Studies on the inhibitory mechanism of iodonium compounds with special reference to neutrophil NADPH oxidase.
Diphenyleneiodonium (DPI) and its analogues have been previously shown to react via a radical mechanism whereby an electron is abstracted from a nucleophile to form a radical, which then adds back to the nucleophile to form covalent adducts [Banks (1966) Chem. Rev. 66, 243-266]. We propose that the inhibition of neutrophil NADPH oxidase by DPI occurs via a similar mechanism. A reduced redox centre in the oxidase could serve as electron donor to DPI, and inhibition would occur after direct phenylation of the redox cofactor, or of adjacent amino acid groups by the DPI radical. In the absence of an activatory stimulus, human neutrophil NADPH-oxidase was not inhibited by DPI. The Ki for time-dependent inhibition by DPI of human neutrophil membrane NADPH oxidase was found to be 5.6 microM. Inhibitory potency of DPI was shown to be directly related to rate of enzyme turnover, indicating the need for a reduced redox centre. Adducts were formed between photoreduced flavin (FAD or FMN) and inhibitor (DPI or diphenyliodonium). These were separated by h.p.l.c. and characterized by absorbance spectroscopy, 1H-n.m.r. and fast-atom-bombardment m.s. and found to have properties consistent with substituted 4a,5-dihydroflavins. After incubation of pig neutrophil membranes with DPI, the quantity of recoverable intact flavin was greatly diminished when NADPH was present to initiate oxidase turnover, indicating that the flavin may be the site of DPI activation. These results may provide a common mechanism of action for iodonium compounds as inhibitors of other flavoenzymes.


Renal function of gene-targeted mice lacking both SGK1 and SGK3.
Serum- and glucocorticoid-inducible kinase (SGK) 1 and SGK3 share the ability to upregulate several ion channels, including the epithelial Na(+) channel. Whereas SGK1 is under genomic control of mineralocorticoids and glucocorticoids, SGK3 is constitutively expressed. The SKG1-knockout (sgk1(-/-)) mouse is seemingly normal when it is fed a standard diet, but its ability to retain NaCl is impaired when it is fed a salt-deficient diet. In the SGK3-knockout (sgk3(-/-)) mouse fed standard and salt-deficient diets, hair growth is strikingly delayed but NaCl excretion is normal. Thus the possibility was considered that SGK1 and SGK3 could mutually replace each other, thus preventing severe NaCl loss in sgk1(-/-) and sgk3(-/-) mice. We crossed SGK1- and SGK3-knockout mice and compared renal electrolyte excretion of the double mutants (sgk1(-/-)/sgk3(-/-)) with that of their wild-type littermates (sgk1(+/+)/sgk3(+/+)). Similar to sgk3(-/-) mice, the sgk1(-/-)/sgk3(-/-) mice display delayed hair growth. Blood pressure was slightly, but significantly (P < 0.03), lower in sgk1(-/-)/sgk3(-/-) (102 +/- 4 mmHg) than in sgk1(+/+)/sgk3(+/+) (114 +/- 3 mmHg) mice, a difference that was maintained in mice fed low- and high-salt diets. Plasma aldosterone concentrations were significantly (P < 0.01) higher in sgk1(-/-)/sgk3(-/-) than in sgk1(+/+)sgk3(+/+) mice fed control (511 +/- 143 vs. 143 +/- 32 pg/ml) and low-salt (1,325 +/- 199 vs. 362 +/- 145 pg/ml) diets. During salt depletion, absolute and fractional excretions of Na(+) were significantly (P < 0.01) higher in sgk1(-/-)/sgk3(-/-) (1.2 +/- 0.2 micromol/24 h g body wt, 0.12 +/- 0.03%) than in sgk1(+/+)/sgk3(+/+) (0.4 +/- 0.1 micromol/24 h g body wt, 0.04 +/- 0.01%) mice. The sgk1(-/-)/sgk3(-/-) mice share the delayed hair growth with sgk3(-/-) mice and the modestly impaired renal salt retention with sgk1(-/-) mice. Additional lack of the isoform kinase does not substantially compound the phenotype for either property.


Lactobacilli for prevention of urogenital infections: a review.
Urogenital infections are a worldwide shared problem that represent the most common reason for a woman to decide to visit to gynaecologist or urologist. The origin of the uropathogens in uncomplicated urinary tract infection and bacterial vaginosis is the fecal flora. Key element of pathogenesis namely the ability of the pathogens to survive exposure to the microflora that exists on the external urogenitalia, in which lactobacilli predominate. Some health food appear to contain > or = 1 common Lactobacillus strain; L. rhamnosus GR-1 was found to be the best of a group of 34 Lactobacillus strains isolated from dairy, poultry, health food. Recently has been reported the first clinical evidence that probiotic lactobacilli can be delivered to the vagina following oral intake. These L. strains possess the ability to adhere to and colonize tissues and the capacity to inhibit the pathogenesis of disease-causing organisms that make them effective probiotic agents. In particularly, two strains, Lactobacillus GG and Lactobacillus rhamnosus GR-1 appear to be effective at colonizing and protecting the intestine and urogenital tract, respectively, against microbial infection. Treating and preventing urogenital infection by instillating probiotic organisms has great appeal to patients and caregivers. The ability to administer orally L. rhamnosus GR-1 and L. fermentum RC-14, which colonize the intestine and vagina, provides a major step in the right direction for patients as it potentially allows for the self administration of therapy.


Spatial analysis of agri-environmental policy uptake and expenditure in Scotland.
Agri-environment is one of the most widely supported rural development policy measures in Scotland in terms of number of participants and expenditure. It comprises 69 management options and sub-options that are delivered primarily through the competitive 'Rural Priorities scheme'. Understanding the spatial determinants of uptake and expenditure would assist policy-makers in guiding future policy targeting efforts for the rural environment. This study is unique in examining the spatial dependency and determinants of Scotland's agri-environmental measures and categorised options uptake and payments at the parish level. Spatial econometrics is applied to test the influence of 40 explanatory variables on farming characteristics, land capability, designated sites, accessibility and population. Results identified spatial dependency for each of the dependent variables, which supported the use of spatially-explicit models. The goodness of fit of the spatial models was better than for the aspatial regression models. There was also notable improvement in the models for participation compared with the models for expenditure. Furthermore a range of expected explanatory variables were found to be significant and varied according to the dependent variable used. The majority of models for both payment and uptake showed a significant positive relationship with SSSI (Sites of Special Scientific Interest), which are designated sites prioritised in Scottish policy. These results indicate that environmental targeting efforts by the government for AEP uptake in designated sites can be effective. However habitats outside of SSSI, termed here the 'wider countryside' may not be sufficiently competitive to receive funding in the current policy system.


Predicting Bevirimat resistance of HIV-1 from genotype.
Maturation inhibitors are a new class of antiretroviral drugs. Bevirimat (BVM) was the first substance in this class of inhibitors entering clinical trials. While the inhibitory function of BVM is well established, the molecular mechanisms of action and resistance are not well understood. It is known that mutations in the regions CS p24/p2 and p2 can cause phenotypic resistance to BVM. We have investigated a set of p24/p2 sequences of HIV-1 of known phenotypic resistance to BVM to test whether BVM resistance can be predicted from sequence, and to identify possible molecular mechanisms of BVM resistance in HIV-1. We used artificial neural networks and random forests with different descriptors for the prediction of BVM resistance. Random forests with hydrophobicity as descriptor performed best and classified the sequences with an area under the Receiver Operating Characteristics (ROC) curve of 0.93 +/- 0.001. For the collected data we find that p2 sequence positions 369 to 376 have the highest impact on resistance, with positions 370 and 372 being particularly important. These findings are in partial agreement with other recent studies. Apart from the complex machine learning models we derived a number of simple rules that predict BVM resistance from sequence with surprising accuracy. According to computational predictions based on the data set used, cleavage sites are usually not shifted by resistance mutations. However, we found that resistance mutations could shorten and weaken the alpha-helix in p2, which hints at a possible resistance mechanism. We found that BVM resistance of HIV-1 can be predicted well from the sequence of the p2 peptide, which may prove useful for personalized therapy if maturation inhibitors reach clinical practice. Results of secondary structure analysis are compatible with a possible route to BVM resistance in which mutations weaken a six-helix bundle discovered in recent experiments, and thus ease Gag cleavage by the retroviral protease.


Multilaboratory Comparison of Pneumococcal Multiplex Immunoassays Used in Immunosurveillance of Streptococcus pneumoniae across Europe.
Surveillance studies are required to estimate the impact of pneumococcal vaccination in both children and the elderly across Europe. The World Health Organization (WHO) recommends use of enzyme immunoassays (EIAs) as standard methods for immune surveillance of pneumococcal antibodies. However, as levels of antibodies to multiple serotypes are monitored in thousands of samples, a need for a less laborious and more flexible method has evolved. Fluorescent-bead-based multiplex immunoassays (MIAs) are suitable for this purpose. An increasing number of public health and diagnostic laboratories use MIAs, although the method is not standardized and no international quality assessment scheme exists. The EU Pneumo Multiplex Assay Consortium was initiated in 2013 to advance harmonization of MIAs and to create an international quality assessment scheme. In a multilaboratory comparison organized by the consortium, agreement among nine laboratories that used their own optimized MIA was assessed on a panel of 15 reference sera for 13 pneumococcal serotypes with the new WHO standard 007sp. Agreement was assessed in terms of assay accuracy, reproducibility, repeatability, precision, and bias. The results indicate that the evaluated MIAs are robust and reproducible for measurement of vaccine-induced antibody responses. However, some serotype-specific variability in the results was observed in comparisons of polysaccharides from different sources and of different conjugation methods, especially for serotype 4. On the basis of the results, the consortium has contributed to the harmonization of MIA protocols to improve reliability of immune surveillance of Streptococcus pneumoniaeIMPORTANCE Serology of Streptococcus pneumoniae is challenging due to existence of multiple clinically relevant serotypes and the introduction of multivalent vaccines in national immunization programs. Multiplex immunoassays (MIAs) are applied as high-throughput cost-effective methods for serosurveillance, and yet laboratories use their own protocols. The aims of this study were to assess the agreement of results generated by MIAs in different laboratories within the EU Pneumo Multiplex Assay Consortium, to analyze factors contributing to differences in outcome, and to create a harmonized protocol. The study demonstrated good agreement of results of MIAs performed by laboratories using controlled assays for determination of levels of vaccine-induced pneumococcal antibodies. The EU Pneumo Multiplex Assay Consortium is open to everyone working in public health services, and it aims to facilitate efforts by participants to run and maintain a cost-effective, reproducible, high-quality MIA platform.


Heat shock transcription factor 1 localizes to sex chromatin during meiotic repression.
Heat shock factor 1 (HSF1) is an important transcription factor in cellular stress responses, cancer, aging, and developmental processes including gametogenesis. Disruption of Hsf1, together with another HSF family member, Hsf2, causes male sterility and complete lack of mature sperm in mice, but the specific role of HSF1 in spermatogenesis has remained unclear. Here, we show that HSF1 is transiently expressed in meiotic spermatocytes and haploid round spermatids in mouse testis. The Hsf1(-/-) male mice displayed regions of seminiferous tubules containing only spermatogonia and increased morphological abnormalities in sperm heads. In search for HSF1 target genes, we identified 742 putative promoters in mouse testis. Among them, the sex chromosomal multicopy genes that are expressed in postmeiotic cells were occupied by HSF1. Given that the sex chromatin mostly is repressed during and after meiosis, it is remarkable that HSF1 directly regulates the transcription of sex-linked multicopy genes during postmeiotic repression. In addition, our results show that HSF1 localizes to the sex body prior to the meiotic divisions and to the sex chromocenter after completed meiosis. To the best of our knowledge, HSF1 is the first known transcription factor found at the repressed sex chromatin during meiosis.


Recombinant growth hormone therapy in patients with ischemic cardiomyopathy : effects on hemodynamics, left ventricular function, and cardiopulmonary exercise capacity.
We studied the effects of recombinant growth hormone (rhGH) on exercise capacity and cardiac function in patients with ischemic cardiomyopathy. Seven patients (aged 55+/-9 years) with mild to moderate congestive heart failure (ejection fraction 31+/-4%) who were on standard therapy were included. The patients were studied at baseline, after 3 months of rhGH treatment, and 3 months after rhGH discontinuation. Cardiac function was assessed by exercise capacity, right heart catheterization at rest and after submaximal exercise, MRI, echocardiography, and Holter monitoring. When administered at a dose of 2 IU/d, rhGH doubled the serum concentration of insulin-like growth factor-I. rhGH improved clinical symptoms and exercise capacity significantly (New York Heart Association class 2.4+/-0.5 initially versus 1.4+/-0.5 at 3 months [mean+/-SD], P<0.05; VO2max 13.6+/-3.8 versus 17.4+/-5.4 mL. kg-1. min-1, P<0.05). Additionally, pulmonary capillary wedge pressures at rest and after submaximal exercise were reduced significantly. Cardiac output increased, particularly at rest (5.0+/-1.1 versus 5.8+/-1.3 L/min; P<0.05). Posterior wall thickness was increased (1.08+/-0.1 versus 1. 24+/-0.3 cm; P<0.05), and the end-diastolic and end-systolic volume indexes decreased significantly after rhGH treatment. There was no significant increase in left ventricular ejection fraction. The improvements were partially reversed 3 months after rhGH discontinuation. The administration of rhGH for 3 months in patients with ischemic cardiomyopathy results in significant improvement in hemodynamics and clinical function. The attenuation of left ventricular remodeling persisted 3 months after discontinuation of treatment.


Pervasive shifts in forest dynamics in a changing world.
Forest dynamics arise from the interplay of environmental drivers and disturbances with the demographic processes of recruitment, growth, and mortality, subsequently driving biomass and species composition. However, forest disturbances and subsequent recovery are shifting with global changes in climate and land use, altering these dynamics. Changes in environmental drivers, land use, and disturbance regimes are forcing forests toward younger, shorter stands. Rising carbon dioxide, acclimation, adaptation, and migration can influence these impacts. Recent developments in Earth system models support increasingly realistic simulations of vegetation dynamics. In parallel, emerging remote sensing datasets promise qualitatively new and more abundant data on the underlying processes and consequences for vegetation structure. When combined, these advances hold promise for improving the scientific understanding of changes in vegetation demographics and disturbances.


Salvia officinalis (Sage) Leaf Extract as Add-on to Statin Therapy in Hypercholesterolemic Type 2 Diabetic Patients: a Randomized Clinical Trial.
The efficacy and safety of Salvia officinalis combined with statin have not been evaluated in dyslipidemic diabetes mellitus type 2 (DDMT2) so far. The plant extract antioxidant activity was determined by the DPPH radical scavenging assay. The total flavonoid, total phenolic and quercetin contents of the capsules containing the plant extract were also measured. Moreover, the effects of 2-month extract intake (500 mg capsule three times a day) as add-on to daily use of 15 mg glyburide, 2000 mg metformin and 10 mg atorvastatin on the blood levels of fasting glucose (FG), 2 h postprandial glucose (2hPPG), glycosylated hemoglobin (HbA1c), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglyceride, high-density lipoprotein cholesterol (HDL-C), serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), creatinine and body mass index were studied in 50 patients and compared with the placebo group (n=50).The extract IC50 in the DPPH assay was 87.26±0.003 µg/mL (mean±SD), whereas the ascorbic acid IC50 was 5.626± 0.001 µg/mL (mean±SD). The total flavonoid, total phenolic and quercetin contents of the capsule containing the plant extract were 39.76±3.58 mg of rutin equivalents (mean±SD), 30.33±1.23 mg of gallic acid (mean±SD) and 0.13 mg, respectively. The extract lowered FG, 2hPPG, HbA1c, TC, LDL-C and triglyceride levels, but increased HDL-C level compared to the placebo at the endpoint (P<0.05). The extract did not affect the other parameters significantly and no adverse effect was reported. The extract has substantial antioxidant activity which may be beneficial for the prevention of the cardiovascular complications of DDMT2. Moreover, addition of the extract to statin therapy is apparently safe and further improves lipid profile.


Faster cognitive decline in elders without dementia and decreased risk of cancer mortality: NEDICES Study.
To assess whether faster cognitive decline in elders without dementia is associated with decreased risk of cancer mortality. In this population-based, prospective study of 2,627 people without dementia aged 65 years and older (Neurological Disorders in Central Spain), a 37-item version of the Mini-Mental State Examination (37-MMSE) was administered at 2 visits (baseline and follow-up, approximately 3 years later). We divided change in 37-MMSE into tertiles (lower tertile ≥ 2 point improvement in score, higher tertile ≥ 2 point decline in score). Community-dwelling elders were followed for a median of 12.9 years, after which the death certificates of those who died were examined. A total of 1,003 (38.2%) died, including 339 (33.8%) deaths among participants who were in the higher tertile of 37-MMSE change and 664 (66.2%) deaths among those in the remaining tertiles. Cancer was reported significantly less often in those in the higher tertile of MMSE change (20.6%) than in those in the remaining tertiles (28.6%): in an unadjusted Cox model, hazard ratio for cancer mortality in participants within the higher tertile = 0.75 (p = 0.04) compared with the participants within the remaining tertiles. In a Cox model that adjusted for a variety of demographic factors and comorbidities, hazard ratio for cancer mortality in participants within the higher tertile = 0.70 (p = 0.01). In this population-based, prospective study of community-dwelling elders without dementia, faster cognitive decline was associated with a decreased risk of cancer mortality. Further studies are required to elucidate this inverse association in elders without dementia.


Application of in vitro biotransformation data and pharmacokinetic modeling to risk assessment.
The adverse biological effects of toxic substances are dependent upon the exposure concentration and the duration of exposure. Pharmacokinetic models can quantitatively relate the external concentration of a toxicant in the environment to the internal dose of the toxicant in the target tissues of an exposed organism. The exposure concentration of a toxic substance is usually not the same as the concentration of the active form of the toxicant that reaches the target tissues following absorption, distribution, and biotransformation of the parent toxicant. Biotransformation modulates the biological activity of chemicals through bioactivation and detoxication pathways. Many toxicants require biotransformation to exert their adverse biological effects. Considerable species differences in biotransformation and other pharmacokinetic processes can make extrapolation of toxicity data from laboratory animals to humans problematic. Additionally, interindividual differences in biotransformation among human populations with diverse genetics and lifestyles can lead to considerable variability in the bioactivation of toxic chemicals. Compartmental pharmacokinetic models of animals and humans are needed to understand the quantitative relationships between chemical exposure and target tissue dose as well as animal to human differences and interindividual differences in human populations. The data-based compartmental pharmacokinetic models widely used in clinical pharmacology have little utility for human health risk assessment because they cannot extrapolate across dose route or species. Physiologically based pharmacokinetic (PBPK) models allow such extrapolations because they are based on anatomy, physiology, and biochemistry. In PBPK models, the compartments represent organs or groups of organs and the flows between compartments are actual blood flows. The concentration of a toxicant in a target tissue is a function of the solubility of the toxicant in blood and tissues (partition coefficients), blood flow into the tissue, metabolism of the toxicant in the tissue, and blood flow out of the tissue. The appropriate degree of biochemical detail can be added to the PBPK models as needed. Comparison of model simulations with experimental data provides a means of hypothesis testing and model refinement. In vitro biotransformation data from studies with isolated liver cells or subcellular fractions from animals or humans can be extrapolated to the intact organism based upon protein content or cell number. In vitro biotransformation studies with human liver preparations can provide quantitative data on human interindividual differences in chemical bioactivation. These in vitro data must be integrated into physiological models to understand the true impact of interindividual differences in chemical biotransformation on the target organ bioactivation of chemical contaminants in air and drinking water.


Inhibition of PIK3 signaling pathway members by the ovotoxicant 4-vinylcyclohexene diepoxide in rats.
4-Vinylcyclohexene diepoxide (VCD), an occupational chemical that specifically destroys primordial and small primary follicles in the ovaries of rats and mice, is thought to target an oocyte-expressed tyrosine kinase receptor, Kit. This study compared the temporal effect of VCD on protein distribution of KIT and its downstream PIK3-activated proteins, AKT and FOXO3. Postnatal Day 4 Fischer 344 rat ovaries were cultured in control media ± VCD (30 μM) for 2-8 days (d2-d8). KIT, AKT, phosphorylated AKT, FOXO3, and pFOXO3 protein levels were assessed by Western blotting and/or immunofluorescence staining with confocal microscopy. Phosphorylated AKT was decreased (P < 0.05) in oocyte nuclei in primordial (39% decrease) and small primary (37% decrease) follicles within 2 days of VCD exposure. After d4, VCD reduced (P < 0.05) oocyte staining for KIT (primordial, 44% decrease; small primary, 39% decrease) and FOXO3 (primordial, 40% decrease; small primary, 36% decrease) protein. Total AKT and pFOXO3 were not affected by VCD at any time. Akt1 mRNA, as measured by quantitative RT-PCR, was reduced (P < 0.05) by 23% on d4 of VCD exposure, but returned to control levels on d6 and d8. VCD exposure reduced Foxo3a mRNA by 26% on d6 (P < 0.05) and by 23% on d8 (P < 0.1). These results demonstrate that the earliest observed effect of VCD is an inhibition of phosphorylation and nuclear localization of AKT in the oocyte of primordial and small primary follicles. This event is followed by reductions in KIT and FOXO3 protein subcellular distribution prior to changes in mRNA. Thus, these findings further support that VCD induces ovotoxicity by directly targeting the oocyte through posttranslational inhibition of KIT-mediated signaling components.


Very extensive nonmaternal care predicts mother-infant attachment disorganization: Convergent evidence from two samples.
We examined whether a maximum threshold of time spent in nonmaternal care exists, beyond which infants have an increased risk of forming a disorganized infant-mother attachment. The hours per week infants spent in nonmaternal care at 7-8 months were examined as a continuous measure and as a dichotomous threshold (over 40, 50 and 60 hr/week) to predict infant disorganization at 12-15 months. Two different samples (Austin and NICHD) were used to replicate findings and control for critical covariates: mothers' unresolved status and frightening behavior (assessed in the Austin sample, N = 125), quality of nonmaternal caregiving (assessed in the NICHD sample, N = 1,135), and family income and infant temperament (assessed in both samples). Only very extensive hours of nonmaternal care (over 60 hr/week) and mothers' frightening behavior independently predicted attachment disorganization. A polynomial logistic regression performed on the larger NICHD sample indicated that the risk of disorganized attachment exponentially increased after exceeding 60 hr/week. In addition, very extensive hours of nonmaternal care only predicted attachment disorganization after age 6 months (not prior). Findings suggest that during a sensitive period of attachment formation, infants who spend more than 60 hr/week in nonmaternal care may be at an increased risk of forming a disorganized attachment.


Estimation of the warfarin dose with clinical and pharmacogenetic data.
Genetic variability among patients plays an important role in determining the dose of warfarin that should be used when oral anticoagulation is initiated, but practical methods of using genetic information have not been evaluated in a diverse and large population. We developed and used an algorithm for estimating the appropriate warfarin dose that is based on both clinical and genetic data from a broad population base. Clinical and genetic data from 4043 patients were used to create a dose algorithm that was based on clinical variables only and an algorithm in which genetic information was added to the clinical variables. In a validation cohort of 1009 subjects, we evaluated the potential clinical value of each algorithm by calculating the percentage of patients whose predicted dose of warfarin was within 20% of the actual stable therapeutic dose; we also evaluated other clinically relevant indicators. In the validation cohort, the pharmacogenetic algorithm accurately identified larger proportions of patients who required 21 mg of warfarin or less per week and of those who required 49 mg or more per week to achieve the target international normalized ratio than did the clinical algorithm (49.4% vs. 33.3%, P<0.001, among patients requiring < or = 21 mg per week; and 24.8% vs. 7.2%, P<0.001, among those requiring > or = 49 mg per week). The use of a pharmacogenetic algorithm for estimating the appropriate initial dose of warfarin produces recommendations that are significantly closer to the required stable therapeutic dose than those derived from a clinical algorithm or a fixed-dose approach. The greatest benefits were observed in the 46.2% of the population that required 21 mg or less of warfarin per week or 49 mg or more per week for therapeutic anticoagulation.


Screening for mild cognitive impairment: a systematic review.
Patients with mild cognitive impairment account for a significant number of referrals to old age psychiatry services and specialist memory clinics. The cognitive evaluation of such patients is commonly restricted to brief dementia screens, with no consideration to their suitability for assessing MCI. Here, we review the utility of such cognitive screens for MCI and provide an overview of validated instruments. We identified papers published after Petersen and colleagues 1999 MCI criteria (Petersen et al., 1999) and examining face-to-face cognitive screening for MCI from publication databases using combinations of the search terms 'mild cognitive impairment' and 'cognitive screening'. We also combined the former search with the names of 39 screening tests recently identified in a relevant review (Cullen et al., 2007). Fifteen cognitive screening instruments were identified, 11 cover a restricted range of cognitive domains. High sensitivity and specificity for MCI relative to healthy controls were reported for two comprehensive and two noncomprehensive screening instruments, adequate test-retest and inter-rater reliability for only one of these. With the exception of three studies, sample sizes were universally small (i.e. n </= 100), and prognostic values were reported for only two of the identified 15 screening measures. Sensitivities of the full domain measures were universally high, but information about their specificity against psychiatric and non-progressive neurological conditions and predictive validity is lacking. Several cognitive screening instruments afford the clinician the ability to detect MCI, early AD, and in some cases non-AD dementia, but they cannot currently be used to make reliable inferences about the course and eventual outcome of MCI.


Interleukin-6 response of epithelial cell lines to bacterial stimulation in vitro.
This study demonstrated that epithelial cell lines secrete interleukin-6 (IL-6) in response to stimulation with gram-negative bacteria. Human epithelial cell lines of urinary tract origin (A-498 and J82) and of intestinal origin (HT-29 and Caco-2) were analyzed for the secretion of IL-6 by using the B9 bioassay. The supernatants from cells maintained with culture medium were used to assess the constitutive production of IL-6. The supernatants from cells exposed to Escherichia coli strains, lipopolysaccharide, lipid A, and isolated fimbriae were used to quantitate the IL-6 response to these stimulants. The urinary tract epithelial cell lines were found to constitutively secrete IL-6. The IL-6 activity in the supernatants of the bladder cell line (J82) increased above constitutive levels after 2 h of stimulation by most of the bacterial strains tested. The IL-6 activity in the supernatants of the kidney line (A-498) accumulated at a constant rate over the 24-h assay period. The role of bacterial adherence for the induction of IL-6 production was investigated by comparing the responses to recombinant E. coli strains expressing different fimbriae. In addition, isolated P and S fimbriae with and without the receptor binding domain were also used as stimulants. The IL-6 activity in the supernatants of the bladder cell line increased after exposure to bacteria and bacterial products regardless of their adhesive properties. In contrast, the kidney cell line was stimulated to secrete significantly more IL-6 by adhering bacteria and by adhesin-positive P fimbriae than by nonadhering bacteria or adhesin-negative P fimbriae. The S-fimbrial preparations had no specific effects on the IL-6 activity of the cell supernatants. These results are consistent with our hypothesis that epithelial cells can be a major source of IL-6 when stimulated by bacteria and that the adhesive properties of the bacteria can influence this response.


Mode of action of membrane-disruptive lytic compounds from the marine dinoflagellate Alexandrium tamarense.
Certain allelochemicals of the marine dinoflagellate Alexandrium tamarense cause lysis of a broad spectrum of target protist cells but the lytic mechanism is poorly defined. We first hypothesized that membrane sterols serve as molecular targets of these lytic compounds, and that differences in sterol composition among donor and target cells may cause insensitivity of Alexandrium and sensitivity of targets to lytic compounds. We investigated Ca(2+) influx after application of lytic fractions to a model cell line PC12 derived from a pheochromocytoma of the rat adrenal medulla to establish how the lytic compounds affect ion flux associated with lysis of target membranes. The lytic compounds increased permeability of the cell membrane for Ca(2+) ions even during blockade of Ca(2+) channels with cadmium. Results of a liposome assay suggested that the lytic compounds did not lyse such target membranes non-specifically by means of detergent-like activity. Analysis of sterol composition of isolates of A. tamarense and of five target protistan species showed that both lytic and non-lytic A. tamarense strains contain cholesterol and dinosterol as major sterols, whereas none of the other tested species contain dinosterol. Adding sterols and phosphatidylcholine to a lysis bioassay with the cryptophyte Rhodomonas salina for evaluation of competitive binding indicated that the lytic compounds possessed apparent high affinity for free sterols and phosphatidylcholine. Lysis of protistan target cells was dose-dependently reduced by adding various sterols or phosphatidylcholine. For three tested sterols, the lytic compounds showed highest affinity towards cholesterol followed by ergosterol and brassicasterol. Cholesterol comprised a higher percentage of total sterols in plasma membrane fractions of A. tamarense than in corresponding whole cell fractions. We conclude therefore that although the molecular targets of the lytic compounds are likely to involve sterol components of membranes, A. tamarense must have a complex self-protective mechanism that still needs to be addressed.


Special health considerations in African-American elders.
Older African Americans constitute an expanding part of the elderly population in the United States. Although socioeconomic factors affect longevity and functional status more than race, African-American elders, as a whole, show poorer health status, as well as greater levels of financial strain and care-giver burden. Incidence rates of hypertension, heart disease, stroke, end-stage renal disease, dementia and prostate cancer are higher among African Americans than among the white population. The incidence of depression, however, is lower. Cancer survival rates are also lower, in part because of lower rates of cancer screening in this group. Physicians should carefully choose instruments to assess cognitive and physical status in African-American elders. The Activities of Daily Living scale and the Short Portable Mental Status Questionnaire are two tools that have been specially tested and shown to be reliable and valid in this population group. The Geriatric Depression Scale is a useful diagnostic tool that is quick to use in a busy office practice. Taking the time during an initial visit to understand the patient's values and perceptions of health and illness builds a sense of comfort and trust that will set a positive tone for the entire doctor-patient relationship and may empower the patient to take positive steps to improve health habits.


Energy expenditure at rest and during walking in patients with chronic respiratory failure: a prospective two-phase case-control study.
Measurements of Energy Expenditure (EE) at rest (REE) and during physical activities are increasing in interest in chronic patients. In this study we aimed at evaluating the validity/reliability of the SenseWear®Armband (SWA) device in terms of REE and EE during assisted walking in Chronic Respiratory Failure (CRF) patients receiving long-term oxygen therapy (LTOT). In a two-phase prospective protocol we studied 40 severe patients and 35 age-matched healthy controls. In phase-1 we determined the validity and repeatability of REE measured by SWA (REEa) in comparison with standard calorimetry (REEc). In phase-2 we then assessed EE and Metabolic Equivalents-METs by SWA during the 6-minute walking test while breathing oxygen in both assisted (Aid) or unassisted (No-Aid) modalities. When compared with REEc, REEa was slightly lower in patients (1351±169 vs 1413±194 kcal/day respectively, p<0.05), and less repeatable than in healthy controls (0.14 and 0.43 coefficient respectively). COPD patients with CRF patients reported a significant gain with Aid as compared with No-Aid modality in terms of meters walked, perceived symptoms and EE. SWA provides a feasible and valid method to assess the energy expenditure in CRF patients on LTOT, and it shows that aided walking results in a substantial energy saving in this population.


Clathrin terminal domain-ligand interactions regulate sorting of mannose 6-phosphate receptors mediated by AP-1 and GGA adaptors.
Clathrin plays important roles in intracellular membrane traffic including endocytosis of plasma membrane proteins and receptors and protein sorting between the trans-Golgi network (TGN) and endosomes. Whether clathrin serves additional roles in receptor recycling, degradative sorting, or constitutive secretion has remained somewhat controversial. Here we have used acute pharmacological perturbation of clathrin terminal domain (TD) function to dissect the role of clathrin in intracellular membrane traffic. We report that internalization of major histocompatibility complex I (MHCI) is inhibited in cells depleted of clathrin or its major clathrin adaptor complex 2 (AP-2), a phenotype mimicked by application of Pitstop® inhibitors of clathrin TD function. Hence, MHCI endocytosis occurs via a clathrin/AP-2-dependent pathway. Acute perturbation of clathrin also impairs the dynamics of intracellular clathrin/adaptor complex 1 (AP-1)- or GGA (Golgi-localized, γ-ear-containing, Arf-binding protein)-coated structures at the TGN/endosomal interface, resulting in the peripheral dispersion of mannose 6-phosphate receptors. By contrast, secretory traffic of vesicular stomatitis virus G protein, recycling of internalized transferrin from endosomes, or degradation of EGF receptor proceeds unperturbed in cells with impaired clathrin TD function. These data indicate that clathrin is required for the function of AP-1- and GGA-coated carriers at the TGN but may be dispensable for outward traffic en route to the plasma membrane.


Validating a transnational fracture treatment registry using a standardized method.
Subsequent to a three-month pilot phase, recruiting patients for the newly established BFCC (Baltic Fracture Competence Centre) transnational fracture registry, a validation of the data quality needed to be carried out, applying a standardized method. During the literature research, the method of "adaptive monitoring" fulfilled the requirements of the registry and was applied. It consisted of a three-step audit process; firstly, scoring of the overall data quality, followed by source data verification of a sample size, relative to the scoring result, and finally, feedback to the registry on measures to improve data quality. Statistical methods for scoring of data quality and visualisation of discrepancies between registry data and source data were developed and applied. Initially, the data quality of the registry scored as medium. During source data verification, missing items in the registry, causing medium data quality, turned out to be absent in the source as well. A subsequent adaptation of the score evaluated the registry's data quality as good. It was suggested to add variables to some items in order to improve the accuracy of the registry. The application of the method of adaptive monitoring has only been published by Jacke et al., with a similar improvement of the scoring result following the audit process. Displaying data from the registry in graphs helped to find missing items and discover issues with data formats. Graphically comparing the degree of agreement between the registry and source data allowed to discover systematic faults. The method of adaptive monitoring gives a substantiated guideline for systematically evaluating and monitoring a registry's data quality and is currently second to none. The resulting transparency of the registry's data quality could be helpful in annual reports, as published by most major registries. As the method has been rarely applied, further successive applications in established registries would be desirable.


The effects of passive leg press training on jumping performance, speed, and muscle power.
Passive leg press (PLP) training was developed based on the concepts of the stretch-shortening cycle (SSC) and the benefits of high muscle contraction velocity. Passive leg press training enables lower limb muscle groups to apply a maximum downward force against a platform moved up and down at high frequency by an electric motor. Thus, these muscle groups accomplished both concentric and eccentric isokinetic contractions in a passive, rapid, and repetitive manner. This study investigates the effects of 10 weeks of PLP training at high and low movement frequencies have on jumping performance, speed, and muscle power. The authors selected 30 college students who had not performed systematic resistance training in the previous 6 months, including traditional resistance training at a squat frequency of 0.5 Hz, PLP training at a low frequency of 0.5 Hz, and PLP training at a high frequency of 2.5 Hz, and randomly divided them into 3 groups (n = 10). The participants' vertical jump, drop jump, 30-m sprint performance, explosive force, and SSC efficiency were tested under the same experimental procedures at pre- and post-training. Results reveal that high-frequency PLP training significantly increased participants' vertical jump, drop jump, 30-m sprint performance, instantaneous force, peak power, and SSC efficiency (p < 0.05). Additionally, their change rate abilities were substantially superior to those of the traditional resistance training (p < 0.05). The low-frequency PLP training significantly increased participants' vertical jump, 30-m sprint performance, instantaneous force, and peak power (p < 0.05). However, traditional resistance training only increased participants' 30-m sprint performance and peak power (p < 0.05). The findings suggest that jump performance, speed, and muscle power significantly improved after 10 weeks of PLP training at high movement frequency. A PLP training machine powered by an electrical motor enables muscles of the lower extremities to contract faster compared with voluntary contraction. Therefore, muscle training with high contraction velocity is one of the main methods of increasing muscle power. Passive leg press training is a unique method for enhancing jump performance, speed, and muscle power.


Management of right ventricular myxoma diagnosed at full-term pregnancy.
The diagnosis of cardiac myxoma in a woman at term pregnancy is extremely rare. Prompt surgical removal of the tumor is generally advised because of the high risk of potentially fatal complications. On the other hand, cardiac surgery during pregnancy is a delicate procedure which carries a significantly increased maternal risk when performed at or immediately after delivery. A previously healthy 23-year-old woman at 38 weeks' gestation was diagnosed with myxoma in the right ventricle on the basis of clinical and echocardiographic examination. The patient went into labor while awaiting urgent Cesarean section. Severe right heart failure developed which completely resolved after delivery of a healthy baby. After balancing the risks of uncontrolled uterine bleeding associated with cardiopulmonary bypass against those of thromboembolism and valvular obstruction associated with the presence of myxoma itself, emergency cardiac surgery was rejected. The tumor was successfully removed five days after Cesarean section and the patient recovered uneventfully. Cesarean section should be done as soon as possible. Considering the increased maternal morbidity and mortality when delivery is immediately followed by cardiopulmonary bypass, urgent cardiac surgery may be more reasonable than an emergency one for a patient who is clinically stable and at low risk of thromboembolism.


Curcumin enhances the apoptosis-inducing potential of TRAIL in prostate cancer cells: molecular mechanisms of apoptosis, migration and angiogenesis.
We have recently shown that curcumin (a diferuloylmethane) inhibits growth and induces apoptosis, and also demonstrated that TRAIL induces apoptosis by binding to specific cell surface death receptors in prostate cancer cells. The objectives of this paper were to investigate the molecular mechanisms by which curcumin enhanced the apoptosis-inducing potential of TRAIL in prostate cancer cells. Curcumin enhanced the apoptosis-inducing potential of TRAIL in androgen-unresponsive PC-3 cells and sensitized androgen-responsive TRAIL-resistant LNCaP cells. Curcumin inhibited the expressions of Bcl-2, Bcl-XL, survivin and XIAP, and induced the expressions Bax, Bak, PUMA, Bim, and Noxa and death receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5) in both cell lines. Overexpression of dominant negative FADD inhibited the interactive effects of curcumin and TRAIL on apoptosis. Treatment of these cells with curcumin resulted in activation of caspase-3, and caspase-9, and drop in mitochondrial membrane potential, and these events were further enhanced when combined with TRAIL. Curcumin inhibited capillary tube formation and migration of HUVEC cells and these effects were further enhanced in the presence of MEK1/2 inhibitor PD98059. The ability of curcumin to inhibit capillary tube formation and cell migration, and enhance the therapeutic potential of TRAIL suggests that curcumin alone or in combination with TRAIL can be used for prostate cancer prevention and/or therapy.


Tissue oxygen saturation and outcome after cardiac surgery.
Cardiopulmonary bypass during cardiac surgery can result in a shortfall in oxygen delivery relative to demand, marked by a decrease in muscle tissue oxygen saturation as blood flow is redistributed to vital organs. Such "tissue shock" might impair postoperative recovery. To determine the association of changes in tissue oxygen saturation with postoperative outcome in cardiac surgery patients. In 74 adults undergoing cardiac surgery, tissue oxygen saturation in the thenar eminence was recorded using near-infrared spectroscopy before and during induction of anesthesia, throughout surgery, and in the intensive care unit until extubation or for a maximum monitoring time of 24 hours. The measurements were related to postoperative outcome. Mean tissue oxygen saturation increased from 81.7% to 88.5% with induction of anesthesia and decreased to 78.9% and 69.9% during surgery and on arrival in the intensive care unit, respectively. Saturation increased to 77.8% by 6 hours after surgery and remained stable. Mean saturation during the first minutes of anesthesia and 20 minutes in the intensive care unit was lower in patients with a postoperative morbidity than in patients without such morbidity on day 15 (81.1% vs 87.6%; P = .04) and on day 3 (72.9% vs 85.5%; P = .009). No associations with other outcome measures were observed. In patients undergoing cardiac surgery, reduced tissue oxygen saturation in the thenar eminence may be associated with poor postoperative outcome. Further studies are needed to confirm these findings and to determine whether measures to improve the balance between oxygen delivery and consumption might improve both tissue oxygen saturation and outcome.


A census of membrane-bound and intracellular signal transduction proteins in bacteria: bacterial IQ, extroverts and introverts.
Analysis of complete microbial genomes showed that intracellular parasites and other microorganisms that inhabit stable ecological niches encode relatively primitive signaling systems, whereas environmental microorganisms typically have sophisticated systems of environmental sensing and signal transduction. This paper presents results of a comprehensive census of signal transduction proteins--histidine kinases, methyl-accepting chemotaxis receptors, Ser/Thr/Tyr protein kinases, adenylate and diguanylate cyclases and c-di-GMP phosphodiesterases--encoded in 167 bacterial and archaeal genomes, sequenced by the end of 2004. The data have been manually checked to avoid false-negative and false-positive hits that commonly arise during large-scale automated analyses and compared against other available resources. The census data show uneven distribution of most signaling proteins among bacterial and archaeal phyla. The total number of signal transduction proteins grows approximately as a square of genome size. While histidine kinases are found in representatives of all phyla and are distributed according to the power law, other signal transducers are abundant in certain phylogenetic groups but virtually absent in others. The complexity of signaling systems differs even among closely related organisms. Still, it usually can be correlated with the phylogenetic position of the organism, its lifestyle, and typical environmental challenges it encounters. The number of encoded signal transducers (or their fraction in the total protein set) can be used as a measure of the organism's ability to adapt to diverse conditions, the 'bacterial IQ', while the ratio of transmembrane receptors to intracellular sensors can be used to define whether the organism is an 'extrovert', actively sensing the environmental parameters, or an 'introvert', more concerned about its internal homeostasis. Some of the microorganisms with the highest IQ, including the current leader Wolinella succinogenes, are found among the poorly studied beta-, delta- and epsilon-proteobacteria. Among all bacterial phyla, only cyanobacteria appear to be true introverts, probably due to their capacity to conduct oxygenic photosynthesis, using a complex system of intracellular membranes. The census data, available at http://www.ncbi.nlm.nih.gov/Complete_Genomes/SignalCensus.html, can be used to get an insight into metabolic and behavioral propensities of each given organism and improve prediction of the organism's properties based solely on its genome sequence.


Health information needs of visually impaired people: a systematic review of the literature.
Access to, and provision of, information is key to reducing health inequalities in health and social care. However, information is not always accessible and does not always meet the needs of specific groups. One such group is people who are visually impaired. The aim of the present study was to identify the health information needs of visually impaired people, and highlight major gaps in the literature. A systematic review of the literature was undertaken following, where possible, the NHS Centre for Reviews and Dissemination framework. Modifications included the sources searched, the critical appraisal checklist used (Critical Skills Training in Appraisal for Librarians) and the method of data synthesis. Out of the 1114 references identified, only 16 met the inclusion criteria. Quality of reporting of the literature was poor, and this must be taken into consideration when interpreting the findings of this review. The majority of studies were concerned with information for healthy living (e.g. health promotion). The focus of the remaining studies was on information about visual impairment or coping with visual impairment, and about accessing health services (e.g. medication labels and appointment letters). The majority of studies conducted to date have concentrated on the format of information. There are surprisingly few empirical studies which have examined the health information needs of this group. A number of gaps in the literature have been identified. These relate to the types of health information, non-format aspects of information (e.g. content and timing), sources of health and social care information, treating visually impaired people as a heterogeneous group, and recognising the value of actively involving visually impaired people in the research process. Although this review has identified a number of implications for practice, the paucity of evidence places a heavier onus on future research. Visually impaired people are likely to have unique health and social care information needs which are worthy of exploration in their own right.


Response control networks are selectively modulated by attention to rare events and memory load regardless of the need for inhibition.
Recent evidence has sparked debate about the neural bases of response selection and inhibition. In the current study, we employed two reactive inhibition tasks, the Go/Nogo (GnG) and Simon tasks, to examine questions central to these debates. First, we investigated whether a fronto-cortical-striatal system was sensitive to the need for inhibition per se or the presentation of infrequent stimuli, by manipulating the proportion of trials that do not require inhibition (Go/Compatible trials) relative to trials that require inhibition (Nogo/Incompatible trials). A cortico-subcortical network composed of insula, putamen, and thalamus showed greater activation on salient and infrequent events, regardless of the need for inhibition. Thus, consistent with recent findings, key parts of the fronto-cortical-striatal system are engaged by salient events and do not appear to play a selective role in response inhibition. Second, we examined how the fronto-cortical-striatal system is modulated by working memory demands by varying the number of stimulus-response (SR) mappings. Right inferior parietal lobule showed decreasing activation as the number of SR mappings increased, suggesting that a form of associative memory - rather than working memory - might underlie performance in these tasks. A broad motor planning and control network showed similar trends that were also modulated by the number of motor responses required in each task. Finally, bilateral lingual gyri were more robustly engaged in the Simon task, consistent with the role of this area in shifts of visuo-spatial attention. The current study sheds light on how the fronto-cortical-striatal network is selectively engaged in reactive control tasks and how control is modulated by manipulations of attention and memory load.


TNF-alpha in asthma.
Although only 5-10% of patients with asthma are relatively unresponsive to treatment with inhaled corticosteroids, refractory asthma represents an important condition, as these patients suffer considerable morbidity and mortality and consume a disproportionately large amount of health resource. Treatment options are limited and there is a large unmet clinical need for additional therapies. Tumour necrosis factor (TNF)-alpha is a pro-inflammatory cytokine that has been implicated in many aspects of the airway pathology in asthma, and which has recently been highlighted as potentially important in refractory asthma. The development of neutralising biological agents against TNF-alpha has allowed us to test the role of this cytokine in vivo. Preliminary studies have demonstrated an improvement in lung function, airway hyperresponsiveness and asthma quality-of-life, together with a reduction in exacerbation frequency, in patients treated with anti-TNF-alpha therapy.


Therapeutic drug repositioning using personalized proteomics of liquid biopsies.
In patients with limited response to conventional therapeutics, repositioning of already approved drugs can bring new, more effective options. Current drug repositioning methods, however, frequently rely on retrospective computational analyses and genetic testing - time consuming methods that delay application of repositioned drugs. Here, we show how proteomic analysis of liquid biopsies successfully guided treatment of neovascular inflammatory vitreoretinopathy (NIV), an inherited autoinflammatory disease with otherwise poor clinical outcomes. Vitreous biopsies from NIV patients were profiled by an antibody array for expression of 200 cytokine-signaling proteins. Non-NIV controls were compared with NIV samples from various stages of disease progression. Patterns were identified by 1-way ANOVA, hierarchical clustering, and pathway analysis. Subjects treated with repositioned therapies were followed longitudinally. Proteomic profiles revealed molecular pathways in NIV pathologies and implicated superior and inferior targets for therapy. Anti-VEGF injections resolved vitreous hemorrhages without the need for vitrectomy surgery. Methotrexate injections reversed inflammatory cell reactions without the side effects of corticosteroids. Anti-IL-6 therapy prevented recurrent fibrosis and retinal detachment where all prior antiinflammatory interventions had failed. The cytokine array also showed that TNF-α levels were normal and that corticosteroid-sensitive pathways were absent in fibrotic NIV, helping explain prior failure of these conventional therapeutic approaches. Personalized proteomics can uncover highly personalized therapies for autoinflammatory disease that can be timed with specific pathologic activities. This precision medicine strategy can also help prevent delivery of ineffective drugs. Importantly, proteomic profiling of liquid biopsies offers an endpoint analysis that can directly guide treatment using available drugs.


Parental warmth and psychiatric disorders among Puerto Rican children in two different socio-cultural contexts.
Parental warmth (PW) has a strong influence on child development and may precede the onset of psychiatric disorders in children. PW is interconnected with other family processes (e.g., coercive discipline) that may also influence the development of psychiatric disorders in children. We prospectively examined the association between PW and child psychiatric disorders (anxiety, major depression disorder, ADHD, disruptive behavior disorders) over the course of three years among Puerto Rican youth, above and beyond the influence of other family factors. Boricua Youth Study participants, Puerto Rican children 5 to 13 years of age at Wave 1 living in the South Bronx (New York) (SB) and San Juan and Canguas (PR) (n = 2,491), were followed for three consecutive years. Youth psychiatric disorders were measured by the Diagnostic Interview Schedule for Children-IV (DISC-IV). Generalized Linear Mixed models tested the association between PW (Wave 1) and psychiatric disorders in the next two years adjusting for demographic characteristics and family processes. Higher levels of PW were related to lower odds of child anxiety and major depressive disorder over time (OR = 0.69[0.60; 0.79]; 0.49[0.41; 0.58], respectively). The strength of the association between PW and ADHD and disruptive behavior disorder declined over time, although it was still significant in the last assessment (OR = 0.44[0.37; 0.52]; 0.46[0.39; 0.54], respectively). PW had a unique influence on psychiatric disorders beyond the influence of other parenting and family processes. Stronger associations were observed among girls for depression and ADHD. Incorporating PW behaviors such as acceptance, support, and comforting into interventions focused on parenting skills may help prevent child psychiatric disorders.


Markers for context-responsiveness: Client baseline interpersonal problems moderate the efficacy of two psychotherapies for generalized anxiety disorder.
To follow-up a randomized clinical trial that compared the acute and long-term efficacy of 15 sessions of cognitive-behavioral therapy (CBT) versus CBT integrated with motivational interviewing (MI) for severe generalized anxiety disorder (GAD; Westra, Constantino, & Antony, 2016), we (a) characterized the sample's baseline interpersonal problems, and (b) analyzed the role of several theory-relevant problems as moderators of the comparative treatment effects on outcome. We first compared clients' (N = 85) baseline interpersonal problems profile to a general clinical sample. We next conducted piecewise, 2-level growth models to analyze the interactive effects of treatment condition and the hypothesized interpersonal problem indices of nonassertiveness (ranging from low to high), exploitability (ranging from low to high on this specific combination of nonassertiveness and friendliness), and overall agency (ranging from more problems of being too submissive to more problems of being too domineering, including friendly or hostile variants) on acute and follow-up worry reduction. Finally, we conducted hierarchical generalized linear models to examine these interactive effects on the likelihood of achieving clinically meaningful worry reduction across follow-up. As expected, the GAD clients evidenced more nonassertive and exploitable interpersonal problems than the general clinical sample. Also as predicted, clients with more problematic nonassertiveness and low overall agency in their relationships had greater follow-up worry reduction in MI-CBT versus CBT, including to a clinically significant degree for the agency by treatment interaction. GAD-specific interpersonal problems can serve as contextual markers for integrative treatment selection and planning. (PsycINFO Database Record


Hepatotoxic effects of estradiol-17 beta-D-glucuronide in the rat and monkey.
The steroid D-ring glucuronide conjugate estradiol-17 beta-D-glucuronide (E217G) but not the A-ring conjugate E23-glucuronide (E23G) has been shown to inhibit bile flow in the rat. To determine if primates are also sensitive to E217G-induced cholestasis, a noninvasive approach with the use of indocyanine green (ICG), a dye eliminated primarily by biliary excretion, was first validated in the rat and then applied to the rhesus monkey. ICG (16 mg/kg rat; 4 mg/kg monkey) was administered i.v. 10 min after an i.v. bolus dose of either E217G (5.5 or 11 mumol/kg), E23G (11 mumol/kg) or vehicle alone. In the rat, the elimination T1/2 of ICG was increased by the 11 mumol/kg dose of E217G (P less than .025), whereas E23G produced no significant change from vehicle control values. In the monkey, the 5.5 and 11 mumol/kg doses of E217G increased the T1/2 of ICG in a dose-related manner (P less than .005), whereas E23G was without effect. Plasma levels of total radioactivity demonstrated dose-dependent kinetics after the administration of a tracer dose and 11 mumol/kg of [3H]E217G. A rebound of plasma radioactivity was seen at 11 mumol/kg of [3H] E217G, the time course of which mimicked the time course of E217G-induced cholestasis. High-performance liquid chromatographic analysis of rat bile and plasma after the administration of [3H]E217G revealed primarily E217G and estradiol-3-sulfate-17 beta-D-glucuronide together with small amounts of three unidentified metabolites. In the monkey, only E217G and estradiol-3-sulfate-17 beta-D-glucuronide were observed in the plasma after the administration of [3H]E217G. In contrast to E217G, estradiol-3-sulfate-17 beta-D-glucuronide was choleretic in the bile duct-cannulated rat model. These data indicate that E217G is hepatotoxic in both rodents and nonhuman primates.


Convergence of multiple autophagy and cytoplasm to vacuole targeting components to a perivacuolar membrane compartment prior to de novo vesicle formation.
Under starvation conditions, the majority of intracellular degradation occurs at the lysosome or vacuole by the autophagy pathway. The cytoplasmic substrates destined for degradation are packaged inside unique double-membrane transport vesicles called autophagosomes and are targeted to the lysosome/vacuole for subsequent breakdown and recycling. Genetic analyses of yeast autophagy mutants, apg and aut, have begun to identify the molecular machinery as well as indicate a substantial overlap with the biosynthetic cytoplasm to vacuole targeting (Cvt) pathway. Transport vesicle formation is a key regulatory step of both pathways. In this study, we characterize the putative compartment from which both autophagosomes and the analogous Cvt vesicles may originate. Microscopy analyses identified a perivacuolar membrane as the resident compartment for both the Apg1-Cvt9 signaling complex, which mediates the switching between autophagic and Cvt transport, and the autophagy/Cvt-specific phosphatidylinositol 3-kinase complex. Furthermore, the perivacuolar compartment designates the initial site of membrane binding by the Apg/Cvt vesicle component Aut7, the Cvt cargo receptor Cvt19, and the Apg conjugation machinery, which functions in the de novo formation of vesicles. Biochemical isolation of the vesicle component Aut7 and density gradient analyses recapitulate the microscopy findings although also supporting the paradigm that components required for vesicle formation and packaging concentrate at subdomains within the donor membrane compartment.


High Mg effective incorporation in Al-rich AlxGa1 - xN by periodic repetition of ultimate V/III ratio conditions.
According to first-principles calculations, the solubility of Mg as a substitute for Ga or Al in AlxGa1 - xN bulk is limited by large, positive formation enthalpies. In contrast to the bulk case, the formation enthalpies become negative on AlxGa1 - xN surface. In addition, the N-rich growth atmosphere can also be favorable to Mg incorporation on the surface by changing the chemical potentials. On the basis of these special features, we proposed a modified surface engineering technique that applies periodical interruptions under an ultimate V/III ratio condition (extremely N-rich), to enhance Mg effective incorporation. By optimizing the interruption conditions (2 nm interruption interval with 2 s interruption time), the enhancement ratio can be up to about 5 in the Al0.99Ga0.01N epilayer.


Amazon basin: a system in equilibrium.
Despite the very active deforestation of the last decade, the Amazon Basin is still primarily covered with trees and is a system in equilibrium. The Andes form a barrier at the western end of the basin and, coupled with the prevailing easterly winds, ensure an almost unique precipitation and water-recycling regime. On average 50 percent of the precipitation is recycled, and in some areas even more. The soils are poor. Most of the nitrogen and phosphorus is found in the soil, and the remaining nutrient elements are found in the standing biomass. There is some nutrient recycling and little loss from the intact ecosystem, and the small input of nutrients from precipitation maintains a small positive nutrient balance. Continued large-scale deforestation is likely to lead to increased erosion and water runoff with initial flooding in the lower Amazon, together with reduced evapotranspiration and ultimately reduced precipitation. Reduced precipitation in the Amazon could increase the tendency toward continentality and adversely affect climate and the present agriculture in south-central Brazil.


Prophylactic antibiotic therapy for chronic obstructive pulmonary disease (COPD).
There has been renewal of interest in the use of prophylactic antibiotics to reduce the frequency of exacerbations and improve quality of life in chronic obstructive pulmonary disease (COPD). To determine whether or not regular treatment of COPD patients with prophylactic antibiotics reduces exacerbations or affects quality of life. We searched the Cochrane Airways Group Trials Register and bibliographies of relevant studies. The latest literature search was August 2013. Randomised controlled trials (RCTs) that compared prophylactic antibiotics with placebo in patients with COPD. We used the standard methods of The Cochrane Collaboration. Data were extracted and analysed by two independent review authors. Seven RCTs involving 3170 patients were included in this systematic review. All studies were published between 2001 and 2011. Five studies were of continuous antibiotics and two studies were of intermittent antibiotic prophylaxis (termed 'pulsed' for this review). The antibiotics investigated were azithromycin, erythromycin, clarithromycin and moxifloxacin. Azithromycin, erythromycin and clarithromycin are macrolides while moxifloxacin is a fourth-generation synthetic fluoroquinolone antibacterial agent. The study duration varied from three months to 36 months and all used intention-to-treat analysis. Most of the results were of moderate quality. The risk of bias of the included studies was generally low, and we did not downgrade the quality of evidence for risk of bias.The trials recruited participants with a mean age of 66 years and with at least a moderate severity of COPD. Three trials included participants with frequent exacerbations and two trials recruited participants requiring systemic steroids or antibiotics, or both, or who were at the end stage of their disease and required oxygen.The primary outcomes for this review were the number of exacerbations and quality of life.With use of continuous prophylactic antibiotics the number of patients experiencing an exacerbation was reduced (odds ratio (OR) 0.55; 95% confidence interval (CI) 0.39 to 0.77, 3 studies, 1262 participants, high quality). This represented a reduction from 69% of participants in the control group compared to 54% in the treatment group (95% CI 46% to 63%) and the number needed to treat to prevent one exacerbation (NNTb) was therefore 8 (95% CI 5 to 18). The frequency of exacerbations was also reduced with continuous prophylactic antibiotic treatment (rate ratio 0.73; 95% CI 0.58 to 0.91).Use of pulsed antibiotic treatment showed a non-significant reduction in the number of people with exacerbations (OR 0.87; 95% CI 0.69 to 1.09, 1 study, 1149 participants, moderate quality) and the test for interaction showed that this result was significantly different from the effect on exacerbations with continuous antibiotics.There was a statistically significant improvement in quality of life with both continuous and pulsed antibiotic treatment but this was smaller than the four unit improvement that is regarded as being clinically significant (MD -1.78; 95% CI -2.95 to -0.61, 2 studies, 1962 participants, moderate quality).Neither pulsed nor continuous antibiotics showed a significant effect on the secondary outcomes of frequency of hospital admissions, change in lung function, serious adverse events or all-cause mortality (moderate quality evidence).The adverse events that were recorded varied among the trials depending on the different antibiotics used. Azithromycin was associated with a significant hearing loss in the treatment group. The moxifloxacin pulsed study reported a significantly higher number of adverse events in the treatment arm due to the marked increase in gastrointestinal adverse events (P < 0.001). Some adverse events that led to drug discontinuation, such as development of long QTc or tinnitus, were not significantly more frequent in the treatment group than the placebo group but pose important considerations in clinical practice.The development of antibiotic resistance in the community is of major concern. One study found newly colonised patients to have higher rates of antibiotic resistance. Patients colonised with moxifloxacin-sensitive pseudomonas at initiation of therapy rapidly became resistant with the quinolone treatment. Use of continuous prophylactic antibiotics results in a clinically significant benefit in reducing exacerbations in COPD patients. All trials of continuous antibiotics used macrolides hence the noted benefit applies only to the use of continuous macrolide antibiotics. The impact of pulsed antibiotics remains uncertain and requires further research.The trials in this review included patients who were frequent exacerbators and needed treatment with antibiotics or systemic steroids, or who were on supplemental oxygen. There were also older individuals with a mean age of 66 years. The results of these trials apply only to the group of patients who were studied in these trials and may not be generalisable to other groups.Because of concerns about antibiotic resistance and specific adverse effects, consideration of prophylactic antibiotic use should be mindful of the balance between benefits to individual patients and the potential harms to society created by antibiotic overuse.


Cost and cost-effectiveness of newborn home visits: findings from the Newhints cluster-randomised controlled trial in rural Ghana.
Every year, 2·9 million newborn babies die worldwide. A meta-analysis of four cluster-randomised controlled trials estimated that home visits by trained community members in programme settings in Ghana and south Asia reduced neonatal mortality by 12% (95% CI 5-18). We aimed to estimate the costs and cost-effectiveness of newborn home visits in a programme setting. We prospectively collected detailed cost data alongside the Newhints trial, which tested the effect of a home-visits intervention in seven districts in rural Ghana and showed a reduction of 8% (95% CI -12 to 25%) in neonatal mortality. The intervention consisted of a package of home visits to pregnant women and their babies in the first week of life by community-based surveillance volunteers. We calculated incremental cost-effectiveness ratios (ICERs) with Monte Carlo simulation and one-way sensitivity analyses and characterised uncertainty with cost-effectiveness planes and cost-effectiveness acceptability curves. We then modelled the potential cost-effectiveness for baseline neonatal mortality rates of 20-60 deaths per 1000 livebirths with use of a meta-analysis of effectiveness estimates. In the 49 zones randomly allocated to receive the Newhints intervention, a mean of 407 (SD 18) community-based surveillance volunteers undertook home visits for 7848 pregnant women who gave birth to 7786 live babies in 2009. Annual economic cost of implementation was US$203 998, or $0·53 per person. In the base-case analysis, the Newhints intervention cost a mean of $10 343 (95% CI 2963 to -7674) per newborn life saved, or $352 (95% CI 104 to -268) per discounted life-year saved, and had a 72% chance of being highly cost effective with respect to Ghana's 2009 gross domestic product per person. Key determinants of cost-effectiveness were the discount rate, protective effectiveness, baseline neonatal mortality rate, and implementation costs. In the scenarios modelled with the meta-analysis results, the ICER increased from $127 per life-year saved at a neonatal mortality rate of 60 deaths per 1000 livebirths, to $379 per life-year saved at a rate of 20 deaths per 1000 livebirths. The strategy had at least a 99% probability of being highly cost effective for lower-middle-income countries in all neonatal mortality rate scenarios modelled, and at least a 95% probability of being highly cost effective for low-income countries at neonatal mortality rates of 30 or more deaths per 1000 livebirths. Our findings show that the seemingly modest mortality reductions achieved by a newborn home-visit strategy might in fact be cost effective. In Ghana, such strategies are also likely to be affordable. Our findings support recommendations from WHO and UNICEF that low-income and middle-income countries implement newborn home visits. The Bill & Melinda Gates Foundation, UK Department for International Development, WHO.


Structure and conformational plasticity of the U6 small nuclear ribonucleoprotein core.
U6 small nuclear RNA (snRNA) is a key component of the active site of the spliceosome, a large ribonucleoprotein complex that catalyzes the splicing of precursor messenger RNA. Prior to its incorporation into the spliceosome, U6 is bound by the protein Prp24, which facilitates unwinding of the U6 internal stem-loop (ISL) so that it can pair with U4 snRNA. A previously reported crystal structure of the `core' of the U6 small nuclear ribonucleoprotein (snRNP) contained an ISL-stabilized A62G mutant of U6 bound to all four RNA-recognition motif (RRM) domains of Prp24 [Montemayor et al. (2014), Nature Struct. Mol. Biol. 21, 544-551]. The structure revealed a novel topology containing interlocked rings of protein and RNA that was not predicted by prior biochemical and genetic data. Here, the crystal structure of the U6 snRNP core with a wild-type ISL is reported. This complex crystallized in a new space group, apparently owing in part to the presence of an intramolecular cross-link in RRM1 that was not observed in the previously reported U6-A62G structure. The structure exhibits the same protein-RNA interface and maintains the unique interlocked topology. However, the orientation of the wild-type ISL is altered relative to the A62G mutant structure, suggesting inherent structural dynamics that may facilitate its pairing with U4. Consistent with their similar architectures in the crystalline state, the wild-type and A62G variants of U6 exhibit similar Prp24-binding affinities and electrophoretic mobilities when analyzed by gel-shift assay.


Effects of calcium supplementation on clinical fracture and bone structure: results of a 5-year, double-blind, placebo-controlled trial in elderly women.
Increased dietary calcium intake has been proposed as a population-based public health intervention to prevent osteoporotic fractures. We have examined whether calcium supplementation decreases clinical fracture risk in elderly women and its mechanism of action. Five-year, double-blind, placebo-controlled study of 1460 women recruited from the population and older than 70 years (mean age, 75 years) who were randomized to receive calcium carbonate, 600 mg twice per day, or identical placebo. The primary end points included clinical incident osteoporotic fractures, vertebral deformity, and adverse events ascertained in 5 years. Bone structure was also measured using dual x-ray absorptiometry of the hip and whole body, quantitative ultrasonography of the heel, and peripheral quantitative computed tomography of the distal radius. Among our patients, 16.1% sustained 1 or more clinical osteoporotic fractures. In the intention-to-treat analysis, calcium supplementation did not significantly reduce fracture risk (hazard ratio, 0.87; 95% confidence interval, 0.67-1.12). However, 830 patients (56.8%) who took 80% or more of their tablets (calcium or placebo) per year had reduced fracture incidence in the calcium compared with the placebo groups (10.2% vs 15.4%; hazard ratio, 0.66; 95% confidence interval, 0.45-0.97). Calcium-treated patients had improved quantitative ultrasonography findings of the heel, femoral neck and whole-body dual x-ray absorptiometry data, and bone strength compared with placebo-treated patients. Of the 92 000 adverse events recorded, constipation was the only event increased by the treatment (calcium group, 13.4%; placebo group, 9.1%). Supplementation with calcium carbonate tablets supplying 1200 mg/d is ineffective as a public health intervention in preventing clinical fractures in the ambulatory elderly population owing to poor long-term compliance, but it is effective in those patients who are compliant.


Kaposi sarcoma incidence and survival among HIV-infected homosexual men after HIV seroconversion.
Despite the success of combination antiretroviral therapy (cART) in reducing the incidence of Kaposi sarcoma, HIV-infected individuals who have responded to treatment continue to be diagnosed with Kaposi sarcoma. We examine factors associated with the incidence of Kaposi sarcoma among cART-treated HIV-infected homosexual men and changes in their survival after its diagnosis over calendar time. Data were from HIV-infected homosexual men with well-estimated dates of HIV seroconversion (ie, change in status from being HIV negative to having HIV antibodies detected). Incidence of Kaposi sarcoma was calculated. We used Kaplan-Meier methods to determine survival after Kaposi sarcoma diagnosis in three calendar periods: before 1996, 1996-2000, and 2001-2006. Poisson models were used to examine the effect of risk factors such as current and nadir CD4 cell count (ie, the lowest CD4 cell count ever recorded for a person), duration of infection, and age at diagnosis for Kaposi sarcoma incidence in cART-treated men. All statistical tests were two-sided. Among the 9473 men, 555 were diagnosed with Kaposi sarcoma in the period 1986-2006, of whom 319 died. The percentage surviving 24 months after Kaposi sarcoma diagnosis rose statistically significantly during the study period from 35% (95% confidence interval [CI] = 29% to 42%) before 1996 to 84% (95% CI = 76% to 90%) in 1996-2000 and to 81% (95% CI = 70% to 88%) in 2001-2006 (P < .001). Seventy men were diagnosed with Kaposi sarcoma after starting cART. Current (ie, within 6 months) CD4 cell count was associated with incidence of Kaposi sarcoma among cART-treated men (rate ratios [RRs] = 18.91, 95% CI = 8.50 to 42.09, for CD4 level category <200 cells per cubic millimeter; RR = 3.55, 95% CI = 1.40 to 9.00, for 200-349 cells per cubic millimeter; and RR = 4.11, 95% CI = 1.74 to 9.70, for 350-499 cells per cubic millimeter; all compared with > or = 500 cells per cubic millimeter). After adjustment for current CD4 cell count, HIV infection duration, age, or nadir CD4 cell count was not associated with Kaposi sarcoma incidence. Among cART-treated HIV-infected homosexual men, current CD4 cell count was the factor most strongly associated with the incidence of Kaposi sarcoma. Survival estimates after Kaposi sarcoma diagnosis have improved over time.


2D IR spectra of cyanide in water investigated by molecular dynamics simulations.
Using classical molecular dynamics simulations, the 2D infrared (IR) spectroscopy of CN(-) solvated in D2O is investigated. Depending on the force field parametrizations, most of which are based on multipolar interactions for the CN(-) molecule, the frequency-frequency correlation function and observables computed from it differ. Most notably, models based on multipoles for CN(-) and TIP3P for water yield quantitatively correct results when compared with experiments. Furthermore, the recent finding that T1 times are sensitive to the van der Waals ranges on the CN(-) is confirmed in the present study. For the linear IR spectrum, the best model reproduces the full widths at half maximum almost quantitatively (13.0 cm(-1) vs. 14.9 cm(-1)) if the rotational contribution to the linewidth is included. Without the rotational contribution, the lines are too narrow by about a factor of two, which agrees with Raman and IR experiments. The computed and experimental tilt angles (or nodal slopes) α as a function of the 2D IR waiting time compare favorably with the measured ones and the frequency fluctuation correlation function is invariably found to contain three time scales: a sub-ps, 1 ps, and one on the 10-ps time scale. These time scales are discussed in terms of the structural dynamics of the surrounding solvent and it is found that the longest time scale (≈10 ps) most likely corresponds to solvent exchange between the first and second solvation shell, in agreement with interpretations from nuclear magnetic resonance measurements.


First Report of a Leaf Spot on Hazel Leaves Caused by Phyllosticta coryli in Liaoning Province of China.
Hazel (Corylus heterophylla Fischl) is an important nut tree grown in China, especially in Liaoning Province, and is rich in nutritional and medicinal values. In August 2011, leaf spotting was observed on hybrid hazel (Dawei) leaves in Paotai Town, Wafangdian County of Liaoning Province. By August 2012, the disease had spread to Zhangdang Town, Fushun County. Symptoms initially appeared on both sides of leaves as pinpoint brown spots, which enlarged and developed into regular, dark brown lesions, 3 to 9 mm in diameter. The lesions were lighter in color in the center compared to the margin. To identify the pathogen, leaf pieces (3 to 5 mm) taken from the margins, including both symptomatic and healthy portions of leaf tissue, were surface-disinfected first in 75% ethanol for 5 s, next in 0.1% aqueous mercuric chloride for 50 s, and then rinsed with sterilized water three times. Leaf pieces were incubated on potato dextrose agar (PDA) at 25°C for 14 days in darkness. Single spore isolates were obtained from individual conidia. For studies of microscopic morphology, isolates were grown on synthetic nutrient agar (SNA) in slide cultures. Colonies grew up to 45 to 48 mm in diameter on PDA after 14 days. Pycnidia appeared on the colonies after 12 days. Conidiophores were short. Pycnidia were dark brown, subglobose, and 150 to 205 μm in diameter. Conidia were unicellular, colorless, ovoid to oval, and from 2.4 to 4.5 × 1.6 to 2.4 μm. On the basis of these morphological characteristics, the isolates were tentatively identified as Phyllosticta coryli Westend (2). The rDNA internal transcribed spacer (ITS) region was amplified using primers ITS1 and ITS4 and sequenced (GenBank Accession No. KC196068). The 490-bp amplicons had 100% identity to an undescribed Phyllosticta species isolated from Cornus macrophylla in Gansu, Tianshui, China (AB470897). On the basis of morphological characteristics and nucleotide homology, the isolate was tentatively identified as P. coryli. Koch's postulates were fulfilled in the growth chamber on hazelnut leaves inoculated with P. coryli conidial suspensions (107 conidia ml-1). Eight inoculated 1-year-old seedlings (Dawei) were incubated under moist conditions for 8 to 10 days at 25°C. All leaf spots that developed on inoculated leaves were similar in appearance to those observed on diseased hazel leaves in the field. P. coryli was recovered from lesions and its identity was confirmed by morphological characteristics. P. coryli was first reported as a pathogen of hazel leaves in Bull of Belgium (2). In China, P. coryli was first reported on Corylus heterophylla Fisch. in Jilin Province (1). To our knowledge, this is the first report of P. coryli causing leaf spot on hybrid hazel in Liaoning Province of China. The outbreak and spread of this disease may decrease the yield of hazelnut in northern regions of China. More studies are needed on control strategies, including the possible resistance of hazel cultivars to P. coryli. References: (1) Y. Li et al. J. Shenyang Agric. Univ. 25:153, 1994. (2) P. A. Saccardo. Sylloge Fungorum Vol. III, page 31, 1884.


Effects of hypothermia on brain glucose metabolism in acute liver failure: a H/C-nuclear magnetic resonance study.
Mild hypothermia has a protective effect on brain edema and encephalopathy in both experimental and human acute liver failure. The goals of the present study were to examine the effects of mild hypothermia (35 degrees C) on brain metabolic pathways using combined (1)H and (13)C-Nuclear Magnetic Resonance (NMR) spectroscopy, a technique which allows the study not only of metabolite concentrations but also their de novo synthesis via cell-specific pathways in the brain. (1)H and (13)C NMR spectroscopy using [1-(13)C] glucose was performed on extracts of frontal cortex obtained from groups of rats with acute liver failure induced by hepatic devascularization whose body temperature was maintained either at 37 degrees C (normothermic) or 35 degrees C (hypothermic), and appropriate sham-operated controls. At coma stages of encephalopathy in the normothermic acute liver failure animals, glutamine concentrations in frontal cortex increased 3.5-fold compared to sham-operated controls (P < 0.001). Comparable increases of brain glutamine were observed in hypothermic animals despite the absence of severe encephalopathy (coma). Brain glutamate and aspartate concentrations were respectively decreased to 60.9% +/- 7.7% and 42.2% +/- 5.9% (P < 0.01) in normothermic animals with acute liver failure compared to control and were restored to normal values by mild hypothermia. Concentrations of lactate and alanine in frontal cortex were increased to 169.2% +/- 15.6% and 267.3% +/- 34.0% (P < 0.01) respectively in normothermic rats compared to controls. Furthermore, de novo synthesis of lactate and alanine increased to 446.5% +/- 48.7% and 707.9% +/- 65.7% (P < 0.001), of control respectively, resulting in increased fractional (13)C-enrichments in these cytosolic metabolites. Again, these changes of lactate and alanine concentrations were prevented by mild hypothermia. Mild hypothermia (35 degrees C) prevents the encephalopathy and brain edema resulting from hepatic devascularization, selectively normalizes lactate and alanine synthesis from glucose, and prevents the impairment of oxidative metabolism associated with this model of ALF, but has no significant effect on brain glutamine. These findings suggest that a deficit in brain glucose metabolism rather than glutamine accumulation is the major cause of the cerebral complications of acute liver failure.


Characterization of novel antimicrobial peptides from the skins of frogs of the Rana esculenta complex.
Rana esculenta is a hybridogenetic hybrid between Rana ridibunda and Rana lessonae and so is best considered as a complex of interbreeding species rather than a discrete single species. In this study, antimicrobial peptides were isolated from a pooled extract of the skins of specimens of the R. esculenta complex collected in the wild. In addition to several peptides belonging to the brevinin and esculentin families that have been previously isolated from skin secretions of a single specimen of R. esculenta, three newly described members of the brevinin-2 family (brevinin-2Ei, brevinin-2Ej, and brevinin-2Ek) and one member of the temporin family (temporin-1Ec) were purified and characterized. In addition, three structurally related peptides with no sequence similarity with antimicrobial peptides isolated from other species of ranid frogs, that potently and selectively inhibit the growth of the Gram-positive bacterium Escherichia coli (minimal inhibitory concentration (MIC<5 microM)), were identified. These peptides show limited amino acid sequence similarity to the homologous exon gene products that encode the N-terminal flanking peptides of preprocaerulein, preproxenopsin, and preprolevitide and so have been termed caerulein precursor-related fragments (CPRF-Ea, CPRF-Eb, and CPRF-Ec). The data suggest that there may be considerable polymorphism among specimens from different populations of the R. esculenta complex. It is proposed that the distribution and amino acid sequences of skin antimicrobial peptides may be useful markers for taxonomic classification of particular sub-populations and for an understanding of phylogenetic interrelationships.


HEG1, BAP1, and MTAP are useful in cytologic diagnosis of malignant mesothelioma with effusion.
The specificity and sensitivity of HEG1 for malignant mesothelioma (MM) is high. The use of BAP1/MTAP immunohistochemistry (IHC) is recommended to separate benign and malignant mesothelial proliferations. We determined how ancillary techniques can be used for the cytological diagnosis of MM with effusion. Cell blocks from effusions from cases with MM, reactive mesothelial cells (RMCs), and carcinomas were analyzed by IHC with HEG1, BAP1, and MTAP and with homozygous deletion (HD) of CDKN2A by fluorescence in situ hybridization. Staining scores were calculated for IHC by adding the number of categories for the staining intensity and the staining extension. HEG1 was positive in all (41/41) MMs, but negative in carcinomas, except for ovarian carcinomas. Overall 76.9% (20/26) of RMCs and 28.6% (6/21) of ovarian carcinomas expressed HEG1. BAP1 loss was found in 71.1% of MMs, but none was found in RMCs. MTAP loss was found in 76.2% of MMs, but none was found in RMCs. 73.9% of MMs harbored HD of CDKN2A. There was concordance between loss of MTAP and HD of CDKN2A in 95% of MMs. HEG1 is a good marker for mesothelial differentiation in effusion cytology. HD of CDKN2A is frequently observed in cell blocks from effusions of MMs, and MTAP IHC may act as a surrogate for HD of CDKN2A. Cell block analysis is recommended for effusions of unknown origins with the following methods: IHC with HEG1 and claudin 4 to validate the mesothelial origin, followed by BAP1 and MTAP IHC to confirm malignancy.


2022 American College of Rheumatology/EULAR Classification Criteria for Takayasu Arteritis.
To develop and validate new classification criteria for Takayasu arteritis (TAK). Patients with vasculitis or comparator diseases were recruited into an international cohort. The study proceeded in 6 phases: 1) identification of candidate criteria items, 2) collection of candidate items present at diagnosis, 3) expert panel review of cases, 4) data-driven reduction of candidate items, 5) derivation of a points-based classification score in a development data set, and 6) validation in an independent data set. The development data set consisted of 316 cases of TAK and 323 comparators. The validation data set consisted of an additional 146 cases of TAK and 127 comparators. Age ≤60 years at diagnosis and imaging evidence of large-vessel vasculitis were absolute requirements to classify a patient as having TAK. The final criteria items and weights were as follows: female sex (+1), angina (+2), limb claudication (+2), arterial bruit (+2), reduced upper extremity pulse (+2), reduced pulse or tenderness of a carotid artery (+2), blood pressure difference between arms of ≥20 mm Hg (+1), number of affected arterial territories (+1 to +3), paired artery involvement (+1), and abdominal aorta plus renal or mesenteric involvement (+3). A patient could be classified as having TAK with a cumulative score of ≥5 points. When these criteria were tested in the validation data set, the model area under the curve was 0.97 (95% confidence interval [95% CI] 0.94-0.99) with a sensitivity of 93.8% (95% CI 88.6-97.1%) and specificity of 99.2% (95% CI 96.7-100.0%). The 2022 American College of Rheumatology/EULAR classification criteria for TAK are now validated for use in research.


Expression changes in arrestin β 1 and genetic variation in catechol-O-methyltransferase are biomarkers for the response to morphine treatment in cancer patients.
Genetic differences in individuals with regard to opioid-receptor signaling create clinical difficulties for opioid treatment; consequently, useful pharmacodynamic and predictive biomarkers are needed. In this prospective study, we studied gene expression changes in peripheral blood leukocytes using a microarray and real-time RT-PCR analysis to identify pharmacodynamic biomarkers for monitoring the effect of morphine in a cohort of opioid-treatment-naïve cancer patients. We also examined genetic variations in opioid receptor mu 1 (OPRM1, 118A→G) and catechol-O-methyltransferase (COMT, 472G→A) to evaluate predictive biomarkers of the treatment outcome of morphine. The plasma concentration of morphine was measured using a liquid chromatography-tandem mass spectrometry method. Microarray analysis revealed that the mRNA expression levels of arrestin β 1 (ARRB1) were significantly down-regulated by morphine treatment. Real-time RT-PCR analysis against independent samples confirmed the results (P=0.003) and changes during treatment were negatively correlated with the plasma morphine concentration (R=-0.42). No correlation was observed between the genotype of OPRM1 and morphine treatment; however, the plasma concentration of morphine and the required dose of morphine were significantly lower for the A/A genotype of COMT (vs. A/G+G/G, P=0.008 and 0.03). We found that changes in the expression of ARRB1 may be a novel pharmacodynamic biomarker and the COMT 472G→A genotype may be a predictive biomarker of the response to morphine treatment.


Facile electrospinning formation of carbon-confined metal oxide cube-in-tube nanostructures for stable lithium storage.
A unique carbon-confined metal oxide cube-in-tube nanostructure is synthesized by a facile precursor-modified electrospinning method with subsequent pyrolysis. This nanostructure has a partly graphitized carbon layer with manganese oxide nanoparticles embedded as the tube and amorphous CoSnO3 hollow cubes uniformly distributed inside the tube. As a lithium-ion battery anode, this architecture exhibits a high reversible discharge capacity and rate capability.


Diseases that precede disability among latter-stage elderly individuals in Japan.
Understanding causes of disability among elderly individuals is an important public health issue, particularly because of the increasing rate of disabled elderly individuals and the social costs in a rapidly aging society. Accordingly, we aimed to describe the diseases that precede disability and investigate the types of diseases that are related to severe disability among Japanese elderly individuals aged over 75 years. Using claim data from the latter-stage elderly healthcare system and long-term care insurance system, we identified 76,265 elderly individuals over 75 years old who did not qualify as disabled on April 1, 2011. Among them, 3,715 elderly individuals who had been newly qualified as disabled between April 1, 2011 and March 31, 2012 were selected. Disease codes from the medical claim data in the 6 months prior to disability were collected. All descriptions were developed separately for six groups divided by gender and disability level (low, middle, and high). The results of the ordinal logistic analysis including sex and age revealed that men tended to have significantly higher levels of disability (β = 0.417, p < 0.001) than women. Cerebrovascular disorder (CVD) was the most common disease in almost all age and disability level groups. In low-level disability groups, cancer in men (12.8%) and arthropathy and fracture in women (11.9% and 13.5%, respectively) were as common as cerebrovascular disorder (12.2% and 9.7%, in men and women, respectively). Stroke was the most common disease for all genders and disability levels. The diseases preceding low-level disability differed by gender. This study demonstrated the need to consider arthropathy and fracture as well as CVD in order to prevent disability.