Heparin-induced thrombocytopenia (HIT) causing portosplenic, superior mesenteric, and splenic vein thrombosis resulting in splenic rupture and pulmonary emboli formation.
Heparin-induced thrombocytopenia (HIT) is a life-threatening complication of heparin administration. Of the few reported cases of HIT-associated intra-abdominal thrombosis, none to our knowledge provide multidetector-row computed tomography (MDCT) imaging findings or emphasize its utility in diagnosis. We describe a case of HIT with MDCT images demonstrating extensive intra-abdominal thrombosis and end-organ complications including splenic rupture and pulmonary emboli. This case emphasizes the potential role of MDCT in the rapid detection of HIT-related thromboembolic complications in patients with nonspecific abdominal pain.


Mass fatality preparedness among medical examiners/coroners in the United States: a cross-sectional study.
In the United States (US), Medical Examiners and Coroners (ME/Cs) have the legal authority for the management of mass fatality incidents (MFI). Yet, preparedness and operational capabilities in this sector remain largely unknown. The purpose of this study was twofold; first, to identify appropriate measures of preparedness, and second, to assess preparedness levels and factors significantly associated with preparedness. Three separate checklists were developed to measure different aspects of preparedness: MFI Plan Elements, Operational Capabilities, and Pre-existing Resource Networks. Using a cross-sectional study design, data on these and other variables of interest were collected in 2014 from a national convenience sample of ME/C using an internet-based, anonymous survey. Preparedness levels were determined and compared across Federal Regions and in relation to the number of Presidential Disaster Declarations, also by Federal Region. Bivariate logistic and multivariable models estimated the associations between organizational characteristics and relative preparedness. A large proportion (42%) of respondents reported that less than 25 additional fatalities over a 48-hour period would exceed their response capacities. The preparedness constructs measured three related, yet distinct, aspects of preparedness, with scores highly variable and generally suboptimal. Median scores for the three preparedness measures also varied across Federal Regions and as compared to the number of Presidential Declared Disasters, also by Federal Region. Capacity was especially limited for activating missing persons call centers, launching public communications, especially via social media, and identifying temporary interment sites. The provision of staff training was the only factor studied that was significantly (positively) associated (p < .05) with all three preparedness measures. Although ME/Cs ranked local partners, such as Offices of Emergency Management, first responders, and funeral homes, as the most important sources of assistance, a sizeable proportion (72%) expected federal assistance. The three measures of MFI preparedness allowed for a broad and comprehensive assessment of preparedness. In the future, these measures can serve as useful benchmarks or criteria for assessing ME/Cs preparedness. The study findings suggest multiple opportunities for improvement, including the development and implementation of national strategies to ensure uniform standards for MFI management across all jurisdictions.


Beyond the Trinity of ATM, ATR, and DNA-PK: Multiple Kinases Shape the DNA Damage Response in Concert With RNA Metabolism.
Our genome is constantly exposed to endogenous and exogenous sources of DNA damage resulting in various alterations of the genetic code. DNA double-strand breaks (DSBs) are considered one of the most cytotoxic lesions. Several types of repair pathways act to repair DNA damage and maintain genome stability. In the canonical DNA damage response (DDR) DSBs are recognized by the sensing kinases Ataxia-telangiectasia mutated (ATM), Ataxia-telangiectasia and Rad3-related (ATR), and DNA-dependent protein kinase (DNA-PK), which initiate a cascade of kinase-dependent amplification steps known as DSB signaling. Recent evidence suggests that efficient recognition and repair of DSBs relies on the transcription and processing of non-coding (nc)RNA molecules by RNA polymerase II (RNAPII) and the RNA interference (RNAi) factors Drosha and Dicer. Multiple kinases influence the phosphorylation status of both the RNAPII carboxy-terminal domain (CTD) and Dicer in order to regulate RNA-dependent DSBs repair. The importance of kinase signaling and RNA processing in the DDR is highlighted by the regulation of p53-binding protein (53BP1), a key regulator of DSB repair pathway choice between homologous recombination (HR) and non-homologous end joining (NHEJ). Additionally, emerging evidence suggests that RNA metabolic enzymes also play a role in the repair of other types of DNA damage, including the DDR to ultraviolet radiation (UVR). RNAi factors are also substrates for mitogen-activated protein kinase (MAPK) signaling and mediate the turnover of ncRNA during nucleotide excision repair (NER) in response to UVR. Here, we review kinase-dependent phosphorylation events on RNAPII, Drosha and Dicer, and 53BP1 that modulate the key steps of the DDR to DSBs and UVR, suggesting an intimate link between the DDR and RNA metabolism.


Color contrast processing in human striate cortex.
Color constancy refers to the unchanging nature of the perceived color of an object despite considerable variation in the wavelength composition of the light illuminating it. The color contrasts between objects and their backgrounds play a crucial role in color constancy. We tested a patient whose right striate cortex had been removed and demonstrated that he made no use of color contrast in judging color appearance but instead made judgments based simply on wavelength comparison. This was shown by presenting pairs of colored stimuli against a background color that gradually changed across space. When presented with such displays, both normal observers and those with cerebral achromatopsia (cortical color blindness) judge the color appearance of such stimuli on the basis of the chromatic contrast the stimuli make against their background rather than on the physical wavelengths of the light emitted from them. However, our patient made no such use of color contrast but, instead, made color discriminations simply on the basis of wavelength composition. This is consistent with recent findings from monkey electrophysiology that identify cells in early cortical visual areas that signal local contrast and so contribute to the likely mechanism for achieving color constancy.


Avipoxviruses: infection biology and their use as vaccine vectors.
Avipoxviruses (APVs) belong to the Chordopoxvirinae subfamily of the Poxviridae family. APVs are distributed worldwide and cause disease in domestic, pet and wild birds of many species. APVs are transmitted by aerosols and biting insects, particularly mosquitoes and arthropods and are usually named after the bird species from which they were originally isolated. The virus species Fowlpox virus (FWPV) causes disease in poultry and associated mortality is usually low, but in flocks under stress (other diseases, high production) mortality can reach up to 50%. APVs are also major players in viral vaccine vector development for diseases in human and veterinary medicine. Abortive infection in mammalian cells (no production of progeny viruses) and their ability to accommodate multiple gene inserts are some of the characteristics that make APVs promising vaccine vectors. Although abortive infection in mammalian cells conceivably represents a major vaccine bio-safety advantage, molecular mechanisms restricting APVs to certain hosts are not yet fully understood. This review summarizes the current knowledge relating to APVs, including classification, morphogenesis, host-virus interactions, diagnostics and disease, and also highlights the use of APVs as recombinant vaccine vectors.


Proteomic analysis of rhoptry organelles reveals many novel constituents for host-parasite interactions in Toxoplasma gondii.
Rhoptries are specialized secretory organelles that are uniquely present within protozoan parasites of the phylum Apicomplexa. These obligate intracellular parasites comprise some of the most important parasites of humans and animals, including the causative agents of malaria (Plasmodium spp.) and chicken coccidiosis (Eimeria spp.). The contents of the rhoptries are released into the nascent parasitophorous vacuole during invasion into the host cell, and the resulting proteins often represent the literal interface between host and pathogen. We have developed a method for highly efficient purification of rhoptries from one of the best studied Apicomplexa, Toxoplasma gondii, and we carried out a detailed proteomic analysis using mass spectrometry that has identified 38 novel proteins. To confirm their rhoptry origin, antibodies were raised to synthetic peptides and/or recombinant protein. Eleven of 12 of these yielded antibody that showed strong rhoptry staining by immunofluorescence within the rhoptry necks and/or their bulbous base. Hemagglutinin epitope tagging confirmed one additional novel protein as from the rhoptry bulb. Previously identified rhoptry proteins from Toxoplasma and Plasmodium were unique to one or the other organism, but our elucidation of the Toxoplasma rhoptry proteome revealed homologues that are common to both. This study also identified the first Toxoplasma genes encoding rhoptry neck proteins, which we named RONs, demonstrated that toxofilin and Rab11 are rhoptry proteins, and identified novel kinases, phosphatases, and proteases that are likely to play a key role in the ability of the parasite to invade and co-opt the host cell for its own survival and growth.


Synthesis of spatial coherence.
The mutual intensity function plays a major role in characterizing quasi-monochromatic, partially coherent optical signals. We propose to use the mutual intensity as a carrier of information to avoid speckle noise in coherent illumination systems and to permit the use of complex functions that are prohibited spatially incoherent sources. To do this we require methods for encoding the information as a coherence function. An optical system for synthesizing a beam with a given mutual intensity function is proposed. The optical system permits the synthesis of any desired mutual intensity function. The illumination is supplied by a quasi-monochromatic, spatially incoherent source. Experimental results demonstrate the performance of this system for several cases.


Lung health among plumbers and pipefitters in Edmonton, Alberta.
A cross sectional study was undertaken to assess lung health among plumbers and pipefitters. Respiratory symptoms, lung function, and radiographic changes among 99 actively employed plumbers and pipefitters with > or = 20 years of union membership were compared with 100 telephone workers. A respiratory symptom questionnaire was administered, including smoking and occupational histories. Spirometry was conducted according to standard criteria. Posteroanterior chest radiographs were evaluated by two experienced chest physicians, with a third arbitrating disagreed films. Members of the union were categorised as pipefitters (n = 57), plumbers (n = 16), or welders (n = 26), based on longest service, and compared with the telephone workers and internally (between groups). Lung health was also compared with employment in several work sectors common to Alberta for time, and for time weighted by exposure to dust and fumes. Compared with the telephone workers, plumbers and pipefitters had more cough and phlegm, lower forced vital capacity, and more radiographic changes (20% with any change), including circumscribed (10%) and diffuse pleural thickening (9%). None of the plumbers and pipefitters had small radiographic opacities. Among the three subgroups of workers, plumbers had the highest prevalence of radiographic changes. Both plumbers and pipefitters showed higher odds ratios for cough and phlegm than the welders. No differences between groups were found for lung function. Indicators of lung health were not related to work in any sector. Plumbers and pipefitters had increased prevalence of symptoms suggestive of an irritant effect with no evidence of bronchial responsiveness. The chest radiographs showed evidence of asbestos exposure, especially in the plumbers, but at lower levels than previously reported. Health screening programmes for these workers should be considered, although the logistical problems associated with screening in this group would be considerable.


Grandparental and parental obesity influences on childhood overweight: implications for primary care practice.
Community-based studies have suggested a multigenerational pattern of obesity affecting children's risk of overweight, but no national data have substantiated such a pattern. Our objective was to examine the prevalence of overweight [body mass index (BMI) >or=95th percentile for age and sex] among children aged 5 to 19 in a national sample, stratified by the obesity status of their parents and grandparents. We used a secondary analysis of the Panel Study of Income Dynamics, Child Development Supplement, a multigenerational, genealogical, prospective cohort study of the US population. Self-report height and weight data from adults and measured height and weight data for children were used to calculate BMI. The prevalence of child overweight was calculated for different possible combinations of parental and grandparental BMI status, including missing status. The sample included 2,591 children aged 5 to 19 years, for whom parental BMI data were available for 94% and grandparental BMI data were available for 61%. Prevalence of childhood overweight (18.6%) in the sample was comparable with contemporaneous measured national data from other sources. Among children with normal-weight parents and normal-weight grandparents, 7.9% were overweight. In contrast, among children with overweight parents (BMI 25-29.9) and normal-weight grandparents, 17.9% were overweight, and among children with obese parents (BMI >or=30) and normal-weight grandparents, 31.9% were overweight (P < .0001). Importantly, when parents were normal weight, if grandparents were obese, then the prevalence of child overweight was 17.4% (P < .0001). The prevalence of child overweight was similarly elevated (16.4%) when parents were normal weight and grandparental BMI was missing. This is the first national study to find an association of child weight status with grandparental obesity, distinct from parental obesity. Primary care physicians may find it helpful to consider grandparents' weight status in judging risk of childhood overweight for their patients, especially when parents' weight is normal.


The Effect of Acute L-carnitine and Carbohydrate Intake on Cycling Performance.
Chronic supplementation of L-carnitine and carbohydrate has been reported to increase L-carnitine content in skeletal muscle and have positive influences on exercise variables and performance. This study investigated the acute intake of L-carnitine and carbohydrate on the exercise parameters of cycling. A total of 10 males (27.0 ± 4 years) completed two exercise sessions consisting of 40 min of cycling at 65% of VO2peak, followed by cycling to exhaustion at 85% of VO2peak. L-carnitine or a placebo was consumed 3 hours prior to exercise, and beverages consisting of 94 g of carbohydrate were consumed at both 2 hours, and 30 minutes prior to exercise. Repeated measures ANOVAs were used to compare respiratory exchange ratio (RER), blood lactate, and power output across experimental trials and time. A repeated measures t-test was used to analyze differences between conditions and time to exhaustion. RER was significantly lower (p=0.01) at baseline with L-carnitine ingestion (.83 ± .05) compared to the placebo ingestion (.86 ± .06). Blood lactate was significantly lower (p=0.02) after 10 minutes of cycling at 65% of VO2peak with ingestion of L-carnitine (35% change from baseline) compared to placebo ingestion (53% change from baseline). No differences were found for power output or time to exhaustion at 85% of VO2peak. Despite mentioned differences, acute intake of L-carnitine and carbohydrate does not appear to influence exercise parameters, likely due to a lack of sufficient change in the content of L-carnitine in skeletal muscle.


Genomic saturation mutagenesis and polygenic analysis identify novel yeast genes affecting ethyl acetate production, a non-selectable polygenic trait.
Isolation of mutants in populations of microorganisms has been a valuable tool in experimental genetics for decades. The main disadvantage, however, is the inability of isolating mutants in non-selectable polygenic traits. Most traits of organisms, however, are non-selectable and polygenic, including industrially important properties of microorganisms. The advent of powerful technologies for polygenic analysis of complex traits has allowed simultaneous identification of multiple causative mutations among many thousands of irrelevant mutations. We now show that this also applies to haploid strains of which the genome has been loaded with induced mutations so as to affect as many non-selectable, polygenic traits as possible. We have introduced about 900 mutations into single haploid yeast strains using multiple rounds of EMS mutagenesis, while maintaining the mating capacity required for genetic mapping. We screened the strains for defects in flavor production, an important non-selectable, polygenic trait in yeast alcoholic beverage production. A haploid strain with multiple induced mutations showing reduced ethyl acetate production in semi-anaerobic fermentation, was selected and the underlying quantitative trait loci (QTLs) were mapped using pooled-segregant whole-genome sequence analysis after crossing with an unrelated haploid strain. Reciprocal hemizygosity analysis and allele exchange identified PMA1 and CEM1 as causative mutant alleles and TPS1 as a causative genetic background allele. The case of CEM1 revealed that relevant mutations without observable effect in the haploid strain with multiple induced mutations (in this case due to defective mitochondria) can be identified by polygenic analysis as long as the mutations have an effect in part of the segregants (in this case those that regained fully functional mitochondria). Our results show that genomic saturation mutagenesis combined with complex trait polygenic analysis could be used successfully to identify causative alleles underlying many non-selectable, polygenic traits in small collections of haploid strains with multiple induced mutations.


Imaging of HIV-1 envelope-induced virological synapse and signaling on synthetic lipid bilayers.
Human immunodeficiency virus type 1 (HIV-1) infection occurs most efficiently via cell to cell transmission(2,10,11). This cell to cell transfer between CD4(+) T cells involves the formation of a virological synapse (VS), which is an F-actin-dependent cell-cell junction formed upon the engagement of HIV-1 envelope gp120 on the infected cell with CD4 and the chemokine receptor (CKR) CCR5 or CXCR4 on the target cell (8). In addition to gp120 and its receptors, other membrane proteins, particularly the adhesion molecule LFA-1 and its ligands, the ICAM family, play a major role in VS formation and virus transmission as they are present on the surface of virus-infected donor cells and target cells, as well as on the envelope of HIV-1 virions(1,4,5,6,7,13). VS formation is also accompanied by intracellular signaling events that are transduced as a result of gp120-engagement of its receptors. Indeed, we have recently showed that CD4(+) T cell interaction with gp120 induces recruitment and phosphorylation of signaling molecules associated with the TCR signalosome including Lck, CD3ζ, ZAP70, LAT, SLP-76, Itk, and PLCγ(15). In this article, we present a method to visualize supramolecular arrangement and membrane-proximal signaling events taking place during VS formation. We take advantage of the glass-supported planar bi-layer system as a reductionist model to represent the surface of HIV-infected cells bearing the viral envelope gp120 and the cellular adhesion molecule ICAM-1. The protocol describes general procedures for monitoring HIV-1 gp120-induced VS assembly and signal activation events that include i) bi-layer preparation and assembly in a flow cell, ii) injection of cells and immunofluorescence staining to detect intracellular signaling molecules on cells interacting with HIV-1 gp120 and ICAM-1 on bi-layers, iii) image acquisition by TIRF microscopy, and iv) data analysis. This system generates high-resolution images of VS interface beyond that achieved with the conventional cell-cell system as it allows detection of distinct clusters of individual molecular components of VS along with specific signaling molecules recruited to these sub-domains.


Sexual function in chronic illness.
Direct and indirect effects of chronic disease on sexual health are frequent and complex, but guidelines for their optimal management are lacking. With improved surgical and medical treatment of the underlying disease, the numbers of men and women needing assessment and management of associated sexual dysfunction are increasing. To provide recommendations/guidelines for the clinical management of sexual dysfunction within the context of chronic illness. An international consultation in collaboration with the major sexual medicine associations assembled 186 multidisciplinary experts from 33 countries into 25 committees. Nine experts from four countries compiled the recommendations of sexual dysfunction in chronic illness and cancer with four focusing on neurological, renal, and psychiatric disease and lower urinary tract symptoms (LUTS). Searches were conducted using Medline, Embase, Lilacs, and Pubmed databases. Expert opinion was based on grading of evidence-based medical literature, widespread internal committee discussion, public presentation, and debate. Some conclusions concerning prevalence and pathophysiology of sexual dysfunction in the context of neurological disorders, end-stage renal failure, LUTS, and psychiatric disease were made. Optimal assessment of the multiple factors affecting sexuality when one or both partners are chronically ill is outlined. Evidence-based recommendations for management are presented. Comorbid depression is frequent and independently determines prevalence of sexual dysfunction in many conditions. There is need for more research and scientific reporting on prevalence, pathophysiology, and optimal treatment of sexual dysfunction associated with chronic illness. Screening for and managing comorbid depression is strongly recommended.


Progestogens for Maintenance Tocolysis in Women With a Short Cervix: A Randomized Controlled Trial.
To assess the efficacy of progestogens for maintenance tocolysis in women undelivered after their first preterm labor episode. Women with singleton pregnancies between 22 0/7 and 31 6/7 weeks of gestation with arrested preterm labor and a cervical length 25 mm or less at hospital discharge were eligible. Patients with a previous preterm birth were excluded. In a randomized controlled trial conducted in five university hospitals, women were randomized to receive vaginal progesterone (200 mg per day) or intramuscular 17α-hydroxyprogesterone caproate (341 mg per week) or to an observation groups (control group). The primary outcome was the proportion of women with preterm birth at less than 37 weeks of gestation. A sample size of 160 per group (n=480) was planned to compare vaginal progesterone and 17α-hydroxyprogesterone caproate groups with those in the control group. The sample size estimation was based on the hypothesis that the risk of experiencing preterm birth in the control group would be 30% and that 17α-hydroxyprogesterone caproate or progesterone would decrease this risk to 15%. A P value of <.025 was defined as statistically significant. At planned interim analysis (n=254), the trial was stopped for futility. Between July 2010 and June 2015, 257 women were eligible and 254 were subsequently randomly assigned to vaginal progesterone (n=86), 17α-hydroxyprogesterone caproate (n=87), or observation (n=81). Nineteen (8%) were excluded from the analysis because they either dropped out or information was missing, leaving 235 women available for analysis. Demographic characteristics were similar across groups. The preterm birth rate did not differ significantly between groups: 23% in the 17α-hydroxyprogesterone caproate group, 39% in the vaginal progesterone group, and 22% in the women in the control group (P=.949 for 17α-hydroxyprogesterone caproate compared with the women in the control group and P=.027 for vaginal progesterone compared with women in the control group). The use of progestogens for maintenance tocolysis in women with a short cervix did not reduce the rate of preterm birth. ClinicalTrials.gov, NCT01178788.


Associations between heavy episodic drinking and alcohol related injuries: a case control study.
Alcohol is a significant risk factor for injuries. This study addresses 1) whether the risk of alcohol related injury increases with frequency of heavy episodic drinking (HED) in a linear fashion, and 2) whether a small group of high risk drinkers accounts for the majority of alcohol related injuries. We applied a case - control design. Cases were BAC positive injured patients (n = 534) and controls were respondents to a general population survey in Norway (n = 1947). Age and gender adjusted association between self-reported past year HED frequency and alcohol related injury risk was estimated in logistic regression models for all alcohol related injuries and for violence injuries and accident injuries separately. An increase in HED was associated with an increase in risk of alcohol related injury, resembling a linear risk function. The small fraction of high risk drinkers (6.6%) accounted for 41.6% of all alcohol related injuries, thus lending support to the validity of the prevention paradox. There is a strong relationship between frequency of heavy episodic drinking and risk of alcohol related injuries, yet the majority of alcohol related injuries are found among drinkers who are not in the high risk group.


Interaction between CD40 and its ligand gp39 in the development of murine lupus nephritis.
We investigated the role of gp39-CD40 interaction in the development of glomerulonephritis in lupus mice. In contrast to normal mice, lupus mice had much higher percentages of intensely gp39+ T cells in their spleens even at the preautoimmune age of 1 mo, and the further increase in gp39 expression by anti-CD3 Ab stimulation was markedly greater in lupus T cells. The pathogenic autoantibody-inducing ability of Th clones and splenic Th cells from lupus mice could be blocked in vitro by anti-gp39 Ab. Acceleration of lupus nephritis by the transfer of pathogenic autoantibody-inducing Th clones in vivo could also be completely blocked by anti-gp39 Ab. Surprisingly, a brief treatment of lupus mice with anti-gp39 Ab had a sustained beneficial effect on their spontaneous disease long after the Ab had been cleared from their systems. Only three injections of anti-gp39 Ab given to prenephritic lupus mice at 3 mo of age markedly delayed and reduced the incidence of lupus nephritis up to 12 mo of age by which time almost all the control mice had developed severe glomerulonephritis. Remarkably, pathogenic Th cells were left intact in these anti-gp39-treated mice but their B cells could not produce pathogenic autoantibodies even 9 mo after the therapy. Our studies suggest that blocking the interaction between gp39 on pathogenic Th cells and CD40 on lupus B cells at a crucial window of time delays the expansion autoimmune memory B cells resulting in long-term therapeutic benefits.


Attitudes of emergency department physicians and nurses toward implementation of an early warning score to identify critically ill patients: qualitative explanations for failed implementation.
Sepsis, a common, time-sensitive condition, is sometimes not identified at emergency department (ED) triage. The use of early warning scores has been shown to improve sepsis-related screening in other settings. Our objective was to elucidate nurse and physician perceptions with the Hamilton Early Warning Score (HEWS) in combination with the Canadian Triage Acuity Scale. Semi-structured interviews were conducted with nurses, resident physicians and attending physicians to explore perceived feasibility, utility, comfort, barriers, successes, opportunities and accuracy. A constructivist grounded theory approach was used. Transcripts were coded into thematic coding trees. The twelve participants did not value the HEWS in the ED because they felt it was not helpful in identifying critically ill patients. We identified five themes; knowledge of sepsis and HEWS, utility of HEWS in emergency triage, utility of HEWS at the bedside, utility in communicating acuity and deterioration, and feasibility and accuracy of data collection. We also found 9 barriers and 7 enablers to the use of early warning score in the ED. In our emergency departments, we identified potential barriers to implementation of an early warning score. A pre-existing expertise and lexicon related to critically ill patients lessens the perceived utility of an EWS in the ED. Understanding these cultural barriers needs to be addressed through change theory and implementation science.


In Silico and In Vitro Analysis of Major Cannabis-Derived Compounds as Fatty Acid Amide Hydrolase Inhibitors.
Accumulated evidence suggests that enhancing the endocannabinoid (eCB) tone, in particular of anandamide (N-arachidonoylethanolamine, AEA), has therapeutic potential in many human diseases. Fatty acid amide hydrolase (FAAH) is a membrane-bound enzyme principally responsible for the degradation of AEA, and thus it represents a relevant target to increase signaling thereof. In recent years, different synthetic and natural compounds have been developed and tested on rat FAAH, but little is known of their effect on the human enzyme. Here, we sought to investigate six major cannabis-derived compounds to compare their action on rat and human FAAHs. To this aim, we combined an in silico analysis of their binding mode and affinity, with in vitro assays of their effect on enzyme activity. This integrated approach allowed to disclose differences in efficacy towards rat and human FAAHs, and to highlight the role of key residues involved in the inhibition of both enzymes. This study suggests that the therapeutic efficacy of compounds targeted towards FAAH should be always tested in vitro on both rat and human enzymes.


A Longitudinal Examination of the Relationship between Trauma-Related Cognitive Factors and Internalising and Externalising Psychopathology in Physically Injured Children.
Cognitive models of posttraumatic stress disorder (PTSD) highlight maladaptive posttrauma appraisals, trauma memory qualities, and coping strategies, such as rumination or thought suppression, as key processes that maintain PTSD symptoms. Anxiety, depression and externalising symptoms can also present in children in the aftermath of trauma, yet there has been little empirical investigation of the potential relevance of posttrauma cognitive processes for such difficulties. Here, we examined whether: a) acute maladaptive cognitive processes (specifically, maladaptive appraisals, memory qualities, and cognitive coping) were associated with symptoms of PTSD, internalising, and externalising at 1-month posttrauma (T1); and b) changes in these cognitive processes predicted symptom change at a follow-up assessment 6 months later (T2). We recruited 132 6-13 year old children and their parents from emergency departments following the child's experience of an acute trauma. Children self-reported on their maladaptive appraisals, trauma-memory and cognitive coping strategies, along with symptoms of PTSD, anxiety and depression. Parents also rated children's internalising and externalising symptoms. We found each cognitive process to be robustly associated with PTSD and non-PTSD internalising symptoms at T1, and change in each predicted change in symptoms to T2. Maladaptive appraisals and cognitive coping were unique predictors of children's posttrauma internalising. Effects were partially retained even controlling for co-occurring PTSD symptoms. There was less evidence that trauma-specific cognitive processes were associated with externalising symptoms. Findings suggest aspects of cognitive models of PTSD are applicable to broader posttrauma psychopathology, and have implications for how we understand and target children's posttrauma psychological adjustment.


Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks.
In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV) concomitantly using two different enzyme immunoassay (EIA) tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of São Paulo (Fundação Pró-Sangue/Hemocentro de São Paulo), which is a major blood center in the city of São Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB) test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. Out of the 698 191 blood units that were donated, 2 718 of them (0.4%) had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58%) did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great majority of HIV-positive donors have already seroconverted and will be detected using only a single EIA screening test. Only persons who are going through the process of seroconversion may be missed by using a single EIA test and detected using two EIAs for screening. To screen out these individuals and decrease the residual risk of HIV transmission from the blood donated in our center, the use of other techniques, such as nucleic acid testing (NAT) or a p24 antigen assay, would be more effective.


Accuracy of Point-of-care Lung Ultrasonography for Diagnosis of Acute Chest Syndrome in Pediatric Patients with Sickle Cell Disease and Fever.
The objective was to determine the test performance characteristics for point-of-care lung ultrasonography (LUS) performed by pediatric emergency medicine (PEM) physicians compared with radiographic diagnosis of acute chest syndrome (ACS) in patients with sickle cell disease (SCD) and fever. This was a prospective study of patients up to 21 years with SCD and fever requiring chest X-ray (CXR) evaluation for ACS. Before obtaining CXR, a blinded PEM physician performed LUS using a standardized scanning protocol. Positive LUS for ACS was defined as lung consolidation. All patients received CXR and follow-up. The criterion standard for ACS was consolidation on CXR as determined by a blinded radiologist. LUS clips were reviewed by a blinded expert PEM sonologist. A total of 116 febrile events from 91 patients with a median age of 5.7 years were enrolled by 15 PEM sonologists. CXR was positive for ACS in 15 (13%) patients, and LUS was positive for ACS in 19 (16%) patients. Positive LUS had a sensitivity of 87% (95% confidence interval [CI] = 62% to 96%), specificity of 94% (95% CI = 88% to 97%), positive likelihood ratio of 14.6 (95% CI = 6.5 to 32.5), and negative likelihood ratio of 0.14 (95% CI = 0.04 to 0.52) for ACS. The interobserver agreement (kappa) was 0.77. There were two missed cases of ACS on LUS. LUS may be sensitive and specific for diagnosis of ACS in pediatric patients with SCD and fever. LUS may reduce the need for routine CXR and associated ionizing radiation exposure in this population.


Chemical and functional properties of metal chelators that mobilize copper to elicit fungal killing of Cryptococcus neoformans.
A panel of iron (Fe) and copper (Cu) chelators was screened for growth inhibitory activity against the fungal pathogen Cryptococcus neoformans. Select bidentate metal-binding ligands containing mixed O,S or O,N donor atoms were identified as agents that induce cell killing in a Cu-dependent manner. Conversely, structurally similar ligands with O,O donor atoms did not inhibit C. neoformans growth regardless of Cu status. Studies of Cu(ii) and Cu(i) binding affinity, lipophilicity, and growth recovery assays of Cu-import deficient cells identified lipophilicity of thermodynamically stable CuIIL2 complexes as the best predictor of antifungal activity. These same complexes induce cellular hyperaccumulation of Zn and Fe in addition to Cu. The results described here present the utility of appropriate metal-binding ligands as potential antifungal agents that manipulate cellular metal balance as an antimicrobial strategy.


Synthesis of cardiolipin derivatives with protection of the free hydroxyl: its application to the study of cardiolipin stimulation of cytochrome c oxidase.
Cardiolipin derivatives retaining the free hydroxyl on the polar head group were synthesized. With the use of a tetrahydropyranyl ether to protect this hydroxyl, fatty acyl substitutions were made at both of the 2-positions of cardiolipin (CL). The disubstituted derivatives were obtained in high yields. The stimulation of delipidated cytochrome c oxidase activity shows a hyperbolic dependence on the concentration of these CL derivatives. Both activation parameters, the apparent dissociation constant (Kd,app) and the maximum change in molecular activity (delta Actmax), depend on the chain length of the tails, with less dependence on the degree of saturation. Natural CL (92% C18:2, 8% C18:1) and CL disubstituted with oleic acid (47% C18:2, 52% C18:1) were equally effective at stimulating cytochrome c oxidase activity, with an apparent dissociation constant of approximately 1 microM when incubated in 0.3% Triton X-100 and assayed in lauryl maltoside. CL disubstituted with hexanoic acid, however, is a poorer activator, with an apparent dissociation constant of 6.8 microM and a delta Actmax that is 50% of that achieved with natural CL. Dilysocardiolipin, with complete removal of two of the fatty acid tails, shows negligible stimulation of cytochrome c oxidase activity.


Caries experience of children in primary schools with long-term tooth brushing programs: A pilot Australian study.
The aim of this study was to evaluate a primary school-based tooth brushing (TB) program conducted in a low socio-economic area of Queensland, Australia, to determine its effectiveness in reducing caries. Records kept at the central dental clinic of the district were used to analyse the caries experience (decayed, missing, filled teeth [dmft/DMFT]) and caries prevalence in children from two schools with long-term TB programs (TB) (N=1191) and three Non-TB schools (N=553). The schools were matched by socio-economic indices. Historical records showed that the baseline caries experience in all TB and Non-TB primary schools were similar at each primary school year. After a mean period of 5-9 years of the TB program, the caries experience (mean decayed, missing, filled teeth, dmft/DMFT) and prevalence were lower for TB group than Non-TB group. In the primary dentition, the overall mean dmft (±standard deviation) of TB group (2.53±3.00) was significantly lower than the Non-TB group (3.06±3.30) (P<.001). Similarly, in the permanent dentition, the overall mean DMFT of TB group (0.47±1.05) was reduced significantly compared to the Non-TB group (1.15±1.72) (P<.001). The overall caries prevalence in the TB group was 68% compared to 78% in Non-TB (P<.001). Overall, the mean annual DMFT increments of children in the TB schools were also significantly less compared with children in the Non-TB schools (P<.001). A long-term primary school TB program significantly reduced caries experience and caries prevalence in an optimally fluoridated (1-ppm), very low socio-economic district.


An exploration of physical activity and wellbeing in university employees.
The aim was to explore levels of physical activity (PA) and mental wellbeing in university employees, as well as barriers to and incentives for workplace PA. An electronic survey was distributed to all staff at one UK university. The survey consisted of a PA stages of change questionnaire, an international PA questionnaire (short-form), the Warwick-Edinburgh Mental Well-being Scale (WEMWBS), questions on perceived barriers to and incentives for workplace PA, questions on methods of enhancing employee wellbeing and demographics. A self-selected sample participated in two focus groups to explore key themes arising from the survey. Descriptive statistics were reported for survey data; associations between PA and wellbeing were tested for using Kruskal-Wallis with post hoc Mann-Whitney. Descriptive, thematic analysis was performed on focus group transcripts. A total of 502 surveys were completed (34% response rate); 13 staff participated in focus groups. In all, 42% of the sample reported PA below the recommended guideline amount. Females were less active than males (p < 0.005). The mean WEMWBS was 49.2 (95% confidence interval (CI): 48.3-49.9). Low PA levels were related to lower WEMWBS scores, with statistically significant differences in WEMWBS demonstrated between low and moderate PA (p = 0.05) and low and high PA (p = 0.001). Lack of time and perceived expense of facilities were common barriers to workplace PA. The main focus group finding was the impact of university culture on workplace PA and wellbeing. University staff demonstrate PA levels and a relationship between PA and wellbeing similar to the general population. Carefully designed strategies aimed at enhancing PA and wellbeing in university staff are required. The specific cultural and other barriers to workplace PA that exist in this setting should be considered. These results are being used to inform PA and wellbeing interventions whose effectiveness will be evaluated in future research.


Accurate predictions of individual differences in task-evoked brain activity from resting-state fMRI using a sparse ensemble learner.
Modelling and predicting individual differences in task-fMRI activity can have a wide range of applications from basic to clinical neuroscience. It has been shown that models based on resting-state activity can have high predictive accuracy. Here we propose several improvements to such models. Using a sparse ensemble learner, we show that (i) features extracted using Stochastic Probabilistic Functional Modes (sPROFUMO) outperform the previously proposed dual-regression approach, (ii) that the shape and overall intensity of individualised task activations can be modelled separately and explicitly, (iii) training the model on predicting residual differences in brain activity further boosts individualised predictions. These results hold for both surface-based analyses of the Human Connectome Project data as well as volumetric analyses of UK-biobank data. Overall, our model achieves state of the art prediction accuracy on par with the test-retest reliability of task-fMRI scans, suggesting that it has potential to supplement traditional task localisers.


Steroids in fluid and/or vasoactive infusion dependent pediatric shock: study protocol for a randomized controlled trial.
Physicians often administer corticosteroids for the treatment of fluid and vasoactive infusion dependent pediatric shock. This use of corticosteroids is controversial, however, and has never been studied in a pediatric randomized controlled trial (RCT). This pilot trial will determine the feasibility of a larger RCT on the role of corticosteroids in pediatric shock. Steroids in Fluid and/or Vasoactive Infusion Dependent Pediatric Shock (STRIPES) is a pragmatic, seven-center, double-blind, pilot RCT. We aim to randomize 72 pediatric patients with fluid and vasoactive infusion dependent shock to receive either hydrocortisone or a saline placebo for 7 days or until clinical stability, whichever occurs first. The primary outcome of this pilot trial is the feasibility of recruitment, defined as the number of patients enrolled over a 1-year period. Secondary outcomes include the frequency of, and reasons for, open-label steroid use, protocol adherence, incidence of mortality and corticosteroid-associated adverse events, time to discontinuation of inotropes, and feasibility of blood sampling. Corticosteroids are used for the treatment of pediatric shock without sufficient evidence to support this practice. While there is a scientific rationale and limited data supporting their use in this setting, there is also evidence from other populations suggesting potential harm. The STRIPES pilot study will assess the feasibility of a larger, much needed trial powered for clinically important outcomes. ClinicalTrials.gov: NCT02044159.


Extracellular matrix and androgen receptor expression associated with spontaneous transformation of rat prostate fibroblasts.
Spontaneous transformation in continuous culture of the androgen-sensitive rat prostate fibroblast cell line, NbF-1, resulted in an aggressively tumorigenic nonmetastatic phenotype that coincided with few gross chromosome abnormalities. This study identified transformation-associated alterations in extracellular matrix and androgen receptor expression in the NbF-1 cell line. Substantial levels of procollagens I, III, and IV and fibronectin mRNAs were detected in nontumorigenic NbF-1 cells. Laminin B1 and B2 mRNAs were also detectable, but at lower levels. Expression of all six extracellular matrix mRNAs was nonuniformly lower in tumorigenic NbF-1 cells. This decrease in expression was greatest for alpha 2 procollagen IV mRNA, which was reduced 17-fold. Proteoglycans and glycosaminoglycans synthesized by the NbF-1 cultures were also characterized. The NbF-1 cell line expressed chondroitin sulfate proteoglycans predominantly, and expression was reduced 5- to 10-fold in tumorigenic cultures. In contrast to the extensive alterations in the extracellular matrix, measurement of high-affinity androgen binding and androgen receptor mRNA levels showed substantial expression of androgen receptors in both NbF-1 cultures. Cultures of early and late passage NbF-1 cells demonstrated a mitogenic response to dihydrotestosterone. These data indicate (a) that alterations in expression of extracellular matrix components may represent early markers for tumorigenic transformation in prostatic mesenchymal cells and (b) that these changes can occur without disrupting androgen receptor expression and androgen sensitivity.


Combining structured and unstructured data to identify a cohort of ICU patients who received dialysis.
To develop a generalizable method for identifying patient cohorts from electronic health record (EHR) data-in this case, patients having dialysis-that uses simple information retrieval (IR) tools. We used the coded data and clinical notes from the 24,506 adult patients in the Multiparameter Intelligent Monitoring in Intensive Care database to identify patients who had dialysis. We used SQL queries to search the procedure, diagnosis, and coded nursing observations tables based on ICD-9 and local codes. We used a domain-specific search engine to find clinical notes containing terms related to dialysis. We manually validated the available records for a 10% random sample of patients who potentially had dialysis and a random sample of 200 patients who were not identified as having dialysis based on any of the sources. We identified 1844 patients that potentially had dialysis: 1481 from the three coded sources and 1624 from the clinical notes. Precision for identifying dialysis patients based on available data was estimated to be 78.4% (95% CI 71.9% to 84.2%) and recall was 100% (95% CI 86% to 100%). Combining structured EHR data with information from clinical notes using simple queries increases the utility of both types of data for cohort identification. Patients identified by more than one source are more likely to meet the inclusion criteria; however, including patients found in any of the sources increases recall. This method is attractive because it is available to researchers with access to EHR data and off-the-shelf IR tools.


Discovering multi-level structures in bio-molecular data through the Bernstein inequality.
The unsupervised discovery of structures (i.e. clusterings) underlying data is a central issue in several branches of bioinformatics. Methods based on the concept of stability have been recently proposed to assess the reliability of a clustering procedure and to estimate the "optimal" number of clusters in bio-molecular data. A major problem with stability-based methods is the detection of multi-level structures (e.g. hierarchical functional classes of genes), and the assessment of their statistical significance. In this context, a chi-square based statistical test of hypothesis has been proposed; however, to assure the correctness of this technique some assumptions about the distribution of the data are needed. To assess the statistical significance and to discover multi-level structures in bio-molecular data, a new method based on Bernstein's inequality is proposed. This approach makes no assumptions about the distribution of the data, thus assuring a reliable application to a large range of bioinformatics problems. Results with synthetic and DNA microarray data show the effectiveness of the proposed method. The Bernstein test, due to its loose assumptions, is more sensitive than the chi-square test to the detection of multiple structures simultaneously present in the data. Nevertheless it is less selective, that is subject to more false positives, but adding independence assumptions, a more selective variant of the Bernstein inequality-based test is also presented. The proposed methods can be applied to discover multiple structures and to assess their significance in different types of bio-molecular data.


Identifying research needs for improving health care.
Insights from systematic reviews can help new studies better meet the priorities and needs of patients and communities. However, systematic reviews unfortunately have not yet achieved this position to direct and guide new research studies. The Agency for Healthcare Research and Quality's Evidence-based Practice Center Program uses systematic reviews to identify gaps in current evidence and has developed a systematic process of prioritizing these gaps with stakeholder input into clearly defined "future research needs." Eight Evidence-based Practice Centers began to apply this effort in 2010 to various clinical and policy topics. Gaps that prevented systematic reviewers from answering central questions of the review may include insufficient studies on subpopulations, insufficient studies with appropriate comparators, lack of appropriate outcomes measured, and methods problems. Stakeholder panels, consisting of advocacy groups, patients, researchers, clinicians, funders, and policymakers, help refine the gaps through multiple conference calls and prioritization exercises. Each report highlights a focused set of 4 to 15 high-priority needs with an accompanying description of possible considerations for study design. Identification of high-priority research needs could potentially speed the development and implementation of high-priority, stakeholder-engaged research.


A prospective, non-randomized phase II trial of Trastuzumab and Capecitabine in patients with HER2 expressing metastasized pancreatic cancer.
Pancreatic cancer is the fourth most common cause of cancer related death in Western countries. Advantages in surgical techniques, radiation and chemotherapy had almost no impact on the long term survival of affected patients. Therefore, the need for better treatment strategies is urgent. HER2, a receptor tyrosine kinase of the EGFR family, involved in signal transduction pathways leading to cell growth and differentiation is overexpressed in a number of cancers, including breast and pancreatic cancer. While in breast cancer HER2 has already been successfully used as a treatment target, there are only limited data evaluating the effects of inhibiting HER2 tyrosine kinases in patients with pancreatic cancer. Here we report the design of a prospective, non-randomized multi-centered Phase II clinical study evaluating the effects of the Fluoropyrimidine-carbamate Capecitabine (Xeloda) and the monoclonal anti-HER2 antibody Trastuzumab (Herceptin) in patients with non-resectable, HER2 overexpressing pancreatic cancer. Patients eligible for the study will receive Trastuzumab infusions on day 1, 8 and 15 concomitant to the oral intake of Capecitabine from day 1 to day 14 of each three week cycle. Cycles will be repeated until tumor progression. A total of 37 patients will be enrolled with an interim analysis after 23 patients. Primary end point of the study is to determine the progression free survival after 12 weeks of bimodal treatment with the chemotherapeutic agent Capecitabine and the anti-HER2 antibody Trastuzumab. Secondary end points include patient's survival, toxicity analysis, quality of life, the correlation of HER2 overexpression and clinical response to Trastuzumab treatment and, finally, the correlation of CA19-9 plasma levels and progression free intervals.


Preclinical evaluation of the reinforcing and discriminative stimulus effects of agomelatine (S-20098), a melatonin agonist.
Agomelatine (S-20098), an analog of melatonin, has shown promise as a chronobiotic in animal models of sleep phase disorders and is being developed for clinical use. Previous research has shown that the pharmacological profile of melatonin-like drugs overlaps that of gamma-amino butyric acid (GABA) agonists. Given the potential of drugs within the latter class for recreational abuse in humans, evaluation of this potential for melatonin analogs that target similar therapeutic indications is important. In the present study, agomelatine was tested in animal models of the subjective and reinforcing effects of CNS depressant drugs; i.e., diazepam discrimination in rats and IV methohexital self-administration in rhesus monkeys, respectively. Neither agomelatine nor melatonin substituted for diazepam in rats trained to discriminate 2.5 mg/kg diazepam from vehicle. Further, agomelatine was not self-administered by rhesus monkeys. These results suggest that agomelatine would not produce diazepam-like intoxication in humans, nor would it likely be subject to abuse.


Planning and Evaluating Remote Consultation Services: A New Conceptual Framework Incorporating Complexity and Practical Ethics.
Establishing and running remote consultation services is challenging politically (interest groups may gain or lose), organizationally (remote consulting requires implementation work and new roles and workflows), economically (costs and benefits are unevenly distributed across the system), technically (excellent care needs dependable links and high-quality audio and images), relationally (interpersonal interactions are altered), and clinically (patients are unique, some examinations require contact, and clinicians have deeply-held habits, dispositions and norms). Many of these challenges have an under-examined ethical dimension. In this paper, we present a novel framework, Planning and Evaluating Remote Consultation Services (PERCS), built from a literature review and ongoing research. PERCS has 7 domains-the reason for consulting, the patient, the clinical relationship, the home and family, technologies, staff, the healthcare organization, and the wider system-and considers how these domains interact and evolve over time as a complex system. It focuses attention on the organization's digital maturity and digital inclusion efforts. We have found that both during and beyond the pandemic, policymakers envisaged an efficient, safe and accessible remote consultation service delivered through state-of-the art digital technologies and implemented via rational allocation criteria and quality standards. In contrast, our empirical data reveal that strategic decisions about establishing remote consultation services, allocation decisions for appointment type (phone, video, e-, face-to-face), and clinical decisions when consulting remotely are fraught with contradictions and tensions-for example, between demand management and patient choice-leading to both large- and small-scale ethical dilemmas for managers, support staff, and clinicians. These dilemmas cannot be resolved by standard operating procedures or algorithms. Rather, they must be managed by attending to here-and-now practicalities and emergent narratives, drawing on guiding principles applied with contextual judgement. We complement the PERCS framework with a set of principles for informing its application in practice, including education of professionals and patients.


Open chromatin in pluripotency and reprogramming.
Pluripotent stem cells can be derived from embryos or induced from adult cells by reprogramming. They are unique among stem cells in that they can give rise to all cell types of the body. Recent findings indicate that a particularly 'open' chromatin state contributes to maintenance of pluripotency. Two principles are emerging: specific factors maintain a globally open chromatin state that is accessible for transcriptional activation; and other chromatin regulators contribute locally to the silencing of lineage-specific genes until differentiation is triggered. These same principles may apply during reacquisition of an open chromatin state upon reprogramming to pluripotency, and during de-differentiation in cancer.


Formation of temporary flagellar structures during insect organogenesis.
Cilia and flagella are rare in nongerminal tissues of anthropods, and are generally thought to be restricted to sperm and sensory cells in insects (2). Whitten (5) has reported the presence of kinetosomes at the base of mitotrichia in the dipteran fly Sarcophaga bullata, but reports no evidence of the organization of fibrous elements characteristic of cilia and or flagella. During an ultrastructural analysis of morphogenesis of the colleterial gland of the silk moth Hyalophora cecropia, we found the first example of paired flagella associated with an insect secretory cell. These structures are also unusual in that they serve a temporary role in morphogenesis and subsequently disappear at the terminal stages of differentiation.


Design of photonic band gap fibers with suppressed higher-order modes: towards the development of effectively single mode large hollow-core fiber platforms.
The objective of the present investigation is to propose and theoretically demonstrate the effective suppression of higher-order modes in large-hollow-core photonic band gap fibers (PBGFs), mainly for low-loss data transmission platforms and/or high power delivery systems. The proposed design strategy is based on the index-matching mechanism of central air-core modes with defected outer core modes. By incorporating several air-cores in the cladding of the PBGF with 6-fold symmetry it is possible to resonantly couple the light corresponding to higher-order modes into the outer core, thus significantly increasing the leakage losses of the higher-order modes in comparison to the fundamental mode, thus making our proposed design to operate in an effectively single mode fashion with polarization independent propagation characteristics. The validation of the procedure is ensured with a detailed PBGF analysis based on an accurate finite element modal solver. Extensive numerical results show that the leakage losses of the higher-order modes can be enhanced in a level of at least 2 orders of magnitude in comparison to those of the fundamental mode. Our investigation is expected to remove an essential obstacle in the development of large-core single-mode hollow-core fibers, thus enabling them to surpass the attenuation of conventional fibers.


Characterization of betasatellite associated with the yellow mosaic disease of grain legumes in Southern India.
Yellow mosaic disease caused by mungbean yellow mosaic virus (MYMV) belonging to the genus Begomovirus (the family Geminiviridae) is a major constraint in cultivation of grain legumes in India. The urdbean (Vigna mungo (L.) Hepper) and mungbean (Vigna radiata (L.) R. Wilczek) samples affected with yellow mosaic disease exhibits yellow mosaic symptoms along with leaf puckering and leaf distortion in Tamil Nadu. Hence the study was performed to find out if there was any association and influence of betasatellite DNA on the symptom expression of MYMV. Full length viral clones of DNA A and DNA B were obtained through rolling circle amplification from YMD infected samples and identified as mungbean yellow mosaic virus. Interestingly, betasatellite was found to associate with MYMV, and its nucleotide sequence analysis showed its 95% identity with papaya leaf curl betasatellite (DQ118862) from cowpea. The present study represents the first report about the association of papaya leaf curl betasatellite with MYMV and represents a new member of the emerging group of bipartite begomovirus associated with betasatellite DNA.


Lifting the burden: a coordinated approach to action on Aboriginal tobacco resistance and control in NSW.
Smoking prevalence continues to be significantly higher among Aboriginal people than non-Aboriginal people, resulting in a range of serious health consequences and inequities. The Aboriginal Health and Medical Research Council of New South Wales (AHandMRC) and the New South Wales (NSW) Ministry of Health (the Ministry) have worked in partnership to develop The ATRAC Framework: A Strategic Framework for Aboriginal Tobacco Resistance and Control in NSW, in collaboration with Aboriginal communities and a range of stakeholders. The goal of the ATRAC Framework is to reduce smoking prevalence and the harmful impacts of tobacco use among Aboriginal people and communities in NSW. The framework includes reviews of relevant evidence and recommended actions, organised under six areas: leadership, partnerships and coordination; community action, awareness and engagement; workforce development; supportive environments; quitting support; and evidence, evaluation and research. The framework stresses that, to be successful, Aboriginal tobacco resistance and control programs and activities need to be evidence based, coordinated, integrated and involve Aboriginal people and Aboriginal community controlled health organisations in all aspects, from development through to implementation and evaluation. Consultations and evidence reviews highlight the importance of workforce support and development, including the ongoing need for more workers specialising in Aboriginal tobacco resistance and control, as well as ongoing training for all staff involved in delivering care to Aboriginal people. Other key strategies identified in the framework include improving access to nicotine replacement therapy and other medications to support quitting; supporting, strengthening and building on existing innovative community-based programs; and further developing the evidence base. The AHandMRC and the Ministry will continue to work in partnership to drive the use of the ATRAC Framework by all people involved in Aboriginal tobacco resistance and control in NSW for resource allocation, planning, implementation and evaluation of existing and future activities.


The effect of biomechanical properties of motion on infants' perception of goal-directed grasping actions.
From a very young age, infants perceive others' actions as goal directed. Yet, the processes underlying this competence are still debated. In this study, we investigated whether (a) 4- and 6-month-old infants and adults discriminate the biomechanical properties of the human hand within an action context, (b) the manipulation of the biomechanics of hand movements has an impact on the ability to anticipate the goal of an action, and (c) the emergence of motor experience with grasping is related to infants' ability to discriminate the biomechanics of hand movements and to anticipate the action goal. The 6-month-olds discriminated between biomechanically possible and impossible grasps, and in some (but not all) instances they made more anticipatory gaze shifts toward the goal of the possible action. Both the 4- and 6-month-olds' processing of biomechanical properties of the hand were significantly related to their ability to anticipate the goal of a grasping action. Importantly, those 4-month-olds with higher precision grasping skills manifested faster anticipatory gazes toward the goal of the action. These findings suggest that multiple sources of information from an action scene are interdependent and that both perceptual information and motor experience with an action are relevant for on-line prediction of the final goal of the action.


Epidemiologic trend in elderly domestic injury.
The elderly constitute about 12% of the American population, with a projected increase of up to 25% in 2050. Elderly domestic injuries have been recognized as a major cause of morbidity and mortality. The objective of this study is to determine the 4-y national trend in elderly domestic injury, and we hypothesize that the home remains a significant source of injury. Data on elderly patients ≥ 65 y was extracted from the National Trauma Data Bank's National Sample Project (NSP). Elderly patients with home injuries were compared with non-home injuries. Subsets of hospitalized patients were analyzed for trends in injury site over a 4 y period. Multivariate analysis was performed to determine the predictors of hospitalization and in-hospital mortality. A total of 98,288 patients, representing a weighed estimate of 472,456 elderly patients were analyzed. Forty-two percent of all injuries in the study population occurred at home, followed by motor vehicle crashes (MVC) at 25%. Home injuries as a proportion of annual injuries increased from 37% in 2003 to 40% in 2006. Majority (57%) were admitted to the floor and 14% to the intensive care unit (ICU). On multivariate analysis, African-Americans and Asians were less likely to be hospitalized (odds ratio (OR) 0.57 and 0.50, respectively, with females 47% less likely than males to die after hospitalization (P < 0.001). Home injuries remain the most significant source of elderly hospitalizations after trauma. With a rapidly growing elderly population, there is a need to recognize this specific location of injury and create directed preventive measures to avert elderly domestic injuries.


Redetermination of katayamalite, KLi3Ca7Ti2(SiO3)12(OH)2.
The crystal structure of katayamalite, ideally KLi3Ca7Ti2(SiO3)12(OH)2 (potassium trilithium hepta-calcium dititanium dodeca-silicate di-hydroxide), was previously reported in triclinic symmetry (C-1), with isotropic displacement parameters for all atoms and without the H-atom position [Kato & Murakami (1985 ▶). Mineral. J. 12, 206-217]. The present study redetermines the katayamalite structure with monoclinic symmetry (space group C2/c) based on single-crystal X-ray diffraction data from a sample from the type locality, Iwagi Island, Ehime Prefecture, Japan, with anisotropic displacement parameters for all non-H atoms, and with the H atoms located by difference Fourier analysis. The structure of katayamalite contains a set of six-membered silicate rings inter-connected by sheets of Ca atoms on one side and by an ordered mixture of Li, Ti and K atoms on the other side, forming layers which are stacked normal to (001). From the eight different metal sites, three are located on special positions, viz. one K and one Li atom on twofold rotation axes and one Ca atom on an inversion center. The Raman spectrum of kataymalite shows a band at 3678 cm(-1), similar to that observed for hydroxyl-amphiboles, indicating no or very weak hydrogen bonding.


Selection of xenobiotic-degrading microorganisms in a biphasic aqueous-organic system.
Microbial selection on mixtures of chlorinated and nonchlorinated compounds that are poorly soluble in water and/or toxic to growing microbial cells was examined in both biphasic aqueous-organic and monophasic aqueous systems. A biphasic system in which silicone oil was used as the organic phase permitted the acceleration of acclimation, leading to rapid selection and to an increase in xenobiotic compound degradation. In contrast, acclimation, selection, and degradation were very slow in the monophasic aqueous system. The variation in microbial growth rate with the degree of dispersion (i.e., dispersion at different silicone oil concentrations and agitation rates), and cell adhesion to the silicone oil indicate that the performance of the biphasic aqueous-organic system is dependent on the interfacial area between the two phases and that microbial activity is important at this interface. Therefore, the biphasic water-silicone oil system could be used for microbial selection in the presence of xenobiotic compounds that are toxic and have low water solubility.


GAIP interacting protein C-terminus regulates autophagy and exosome biogenesis of pancreatic cancer through metabolic pathways.
GAIP interacting protein C terminus (GIPC) is known to play an important role in a variety of physiological and disease states. In the present study, we have identified a novel role for GIPC as a master regulator of autophagy and the exocytotic pathways in cancer. We show that depletion of GIPC-induced autophagy in pancreatic cancer cells, as evident from the upregulation of the autophagy marker LC3II. We further report that GIPC regulates cellular trafficking pathways by modulating the secretion, biogenesis, and molecular composition of exosomes. We also identified the involvement of GIPC on metabolic stress pathways regulating autophagy and microvesicular shedding, and observed that GIPC status determines the loading of cellular cargo in the exosome. Furthermore, we have shown the overexpression of the drug resistance gene ABCG2 in exosomes from GIPC-depleted pancreatic cancer cells. We also demonstrated that depletion of GIPC from cancer cells sensitized them to gemcitabine treatment, an avenue that can be explored as a potential therapeutic strategy to overcome drug resistance in cancer.


Innate Immunity of Neonates and Infants.
Many important events occur at birth. The fetus is suddenly removed from a protected intra-uterine environment that is aquatic, warm, and nearly sterile, to the dry, cold external world laden with microbes. To survive, the neonate must interact with many organisms, making use of some, while vigorously defending against the others like a nation conducting trade with friendly countries and guarding against hostile ones from invading it, waging wars if necessary. Although, the neonatal immune system is plastic, however, it is highly tolerant which is due to both the fetal development during gestation as well as significant sudden changes in fetal environment and enormous exposure to the new antigens and intestinal bacteria and their products. This "quiescent mode" of innate immune system is part of a highly regulated process to fulfill all requirements of multi-layered process of early life, implemented effectively through the cells of innate immune system. While, most of the neonatal innate immune cells (e.g., neutrophils and monocytes) present contained activity and lower frequencies compared to their adult counterparts, innate lymphoid cells (ILCs), a distinct cellular component of innate immunity, show higher level of activity and presence during period of infancy compared to later stages of life and adulthood, which may suggest a role for ILCs in variable susceptibility to certain conditions during life time. In this review, while we focus on the characteristics and status of ILCs in neonatal immune system, we also draw an analogy from a national defense perspective because of the great similarities between that and the immune system by providing the known biological counterparts of all five core operational elements, the five Ds of defense, detection, discrimination, deployment, destruction, and de-escalation, with special focus on innate immunity, maternal support, and influence during the neonatal and infancy periods.


Controlling electron transfer through the manipulation of structure and ligand-based torsional motions: a computational exploration of ruthenium donor-acceptor systems using density functional theory.
Computational studies using density functional theory (DFT) are reported for a series of donor-acceptor (DA) transition metal complexes and related excited-state and electron transfer (ET) photoproduct models. Three hybrid Hartree-Fock/DFT (HF/DFT) functionals, B3LYP, B3PW91, and PBE1PBE, are employed to characterize structural features implicated in the dynamical control of productive forward and energy wasting back ET events. Energies and optimized geometries are reported for the lowest energy singlet state in [Ru(dmb)(2)(bpy-phi-MV)](4+) (DA1), [Ru(dmb)(2)(bpy-o-tolyl-MV)](4+) (DA2), [Ru(dmb)(2)(bpy-2,6-Me(2)-phi-MV)](4+) (DA3), and [Ru(tmb)(2)(bpy-2,6-Me(2)-phi-MV)](4+) (DA3'), where dmb is 4,4'-dimethyl-2,2'-bipyridine, tmb is 4,4',5,5'-tetramethyl-2,2'-bipyridine, MV is methyl viologen, and phi is a phenylene spacer. These indicate that the dihedral angle theta(1) between the aryl substituent and the bipyridine fragment to which it is bound, systematically increases with the addition of steric bulk. Energies, optimized geometries, and unpaired electron spin densities are also reported for the lowest energy triplet state of [Ru(dmb)(2)(4-p-tolyl-2,2'-bipyridine)](2+) (D1*), [Ru(dmb)(2)(4-(2,6-dimethylphenyl)-2,2'-bipyridine)](2+) (D2*), [Ru(dmb)(2)(4-mesityl-2,2'-bipyridine)](2+) (D3*), and [Ru(tmb)(2)(4-mesityl-2,2'-bipyridine)](2+) (D3'*). Each of these serves as a model of a reactant excited state in the forward electron-transfer photochemistry allowing us to qualify and quantify the role of excited-state intraligand electron delocalization in driving substantial geometry changes (especially with respect to theta(1)) relative to its respective DA counterpart. Next, energies, optimized geometries, and spin densities are reported for the lowest energy triplet of each DA species: (3)DA1, (3)DA2, (3)DA3, and (3)DA3'. These are used to model the ET photoproduct and they indicate that theta(1) increases following ET, thus, verifying switch-like properties. Finally, we report data for geometry optimized DA1 and (3)DA1 in a continuum model of room temperature acetonitrile. This study shows a complete recovery of theta(1) to its ground state value which has implications in efforts to trap electrons in charge-separated states.


Novel diffusion tractography methodology using Kalman filter prediction to improve preoperative benefit-risk analysis in pediatric epilepsy surgery.
In this study the authors investigated the clinical reliability of diffusion weighted imaging maximum a posteriori probability (DWI-MAP) analysis with Kalman filter prediction in pediatric epilepsy surgery. This approach can yield a suggested resection margin as a dynamic variable based on preoperative DWI-MAP pathways. The authors sought to determine how well the suggested margin would have maximized occurrence of postoperative seizure freedom (benefit) and minimized occurrence of postoperative neurological deficits (risk). The study included 77 pediatric patients with drug-resistant focal epilepsy (age 10.0 ± 4.9 years) who underwent resection of their presumed epileptogenic zone. In preoperative DWI tractography from the resected hemisphere, 9 axonal pathways, Ci=1-9, were identified using DWI-MAP as follows: C1-3 supporting face, hand, and leg motor areas; C4 connecting Broca's and Wernicke's areas; C5-8 connecting Broca's, Wernicke's, parietal, and premotor areas; and C9 connecting the occipital lobe and lateral geniculate nucleus. For each Ci, the resection margin, di, was measured by the minimal Euclidean distance between the voxels of Ci and the resection boundary determined by spatially coregistered postoperative MRI. If Ci was resected, di was assumed to be negative (calculated as -1 × average Euclidean distance between every voxel inside the resected Ci volume, ri). Kalman filter prediction was then used to estimate an optimal resection margin, d*i, to balance benefit and risk by approximating the relationship between di and ri. Finally, the authors defined the preservation zone of Ci that can balance the probability of benefit and risk by expanding the cortical area of Ci up to d*i on the 3D cortical surface. In the whole group (n = 77), nonresection of the preoperative preservation zone (i.e., actual resection margin d*i greater than the Kalman filter-defined d*i) accurately predicted the absence of postoperative motor (d*1-3: 0.93 at seizure-free probability of 0.80), language (d*4-8: 0.91 at seizure-free probability of 0.81), and visual deficits (d*9: 0.90 at seizure-free probability of 0.75), suggesting that the preservation of preoperative Ci within d*i supports a balance between postoperative functional deficit and seizure freedom. The subsequent subgroup analyses found that preservation of preoperative Ci =1-4,9 within d*i =1-4,9 may provide accurate deficit predictions independent of age and seizure frequency, suggesting that the DWI-based surgical margin can be effective for surgical planning even in young children and across a range of epilepsy severity. Integrating DWI-MAP analysis with Kalman filter prediction may help guide epilepsy surgery by visualizing the margins of the eloquent white matter pathways to be preserved.


Simple, reliable, and nondestructive method for the measurement of vacuum pressure without specialized equipment.
We present a simple, reliable, and nondestructive method for the measurement of vacuum pressure in a magneto-optical trap. The vacuum pressure is verified to be proportional to the collision rate constant between cold atoms and the background gas with a coefficient k, which can be calculated by means of the simple ideal gas law. The rate constant for loss due to collisions with all background gases can be derived from the total collision loss rate by a series of loading curves of cold atoms under different trapping laser intensities. The presented method is also applicable for other cold atomic systems and meets the miniaturization requirement of commercial applications.


Ankyrin repeat and SOCS box containing protein 4 (Asb-4) colocalizes with insulin receptor substrate 4 (IRS4) in the hypothalamic neurons and mediates IRS4 degradation.
The arcuate nucleus of the hypothalamus regulates food intake. Ankyrin repeat and SOCS box containing protein 4 (Asb-4) is expressed in neuropeptide Y and proopiomelanocortin (POMC) neurons in the arcuate nucleus, target neurons in the regulation of food intake and metabolism by insulin and leptin. However, the target protein(s) of Asb-4 in these neurons remains unknown. Insulin receptor substrate 4 (IRS4) is an adaptor molecule involved in the signal transduction by both insulin and leptin. In the present study we examined the colocalization and interaction of Asb-4 with IRS4 and the involvement of Asb-4 in insulin signaling. In situ hybridization showed that the expression pattern of Asb-4 was consistent with that of IRS4 in the rat brain. Double in situ hybridization showed that IRS4 colocalized with Asb-4, and both Asb-4 and IRS4 mRNA were expressed in proopiomelanocortin (POMC) and neuropeptide Y (NPY) neurons within the arcuate nucleus of the hypothalamus. In HEK293 cells co-transfected with Myc-tagged Asb-4 and Flag-tagged IRS4, Asb-4 co-immunoprecipitated with IRS4; In these cells endogenous IRS4 also co-immunoprecipitated with transfected Myc-Asb-4; Furthermore, Asb-4 co-immunoprecipitated with IRS4 in rat hypothalamic extracts. In HEK293 cells over expression of Asb-4 decreased IRS4 protein levels and deletion of the SOCS box abolished this effect. Asb-4 increased the ubiquitination of IRS4; Deletion of SOCS box abolished this effect. Expression of Asb-4 decreased both basal and insulin-stimulated phosphorylation of AKT at Thr308. These data demonstrated that Asb-4 co-localizes and interacts with IRS4 in hypothalamic neurons. The interaction of Asb-4 with IRS4 in cell lines mediates the degradation of IRS4 and decreases insulin signaling.


Intracellular pH regulates bovine sperm motility and protein phosphorylation.
Bovine sperm in neat caudal epididymal fluid become motile in response to either pH elevation or dilution of the fluid. Buffers containing permeant weak acids at physiologic concentrations are able to mimic these effects of caudal fluid. These observations lead to the hypothesis that a pH-dependent epididymal fluid quiescence factor regulates bovine sperm motility by modulating sperm intracellular pH (pHi). Here we report that sperm pHi, measured with the fluorescent pH probe carboxyfluorescein, increases by approximately 0.4 units in response to either of these motility-initiating manipulations. At least 26 discrete phosphoprotein bands are distinguishable by sodium dodecylsulfate-polyacrylamide gel electrophoresis after incubation of intact caudal sperm with 32PO4. A prominent phosphoprotein, with Mr approximately 255,000 (pp255) and a relatively high specific radioactivity, is reversibly dephosphorylated in response to elevations in pHi that initiate sperm motility. Unlike most of the sperm phosphoproteins, the extraction of pp255 requires reducing agents. This phosphoprotein cosediments with the sperm heads but not the tail, midpiece, soluble, or plasma membrane fractions. No other pHi-dependent phosphorylation changes are apparent in gels of whole sperm extracts. However, subcellular fractionation allows the detection of increased phosphorylation of two plasma membrane phosphoproteins (Mr approximately 105,000 and 97,000) and decreased phosphorylation of another plasma membrane phosphoprotein (Mr approximately 120,000) in response to increasing pHi. This is the first report describing changes in endogenous phosphoproteins from intact motile and nonmotile bovine sperm that are regulated by pHi.


Adherence to antiretroviral medications among persons who inject drugs in transitional, low and middle income countries: an international systematic review.
Adherence to antiretroviral (ART) medication is vital to reducing morbidity and mortality among HIV positive persons. People who inject drugs (PWID) are at high risk for HIV infection in transitional/low/middle income countries (TLMIC). We conducted a systematic review of studies reporting adherence to ART among persons with active injection drug use and/or histories of injection drug use in TLMIC. Meta-regression was performed to examine relationships between location, adherence measurements, and follow-up period. Fifteen studies were included from seven countries. Adherence levels ranged from 33 to 97 %; mean weighted adherence was 72 %. ART adherence was associated with different methods of measuring adherence and studies conducted in Eastern Europe and East Asia. The great heterogeneity observed precludes generalization to TLMIC as a whole. Given the critical importance of ART adherence more research is needed on ART adherence among PWID in TLMIC, including the use of standardized methods for reporting adherence to ART.


Cartilage reinforcement tympanoplasty: otological and audiological results.
High risk perforations including total, subtotal, atelectatic perforations, those with cholesteatoma and revision cases can be treated efficiently with cartilage reinforcement tympanoplasty. The purpose of this study was to describe the perforation closure rates and hearing results of cartilage reinforcement tympanoplasty. This was a retrospective chart review and included 60 patients who underwent surgery with the cartilage reinforcement technique between November 2006 and October 2008 at the Department of Otorhinolaryngology of Haydarpasa Numune Research and Training Hospital, Istanbul, Turkey. The male to female ratio was 28 (46.7%)/32 (53.3%). The mean age of the patients was 30.10 +/- 11.50 years (range 13-55). Preoperatively, only one patient had a retraction pocket but not a perforation, 12 (20%) patients had perforation of < 25% of the total tympanic membrane diameter, 12 patients had perforation between 25% and 50% of the total membrane diameter, 18 patients had a perforation between 50% and 75% of total membrane diameter and 17 patients had a perforation of > 75% of the total membrane diameter. Postoperatively we achieved total closure of tympanic membrane perforations for 47 patients but the perforations of 13 patients remained in various sizes (7 patients had tympanic membrane perforation < 25% of the total membrane diameter, 3 between 25% and 50%, 2 between 50% and 75% and 1 patient had a perforation > 75% of the total membrane diameter). There was a statistically significant difference between percentages of preoperative perforation and postoperative perforations (p = 0.001). Only 1.7% of patients had no perforation preoperatively and this ratio increased to 78.3% postoperatively. Also, 30% of patients had a perforation between 50% and 75% of the total membrane diameter, 28.3% of the patients had a perforation > 75% of the total membrane diameter preoperatively and this ratio decreased to 3.3% and 1.7%, respectively. Concerning the audiological parameters, the difference between preoperative and postoperative hearing levels was statistically significant (p = 0.001).


High urinary corticoid/creatinine ratios in cats with hyperthyroidism.
Measurement of the urinary corticoid : creatinine (C : C) ratio provides an assessment of cortisol secretion over a period of time. Therefore, this test is a very sensitive measure of adrenocortical function. The stress of the diagnostic procedure and nonadrenal disease may increase the urinary C : C ratio. In addition, diseases such as hyperthyroidism may influence the metabolic clearance of cortisol. To evaluate the effect of thyroid hormone excess, urinary C : C ratios were measured in 32 cats with hyperthyroidism and 45 healthy household cats. In 7 cats, urinary C : C ratios were measured both before and after treatment for hyperthyroidism. With data from the healthy cats, the reference range for the urinary C : C ratio was determined to be 8.0 to 42.0 X 10(-6). The urinary C : C ratios in the cats with hyperthyroidism (median, 37.5 x 10(-6); range, 5.9-169.5 x 10(-6)) were significantly (P = .001) higher than those in the healthy cats (median, 16 x 10(-6); range, 4.8-52.5 x 10(-6)). In 15 cats with hyperthyroidism, the urinary C : C ratios exceeded the upper limit of the reference range. Treatment for hyperthyroidism led to a marked decrease in urinary C : C ratios. The results of this study demonstrate that the urinary C : C ratio may be abnormally high in cats with hyperthyroidism, probably because of increased metabolic clearance of cortisol and activation of the pituitary-adrenocortical axis by disease. Although the clinical features of hyperthyroidism and hyperadrenocorticism in cats are different, hyperthyroidism should be ruled out when cats are suspected of hyperadrenocorticism on the basis of abnormally high urinary C : C ratios.


A densely sampled ITS phylogeny of the Cape flagship genus Erica L. suggests numerous shifts in floral macro-morphology.
Erica L. is the largest of the 'Cape' clades that together comprise around half of the disproportionately high species richness of the Cape Floristic Region (CFR) of South Africa. Around 840 species of Erica are currently recognised, C.680 of which are found in the CFR, the rest distributed across the rest of Southern Africa, the highlands of Tropical Africa and Madagascar, and Europe. Erica is taxonomically well documented, but very little is known about species-level relationships. We present the first densely sampled phylogenetic analysis of Erica, using nuclear ribosomal DNA sequences (internal transcribed spacers; ITS) of c. 45% of the species from across the full geographic range of the genus, both Calluna and Daboecia (Ericeae; monotypic genera and putative sister groups of Erica), and further Ericoideae outgroups. Our results show both morphological and geographic coherence of some clades, but numerous shifts in floral macro-morphology as represented by variation in individual morphological characters and pollination syndromes. European Ericeae is a paraphyletic grade subtending a monophyletic African/Malagasy Erica. Given the limited resolution of this single gene tree, more data are needed for further conclusions. Clades are identified that will serve as an effective guide for targeted sampling from multiple linkage groups.


Mechanisms of the adenosine A2A receptor-induced sensitization of esophageal C fibers.
Clinical studies indicate that adenosine contributes to esophageal mechanical hypersensitivity in some patients with pain originating in the esophagus. We have previously reported that the esophageal vagal nodose C fibers express the adenosine A2A receptor. Here we addressed the hypothesis that stimulation of the adenosine A2A receptor induces mechanical sensitization of esophageal C fibers by a mechanism involving transient receptor potential A1 (TRPA1). Extracellular single fiber recordings of activity originating in C-fiber terminals were made in the ex vivo vagally innervated guinea pig esophagus. The adenosine A2A receptor-selective agonist CGS21680 induced robust, reversible sensitization of the response to esophageal distention (10-60 mmHg) in a concentration-dependent fashion (1-100 nM). At the half-maximally effective concentration (EC50: ≈3 nM), CGS21680 induced an approximately twofold increase in the mechanical response without causing an overt activation. This sensitization was abolished by the selective A2A antagonist SCH58261. The adenylyl cyclase activator forskolin mimicked while the nonselective protein kinase inhibitor H89 inhibited mechanical sensitization by CGS21680. CGS21680 did not enhance the response to the purinergic P2X receptor agonist α,β-methylene-ATP, indicating that CGS21680 does not nonspecifically sensitize to all stimuli. Mechanical sensitization by CGS21680 was abolished by pretreatment with two structurally different TRPA1 antagonists AP18 and HC030031. Single cell RT-PCR and whole cell patch-clamp studies in isolated esophagus-specific nodose neurons revealed the expression of TRPA1 in A2A-positive C-fiber neurons and demonstrated that CGS21682 potentiated TRPA1 currents evoked by allylisothiocyanate. We conclude that stimulation of the adenosine A2A receptor induces mechanical sensitization of nodose C fibers by a mechanism sensitive to TRPA1 antagonists indicating the involvement of TRPA1.


Applying novel methods to assess clinical outcomes: insights from the TRILOGY ACS trial.
Several methods provide new insights into understanding clinical trial composite endpoints, using both conventional and novel methods. The TRILOGY ACS trial is used as a contemporary example to prospectively compare these methods side by side. The traditional time-to-first-event, Andersen-Gill recurrent events method, win ratio, and a weighted composite endpoint (WCE) are compared using the randomized, active-control TRILOGY ACS trial. This trial had a neutral result and randomized 9326 patients managed without coronary revascularization within 10 days of their acute coronary syndrome to receive either prasugrel or clopidogrel and followed them for up to 30 months. The traditional composite, win ratio, and WCE demonstrated no significant survival advantage for prasugrel, whereas the Andersen-Gill method demonstrated a statistical advantage for prasugrel [hazard ratio (HR), 0.86 (95% CI, 0.72-0.97)]. The traditional composite used 73% of total patient events; 40% of these were derived from the death events. The win ratio used 66% of total events; deaths comprised 57% of these. Both Andersen-Gill and WCE methods used all events in all participants; however, with the Andersen-Gill method, death comprised 41% of the proportion of events, whereas with the WCE method, death comprised 64% of events. This study addresses the relative efficiency of various methods for assessing clinical trial events comprising the composite endpoint. The methods accounting for all events, in particular those incorporating their clinical relevance, appear most advantageous, and may be useful in interpreting future trials. This clinical and statistical advantage is especially evident with long-term follow-up where multiple non-fatal events are more common. NCT00699998.


Light-mediated antifungal activity of Echinacea extracts.
This study demonstrated that plant extracts containing acetylenic isobutylamides and polyacetylenes, previously reported as occurring in Echinacea, have phototoxic antimicrobial activity against fungi, including clinically relevant pathogenic fungi. Results show that hexane extracts of Echinacea variably inhibit growth of yeast strains of Saccharomyces cerevisiae, Candida shehata, C. kefyr, C. albicans, C. steatulytica and C. tropicalis under near UV irradiation (phototoxicity) and to a lower extent without irradiation (conventional antifungal activity). The presence of polyacetylenes and alkylamides in extracts of different organs was confirmed in Echinacea purpurea by HPLC in agreement with previously reported data in the literature, and was related to phototoxic activity. Two representative pure compounds, undeca-2E,4Z-diene-8,10-diynoic acid isobutylamide and dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, were isolated from Echinacea purpurea root extracts, and compared in a disk assay (5 micrograms/disk) with the highly conjugated trideca-1-ene-3,5,7,9,10-pentayne (previously isolated in our laboratory and found here in E. purpurea). Significant phototoxicity was demonstrated by pure trideca-1-ene-3,5,7,9,10-pentayne, while only minor phototoxicity was induced by the other two acetylenic compounds. Phototoxic activity of Echinacea spp. is primarily attributed to the ketoalkenes and ketoalkynes abundantly present in the roots.


Directed differentiation of umbilical cord blood stem cells into cortical GABAergic neurons.
Umbilical cord blood contains a population of non-hematopoietic multipotent stem cells that are capable of neuronal differentiation in-vitro. These cells have shown great potential as a therapeutic tool for central nervous system diseases and disorders. However whether these cells are able to produce neurons with similar developmental and functional characteristics to indigenous neurons within the brain remains poorly investigated. In this study, we used purified umbilical cord blood non-hematopoietic stem cells to produced GABAergic neurons with similar developmental and functional characteristics to cortical GABAergic neurons. We analyzed the expression of transcription factors MASH1, DLX1 and DLX2 throughout the 24 days of a sequential neuronal induction protocol and found that their expression patterns resembled those reported in the developing human cortex. The derived neurons also expressed components of GABAergic neurotransmission including GABA regulatory enzymes, GABA receptor subunits and GABA transporters. Thus we have demonstrated that umbilical cord blood stem cells are capable of producing cortical-like GABAergic neurons in vitro. This highlights the potential of umbilical cord blood stem cells as a therapeutic tool for neural injuries and disorders.