Testing for BNP and NT-proBNP in the diagnosis and prognosis of heart failure.
The purpose of this systematic review was to evaluate BNP and NT-proBNP to: (a) identify determinants, (b) establish their diagnostic performance in heart failure (HF) patients, (c) determine their predictive ability with respect to mortality and other cardiac endpoints, and (d) determine their value in monitoring HF treatment. MEDLINE, EMBASE, CINAHL, Cochrane Central, and AMED from 1989 to February 2005 were searched for primary studies. Standard systematic review methodology, including meta-analysis, was employed. All study designs were included. Eligibility criteria included English-only studies and restricted the number of test methods to maximize generalizability. Outcomes for prognosis were limited to mortality and specific cardiac events. Further specific criteria were developed for each research question. Determinants: There were 103 determinants identified including age, gender, disease, treatment, as well as biochemical and physiological measures. Few studies reported independent associations and of those that did age, female gender and creatinine levels were positively associated with BNP and NT-proBNP. Pooled sensitivity and specificity values were 94 and 66 percent for BNP and 92 and 65 percent for NT-proBNP; there was minimal difference among settings (emergency, specialized clinics, and primary care). B-type natriuretic peptides also added independent diagnostic information above traditional measures for HF. Both BNP and NT-proBNP were found to be independent predictors of mortality and other cardiac composite endpoints in patients with risk of coronary artery disease (CAD) (risk estimate range = 1.10 to 5.40), diagnosed CAD (risk estimate range = 1.50 to 3.00), and diagnosed HF patients (risk estimate range = 2.11 to 9.35). With respect to screening, the AUC values (range = 0.57 to 0.88) suggested poor performance. Monitoring Treatment: Studies showed therapy reduced BNP and NT-proBNP, however, relationship to outcome was limited and not consistent. Determinants: The importance of the identified determinants for clinical use is not clear. In all settings both BNP and NT-proBNP show good diagnostic properties as a rule out test for HF. BNP and NT-proBNP are consistent independent predictors of mortality and other cardiac composite endpoints for populations with risk of CAD, diagnosed CAD, and diagnosed HF. There is insufficient evidence to determine the value of B-type natriuretic peptides for screening of HF. Monitoring Treatment: There is insufficient evidence to demonstrate that BNP and NT-proBNP levels show change in response to therapies to manage stable chronic HF patients.


Technology and Technique Standards for Camera-Acquired Digital Dermatologic Images: A Systematic Review.
Photographs are invaluable dermatologic diagnostic, management, research, teaching, and documentation tools. Digital Imaging and Communications in Medicine (DICOM) standards exist for many types of digital medical images, but there are no DICOM standards for camera-acquired dermatologic images to date. To identify and describe existing or proposed technology and technique standards for camera-acquired dermatologic images in the scientific literature. Systematic searches of the PubMed, EMBASE, and Cochrane databases were performed in January 2013 using photography and digital imaging, standardization, and medical specialty and medical illustration search terms and augmented by a gray literature search of 14 websites using Google. Two reviewers independently screened titles of 7371 unique publications, followed by 3 sequential full-text reviews, leading to the selection of 49 publications with the most recent (1985-2013) or detailed description of technology or technique standards related to the acquisition or use of images of skin disease (or related conditions). No universally accepted existing technology or technique standards for camera-based digital images in dermatology were identified. Recommendations are summarized for technology imaging standards, including spatial resolution, color resolution, reproduction (magnification) ratios, postacquisition image processing, color calibration, compression, output, archiving and storage, and security during storage and transmission. Recommendations are also summarized for technique imaging standards, including environmental conditions (lighting, background, and camera position), patient pose and standard view sets, and patient consent, privacy, and confidentiality. Proposed standards for specific-use cases in total body photography, teledermatology, and dermoscopy are described. The literature is replete with descriptions of obtaining photographs of skin disease, but universal imaging standards have not been developed, validated, and adopted to date. Dermatologic imaging is evolving without defined standards for camera-acquired images, leading to variable image quality and limited exchangeability. The development and adoption of universal technology and technique standards may first emerge in scenarios when image use is most associated with a defined clinical benefit.


Anti-neutrophil chemokine preserves alveolar development in hyperoxia-exposed newborn rats.
Inflammation may contribute to lung injury and impaired alveolar development in bronchopulmonary dysplasia. We treated hyperoxia-exposed newborn rats with antibodies to the neutrophil chemokine cytokine-induced neutrophil chemoattractant-1 (CINC-1) during 95% O2 exposure to reduce adverse effects of hyperoxia-induced inflammation on lung development. Rats were exposed at birth to air, 95% O2, or 95% O2 + anti-CINC-1 (injected on days 3 and 4). Bromodeoxyuridine (BrdU) was injected 6 h before death. Anti-CINC-1 treatment improved weight gain but not survival at day 8. Anti-CINC-1 reduced bronchoalveolar lavage neutrophils at day 8 to levels equal to air controls. Total detectable lung CINC-1 was reduced to air control levels. Lung compliance was improved by anti-CINC-1, achieving air control levels in the 10-microg anti-CINC-1 group. Anti-CINC-1 preserved proliferating cell nuclear antigen expression in airway epithelium despite 95% O2 exposure. BrdU incorporation was depressed by hyperoxia but preserved by anti-CINC-1 to levels similar to air control. Alveolar volume and surface density were decreased by hyperoxia but preserved by anti-CINC-1 to levels equal to air control. Blockade of neutrophil influx in newborns may avert early lung injury and avoid alveolar developmental arrest that contributes to bronchopulmonary dysplasia.


Cognitive benefits of the ketogenic diet in patients with epilepsy: A systematic overview.
The ketogenic diet (KD) has been found to be effective in reducing seizures in patients with treatment-refractory epilepsy. Less attention has been paid to additional cognitive benefits of KD. The aim of the present paper was to provide a comprehensive overview of the studies reporting effects on cognition after KD treatment in adults and children with epilepsy. To address this aim, the clinical literature on cognitive effects of KD in patients with epilepsy was reviewed using a systematic approach. We conclude that using subjective assessments of the patient's experience, cognitive improvements are frequently reported during KD treatment in the domains of alertness, attention, and global cognition. Studies that used objective neuropsychological tests confirmed benefits on alertness but found no improvement in global cognition. There are indications that these improvements are caused by both seizure reduction and direct effects of KD on cognition. The improvements appear to be unrelated to medication reduction, age when KD is started, type of KD, and sleep improvement. The findings in the present overview contribute to a better understanding of the beneficial effects of KD in patients with epilepsy.


Association of increased intramyocellular lipid content with insulin resistance in lean nondiabetic offspring of type 2 diabetic subjects.
Insulin resistance plays an important role in the pathogenesis of type 2 diabetes; however, the multiple mechanisms causing insulin resistance are not yet fully understood. The aim of this study was to explore the possible contribution of intramyocellular lipid content in the pathogenesis of skeletal muscle insulin resistance. We compared insulin-resistant and insulin-sensitive subjects. To meet stringent matching criteria for other known confounders of insulin resistance, these individuals were selected from an extensively metabolically characterized group of 280 first-degree relatives of type 2 diabetic subjects. Some 13 lean insulin-resistant and 13 lean insulin-sensitive subjects were matched for sex, age, BMI, percent body fat, physical fitness, and waist-to-hip ratio. Insulin sensitivity was determined by the hyperinsulinemic-euglycemic clamp method (for insulin-resistant subjects, glucose metabolic clearance rate [MCR] was 5.77+/-0.28 ml x kg(-1) x min(-1) [mean +/- SE]; for insulin-sensitive subjects, MCR was 10.15+/-0.7 ml x kg(-1) x min(-1); P<0.002). Proton magnetic resonance spectroscopy (MRS) was used to measure intramyocellular lipid content (IMCL) in both groups. MRS studies demonstrated that in soleus muscle, IMCL was increased by 84% (11.8+/-1.6 vs. 6.4+/-0.59 arbitrary units; P = 0.008 ), and in tibialis anterior muscle, IMCL was increased by 57% (3.26+/-0.36 vs. 2.08+/-0.3 arbitrary units; P = 0.017) in the insulin-resistant offspring, whereas the extramyocellular lipid content and total muscle lipid content were not statistically different between the two groups. These data demonstrate that in these well-matched groups of lean subjects, IMCL is increased in insulin-resistant offspring of type 2 diabetic subjects when compared with an insulin-sensitive group matched for age, BMI, body fat distribution, percent body fat, and degree of physical fitness. These results indicate that increased IMCL represents an early abnormality in the pathogenesis of insulin resistance and suggest that increased IMCL may contribute to the defective glucose uptake in skeletal muscle in insulin-resistant subjects.


European lactase persistence genotype shows evidence of association with increase in body mass index.
The global prevalence of obesity has increased significantly in recent decades, mainly due to excess calorie intake and increasingly sedentary lifestyle. Here, we test the association between obesity measured by body mass index (BMI) and one of the best-known genetic variants showing strong selective pressure: the functional variant in the cis-regulatory element of the lactase gene. We tested this variant since it is presumed to provide nutritional advantage in specific physical and cultural environments. We genetically defined lactase persistence (LP) in 31 720 individuals from eight European population-based studies and one family study by genotyping or imputing the European LP variant (rs4988235). We performed a meta-analysis by pooling the beta-coefficient estimates of the relationship between rs4988235 and BMI from the nine studies and found that the carriers of the allele responsible for LP among Europeans showed higher BMI (P = 7.9 x 10(-5)). Since this locus has been shown to be prone to population stratification, we paid special attention to reveal any population substructure which might be responsible for the association signal. The best evidence of exclusion of stratification came from the Dutch family sample which is robust for stratification. In this study, we highlight issues in model selection in the genome-wide association studies and problems in imputation of these special genomic regions.


Abandoned metal mines and their impact on receiving waters: A case study from Southwest England.
Historic mine sites are a major source of contamination to terrestrial and river environments. To demonstrate the importance of determining the significance of point and diffuse metal contamination and the related bioavailability of the metals present from abandoned mines a case study has been carried out. The study provides a quantitative assessment of a historic mine site, Wheal Betsy, southwest England, and its contribution to non-compliance with Water Framework Directive (WFD) Environmental Quality Standards (EQS) for Cd, Cu, Pb and Zn. Surface water and sediment samples showed significant negative environmental impacts even taking account of the bioavailability of the metal present, with lead concentration in the stream sediment up to 76 times higher than the Canadian sediment guidelines 'Probable Effect Level'. Benthic invertebrates showed a decline in species richness adjacent to the mine site with lead and cadmium the main cause. The main mine drainage adit was the single most significant source of metal (typically 50% of metal load from the area, but 88% for Ni) but the mine spoil tips north and south of the adit input added together discharged roughly an equivalent loading of metal with the exception of Ni. The bioavailability of metal in the spoil tips exhibited differing spatial patterns owing to varying ambient soil physico-chemistry. The data collected is essential to provide a clear understanding of the contamination present as well as its mobility and bioavailability, in order to direct the decision making process regarding remediation options and their likely effectiveness.


Addressing behavioral impacts of childhood leukemia: A feasibility pilot randomized controlled trial of a group videoconferencing parenting intervention.
Child emotional and behavioral problems constitute significant sequelae of acute lymphoblastic leukemia (ALL) treatment. The aims of this study were to a) examine the feasibility, acceptability and satisfaction of a parenting intervention amongst parents of children with ALL and b) explore whether participation in a parenting intervention shows promise for improvements in child behavior. 12 parents with a child aged between 2 and 8 years receiving maintenance phase treatment for ALL participated in a phase 2 randomized controlled trial comparing eight weeks of group online participation in Triple P: Positive Parenting Program with no intervention. The number of eligible parents who completed the intervention was low (31.6%). Main reasons for non-consent or dropout were program time commitment too high or content not relevant. For parents who completed the intervention, satisfaction and acceptability was high. Parents reported the intervention as highly relevant and topical, feasible, helpful and a positive experience. Results indicated a non-significant trend towards improved total child behavioral and emotional difficulties following the intervention. Qualitative results indicated that intervention group parents reported improvements in parenting skills and competence, and decreased child behavioral problems. These pilot data highlight the difficulties of engaging and retaining parents in an 8-week parenting intervention in this context. For parents who completed the intervention, results indicated high feasibility, acceptability and satisfaction. Suggestions for further research and intervention modifications are provided to enhance uptake and strengthen efforts to assist parents in addressing child behavioral and emotional challenges during ALL treatment.


Novel mechanisms of tube-size regulation revealed by the Drosophila trachea.
The size of various tubes within tubular organs such as the lung, vascular system and kidney must be finely tuned for the optimal delivery of gases, nutrients, waste and cells within the entire organism. Aberrant tube sizes lead to devastating human illnesses, such as polycystic kidney disease, fibrocystic breast disease, pancreatic cystic neoplasm and thyroid nodules. However, the underlying mechanisms that are responsible for tube-size regulation have yet to be fully understood. Therefore, no effective treatments are available for disorders caused by tube-size defects. Recently, the Drosophila tracheal system has emerged as an excellent in vivo model to explore the fundamental mechanisms of tube-size regulation. Here, we discuss the role of the apical luminal matrix, cell polarity and signaling pathways in regulating tube size in Drosophila trachea. Previous studies of the Drosophila tracheal system have provided general insights into epithelial tube morphogenesis. Mechanisms that regulate tube size in Drosophila trachea could be well conserved in mammalian tubular organs. This knowledge should greatly aid our understanding of tubular organogenesis in vertebrates and potentially lead to new avenues for the treatment of human disease caused by tube-size defects.


ALDH1A1 is a marker for malignant prostate stem cells and predictor of prostate cancer patients' outcome.
Prostate cancer (PCa) contains a small population of cancer stem cells (CSCs) that contribute to its initiation and progression. The development of specific markers for identification of the CSCs may lead to new diagnostic strategies of PCa. Increased aldehyde dehydrogenase 1A1 (ALDH1A1) activity has been found in the stem cell populations of leukemia and some solid tumors. The aim of the study was to investigate the stem-cell-related function and clinical significance of the ALDH1A1 in human PCa. ALDEFLUOR assay was used to isolate ALDH1A1(+) cells from PCa cell lines. Stem cell characteristics of the ALDH1A1(+) cells were then investigated by in vitro and in vivo approaches. The ALDH1A1 expression was also analyzed by immunohistochemistry in 18 normal prostate and 163 PCa tissues. The ALDH1A1(+) PCa cells showed high clonogenic and tumorigenic capacities, and serially reinitiated transplantable tumors that resembled histopathologic characteristics and heterogeneity of the parental PCa cells in mice. Immunohistochemical analysis of human prostate tissues showed that ALDH1A1(+) cells were sparse and limited to the basal component in normal prostates. However, in tumor specimens, increased ALDH1A1 immunopositivity was found not only in secretory type cancer epithelial cells but also in neuroendocrine tumor populations. Furthermore, the high ALDH1A1 expression in PCa was positively correlated with Gleason score (P=0.01) and pathologic stage (P=0.01), and inversely associated with overall survival and cancer-specific survival of the patients (P=0.00093 and 0.00017, respectively). ALDH1A1 could be a prostate CSC-related marker. Measuring its expression might provide a potential approach to study tumorigenesis of PCa and predict outcome of the disease.


The role of the 6 lysines and the terminal amine of Escherichia coli single-strand binding protein in its binding of single-stranded DNA.
Differential chemical modification of the lysines and amino-terminus of Escherichia coli single-strand binding (SSB) protein was used to determine their roles in the binding of SSB to single-stranded DNA (ssDNA). A combination of isotope labeling and mass spectrometry was used to determine the rates at which SSB was acetylated by acetic anhydride. First, SSB was labeled by deuterated acetic anhydride for given lengths of time in the presence or absence of single-stranded ssDNA. Then, the protein was denatured and completely acetylated by nondeuterated acetic anhydride. Enzymatic digests of the completely acetylated, isotopically labeled SSB were analyzed by electrospray ionization mass spectrometry. The intensities of the deuterated and nondeuterated forms of acetylated peptides provided accurate quantification of the reactivity of the amines in native SSB, either free or bound to ssDNA. Acetylation rate constants were determined from time course measurements. In the absence of ssDNA, the terminal alpha-amine of SSB was 10-fold more reactive than Lys residues at positions 43, 62, 73, and 87. The reactivities of Lys 7 and 49 were much lower yet, suggesting that they have very limited access to solution under any condition. In the presence of ssDNA, the reactivities of the amino-terminus and Lys residues 43, 62, 73, and 87 were reduced by factors of 3.7-25, indicating that the environments around all of these amines is substantially altered by binding of SSB to ssDNA. Three of these residues are located near putative ssDNA binding sites, whereas Lys 87 is located at the monomer-monomer interface.


Molecular determination of point mutation haplotypes in the dihydrofolate reductase and dihydropteroate synthase of Plasmodium falciparum in three districts of northern Tanzania.
The antimalarial combination of sulfadoxine and pyrimethamine (SP) was introduced as first-line treatment for uncomplicated malaria in Tanzania during 2001 following 18 years of second-line use. The genetic determinants of in vitro resistance to the two drugs individually are shown to be point mutations at seven sites in the dihydrofolate reductase gene (dhfr) conferring resistance to pyrimethamine and five sites in the dihydropteroate synthase (dhps) gene conferring resistance to sulfadoxine. Different combinations of mutations within each gene confer differing degrees of insensitivity, but information about the frequency with which allelic haplotypes occur has been lacking because of the complicating effects of multiple infection. Here we used a novel high-throughput sequence-specific oligonucleotide probe-based approach to examine the present resistance status of three Plasmodium falciparum populations in northern Tanzania. By using surveys of asymptomatic infections and screening for the presence of all known point mutations in dhfr and dhps genes, we showed that just five dhfr and three dhps allelic haplotypes are present. High frequencies of both triple-mutant dhfr and double-mutant dhps mutant alleles were found in addition to significant interregional heterogeneity in allele frequency. In vivo studies have shown that the cooccurrence of three dhfr mutations and two dhps mutations in an infection prior to treatment is statistically predictive of treatment failure. We have combined data for both loci to determine the frequency of two-locus genotypes. The triple-dhfr/double-dhps genotype is present in all three regions with frequencies ranging between 30 and 63%, indicating that treatment failure rates are likely to be high.


Possible hormonal interaction for eliciting courtship behavior in the male newt, Cynops pyrrhogaster.
Reproductive behavior in amphibians, as in other vertebrate animals, is under the control of multiple hormonal substances. Prolactin (PRL), arginine vasotocin (AVT), androgen, and 7α-hydroxypregnenolone (7α-OH PREG), four such substances with hormonal activity, are known to be involved in the expression of the tail vibration behavior which is the initial step of courtship performed by the male newt, Cynops pyrrhogaster. As current information on the interaction(s) between these hormones in terms of eliciting tail vibration behavior is limited, we have investigated whether the decline of expression of tail vibration behavior due to suppression of the activity of any one of these hormones can be restored by supplying any one of the other three hormones exogenously. Expression of the behavior was determined in terms of incidence (% of test animals exhibiting the behavior) and frequency (number of times that the behavior was repeated during the test period). Neither PRL nor androgen restored the decline in the incidence and frequency of the tail vibration behavior caused by the suppression of the activity of any one of other three hormones. AVT completely restored both the anti-PRL antibody-induced and flutamide (an androgen receptor antagonist)-induced, but not ketoconazole (an inhibitor of the steroidogenic CYP enzymes)-induced decline in the incidence and frequency of the tail vibration behavior. The neurosteroid, 7α-OH PREG, failed to restore flutamide-induced decline in the incidence and frequency of the behavior. However, it was able to restore both anti-PRL antibody-induced and AVT receptor antagonist-induced decline in the incidence, but not in the frequency of the behavior. In another experiment designed to see the activity of hormones enhancing the frequency of the tail vibration behavior, AVT was revealed to be more potent than 7α-OH PREG. The role of each hormonal substance in determining the expression of the tail vibration behavior was discussed based on the results.


Relationships of Sleep Duration, Midpoint, and Variability with Physical Activity in the HCHS/SOL Sueño Ancillary Study.
Short and long sleep duration, later sleep midpoint, and greater intra-individual sleep variability are associated with lower physical activity, but previous research lacks objective and concurrent assessment of sleep and physical activity. This cross-sectional study examined whether sleep duration, midpoint, and variability in duration and midpoint were related to wrist actigraphy-measured physical activity. Participants were 2156 Hispanics/Latinos in the Hispanic Community Health Study/Study of Latinos (HCHS/SOL) Sueño Ancillary Study. Participants wore Actiwatch devices to measure sleep and physical activity via the wrist for ≥5 days. Physical activity was defined as minutes/day in the upper quartile of the sampling distribution's non-sleep activity, capturing light to vigorous physical activity. An inverse linear relationship between sleep duration and physical activity was found such that each additional sleep hour related to 29 fewer minutes of physical activity (B = -28.7, SE = 3.8), p < .01). Variability in sleep midpoint was also associated with physical activity; with each 1-hr increase in variability there were 24 more minutes of physical activity (B = 24.2, SE = 5.6, p < .01). In contrast, sleep midpoint and variability in duration were not associated with physical activity. Sensitivity analyses identified an association of short sleep duration and greater variability in sleep duration with greater accelerometry-derived moderate-to-vigorous physical activity measured at the HCHS/SOL baseline (M = 2.1 years before the sleep assessment). Findings help clarify inconsistent prior research associating short sleep duration and sleep variability with greater health risks but also contribute novel information with simultaneous objective assessments.


Oligoclonal protein bands and Ig isotype switching in multiple myeloma treated with high-dose therapy and hematopoietic cell transplantation.
Multiple myeloma (MM) is usually characterized by production of a single serum monoclonal protein of constant isotype and light-chain restriction. Multiple Ig isotypes and isotype switches, which are rare in untreated patients, are reported to be more common in patients undergoing myeloablative therapy. These additional protein bands, detected by immunofixation electrophoresis (IFE), could be due to altered paraprotein production by the malignant plasma cell clone or oligoclonal Ig production during recovery of B-cell function after myeloablative therapy. We analyzed abnormal protein bands (APB), distinct from the presenting paraprotein, in 550 patients receiving high-dose therapy with autologous hematopoietic cell transplantation at a single institution. Fifty-five patients (10%) had APB, 48 had oligoclonal bands (OB), and 23 had an apparent isotype switch (IS) on IFE (16 had both OB and IS). Morphologic and flow cytometric examination of bone marrow in 17 patients with IS showed no evidence of a clonal plasma cell isotype switch. Patients with APB had significantly higher complete response to therapy (67% v 37%, P = .001). To assess the independent prognostic relevance of APB, a multivariate analysis was performed among 471 patients surviving at least 12 months from first transplant (all patients developing APB had done so by 12 months from first transplant). APB (in 50 patients) was a favorable feature for both event-free (rank 3, P = .004) and overall survival (rank 3, P = .0005). We propose that OB and IS are likely to be due to recovery of Ig production rather than alterations in the biology of the malignant plasma cell clone.


Copper-dependent cytotoxicity of 8-hydroxyquinoline derivatives correlates with their hydrophobicity and does not require caspase activation.
This study reports the structure-activity relationship of a series of 8-hydroxoquinoline derivatives (8-HQs) and focuses on the cytotoxic activity of 5-Cl-7-I-8-HQ (clioquinol, CQ) copper complex (Cu(CQ)). 8-HQs alone cause a dose-dependent loss of viability of the human tumor HeLa and PC3 cells, but the coadministration of copper increases the ligands effects, with extensive cell death occurring in both cell lines. Cytotoxic doses of Cu(CQ) promote intracellular copper accumulation and massive endoplasmic reticulum vacuolization that precede a nonapoptotic (paraptotic) cell death. The cytotoxic effect of Cu(CQ) is reproduced in normal human endothelial cells (HUVEC) at concentrations double those effective in tumor cells, pointing to a potential therapeutic window for Cu(CQ). Finally, the results show that the paraptotic cell death induced by Cu(CQ) does not require nor involve caspases, giving an indication for the current clinical assessment of clioquinol as an antineoplastic agent.


Fluorescence imaging of single Kinesin motors on immobilized microtubules.
Recent developments in optical microscopy and nanometer tracking have greatly improved our understanding of cytoskeletal motor proteins. Using fluorescence microscopy, dynamic interactions are now routinely observed in vitro on the level of single molecules mainly using a geometry, where fluorescently labeled motors move on surface-immobilized filaments. In this chapter, we review recent methods related to single-molecule kinesin motility assays. In particular, we aim to provide practical advice on: how to set up the assays, how to acquire high-precision data from fluorescently labeled kinesin motors and attached quantum dots, and how to analyze data by nanometer tracking.


Early adolescent pregnancy increases risk of incident HIV infection in the Eastern Cape, South Africa: a longitudinal study.
Adolescents having unprotected heterosexual intercourse are at risk of HIV infection and unwanted pregnancy. However, there is little evidence to indicate whether pregnancy in early adolescence increases the risk of subsequent HIV infection. In this paper, we tested the hypothesis that adolescent pregnancy (aged 15 or younger) increases the risk of incident HIV infection in young South African women. We assessed 1099 HIV-negative women, aged 15-26 years, who were volunteer participants in a cluster-randomized, controlled HIV prevention trial in the predominantly rural Eastern Cape province of South Africa. All of these young women had at least one additional HIV test over two years of follow-up. Outcomes were HIV incidence rates per 100 person years and HIV incidence rate ratios (IRRs) estimated by Poisson multivariate models. Three pregnancy categories were created for the Poisson model: early adolescent pregnancy (a first pregnancy at age 15 years or younger); later adolescent pregnancy (a first pregnancy at age 16 to 19 years); and women who did not report an adolescent pregnancy. Models were adjusted for study design, age, education, time since first sexual experience, socio-economic status, childhood trauma and herpes simplex virus type 2 infection. HIV incidence rates were 6.0 per 100 person years over two years of follow-up. The adjusted IRR was 3.02 (95% CI 1.50-6.09) for a pregnancy occurring at age 15 or younger. Women with pregnancies occurring between 16 and 19 years of age did not have a higher incidence of HIV (IRR 1.08; 95% CI 0.64-1.84). Early adolescent pregnancies were associated with higher partner numbers and a greater age difference with partners. Early adolescent pregnancies increase the incidence of HIV among South African women. The higher risk is associated with sexual risk behaviours such as higher partner numbers and a greater age difference with partners rather than a biological explanation of hormonal changes during pregnancy.


Effects of antenatal glucocorticoids on cerebral substrate metabolism in the preterm ovine fetus.
Although the benefits of antenatal glucocorticoids are well known for infants who are born preterm, there is increasing evidence of adverse effects on brain development, which may relate to altered metabolic activity. We have determined the effect of maternal glucocorticoid administration at doses that are used clinically on cerebral substrate metabolism in the preterm ovine fetus. Chronically instrumented pregnant sheep at 0.85 gestation received 2 intramuscular injections of betamethasone at 170 microg/kg maternal weight (n = 13) or saline (n = 10) 24 hours apart together with a continuous infusion of L-[1-(13)C] leucine to the fetus. Fetal cerebral substrate arteriovenous differences (O2, glucose, leucine, leucine enrichment) and blood flow (fluorescent microspheres) were measured at baseline, 24 hours after the first betamethasone/saline injection (late beta/saline 1), and 4 hours after the second betamethasone/saline injection (early beta/saline 2) to obtain substrate deliveries and fractional extractions. Fetal pH, blood gases, and metabolites were little changed in either group over the course of the study, except for glucose values in the betamethasone animals, which increased 1.4- and 1.9-fold, measured late beta 1 and early beta 2, respectively (both P < .01). Cerebral blood flow, although little changed in the control group or at late beta 1, was decreased at early beta 2 by approximately 30% (P < .05). As such, early beta 2 animals showed a decrease in cerebral O2 delivery of approximately 20% (P = .06) and conversely an increase in cerebral glucose delivery of 1.4- and 1.3-fold at late beta 1 (P < .05) and early beta 2 (P = .08), respectively. Fractional extraction values for these substrates were not changed significantly, which resulted in corresponding decreases in estimated O2 uptake and increases in estimated glucose uptake, such that the glucose/oxygen quotient (as an index of glucose oxidative metabolism) measured 1.6 at early beta 2, which was considerably greater than baseline values at 1.1 (P < .05). Fractional extraction values for leucine and leucine enrichment averaged 2%-3%; although somewhat higher in the betamethasone animals, none of the between or within group differences were significant. Fetal cerebral metabolism in the preterm ovine fetus is altered by antenatal glucocorticoid administration, which is comparable with that used in human pregnancy, and includes an acute decrease in cerebral blood flow and a probable increase in anaerobic glucose metabolism. Although likely of short duration in conjunction with peak glucocorticoid levels, these metabolic effects may place the developing brain at added risk for superimposed hypoxic injury.


Probing high-momentum protons and neutrons in neutron-rich nuclei.
The atomic nucleus is one of the densest and most complex quantum-mechanical systems in nature. Nuclei account for nearly all the mass of the visible Universe. The properties of individual nucleons (protons and neutrons) in nuclei can be probed by scattering a high-energy particle from the nucleus and detecting this particle after it scatters, often also detecting an additional knocked-out proton. Analysis of electron- and proton-scattering experiments suggests that some nucleons in nuclei form close-proximity neutron-proton pairs1-12 with high nucleon momentum, greater than the nuclear Fermi momentum. However, how excess neutrons in neutron-rich nuclei form such close-proximity pairs remains unclear. Here we measure protons and, for the first time, neutrons knocked out of medium-to-heavy nuclei by high-energy electrons and show that the fraction of high-momentum protons increases markedly with the neutron excess in the nucleus, whereas the fraction of high-momentum neutrons decreases slightly. This effect is surprising because in the classical nuclear shell model, protons and neutrons obey Fermi statistics, have little correlation and mostly fill independent energy shells. These high-momentum nucleons in neutron-rich nuclei are important for understanding nuclear parton distribution functions (the partial momentum distribution of the constituents of the nucleon) and changes in the quark distributions of nucleons bound in nuclei (the EMC effect)1,13,14. They are also relevant for the interpretation of neutrino-oscillation measurements15 and understanding of neutron-rich systems such as neutron stars3,16.


All-in-One Beaker Method for Large-Scale Production of Metal Oxide Hollow Nanospheres Using Nanoscale Kirkendall Diffusion.
A simple and easily scalable process for the formation of metal oxide hollow nanospheres using nanoscale Kirkendall diffusion called the "all-in-one beaker method" is introduced. The Fe2O3, SnO2, NiO, and Co3O4 hollow nanospheres are successfully prepared by the all-in-one beaker method. The detailed formation mechanism of aggregate-free hematite hollow nanospheres is studied. Dimethylformamide solution containing Fe acetate, polyacrylonitrile (PAN), and polystyrene (PS) transforms into aggregate-free Fe2O3 hollow nanospheres. The porous structure formed by the combustion of PS provides a good pathway for the reducing gas. The carbon matrix formed from PAN acts as a barrier, which can prevent the aggregation of metallic Fe nanopowders by surrounding each particle. The Fe-C bulk material formed as an intermediate product transforms into aggregate-free Fe2O3 hollow nanospheres by the nanoscale Kirkendall diffusion process. The mean size and shell thickness of the hollow Fe2O3 nanospheres measured from the TEM images are 52 and 9 nm, respectively. The discharge capacities of the Fe2O3 nanopowders with hollow and dense structures and the bulk material for the 200th cycle at a current density of 0.5 A g(-1) are 1012, 498, and 637 mA h g(-1), respectively, and their capacity retentions calculated compared to those in the second cycles are 92, 45, and 59%, respectively. Additionally, Fe2O3 hollow nanospheres cycled at 1 A g(-1) after 1000 cycles showed a high discharge capacity of 871 mA h g(-1) (capacity retention was 80% from the second cycle). The Fe2O3, SnO2, NiO, and Co3O4 hollow nanospheres show excellent cycling performances for lithium-ion storage because they have a high contact area with the liquid electrolyte and space for accommodating a huge volume change during cycling.


Strategies to Prevent Readmission in High-Risk Patients with Diabetes: the Importance of an Interdisciplinary Approach.
Patients with diabetes are known to have higher 30-day readmission rates compared to the general inpatient population. A number of strategies have been shown to be effective in lowering readmission rates. A review of the current literature revealed several strategies that have been associated with a decreased risk of readmission in high-risk patients with diabetes. These strategies include inpatient diabetes survival skills education and medication reconciliation prior to discharge to send the patient home with the "right" medications. Other key strategies include scheduling a follow-up phone call soon after discharge and an office visit to adjust the diabetes regimen. The authors identified the most successful strategies to reduce readmissions as well as some institutional barriers to following a transitional care program. Recent studies have identified risk factors in the diabetes population that are associated with an increased risk of readmission as well as interventions to lower this risk. A standardized transitional care program that focuses on providing interventions while reducing barriers to implementation can contribute to a decreased risk of readmission.


Chitosan disrupts the barrier properties of the outer membrane of gram-negative bacteria.
The mode of antimicrobial action of chitosan (polymeric beta-1,4-N-acetylglucosamine) on gram-negative bacteria was studied with special emphasis on its ability to bind to and weaken the barrier function of the outer membrane (OM). Chitosan (250 ppm) at pH 5.3 induced significant uptake of the hydrophobic probe 1-N-phenylnaphthylamine (NPN) in Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium. The effect was reduced (E. coli, salmonellae) or abolished (P. aeruginosa) by MgCl2. No NPN uptake was observed during exposure of the salmonellae to chitosan at pH 7.2. Chitosan also sensitized P. aeruginosa and the salmonellae to the lytic effect of sodium dodecyl sulfate (SDS); such sensitization was not blocked by MgCl2 and was reversible by washing chitosan-treated cells prior to SDS exposure. Chemical and electrophoretic analyses of cell-free supernatants of chitosan-treated cell suspensions showed that interaction of chitosan with E. coli and the salmonellae involved no release of lipopolysaccharide (LPS) or other membrane lipids. However, chitosan rendered E. coli more sensitive to the inhibitory action of dyes and bile acids used in selective media. Highly cationic mutants of S. typhimurium were more resistant to chitosan than the parent strains. Electron microscopy showed that chitosan caused extensive cell surface alterations and covered the OM with vesicular structures. Chitosan thus appeared to bind to the outer membrane, explaining the loss of the barrier function. This property makes chitosan a potentially useful indirect antimicrobial for food protection.


Making room for manoeuvre: addressing gender norms to strengthen the enabling environment for agricultural innovation.
Local gender norms constitute a critical component of the enabling (or disabling) environment for improved agricultural livelihoods - alongside policies, markets, and other institutional dimensions. Yet, they have been largely ignored in agricultural research for development. This viewpoint is based on many years of experience, including a recent major comparative research initiative, GENNOVATE, on how gender norms and agency interact to shape agricultural change at local levels. The evidence suggests that approaches which engage with normative dimensions of agricultural development and challenge underlying structures of inequality, are required to generate lasting gender-equitable development in agriculture and natural resource management.


Volcanogenic fluvial-lacustrine environments in iceland and their utility for identifying past habitability on Mars.
The search for once-habitable locations on Mars is increasingly focused on environments dominated by fluvial and lacustrine processes, such as those investigated by the Mars Science Laboratory Curiosity rover. The availability of liquid water coupled with the potential longevity of such systems renders these localities prime targets for the future exploration of Martian biosignatures. Fluvial-lacustrine environments associated with basaltic volcanism are highly relevant to Mars, but their terrestrial counterparts have been largely overlooked as a field analogue. Such environments are common in Iceland, where basaltic volcanism interacts with glacial ice and surface snow to produce large volumes of meltwater within an otherwise cold and dry environment. This meltwater can be stored to create subglacial, englacial, and proglacial lakes, or be released as catastrophic floods and proglacial fluvial systems. Sedimentary deposits produced by the resulting fluvial-lacustrine activity are extensive, with lithologies dominated by basaltic minerals, low-temperature alteration assemblages (e.g., smectite clays, calcite), and amorphous, poorly crystalline phases (basaltic glass, palagonite, nanophase iron oxides). This paper reviews examples of these environments, including their sedimentary deposits and microbiology, within the context of utilising these localities for future Mars analogue studies and instrument testing.


Substantial Neuroprotective and Neurite Outgrowth-Promoting Activities by Bis(propyl)-cognitin via the Activation of Alpha7-nAChR, a Promising Anti-Alzheimer's Dimer.
The cause of Alzheimer's disease (AD) could be ascribed to the progressive loss of functional neurons in the brain, and hence, agents with neuroprotection and neurite outgrowth-promoting activities that allow for the replacement of lost neurons may have significant therapeutic value. In the current study, the neuroprotective and the neurite outgrowth-promoting activities and molecular mechanisms of bis(propyl)-cognitin (B3C), a multifunctional anti-AD dimer, were investigated. Briefly, B3C (24 h pretreatment) fully protected against glutamate-induced neuronal death in primary cerebellar granule neurons with an IC50 value of 0.08 μM. The neuroprotection of B3C could be abrogated by methyllycaconitine, a specific antagonist of alpha7-nicotinic acetylcholine receptor (α7-nAChR). In addition, B3C significantly promoted neurite outgrowth in both PC12 cells and primary cortical neurons, as evidenced by the increase in the percentage of cells with extended neurites as well as the up-regulation of neuronal markers growth-associated protein-43 and β-III-tubulin. Furthermore, B3C rapidly upregulated the phosphorylation of extracellular signal-regulated kinase (ERK), a critical signaling molecule in neurite outgrowth that is downstream of the α7-nAChR signal pathway. Specific inhibitors of ERK and α7-nAChR, but not those of p38 mitogen-activated protein kinase and c-Jun NH(2)-terminal kinase, blocked the neurite outgrowth as well as ERK activation in PC12 cells induced by B3C. Most importantly, genetic depletion of α7-nAChR significantly abolished B3C-induced neurite outgrowth in PC12 cells. Taken together, our results suggest that B3C provided neuroprotection and neurite outgrowth-promoting activities through the activation of α7-nAChR, which offers a novel molecular insight into the potential application of B3C in AD treatment.


Application of ordinal logistic regression analysis in determining risk factors of child malnutrition in Bangladesh.
The study attempts to develop an ordinal logistic regression (OLR) model to identify the determinants of child malnutrition instead of developing traditional binary logistic regression (BLR) model using the data of Bangladesh Demographic and Health Survey 2004. Based on weight-for-age anthropometric index (Z-score) child nutrition status is categorized into three groups-severely undernourished (< -3.0), moderately undernourished (-3.0 to -2.01) and nourished (≥-2.0). Since nutrition status is ordinal, an OLR model-proportional odds model (POM) can be developed instead of two separate BLR models to find predictors of both malnutrition and severe malnutrition if the proportional odds assumption satisfies. The assumption is satisfied with low p-value (0.144) due to violation of the assumption for one co-variate. So partial proportional odds model (PPOM) and two BLR models have also been developed to check the applicability of the OLR model. Graphical test has also been adopted for checking the proportional odds assumption. All the models determine that age of child, birth interval, mothers' education, maternal nutrition, household wealth status, child feeding index, and incidence of fever, ARI & diarrhoea were the significant predictors of child malnutrition; however, results of PPOM were more precise than those of other models. These findings clearly justify that OLR models (POM and PPOM) are appropriate to find predictors of malnutrition instead of BLR models.


Arabidopsis bHLH100 and bHLH101 control iron homeostasis via a FIT-independent pathway.
Iron deficiency induces a complex set of responses in plants, including developmental and physiological changes, to increase iron uptake from soil. In Arabidopsis, many transporters involved in the absorption and distribution of iron have been identified over the past decade. However, little is known about the signaling pathways and networks driving the various responses to low iron. Only the basic helix-loop-helix (bHLH) transcription factor FIT has been shown to control the expression of the root iron uptake machinery genes FRO2 and IRT1. Here, we characterize the biological role of two other iron-regulated transcription factors, bHLH100 and bHLH101, in iron homeostasis. First direct transcriptional targets of FIT were determined in vivo. We show that bHLH100 and bHLH101 do not regulate FIT target genes, suggesting that they play a non-redundant role with the two closely related bHLH factors bHLH038 and bHLH039 that have been suggested to act in concert with FIT. bHLH100 and bHLH101 play a crucial role in iron-deficiency responses, as attested by their severe growth defects and iron homeostasis related phenotypes on low-iron media. To gain further insight into the biological role of bHLH100 and bHLH101, we performed microarray analysis using the corresponding double mutant and showed that bHLH100 and bHLH101 likely regulate genes involved in the distribution of iron within the plant. Altogether, this work establishes bHLH100 and bHLH101 as key regulators of iron-deficiency responses independent of the master regulator FIT and sheds light on new regulatory networks important for proper growth and development under low iron conditions.


Communication: a minimal model for the diffusion-relaxation backbone dynamics of proteins.
We present a model for the local diffusion-relaxation dynamics of the C(α)-atoms in proteins describing both the diffusive short-time dynamics and the asymptotic long-time relaxation of the position autocorrelation functions. The relaxation rate spectra of the latter are represented by shifted gamma distributions, where the standard gamma distribution describes anomalous slow relaxation in macromolecular systems of infinite size and the shift accounts for a smallest local relaxation rate in macromolecules of finite size. The resulting autocorrelation functions are analytic for any time t ≥ 0. Using results from a molecular dynamics simulation of lysozyme, we demonstrate that the model fits the position autocorrelation functions of the C(α)-atoms exceptionally well and reveals moreover a strong correlation between the residue's solvent-accessible surface and the fitted model parameters.


Morphine sulfate, transdermal fentanyl citrate and ketorolac tromethamine: effects on postoperative pulmonary function.
The physiological effects of morphine sulfate and fentanyl citrate on postoperative pulmonary function demonstrate varying degrees of respiratory depression related to dose, route of administration and pre-existing pathologies. Current postoperative analgesic therapies that include concomitant use of narcotic agonists and nonsteroidal anti-inflammatory drugs require adjustment of drug dosages to provide adequate pain relief while avoiding drug-induced complications. Specific nursing considerations include understanding therapeutic and adverse effects of these agents on pain and ventilation, continuous pain assessment and early recognition and treatment of respiratory depression.


Platelet-derived growth factor signaling through ephrin-b2 regulates hepatic vascular structure and function.
Cirrhosis is associated with prominent changes in sinusoidal structure and function. Although the resident pericyte in liver, the hepatic stellate cell (HSC), is well characterized in the process of fibrogenesis, signaling pathways that regulate HSC vascular function are less developed. Because pericyte populations outside the liver are increasingly being recognized as a key cell type for angiogenesis and changes in vascular structure, in this study, we explore new HSC-signaling pathways that regulate sinusoidal structure and function. Real-time video microscopy and quantitative software analysis of vascular tube formation were used to measure HSC angiogenesis in vitro. Platelet-derived growth factor (PDGF) and ephrin-signaling pathways were modulated using molecular and pharmacologic techniques. Complementary whole animal studies were performed to correlate in vitro findings with pericyte functions in vivo. We show that PDGF promotes a phenotype of HSC evidenced by enhanced HSC-driven vascular tube formation in vitro and enhanced HSC coverage of sinusoids in vivo. This angiogenic phenotype modulates specific pericyte vascular functions including permeability and pressure regulation. Furthermore, we identify a key role for ephrin-B2 as a downstream effector of PDGF signaling. These studies elucidate novel HSC-signaling pathways that regulate microvascular structure and function in liver.


An integrated strategy for transmission control of Schistosoma japonicum in a marshland area of China: findings from a five-year longitudinal survey and mathematical modeling.
In 2004 an aggressive plan was instituted aiming to achieve nationwide transmission control of schistosomiasis by 2015. Here, we report a longitudinal study on the control of schistosomiasis in Anhui province, China. Using a mathematical model, we compared the effects of different control strategies implemented in the study area. During the 5-year study period, a 60.8% reduction in human prevalence was observed from 2005 (7.95%) to 2009 (3.1%), and snail infection decreased from 0.063% in 2005 to zero in 2009. Results of the model agree well with the first 3-year field observations and suggest continuous decrease in human infections in the last 2 years, whereas the last 2-year field observations indicated that human infections appeared to be stable even with continuous control. Our findings showed that the integrated control strategy was effective, and we speculated that other factors besides bovines might contribute to the local transmission of the disease.


Expression and functional characterization of recombinant tributyltin-binding protein type 2.
Tributyltin-binding proteins (TBT-bps) are members of the fish lipocalins that were isolated from the blood of Japanese flounder (Paralichthys olivaceus) and function in the binding and detoxification of TBT. In this study, we constructed a baculovirus-silkworm expression system and obtained recombinant TBT-bp2 (rTBT-bp2; 31.5 kDa) from the hemolymph of silkworm larvae injected with a recombinant baculovirus containing the TBT-bp2 gene. The binding potential of rTBT-bp2 was investigated and compared to that of the previously available recombinant TBT-bp1 (rTBT-bp1). Both rTBT-bp2 and rTBT-bp1 bound to DAUDA, a typical fluorescent ligand of lipocalins, with dissociation constants of 0.97 and 1.75 µM, respectively. The Hill coefficient value indicated that rTBT-bp2 may have multiple binding sites and strong negative cooperativity. These results suggest that the typical central cavity of lipocalins composed of eight specific β-sheets is conserved in rTBT-bp2, as it is in rTBT-bp1, although rTBT-bp2 has different effects than rTBT-bp1 in TBT binding. In a competition assay, rTBT-bp2 displayed exponential binding affinity to TBT with an inhibition constant of 0.29 µM, demonstrating that TBT binds to the central ligand pocket of rTBT-bp2. However, three fatty acids did not show any affinity to rTBT-bp2. Further studies are required to elucidate the endogenous function of TBT-bps as fish lipocalins and their function in responding to xenobiotics.


Lysozyme Mutants Accumulate in Cells while Associated at their N-terminal Alpha-domain with the Endoplasmic Reticulum Chaperone GRP78/BiP.
Amyloidogenic human lysozyme variants deposit in cells and cause systemic amyloidosis. We recently observed that such lysozymes accumulate in the endoplasmic reticulum (ER) with the ER chaperone GRP78/BiP, accompanying the ER stress response. Here we investigated the region of lysozyme that is critical to its association with GRP78/BiP. In addition to the above-mentioned variants of lysozyme, we constructed lysozyme truncation or substitution mutants. These were co-expressed with GRP78/BiP (tagged with FLAG) in cultured human embryonic kidney cells, which were analyzed by western blotting and immunocytochemistry using anti-lysozyme and anti-FLAG antibodies. The amyloidogenic variants were confirmed to be strongly associated with GRP78/BiP as revealed by the co-immunoprecipitation assay, whereas N-terminal mutants pruned of 1-41 or 1-51 residues were found not to be associated with the chaperone. Single amino acid substitutions for the leucine array along the α-helices in the N-terminal region resulted in wild-type lysozyme remaining attached to GRP78/BiP. These mutations also tended to show lowered secretion ability. We conclude that the N-terminal α-helices region of the lysozyme is pivotal for its strong adhesion to GRP78/BiP. We suspect that wild-type lysozyme interacts with the GRP at this region as a step in the proper folding monitored by the ER chaperone.


The contribution of non-physician clinicians to the provision of surgery in rural Zambia-a randomised controlled trial.
The global shortage of surgeons disproportionately impacts low- and middle-income countries. To mitigate this, Zambia introduced a 'task-shifting' solution and started to train non-physician clinicians (NPCs) called medical licentiates (ML) to perform surgery. The aim of this randomised controlled trial was to assess their contribution to the delivery of surgical care in rural hospitals in Zambia. Sixteen hospitals were randomly assigned to intervention and control arms of the study. Nine MLs were deployed to eight intervention sites. Crude numbers of selected major surgical procedures between intervention and control sites were compared before and after the intervention. Volume and outcomes of surgery were compared within intervention hospitals, between NPCs and surgically active medical doctors (MDs). There was a significant increase in the numbers of caesarean sections (CS) in the intervention hospitals (+ 15.2%) and a drop by almost half in the control group (- 47%) (P = 0.015), between the two time periods. There were marginal shifts in the numbers of index procedures: a small drop in the intervention group (- 4.9%) and slight increase in the control arm (+ 4.8%) (P = 0.505). In all pairs, MLs had higher mean number of CS and other major surgical cases done in the intervention period compared with MDs. There was no significant difference in postoperative wound infection rates for CS (P = 0.884) and other major surgical cases (P = 0.33) at intervention hospitals between MLs and MDs. This study provided evidence that the ML training programme in Zambia is an effective and safe way to bridge the gap in rural hospitals between the demand and the limited availability of surgically trained workforce in the country. Such evidence is greatly needed as more developing countries are developing national surgical plans. ISRCTN66099597 Registered: 07/01/2014.


Smoking and inequalities in mortality in 11 European countries: a birth cohort analysis.
To study the trends of smoking-attributable mortality among the low and high educated in consecutive birth cohorts in 11 European countries. Register-based mortality data were collected among adults aged 30 to 79 years in 11 European countries between 1971 and 2012. Smoking-attributable deaths were estimated indirectly from lung cancer mortality rates using the Preston-Glei-Wilmoth method. Rate ratios and rate differences among the low and high-educated were estimated and used to estimate the contribution of inequality in smoking-attributable mortality to inequality in total mortality. In most countries, smoking-attributable mortality decreased in consecutive birth cohorts born between 1906 and 1961 among low- and high-educated men and high-educated women, but not among low-educated women among whom it increased. Relative educational inequalities in smoking-attributable mortality increased among both men and women with no signs of turning points. Absolute inequalities were stable among men but slightly increased among women. The contribution of inequality in smoking-attributable mortality to inequality in total mortality decreased in consecutive generations among men but increased among women. Smoking might become less important as a driver of inequalities in total mortality among men in the future. However, among women, smoking threatens to further widen inequalities in total mortality.


The interactions of nitric oxide and adenosine on penicillin-induced epileptiform activity in rats.
In this study, the influence of nitric oxide (NO) and adenosine systems on penicillin-induced epileptiform activity was examined in rats. NO donor, sodium nitroprusside (SNP, 50 micrograms per rat, i.c.v.) reduced the frequency but not the amplitude of epileptiform discharges. Non-selective NOS inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME, 100 micrograms per rat, i.c.v.) practically did not exert any effect on the spike frequency and amplitude. Adenosine (100 micrograms per rat, i.c.) reduced spike frequency but not the amplitude, whereas theophylline (100 micrograms per rat, i.c.v.) increased the mean spike frequency and amplitude of penicillin-induced epileptiform discharges. Co-injection of theophylline and L-NAME did not cause a further increase in the epileptiform activity compared with theophylline. When NO production was blocked with L-NAME, the inhibitory effects of adenosine were lost. The obtained results suggest that NO and adenosine may decrease penicillin-induced epileptiform activity in rats and that NO, at least in part, may mediate the anticonvulsant effect of adenosine.


Human milk metagenome: a functional capacity analysis.
Human milk contains a diverse population of bacteria that likely influences colonization of the infant gastrointestinal tract. Recent studies, however, have been limited to characterization of this microbial community by 16S rRNA analysis. In the present study, a metagenomic approach using Illumina sequencing of a pooled milk sample (ten donors) was employed to determine the genera of bacteria and the types of bacterial open reading frames in human milk that may influence bacterial establishment and stability in this primal food matrix. The human milk metagenome was also compared to that of breast-fed and formula-fed infants' feces (n = 5, each) and mothers' feces (n = 3) at the phylum level and at a functional level using open reading frame abundance. Additionally, immune-modulatory bacterial-DNA motifs were also searched for within human milk. The bacterial community in human milk contained over 360 prokaryotic genera, with sequences aligning predominantly to the phyla of Proteobacteria (65%) and Firmicutes (34%), and the genera of Pseudomonas (61.1%), Staphylococcus (33.4%) and Streptococcus (0.5%). From assembled human milk-derived contigs, 30,128 open reading frames were annotated and assigned to functional categories. When compared to the metagenome of infants' and mothers' feces, the human milk metagenome was less diverse at the phylum level, and contained more open reading frames associated with nitrogen metabolism, membrane transport and stress response (P < 0.05). The human milk metagenome also contained a similar occurrence of immune-modulatory DNA motifs to that of infants' and mothers' fecal metagenomes. Our results further expand the complexity of the human milk metagenome and enforce the benefits of human milk ingestion on the microbial colonization of the infant gut and immunity. Discovery of immune-modulatory motifs in the metagenome of human milk indicates more exhaustive analyses of the functionality of the human milk metagenome are warranted.


Professionalism: orientation exercises for incoming osteopathic medical students and developing class vision statements.
The Philadelphia College of Osteopathic Medicine has developed an exercise to introduce professional ethics and behavior at the earliest stages of medical education. During orientation, each incoming class creates a class vision statement. After small group discussions on professional ethics, honesty, and responsibilities, representatives from each group collated student input and constructed a class vision statement reflective of student consensus on these issues. Each vision statement was recited as an oath during the white coat ceremony at the conclusion of the orientation program. Despite the fact that previous vision statements were unavailable to each incoming class, there were many commonalities among the statements created. Central elements of all vision statements include commitment to altruism, compassionate treatment of patients, and honesty and integrity in all professional interactions. Humility, the capacity to recognize and accept one's limitations in knowledge and skills, was also a key element in each statement. Three of four statements specifically recognized the teamwork and mutual respect that should be engendered among all members of the health care team. Each vision statement had prominent statements regarding the learning process during osteopathic medical school and acknowledged the importance of active and lifelong learning in the students' career paths. Student evaluation of this exercise has been positive, especially the recitation of the statement during the white coat ceremony. Results suggest that the development of a class vision statement represents a powerful mechanism for addressing the importance of professional attitudes, behaviors, and ethics at the earliest stages of medical education.


Cancer cells exploit eIF4E2-directed synthesis of hypoxia response proteins to drive tumor progression.
Human tumors display considerable diversity in their genetic makeup but share common physiologic attributes such as a hypoxic microenvironment that contribute to the malignant phenotype. Hypoxic cells switch from eukaryotic initiation factor 4E (eIF4E) to eIF4E2 cap-dependent translation to synthesize a portion of their proteins. Here, we show that genetically distinct human cancer cells exploit eIF4E2-directed protein synthesis to form cellular masses larger than approximately 0.15 mm, the diffusion limit of oxygen. Cancer cells depleted of eIF4E2 are indistinguishable from control cells under normoxic conditions, but are unable to survive and proliferate in low oxygen conditions. Activation of eIF4E2-directed translation is essential for cancer cells to form a hypoxic tumor core in in vitro spheroids and to form detectable tumors in in vivo xenograft assays. In contrast, the eIF4E-directed protein synthesis pathway alone cannot sustain cellular adaptation to hypoxia in vitro or confer tumorigenic potential in xenograft assays. These data demonstrate that the phenotypic expression of the cancer genome requires translation by the eIF4E2-directed hypoxic protein synthesis machinery.


A scientific comment on the welfare of domesticated ruminants slaughtered without stunning.
This review provides a scientific comment on the welfare of ruminants slaughtered by ventral-neck incision without stunning. Evidence is derived from studies of calves, sheep and goats. Reference is also made to findings in other mammals including humans. Pain is an inherently subjective experience and only indirect indices are available in animals. Neurophysiological tools are widely used in humans to assess pain and have demonstrated that electroencephalographic (EEG) variables correlate well with subjective evaluations of pain. These neurophysiological tools have also been applied in animal studies. In humans pain is associated with major cutting injuries and it is widely accepted that farm animals also experience pain due to such injuries. Overwhelming international scientific opinion has long been that slaughter by neck incision of conscious animals causes pain. A series of studies in calves demonstrated that slaughter by ventral-neck incision is likely to be perceived as painful. It is proposed that, as in cattle, non-stunned sheep and goats would experience pain in a similar manner. The precise assessment of the point after slaughter at which non-stunned animals become insensible remains a major methodological challenge. In sheep it is at least 2-8 seconds, but may be 8-20 seconds in duration. In cattle the mean duration is similar, but can commonly be extended to longer than 60 seconds with occasional instances of even greater durations. Taken together, these findings indicate that because the slaughter of cattle, sheep and goats by ventral-neck incision without prior stunning is likely to cause pain, this poses a risk to animal welfare.


133Xenon ventilation scan as a functional assessment in bronchiectasis.
Mucus impaction in the airways impairs ventilation and exercise tolerance in patients with bronchiectasis. Parameters for evaluating the ventilatory dynamic change have been limited by variable cofactors. We developed a tool to evaluate the changes directly on images of a ventilation scan. We used a 133Xenon ventilation washout scan to assess the time of half clearance (T1/2) of the regions of interest (ROIs) corrected by that of a control area (CA) as T1/2ROI/CA. We then compared the ventilation washout scan with high-resolution computer tomography (HRCT) scoring to assess the severity of bronchiectasis, as well as conducting 6-minute walking tests or spirometry for the evaluation of the clinical response to a 3-day course of chest physiotherapy. Nine patients with bronchiectasis and mucus hypersecretion were enrolled in this study. The functional impairments by mucus impaction or air trapping were well documented in the ventilation washout scan, which not only provided an anatomical image but also dynamic profiles. The ratios of T1/2ROI/CA were significantly correlated to the corresponding scoring of HRCT (3.45 +/- 0.85 vs 7.50 +/- 1.51, r2 = 0.61, p = 0.023, n = 8). The improvement in T1/2ROI/CA (from 3.45 +/- 0.85 to 2.60 +/- 0.59, p = 0.022, n = 9) was paralleled by an increase in the 6-minute walking test (from 310.4 +/- 43.2 m to 352.4 +/- 45.1 m, p = 0.028, n = 7). The 133Xenon scan may be used to evaluate the heterogeneity of ventilation abnormalities and the efficacy of clinical therapy directly in patients with bronchiectasis.


Prediagnostic Physical Activity and Colorectal Cancer Survival: Overall and Stratified by Tumor Characteristics.
Physical activity is associated with a lower incidence of colorectal cancer; however, the relationship of physical activity with colorectal cancer survival is not yet clear. We evaluated the association between prediagnostic physical activity and colorectal cancer survival, overall and accounting for tumor markers associated with colorectal cancer survival: BRAF and KRAS mutation status and microsatellite instability (MSI) status. Participants were 20- to 74-year-old colorectal cancer patients diagnosed between 1998 and 2007 from the population-based Seattle Colon Cancer Family Registry (S-CCFR). Self-reported physical activity in the years preceding colorectal cancer diagnosis was summarized as average metabolic equivalent task hours per week (MET-h/wk; n = 1,309). Somatic BRAF and KRAS mutations and MSI status were evaluated on a subset of patients (n = 1043). Cox regression was used to estimate HRs and 95% confidence intervals (CI) for overall and disease-specific survival after adjusting for relevant confounders. Stratified analyses were conducted across categories of BRAF, KRAS, and MSI, as well as tumor stage and site. Higher prediagnostic recreational physical activity was associated with significantly more favorable overall survival (HR for highest vs. lowest category, 0.70; 95% CI, 0.52-0.96); associations were similar for colorectal cancer-specific survival. Results consistently indicated a favorable association with physical activity across strata defined by tumor characteristics. Individuals who were physically active before colorectal cancer diagnosis experienced better survival than those who were inactive or minimally active. Our results support existing physical activity recommendations for colorectal cancer patients and suggest that the beneficial effect of activity is not specific to a particular molecular phenotype of colorectal cancer.


Connecting the legs with a spring improves human running economy.
Human running is inefficient. For every 10 calories burned, less than 1 is needed to maintain a constant forward velocity - the remaining energy is, in a sense, wasted. The majority of this wasted energy is expended to support the bodyweight and redirect the center of mass during the stance phase of gait. An order of magnitude less energy is expended to brake and accelerate the swinging leg. Accordingly, most devices designed to increase running efficiency have targeted the costlier stance phase of gait. An alternative approach is seen in nature: spring-like tissues in some animals and humans are believed to assist leg swing. While it has been assumed that such a spring simply offloads the muscles that swing the legs, thus saving energy, this mechanism has not been experimentally investigated. Here, we show that a spring, or 'exotendon', connecting the legs of a human reduces the energy required for running by 6.4±2.8%, and does so through a complex mechanism that produces savings beyond those associated with leg swing. The exotendon applies assistive forces to the swinging legs, increasing the energy optimal stride frequency. Runners then adopt this frequency, taking faster and shorter strides, and reduce the joint mechanical work to redirect their center of mass. Our study shows how a simple spring improves running economy through a complex interaction between the changing dynamics of the body and the adaptive strategies of the runner, highlighting the importance of considering each when designing systems that couple human and machine.


GSTM1 and GSTT1 polymorphisms, cigarette smoking, and risk of colon cancer: a population-based case-control study in North Carolina (United States).
Cigarette smoke is a risk factor for colon cancer, but the importance of dose and interaction with genetic susceptibility remain poorly understood. We used data from a population-based case control study, to examine the association between cigarette smoking and colon cancer in African Americans and whites, and colon cancer and polymorphisms in GSTM1 and GSTT1. A total of 554 cases of primary colon cancer and 874 controls were included in this analysis. We found no association between cigarette smoking (ever versus never) and colon cancer in African Americans (odds ratio (OR)=0.93, 95% confidence interval (CI)=0.65-1.33). In contrast, there was an increased risk of cigarette smoking in whites (OR=1.43, CI=1.05-1.94). There was a small increased risk of colon cancer for individuals with GSTM1 null (African Americans, OR=1.43, CI, 0.98-2.09; whites, OR=1.19, CI, 0.90-1.58) and a decreased risk of colon cancer for individuals with GSTT1 null (African Americans, OR=0.59, CI: 0.40-0.86; whites, OR=0.72, CI: 0.53-1.00). There were weak interactions between GSTT1 null and cigarette smoking in whites, and GSTM1 null genotype and cigarette smoking in African Americans. GSTT1 and GSTM1 polymorphisms may be weakly related to colon cancer risk and there may be racial differences in gene-smoking interactions.


Is intermittent short-course anti-TB regimen as efficient and safe as daily anti-TB regimen for treating childhood TB?
Childhood tuberculosis (TB) is a common clinical condition in low- and middle-income nations like India where TB is endemic. Different guidelines vary in the recommendations on treatment regimes for childhood TB. Apart from clinical outcomes the decision to use one regimen over another also has an implication in the form of health system burden. The evidence summary presents the comparison between the intermittent regimens with the daily anti-TB regimen for childhood TB.


Long-term, low-dose lithium treatment does not impair renal function in the elderly: a 2-year randomized, placebo-controlled trial followed by single-blind extension.
Recent studies evaluated the disease-modifying properties of lithium in mild cognitive impairment and dementia. Although potentially effective for these purposes, chronic lithium use in regard to safety in the elderly needs to be better explored. To evaluate the effect of long-term lithium treatment at subtherapeutic doses on renal function in older adults. Secondary aims were to evaluate the clinical safety and tolerability of this treatment and its effects on thyroid, immune, and glycemic functions. Between February 2007 and October 2011, a 2-year randomized, double-blind, placebo-controlled trial followed by a single-blinded phase for an additional 2 years. Sixty-one patients with mild cognitive impairment (Mayo Clinic criteria) were randomized to receive lithium or placebo. Renal function was estimated by the abbreviated Modification of Diet in Renal Disease (aMDRD) and the Chronic Kidney Disease-Epidemiology study (CKD-EPI) equations. Leukocytes, serum thyroid-stimulating hormone (TSH) and free thyroxine (T₄), and serum glucose and insulin were determined. Tolerability was evaluated at 3-month intervals through systematic clinical examinations and by the UKU Side Effect Rating Scale. Analysis of longitudinal regression indicated that no significant changes in renal function were detected by the aMDRD (P = .453) and CKD-EPI (P = .213) equations after 4 years of lithium treatment. Significant increases in the number of neutrophils (P = .038), serum TSH (P = .034), and body weight (P = .015) were observed in the lithium group. The lithium group presented more overall adverse events (P = .045), particularly interfering in daily activities (P < .001). In addition, those patients had a higher incidence of diabetes mellitus (P = .037) and arrhythmia (P = .028). Chronic use of lithium at low doses did not affect renal function and was clinically safe. However, some other potentially relevant adverse events were observed and others could not be ruled out due to limitations of the study design. ClinicalTrials.gov identifier: NCT01055392.


A recombinant factor VIII A2 domain polypeptide quantitatively neutralizes human inhibitor antibodies that bind to A2.
Human antibodies that inactivate coagulation factor VIII (fVIII), known as inhibitors, have been shown by immunoblotting or immunoprecipitation assays to bind predominantly to epitopes within the A2 and/or C2 domains of the fVIII protein. Because these assays simply measure antibody binding, a soluble recombinant polypeptide containing the fVIII A2 domain was used to develop a quantitative inhibitor neutralization assay for antibodies that bound only to A2 by immunoblotting assay. Antibodies from six of eight inhibitor plasmas were fully neutralized by A2 (> or = 90%), whereas two were only partially neutralized. These results established the fVIII inhibitor properties of anti-A2 antibodies. In immunoprecipitation assays, five of the eight inhibitors also had significant levels of anti-light-chain antibody. In one case, this light-chain antibody was shown to have inhibitor activity. Because it did not bind to the C2 domain, this antibody appears to define a new inhibitor epitope within the fVIII light chain. Another inhibitor, which was partially neutralized by A2, was not neutralized by the light chain, even though it contained anti-light-chain antibodies by immunoprecipitation assay. Our results show additional complexities of the immune response to fVIII.


Bystander activation and anti-tumor effects of CD8+ T cells following Interleukin-2 based immunotherapy is independent of CD4+ T cell help.
We have previously demonstrated that immunotherapy combining agonistic anti-CD40 and IL-2 (IT) results in synergistic anti-tumor effects. IT induces expansion of highly cytolytic, antigen-independent "bystander-activated" (CD8(+)CD44high) T cells displaying a CD25(-)NKG2D(+) phenotype in a cytokine dependent manner, which were responsible for the anti-tumor effects. While much attention has focused on CD4(+) T cell help for antigen-specific CD8(+) T cell expansion, little is known regarding the role of CD4(+) T cells in antigen-nonspecific bystander-memory CD8(+) T cell expansion. Utilizing CD4 deficient mouse models, we observed a significant expansion of bystander-memory T cells following IT which was similar to the non-CD4 depleted mice. Expanded bystander-memory CD8(+) T cells upregulated PD-1 in the absence of CD4(+) T cells which has been published as a hallmark of exhaustion and dysfunction in helpless CD8(+) T cells. Interestingly, compared to CD8(+) T cells from CD4 replete hosts, these bystander expanded cells displayed comparable (or enhanced) cytokine production, lytic ability, and in vivo anti-tumor effects suggesting no functional impairment or exhaustion and were enriched in an effector phenotype. There was no acceleration of the post-IT contraction phase of the bystander memory CD8(+) response in CD4-depleted mice. The response was independent of IL-21 signaling. These results suggest that, in contrast to antigen-specific CD8(+) T cell expansion, CD4(+) T cell help is not necessary for expansion and activation of antigen-nonspecific bystander-memory CD8(+) T cells following IT, but may play a role in regulating conversion of these cells from a central memory to effector phenotype. Additionally, the expression of PD-1 in this model appears to be a marker of effector function and not exhaustion.


An interdisciplinary and catchment approach to enhancing urban flood resilience: a Melbourne case.
This paper presents a novel interdisciplinary and catchment-based approach for exploring urban flood resilience. Our research identified and developed a diverse set of adaptation measures for Elwood, a suburb in Melbourne, Australia, that is vulnerable to pluvial and coastal flooding. We drew on methods from social science, urban design and environmental engineering to gain integrated insights into the opportunities for Elwood to increase its flood resilience and urban liveability. Results showed that an appropriate balance of social, infrastructural and urban design responses would be required to retreat from, accommodate and protect against flood risk. These would also deliver broader benefits such as securing water supplies through harvested stormwater and mitigating extreme heat through greener landscapes. Our interdisciplinary approach demonstrated the value of (i) engaging with the community to understand their concerns, aspirations and adaptation ideas, (ii) exploring design measures that densify and use urban forms in ways that implement adaptation measures while responding to local context, (iii) adopting modelling techniques to test the performance, robustness and economic viability of possible adaptation solutions, and (iv) innovating governance arrangements and principles needed to improve flood resilience in the Elster Creek catchment. Our research also provided valuable insight on how to operationalize interdisciplinary work in practice, highlighting the importance of sharing an impact agenda, taking a place-based approach, developing a common conceptual framework, and fostering a constructive team culture. This article is part of the theme issue 'Urban flood resilience'.


Reassessment of critical lead effects by the German Human Biomonitoring Commission results in suspension of the human biomonitoring values (HBM I and HBM II) for lead in blood of children and adults.
In Germany, the Human Biomonitoring Commission of the Federal Environment Agency continuously assesses environmental pollutants to derive human biomonitoring (HBM) and reference values. HBM values are derived on the basis of toxicological, epidemiological studies or toxicokinetic extrapolation which provides a concentration of a substance or its metabolites corresponding to tolerable intake doses. Two levels are defined: HBM I and HBM II. In 1996, the Commission set a HBM I of 100 microg/l for lead in blood of children < or =12 years and females of a reproductive age and a HBM I of 150 microg/l for the other persons. In the light of findings from epidemiological studies on effects below 100 microg/l, the Commission reevaluated and confirmed the assessment from 1996 in 2002. Meanwhile the general decline in lead pollution has allowed recent studies to include more cohorts with blood lead levels predominantly below 100 microg/l. These data confirm that lead's critical effect, particularly on the developing organism and during early childhood, concerns the nervous system and that negative correlations between blood lead levels and relevant variables (cognitive function, behaviour) occur at blood lead levels below 100 microg/l. The new data also support the possible persistence of lead-induced effects into adulthood. It is not possible to indicate thresholds. Concerning the estimation of the size of the effects, recent studies suggest that lead's influence on development is comparable with other factors influencing development. Furthermore inorganic lead and compounds were classified by IARC in group 2A (probably carcinogenic to humans) and by the German Research Foundation (MAK Commission) in category 2 (to be regarded as human carcinogen). We conclude that any setting of an "effect threshold" for blood lead levels would be arbitrary and therefore unjustified. As a consequence the Commission suspends the HBM values for lead in blood of children and adults.


Performance and physiological responses during a sprint interval training session: relationships with muscle oxygenation and pulmonary oxygen uptake kinetics.
The purpose of this study was to examine the cardiorespiratory and muscle oxygenation responses to a sprint interval training (SIT) session, and to assess their relationships with maximal pulmonary O(2) uptake [Formula: see text], on- and off- [Formula: see text] kinetics and muscle reoxygenation rate (Reoxy rate). Ten male cyclists performed two 6-min moderate-intensity exercises (≈90-95% of lactate threshold power output, Mod), followed 10 min later by a SIT session consisting of 6 × 30-s all out cycling sprints interspersed with 2 min of passive recovery. [Formula: see text] kinetics at Mod onset ([Formula: see text]) and cessation ([Formula: see text]) were calculated. Cardiorespiratory variables, blood lactate ([La](b)) and muscle oxygenation level of the vastus lateralis (tissue oxygenation index, TOI) were recorded during SIT. Percentage of the decline in power output (%Dec), time spent above 90% of [Formula: see text] (t > 90% [Formula: see text]) and Reoxy rate after each sprint were also recorded. Despite a low mean [Formula: see text] (48.0 ± 4.1% of [Formula: see text]), SIT performance was associated with high peak [Formula: see text] (90.4 ± 2.8% of [Formula: see text]), muscle deoxygenation (sprint ΔTOI = -27%) and [La](b) (15.3 ± 0.7 mmol l(-1)) levels. Muscle deoxygenation and Reoxy rate increased throughout sprint repetitions (P < 0.001 for both). Except for t > 90% [Formula: see text] versus [Formula: see text] [r = 0.68 (90% CL, 0.20; 0.90); P = 0.03], there were no significant correlations between any index of aerobic function and either SIT performance or physiological responses [e.g., %Dec vs. [Formula: see text]: r = -0.41 (-0.78; 0.18); P = 0.24]. Present results show that SIT elicits a greater muscle O(2) extraction with successive sprint repetitions, despite the decrease in external power production (%Dec = 21%). Further, our findings obtained in a small and homogenous group indicate that performance and physiological responses to SIT are only slightly influenced by aerobic fitness level in this population.


Investigation of LKB1 Ser431 phosphorylation and Cys433 farnesylation using mouse knockin analysis reveals an unexpected role of prenylation in regulating AMPK activity.
The LKB1 tumour suppressor protein kinase functions to activate two isoforms of AMPK (AMP-activated protein kinase) and 12 members of the AMPK-related family of protein kinases. The highly conserved C-terminal residues of LKB1 are phosphorylated (Ser431) by PKA (cAMP-dependent protein kinase) and RSK (ribosomal S6 kinase) and farnesylated (Cys433) within a CAAX motif. To better define the role that these post-translational modifications play, we created homozygous LKB1S431A/S431A and LKB1C433S/C433S knockin mice. These animals were viable, fertile and displayed no overt phenotypes. Employing a farnesylation-specific monoclonal antibody that we generated, we established by immunoprecipitation that the vast majority, if not all, of the endogenous LKB1 is prenylated. Levels of LKB1 localized at the membrane of the liver of LKB1C433S/C433S mice and their fibroblasts were reduced substantially compared with the wild-type mice, confirming that farnesylation plays a role in mediating membrane association. Although AMPK was activated normally in the LKB1S431A/S431A animals, we unexpectedly observed in all of the examined tissues and cells taken from LKB1C433S/C433S mice that the basal, as well as that induced by the AMP-mimetic AICAR (5-amino-4-imidazolecarboxamide riboside), AMPK activation, phenformin and muscle contraction were significantly blunted. This resulted in a reduced ability of AICAR to inhibit lipid synthesis in primary hepatocytes isolated from LKB1C433S/C433S mice. The activity of several of the AMPK-related kinases analysed [BRSK1 (BR serine/threonine kinase 1), BRSK2, NUAK1 (NUAK family, SNF1-like kinase 1), SIK3 (salt-inducible kinase 3) and MARK4 (MAP/microtubule affinity-regulating kinase 4)] was not affected in tissues derived from LKB1S431A/S431A or LKB1C433S/C433S mice. Our observations reveal for the first time that farnesylation of LKB1 is required for the activation of AMPK. Previous reports have indicated that a pool of AMPK is localized at the plasma membrane as a result of myristoylation of its regulatory AMPKβ subunit. This raises the possibility that LKB1 farnesylation and myristoylation of AMPKβ might promote the interaction and co-localization of these enzymes on a two-dimensional membrane surface and thereby promote efficient activation of AMPK.


Selection Processing in Noun and Verb Production in Left- and Right-Sided Parkinson's Disease Patients.
Verbs are more difficult to produce than nouns. Thus, if executive resources are reduced as in Parkinson's disease (PD), verbs are penalized compared to nouns. However, in an experimental condition in which it is the noun that must be selected from a larger number of alternatives compared to the verb, it is the noun production that becomes slower and more prone to errors. Indeed, patients are slower and less accurate than normal subjects when required to produce nouns from verbs (VN) in a morphology derivation tasks (e.g., "osservazione" from "osservare") ["observation" from "observe"] than verbs from nouns in a morphology generation task, in which only a verb can be generated from the noun (NV) (e.g., "fallire" from "fallimento") ["to fail" from "failure"]. In the Italian language morphology, in fact, generation and derivation tasks differ in the number of lexical entries among which the response must be selected. The left Inferior Frontal Gyrus (IFG) has been demonstrated to be involved in selection processes. In the present study, we explored if the ability to select words is related to the cortical thickness of the left IFG. Twelve right-sided PD with nigrostriatal hypofunctionality in the left hemisphere (RPD-LH), 9 left-sided PD with nigrostriatal hypofunctionality in the right hemisphere (LPD-RH) and 19 healthy controls (HC) took part in the study. NV and VN production tasks were administered; accuracy and reaction times (RTs) were collected. All 40 subjects received a structural MRI examination. Cortical thickness of the IFG and volumetric measurements for subcortical regions, thought to support selection processes, were computed using FreeSurfer. In VN derivation tasks RPD-LH patients were less accurate than LPD-RH patients (accuracy: 66% vs. 77%). No difference emerged among the three groups in RTs. Task accuracy/RTs and IFG thickness showed a significant correlation only in RPD-LH. Not only nouns (as expected) but also verbs were correlated with cortical thickness. This suggests that the linguistic nature of the stimuli along with executive resources are both relevant during word selection processes. Our data confirm that executive resources and language interact in the left IFG in word production tasks.


Relationship between circulating n-3 fatty acid concentrations and endothelial function in early adulthood.
Fish consumption is inversely associated with cardiovascular mortality, presumably because of n-3 fatty acids in fish. Whether the protection of n-3 fatty acids extends beyond clinical coronary disease to influence the early vascular biology of atherosclerosis remains unclear. This study determined whether circulating levels of n-3 fatty acids are associated with vascular endothelial function in early adulthood. Three hundred and twenty-six adults (157 males, 169 females, aged 20 to 28 years) had high-resolution ultrasound measurements of flow-mediated brachial artery dilatation (FMD) (endothelium-dependent) and arterial response to glyceryl trinitrate (endothelium-independent). Levels of the n-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid in plasma and erythrocyte membranes of subjects were measured. n-3 Fatty acid levels were not related to vascular function in the whole group. In smokers, however, n-3 fatty acids were positively related to flow-mediated dilatation (plasma DHA vs. FMD: 0.045 mm. %(-1), 95% CI 0.011 to 0.079, P=0.01). Flow-mediated dilatation was also associated with n-3 fatty acid levels in subjects in the top third of the insulin, glucose and triglyceride distributions. In young smokers and those with higher fasting insulin, glucose or triglyceride concentrations (factors associated with endothelial dysfunction), n-3 fatty acid levels were positively associated with flow-mediated dilatation. This raises the possibility that physiological levels of circulating n-3 fatty acids may protect the endothelium from early adulthood.


Solid-state fermentation: a promising microbial technology for secondary metabolite production.
Solid state (substrate) fermentation (SSF) has been used successfully for the production of enzymes and secondary metabolites. These products are associated with the stationary phase of microbial growth and are produced on an industrial scale for use in agriculture and the treatment of disease. Many of these secondary metabolites are still produced by submerged liquid fermentations (SmF) even though production by this method has been shown to be less efficient than SSF. As large-scale production increases further, so do the costs and energy demands. SSF has been shown to produce a more stable product, requiring less energy, in smaller fermenters, with easier downstream processing measures. In this article we review an important area of biotechnology, since the recent evidence indicates that bacteria and fungi, growing under SSF conditions, are more than capable of supplying the growing global demand for secondary metabolites.


Simultaneous measurement of blood flow and glucose metabolism by autoradiographic techniques.
A double tracer autoradiographic technique using 131I-iodo-antipyrine and 14C-deoxyglucose is presented for the simultaneous measurement of blood flow and cerebral glucose utilization in the same animal. 131I is a gamma emitting isotope with a half life of 8.06 days and can be detected with adequate resolution on standard autoradiographic films. Autoradiograms are made before and after decay of 131I; the time interval between the 2 exposures and the concentration of the 2 tracers is adjusted to avoid significant cross-contamination. In this way, 2 film exposures are obtained which can be processed quantitatively like single tracer autoradiograms. The validity of the method for the investigation of local coupling of flow and metabolism was tested under various physiological and pathophysiological conditions. Coupling was tight in barbiturate-anesthetized healthy animals, but not under halothane anesthesia where uncoupling occurred in various subcortical structures. Focal seizures induced by topical application of penicillin on the cortical surface led to a coupled increase of metabolism and flow in thalamic relay nuclei but not at the site of penicillin administration where increased glucose utilization was not accompanied by similar increase in blood flow. Both coupled and uncoupled increases in local glucose utilization were observed in spreading depression and in circumscribed areas of experimental brain tumors. The results obtained demonstrate that double tracer autoradiography allows allows the very precise local assessment of cerebral blood flow and glucose utilization, and, therefore, is particularly suited to the study of regional coupling processes under various experimental conditions.


CCN1 triggers adaptive autophagy in cardiomyocytes to curb its apoptotic activities.
Autophagy occurs at basal levels for cellular homeostasis under normal conditions and is increased in response to nutrient starvation or stress to ensure cell survival. However, excessive autophagy can be deleterious to cardiomyocytes. CCN1/Cyr61, a matricellular protein, is expressed in the stressed heart to induce cardiomyopathy. The role of autophagy in CCN1-associated cardiotoxicity was not clear. Here, we found that autophagy was induced in the myocardium of the isoproterenol (ISO; 100 mg/kg/day for 5 days; s.c.) treated mice, where CCN1 expression is colocalized. The knock-in mice carrying an integrin α6β1-binding-defective mutant allele Ccn1-dm were resistant to the ISO-induced cardiac injury and autophagy. Our in vitro studies demonstrated that CCN1 dose- and time-dependently induced GFP-LC3-labeled autophagosome formation in rat cardiomyoblast H9c2 cells. The formation of autolysosomes in response to CCN1 (5 μg/ml; 3 h) treatment was identified by the acridine orange staining. The autophagy induction was confirmed by the elevated protein levels of Beclin 1, Atg5, and LC3-II, and the decrease of p62. Inhibition of autophagy by 3-methyladenine or by silencing Atg5 gene enabled CCN1-induced apoptosis in H9c2 cells, suggesting a protective role of autophagy. CCN1 binds to integrin α6β1 to induce autophagy through reactive oxygen species, and the activation of ERK and JNK. Furthermore, mitophagy was observed after CCN1 treatment for the clearance of depolarized mitochondria. Together, these results demonstrated that autophagy is induced in response to CCN1/α6β1 signaling in cardiomyocytes to alleviate CCN1-associated cardiotoxicity.