Depressed patients' perspectives of 2 measures of outcome: the Quick Inventory of Depressive Symptomatology (QIDS) and the Remission from Depression Questionnaire (RDQ).
Current operational definitions of remission, at their root, are exclusively symptom-based and therefore limited in scope. In this report from the Rhode Island Methods to Improve Diagnostic Assessment and Services (MIDAS) project we examined patient acceptability of a new measure, the Remission from Depression Questionnaire (RDQ), a scale designed to capture a broader array of domains patients consider relevant to the construct of remission. Patient acceptance of the RDQ was compared with that of the Quick Inventory of Depressive Symptomatology (QIDS), the instrument used to measure outcome in the Sequenced Treatment Alternatives to Relieve Depression study. One hundred and two depressed outpatients in ongoing treatment completed the RDQ, QIDS, and a 9-item measure of patient preference. Patients indicated the RDQ was a better indicator of their overall state and treatment goals. Patients judged the RDQ to be a more accurate and preferred measure to determine treatment outcome, and a more accurate indicator of remission. Patients considered the multifactorial RDQ a more accurate indicator of their treatment goals than a purely symptom measure such as the QIDS.


Dietary diversity and fish consumption of mothers and their children in fisher households in Komodo District, eastern Indonesia.
Small-scale coastal fisheries contribute directly and indirectly to the food and nutrition security of marine-dependent households. Fishers can apportion part of their catch for household consumption or use the income earned to purchase staples and other desired foods. Fish are an important animal-source food rich in micronutrients essential for cognitive development of children and for adult health, and a valuable addition to rice-based diets. Furthermore, the engagement of women in fisheries value chains and increased control over income may facilitate decision-making which improves nutrition outcomes for women and their children. Despite these contributions, food insecurity remains prevalent in many low and middle income fish-producing countries. This paper reports findings from an exploration of the interplaying factors leading to food and nutrition insecurity in three marine-dependent coastal communities in eastern Indonesia, focusing on the consumption pathway, that is, the contribution of fish to the diets and nutrition of women and children. The research was undertaken as a mixed-methods case study. The study found that over 50% of mother-child pairs failed to meet the minimum recommended dietary diversity, and, while fish was the main animal-source food in diets, the introduction of fish to infant and young child diets was delayed due to fears of allergies and illnesses. Moreover, access to nutrient-dense foods was affected by variable and insufficient income from fisheries-based livelihoods, isolation from markets, and the broader food environment. Given the shift towards 'nutrition-sensitive interventions' to improve the livelihoods and well-being of fisher households, these results highlight the need for analysis of the intra-household sharing of fish within fisher households, culturally-appropriate strategies to improve the quality of family and especially complementary foods, and efforts to increase physical access to nutrient-dense foods.


Assessment of the performance of the Stanford Online Calculator for the prediction of nonsentinel lymph node metastasis in sentinel lymph node-positive breast cancer patients.
Several models for the prediction of nonsentinel lymph node (NSLN) metastasis in sentinel lymph node (SLN)-positive breast cancer patients have been proposed. In this study, the authors evaluate the Stanford Online Calculator (SOC), which was designed to predict the likelihood of NSLN metastasis using only 3 variables: primary tumor size, SLN metastasis size, and angiolymphatic invasion status. They compared it with the Mayo and Memorial Sloan-Kettering Cancer Center (MSKCC) nomograms. The SOC was used to calculate the probability of NSLN metastasis in 464 breast cancer patients with SLN metastasis who underwent completion axillary lymph node dissection at the Mayo Clinic. The area under the receiver operating characteristic curve (AUC) was calculated for each model. Mean probabilities of patients with and without NSLN metastasis were compared. Patients with <or=5%, <or=10%, and 100% NSLN metastasis probabilities were examined. The AUCs of the Stanford, MSKCC, and Mayo models were 0.72, 0.74, and 0.77, respectively (P=.13). The mean Stanford probabilities for patients with and without NSLN metastasis were 0.75 (range, 0.06-1.0) and 0.50 (range, 0.05-1.0), respectively (P<.0001). The false-negative rates for patients with a Stanford probability of <or=5% and <or=10% were 0% and 13%, respectively. Of the patients with a Stanford probability of 100%, 26% did not have NSLN metastasis. Despite using only 3 variables, the Stanford nomogram appears to perform on a par with, but not better than, the MSKCC and Mayo nomograms. Further validation in other patient populations is needed.


Serum opsonic activity in rodent malaria: functional and immunochemical characteristics in vitro.
The functional and immunochemical characteristics of serum opsonic activity in rodent malaria were examined in the present study. Schizont- and late trophozoite-enriched populations of Plasmodium berghei-infected red blood cells (IRBC) were isolated on a Ficoll density-gradient and used in an in vitro phagocytosis system composed of serum and monolayer cultures of rat peritoneal macrophages. Hyperimmune serum augmented the phagocytosis of IRBC to a greater degree than did nonimmune serum. When either IRBC or macrophages were pre-incubated with serum, the phagocytosis-promoting factors acted on the IRBC rather than on the macrophages in a manner characteristic of serum opsonins. The opsonic activity was specific for IRBC since noninfected red blood cells were rarely phagocytized and were unable to absorb opsonic activity from serum. The opsonic activity of both hyperimmune and nonimmune sera was heat stable, and unaffected by agents known to inactivate or inhibit complement (cobra venom factor and ethylenediaminetetraacetic acid). Finally, the opsonic activity was identified in preparations of purified IgG isolated from both hyperimmune and nonimmune sera.


Peritoneal Carcinomatosis in Well-Differentiated Small-Intestinal Neuroendocrine Tumors with Mesenteric Tumor Deposits.
Well-differentiated small-intestinal neuroendocrine tumors (SI-NETs) tend to be biologically indolent. Despite this tendency, they have a predilection for metastasis. Peritoneal involvement is quite common as is unfortunately peritoneal carcinomatosis (PC). PC is a dreaded metastatic complication due to the significant morbidity it creates for patients as well as well as increasing their mortality risk. The risk factors for PC development in SI-NETs remain understudied; however, one such factor may be the presence of mesenteric tumor deposits (MTDs). We performed a retrospective analysis on 208 well-differentiated SI-NET patient samples, the majority with mesenteric masses, from the pathology archives of Vanderbilt University Medical Center. We sought to explore whether MTD presence was associated with PC, what other patient determinants were associated with PC and prognostic implication of these determinants. Patients with MTDs had an OR of 3.9 (CI 1.6, 10.9) for PC compared to patients without MTDs in the analysis. Patients who developed PC fared more poorly than those who did not (p=0.044). Our analysis, to the best of our knowledge, is the first to demonstrate an association between MTD presence and PC in this patient subgroup. We believe this finding warrants prospective evaluation given the possible therapeutic implications of being able to stratify SI-NET patients by their risk for developing PC based upon MTD presence.


Interleukin-1 gene cluster polymorphisms and susceptibility to clinical malaria in a Gambian case-control study.
Interleukin-1 (IL1) is a potent endogenous pyrogen and inducer of the acute phase response, and these innate immune responses are an important part of the human host's initial reaction to infection by the malaria parasite. In addition, several single-nucleotide polymorphisms (SNPs) in this region have previously been demonstrated to be associated with susceptibility to infectious disease. Therefore, a possible association with malaria susceptibility was investigated. A total of 13 polymorphic markers were used in a two-stage screening strategy to genotype a Gambian case-control study group by either restriction endonuclease digestion or the Sequenom MassARRAY assay. This involved an initial screen of 188 severe cases and 188 mild controls, and if there was a significant association with a malaria phenotype (P<0.05); this was followed by screening of the remaining 1044 samples. Two markers showed significant association with malaria: interleukin-1 alpha +4845 G --> T (P=0.035 for mild malaria versus controls) and interleukin-1 beta +3953 C --> T (P=0.030 for mild malaria versus severe malaria). Haplotypes constructed using the SNPHAP programme were not associated with any of the malaria phenotypes investigated. In summary, if IL1 variants are involved in malaria susceptibility in the Gambia at all, then the effects are small.


Manifestations of tuberculosis stigma within the healthcare system: the case of Sekondi-Takoradi Metropolitan district in Ghana.
To explore the manifestations of tuberculosis (TB) stigma within the healthcare system. Twenty-one individual interviews (16 females and 5 males) and six focus groups (1 with health managers, 1 with staff providing services for TB patients, and 4 with general staff) were conducted among healthcare workers (HCWs) and the generated data analysed using grounded theory principles and procedures. TB stigma manifests in three broad and inter-related ways within the healthcare system: behaviour towards TB patients; attitudes towards TB work; and administrative procedures and policies of healthcare mangers. Healthcare workers expressed fear of infection when interacting with TB patients; a fear which intensifies after the confirmation of the diagnosis. For fear of infection, they shuned, avoided, and advocated the segregation of TB patients at home and in the hospitals. They sometimes maltreated the patients, and accused and blamed them for deliberately infecting others. Posting to TB units/wards is viewed as a punishment, with majority indicating refusal to work there or be trained as TB nurse/doctor. They maintained that those working at TB units should be given incentives. Health managers also situated TB units/wards in isolated parts of the hospital, and do not provide adequate tools, equipment, support and supervision for those offering TB services. Fear of infection underlies the stigmatisation of TB by HCWs and worsen by administrative procedures and policies of healthcare managers. To help minimise TB stigma, a national guideline for the prevention of TB in HCWs should be developed as this may help protect them and alleviate their fear of infection. Rewarding and motivating HCWs involved in TB control may also improve their attitudes towards TB and those suffering from the disease. Furthermore, interventions to reduce TB stigma should be put in place, and the effects these may have on TB treatment outcomes investigated.


Diversity and community composition of methanogenic archaea in the rumen of Scottish upland sheep assessed by different methods.
Ruminal archaeomes of two mature sheep grazing in the Scottish uplands were analysed by different sequencing and analysis methods in order to compare the apparent archaeal communities. All methods revealed that the majority of methanogens belonged to the Methanobacteriales order containing the Methanobrevibacter, Methanosphaera and Methanobacteria genera. Sanger sequenced 1.3 kb 16S rRNA gene amplicons identified the main species of Methanobrevibacter present to be a SGMT Clade member Mbb. millerae (≥ 91% of OTUs); Methanosphaera comprised the remainder of the OTUs. The primers did not amplify ruminal Thermoplasmatales-related 16S rRNA genes. Illumina sequenced V6-V8 16S rRNA gene amplicons identified similar Methanobrevibacter spp. and Methanosphaera clades and also identified the Thermoplasmatales-related order as 13% of total archaea. Unusually, both methods concluded that Mbb. ruminantium and relatives from the same clade (RO) were almost absent. Sequences mapping to rumen 16S rRNA and mcrA gene references were extracted from Illumina metagenome data. Mapping of the metagenome data to 16S rRNA gene references produced taxonomic identification to Order level including 2-3% Thermoplasmatales, but was unable to discriminate to species level. Mapping of the metagenome data to mcrA gene references resolved 69% to unclassified Methanobacteriales. Only 30% of sequences were assigned to species level clades: of the sequences assigned to Methanobrevibacter, most mapped to SGMT (16%) and RO (10%) clades. The Sanger 16S amplicon and Illumina metagenome mcrA analyses showed similar species richness (Chao1 Index 19-35), while Illumina metagenome and amplicon 16S rRNA analysis gave lower richness estimates (10-18). The values of the Shannon Index were low in all methods, indicating low richness and uneven species distribution. Thus, although much information may be extracted from the other methods, Illumina amplicon sequencing of the V6-V8 16S rRNA gene would be the method of choice for studying rumen archaeal communities.


The effect of smoking and alcohol consumption on markers of systemic inflammation, immunoglobulin levels and immune response following pneumococcal vaccination in patients with arthritis.
The purpose of this research was to study the influence of cigarette smoking and alcohol consumption on immune response to heptavalent pneumococcal conjugate vaccine, immunoglobulin levels (Ig) and markers of systemic inflammation in patients with rheumatoid arthritis (RA) or spondylarthropathy (SpA). In total, 505 patients were vaccinated. Six pre-specified groups were enrolled: RA on methotrexate (MTX) treatment in some cases other disease-modifying antirheumatic drugs (DMARDs) (I); RA on anti-tumour necrosis factor (TNF) as monotherapy (II); RA on anti-TNF+MTX+ possibly other DMARDs (III); SpA on anti-TNF as monotherapy (IV); SpA on anti-TNF+MTX+ possibly other DMARDs (V); and SpA on nonsteroidal anti-inflammatory drugs (NSAIDs) and/or analgesics (VI). Smoking (pack-years) and alcohol consumption (g/week) were calculated from patient questionnaires. Ig, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were determined at vaccination. IgG antibodies against serotypes 23F and 6B were measured at vaccination and after four to six weeks using standard ELISA. Immune response (ratio between post- and pre-vaccination antibodies; immune response (IR)) and positive immune response (≥2-fold increase in pre-vaccination antibodies; posIR) were calculated. Eighty-eight patients (17.4%) were current smokers. Smokers had higher CRP and ESR, lower IgG and lower IR for both serotypes (P between 0.012 and 0.045). RA patients on MTX who smoked ≥1pack-year had lower posIR for both serotypes (P = 0.021; OR 0.29; CI 0.1 to 0.7) compared to never-smokers. Alcohol consumption was associated with lower CRP (P = 0.05) and ESR (P = 0.003) but did not influence IR or Ig levels. Smoking predicted impaired immune response to pneumococcal conjugate vaccine in RA patients on MTX. Smokers with arthritis had higher inflammatory markers and lower IgG regardless of diagnosis and treatment. Low to moderate alcohol consumption was related to lower levels of inflammation markers but had no impact on immune response. EudraCT EU 2007-006539-29 and NCT00828997.


Investing in children: changes in parental spending on children, 1972-2007.
Parental spending on children is often presumed to be one of the main ways that parents invest in children and a main reason why children from wealthier households are advantaged. Yet, although research has tracked changes in the other main form of parental investment-namely, time-there is little research on spending. We use data from the Consumer Expenditure Survey to examine how spending changed from the early 1970s to the late 2000s, focusing particularly on inequality in parental investment in children. Parental spending increased, as did inequality of investment. We also investigate shifts in the composition of spending and linkages to children's characteristics. Investment in male and female children changed substantially: households with only female children spent significantly less than parents in households with only male children in the early 1970s; but by the 1990s, spending had equalized; and by the late 2000s, girls appeared to enjoy an advantage. Finally, the shape of parental investment over the course of children's lives changed. Prior to the 1990s, parents spent most on children in their teen years. After the 1990s, however, spending was greatest when children were under the age of 6 and in their mid-20s.


Lactation Duration and Long-term Risk for Incident Type 2 Diabetes in Women With a History of Gestational Diabetes Mellitus.
We examined the association of lactation duration with incident type 2 diabetes among women with a history of gestational diabetes mellitus (GDM). We monitored 4,372 women with a history of GDM participating in the Nurses' Health Study II for incident type 2 diabetes over 25 years up to 2017. Lactation history was obtained through follow-up questionnaires to calculate lactation duration. Follow-up blood samples were collected from a subset of these women at median age of 58 years through the Diabetes & Women's Health Study. We documented 873 incident cases of type 2 diabetes during 87,411 person-years of follow-up. Longer duration of lactation was associated with lower risk of type 2 diabetes for both total lactation (hazard ratio 1.05 [95% CI 0.83-1.34] for up to 6 months, 0.91 [0.72-1.16] for 6-12 months, 0.85 [0.67-1.06] for 12-24 months, and 0.73 [0.57-0.93] for >24 months, compared with 0 months; P-trend = 0.003) and exclusive breastfeeding (P-trend = 0.002) after adjustment for age, ethnicity, family history of diabetes, parity, age at first birth, smoking, diet quality, physical activity, and prepregnancy BMI. Longer duration of lactation was also associated with lower HbA1c, fasting plasma insulin, and C-peptide concentrations among women without type 2 diabetes at follow-up (all adjusted P-trend ≤0.04). Longer duration of lactation is associated with a lower risk of type 2 diabetes and a favorable glucose metabolic biomarker profile among women with a history of GDM. The underlying mechanisms and impact on diabetes complications, morbidity, and mortality remain to be determined.


The final step of hygromycin A biosynthesis, oxidation of C-5''-dihydrohygromycin A, is linked to a putative proton gradient-dependent efflux.
Hygromycin A (HA) is an aminocyclitol antibiotic produced and excreted by Streptomyces hygroscopicus. Deletion of hyg26 from the hygromycin A biosynthetic gene cluster has previously been shown to result in a mutant that produces 5''-dihydrohygromycin A (DHHA). We report herein on the purification and characterization of Hyg26 expressed in Escherichia coli. The enzyme catalyzes an NAD(H)-dependent reversible interconversion of HA and DHHA, supporting the role of the reduced HA as the penultimate biosynthetic pathway intermediate and not a shunt product. The equilibrium for the Hyg26-catalyzed reaction heavily favors the DHHA intermediate. The high-titer production of the HA product by S. hygroscopicus must be dependent upon a subsequent energetically favorable enzyme-catalyzed process, such as the selective and efficient export of HA. hyg19 encodes a putative proton gradient-dependent transporter, and a mutant lacking this gene was observed to produce less HA and to produce the DHHA intermediate. The DHHA produced by either the Deltahyg19 or the Deltahyg26 mutant had slightly reduced activity against E. coli and reduced protein synthesis-inhibitory activity in vitro. The data indicate that Hyg26 and Hyg19 have evolved to produce and export the final potent HA product in a coordinated fashion.


Conflict in medical teams: opportunity or danger?
Intragroup conflicts often occur when people are called upon to collaborate in the accomplishment of a task. For example, when surgeons and nurses work together during an operation, conflicts may emerge because of differences in functional understanding. Whether these conflicts are beneficial or detrimental to team outcomes has been the source of much debate. From one perspective, a conflict that stems from differences in members' functional understanding may enhance team members' understanding and performance of the task at hand. By contrast, such a conflict may cause hostility, emotionality and distraction from actual task accomplishment. This study reviews findings on the relationships between intragroup conflict and team outcomes, discusses potential conflict resolution strategies for intragroup conflicts and explores how these link to the field of medical education. Three primary types of conflict have been distinguished, involving, respectively, task-, process- and relationship-associated conflict. Both process conflict, or conflict about the logistics of task accomplishment, and relationship conflict, or conflict about interpersonal incompatibilities, have been shown to detract from effective team functioning. Task conflict, or conflict about the content of the task itself, is also generally negative for team functioning, but under certain conditions its negative effects may be minimised. For example, when teams can clearly separate task issues from relationship issues, task conflicts are less destructive for team outcomes. However, achieving such a separation in practice, and thereby realising the benefits of task conflict, is quite difficult to achieve. Intragroup conflicts pose a challenge to effective team functioning. In the education of medical professionals, effective training in conflict management skills and their application to specific team conflict dynamics, such as with reference to how to resolve task as opposed to relationship conflict, is critical.


Single-cell genomics based on Raman sorting reveals novel carotenoid-containing bacteria in the Red Sea.
Cell sorting coupled with single-cell genomics is a powerful tool to circumvent cultivation of microorganisms and reveal microbial 'dark matter'. Single-cell Raman spectra (SCRSs) are label-free biochemical 'fingerprints' of individual cells, which can link the sorted cells to their phenotypic information and ecological functions. We employed a novel Raman-activated cell ejection (RACE) approach to sort single bacterial cells from a water sample in the Red Sea based on SCRS. Carotenoids are highly diverse pigments and play an important role in phototrophic bacteria, giving strong and distinctive Raman spectra. Here, we showed that individual carotenoid-containing cells from a Red Sea sample were isolated based on the characteristic SCRS. RACE-based single-cell genomics revealed putative novel functional genes related to carotenoid and isoprenoid biosynthesis, as well as previously unknown phototrophic microorganisms including an unculturable Cyanobacteria spp. The potential of Raman sorting coupled to single-cell genomics has been demonstrated.


Long non-coding RNAs coordinate cellular responses to stress.
Following the initial discovery of the heat shock RNA omega (hsrω) gene of Drosophila melanogaster to be non-coding (nc) and also inducible by cell stress, other stress-inducible long non-coding RNAs (lncRNA) have been described in diverse organisms. In view of the rapid sequence divergence of lncRNAs, present knowledge of stress trasncriptome is limited and fragmented. Several known stress-related lncRNAs, associated with specific nuclear speckled domains or nucleolus, provide structural base for sequestering diverse RNA-processing/regulatory proteins. Others have roles in transcriptional or translational inhibition during stress or in signaling pathways; functions of several other lncRNAs are not yet known. Most stress-related lncRNAs act primarily by modulating activity of the proteins to which they bind or by sequestering specific sets of proteins away from the active pool. A common emerging theme is that a given lncRNA targets one or more protein/s with key role/s in the cascade of events triggered by the stress and therefore has a widespread integrative effect. Since proteins associate with RNA through short sequence motifs, the overall base sequence of functionally similar ncRNAs is often not conserved except for specific motifs. The rapid evolvability of ncRNA sequences provides elegant modules for adaptability to changing environment as binding of one or the other protein to ncRNA can alter its structure and functions in distinct ways. Thus the stress-related lncRNAs act as hubs in the cellular networks to coordinate activities of the members within and between different networks to maintain cellular homeostasis for survival or to trigger cell death.


Synthesis, Characterization, Antimicrobial and Antitumor Activities of Sucrose Octa(N-ethyl)carbamate.
Sucrose octa(N-ethyl)carbamate was synthesized directly from sucrose and ethyl isocyanate, and its structure was confirmed by various analytical methods, such as (1)H and (13)C NMR, FTIR, m.p., MS, and optical rotation. Its antibacterial, antifungal and cytotoxic activities were investigated. It exhibited strong inhibition against all bacteria tested, namely S. aureus (MIC 0.18±0.006), B. cereus (MIC 0.094±0.000), M. flavus (MIC 0.28±0.01), L. monocytogenes (MIC 0.18±0.006), P. aeruginosa (MIC 0.094±0.002), S. typhimurium (MIC 0.094±0.002), E. coli (MIC 0.18±0.006) and E. cloacae (MIC 0.18±0.006) and strong antifungal activity towards T. viride (MIC 0.09 ± 0.006), A. versicolor (MIC 0.18 ± 0.01), A. ochraceus (MIC 0.375 ± 0.01) and P. ochrochloron (MIC 0.375 ± 0.04). Furthermore, it showed moderate antitumor potential against human breast (GI50 357.20±14.12), colon (GI50 332.43±11.19) and cervical (GI50 282.67±3.97) cell lines and, more important, without hepatotoxicity.


The chondrogenic differentiation potential of dental pulp stem cells.
Dental pulp stem cells (DPSCs) are particularly promising for tissue engineering (TE) due to the ease of their isolation procedure, great expansion potential and capability to differentiate towards several cell types of the mesodermal, ectodermal and endodermal lineages. Although several studies hint that DPSCs exhibit potential for cartilage tissue formation, the chondrogenic potential of DPSCs has only been marginally explored. Thus, the aim of the present study was to closely investigate the chondrogenic differentiation capacity of DPSCs for TE applications. More specifically, the potential of DPSCs for engineering hyaline and fibrous cartilage was determined. DPSCs obtained from 7 human molars were expanded and chondrogenically differentiated in a 3D pellet culture model. After 21 d of differentiation with chondrogenic stimuli, DPSCs displayed glycosaminoglycan, aggrecan and limited collagen type II deposition. Cells presented an elongated morphology and produced a collagen-rich extracellular matrix, with a predominance of collagen type I in most of the samples, a characteristic of fibrous cartilage tissue. Variations in the administration periods of several chondro-inductive growth factors, including transforming growth factor beta 3, bone morphogenetic protein-2, -6, -7 and insulin-like growth factor-1, did not increase glycosaminoglycan or collagen type II deposition, typical markers of hyaline cartilage tissue. Furthermore, DPSCs could not be stimulated to go into hypertrophic chondrogenesis. These results indicated that under a large variety of chondro-inductive culture conditions, DPSCs could form fibrocartilaginous tissues but not hyaline cartilage. Thus, DPSCs represent a valuable cell source for the regeneration of fibrocartilage in joints.


Measurement of the Absolute Magnitude and Time Courses of Mitochondrial Membrane Potential in Primary and Clonal Pancreatic Beta-Cells.
The aim of this study was to simplify, improve and validate quantitative measurement of the mitochondrial membrane potential (ΔψM) in pancreatic β-cells. This built on our previously introduced calculation of the absolute magnitude of ΔψM in intact cells, using time-lapse imaging of the non-quench mode fluorescence of tetramethylrhodamine methyl ester and a bis-oxonol plasma membrane potential (ΔψP) indicator. ΔψM is a central mediator of glucose-stimulated insulin secretion in pancreatic β-cells. ΔψM is at the crossroads of cellular energy production and demand, therefore precise assay of its magnitude is a valuable tool to study how these processes interplay in insulin secretion. Dispersed islet cell cultures allowed cell type-specific, single-cell observations of cell-to-cell heterogeneity of ΔψM and ΔψP. Glucose addition caused hyperpolarization of ΔψM and depolarization of ΔψP. The hyperpolarization was a monophasic step increase, even in cells where the ΔψP depolarization was biphasic. The biphasic response of ΔψP was associated with a larger hyperpolarization of ΔψM than the monophasic response. Analysis of the relationships between ΔψP and ΔψM revealed that primary dispersed β-cells responded to glucose heterogeneously, driven by variable activation of energy metabolism. Sensitivity analysis of the calibration was consistent with β-cells having substantial cell-to-cell variations in amounts of mitochondria, and this was predicted not to impair the accuracy of determinations of relative changes in ΔψM and ΔψP. Finally, we demonstrate a significant problem with using an alternative ΔψM probe, rhodamine 123. In glucose-stimulated and oligomycin-inhibited β-cells the principles of the rhodamine 123 assay were breached, resulting in misleading conclusions.


How relationship status and sociosexual orientation influence the link between facial attractiveness and visual attention.
Facial attractiveness captures and binds visual attention, thus affecting visual exploration of our environment. It is often argued that this effect on attention has evolutionary functions related to mating. Although plausible, such perspectives have been challenged by recent behavioral and eye-tracking studies, which have shown that the effect on attention is moderated by various sex- and goal-related variables such as sexual orientation. In the present study, we examined how relationship status and sociosexual orientation moderate the link between attractiveness and visual attention. We hypothesized that attractiveness leads to longer looks and that being single as well as being more sociosexually unrestricted, enhances the effect of attractiveness. Using an eye-tracking free-viewing paradigm, we tested 150 heterosexual men and women looking at images of urban real-world scenes depicting two people differing in facial attractiveness. Participants additionally provided attractiveness ratings of all stimuli. We analyzed the correlations between how long faces were looked at and participants' ratings of attractiveness and found that more attractive faces-especially of the other sex-were looked at longer. We also found that more sociosexually unrestricted participants who were single had the highest attractiveness-attention correlation. Our results show that evolutionary predictions cannot fully explain the attractiveness-attention correlation; perceiver characteristics and motives moderate this relationship.


Health care need and health disparities: Findings from the Regional South Australia Health (RESONATE) survey.
Regional South Australia has some of the highest rates of psychological distress, chronic disease and multimorbidity of any Australian State or Territory. Yet, the healthcare needs of this population are still not completely understood. To better understand the healthcare needs of regional South Australians, we invited adults living in the region to complete the 44-item consumer utilisation, expectations and experiences of healthcare instrument (CONVERSATIONS), online or in hard-copy. The survey was conducted between April 2017 and March 2018. A multi-modal recruitment campaign was utilised to promote the survey. We examined associations between study outcomes and remoteness area, and drew comparisons between our findings and other surveys reporting pertinent outcomes in the urban SA population. The questionnaire was completed by 3,926 adults (52.5% females; 37.6% aged 60 + years). Among the 264 distinct health conditions reported by participants, the most prevalent were hypertension (31.6%), depression (25.7%), anxiety (23.5%) and hypercholesterolaemia (22.9%). The lifetime prevalence of these conditions among participants exceeded rates reported in urban SA. The largest regional-urban health disparities were observed for eczema/dermatitis, skin cancer, other cancer types and cataracts, where prevalence rates were 2075%, 400%, 373% and 324% higher, respectively, than that reported in urban SA. Participants also reported higher levels of multimorbidity (37.7% higher) relative to urban South Australians. By contrast, participants appeared to be exposed to fewer lifestyle risk behaviours (e.g. smoking, alcohol, inadequate fruit or vegetable intake) than their urban counterparts. In summary, there was a high level of healthcare need, and considerable health disparity among participants when compared with urban settings (particularly for skin and eye conditions). These findings highlight the need for a more targeted approach to delivering health services and health promotion activities in regional areas.


Identification of rpo30, a vaccinia virus RNA polymerase gene with structural similarity to a eucaryotic transcription elongation factor.
Eucaryotic transcription factors that stimulate RNA polymerase II by increasing the efficiency of elongation of specifically or randomly initiated RNA chains have been isolated and characterized. We have identified a 30-kilodalton (kDa) vaccinia virus-encoded protein with apparent homology to SII, a 34-kDa mammalian transcriptional elongation factor. In addition to amino acid sequence similarities, both proteins contain C-terminal putative zinc finger domains. Identification of the gene, rpo30, encoding the vaccinia virus protein was achieved by using antibody to the purified viral RNA polymerase for immunoprecipitation of the in vitro translation products of in vivo-synthesized early mRNA selected by hybridization to cloned DNA fragments of the viral genome. Western immunoblot analysis using antiserum made to the vaccinia rpo30 protein expressed in bacteria indicated that the 30-kDa protein remains associated with highly purified viral RNA polymerase. Thus, the vaccinia virus protein, unlike its eucaryotic homolog, is an integral RNA polymerase subunit rather than a readily separable transcription factor. Further studies showed that the expression of rpo30 is regulated by dual early and later promoters.


Do female age and body weight modify the effect of individualized FSH dosing in IVF/ICSI treatment? A secondary analysis of the OPTIMIST trial.
The OPTIMIST trial revealed that for women starting in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment, no substantial differences exist in first cycle and cumulative live birth rates between an antral follicle count (AFC)-based individualized follicle-stimulating hormone (FSH) dose and a standard dose. Female age and body weight have been suggested to cause heterogeneity in the effect of FSH dose individualization. The objective of the current study is to evaluate whether these patient characteristics modify the effect of AFC-based individualized FSH dosing in IVF/ICSI treatment. A secondary data-analysis of the OPTIMIST trial. Women initiating IVF/ICSI treatment were classified as predicted poor (AFC 0-7), suboptimal (AFC 8-10) or hyper responders (AFC >15), and randomly allocated to a standard FSH dose (150 IU/d) or an individualized FSH dose (450, 225 or 100 IU/d for predicted poor, suboptimal and hyper responders, respectively). In each predicted response category, logistic regression models with interaction terms were used to evaluate the presence of effect modification. The first cycle was analyzed, and the primary outcomes were first complete cycle live birth rate (including fresh plus frozen-thawed embryo transfers) and ovarian hyperstimulation syndrome (OHSS) risks. No effect modification was revealed in the predicted poor (n = 234) and suboptimal (n = 277) responders. In the predicted hyper responders (n = 521), the effect of the individualized FSH dose on the first cycle live birth rate was modified by female age (P = 0.02) and the effect on OHSS risks was modified by body weight (P = 0.02). A dose reduction from 150 to 100 IU/d generally decreased the OHSS risks in predicted hyper responders, but also reduced the chance of a live birth in young women, and had no beneficial impact on OHSS risks in women with a relatively low body weight. In women with a predicted hyper response undergoing IVF/ICSI treatment, female age and body weight seem to modify the effect of FSH dose individualization. Although a reduced FSH starting dose generally decreases the OHSS risks, it may also reduce the chance of a live birth, specifically for young women. Future studies could consider these findings when investigating the optimal approach to reduce OHSS risks while maintaining the probability of a live birth for predicted hyper responders in IVF/ICSI treatment.


Exploring the Role of "Brahmi" (Bacopa monnieri and Centella asiatica) in Brain Function and Therapy
It has been envisaged that in this century, disorders of the central nervous system will have a significant bearing on the healthcare concerns of the human population worldwide. Such neurological and psychiatric disorders are generally associated with loss of memory, cognitive deficits, impaired mental function etc. Due to the multi-factorial nature of these diseases, modern medicine based psychoactive drugs have met with limited success. Therefore, there is a growing demand for novel products that could target multiple pathways and improve the mental capabilities either independently or in combination with conventional drugs. In the recent times, herbal products based on traditional knowledge have been increasingly used both in developed and developing countries. According to "Ayurveda", the Indian traditional system of medicine, "medhyarasayanas" represent herbal therapeutics that boost memory, restore cognitive deficits and improve mental function. The current review deals with the components and application of such a traditional herb "Brahmi" that corresponds to two plants, Bacopa monnieri and Centella asiatica. Research evidences clearly indicate that both plants possess neuroprotective properties, have nootropic activity with therapeutic implications for patients with memory loss. The field has witnessed exciting patent activity with most inventions aiming at either (i) improving the methods of herbal extraction or (ii) enrichment and purification of novel compounds from brahmi or (iii) providing novel synergistic formulations for therapeutics in various human ailments. In this review, clinical trials related to the therapeutic properties of brahmi and current patents relevant to the preparation, composition and application have also been included.


Evaluation of the host-specificity and prevalence of enterococci surface protein (esp) marker in sewage and its application for sourcing human fecal pollution.
The suitability of the enterococci surface protein (esp) marker to detect human fecal pollution was evaluated by testing 197 fecal samples from 13 host groups in Southeast Queensland, Australia. Overall, this marker was detected in 90.5% of sewage and septic system samples and could not be detected in any fecal samples from 12 animal host groups. The sensitivity of the esp primer to detect the human-specific esp marker in sewage and septic samples was 100 and 67%, respectively. The overall specificity of this marker to distinguish between human and animal fecal pollution was 100%. Its prevalence in sewage was also determined by testing samples from the raw sewage, secondary effluent, and treated effluent of a sewage treatment plant (STP) over five consecutive days. Of the 15 samples tested, 12 (80%) were found to be positive for this marker. In contrast, it was not found in three samples from the treated effluent and these samples did not contain any culturable enterococci. The PCR limit of detection of this marker in freshwater samples was up to dilution 1 x 10(-4) and the number of culturable enterococci at this dilution was 4.8 x 10(1) +/- 7.0 x 10 degrees colony forming unit (CFU). The utility of this marker was evaluated by testing water samples from three non-sewered catchments in Pine Rivers in Southeast Queensland. Of the 13 samples tested, eight were positive for this marker with the number of enterococci ranging between 1.8 x 10(3) to 8.5 x 10(3) CFU per 100 mL of water. Based on the results, it can be concluded that the esp marker appears to be sewage specific and could be used as a reliable marker to detect human fecal pollution in surface waters in Southeast Queensland, Australia.


Active Ingredients and Mechanisms of Change in Motivational Interviewing for Smoking Cessation in Patients With Coronary Artery Disease: A Mixed Methods Study.
Background: For patients with coronary artery disease (CAD), smoking is an important risk factor for the recurrence of a cardiovascular event. Motivational interviewing (MI) may increase the motivation of the smokers to stop smoking. Data on MI for smoking cessation in patients with CAD are limited, and the active ingredients and working mechanisms of MI in smoking cessation are largely unknown. Therefore, this study was designed to explore active ingredients and working mechanisms of MI for smoking cessation in smokers with CAD, shortly after a cardiovascular event. Methods: We conducted a qualitative multiple case study of 24 patients with CAD who participated in a randomized trial on lifestyle change. One hundred and nine audio-recorded MI sessions were coded with a combination of the sequential code for observing process exchanges (SCOPE) and the motivational interviewing skill code (MISC). The analysis of the cases consisted of three phases: single case analysis, cross-case analysis, and cross-case synthesis. In a quantitative sequential analysis, we calculated the transition probabilities between the use of MI techniques by the coaches and the subsequent patient statements concerning smoking cessation. Results: In 12 cases, we observed ingredients that appeared to activate the mechanisms of change. Active ingredients were compositions of behaviors of the coaches (e.g., supporting self-efficacy and supporting autonomy) and patient reactions (e.g., in-depth self-exploration and change talk), interacting over large parts of an MI session. The composition of active ingredients differed among cases, as the patient process and the MI-coaching strategy differed. Particularly, change talk and self-efficacy appeared to stimulate the mechanisms of change "arguing oneself into change" and "increasing self-efficacy/confidence." Conclusion: Harnessing active ingredients that target the mechanisms of change "increasing self-efficacy" and "arguing oneself into change" is a good MI strategy for smoking cessation, because it addresses the ambivalence of a patient toward his/her ability to quit, while, after the actual cessation, maintaining the feeling of urgency to persist in not smoking in the patient.


Deep RNA sequencing analysis of readthrough gene fusions in human prostate adenocarcinoma and reference samples.
Readthrough fusions across adjacent genes in the genome, or transcription-induced chimeras (TICs), have been estimated using expressed sequence tag (EST) libraries to involve 4-6% of all genes. Deep transcriptional sequencing (RNA-Seq) now makes it possible to study the occurrence and expression levels of TICs in individual samples across the genome. We performed single-end RNA-Seq on three human prostate adenocarcinoma samples and their corresponding normal tissues, as well as brain and universal reference samples. We developed two bioinformatics methods to specifically identify TIC events: a targeted alignment method using artificial exon-exon junctions within 200,000 bp from adjacent genes, and genomic alignment allowing splicing within individual reads. We performed further experimental verification and characterization of selected TIC and fusion events using quantitative RT-PCR and comparative genomic hybridization microarrays. Targeted alignment against artificial exon-exon junctions yielded 339 distinct TIC events, including 32 gene pairs with multiple isoforms. The false discovery rate was estimated to be 1.5%. Spliced alignment to the genome was less sensitive, finding only 18% of those found by targeted alignment in 33-nt reads and 59% of those in 50-nt reads. However, spliced alignment revealed 30 cases of TICs with intervening exons, in addition to distant inversions, scrambled genes, and translocations. Our findings increase the catalog of observed TIC gene pairs by 66%.We verified 6 of 6 predicted TICs in all prostate samples, and 2 of 5 predicted novel distant gene fusions, both private events among 54 prostate tumor samples tested. Expression of TICs correlates with that of the upstream gene, which can explain the prostate-specific pattern of some TIC events and the restriction of the SLC45A3-ELK4 e4-e2 TIC to ERG-negative prostate samples, as confirmed in 20 matched prostate tumor and normal samples and 9 lung cancer cell lines. Deep transcriptional sequencing and analysis with targeted and spliced alignment methods can effectively identify TIC events across the genome in individual tissues. Prostate and reference samples exhibit a wide range of TIC events, involving more genes than estimated previously using ESTs. Tissue specificity of TIC events is correlated with expression patterns of the upstream gene. Some TIC events, such as MSMB-NCOA4, may play functional roles in cancer.


[The impact of antimicrobial resistance and Streptococcus pneumoniae serotype distribution on the mortality of children under 5 years of age with invasive disease].
Severe pneumonia and meningitis caused by Streptococcus pneumoniae have been persistently associated with high mortality rates, despite advances in antimicrobial therapy and the development of vaccines. Resistance to penicillin and other antimicrobial agents is increasing and spreading worldwide. Even though risk factors for development of antimicrobial resistance have been identified, their influence on mortality has not been clarified. With regard to virulence, differences among serotypes have been determined, but their impact on mortality is unknown. The aim of this study was to determine the risk factors associated with mortality in children with invasive pneumococcal disease. Clinical records for 245 children under 5 years of age with invasive disease due to S.pneumoniae were reviewed. Children were diagnosed between 1994 and 1996 in Colombia, during the study of S.pneumoniae capsular types conducted by the Pan American Health Organization's Regional System for Vaccines. Of the 245 patients whose charts were examined, 29 (11%) died. No significant differences in age, gender, underlying disease, nor antimicrobial treatment concordance were found. Variables associated with mortality in the univariate analysis were a diagnosis of meningitis; antimicrobial resistance to penicillin, trimethoprim-sulfamethoxazole (TMS), or erythromycin; multiresistance, and serotypes 6, 23F, 7F, 8, and 35B. In the logistic regression, serotypes 7F (OR = 7.13; P = 0.04) and 8 (OR = 13.8; P = 0.07), polipnea (OR = 2.74; P = 0.03), meningitis (OR = 5.02; P = 0.0001) and TMS resistance (OR = 2.62; P = 0.02) continued to be associated with mortality. In patients with pneumonia, serotype was the factor most consistently associated with mortality; in meningitis patients, it was antimicrobial resistance. Differences in mortality according to serotype must be taken into account in developing a vaccine if a substantial impact on pneumococcal disease morbidity and mortality is to be achieved.


The effects of ketamine and risperidone on eye movement control in healthy volunteers.
The non-competitive N-methyl-D-aspartate receptor antagonist ketamine leads to transient psychosis-like symptoms and impairments in oculomotor performance in healthy volunteers. This study examined whether the adverse effects of ketamine on oculomotor performance can be reversed by the atypical antipsychotic risperidone. In this randomized double-blind, placebo-controlled study, 72 healthy participants performed smooth pursuit eye movements (SPEM), prosaccades (PS) and antisaccades (AS) while being randomly assigned to one of four drug groups (intravenous 100 ng ml(-1) ketamine, 2 mg oral risperidone, 100 ng ml(-1) ketamine plus 2 mg oral risperidone, placebo). Drug administration did not lead to harmful adverse events. Ketamine increased saccadic frequency and decreased velocity gain of SPEM (all P < 0.01) but had no significant effects on PS or AS (all P > or = 0.07). An effect of risperidone was observed for amplitude gain and peak velocity of PS and AS, indicating hypometric gain and slower velocities compared with placebo (both P < or = 0.04). No ketamine by risperidone interactions were found (all P > or = 0.26). The results confirm that the administration of ketamine produces oculomotor performance deficits similar in part to those seen in schizophrenia. The atypical antipsychotic risperidone did not reverse ketamine-induced deteriorations. These findings do not support the cognitive enhancing potential of risperidone on oculomotor biomarkers in this model system of schizophrenia and point towards the importance of developing alternative performance-enhancing compounds to optimise pharmacological treatment of schizophrenia.


Nuclear receptor coactivator 6 mediates the synergistic activation of human cytochrome P-450 2C9 by the constitutive androstane receptor and hepatic nuclear factor-4alpha.
Nuclear receptor coactivator 6 (NCOA6) also known as PRIP/RAP250/ASC-2 anchors a steady-state complex of cofactors and function as a transcriptional coactivator for certain nuclear receptors. This is the first study to identify NCOA6 as a hepatic nuclear factor 4alpha (HNF4alpha)-interacting protein. CYP2C9 is an important enzyme that metabolizes both commonly used therapeutic drugs and important endogenous compounds. We have shown previously that constitutive androstane receptor (CAR) (a xenobiotic-sensing receptor) up-regulates the CYP2C9 promoter through binding to a distal site, whereas HNF4alpha transcriptionally up-regulates CYP2C9 via proximal sites. We demonstrate ligand-enhanced synergistic cross-talk between CAR and HNF4alpha. We identify NCOA6 as crucial to the underlying mechanism of this cross-talk. NCOA6 was identified as an HNF4alpha-interacting protein in this study using a yeast two-hybrid screen and GST pull-down assays. Furthermore, we identified NCOA6, CAR, and other coactivators as part of a mega complex of cofactors associated with HNF4alpha in HepG2 cells. Although the interaction of NCOA6 with CAR is specifically through the first LXXLL motif of NCOA6, both LXXLL motifs are involved in its interaction with HNF4alpha. Silencing of NCOA6 abrogated the synergistic activation of the CYP2C9 promoter and the synergistic induction of the CYP2C9 gene by CAR-HNF4alpha. Chromatin immunoprecipitation analysis revealed that NCOA6 can pull down both the proximal HNF4alpha and distal CAR binding sites of the CYP2C9 promoter and provides the basis for the recruitment of other cofactors. We conclude that the coactivator NCOA6 mediates the mechanism of the synergistic activation of the CYP2C9 gene by CAR and HNF4alpha.


Bacteriological and molecular analysis of rifampin-resistant Mycobacterium tuberculosis strains isolated in Australia.
To develop a better understanding of the epidemiology and molecular biology of rifampin-resistant Mycobacterium tuberculosis strains in Australia, 50 clinical isolates (33 rifampin-resistant and 17 rifampin-sensitive strains) cultured between 1990 and 1997 were analyzed by a number of bacteriological and molecular techniques. Examination of the drug resistance profiles of the 33 rifampin-resistant isolates revealed that 91% were resistant to rifampin in combination with resistance to isoniazid, 88% were resistant to rifampin on first isolation, and 81% showed cross-resistance with rifabutin. On the basis of the demographic data provided for the patients infected with the rifampin-resistant strains, 90% of the patients were born overseas. Of these patients, 64% developed clinical symptoms within 5 years of residence in Australia. On a molecular level, analysis of the rpoB gene revealed that 97% of the rifampin-resistant isolates had missense mutations within a conserved region of the gene, and eight types of missense mutations were detected. Of the 31 rifampin-resistant isolates that were typed by restriction fragment length polymorphism (RFLP) analysis, 28 distinct patterns were obtained by RFLP analysis with IS6110, and three clusters of genetically related isolates were identified. All isolates within the clusters were from patients who were born overseas and who had the same country of origin. The results from this study provide an overview of the current situation of rifampin resistance in Australia and can serve as a basis for continued monitoring of drug-resistant M. tuberculosis strains isolated within the country.


A national facility for biological cryo-electron microscopy.
Three-dimensional electron microscopy is an enormously powerful tool for structural biologists. It is now able to provide an understanding of the molecular machinery of cells, disease processes and the actions of pathogenic organisms from atomic detail through to the cellular context. However, cutting-edge research in this field requires very substantial resources for equipment, infrastructure and expertise. Here, a brief overview is provided of the plans for a UK national three-dimensional electron-microscopy facility for integrated structural biology to enable internationally leading research on the machinery of life. State-of-the-art equipment operated with expert support will be provided, optimized for both atomic-level single-particle analysis of purified macromolecules and complexes and for tomography of cell sections. The access to and organization of the facility will be modelled on the highly successful macromolecular crystallography (MX) synchrotron beamlines, and will be embedded at the Diamond Light Source, facilitating the development of user-friendly workflows providing near-real-time experimental feedback.


Diverse convergent evidence in the genetic analysis of complex disease: coordinating omic, informatic, and experimental evidence to better identify and validate risk factors.
In omic research, such as genome wide association studies, researchers seek to repeat their results in other datasets to reduce false positive findings and thus provide evidence for the existence of true associations. Unfortunately this standard validation approach cannot completely eliminate false positive conclusions, and it can also mask many true associations that might otherwise advance our understanding of pathology. These issues beg the question: How can we increase the amount of knowledge gained from high throughput genetic data? To address this challenge, we present an approach that complements standard statistical validation methods by drawing attention to both potential false negative and false positive conclusions, as well as providing broad information for directing future research. The Diverse Convergent Evidence approach (DiCE) we propose integrates information from multiple sources (omics, informatics, and laboratory experiments) to estimate the strength of the available corroborating evidence supporting a given association. This process is designed to yield an evidence metric that has utility when etiologic heterogeneity, variable risk factor frequencies, and a variety of observational data imperfections might lead to false conclusions. We provide proof of principle examples in which DiCE identified strong evidence for associations that have established biological importance, when standard validation methods alone did not provide support. If used as an adjunct to standard validation methods this approach can leverage multiple distinct data types to improve genetic risk factor discovery/validation, promote effective science communication, and guide future research directions.


Prevalence and typing of Listeria monocytogenes in raw catfish fillets.
Raw channel catfish fillets collected from three processing plants during four time periods were tested for the presence of Listeria species. Listeria monocytogenes was the predominant Listeria species found in these catfish fillets, with 25 to 47% prevalence. Other Listeria species, such as L. welshimeri, L. innocua, L. ivanovii, L. grayi, and L. seeligeri, were also found. L. monocytogenes isolates were further fingerprinted by a repetitive element PCR. Forty distinctive electrophoretic types (ETs) and three genetic clusters were determined by Dice coefficient analysis and UPGMA (unweighted pair group method using arithmetic averages). Twenty of 40 ETs were represented by a single isolate, and the other 20 ETs were represented by 2 to 11 isolates. Thirty-five ETs, represented by 76 isolates, were found in processing plant A, B, or C and designated plant-specific types. The remaining five ETs, represented by 21 isolates, were found in multiple plants and designated nonplant-specific types. In addition, 10 ETs from 52 isolates were found repeatedly during different seasons. Plant-specific and nonplant-specific L. monocytogenes coexisted in processed catfish fillets. Some isolates were persistently found in processed fillets, suggesting that either the current sanitation procedures used by these plants are inadequate or that these isolates originated from the natural habitats of the catfish. The results also suggest that the repetitive element PCR is a useful tool for differentiating L. monocytogenes subtypes and can be used for tracing the source of a contamination.


Detection of pelvic lymph node micrometastasis in stage IA2-IB2 cervical cancer by immunohistochemical analysis.
The objectives of this study were to (1) determine the incidence of lymph node micrometastasis in cervical cancer by immunohistochemical analysis and (2) determine if the presence of micrometastasis is a poor prognostic feature in early cervical cancer. We retrospectively reviewed the medical records of 62 patients who underwent radical hysterectomy and lymphadenectomy for FIGO stage IA2-IB2 cervical cancer at Stanford University Hospital from 1990 to 2000. Forty-nine patients with negative lymph nodes were identified. A total of 976 formalin-fixed paraffin-embedded pelvic lymphadenectomy specimens were serially sectioned and stained with anti-cytokeratin antibodies AE1 and AE1/CAM5.2. Six patients had stage IA2 disease, 37 had stage IB1, and 6 had IB2. The mean age of the patients was 44 years (range, 24-76). Seventy-one percent had squamous cell carcinomas, 22% had adenocarcinomas, and 6% had other types. Lymph node micrometastases were immunohistochemically detected in 4 of the 49 (8.1%) patients, comprising 4 of 976 (0.41%) pelvic lymph nodes examined. Twelve of 45 (15.6%) patients with negative nodes had lymph-vascular space invasion (LVSI) whereas 3 of 4 (75%) patients with micrometastases had LVSI. At a mean follow-up time of 39.4 months, 2 of 4 (50%) patients with micrometastasis had recurrent disease, while 3 of 45 (6.7%) patients without micrometastasis developed recurrent disease. These preliminary data suggest that immunohistochemical detection of pelvic lymph nodes is more frequent in patients with LVSI and may identify patients needing adjuvant chemoradiation.


The AINTEGUMENTA LIKE1 homeotic transcription factor PtAIL1 controls the formation of adventitious root primordia in poplar.
Adventitious rooting is an essential but sometimes rate-limiting step in the clonal multiplication of elite tree germplasm, because the ability to form roots declines rapidly with age in mature adult plant tissues. In spite of the importance of adventitious rooting, the mechanism behind this developmental process remains poorly understood. We have described the transcriptional profiles that are associated with the developmental stages of adventitious root formation in the model tree poplar (Populus trichocarpa). Transcriptome analyses indicate a highly specific temporal induction of the AINTEGUMENTA LIKE1 (PtAIL1) transcription factor of the AP2 family during adventitious root formation. Transgenic poplar samples that overexpressed PtAIL1 were able to grow an increased number of adventitious roots, whereas RNA interference mediated the down-expression of PtAIL1 expression, which led to a delay in adventitious root formation. Microarray analysis showed that the expression of 15 genes, including the transcription factors AGAMOUS-Like6 and MYB36, was overexpressed in the stem tissues that generated root primordia in PtAIL1-overexpressing plants, whereas their expression was reduced in the RNA interference lines. These results demonstrate that PtAIL1 is a positive regulator of poplar rooting that acts early in the development of adventitious roots.


Functional disconnection of a prefrontal cortical-dorsal striatal system disrupts choice reaction time performance: implications for attentional function.
This series of experiments investigated the role of a prefrontal cortical-dorsal striatal circuit in attention, using a continuous performance task of sustained and spatially divided visual attention. A unilateral excitotoxic lesion of the medial prefrontal cortex and a contralateral lesion of the medial caudate-putamen were used to "disconnect" the circuit. Control groups of rats with unilateral lesions of either structure were tested in the same task. Behavioral controls included testing the effects of the disconnection lesion on Pavlovian discriminated approach behavior. The disconnection lesion produced a significant reduction in the accuracy of performance in the attentional task but did not impair Pavlovian approach behavior or affect locomotor or motivational variables, providing evidence for the involvement of this medial prefrontal corticostriatal system in aspects of visual attentional function.


Tetraethylthiuram disulfide (Antabuse) inhibits the human malaria parasite Plasmodium falciparum.
Plasmodium falciparum in culture grows optimally at 3% oxygen. Oxygen levels down to 0.5% still support growth, but anaerobic conditions do not. These findings, and the absence of the Krebs cycle in Plasmodium, suggested that in this organism oxygen may not function in electron transport but rather may act through metalloprotein oxygenases. Tetraethylthiuram disulfide (Antabuse, disulfiram) and its reduction product diethyldithiocarbamate inhibit many metalloprotein oxygenases and have a lipid/H2O partition coefficient and high binding constant for metal ions, favoring selective toxicity to the malaria parasite. These compounds exhibited active antimalarial effects in vitro in concentrations down to 0.1 microgram/ml, the lowest level tested. Tetraethylthiuram disulfide at a level as low as 1 microgram/ml inhibited parasite glycolysis with no effect on glycolysis of normal erythrocytes. Erythrocytes pretreated with this drug at 10 microgram/ml did not support growth of the parasite.


Variation of the dynamic susceptibility along an isochrone.
Koperwas et al. showed in a recent paper [Phys. Rev. Lett. 111, 125701 (2013)] that the dynamic susceptibility χ4 as estimated by dielectric measurements for certain glass-forming liquids decreases substantially with increasing pressure along a curve of constant relaxation time. This observation is at odds with other measures of dynamics being invariant and seems to pose a problem for theories of glass formation. We show that this variation is in fact consistent with predictions for liquids with hidden scale invariance: Measures of dynamics at constant volume are invariant along isochrones, called isomorphs in such liquids, but contributions to fluctuations from long-wavelength fluctuations can vary. This is related to the known noninvariance of the isothermal bulk modulus. Considering the version of χ4 defined for the NVT ensemble, data from simulations of a binary Lennard-Jones liquid show in fact a slight increase with increasing density. This is a true departure from the formal invariance expected for this quantity.


Electron density learning of non-covalent systems.
Chemists continuously harvest the power of non-covalent interactions to control phenomena in both the micro- and macroscopic worlds. From the quantum chemical perspective, the strategies essentially rely upon an in-depth understanding of the physical origin of these interactions, the quantification of their magnitude and their visualization in real-space. The total electron density ρ( r ) represents the simplest yet most comprehensive piece of information available for fully characterizing bonding patterns and non-covalent interactions. The charge density of a molecule can be computed by solving the Schrödinger equation, but this approach becomes rapidly demanding if the electron density has to be evaluated for thousands of different molecules or very large chemical systems, such as peptides and proteins. Here we present a transferable and scalable machine-learning model capable of predicting the total electron density directly from the atomic coordinates. The regression model is used to access qualitative and quantitative insights beyond the underlying ρ( r ) in a diverse ensemble of sidechain-sidechain dimers extracted from the BioFragment database (BFDb). The transferability of the model to more complex chemical systems is demonstrated by predicting and analyzing the electron density of a collection of 8 polypeptides.


Protein kinase Cη polymorphism and the susceptibilities to intracerebral hemorrhage in the Taiwan population.
Protein kinase Cη (PRKCH) 1425G/A polymorphism has been suggested to be a susceptible genetic locus of stroke by genome-wide polymorphism analysis with several replication reports in the Asian populations. While this association was found to exist in the stroke subtype of lacunar infarction especially, the susceptibility of this genetic variant to the intracerebral hemorrhage (ICH) subtypes is unknown to date. This study examined the associations of PRKCH 1425G/A with ICH as well as the stratification of ICH subtypes in the Taiwan population. Genotyping was determined by PCR-based restriction and single strand conformation polymorphism for 381 controls and 303 ICH patients, including 266 deep ICH and 37 lobar ICH patients. Multivariable logistic regression was used to analyze the associations of PRKCH genotypes with stroke subtypes under dominant models. Covariables including age, sex, hypertension, diabetes mellitus, and total cholesterol level were analyzed to delineate the independency of associations. To account for the multiple testing, permutation testing of 1000 replicates was performed for empirical estimates. Distribution of the genotypes of PRKCH 1425G/A was similar while comparing controls (GG: 61%, GA: 33.3%, and AA: 5.7%) with the total ICH group (GG: 57.8%, GA: 36.6%, and AA: 5.6%, p=0.68) and the deep ICH group (GG: 60.2%, GA: 34.2%, and AA: 5.6%, p=0.99). Distribution differ between controls and lobar ICH (GG: 40.5%, GA: 54.1%, and AA: 5.4%, p=0.04). Multivariable logistic regression adjusting for age and sex showed a significant association of PRKCH 1425G/A with lobar ICH risks in a dominant model (OR=2.4, 95% confidence interval (CI) 1.2 to 4.7, p=0.012). When additionally adjusting for hypertension and cholesterol level, this association remained significant (OR=2.4, 95% CI 1.1 to 5.5, p=0.029). There was a borderline association of minor allele A with lobar ICH when compared with controls (OR=1.73, 95% CI 1.01 to 2.9, p=0.039). The variation was not associated with deep ICH. In conclusion, PRKCH 1425G/A variant was not a risk locus for deep ICH phenotype. PRKCH 1425G/A to the susceptibility of lobar ICH shown in this report needs further replication.


Opening of a respiratory intermediate care unit in a general hospital: impact on mortality and other outcomes.
Respiratory intermediate care units (RICUs) are specialized areas aimed at optimizing the cost-benefit ratio of care. No data exist about the impact of opening a RICU on hospital outcomes. We wondered if opening a RICU may improve the outcomes of patients with acute respiratory failure (ARF), acute exacerbation of chronic obstructive pulmonary disease (AECOPD), or community-acquired pneumonia (CAP). We analyzed the discharge abstracts of 2,372 admissions to the RICU and internal medicine units (IMUs) for ARF, AECOPD, and CAP. The IMUs at the Hospital of Trieste comprise emergency and internal wards. In order to investigate the determinants of outcomes, a matched case-control study was performed using clinical records. The in-hospital mortality rate was lower in the RICU vs. IMUs (5.4 vs. 19.1%, p = 0.0001). Statistical differences did not change when comparing the RICU with the emergency and internal wards. After adjusting for potential confounders, the risk of death for patients with CAP, AECOPD, or ARF was significantly higher in the IMUs than in the RICU (OR 6.90, 3.19, and 6.7, respectively, p < 0.04). Both the frequency of transfer to the ICU (6 vs. 12%, p = 0.0001, OR 0.38) and the hospital stay (9.3 vs. 12.1 days, p = 0.0001) were reduced in patients admitted to the RICU compared to those admitted to non-RICUs. Significant differences were found in care management concerning chest physiotherapy, mechanical ventilation, antibiotics, and corticosteroids. The opening of a RICU may be advantageous to reduce in-hospital mortality, the need for ICU admission, and the hospital stay of patients with AECOPD, CAP, and ARF. Better use of care resources contributed to better patient management in the RICU.


Cross-Peaks in Simple Two-Dimensional NMR Experiments from Chemical Exchange of Transverse Magnetisation.
Two-dimensional correlation measurements such as COSY, NOESY, HMQC, and HSQC experiments are central to small-molecule and biomolecular NMR spectroscopy, and commonly form the basis of more complex experiments designed to study chemical exchange occurring during additional mixing periods. However, exchange occurring during chemical shift evolution periods can also influence the appearance of such spectra. While this is often exploited through one-dimensional lineshape analysis ("dynamic NMR"), the analysis of exchange across multiple chemical shift evolution periods has received less attention. Here we report that chemical exchange-induced cross-peaks can arise in even the simplest two-dimensional NMR experiments. These cross-peaks can have highly distorted phases that contain rich information about the underlying exchange process. The quantitative analysis of such peaks, from a single 2D spectrum, can provide a highly accurate characterisation of underlying exchange processes.


Yeast Augmented Network Analysis (YANA): a new systems approach to identify therapeutic targets for human genetic diseases.
Genetic interaction networks that underlie most human diseases are highly complex and poorly defined. Better-defined networks will allow identification of a greater number of therapeutic targets. Here we introduce our Yeast Augmented Network Analysis (YANA) approach and test it with the X-linked spinal muscular atrophy (SMA) disease gene UBA1. First, we express UBA1 and a mutant variant in fission yeast and use high-throughput methods to identify fission yeast genetic modifiers of UBA1. Second, we analyze available protein-protein interaction network databases in both fission yeast and human to construct UBA1 genetic networks. Third, from these networks we identified potential therapeutic targets for SMA. Finally, we validate one of these targets in a vertebrate (zebrafish) SMA model. This study demonstrates the power of combining synthetic and chemical genetics with a simple model system to identify human disease gene networks that can be exploited for treating human diseases.


Evaluation of D-dimer in the diagnosis of suspected deep-vein thrombosis.
Several diagnostic strategies using ultrasound imaging, measurement of D-dimer, and assessment of clinical probability of disease have proved safe in patients with suspected deep-vein thrombosis, but they have not been compared in randomized trials. Outpatients presenting with suspected lower-extremity deep-vein thrombosis were potentially eligible. Using a clinical model, physicians evaluated the patients and categorized them as likely or unlikely to have deep-vein thrombosis. The patients were then randomly assigned to undergo ultrasound imaging alone (control group) or to undergo D-dimer testing (D-dimer group) followed by ultrasound imaging unless the D-dimer test was negative and the patient was considered clinically unlikely to have deep-vein thrombosis, in which case ultrasound imaging was not performed. Five hundred thirty patients were randomly assigned to the control group, and 566 to the D-dimer group. The overall prevalence of deep-vein thrombosis or pulmonary embolism was 15.7 percent. Among patients for whom deep-vein thrombosis had been ruled out by the initial diagnostic strategy, there were two confirmed venous thromboembolic events in the D-dimer group (0.4 percent; 95 percent confidence interval, 0.05 to 1.5 percent) and six events in the control group (1.4 percent; 95 percent confidence interval, 0.5 to 2.9 percent; P=0.16) during three months of follow-up. The use of D-dimer testing resulted in a significant reduction in the use of ultrasonography, from a mean of 1.34 tests per patient in the control group to 0.78 in the D-dimer group (P=0.008). Two hundred eighteen patients (39 percent) in the D-dimer group did not require ultrasound imaging. Deep-vein thrombosis can be ruled out in a patient who is judged clinically unlikely to have deep-vein thrombosis and who has a negative D-dimer test. Ultrasound testing can be safely omitted in such patients.


Prostaglandin E production by human blood monocytes and mouse peritoneal macrophages.
Purified populations of both human peripheral blood monocytes and murine peritoneal macrophages synthesize and release Prostaglandin E in vitro. In contrast, prostaglandin E was detected in neither the supernate fluids from cultures of highly enriched human lymphocytes and granulocytes, nor in nonadherent murine peritoneal cells. Macrophage prostaglandin E production was markedly enhanced by endotoxin, and completely suppressed by indomethacin. All neoplastic monocyte-macrophage cell lines examined elaborated prostaglandin E in vitro, either constitutively or after induction with endotoxin. In contrast, prostaglandin E production could not be detected from either a T- or B-cell lymphoma, whether or not they were treated with endotoxin. These findings thus indicate that the blood monocyte and tissue macrophage represent an important source of prostaglandin E, a function shared by both normal and neoplastic mononuclear phagocytes.


Size dependent structural and polymorphic transitions in ZnO: from nanocluster to bulk.
We report on an extensive survey of (ZnO)N nanostructures ranging from bottom-up generated nanoclusters to top-down nanoparticles cuts from bulk polymorphs. The obtained results enable us to follow the energetic preferences of structure and polymorphism in (ZnO)N systems with N varying between 10-1026. This size range encompasses small nanoclusters with 10s of atoms and nanoparticles with 100s of atoms, which we also compare with appropriate bulk limits. In all cases the nanostructures and bulk systems are optimized using accurate all-electron, relativistic density functional theory based calculations with numeric atom centered orbital basis sets. Specifically, sets of five families of (ZnO)N species are considered: single-layered and multi-layered nanocages, and bulk cut nanoparticles from the sodalite (SOD), body centered tetragonal (BCT), and wurtzite (WZ) ZnO polymorphs. Using suitable fits to interpolate and extrapolate these data allows us to assess the size-dependent energetic stabilities of each family. With increasing size our results indicate a progressive change in energetic stability from single-layered to multi-layered cage-like nanoclusters. For nanoparticles of around 2.6 nm diameter we identify a transitional region where multi-layered cages, SOD, and BCT nanostructures are very similar in energetic stability. This transition size also marks the size regime at which bottom-up nanoclusters give way to top-down bulk-cut nanoparticles. Eventually, a final crossover is found where the most stable WZ-ZnO polymorph begins to energetically dominate at N ∼ 2200. This size corresponds to an approximate nanoparticle diameter of 4.7 nm, in line with experiments reporting the observation of wurtzite crystallinity in isolated ligand-free ZnO nanoparticles of 4-5 nm size or larger.


Differences at surgery between patients with bicuspid and tricuspid aortic valves.
To determine differences in surgical procedures and clinical characteristics at the time of surgery between native bicuspid aortic valves (BAV) and tricuspid aortic valves (TAV) in patients being followed up after aortic valve surgery (AVS). In this retrospective cohort study in a non-academic hospital, we identified patients who had a surgeon's report of the number of native valve cusps and were still being followed up. We selected patients with BAV and TAV, and used multivariable regression analyses to identify associations between BAV-TAV and pre-specified clinical characteristics. Of 439 patients, 140 had BAV (32%) and 299 TAV (68%). BAV patients were younger at the time of surgery (mean age 58.6 ± 13 years) than TAV patients (69.1 ± 12 years, p < 0.001) and were more often male (64% vs 53%; p = 0.029). Cardiovascular risk factors were less prevalent in BAV than in TAV patients at the time of surgery (hypertension (31% vs 55%), hypercholesterolaemia (29% vs 58%) and diabetes (7% vs 16%); all p < 0.005). Concomitant coronary artery bypass grafting (CABG) was performed less often in BAV than in TAV patients (14% vs 39%, p < 0.001), even when adjusted for confounders (adjusted odds ratio (adj.OR) 0.45; 95% CI: 0.25-0.83). In contrast, surgery of the proximal aorta was performed more often (31% vs 11%, respectively, p < 0.001; adj.OR 2.3; 95% CI: 1.3-4.0). Whereas mechanical stress is the supposed major driver of valvulopathy towards AVS in BAV, prevalent cardiovascular risk factors are a suspected driver towards the requirement for AVS and concomitant CABG in TAV, an observation based on surgical determination of the number of valve cusps.


Arginine 66 Controls Dark-State Formation in Green-to-Red Photoconvertible Fluorescent Proteins.
Photoactivated localization microscopy (PALM) is a powerful technique to investigate cellular nanostructures quantitatively and dynamically. However, the use of PALM for molecular counting or single-particle tracking remains limited by the propensity of photoconvertible fluorescent protein markers (PCFPs) to repeatedly enter dark states. By designing the single mutants mEos2-A69T and Dendra2-T69A, we completely swapped the blinking behaviors of mEos2 and Dendra2, two popular PCFPs. We combined X-ray crystallography and single-molecule microscopy to show that blinking in mEos2 and Dendra2 is largely controlled by the orientation of arginine 66, a highly conserved residue in Anthozoan PCFPs. The Arg66 side-chain conformation affects the bleaching and the on-to-off transition quantum yields, as well as the fraction of molecules entering long-lived dark states, resulting in widely different apparent blinking behaviors that largely modulate the efficiency of current blinking correction procedures. The present work provides mechanistic insight into the complex photophysics of Anthozoan PCFPs and will facilitate future engineering of bright and low-blinking variants suitable for PALM.


PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis.
AXL is expressed in many types of cancer and promotes cancer cell survival, metastasis and drug resistance. Here, we focus on identifying modulators that regulate AXL at the mRNA level. We have previously observed that the AXL promoter activity is inversely correlated with the AXL expression levels, suggesting that post-transcriptional mechanisms exist that down-regulate the expression of AXL mRNA. Here we show that the RNA binding protein PTBP1 (polypyrimidine tract-binding protein) directly targets the 5'-UTR of AXL mRNA in vitro and in vivo. Moreover, we also demonstrate that PTBP1, but not PTBP2, inhibits the expression of AXL mRNA and the RNA recognition motif 1 (RRM1) of PTBP1 is crucial for this interaction. To clarify how PTBP1 regulates AXL expression at the mRNA level, we found that, while the transcription rate of AXL was not significantly different, PTBP1 decreased the stability of AXL mRNA. In addition, over-expression of AXL may counteract the PTBP1-mediated apoptosis. Knock-down of PTBP1 expression could enhance tumor growth in animal models. Finally, PTBP1 was found to be negatively correlated with AXL expression in lung tumor tissues in Oncomine datasets and in tissue micro-array (TMA) analysis. In conclusion, we have identified a molecular mechanism of AXL expression regulation by PTBP1 through controlling the AXL mRNA stability. These findings may represent new thoughts alternative to current approaches that directly inhibit AXL signaling and may eventually help to develop novel therapeutics to avoid cancer metastasis and drug resistance.


Relationship of pesticide spraying to signs and symptoms in Indonesian farmers.
This study assessed correlations between exposure to pesticides and signs and symptoms of pesticide toxicity among Indonesian farmers. Detailed observations were recorded of spray frequency and pesticide handling, dermal exposure, and the chemicals used. Symptoms of acute illness were reported by the farmers, and signs of poisoning were observed by the interviewers at the time of spraying or within a few hours after it. The spray practices substantially exposed the farmers to pesticides. Signs and symptoms occurred significantly more often during spraying than during nonspraying seasons. Twenty-one percent of the spray operations resulted in three or more neurobehavioral, respiratory, and intestinal signs or symptoms. The number of spray operations per week, the use of hazardous pesticides, and skin and clothes being wetted with the spray solution were significantly and independently associated with the number of signs and symptoms. A dose-effect relationship was found between the neurobehavioral signs and symptoms and the use of multiple organophosphates. For farmers in the tropics, fully protective garb is too hot and too costly to maintain; farmers thus accept illness as a necessity. Integrated pest management has previously been demonstrated to reduce pesticide use with no loss of crop yield. The frequency of spraying should be reduced through widespread training in integrated pest management, and also the licensing and sale of the most hazardous pesticides should be regulated.


Action recognition from one example.
We present a novel action recognition method based on space-time locally adaptive regression kernels and the matrix cosine similarity measure. The proposed method uses a single example of an action as a query to find similar matches. It does not require prior knowledge about actions, foreground/background segmentation, or any motion estimation or tracking. Our method is based on the computation of novel space-time descriptors from the query video which measure the likeness of a voxel to its surroundings. Salient features are extracted from said descriptors and compared against analogous features from the target video. This comparison is done using a matrix generalization of the cosine similarity measure. The algorithm yields a scalar resemblance volume, with each voxel indicating the likelihood of similarity between the query video and all cubes in the target video. Using nonparametric significance tests by controlling the false discovery rate, we detect the presence and location of actions similar to the query video. High performance is demonstrated on challenging sets of action data containing fast motions, varied contexts, and complicated background. Further experiments on the Weizmann and KTH data sets demonstrate state-of-the-art performance in action categorization.


Reduced HIV-1 long terminal repeat transcription in subjects with protective interferon regulatory factor-1 genotype: a potential mechanism mediating resistance to infection by HIV-1.
We previously described the polymorphism in the interferon regulatory factor-1 (IRF-1) gene as a novel correlate of resistance to HIV-1 infection in a Kenyan female sex worker cohort. However, the underlying mechanisms likely mediating this association remained to be elucidated. The initiation of HIV-1 long terminal repeat (LTR) transcription in peripheral blood mononuclear cells (PBMCs) from subjects with different IRF-1 haplotypes, representing protective, intermediate and the least protective IRF-1 allele combinations, were investigated here. A single-cycle pseudovirus construct expressing vesicular stomatitis virus envelop G-protein (VSV-G) and having an HIV-1 pNL4.3 backbone with luciferase insert was used to infect PBMCs with different IRF-1 haplotypes. The efficiency of early HIV-1 LTR transcription was monitored using a luciferase assay. IRF-1 protein levels induced by the infection were measured by quantitative Western blot. Our results showed that PBMCs with the protective IRF-1 genotype demonstrated significantly lower HIV-1 LTR transcription during the initial stages of infection compared to PBMCs with other haplotypes, which correlated with the kinetics of IRF-1 responsiveness to HIV-1 infection in the cells. It suggests that IRF-1 genotypes alter the efficiency of early HIV-1 LTR transcription, likely via modulating expression of IRF-1. This may represent one mechanism mediating the association between IRF-1 polymorphisms and resistance to HIV-1 infection.


Non-retinovascular leakage in diabetic maculopathy.
Diabetic macular oedema is the leading cause of vision deterioration in diabetic retinopathy. Extracellular fluid within the retina, which distorts the retinal architecture, was assumed to be strictly of retinal vasculature origin. However, there is some experimental evidence supporting clinical observations suggesting a possible role of the retinal pigment epithelium (RPE). An unusual form of diabetic maculopathy is presented in which the RPE and the subretinal space play the main role. Fluorescein angiograms of 1850 non-proliferative diabetic retinopathy (NPDR) patients were examined. Nineteen eyes (14 patients, 1% of NPDR patients) met the criteria-mainly having minimal diabetic retinopathy with only a few microaneurysms and no clinically significant macular oedema (CSMO). Early phase angiograms were compared with late phase angiograms. It was found that in all 19 eyes the area of diffuse RPE late phase leakage was spread around the macular area. No cystic changes or cystoid macular oedema were present in any of the eyes and the visual acuity was 6/10 or better in all the eyes. Possible changes occur in the RPE that may be responsible for the late leakage in NPDR patients-namely, diabetic retinal pigment epitheliopathy. These changes are associated with a breakdown of the outer blood-retinal barrier, consisting of leakage through RPE cells. No focal or diffuse leakage across the RPE has been reported in the literature related to the fluorescein angiograms in diffuse diabetic maculopathy.


Breaking bad news sensitively: what is important to patients in their last year of life?
To understand patients' perspectives on how a diagnosis of a life-limiting illness was first communicated to them. In-depth qualitative interviews with 50 people ranging in age from 30 to 93 years, diagnosed with cancer (31), heart failure (13), stroke (three) or neurological conditions (three) and thought by the responsible health professional to be in the last year of life. Participants from two areas of Northern England were recruited through specialist nurses or hospital consultants and interviewed in their current place of residence. Transcribed data were analysed using Framework. Patients were most likely to recall the pace and clarity with which bad news was conveyed. A direct approach was most common, without much prior warning for the patients. Direct information was usually received well when the patient knew the health professional and when it had been suggested that the patient should be accompanied to the appointment. Some professionals did work to set the scene for the eventual news, with a gradual build-up of information, and narrowing down of options. This approach was perceived as appropriate and sensitive. People with heart failure had engaged in much less discussion about their condition, and most did not recall a specific conversation with their doctor about their prognosis. Bad news is not always broken in a sensitive way, despite considerable efforts to address this issue. Relatively minor changes to practice could improve the patient experience: greater preparation, provision of sufficient time in consultations and cautious disclosure for new patients.


Community integration 2 years after moderate and severe traumatic brain injury.
The aim of this study was to examine community integration by the Community Integration Questionnaire (CIQ) 2 years after injury in a divided TBI sample of moderately and severely injured patients. The second aim was to identify social-demographic, injury-related and rehabilitation associated predictors of CIQ. A cohort study. Outpatient follow-up. Fifty-seven patients with moderate (n = 21) or severe (n = 36) TBI were examined with the Community Integration Questionnaire (CIQ) at 2 years after injury. Possible predictors were analysed in a regression model using CIQ total score at 2 years as the outcome measure. The Community Integration Questionnaire. At 2 years follow-up, there was significant difference between the moderately and severely injured patients in the productivity scores (p < 0.003), while difference in the total CIQ scores approached the significance level (p = 0.074). Significant predictors of a higher CIQ score were living with a spouse, higher Glasgow Coma Scale (GCS) in the acute phase, shorter Post-Traumatic Amnesia (PTA), longer rehabilitation stay (LOS) and use of rehabilitation service. Use of rehabilitation service (B = 7.766) and living with a spouse (B = 4.251) had the largest influence. This means that living with a spouse, better score on the GCS scale, shorter PTA, longer LOS and use of rehabilitation service after discharge equated to better community integration 2 years after TBI Conclusions: Two years after TBI the moderately injured patients have a higher productivity level than the severely injured patients. Marital status, injury severity and rehabilitation after injury were associated with community integration 2 years after TBI.


Controversies in timing of the first dose of anticoagulant prophylaxis against venous thromboembolism after major orthopedic surgery.
Adjusted doses of oral warfarin sodium or fixed doses of subcutaneous low-molecular-weight heparin (LMWH) are the standard approaches for preventing venous thromboembolism following major orthopedic surgery of the legs. In recent years, new anticoagulants have been compared with either LMWH or warfarin. The optimal timing for the first dose of LMWH prophylaxis and of the new anticoagulants is controversial. Recent clinical trials of LMWH and of newer anticoagulants have provided new information on the relationship between the timing of the first anticoagulant dose and the efficacy and safety of thromboprophylaxis after major orthopedic surgery. These data on the optimal timing of initiating prophylaxis come from limited direct randomized comparisons of different timing with the same anticoagulant, subgroup analysis of large studies with a single anticoagulant, indirect comparisons across studies in systematic reviews, and single randomized trials comparing different anticoagulants. In the direct comparison of the same anticoagulant, preoperative initiation of the same regimen of LMWH (dalteparin) increased major bleeding, without improved antithrombotic efficacy compared to the early postoperative regimen. Fondaparinux, 2.5 mg, begun 6 h postoperatively is more effective and as safe as the currently approved regimens of enoxaparin begun either 12 h preoperatively, or 12 to 24 h postoperatively, in patients undergoing major orthopedic surgery. In a subgroup analysis of several large randomized trials, fondaparinux, 2.5 mg, begun < 6 h postoperatively was associated with increased major bleeding, without improved efficacy. The results of indirect comparisons also favor the use of a 6-h postoperative starting time for the first dose, while the single randomized trials comparing different anticoagulants performed to date are not helpful in establishing an optimal time for the first dose. The aggregate clinical research evidence supports the following general conclusions about the relationship between the timing of the first anticoagulant dose and the efficacy and safety of prophylaxis: (1) preoperative initiation is not required for good efficacy and, when begun within 2 h of surgery, increases major bleeding; (2) initiation at 6 h postoperatively is effective and not associated with increased major bleeding; (3) initiation < 6 h postoperatively increases major bleeding, without improved efficacy; thus, 6 h appears to be the threshold for early postoperative administration; and (4) initiation 12 to 24 h postoperatively may be less effective than initiation at 6 h, but further randomized trials comparing the same anticoagulant initiated at different times postoperatively (eg, 6 h vs 12 h) are required to establish definitively the optimal timing of the first anticoagulant dose.


Hydrochlorothiazide improves ventricular compliance and thus performance without reducing hypertrophy in renal artery stenosis in rats.
The effect of hydrochlorothiazide (1 mg/kg per day) on left ventricular (LV) mass and systolic and diastolic function was investigated in two-kidney, one clip (2K1C) renovascular hypertensive rats. Hydrochlorothiazide was administered from 8 weeks, and LV mass and function were measured at 16 weeks after surgery to induce hypertension. Cardiac performance was determined from cardiac output, stroke volume (per 100 g of body weight), and stroke work (per gram of LV weight) versus LV end-diastolic pressure (LVEDP) and versus LV strain relations in anesthetized open-chest, ventilated rats. LV compliance was determined from the LVEDP versus strain relation. Strain was calculated from LV end-diastolic short-axis diameter values. Hydrochlorothiazide reduced systolic blood pressure in 2K1C rats to levels similar to those in sham-operated controls (sham) at 12 weeks after surgery. A reduced afterload failed to influence LV mass, as left LV hypertrophy developed to the same extent in treated 2K1C rats. 2K1C hypertension produced abnormal cardiac performance with altered cardiac output, stroke volume, and stroke work versus LVEDP relations (stroke work versus LVEDP, intercept of 2K1C versus sham, p < 0.001). This was attributed to a decreased ventricular compliance (strain versus LVEDP, slope of 2K1C versus sham, p < 0.001). In contrast, hydrochlorothiazide improved ventricular compliance (strain versus LVEDP, slope of 2K1C versus 2K1C hydrochlorothiazide, p < 0.01) and thus returned the stroke work versus LVEDP relation to sham values (intercept of 2K1C versus 2K1C hydrochlorothiazide, p < 0.001). We conclude that hydrochlorothiazide reduces blood pressure but not the development of ventricular hypertrophy in 2K1C rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of a tomato-rich diet on markers of cardiovascular disease risk in moderately overweight, disease-free, middle-aged adults: a randomized controlled trial.
Cardiovascular disease (CVD) is a major cause of mortality in the United Kingdom. Epidemiologic studies suggest that consumption of tomato-based foods may lower CVD risk. Such potential benefits have been ascribed in part to high concentrations of lycopene in the tomatoes. However, these findings have not yet been validated by comprehensive intervention trials. The aim of this study was to conduct a single-blind, randomized controlled intervention trial with healthy middle-aged volunteers to assess whether the consumption of tomato-based foods affects recognized biomarkers of CVD risk. After a 4-wk run-in period with a low-tomato diet, 225 volunteers (94 men and 131 women) aged 40-65 y were randomly assigned into 1 of 3 dietary intervention groups and asked to consume a control diet (low in tomato-based foods), a high-tomato-based diet, or a control diet supplemented with lycopene capsules (10 mg/d) for 12 wk. Blood samples were collected at baseline, at 6 wk, and after the intervention and were analyzed for carotenoid and lipid profiles and inflammatory markers. Blood pressure, weight, and arterial stiffness were also measured. Dietary intake was also determined during the intervention. None of the systemic markers (inflammatory markers, markers of insulin resistance and sensitivity) changed significantly after the dietary intervention. Moreover, lipid concentrations and arterial stiffness were also unaffected by the interventions. These data indicate that a relatively high daily consumption of tomato-based products (equivalent to 32-50 mg lycopene/d) or lycopene supplements (10 mg/d) is ineffective at reducing conventional CVD risk markers in moderately overweight, healthy, middle-aged individuals. This trial was registered at isrctn.org as ISRCTN34203810.