each method/site identified 73 unique proteins that have roles in host immunity according to their gene ontology. Sponge samples enriched for specific inflammation pathways including acute phase response proteins (p = 3.37×10(-24)) and LXR/RXR immune activation pathways (p = 8.82×10(-22)) while the role IL-17A in psoriasis pathway (p = 5.98×10(-4)) and the complement system pathway (p = 3.91×10(-3)) were enriched in lavage samples. Many host defense factors were differentially enriched (p<0.05) between sites including known/potential antimicrobial factors (n = 21), S100 proteins (n = 9), and immune regulatory factors such as serpins (n = 7). Immunoglobulins (n = 6) were collected at comparable levels in abundance in each site although 25% of those identified were unique to sponge samples. This study demonstrates significant differences in types and quantities of immune factors and inflammation pathways collected by each sampling technique. Therefore, clinical studies that measure mucosal immune activation or factors assessing HIV transmission should utilize both collection methods to obtain the greatest representation of immune factors secreted into the female genital tract.


Molecular and morphological characterisation of Pseudococcidae surveyed on crops and ornamental plants in Spain.
Mealybugs (Hemiptera: Pseudococcidae) are common invasive pests in Europe, causing major problems on crops and ornamental plants. However, very few data are available concerning the mealybug fauna of southern Europe. This lack of data and the difficulty of identifying mealybugs morphologically by traditional techniques currently limit the perspectives for efficient specific pest management. The aim of this study was to provide multi-criterion characterization of mealybugs surveyed in eastern Spain in order to facilitate their routine identification through DNA sequencing or the use of derived species-specific molecular tools. We characterised 33 mealybug populations infesting crops and ornamental plants in eastern Spain, using a combination of molecular and morphological techniques, including the sequencing of the universal barcode DNA region cytochrome c oxidase subunit I (COI). This characterisation has led to the identification of ten species and provides sequence data for three previously unsequenced species, contributing to the phylogenetic knowledge of the family Pseudococcidae. In addition, the intraspecific variations found in the populations of five mealybug species provide insight into their invasion history.


Inhibition of allergen-induced airway obstruction and leukotriene generation in atopic asthmatic subjects by the leukotriene biosynthesis inhibitor BAYx 1005.
Leukotriene receptor antagonists significantly blunt allergen-induced bronchoconstriction in asthmatic subjects. Inhibitors of leukotriene synthesis should theoretically provide similar protection, but conflicting results have been obtained when synthesis inhibitors have been tested in allergen challenge. BAYx 1005, a new inhibitor of leukotriene synthesis, was therefore evaluated in an allergen bronchoprovocation study. Ten men with mild allergic asthma and bronchial hyperresponsiveness to histamine were recruited. On two different occasions each subject inhaled a single dose of allergen, previously determined to cause at least a 20% fall in forced expiratory volume in one second (FEV1) four hours after ingestion of 750 mg BAYx 1005 or placebo in a double blind crossover design. Urinary excretion of leukotriene E4 was measured before and during the challenges. The mean (SE) maximal fall in FEV1 was 7.1 (1.7)% after BAYx 1005 and 21.0 (3.0)% after placebo (p < 0.001). The mean difference between treatments was 13.9 (95% CI 7.0 to 20.8) for the maximal fall in FEV1. All subjects were protected by BAYx 1005, the mean inhibition of the fall in FEV1 being 70.0 (7.0)%. The mean area under the curve (AUC) for urinary excretion of leukotriene E4 in the first two hours after the challenge was 1.7 (0.9) after placebo and 0.4 (0.6) after BAYx 1005 (difference = 1.3 (95% CI-0.1 to 2.7); p < 0.05). These results indicate that BAYx 1005 is a potent inhibitor of allergen-provoked leukotriene synthesis in asthmatic subjects and lend further support to the suggestion that leukotrienes are important mediators of allergen-induced bronchoconstriction.


Exercise training in patients with a left ventricular assist device (Ex-VAD): rationale and design of a multicentre, prospective, assessor-blinded, randomized, controlled trial.
Left ventricular assist device (LVAD) therapy is a promising option for patients with advanced heart failure (HF), refractory to guideline-mandated medical treatment either as a bridge to heart transplantation or as lifelong therapy. Functional capacity improves after LVAD implantation but remains reduced in patients with long-term LVAD therapy. Exercise training (ET) improves functional capacity and quality of life (QoL) in HF and may provide incremental benefits in patients supported with LVAD therapy. The primary objective of Ex-VAD is to investigate whether a 12-week supervised ET can improve peak oxygen uptake (peakVO2 ) measured by cardiopulmonary exercise testing (CPET) on an ergometer. The study is powered to demonstrate a group difference of 3 mL/min/kg in peakVO2 at week 12, with a power of 0.9 and a standard deviation of 5 mL/min/kg. After baseline assessments to determine whether ET is safe, 66 patients at six trial sites with advanced HF and LVAD therapy will be randomized 2:1 to supervised ET or to the control arm of usual care alone. Patients randomized to ET will perform supervised aerobic endurance and resistance ET (three times/week) for 12 weeks. At baseline and during follow-up, anthropometry, CPET, echocardiography (at rest and exercise), and QoL evaluation will be performed. Blood samples will be collected to examine cardiac-specific relevant biomarkers. Overall physical activity, training sessions, and adherence will be monitored and documented throughout the study using accelerometers and patient diaries. The Ex-VAD trial will assess the effects of a supervised ET programme on peakVO2 and QoL in patients with LVAD. As LVAD therapy moves from crisis support to ambulatory functional enhancement, this trial will provide a rationale to improve functional capacity and, in perspective, cardiovascular outcomes in LVAD-supported patients with advanced HF.


An integrated optical modulator operating at cryogenic temperatures.
Photonic integrated circuits (PICs) operating at cryogenic temperatures are fundamental building blocks required to achieve scalable quantum computing and cryogenic computing technologies1,2. Silicon PICs have matured for room-temperature applications, but their cryogenic performance is limited by the absence of efficient low-temperature electro-optic modulation. Here we demonstrate electro-optic switching and modulation from room temperature down to 4 K by using the Pockels effect in integrated barium titanate (BaTiO3) devices3. We investigate the temperature dependence of the nonlinear optical properties of BaTiO3, showing an effective Pockels coefficient of 200 pm V-1 at 4 K. The fabricated devices show an electro-optic bandwidth of 30 GHz, ultralow-power tuning that is 109 times more efficient than thermal tuning, and high-speed data modulation at 20 Gbps. Our results demonstrate a missing component for cryogenic PICs, removing major roadblocks for the realization of cryogenic-compatible systems in the field of quantum computing, supercomputing and sensing, and for interfacing those systems with instrumentation at room temperature.


Exploring mobile working in healthcare: Clinical perspectives on transitioning to a mobile first culture of work.
This study investigated the views of healthcare professionals (HCPs) as end-users of mobile technologies to inform the requirements for a successful move towards a mobile first culture of work within secondary care. Many HCPs already used mobile devices to support their day-to-day practice and so transitioning to an organisational led mobile way of working is both needed and timely. In-depth focus groups and interviews at a UK academic hospital were conducted with HCPs (nurses, doctors, and allied health professionals). The interviews elicited views from HCPs about the use of mobile technologies in their clinical work and discussed their experiences of previous technology deployments. Thirty-four HCPs participated. Three themes were identified: integrating mobile working in hospital care; addressing data governance and accountability for mobile working; and handling the pace of change. The acceptance of mobile working is determined by whether it enhances, rather than hinders clinical practice whilst holding end-user's confidence; measuring the impact of the administrative burden on workload, changes to professional accountability and safeguarding patient data are serious issues that need consideration; and the pace of change needs to work for all stakeholders. The benefits of adopting mobile technologies are well recognised, however, technical and policy related factors identified by professional end-users require answers in order to successfully integrate mobile working into current practice. Adopting a mobile first approach to work is timely and achievable, but can only be delivered on a foundation of positive end-user engagement and strong organisational leadership.


Management and treatment of lactose malabsorption.
Lactose malabsorption is a very common condition characterized by intestinal lactase deficiency. Primary lactose malabsorption is an inherited deficit present in the majority of the world's population, while secondary hypolactasia can be the consequence of an intestinal disease. The presence of malabsorbed lactose in the colonic lumen causes gastrointestinal symptoms. The condition is known as lactose intolerance. In patients with lactase nonpersistence, treatment should be considered exclusively if intolerance symptoms are present. In the absence of guidelines, the common therapeutic approach tends to exclude milk and dairy products from the diet. However, this strategy may have serious nutritional disadvantages. Several studies have been carried out to find alternative approaches, such as exogenous beta-galactosidase, yogurt and probiotics for their bacterial lactase activity, pharmacological and non pharmacological strategies that can prolong contact time between enzyme and substrate delaying gastrointestinal transit time, and chronic lactose ingestion to enhance colonic adaptation. In this review the usefulness of these approaches is discussed and a therapeutic management with a flow chart is proposed.


Isolation of intact, functional mitochondria from the model plant Arabidopsis thaliana.
The ability to isolate intact, functional mitochondria from plant tissues is a key technique in the study of the genome, proteome, and metabolic function of the plant mitochondrion. Traditionally, mitochondrial plant researchers have turned to specific plant systems and organs (such as potato tubers and pea shoots) from which mitochondria are readily isolated in large quantities. However, increasingly, research is focused on a small number of model species, and there is a need to adapt existing protocols to allow the isolation of mitochondria from these model species. Arguably, the most important of these is Arabidopsis thaliana, for which a formidable array of genetic resources is available. However, because of its relatively small size and the absence of large heterotrophic organs, Arabidopsis is a challenging plant from which to isolate mitochondria. Here, we present two methods for isolating mitochondria from Arabidopsis, either from heterotrophic cell suspension cultures or from hydroponic seedling cultures. We also present details of commonly used assays to assess the physical and functional integrity of the isolated organelles.


Persistent respiratory health effects after a metam sodium pesticide spill.
To report the occurrence of persistent respiratory disorders, including irritant-induced asthma, among adults living and working near an environmental spill of the pesticide, metam sodium, after the derailment of a tank car. Retrospective clinical case series. California communities situated within one-half mile of the Sacramento River, from Mt. Shasta City to Shasta Lake. 197 adults referred to a university occupational/environmental health clinic or to a private occupational/environmental health practitioner for evaluation of health problems potentially related to the spill. History, physical examination, review of medical records, spirometry, and methacholine challenge testing revealed 20 cases of persistent irritant-induced asthma and 10 cases of persistent exacerbation of asthma. This is the first reported series of cases of persistent irritant-induced asthma involving both community residents and occupationally exposed individuals.


Heteronuclear decoupling interference during symmetry-based homonuclear recoupling in solid-state NMR.
We examine the influence of continuous-wave heteronuclear decoupling on symmetry-based double-quantum homonuclear dipolar recoupling, using experimental measurements, numerical simulations, and average Hamiltonian theory. There are two distinct regimes in which the heteronuclear interference effects are minimized. The first regime utilizes a moderate homonuclear recoupling field and a strong heteronuclear decoupling field; the second regime utilizes a strong homonuclear recoupling field and a weak or absent heteronuclear decoupling field. The second regime is experimentally accessible at moderate or high magic-angle-spinning frequencies and is particularly relevant for many realistic applications of solid-state NMR recoupling experiments to organic or biological materials.


Multifunctional carbon nanoelectrodes fabricated by focused ion beam milling.
We report a strategy for fabrication of sub-micron, multifunctional carbon electrodes and application of these electrodes as probes for scanning electrochemical microscopy (SECM) and scanning ion conductance microscopy (SICM). The fabrication process utilized chemical vapor deposition of parylene, followed by thermal pyrolysis to form conductive carbon and then further deposition of parylene to form an insulation layer. To achieve well-defined electrode geometries, two methods of electrode exposure were utilized. In the first method, carbon probes were masked in polydimethylsiloxane (PDMS) to obtain a cone-shaped electrode. In the second method, the electrode area was exposed via milling with a focused ion beam (FIB) to reveal a carbon ring electrode, carbon ring/platinum disk electrode, or carbon ring/nanopore electrode. Carbon electrodes were batch fabricated (~35/batch) through the vapor deposition process and were characterized with scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM), and cyclic voltammetry (CV) measurements. Additionally, Raman spectroscopy was utilized to examine the effects of Ga(+) ion implantation, a result of FIB milling. Constant-height, feedback mode SECM was performed with conical carbon electrodes and carbon ring electrodes. We demonstrate the utility of carbon ring/nanopore electrodes with SECM-SICM to simultaneously collect topography, ion current and electrochemical current images. In addition, carbon ring/nanopore electrodes were utilized in substrate generation/tip collection (SG/TC) SECM. In SG/TC SECM, localized delivery of redox molecules affords a higher resolution, than when the redox molecules are present in the bath solution. Multifunctional geometries of carbon electrode probes will find utility in electroanalytical applications, in general, and more specifically with electrochemical microscopy as discussed herein.


Anti-inflammatory and analgesic activities of a novel biflavonoid from shells of Camellia oleifera.
Shells are by-products of oil production from Camellia oleifera which have not been harnessed effectively. The purpose of this research is to isolate flavonoid from shells of Camellia oleifera and evaluate its anti-inflammatory and analgesic effects. The flavonoid was identified as bimolecular kaempferol structure by UV, MS, 1H NMR and 13C NMR spectra, which is a new biflavonoid and first found in Camellia oleifera. It showed dose-dependent anti-inflammatory activity by carrageenin-induced paw oedema in rats and croton oil induced ear inflammation in mice, and analgesic activity by hot plate test and acetic acid induced writhing. The mechanism of anti-inflammation of biflavonoid is related to both bradykinin and prostaglandins synthesis inhibition. The biflavonoid showed both central and peripheral analgesic effects different from aspirin, inhibition of the synthesis or action of prostaglandins may contribute to analgesic effect of biflavonoid. The biflavonoid significantly decreased malonaldehyde (MDA) and increased superoxidase dismutase (SOD) and Glutathione peroxidase (GSH-Px) activity in serum (p < 0.01), revealed strong free radical scavenging activity in vivo. It indicates the biflavonoid can control inflammation and pain by eliminating free radical so as to inhibit the mediators and decrease the prostaglandins. The biflavonoid can be used as a prospective medicine for inflammation and pain.


Species interactions in mixed-community crystalline biofilms on urinary catheters.
Previous experimental investigations of the crystalline biofilms that colonize and block urinary catheters have focussed on their formation by pure cultures of Proteus mirabilis. In the urine of patients undergoing long-term catheterization, P. mirabilis is commonly found in mixed communities with other urinary tract pathogens. Little is known about the effect that the other species have on the rate at which P. mirabilis encrusts catheters. In the present study, a set of data on the nature of the bacterial communities on 106 catheter biofilms has been analysed and it was found that while species such as Providencia stuartii and Klebsiella pneumoniae were commonly associated with P. mirabilis, when Escherichia coli, Morganella morganii or Enterobacter cloacae were present, P. mirabilis was rarely or never found. The hypothesis that the absence of P. mirabilis from some biofilm communities could be due to its active exclusion by other species has also been examined. Experiments in laboratory models showed that co-infection of P. mirabilis with M. morganii, K. pneumoniae or E. coli had no effect on the ability of P. mirabilis to encrust and block catheters. Co-infection with Ent. cloacae or Pseudomonas aeruginosa, however, significantly increased the time that catheters took to block (P <0.05). The growth of Ent. cloacae, M. morganii, K. pneumoniae or E. coli in the model for 72 h prior to superinfection with P. mirabilis significantly delayed catheter blockage. In the case of Ent. cloacae, for example, the mean time to blockage was extended from 28.7 h to 60.7 h (P < or =0.01). In all cases, however, P. mirabilis was able to generate alkaline urine, colonize the biofilms, induce crystal formation and block the catheters. The results suggest that although there is a degree of antagonism between P. mirabilis and some of the other urinary tract organisms, the effects are temporary and whatever the pre-existing urinary microbiota, infection with P. mirabilis is thus likely to lead to catheter encrustation and blockage.


Benzodiazepine Use During Hospitalization: Automated Identification of Potential Medication Errors and Systematic Assessment of Preventable Adverse Events.
Benzodiazepines and "Z-drug" GABA-receptor modulators (BDZ) are among the most frequently used drugs in hospitals. Adverse drug events (ADE) associated with BDZ can be the result of preventable medication errors (ME) related to dosing, drug interactions and comorbidities. The present study evaluated inpatient use of BDZ and related ME and ADE. We conducted an observational study within a pharmacoepidemiological database derived from the clinical information system of a tertiary care hospital. We developed algorithms that identified dosing errors and interacting comedication for all administered BDZ. Associated ADE and risk factors were validated in medical records. Among 53,081 patients contributing 495,813 patient-days BDZ were administered to 25,626 patients (48.3%) on 115,150 patient-days (23.2%). We identified 3,372 patient-days (2.9%) with comedication that inhibits BDZ metabolism, and 1,197 (1.0%) with lorazepam administration in severe renal impairment. After validation we classified 134, 56, 12, and 3 cases involving lorazepam, zolpidem, midazolam and triazolam, respectively, as clinically relevant ME. Among those there were 23 cases with associated adverse drug events, including severe CNS-depression, falls with subsequent injuries and severe dyspnea. Causality for BDZ was formally assessed as 'possible' or 'probable' in 20 of those cases. Four cases with ME and associated severe ADE required administration of the BDZ antagonist flumazenil. BDZ use was remarkably high in the studied setting, frequently involved potential ME related to dosing, co-medication and comorbidities, and rarely cases with associated ADE. We propose the implementation of automated ME screening and validation for the prevention of BDZ-related ADE.