A standardised tool for assessing needs in forensic psychiatric population: clinical validation of the Italian CANFOR, staff version.
The Camberwell Assessment of Need - Forensic Version (CANFOR) is a standardised assessment tool specifically designed to assess needs for care in forensic psychiatric populations. The original English version of the instrument has shown good psychometric properties. The aim of this study was to validate the Italian version of the CANFOR-staff tool. After translation and back-translation, the Italian CANFOR tool was administered to a sample of 50 forensic psychiatric patients. Convergent validity was tested using the Brief Psychiatric Rating Scale (BPRS) and the Global Assessment of Functioning (GAF) by applying Kendall's tau-b. Inter-rater and test-retest reliabilities were measured by ICCs for need dimensions (total and unmet) and Cohen's kappa coefficients for individual need items. Regarding convergent validity, a higher number of needs (total and unmet) were associated with more severe psychiatric symptoms (BPRS). Higher numbers of unmet needs were also associated with lower levels of global functioning (GAF). ICCs for total and unmet needs scores indicated a good level of agreement for inter-rater reliability and a very good level for test-retest, respectively. Regarding the specific items, inter-rater Cohen's kappa was high (moderate to very good agreement) for 18 items in relation to the presence of a need and for 15 items in the rating of an unmet need, whereas Cohen's kappa for test-retest reliability was very high for all the items in the presence of a need and high for 18 of the unmet need domains. The Italian version of CANFOR has adequate psychometric properties. It can be considered a promising instrument for the assessment of needs of forensic psychiatric patients.


The expression and distribution of dense granule proteins in the enteric (Coccidian) forms of Toxoplasma gondii in the small intestine of the cat.
The expression and distribution of dense granule proteins in the enteric (coccidian) forms of Toxoplasma gondii in the small intestine of the cat. Experimental Parasitology 91, 203-211. The expression and location of the dense granule proteins (GRA1-6 and NTPase) in the merozoite and during asexual and sexual development of Toxoplasma gondii in the small intestine of the cat (definitive host) was examined by immuno-light and electron microscopy. This was compared with that of tachyzoites and bradyzoites present in the intermediate host. It was found that the merozoite contained the characteristic apical organelles plus a few large dense granules. By immunocytochemistry, dense granules in merozoites were negative for GRA proteins 1 to 6 in contrast to both tachyzoites and bradyzoites in which dense granules were positive for all six proteins. The GRA proteins were associated with the parasitophorous vacuole (PV) during tachyzoite and bradyzoite development but were absent from the PV of the enteric stages. However, the merozoite dense granules were positive for NTPase, which was similar to the tachyzoite while this antigen was down regulated in the bradyzoite. The apparent release of the NTPases into the PV formed by merozoites was also similar to that described for the tachyzoite, possibly reflecting the relative metabolic activity of the various stages. This study shows that the majority of GRA proteins have a similar stage-specific expression, which is independent of NTPases expression. These observations are consistent with T. gondii having a different host parasite relationship in the enteric forms, which does not involve the GRA proteins 1-6.


Throwing new light on lung cancer pathogenesis: updates on three recent topics.
Lung cancers have become the leading cause of cancer deaths in Japan, claiming more than 55 000 lives annually. Unfortunately, substantial improvement in terms of cure rates has not been achieved over the last two decades, although during the same period of time in-depth basic knowledge of the molecular mechanisms, which underlies carcinogenesis and progression of this deadly group of neoplasms, has accumulated at an amazing pace. It has consequently become evident that they have many shared but also distinct features, when comparisons are made not only with other common epithelial cancers of adults, such as colon cancer, but also within the various histologic types of lung cancers themselves. This review article provides an up-date on cutting-edge research into the following three different topics, from which important new insights have been obtained. The first concerns genetic instability, especially chromosome instability, and checkpoint failure in lung cancers. Second, we deal with EGFR mutations, which shows revealing specificities in various aspects. Finally, advances in the expression profiling analysis of both transcriptomes and proteomes of lung cancers are summarized.


Assessing the scaffold diversity of screening libraries.
Medicinal chemists have traditionally realized assessments of chemical diversity and subsequent compound acquisition, although a recent study suggests that experts are usually inconsistent in reviewing large data sets. To analyze the scaffold diversity of commercially available screening collections, we have developed a general workflow aimed at (1) identifying druglike compounds, (2) clustering them by maximum common substructures (scaffolds), (3) measuring the scaffold diversity encoded by each screening collection independently of its size, and finally (4) merging all common substructures in a nonredundant scaffold library that can easily be browsed by structural and topological queries. Starting from 2.4 million compounds out of 12 commercial sources, four categories of libraries could be identified: large- and medium-sized combinatorial libraries (low scaffold diversity), diverse libraries (medium diversity, medium size), and highly diverse libraries (high diversity, low size). The chemical space covered by the scaffold library can be searched to prioritize scaffold-focused libraries.


Class I major histocompatibility complex-restricted cytotoxic T lymphocytes specific for Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell lines against which they were raised.
We have raised CD8+ cytotoxic T lymphocytes (CTL) from three Epstein-Barr virus-seropositive donors by incubating peripheral blood lymphocytes with irradiated autologous B95.8-strain EBV-transformed B lymphoblastoid cells (LCL). However, to detect lysis in a standard 51Cr release assay of the LCL against which these CTL were raised, superinfection with recombinant vaccinia expressing the appropriate EBV protein or incubation with the peptide epitope was necessary. The untreated LCL were not lysed, even though Western blotting demonstrated that they expressed the EBV antigens containing the CTL epitopes. We have found CTL of this phenotype that are restricted by human leukocyte antigen-A2, -A3, -B7, or -B39, and which recognize the EBV latent proteins, EBV nuclear antigen (EBNA)-3A, EBNA-3C, or terminal protein. During these experiments, we identified a new human leukocyte antigen-A3-restricted EBNA-3A epitope, residues 603-611, RLRAEAGVK. We raised a spontaneous LCL, transformed by endogenous EBV, from one donor, but this was also not lysed. For at least one of the epitopes, CTL from another donor lysed the LCL without superinfection or addition of peptides. We conclude that the CTL were unable to achieve a high enough avidity of interaction with untreated LCL to trigger effector function, although the LCL were able to stimulate them to grow in vitro for up to 4 mo. To assess whether a small percentage of the LCL might possess a higher antigen density, we used an assay of tumor necrosis factor release from a CTL clone, which was able to detect antigen-bearing cells representing only 1% of a stimulating LCL population. Nevertheless, the untreated autologous LCL line failed to stimulate tumor necrosis factor release.


Return to performance following severe ankle, knee, and hip injuries in National Basketball Association players.
The purpose of this study was to compare basketball performance markers 1 y prior to initial severe lower extremity injury, including ankle, knee, and hip injuries, to 1 and 2 y following injury during the regular National Basketball Association (NBA) season. Publicly available data were extracted through a reproducible extraction computed programmed process. Eligible participants were NBA players with at least three seasons played between 2008 and 2019, with a time-loss injury reported during the study period. Basketball performance was evaluated for season minutes, points, and rebounds. Prevalence of return to performance and linear regressions were calculated. A total of 285 athletes sustained a severe lower extremity injury. A total of 196 (69%) played for 1 y and 130 (45%) played for 2 y following the injury. A total of 58 (30%) players participated in a similar number of games and 57 (29%) scored similar points 1 y following injury. A total of 48 (37%) participated in a similar number of games and 55 (42%) scored a similar number of points 2 y following injury. Fewer than half of basketball players who suffered a severe lower extremity injury were participating at the NBA level 2 y following injury, with similar findings for groin/hip/thigh, knee, and ankle injuries. Fewer than half of players were performing at previous preinjury levels 2 y following injury. Suffering a severe lower extremity injury may be a prognostic factor that can assist sports medicine professionals to educate and set performance expectations for NBA players.


Biogenic emissions of isoprenoids and NO in China and comparison to anthropogenic emissions.
In this study, a regional dynamical model (WRF) is used to drive biogenic emission models to calculate high resolution (10x10 km) biogenic emissions of isoprene (C(5)H(8)), monoterpenes (C(10)H(16)), and nitric oxide (NO) in China. This high resolution biogenic inventory will be available for the community to study the effect of biogenic emissions on photochemical oxidants in China. The biogenic emissions are compared to anthropogenic emissions to gain insight on the potential impact of the biogenic emissions on tropospheric chemistry, especially ozone production in this region. The results show that the biogenic emissions in China exhibit strongly diurnal, seasonal, and spatial variations. The isoprenoid (including both isoprene and monoterpenes) emissions are closely correlated to tree density and strongly vary with season and local time. During winter (January), the biogenic isoprenoid emissions are the lowest, resulting from lower temperature and solar radiation, and highest in summer (July) due to higher temperature and solar radiation. The biogenic NO emissions are also higher during summer and lower during winter, but the magnitude of the seasonal variation is smaller than the emissions of isoprene and monoterpenes. The biogenic emissions of NO are widely spread out in the northern, eastern, and southern China regions, where high-density agricultural soil lands are located. Both biogenic NO and isoprenoid emissions are very small in western China. The calculated total biogenic emission budget is smaller than the total anthropogenic VOC emission budget in China. The biogenic isoprenoid and anthropogenic VOC emissions are 10.9 and 15.1 Tg year(-1), respectively. The total biogenic and anthropogenic emissions of NO are 5.9 and 11.5 Tg(NO) year(-1), respectively. The study shows that in central eastern China, the estimated biogenic emissions of isoprenoids are very small, and the anthropogenic emissions of VOCs are dominant in this region. However, in northeastern and southern China, there are relatively large biogenic emissions of isoprenoids, leading to an important impact on the ozone production in these regions. Furthermore, the emissions of isoprenoids are highest during summer and noontime, which correlates to the peak of ozone production period. For example, the ratio between summer and winter for the emissions of isoprenoids is about 15 in China. As a result, the biogenic emissions of isoprenoids are significantly larger than the anthropogenic emissions of VOCs in China during daytime in summer. Biogenic NO emissions are mostly produced by agricultural soils which co-exist with large populations and human activity. As a result, the biogenic emissions of NO are mostly overlapped with the anthropogenic emissions of NO, leading to the enhancement in NO concentrations in the high anthropogenic NO emission regions. Finally, the future emissions of isoprene and monoterpenes over China are estimated. The results show that the future biogenic emissions may increase significantly due to land cover changes in central eastern China, which could have a very important impact on ozone formation in this region. However, these estimates are highly uncertain and are presented as a potential scenario to show the importance of possible changes of biogenic emissions in China.


Fine-particulate air pollution and life expectancy in the United States.
Exposure to fine-particulate air pollution has been associated with increased morbidity and mortality, suggesting that sustained reductions in pollution exposure should result in improved life expectancy. This study directly evaluated the changes in life expectancy associated with differential changes in fine particulate air pollution that occurred in the United States during the 1980s and 1990s. We compiled data on life expectancy, socioeconomic status, and demographic characteristics for 211 county units in the 51 U.S. metropolitan areas with matching data on fine-particulate air pollution for the late 1970s and early 1980s and the late 1990s and early 2000s. Regression models were used to estimate the association between reductions in pollution and changes in life expectancy, with adjustment for changes in socioeconomic and demographic variables and in proxy indicators for the prevalence of cigarette smoking. A decrease of 10 microg per cubic meter in the concentration of fine particulate matter was associated with an estimated increase in mean (+/-SE) life expectancy of 0.61+/-0.20 year (P=0.004). The estimated effect of reduced exposure to pollution on life expectancy was not highly sensitive to adjustment for changes in socioeconomic, demographic, or proxy variables for the prevalence of smoking or to the restriction of observations to relatively large counties. Reductions in air pollution accounted for as much as 15% of the overall increase in life expectancy in the study areas. A reduction in exposure to ambient fine-particulate air pollution contributed to significant and measurable improvements in life expectancy in the United States.


The quest for Y-chromosomal markers - methodological strategies for mammalian non-model organisms.
Tracing maternal and paternal lineages independently to explore breeding systems and dispersal strategies in natural populations has been high on the wish-list of evolutionary biologists. As males are the heterogametic sex in mammals, such sex-specific patterns can be indirectly observed when Y chromosome polymorphism is combined with mitochondrial sequence information. Over the past decade, Y-chromosomal markers applied to human populations have revealed remarkable differences in the demographic history and behaviour between the sexes. However, with a few exceptions, genetic data tracing the paternal line are lacking in most other mammalian species. This deficit can be attributed to the difficulty of developing Y-specific genetic markers in non-model organisms and the general low levels of polymorphisms observed on the Y chromosome. Here, we present an overview of the currently employed strategies for developing paternal markers in mammals. Moreover, we review the practical feasibility and requirements of various methodological strategies and highlight their future prospects when combined with new molecular techniques such as next generation sequencing.


Role of epidermis-type lipoxygenases for skin barrier function and adipocyte differentiation.
12R-lipoxygenase (12R-LOX) and epidermis-type LOX-3 (eLOX-3) are novel members of the multigene family of mammalian LOX. A considerable gap exists between the identification of these enzymes and their biologic function. Here, we present evidence that 12R-LOX and eLOX-3, acting in sequence, and eLOX-3 in combination with another, not yet identified LOX are critically involved in terminal differentiation of keratinocytes and adipocytes, respectively. Mutational inactivation of 12R-LOX and/or eLOX-3 has been found to be associated with development of an inherited ichthyosiform skin disorder in humans and genetic ablation of 12R-LOX causes a severe impairment of the epidermal lipid barrier in mice leading to post-natal death of the animals. In preadipocytes, a LOX-dependent PPARgamma activating ligand is released into the cell supernatant early upon induction of differentiation and available evidence indicates that this ligand is an eLOX-3-derived product. In accordance with this data is the observation that forced expression of eLOX-3 enhances adipocyte differentiation.


Benzo(a)pyrene disrupts mouse preimplantation embryo development.
To determine the effects of Benzo(a)pyrene (BaP) on the development of early preimplantation embryo by exposure to physiologic concentrations of BaP based on a previous report in human ovarian follicular fluid and serum. Zygotes were cultured in 5 nM or 50 nM BaP and then examined for development efficiency, embryo quality, and DNA damage. In addition, embryonic stem cells (ESCs) were used as a model to test the toxic effects of BaP on inner cell mass (ICM) of blastocysts. Laboratory. CD1 mice. Mouse zygotes and ESCs were cultured in medium with 5 nM or 50 nM BaP. The percentage (rate) of blastocyst development, reactive oxygen species level, and quality of embryos assessed by total cell number, cell apoptosis, Oct4- and Nanog-positive cell ratio, and DNA damage on genomic and telomeric DNA were compared between dimethyl sulfoxide control and BaP treatments. The BaP-treated zygotes exhibited significantly higher reactive oxygen species activity, which might lead to more cell apoptosis, low ratio of Nanog- or Oct4-positive ICM cells, and increasing DNA damage in both genomic and telomeric DNA in blastocysts. By using mouse ESCs derived from ICM cells as a model, we showed that pluripotent cells might also show serious DNA damage after a brief exposure to BaP. Our data show that BaP could seriously disrupt cell growth and genomic DNA stability and increase cell apoptosis in mouse preimplantation embryo development.


Suppression of G-protein-coupled receptor kinase 3 expression is a feature of classical GBM that is required for maximal growth.
G-protein-coupled receptor kinases (GRK) regulate the function of G-protein-coupled receptors (GPCR). Previously, we found that GPCR (CXCR4)-mediated astrocytoma growth was dependent upon abnormally sustained CXCR4 signaling and was correlated with decreased GRK-mediated receptor phosphorylation. As CXCR4 has also been implicated in the stimulation of high-grade glioma growth, we sought to determine whether dysregulation of GRK expression and/or function might also be present in high-grade gliomas. In an analysis of data from The Cancer Genome Atlas, we found that GRK3 expression is frequently decreased in glioblastoma (GBM) of the classical subtype, which possesses signature amplification or mutational activation of the epidermal growth factor (EGF) receptor. We tested the correlation between GRK3 expression and GBM subtypes, as well as the relationship between the activation of the EGF and other growth factor receptor pathways and GRK expression. In analyses of primary GBM tissue and RNA specimens, we found that GRK3 expression is correlated with established criteria for GBM subtyping including expression of EGF receptor, platelet-derived growth factor receptor (PDGFR)α, NF1, PTEN, CDKN2A, and neurofilament. We also found that established drivers of gliomagenesis, the EGF, PDGF, and TGF-β pathways, all regulate GRK expression. Coculture experiments, designed to mimic critical interactions between tumor and brain microvascular endothelial cells, showed that specifically increasing GRK3 expression reduced the trophic effect of endothelial cells on tumor cells. Together, these experiments show that GRK3 is a negative regulator of cell growth whose expression is preferentially reduced in GBM of the classical subtype as a consequence of activity in primary gliomagenic pathways.


Quality assurance of drug therapy in hospitals: patient serum creatinine values used by ward pharmacists in checking dosage regimens.
A programme for the systematic adjustment of drug dosage in hospital patients with impairment of renal function is described. This programme involves the efficient identification of patients with elevated serum levels of creatinine, the checking of dosage regimens for these patients in terms of known pharmacokinetic data, and advising of medical officers when dosage regimens are in excess of those recommended, or where contraindicated drugs have been prescribed. The procedure, which has been shown to be both workable and effective in a large teaching hospital over a period of more than two years, serves as a model for quality assurance of an aspect of drug treatment with existing staff and with no additional capital costs.


Endoproteolysis of the ER stress transducer ATF6 in the presence of functionally inactive presenilins.
Presenilin (PS) proteins facilitate endoproteolysis of selected type I transmembrane proteins such as the Alzheimer's disease (AD) associated beta-Amyloid precursor protein (beta APP) and Notch. beta APP is cleaved within its transmembrane domain by an aspartyl protease activity termed gamma-secretase, which may be identical with PS1 and PS2. Notch also undergoes a PS-dependent intramembraneous proteolysis. A similar gamma-secretase-like cleavage may also occur with IRE1 and ATF6, two signaling molecules of the unfolded protein response (UPR) that may require PSs for their activation. Here, we have analyzed whether ATF6 cleavage requires a PS-dependent gamma-secretase activity and whether inhibition of gamma-secretase activity would affect the UPR. Endoproteolysis of ATF6 was observed in the presence of the highly potent gamma-secretase inhibitor L-685,458. ATF6 processing also occurred in the presence of functionally inactive dominant negative mutants of PS1 (PS1 D385N) and PS2 (PS2 D366A) that do not support endoproteolysis of beta APP and Notch. Our results therefore demonstrate that ATF6 is not a substrate for PS mediated gamma-secretase-like endoproteolysis. This finding indicates that gamma-secretase inhibitors, which are currently developed as therapeutic agents to lower the A beta burden in brains of AD patients, do not interfere with the UPR response.


In vivo detection of hyperoxia-induced pulmonary endothelial cell death using (99m)Tc-duramycin.
(99m)Tc-duramycin, DU, is a SPECT biomarker of tissue injury identifying cell death. The objective of this study is to investigate the potential of DU imaging to quantify capillary endothelial cell death in rat lung injury resulting from hyperoxia exposure as a model of acute lung injury. Rats were exposed to room air (normoxic) or >98% O2 for 48 or 60 hours. DU was injected i.v. in anesthetized rats, scintigraphy images were acquired at steady-state, and lung DU uptake was quantified from the images. Post-mortem, the lungs were removed for histological studies. Sequential lung sections were immunostained for caspase activation and endothelial and epithelial cells. Lung DU uptake increased significantly (p<0.001) by 39% and 146% in 48-hr and 60-hr exposed rats, respectively, compared to normoxic rats. There was strong correlation (r(2)=0.82, p=0.005) between lung DU uptake and the number of cleaved caspase 3 (CC3) positive cells, and endothelial cells accounted for more than 50% of CC3 positive cells in the hyperoxic lungs. Histology revealed preserved lung morphology through 48 hours. By 60 hours there was evidence of edema, and modest neutrophilic infiltrate. Rat lung DU uptake in vivo increased after just 48 hours of >98% O2 exposure, prior to the onset of any substantial evidence of lung injury. These results suggest that apoptotic endothelial cells are the primary contributors to the enhanced DU lung uptake, and support the utility of DU imaging for detecting early endothelial cell death in vivo.


A computational approach to identify point mutations associated with occult hepatitis B: significant mutations affect coding regions but not regulative elements of HBV.
Occult Hepatitis B Infection (OBI) is characterized by absence of serum HBsAg and persistence of HBV-DNA in liver tissue, with low to undetectable serum HBV-DNA. The mechanisms underlying OBI remain to be clarified. To evaluate if specific point mutations of HBV genome may be associated with OBI, we applied an approach based on bioinformatics analysis of complete genome HBV sequences. In addition, the feasibility of bioinformatics prediction models to classify HBV infections into OBI and non-OBI by molecular data was evaluated. 41 OBI and 162 non-OBI complete genome sequences were retrieved from GenBank, aligned and subjected to univariable analysis including statistical evaluation. Their S coding region was analyzed for Stop codon mutations too, while S amino acid variability could be evaluated for genotype D only, due to the too small number of available complete genome OBI sequences from other genotypes.Prediction models were derived by multivariable analysis using Logistic Regression, Rule Induction and Random Forest approaches, with extra-sample error estimation by Multiple ten-fold Cross-Validation (MCV). Models were compared by t-test on the Area Under the Receiver Operating Characteristic curve (AUC) distributions obtained from the MCV runs for each model against the best-performing model. Variations in seven nucleotide positions were significantly associated with OBI, and occurred in 11 out of 41 OBI sequences (26.8%): likely, other mutations did not reach statistical significance due to the small size of OBI dataset. All variations affected at least one HBV coding region, but none of them mapped to regulative elements. All viral proteins, with the only exception of the X, were affected. Stop codons in the S, that might account for absence of serum HBsAg, were not significantly enriched in OBI sequences. In genotype D, amino acid variability in the S was higher in OBI than non-OBI, particularly in the immunodominant region. A Random Forest prediction model showed the best performance, but all models were not satisfactory in terms of specificity, due to the small sample size of OBIs; however results are promising in the perspective of a broader dataset of complete genome OBI sequences. Data suggest that point mutations rarely occur in regulative elements of HBV, if ever, and contribute to OBI by affecting different viral proteins, suggesting heterogeneous mechanisms may be responsible for OBI, including, at least in genotype D, an escape mutation mechanism due to imperfect immune control. It appears possible to derive prediction models based on molecular data when a larger set of complete genome OBI sequences will become available.


Meningococcal meningitis: unprecedented incidence of serogroup X-related cases in 2006 in Niger.
In Niger, epidemic meningococcal meningitis is primarily caused by Neisseria meningitidis (Nm) serogroup A. However, since 2002, Nm serogroup W135 has been considered to be a major threat that has not yet been realized, and an unprecedented incidence of Nm serogroup X (NmX) meningitis was observed in 2006. Meningitis surveillance in Niger is performed on the basis of reporting of clinically suspected cases. Cerebrospinal fluid specimens are sent to the reference laboratory in Niamey, Niger. Culture, latex agglutination, and polymerase chain reaction are used whenever appropriate. Since 2004, after the addition of a polymerase chain reaction-based nonculture assay that was developed to genogroup isolates of NmX, polymerase chain reaction testing allows for the identification of Nm serogroup A, Nm serogroup B, Nm serogroup C, NmX, Nm serogroup Y, and Nm serogroup W135. From January to June 2006, a total of 4185 cases of meningitis were reported, and 2905 cerebrospinal fluid specimens were laboratory tested. NmX meningitis represented 51% of 1139 confirmed cases of meningococcal meningitis, but in southwestern Niger, it represented 90%. In the agglomeration of Niamey, the reported cumulative incidence of meningitis was 73 cases per 100,000 population and the cumulative incidence of confirmed NmX meningitis was 27.5 cases per 100,000 population (74.6 cases per 100,000 population in children aged 5-9 years). NmX isolates had the same phenotype (X : NT : P1.5), and all belonged to the same sequence type (ST-181) as the NmX isolates that were circulating in Niamey in the 1990s. Nm serogroup W135 represented only 2.1% of identified meningococci. This is, to our knowledge, the first report of such a high incidence of NmX meningitis, although an unusually high incidence of NmX meningitis was also observed in the 1990s in Niamey. The increasing incidence of NmX meningitis is worrisome, because no vaccine has been developed against this serogroup. Countries in the African meningitis belt must prepare to face this potential new challenge.


Functional domains of the floral regulator AGAMOUS: characterization of the DNA binding domain and analysis of dominant negative mutations.
The Arabidopsis MADS box gene AGAMOUS (AG) controls reproductive organ identity and floral meristem determinacy. The AG protein binds in vitro to DNA sequences similar to the targets of known MADS domain transcription factors. Whereas most plant MADS domain proteins begin with the MADS domain, AG and its orthologs contain a region N-terminal to the MADS domain. All plant MADS domain proteins share another region with moderate sequence similarity called the K domain. Neither the region (I region) that lies between the MADS and K domains nor the C-terminal region is conserved. We show here that the AG MADS domain and the I region are necessary and sufficient for DNA binding in vitro and that AG binds to DNA as a dimer. To investigate the in vivo function of the regions of AG not required for in vitro DNA binding, we introduced several AG constructs into wild-type plants and characterized their floral phenotypes. We show that transgenic Arabidopsis plants with a 35S-AG construct encoding an AG protein lacking the N-terminal region produced apetala 2 (ap2)-like flowers similar to those ectopically expressing AG proteins retaining the N-terminal region. This result suggests that the N-terminal region is not required to produce the ap2-like phenotype. In addition, transformants with a 35S-AG construct encoding an AG protein lacking the C-terminal region produced ag-like flowers, indicating that this truncated AG protein inhibits normal AG function. Finally, transformants with a 35S-AG construct encoding an AG protein lacking both K and C regions produced flowers with more stamens and carpels. The phenotypes of the AG transformants demonstrate that both the K domain and the C-terminal region have important and distinct in vivo functions. We discuss possible mechanisms through which AG may regulate downstream genes.


The jmjN and jmjC domains of the yeast zinc finger protein Gis1 interact with 19 proteins involved in transcription, sumoylation and DNA repair.
The jumonji domain is a highly conserved bipartite domain made up of two subdomains, jmjN and jmjC, which is found in many eukaryotic transcription factors. The jmjC domain was recently shown to possess the histone demethylase activity. Here we show that the jmjN and jmjC domains of the yeast zinc finger protein Gis1 interact in a two-hybrid system with 19 yeast proteins that include the RecQ helicase Sgs1, the silencing factors Esc1 and Sir4, the URI-type prefoldin Bud27 and the PIAS type SUMO ligase Nfi1/Siz2. Extensive interaction cross dependencies further suggest that the proteins form a larger complex. Consistent with this, 16 of the proteins also interact with a Bud27 two-hybrid bait, and three of them co-precipitate with TAP-tagged Gis1. The Gis1 jumonji domain can repress transcription when recruited to a promoter as a lexA fusion. This effect is dependent on both the jmjN and jmjC subdomains, as were all 19 two-hybrid interactions, indicating that the two subdomains form a single functional unit. The human Sgs1 homolog WRN also interacts with the Gis1 jumonji domain. Finally, we note that several jumonji domain interactors are related to proteins that are found in mammalian PML nuclear bodies.


Correlates of HIV Testing Among Transgender Women in Ho Chi Minh, Vietnam.
HIV prevalence among transgender women (TW) in Ho Chi Minh City is estimated at 18 %. However, no evidence-based programs or surveillance data exist in Vietnam specific to HIV testing uptake. We examined prevalence and correlates of past-year HIV testing among TW (n = 204) recruited in 2015 via snowball sampling. 59.3 % reported HIV testing in the previous year. In adjusted models, factors positively associated with HIV testing included consistent condom use during sex work with male clients; STI testing in past year; sex with casual partners in the past month; and experiences of police harassment. Factors negatively associated with recent HIV testing included daily/weekly alcohol use and post-traumatic stress symptoms. This study found significant associations between greater safety in sexual behaviors and higher rates of HIV testing. Targeted and specific services are needed for TW in Vietnam in order to address sexual risk behaviors and provide appropriate access to regular HIV testing.


Association of Solid Fuel Use With Risk of Cardiovascular and All-Cause Mortality in Rural China.
When combusted indoors, solid fuels generate a large amount of pollutants such as fine particulate matter. To assess the associations of solid fuel use for cooking and heating with cardiovascular and all-cause mortality. This nationwide prospective cohort study recruited participants from 5 rural areas across China between June 2004 and July 2008; mortality follow-up was until January 1, 2014. A total of 271 217 adults without a self-reported history of physician-diagnosed cardiovascular disease at baseline were included, with a random subset (n = 10 892) participating in a resurvey after a mean interval of 2.7 years. Self-reported primary cooking and heating fuels (solid: coal, wood, or charcoal; clean: gas, electricity, or central heating), switching of fuel type before baseline, and use of ventilated cookstoves. Death from cardiovascular and all causes, collected through established death registries. Among the 271 217 participants, the mean (SD) age was 51.0 (10.2) years, and 59% (n = 158 914) were women. A total of 66% (n = 179 952) of the participants reported regular cooking (at least weekly) and 60% (n = 163 882) reported winter heating, of whom 84% (n = 150 992) and 90% (n = 147 272) used solid fuels, respectively. There were 15 468 deaths, including 5519 from cardiovascular causes, documented during a mean (SD) of 7.2 (1.4) years of follow-up. Use of solid fuels for cooking was associated with greater risk of cardiovascular mortality (absolute rate difference [ARD] per 100 000 person-years, 135 [95% CI, 77-193]; hazard ratio [HR], 1.20 [95% CI, 1.02-1.41]) and all-cause mortality (ARD, 338 [95% CI, 249-427]; HR, 1.11 [95% CI, 1.03-1.20]). Use of solid fuels for heating was also associated with greater risk of cardiovascular mortality (ARD, 175 [95% CI, 118-231]; HR, 1.29 [95% CI, 1.06-1.55]) and all-cause mortality (ARD, 392 [95% CI, 297-487]; HR, 1.14 [95% CI, 1.03-1.26]). Compared with persistent solid fuel users, participants who reported having previously switched from solid to clean fuels for cooking had a lower risk of cardiovascular mortality (ARD, 138 [95% CI, 71-205]; HR, 0.83 [95% CI, 0.69-0.99]) and all-cause mortality (ARD, 407 [95% CI, 317-497]; HR, 0.87 [95% CI, 0.79-0.95]), while for heating, the ARDs were 193 (95% CI, 128-258) and 492 (95% CI, 383-601), and the HRs were 0.57 (95% CI, 0.42-0.77) and 0.67 (95% CI, 0.57-0.79), respectively. Among solid fuel users, use of ventilated cookstoves was also associated with lower risk of cardiovascular mortality (ARD, 33 [95% CI, -9 to 75]; HR, 0.89 [95% CI, 0.80-0.99]) and all-cause mortality (ARD, 87 [95% CI, 20-153]; HR, 0.91 [95% CI, 0.85-0.96]). In rural China, solid fuel use for cooking and heating was associated with higher risks of cardiovascular and all-cause mortality. These risks may be lower among those who had previously switched to clean fuels and those who used ventilation.


Water, ecology and health: ecosystems as settings for promoting health and sustainability.
Despite the proposed ecological and systems-based perspectives of the settings-based approach to health promotion, most initiatives have tended to overlook the fundamental nature of ecosystems. This paper responds to this oversight by proposing an explicit re-integration of ecosystems within the healthy settings approach. We make this case by focusing on water as an integrating unit of analysis. Water, on which all life depends, is not only an integral consideration for the existing healthy settings (schools, hospitals, workplaces) but also highlights the ecosystem context of health and sustainability. A focus on catchments (also know as watersheds and river basins) exemplifies the scaled and upstream/downstream nature of ecosystems and draws into sharp focus the cross-sectoral and transdisciplinary context of the social and environmental determinants of health. We position this work in relation to the converging agendas of health promotion and ecosystem management at the local, regional and global scales--and draw on evidence from international initiatives as diverse as the WHO Commission on Social Determinants of Health, and the Millennium Ecosystem Assessment. Using water as a vehicle for understanding the systemic context for human wellbeing, health promotion and disease prevention draws inevitable attention to key challenges of scale, intersectoral governance and the complementary themes of promoting resilience and preventing vulnerability. We conclude by highlighting the importance of building individual and institutional capacity for this kind of integration--equipping a new generation of researchers, practitioners and decision-makers to be conversant with the language of ecosystems, capable of systemic thought and focused on settings that can promote both health and sustainability.


Homocysteine and reactive oxygen species in metabolic syndrome, type 2 diabetes mellitus, and atheroscleropathy: the pleiotropic effects of folate supplementation.
Homocysteine has emerged as a novel independent marker of risk for the development of cardiovascular disease over the past three decades. Additionally, there is a graded mortality risk associated with an elevated fasting plasma total homocysteine (tHcy). Metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM) are now considered to be a strong coronary heart disease (CHD) risk enhancer and a CHD risk equivalent respectively. Hyperhomocysteinemia (HHcy) in patients with MS and T2DM would be expected to share a similar prevalence to the general population of five to seven percent and of even greater importance is: Declining glomerular filtration and overt diabetic nephropathy is a major determinant of tHcy elevation in MS and T2DM. There are multiple metabolic toxicities resulting in an excess of reactive oxygen species associated with MS, T2DM, and the accelerated atherosclerosis (atheroscleropathy). HHcy is associated with an increased risk of cardiovascular disease, and its individual role and how it interacts with the other multiple toxicities are presented.The water-soluble B vitamins (especially folate and cobalamin-vitamin B12) have been shown to lower HHcy. The absence of the cystathionine beta synthase enzyme in human vascular cells contributes to the importance of a dual role of folic acid in lowering tHcy through remethylation, as well as, its action of being an electron and hydrogen donor to the essential cofactor tetrahydrobiopterin. This folate shuttle facilitates the important recoupling of the uncoupled endothelial nitric oxide synthase enzyme reaction and may restore the synthesis of the omnipotent endothelial nitric oxide to the vasculature.


The internal social sustainability of sanitation infrastructure.
While the construction of sanitation infrastructure is one of humankind's greatest public health and environmental engineering achievements, its benefits are not yet enjoyed by all. In addition to the billions of people not yet reached by sanitation infrastructure, at least half of systems constructed in developing contexts are abandoned in the years following initial construction. In this research, we target the problem of postconstruction onsite sanitation infrastructure abandonment in rural Guatemala using legitimacy and status theory. Legitimacy and status are established theoretical concepts from organizational theory that reflect cultural alignment and normative support. Crisp set Qualitative Comparative Analysis (csQCA), which uses Boolean algebra to discover combinations of theoretical conditions that produce an outcome of interest, allowed us to describe the various pathways that have caused socially sustainable uptake. We find that three combinations of legitimacy and status theory explain 85% of household cases at a consistency of 0.97. The most practically useful pathway covers 50% of household cases and shows that the combination of consequential legitimacy (a moral understanding of outcomes) and comprehensibility legitimacy (a cognitive model connecting outcomes to processes) is a powerful way to achieve socially sustainable sanitation infrastructure.


Particulate matter emissions of four types of one cigarette brand with and without additives: a laser spectrometric particulate matter analysis of secondhand smoke.
Inhaled particulate matter (PM) in secondhand smoke (SHS) is deleterious for smokers and non-smokers. Different additives in cigarettes might effect the amount of PM. This study aimed to assess the influence of additives on the PM emissions from different cigarette types in SHS. An experimental study of PM measuring in SHS of cigarettes without exposition of any person. The concentrations of PM (PM10, PM2.5 and PM1) in SHS of four different types of cigarettes of the brand Lucky Strike, two types with additives (Original Red, Original Blue) and two types without additives (Straight Red, Straight Blue), in comparison to the reference cigarette 3R4F were analysed. An automatic environmental tobacco smoke emitter generated SHS in an enclosed space with a volume of 2.88 m3. PM was measured with a laser aerosol spectrometer (Grimm model 1.109). Afterwards, the measuring values of the four Lucky Strike brands and the reference cigarette were statistically evaluated and visualised. Lucky Strike Straight Blue, a cigarette type without additives and lower tar amount, showed 10% to 25% lower PM mean values compared with the other tested Lucky Strike products, but 21% (PM1) respectively 27% (PM2.5,PM10) higher mean values than the reference cigarette. The PM mean of all measured smoke-free baseline values (clean air) was 1.6 µg/m³. It increased up to about 1800 µg/m³ for the reference cigarette and up to about 3070 µg/m³ for the Lucky Strike Original Blue. The findings of this study show the massive increase of PM amount by smoking cigarettes in enclosed spaces and suggest that additives in tobacco products increase the PM amount in SHS. For validation, further comparative studies are necessary focusing on the comparison of the PM concentration of cigarettes with and without additives. Due to the exposure to SHS, 890 000 people die each year worldwide. PM in SHS endangers the health of both non-smokers and smokers. This study considers the effect of additives like aromatics and humectant agents in cigarettes on PM in SHS. Do additives in tobacco products increase the amount of PM?


Simulating 3-D lung dynamics using a programmable graphics processing unit.
Medical simulations of lung dynamics promise to be effective tools for teaching and training clinical and surgical procedures related to lungs. Their effectiveness may be greatly enhanced when visualized in an augmented reality (AR) environment. However, the computational requirements of AR environments limit the availability of the central processing unit (CPU) for the lung dynamics simulation for different breathing conditions. In this paper, we present a method for computing lung deformations in real time by taking advantage of the programmable graphics processing unit (GPU). This will save the CPU time for other AR-associated tasks such as tracking, communication, and interaction management. An approach for the simulations of the three-dimensional (3-D) lung dynamics using Green's formulation in the case of upright position is taken into consideration. We extend this approach to other orientations as well as the subsequent changes in breathing. Specifically, the proposed extension presents a computational optimization and its implementation in a GPU. Results show that the computational requirements for simulating the deformation of a 3-D lung model are significantly reduced for point-based rendering.


Rare cytogenetic abnormalities in myelodysplastic syndromes.
The karyotype represents one of the main cornerstones for the International Prognostic Scoring System (IPSS) and the revised IPSS-R (IPSS-R) that are most widely used for prognostication in patients with myelodysplastic syndromes (MDS). The most frequent cytogenetic abnormalities in MDS, i.e. del(5q), -7/del(7q), +8, complex karyotypes, or -Y have been extensively explored for their prognostic impact. The IPSS-R also considers some less frequent abnormalities such as del(11q), isochromosome 17, +19, or 3q abnormalities. However, more than 600 different cytogenetic categories had been identified in a previous MDS study. This review aims to focus interest on selected rare cytogenetic abnormalities in patients with MDS. Examples are numerical gains of the chromosomes 11 (indicating rapid progression), of chromosome 14 or 14q (prognostically intermediate to favorable), -X (in females, with an intermediate prognosis), or numerical abnormalities of chromosome 21. Structural abnormalities are also considered, e.g. del(13q) that is associated with bone marrow failure syndromes and favorable response to immunosuppressive therapy. These and other rare cytogenetic abnormalities should be integrated into existing prognostication systems such as the IPSS-R. However, due to the very low number of cases, this is clearly dependent on international collaboration. Hopefully, this article will help to inaugurate this process.


[Diagnostic references in osteo-archeology. The contribution of historical medical data].
Some examples of anatomo-radio-clinical (or medico-historical) collation, studied in the Dupuytren Museum of Paris, among osteoplastic conditions, including hyperostosis frontalis interna, fibrous dysplasia of bone, osteochondromas, angioma of bone and osteosarcomas, demonstrate how that triad allows to constitute more secure guidelines for the archaeologist who have not clinical data at his disposal. So, they encourage to support the defence and maintenance actions of such anatomico-pathological museums.


Absence of a correlation of androgen receptor gene CAG repeat length and prostate cancer risk in an African-American population.
Shorter androgen receptor gene CAG repeat length has been associated with an increased risk of prostate cancer, an earlier age of onset, and more advanced stage of disease. Studies comparing the distribution of CAG repeat lengths within different populations have reported that racial groups with higher prostate cancer incidence also have shorter CAG repeat lengths. We evaluated CAG repeat length in 685 black men in Louisiana, South Carolina, and the District of Columbia who were participating in prostate cancer screening, comparing the 118 who were diagnosed with prostate cancer with 567 who had normal serum prostate-specific antigen levels and no evidence of cancer on digital rectal examination. The median CAG repeat length was 21 among cases and 19 among controls (P = 0.11). Cases were significantly older than controls, with a median age of 68 years compared with 54 years (P < 0.0001). After adjusting for age, we found no association between prostate cancer risk and CAG repeat length (odds ratio, 1.05; 95% CI, 0.98-1.13; P = 0.16). Dividing CAG repeat lengths into septiles and calculating the odds ratio for each revealed no specific repeat-length range with a significantly elevated or depressed risk of prostate cancer, but a trend test showed a significant association between longer CAG repeat lengths and an elevated risk of prostate cancer (P = 0.02). Neither grade nor stage was associated with CAG repeat length. This study confirms earlier reports that black men have shorter CAG repeat lengths than reported white and Asian populations. We did not find an increased risk of prostate cancer among black men with fewer CAG repeats.


Staphylococcus aureus in a neonatal care center: methicillin-susceptible strains should be a main concern.
In the context of a methicillin-susceptible Staphylococcus aureus (MSSA) outbreak, we aimed to improve our knowledge of S. aureus (SA) epidemiology in the neonatal care center (NCC) of a tertiary care teaching hospital. We performed a complete one-year review of SA carrier, colonized or infected patients. Monthly prevalence and incidence of SA intestinal carriage, colonization and infection were calculated and the types of infection analysed. During the MSSA outbreak, strains were studied for antimicrobial resistance, content of virulence genes and comparative fingerprint in Pulsed-Field Gel Electrophoresis. Hand hygiene and catheter-related practices were assessed by direct observational audits. Environmental investigation was performed in search of a SA reservoir. Epidemiological analyses showed 2 or 3 prevalence peaks on a background of SA endemicity. In the NCC, during 2009, overall MSSA prevalence did not decrease below 5.5%, while mean MRSA prevalence was about 1.53%. Analysis of infection cases revealed that the outbreak corresponded to the emergence of catheter-related infections and was probably related to the relaxation in infection control practices in a context of high colonization pressure. Health care workers' white coats appeared as a potential environmental reservoir that could perpetuate SA circulation in the ward. This report emphasizes the importance of integrating MSSA along with methicillin-resistant SA in a program of epidemiological surveillance in the NCC.


Prescribers and pharmaceutical representatives: why are we still meeting?
Research suggests that pharmaceutical marketing influences prescribing and may cause cognitive dissonance for prescribers. This work has primarily been with physicians and physician-trainees. Questions remain regarding why prescribers continue to meet with pharmaceutical representatives (PRs). To describe the reasons that prescribers from various health professions continue to interact with PRs despite growing evidence of the influence of these interactions. Multi-disciplinary focus groups with 61 participants held in practice settings and at society meetings. Most prescribers participating in our focus groups believe that overall PR interactions are beneficial to patient care and practice health. They either trust the information from PRs or feel that they are equipped to evaluate it independently. Despite acknowledgement of study findings to the contrary, prescribers state that they are able to effectively manage PR interactions such that their own prescribing is not adversely impacted. Prescribers describe few specific strategies or policies for these interactions, and report that policies are not consistently implemented with all members of a clinic or institution. Some prescribers perceive an inherent contradiction between academic centers and national societies receiving money from pharmaceutical companies, and then recommending restriction at the level of the individual prescriber. Prescribers with different training backgrounds present a few novel reasons for these meetings. Despite evidence that PR detailing influences prescribing, providers from several health professions continue to believe that PR interactions improve patient care, and that they can adequately evaluate and filter information presented to them by PRs. Focus group comments suggest that cultural change is necessary to break the norms that exist in many settings. Applying policies consistently, considering non-physician members of the healthcare team, working with trainees, restructuring the current primary care model and offering convenient, individualized, non-biased educational options may aid success.


The nucleoporin RanBP2 has SUMO1 E3 ligase activity.
Posttranslational modification with SUMO1 regulates protein/protein interactions, localization, and stability. SUMOylation requires the E1 enzyme Aos1/Uba2 and the E2 enzyme Ubc9. A family of E3-like factors, PIAS proteins, was discovered recently. Here we show that the nucleoporin RanBP2/Nup358 also has SUMO1 E3-like activity. RanBP2 directly interacts with the E2 enzyme Ubc9 and strongly enhances SUMO1-transfer from Ubc9 to the SUMO1 target Sp100. The E3-like activity is contained within a 33 kDa domain of RanBP2 that lacks RING finger motifs and does not resemble PIAS family proteins. Our findings place SUMOylation at the cytoplasmic filaments of the NPC and suggest that, at least for some substrates, modification and nuclear import are linked events.


Defining the genes for the final steps in biosynthesis of the complex polyketide antibiotic mupirocin by Pseudomonas fluorescens NCIMB10586.
The mupirocin trans-AT polyketide synthase pathway, provides a model system for manipulation of antibiotic biosynthesis. Its final phase involves removal of the tertiary hydroxyl group from pseudomonic acid B, PA-B, producing the fully active PA-A in a complex series of steps. To further clarify requirements for this conversion, we fed extracts containing PA-B to mutants of the producer strain singly deficient in each mup gene. This additionally identified mupM and mupN as required plus the sequence but not enzymic activity of mupL and ruled out need for other mup genes. A plasmid expressing mupLMNOPVCFU + macpE together with a derivative of the producer P. fluorescens strain NCIMB10586 lacking the mup cluster allowed conversion of PA-B to PA-A. MupN converts apo-mAcpE to holo-form while MupM is a mupirocin-resistant isoleucyl tRNA synthase, preventing self-poisoning. Surprisingly, the expression plasmid failed to allow the closely related P. fluorescens strain SBW25 to convert PA-B to PA-A.


Vortex configuration flow cell based on low-temperature cofired ceramics as a compact chemiluminescence microsystem.
The integration of optical detection methods in continuous flow microsystems can highly extend their range of application, as long as some negative effects derived from their scaling down can be minimized. Downsizing affects to a greater extent the sensitivity of systems based on absorbance measurements than the sensitivity of those based on emission ones. However, a careful design of the instrumental setup is needed to maintain the analytical features in both cases. In this work, we present the construction and evaluation of a simple miniaturized optical system, which integrates a novel flow cell configuration to carry out chemiluminescence (CL) measurements using a simple photodiode. It consists of a micromixer based on a vortex structure, which has been constructed by means of the low-temperature cofired ceramics (LTCC) technology. This mixer not only efficiently promotes the CL reaction due to the generated high turbulence but also allows the detection to be carried out in the same area, avoiding intensity signal losses. As a demonstration, a flow injection system has been designed and optimized for the detection of cobalt(II) in water samples. It shows a linear response between 2 and 20 microM with a correlation of r > 0.993, a limit of detection of 1.1 microM, a repeatability of RSD = 12.4%, and an analysis time of 17 s. These results demonstrate the suitability of the proposal to the determination of compounds involved in CL reactions by means of an easily constructed versatile device based on low-cost instrumentation.


Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens.
The rapid emergence of SARS-CoV-2, the causative agent of COVID-19, and its dissemination globally has caused an unprecedented strain on public health. Animal models are urgently being developed for SARS-CoV-2 to aid rational design of vaccines and therapeutics. Immunohistochemistry and in situ hybridisation techniques that facilitate reliable and reproducible detection of SARS-CoV and SARS-CoV-2 viral products in formalin-fixed paraffin-embedded (FFPE) specimens would be of great utility. A selection of commercial antibodies generated against SARS-CoV spike protein and nucleoprotein, double stranded RNA, and RNA probe for spike genes were evaluated for the ability to detect FFPE infected cells. We also tested both heat- and enzymatic-mediated virus antigen retrieval methods to determine the optimal virus antigen recovery as well as identifying alternative retrieval methods to enable flexibility of IHC methods. In addition to using native virus infected cells as positive control material, the evaluation of non-infected cells expressing coronavirus (SARS, MERS) spike as a biosecure alternative to assays involving live virus was undertaken. Optimized protocols were successfully applied to experimental animal-derived tissues. The diverse techniques for virus detection and control material generation demonstrated in this study can be applied to investigations of coronavirus pathogenesis and therapeutic research in animal models.


Functional status of patients with metastatic spinal cord compression.
This prospective study was conducted to evaluate the functional status and clinical features of inpatients with metastatic spinal cord compression and to explore possible associated factors and the correlation with neurological deficits. Cases were identified through an active search in the hospital units of the National Cancer Institute. For 47 patients, clinical and demographic characteristics were collected; functional status was measured by the Functional Independence Measure; modified Tokuhashi score was used to predict survival time; neurological deficits were measured using the American Spinal Injury Association Impairment Scale; and general condition was evaluated using the Karnofsky Performance Status Scale. The correlation between the level of neurological deficits and functional status was calculated using the Spearman correlation test. The difference in the functional independence score according to neurological status was assessed with the Kruskall-Wallis test. The degree of functional status was associated with Karnofsky Performance Status, level of neurological deficits, ability to walk, survival time, and skin color (p < 0.05). There was difference in the functional status (p = 0.004) and in the motor domain (p = 0.001) according to the level of neurological deficits. The correlation of the Functional Independence Measure with level of neurological deficits and with Karnofsky Performance Status was moderate (r = 0.46, p < 0.02). The Functional Independence Measure was adequate for evaluating the functional status of patients with metastatic spinal cord compression. Palliative rehabilitation should be integrated early and strategies should be linked to the prognosis of survival.


Structural characterization of SLYM - a 4 th meningeal membrane.
Traditionally, the meninges are described as 3 distinct layers, dura, arachnoid and pia. Yet, the classification of the connective meningeal membranes surrounding the brain is based on postmortem macroscopic examination. Ultrastructural and single cell transcriptome analyses have documented that the 3 meningeal layers can be subdivided into several distinct layers based on cellular characteristics. We here re-examined the existence of a 4 th meningeal membrane, S ubarachnoid Ly mphatic-like M embrane or SLYM in Prox1-eGFP reporter mice. Imaging of freshly resected whole brains showed that SLYM covers the entire brain and brain stem and forms a roof shielding the subarachnoid cerebrospinal fluid (CSF)-filled cisterns and the pia-adjacent vasculature. Thus, SLYM is strategically positioned to facilitate periarterial influx of freshly produced CSF and thereby support unidirectional glymphatic CSF transport. Histological analysis showed that, in spinal cord and parts of dorsal cortex, SLYM fused with the arachnoid barrier layer, while in the basal brain stem typically formed a 1-3 cell layered membrane subdividing the subarachnoid space into two compartments. However, great care should be taken when interpreting the organization of the delicate leptomeningeal membranes in tissue sections. We show that hyperosmotic fixatives dehydrate the tissue with the risk of shrinkage and dislocation of these fragile membranes in postmortem preparations.


European Registry on Helicobacter pylori management (Hp-EuReg): patterns and trends in first-line empirical eradication prescription and outcomes of 5 years and 21 533 patients.
The best approach for Helicobacter pylori management remains unclear. An audit process is essential to ensure clinical practice is aligned with best standards of care. International multicentre prospective non-interventional registry starting in 2013 aimed to evaluate the decisions and outcomes in H. pylori management by European gastroenterologists. Patients were registered in an e-CRF by AEG-REDCap. Variables included demographics, previous eradication attempts, prescribed treatment, adverse events and outcomes. Data monitoring was performed to ensure data quality. Time-trend and geographical analyses were performed. 30 394 patients from 27 European countries were evaluated and 21 533 (78%) first-line empirical H. pylori treatments were included for analysis. Pretreatment resistance rates were 23% to clarithromycin, 32% to metronidazole and 13% to both. Triple therapy with amoxicillin and clarithromycin was most commonly prescribed (39%), achieving 81.5% modified intention-to-treat eradication rate. Over 90% eradication was obtained only with 10-day bismuth quadruple or 14-day concomitant treatments. Longer treatment duration, higher acid inhibition and compliance were associated with higher eradication rates. Time-trend analysis showed a region-dependent shift in prescriptions including abandoning triple therapies, using higher acid-inhibition and longer treatments, which was associated with an overall effectiveness increase (84%-90%). Management of H. pylori infection by European gastroenterologists is heterogeneous, suboptimal and discrepant with current recommendations. Only quadruple therapies lasting at least 10 days are able to achieve over 90% eradication rates. European recommendations are being slowly and heterogeneously incorporated into routine clinical practice, which was associated with a corresponding increase in effectiveness.


Mortality of farmers and farmers' wives in England and Wales 1979-80, 1982-90.
To examine the mortality patterns of male and female farmers and farmers' wives in England and Wales. Information on all deaths in England and Wales at ages 20-74 during the periods 1979-80 and 1982-90 was obtained from the Office of Population Censuses and Surveys. Proportional mortality ratios (PMRs) and proportional cancer mortality ratios (PCMRs) were used to compare the mortality of farmers with that of the general working population, and of farmers' wives with wives of all working men. Farmers and farmers' wives had high mortality from accidents and suicide and from certain respiratory diseases. Mortality from hernia was also raised. Deaths from cancer were generally below expectations, but the PMR for prostatic cancer was 112 (95%CI 106-118). The PMRs and PCMRs for oesophageal cancer were significantly increased in male farmers from two counties where cider is produced. The occupational hazards of farming continue to be associated with excess mortality, and most of the risks extend also to farmers' wives. Action is needed to reduce deaths, particularly from accidents and suicide.


Genetic and mass spectrometric tools for elucidating the physiological function(s) of cytochrome P450 enzymes from Mycobacterium tuberculosis.
Tuberculosis remains a leading cause of human mortality. The emergence of strains of Mycobacterium tuberculosis (Mtb), the causative agent, that are resistant to first- and second-line antitubercular drugs urges the development of new therapeutics. The genome of Mtb encodes 20 cytochrome P450 enzymes, at least some of which are potential candidates (CYP121, CYP125, and CYP128) for drug targeting. In this regard, we examined the specific role of CYP125 in the cholesterol degradation pathway, using genetic and mass spectrometric approaches. The analysis of lipid profiles from Mtb cells grown on cholesterol revealed that CYP125, by virtue of its C26-monooxygenase activity, is essential for cholesterol degradation, and, consequently, for the incorporation of side-chain carbon atoms into cellular lipids, as evidenced by an increase in the mass of the methyl-branched phthiocerol dimycocerosates (PDIM). Moreover, this work also led to the identification of cholest-4-en-3-one as a source of cellular toxicity. Herein, we describe the experimental procedures that led to elucidation of the physiological function of CYP125. A similar approach can be used to study other important Mtb P450 enzymes.


Increased development of T-bet+CD11c+ B cells predisposes to lupus in females: Analysis in BXD2 mouse and genetic crosses.
Cardinal features of lupus include elevated B cell activation and autoantibody production with a female sex preponderance. We quantified interactions of sex and genetic variation on the development of autoimmune B-cell phenotypes and autoantibodies in the BXD2 murine model of lupus using a cohort of backcrossed progeny (BXD2 x C57BL/6J) x BXD2. Sex was the key factor leading to increased total IgG, IgG2b, and autoantibodies. The percentage of T-bet+CD11c+ IgD+ activated naive B cells (aNAV) was higher in females and was associated with increased T-bet+CD11c+ IgD- age-related B cells, Fas+GL7+ germinal center B cells, Cxcr5-Icos+ peripheral T-helper cells, and Cxcr5+Icos+ follicular T-helper cells. IFN-β was elevated in females. Variation in aNAV cells was mapped to Chr 7 in a locus that shows significant interactions between the female sex and heterozygous B/D variant. Our results suggest that activation of naive B cells forms the basis for the female-predominant development of autoantibodies in lupus-susceptible BXD2 mice.


Risk of Blindness Among Patients With Diabetes and Newly Diagnosed Diabetic Retinopathy.
To evaluate the association between initial diabetic retinopathy (DR) severity/risk of blindness in patients with newly diagnosed DR/good vision in the U.S. This retrospective cohort study evaluated adult patients with good vision (20/40 or better) and newly diagnosed DR between 1 January 2013 and 31 December 2017 (index date) in the American Academy of Ophthalmology's Intelligent Research in Sight (IRIS) Registry. The primary exposure of interest was DR severity at index: mild nonproliferative DR (NPDR), moderate NPDR, severe NPDR, and proliferative DR (PDR). The main outcome measure was development of sustained blindness (SB), defined as study eyes with Snellen visual acuity readings of 20/200 or worse at two separate visits ≥3 months apart that did not improve beyond 20/100. Among 53,535 eligible eyes (mean follow-up 662.5 days), 678 (1.3%) eyes developed SB. Eyes with PDR at index represented 10.5% (5,629 of 53,535) of the analysis population but made up 26.5% (180 of 678) of eyes that developed SB. Kaplan-Meier analysis revealed that eyes with moderate NPDR, severe NPDR, and PDR at index were 2.6, 3.6, and 4.0 times more likely, respectively, to develop SB after 2 years of DR diagnosis versus eyes with mild DR at index. In a Cox proportional hazards model adjusted for index characteristics/development of ocular conditions during follow-up, eyes with PDR had an increased risk of developing SB versus eyes with mild NPDR at index (hazard ratio 2.26 [95% CI 2.09-2.45]). In this longitudinal ophthalmologic registry population involving eyes with good vision, more advanced DR at first diagnosis was a significant risk factor for developing SB.


Phylogenetic relationships in Ephedra (Ephedraceae) inferred from chloroplast and nuclear DNA sequences.
Sequences of the nuclear ribosomal DNA internal transcribed spacer region 1 and the chloroplast-encoded genes maturase K and ribulose-1,5 biphosphate carboxylase large subunit were obtained from species of Ephedra (Ephedraceae) representing the geographic range and morphological diversity of the genus. Phylogenetic analyses of the DNA data indicate that relationships within the genus are better predicted by geographic region of origin than by ovulate cone characters. The sampled species with dry, winged (versus fleshy) ovulate cone bracts or single-seeded cones do not form monophyletic groups and therefore the previous classification systems of Ephedra based on these aspects of bract morphology appear to be largely unnatural. Three groups were identified among the Old World species studied, one comprising European and Mediterranean species and two including only Asian species. The sequence data suggest a possible early divergence of a New World clade of Ephedra from among the Old World groups. The South American species form a distinct clade apparently related to one of two groups of North American species, which accords with a frequent floristic pattern of close relationships between species groups in western South America and southwestern North America.


Self-esteem and health-risk behaviors among Turkish late adolescents.
The current study investigated (a) the relationship between self-esteem and health-risk behaviors and use of cigarettes, alcohol, and drugs, and (b) the gender differences in self-esteem and health-risk behaviors among a group of 243 late adolescents (124 males, 119 females) using a cross-sectional survey design. The age range of the participants was 17 to 24 with a mean age of 20.43 (SD = 1.21). Participants completed the Rosenberg Self-Esteem Scale and a self-report questionnaire including items about demographic characteristics and participation within a range of health-risk behaviors. The findings of the study revealed that self-esteem was negatively associated with alcohol and illicit drug use; however, these results did not suggest any significant relationship between self-esteem and smoking cigarettes. Comparisons between males and females did not indicate any gender differences on the self-esteem scale. On the other hand, significant gender differences were found on cigarette and drug use with males reporting more cigarette and drug use. Limitations of the study and possible implications for counseling practice are discussed.


Reactions of healthy persons and persons suffering from allergic rhinitis when exposed to office dust.
Reactions to airborne office dust among healthy subjects and subjects suffering from allergic rhinitis were investigated. Twelve healthy and 11 subjects suffering from allergic rhinitis were exposed to clean air [17 (SD 2) microg/m3] and office dust [439 (SD 68) microg/m3] for 245 minutes. The effect measurements included subjective sensations (questionnaire and potentiometer ratings), mood scale, peak flow, bronchial provocation with histamine using forced expiratory volume in 1 second as the effect measure, nasal mucosal swelling, tear film stability, epithelial damage, foam formation in the eye canthus, threshold for eye irritation with carbon dioxide, eye redness, cellular content of conjunctival fluid, and an addition test for distraction. As many investigations were made and as many statistical analyses (including subgroup analyses) were carried out, the risk of mass significance appeared. This problem was dealt with using the Bonferroni correction for multiple significance tests. The mean ratings of the potentiometer were higher (the subjects showed more irritation) during the dust exposure. The objective investigations showed only indications of effects of dust exposure, and some of the indications were in biologically unexplainable directions. No difference in the reactions to dust was observed between the healthy subjects and the subjects suffering from allergic rhinitis. Dust does not seem to have objective or subjective effects on humans, as only indications of dust effects were found. Subjects suffering from allergic rhinitis do not appear to be a risk group in relation to dust exposure.


Adolescent and young adult oncology, version 2.2014.
The NCCN Guidelines Insights on Adolescent and Young Adult (AYA) Oncology discuss the fertility and endocrine issues that are relevant to the management of AYA patients with cancer. Fertility preservation should be an essential part in the treatment of AYA patients with cancer. The NCCN Guidelines recommend discussion of fertility preservation and contraception before the start of treatment. Oophoropexy and embryo cryopreservation are the 2 established options for fertility preservation in women. Semen cryopreservation before the start of treatment is the most reliable and well-established method of preserving fertility in men. AYA women with cancer also have unique contraception needs, depending on the type of cancer, its treatment, and treatment-related complications. Management of cancer during pregnancy poses significant diagnostic and therapeutic challenges for both the patient and the physician. AYA women diagnosed with cancer during pregnancy require individualized treatment from a multidisciplinary team involving medical, surgical, radiation, and gynecologic oncologists; obstetricians; and perinatologists.


A pilot study to investigate the short-term effects of specific soft tissue massage on upper cervical movement impairment in patients with cervicogenic headache.
Upper cervical movement impairment and muscle dysfunction have been identified as core components of cervicogenic headache (CGH) pathogenesis. The purpose of this single-group pre-post test pilot study was to investigate the short-term effects of a specific soft tissue massage (SSTM) intervention to the cervical spine on range of upper cervical motion. Eight subjects (mean age 28.1 years) with published criteria of CGH (mean history of headache for 7.1 years) were investigated. Range of rotation of the upper cervical spine to the left and right was determined by the flexion-rotation test. Movement was assessed in three phases: pre-intervention, intervention, and post-intervention. The SSTM intervention consisted of an 8-minute soft tissue massage to the cervical muscles bilaterally. Pre-intervention measures of flexion-rotation test range of motion prior to the intervention over two assessment points were consistent. In contrast, a repeated measures analysis of variance revealed a significant improvement in range of rotation to the left and right after the first (P<0.01), second (P<0.01), but not third intervention (P = 0.19), from an average range of 27.5° at baseline to 45.9° at the third treatment session. After the 2-week post-intervention phase, range of motion remained stable without decline, and was considered full range. This pilot study provides preliminary evidence of the potential for SSTM to improve, at least in the short-term, upper cervical range of motion in people with CGH.


A written self-help intervention for depressed adults comparing behavioural activation combined with physical activity promotion with a self-help intervention based upon behavioural activation alone: study protocol for a parallel group pilot randomised controlled trial (BAcPAc).
Challenges remain to find ways to support patients with depression who have low levels of physical activity (PA) to overcome perceived barriers and enhance the perceived value of PA for preventing future relapse. There is an evidence-base for behavioural activation (BA) for depression, which focuses on supporting patients to restore activities that have been avoided, but practitioners have no specific training in promoting PA. We aimed to design and evaluate an integrated BA and PA (BAcPAc) practitioner-led, written, self-help intervention to enhance both physical and mental health. This study is informed by the Medical Research Council Complex Intervention Framework and describes a protocol for a pilot phase II randomised controlled trial (RCT) to test the feasibility and acceptability of the trial methods to inform a definitive phase III RCT. Following development of the augmented written self-help intervention (BAcPAc) incorporating behavioural activation with physical activity promotion, depressed adults are randomised to receive up to 12 sessions over a maximum of 4 months of either BAcPAc or behavioural activation alone within a written self-help format, which represents treatment as usual. The study is located within two 'Improving Access to Psychological Therapies' services in South West England, with both written self-help interventions supported by mental health paraprofessionals. Measures assessed at 4, 9, and 12 month follow-up include the following: CIS-R, PHQ-9, accelerometer recorded (4 months only) and self-reported PA, body mass index, blood pressure, Insomnia Severity Index, quality of life, and health and social care service use. Process evaluation will include analysis of recorded support sessions and patient and practitioner interviews. At the time of writing the study has recruited 60 patients. The feasibility outcomes will inform a definitive RCT to assess the clinical and cost-effectiveness of the augmented BAcPAc written self-help intervention to reduce depression and depressive relapse, and bring about improvements across a range of physical health outcomes. Current Controlled Trials ISRCTN74390532, 26.03.2013.


Self-propulsion of aluminum particle-coated Janus droplet in alkaline solution.
Janus droplet motion powered by gas bubbles is studied in this paper. The Janus droplets were fabricated by partially covering one side of an oil droplet with aluminum particles under gravity effect. By placing the Janus droplet in an alkaline solution, the reaction between the Al particles and OH- generates H2 gas, which emits from the particle-coated side of the droplet as bubbles. Hence, the droplet is propelled to move in the opposite direction. In this research, the effects of several factors, time, pH, particle coverage and surfactant, on the motion of the Janus droplet were studied. The experimental results indicate that as time progresses, the droplet motion experiences three periods: initial development stage, stable stage and decline stage. The speed of the droplet increases with the pH value and the particle coverage. Comparing with the other three surfactants, Tween 20, CTAB and SDS, the surfactant Triton X-100 is the best choice for generating the spontaneous motion of the Janus droplets. Furthermore, the directional motion of the Janus droplets was examined, and the results show that the controllable transportation of the Janus droplets can be accomplished by externally applied DC electric field. The spontaneous motion of the droplets offers great promise in chemical and biological applications.


[Study on correlation between alcohol consumption and obesity in adults in China].
Objective: To evaluate the correlation between alcohol consumption and obesity in adults in China. Methods: The information about alcohol consumption were collected at the baseline survey of the China Kadoorie Biobank. The general obesity and central obesity were defined by BMI and waist circumference (WC) respectively. Logistic regression model was employed to examine the relationship of drinking behavior with general obesity and central obesity. Results: A total of 249 873 adults were included. A J-shaped relationship was observed between alcohol consumption and obesity measurement index (BMI and WC) in men. Compared with non-drinkers, the proportion of general obesity and central obesity were lower in light drinkers (men: OR=0.65, 95%CI: 0.59-0.71 and OR=0.93, 95%CI: 0.88-0.98; women: OR=0.77, 95%CI: 0.65-0.91 and OR=0.89, 95%CI: 0.80-0.99). In men, the proportion of general obesity and central obesity was highest in heavy drinkers (OR=1.21, 95%CI: 1.12-1.32; OR=1.33, 95%CI: 1.27-1.40). BMI and WC were higher in those with a drinking frequency of 3-5 d/week, with largest of proportion of central obesity (men: OR=1.23, 95%CI: 1.16-1.31; women: OR=1.13, 95%CI: 0.99-1.28). The risk for central obesity in men who began drinking every week before 20 years old was 1.24 times higher than non-drinkers (95%CI: 1.16-1.33). Those who drank beer had lower proportion of general obesity (men: OR=0.74, 95%CI: 0.67-0.82; women: OR=0.54, 95%CI: 0.43-0.68). Conclusion: The proportion of obesity was lower in light drinkers but higher in heavy drinkers; and the earlier drinking started, the higher the risk for obesity was.


Spatiotemporal Distribution of GABAA Receptor Subunits Within Layer II of Mouse Medial Entorhinal Cortex: Implications for Grid Cell Excitability.
GABAergic parvalbumin-expressing (PV+) interneurons provide powerful inhibitory modulation of grid cells in layer II of the medial entorhinal cortex (MEC LII). However, the molecular machinery through which PV+ cells regulate grid cell activity is poorly defined. PV+ interneurons impart inhibitory modulation primarily via GABA-A receptors (GABAARs). GABAARs are pentameric ion channels assembled from a repertoire of 19 subunits. Multiple subunit combinations result in a variety of receptor subtypes mediating functionally diverse postsynaptic inhibitory currents. Whilst the broad expression patterns of GABAAR subunits within the EC have been reported, those expressed by individual MEC LII cell types, in particular grid cells candidates, stellate and pyramidal cells, are less well described. Stellate and pyramidal cells are distinguished by their selective expression of reelin (RE+) and calbindin (CB+) respectively. Thus, the overall aim of this study was to provide a high resolution analysis of the major (α and γ) GABAAR subunits expressed in proximity to somato-dendritic PV+ boutons, on RE+ and CB+ cells, using immunohistochemistry, confocal microscopy and quantitative RT-PCR (qPCR). Clusters immunoreactive for the α1 and γ2 subunits decorated the somatic membranes of both RE+ and CB+ cells and were predominantly located in apposition to clusters immunoreactive for PV and vesicular GABA transporter (VGAT), suggesting expression in GABAergic synapses innervated by PV interneurons. Although intense α2 subunit-immunopositive clusters were evident in hippocampal fields located in close proximity to the EC, no specific signal was detected in MEC LII RE+ and CB+ profiles. Immunoreactivity for the α3 subunit was detected in all RE+ somata. In contrast, only a sub-population of CB+ cells was α3 immunopositive. These included CB-α3 cells which were both PV+ and PV-. Furthermore, α3 subunit mRNA and immunofluorescence decreased significantly between P 15 and P 25, a period implicated in the functional maturation of grid cells. Finally, α5 subunit immunoreactivity was detectable only on CB+ cells, not on RE+ cells. The present data demonstrates that physiologically distinct GABAAR subtypes are selectively expressed by CB+ and RE+ cells. This suggests that PV+ interneurons could utilize distinct postsynaptic signaling mechanisms to regulate the excitability of these different, candidate grid cell sub-populations.


Antimicrobial susceptibility of Arcanobacterium pyogenes isolated from the lungs of white-tailed deer (Odocoileus virginianus) with pneumonia.
In vitro susceptibilities of 29 strains of Arcanobacterium pyogenes isolated from lung lesions of white-tailed deer (Odocoileus virginianus) with pneumonia were determined using the broth microdilution method to ascertain efficacious treatment options for pneumonic white-tailed deer. All 29 A. pyogenes strains tested were susceptible to ceftiofur, spectinomycin, tiamulin, and trimethoprim-sulfamethoxazole but were resistant to both danofloxacin and sulfadimethoxine. Likewise, all 29 isolates were either fully susceptible or intermediately susceptible to gentamicin (25 susceptible; 4 intermediate) and tulathromycin (25 susceptible; 4 intermediate). At least one isolate of A. pyogenes tested was resistant to ampicillin, chlortetracycline, clindamycin, enrofloxacin, florfenicol, oxytetracycline, penicillin, and tilmicosin suggesting their ineffectiveness in treating A. pyogenes-associated lung infections in white-tailed deer. Minimum inhibitory concentration (MIC) data for tylosin and neomycin could not be interpreted due to unavailability of Clinical and Laboratory Standards Institute (CLSI)-approved breakpoints for these 2 agents. In summary, based on MIC values, ceftiofur, spectinomycin, tiamulin, and trimethoprim-sulfamethoxazole are more efficacious than other antimicrobial agents for treating A. pyogenes-related pneumonia in white-tailed deer. However, ceftiofur may be preferred over the other 4 drugs as it is being widely used to treat respiratory disease in cattle and other animal species, as well as is available for single dose parenteral administration.


Predicting the sequence specificities of DNA- and RNA-binding proteins by deep learning.
Knowing the sequence specificities of DNA- and RNA-binding proteins is essential for developing models of the regulatory processes in biological systems and for identifying causal disease variants. Here we show that sequence specificities can be ascertained from experimental data with 'deep learning' techniques, which offer a scalable, flexible and unified computational approach for pattern discovery. Using a diverse array of experimental data and evaluation metrics, we find that deep learning outperforms other state-of-the-art methods, even when training on in vitro data and testing on in vivo data. We call this approach DeepBind and have built a stand-alone software tool that is fully automatic and handles millions of sequences per experiment. Specificities determined by DeepBind are readily visualized as a weighted ensemble of position weight matrices or as a 'mutation map' that indicates how variations affect binding within a specific sequence.


Inverse probability of censoring weighted estimates of Kendall's τ for gap time analyses.
In life history studies, interest often lies in the analysis of the interevent, or gap times and the association between event times. Gap time analyses are challenging however, even when the length of follow-up is determined independently of the event process, because associations between gap times induce dependent censoring for second and subsequent gap times. This article discusses nonparametric estimation of the association between consecutive gap times based on Kendall's τ in the presence of this type of dependent censoring. A nonparametric estimator that uses inverse probability of censoring weights is provided. Estimates of conditional gap time distributions can be obtained following specification of a particular copula function. Simulation studies show the estimator performs well and compares favorably with an alternative estimator. Generalizations to a piecewise constant Clayton copula are given. Several simulation studies and illustrations with real data sets are also provided.


Agreement between functional residual capacity estimated via automated gas dilution versus via computed tomography in a pleural effusion model.
The measurement of functional residual capacity (FRC) in ventilated patients could help track the extent of acute lung disease, monitor recruitment of unstable lung units, or guide the use of PEEP. Quantitative analysis of computed tomography (CT) images of the lungs is currently the accepted standard for FRC measurement (FRC-CT), but is impractical for routine use. Gas dilution and gas tracer technologies, while attractive for research applications, require specialized equipment and skills missing from the clinical setting. We simultaneously evaluated FRC-CT and FRC determined by a ventilator-incorporated wash-in/wash-out (FRC-WI/WO) method in an animal model of unilateral pleural effusion that varied the fluid volume instilled and the applied PEEP. A swine model (n = 6) of unilateral pleural effusion was created by injecting boluses of radio-opaque fluid (iopromide) (13 mL/kg and then 26 mL/kg) into the right thoracic cavity. FRC-CT and FRC-WI/WO were simultaneously obtained, at 2 PEEP levels, at baseline and at both pleural-effusion volumes. A correlation coefficient (r²) of 0.89 between FRC-CT and FRC-WI/WO revealed concordance between the techniques, with directional agreement and acceptable bias and precision under all tested conditions. We found excellent concordance between FRC-WI/WO and FRC-CT in an animal model of unilateral pleural effusion that stressed the capability of this technology. The technical advantage of the wash-in/wash-out technique is its incorporation into ventilator operation without requiring adjustments to ventilation.


A role for the actin cytoskeleton in the hormonal and growth-factor-mediated activation of protein kinase B.
We show here that cytochalasin D-induced depolymerization of actin filaments markedly reduces the stimulus-dependent activation of protein kinase B (PKB) in four different cell types (HEK-293 cells, L6 myotubes, 3T3-L1 adipocytes and U87MG cells). HEK-293 cells expressing the pleckstrin homology (PH) domains of PKB and general receptor for phosphoinositides-1 (GRP1) fused to green fluorescent protein (GFP) were used to monitor production of 3-phosphoinositides in the plasma membrane. Disassembly of the actin cytoskeleton significantly reduced the insulin-mediated translocation of both PKB-PH-GFP and GRP1-PH-GFP to the plasma membrane, consistent with diminished synthesis of 3-phosphoinositides. Actin depolymerization did not affect the hormonal activation of phosphoinositide 3-kinase (PI 3-kinase), and since cytochalasin D treatment also led to reduced platelet-derived growth factor (PDGF)-induced phosphorylation of PKB in U87MG cells, a PTEN (phosphatase and tensin homologue deleted on chromosome 10) null cell line, lipid phosphatase activity was unlikely to account for any reduction in cellular 3-phosphoinositides. Withdrawal of cytochalasin D from the extracellular medium induced actin filament repolymerization, and reinstated both the recruitment of PH-GFP fusion proteins to the plasma membrane and PKB activation in response to insulin and PDGF. Our findings indicate that an intact actin network is a crucial requirement for PI 3-kinase-mediated production of 3-phosphoinositides and, therefore, for the activation of PKB.


A Comparison of the Effects of the GLP-1 Analogue Liraglutide and Insulin Glargine on Endothelial Function and Metabolic Parameters: A Randomized, Controlled Trial Sapporo Athero-Incretin Study 2 (SAIS2).
GLP-1 improves hyperglycemia, and it has been reported to have favorable effects on atherosclerosis. However, it has not been fully elucidated whether GLP-1 is able to improve endothelial function in patients with type 2 diabetes. Therefore, we investigated the efficacy of the GLP-1 analogue, liraglutide on endothelial function and glycemic metabolism compared with insulin glargine therapy. In this multicenter, prospective randomized parallel-group comparison study, 31 diabetic outpatients (aged 60.3 ± 10.3 years with HbA1c levels of 8.6 ± 0.8%) with current metformin and/or sulfonylurea treatment were enrolled and randomly assigned to receive liraglutide or glargine therapy once daily for 14 weeks. Flow mediated dilation (FMD), a comprehensive panel of hemodynamic parameters (Task Force Monitor), and serum metabolic markers were assessed before and after the treatment period. A greater reduction (worsening) in %FMD was observed in the glargine group, although this change was not statistically different from the liraglutide group (liraglutide; 5.7 to 5.4%, glargine 6.7 to 5.7%). The augmentation index, C-peptide index, derivatives of reactive oxygen metabolites and BMI were significantly improved in the liraglutide group. Central systolic blood pressure and NT-proBNP also tended to be improved in the liraglutide-treated group, while improvements in HbA1c levels were similar between groups. Cardiac index, blood pressure and most other metabolic parameters were not different. Regardless of glycemic improvement, early liraglutide therapy did not affect endothelial function but may provide favorable effects on beta-cell function and cardioprotection in type 2 diabetics without advanced atherosclerosis. UMIN Clinical Trials Registry System as trial ID UMIN000005331.


Successful resection of sigmoid colon cancer in a patient with factor XI deficiency.
A 42-year-old-women with sigmoid colon adenocarcinoma was found to have isolated prolonged activated partial thromboplastin time (aPTT 102.5 s, normal range 24-36 s) preoperatively. Her medical history included an episode of prolonged postdelivery uterine bleeding 16 years previously. A mixed aPTT test showed immediate correction of the prolonged aPTT, indicating a coagulation factor deficiency in the intrinsic pathway. Factor assays showed factor XI was below 1% of average normal value whereas factor VIII, IX and XII activities were normal. Family screening revealed one sister among the three siblings also had isolated prolonged aPTT. The patient was transfused with four units (5mL/kg) of fresh frozen plasma the day before surgery, then with two units during surgery. The operation was uneventful with no bleeding problems. The patient recovered smoothly and is currently undergoing adjuvant chemotherapy. This is the first formal report of a patient with factor XI deficiency undergoing major surgery in Taiwan. Careful monitoring of aPTT, with fresh frozen plasma transfusion, when needed, may safely overcome bleeding problems during surgery.


Physiological and Biochemical Effects of Intrinsically High and Low Exercise Capacities Through Multiomics Approaches.
Regular exercise prevents lipid abnormalities and conditions such as diabetes mellitus, hypertension, and obesity; it considerably benefits sedentary individuals. However, individuals exhibit highly variable responses to exercise, probably due to genetic variations. Animal models are typically used to investigate the relationship of intrinsic exercise capacity with physiological, pathological, psychological, behavioral, and metabolic disorders. In the present study, we investigated differential physiological adaptations caused by intrinsic exercise capacity and explored the regulatory molecules or mechanisms through multiomics approaches. Outbred ICR mice (n = 100) performed an exhaustive swimming test and were ranked based on the exhaustive swimming time to distinguish intrinsically high- and low-capacity groups. Exercise performance, exercise fatigue indexes, glucose tolerance, and body compositions were assessed during the experimental processes. Furthermore, the gut microbiota, transcriptome, and proteome of soleus muscle with intrinsically high exercise capacity (HEC) and low exercise capacity (LEC) were further analyzed to reveal the most influential factors associated with differential exercise capacities. HEC mice outperformed LEC mice in physical activities (exhaustive swimming and forelimb grip strength tests) and exhibited higher glucose tolerance than LEC mice. Exercise-induced peripheral fatigue and the level of injury biomarkers (lactate, ammonia, creatine kinase, and aspartate aminotransferase) were also significantly lower in HEC mice than in LEC mice. Furthermore, the gut of the HEC mice contained significantly more Butyricicoccus than that of the LEC mice. In addition, transcriptome data of the soleus muscle revealed that the expression of microRNAs that are strongly associated with exercise performance-related physiological and metabolic functions (i.e., miR-383, miR-107, miR-30b, miR-669m, miR-191, miR-218, and miR-224) was higher in HEC mice than in LEC mice. The functional proteome data of soleus muscle indicated that the levels of key proteins related to muscle function and carbohydrate metabolism were also significantly higher in HEC mice than in LEC mice. Our study demonstrated that the mice with various intrinsic exercise capacities have different gut microbiome as well as transcriptome and proteome of soleus muscle by using multiomics approaches. The specific bacteria and regulatory factors, including miRNA and functional proteins, may be highly correlated with the adaptation of physiological functions and exercise capacity.