Utilization of human respiratory secretions by mucoid Pseudomonas aeruginosa of cystic fibrosis origin.
Growth and exoproduct production were examined with sputum from patients with respiratory diseases serving as the growth substrate for mucoid strains of Pseudomonas aeruginosa isolated from cystic fibrosis (CF) patients. Mucoid strains are uniquely common to chronic respiratory infections of CF patients. The mucoid colonial morphology of P. aeruginosa is due to the biosynthesis of the exopolysaccharide alginate. Alginate-producing (Alg+) strains utilized CF sputum for growth and high yields of alginate; however, sputum from patients with other respiratory diseases produced comparable results. Analysis of CF sputum medium indicated that amino acids and small peptides were major substrates for P. aeruginosa in respiratory secretions. Cultures of Alg+ strains in CF sputum medium were inhibited in growth and reduced in alginate yields by a low concentration (1 mM) of D-mannose, suggesting therapeutic applications. The rates of growth of two Alg+ strains in CF sputum medium were found to be slightly lower compared with their respective spontaneous Alg- mutants, indicating that the mucoid phenotype does not enhance the ability of P. aeruginosa to utilize respiratory secretions. At all stages of growth in CF sputum medium, two Alg+ strains produced lower yields of protease than did their respective Alg- mutants. When seven Alg+ strains of CF origin were compared with their respective Alg- mutants, the Alg+ phenotype correlated with reduced yields of extracellular proteases. These data are consistent with the hypothesis that mucoid strains of P. aeruginosa are more suited to chronic rather than to acute respiratory infections in that reduced yields of proteases temper the level of damage to the lungs and result in a reduced infiltration of phagocytic cells.


Genome-wide double-stranded RNA sequencing reveals the functional significance of base-paired RNAs in Arabidopsis.
The functional structure of all biologically active molecules is dependent on intra- and inter-molecular interactions. This is especially evident for RNA molecules whose functionality, maturation, and regulation require formation of correct secondary structure through encoded base-pairing interactions. Unfortunately, intra- and inter-molecular base-pairing information is lacking for most RNAs. Here, we marry classical nuclease-based structure mapping techniques with high-throughput sequencing technology to interrogate all base-paired RNA in Arabidopsis thaliana and identify ∼200 new small (sm)RNA-producing substrates of RNA-DEPENDENT RNA POLYMERASE6. Our comprehensive analysis of paired RNAs reveals conserved functionality within introns and both 5' and 3' untranslated regions (UTRs) of mRNAs, as well as a novel population of functional RNAs, many of which are the precursors of smRNAs. Finally, we identify intra-molecular base-pairing interactions to produce a genome-wide collection of RNA secondary structure models. Although our methodology reveals the pairing status of RNA molecules in the absence of cellular proteins, previous studies have demonstrated that structural information obtained for RNAs in solution accurately reflects their structure in ribonucleoprotein complexes. Furthermore, our identification of RNA-DEPENDENT RNA POLYMERASE6 substrates and conserved functional RNA domains within introns and both 5' and 3' untranslated regions (UTRs) of mRNAs using this approach strongly suggests that RNA molecules are correctly folded into their secondary structure in solution. Overall, our findings highlight the importance of base-paired RNAs in eukaryotes and present an approach that should be widely applicable for the analysis of this key structural feature of RNA.


Adherence to physical exercise recommendations in people over 65--the SNAC-Kungsholmen study.
There is limited knowledge regarding to what extent the older population meet the recommendations of physical exercise, especially fitness-enhancing exercise. This study assessed participation in health- and fitness-enhancing exercises in people aged >65, and explored to what extent the possible differences in meeting current recommendations differs by age, gender and education. The study population was derived from the Swedish National study on Aging and Care, and consisted of a random sample of 2593 subjects, aged 65+ years. Participation in health- and fitness-enhancing exercise according to the WHO and the American College of Sports Medicine's recommendations in relation to age, gender and education was evaluated using multinomial logistic regression adjusted for health indicators and physical performance. According to the recommendations, 46% of the participants fulfilled the criteria for health-enhancing and 16% for fitness-enhancing exercises. Independent of health indicators and physical performance, women <80 years of age were less likely than men to participate in fitness-enhancing exercise, but they participated more in health-enhancing exercise. In the advanced age group (80+ years), women were less likely to participate both in fitness- and health-enhancing exercise. Advanced age and low education were negatively related to participation in both health- and fitness-enhancing exercise independent of health indicators, but the association was not observed among people with fast walking speed. Promoting physical exercise and encouraging participation among older adults with lower education, especially among those with initial functional decline, may help to reduce adverse health outcomes.


Induction of apoptosis by quercetin: involvement of heat shock protein.
Quercetin, a widely distributed bioflavonoid, inhibits the growth of tumor cells. The present study was designed to investigate the possible involvement of apoptosis and heat shock protein in the antitumor activity of quercetin. Treatment with quercetin of K562, Molt-4, Raji, and MCAS tumor cell lines resulted in morphological changes, including propidium iodide-stained condensed nuclei (intact or fragmented), condensation of nuclear chromatin, and nuclear fragmentation. Agarose gel electrophoresis of quercetin-treated tumor cells demonstrated a typical ladder-like pattern of DNA fragments. In addition, the hypodiploid DNA peak of propidium iodide-stained nuclei was revealed by flow cytometry. Quercetin induced apoptosis in cells at G1 and S in a dose- and time-dependent manner. The apoptosis-inducing activity of quercetin was enhanced by cycloheximide and actinomycin D. A nuclease inhibitor, aurintricarboxylic acid, inhibited quercetin-induced apoptosis, whereas deprivation of intracellular calcium by EGTA had no effect. 12-O-Tetradecanoylphorbol-13-acetate and H-7 did not affect the induction of apoptosis by quercetin. The synthesis of HSP70 was inhibited by quercetin when determined by immunocytochemistry, Western blot analysis, and Northern blot analysis. Quercetin-treated tumor cells were not induced to show aggregation of HSP70 in the nuclei and nucleolus in response to heat shock, resulting in apoptosis. By contrast, when tumor cells were first exposed to heat shock, no apoptosis was induced by quercetin. In addition, pretreatment of tumor cells with HSP70 antisense oligomer that specifically inhibited the synthesis of HSP70 enhanced the subsequent induction of apoptosis by quercetin. These results suggest that quercetin displays antitumor activity by triggering apoptosis and that HSP70 may affect quercetin-induced apoptosis.


An information-theoretic criterion for intrasubject alignment of FMRI time series: motion corrected independent component analysis.
A three-dimensional image registration method for motion correction of functional magnetic resonance imaging (fMRI) time-series, based on independent component analysis (ICA), is described. We argue that movement during fMRI data acquisition results in a simultaneous increase in the joint entropy of the observed time-series and a decrease in the joint entropy of a nonlinear function of the derived spatially independent components calculated by ICA. We propose this entropy difference as a reliable criterion for motion correction and refer to a method that maximizes this as motion-corrected ICA (MCICA). Specifically, a given motion-corrupted volume may be corrected by determining the linear combination of spatial transformations of the motion-corrupted volume that maximizes the proposed criterion. In essence, MCICA consists of designing an adaptive spatial resampling filter which maintains maximum temporal independence among the recovered components. In contrast with conventional registration methods, MCICA does not require registration of motion-corrupted volumes to a single reference volume which can introduce artifacts because corrections are applied without accounting for variability due to the task-related activation. Simulations demonstrate that MCICA is robust to activation level, additive noise, random motion in the reference volumes and the exact number of independent components extracted. When the method was applied to real data with minimal estimated motion, the method had little effect and, hence, did not introduce spurious changes in the data. However, in a data series from a motor fMRI experiment with larger motion, preprocessing the data with the proposed method resulted in the emergence of activation in primary motor and supplementary motor cortices. Although mutual information (MI) was not explicitly optimized, the MI between all subsequent volumes and the first one was consistently increased for all volumes after preprocessing the data with MCICA. We suggest MCICA represents a robust and reliable method for preprocessing of fMRI time-series corrupted with motion.


Comparative genomics to explore phylogenetic relationship, cryptic sexual potential and host specificity of Rhynchosporium species on grasses.
The Rhynchosporium species complex consists of hemibiotrophic fungal pathogens specialized to different sweet grass species including the cereal crops barley and rye. A sexual stage has not been described, but several lines of evidence suggest the occurrence of sexual reproduction. Therefore, a comparative genomics approach was carried out to disclose the evolutionary relationship of the species and to identify genes demonstrating the potential for a sexual cycle. Furthermore, due to the evolutionary very young age of the five species currently known, this genus appears to be well-suited to address the question at the molecular level of how pathogenic fungi adapt to their hosts. The genomes of the different Rhynchosporium species were sequenced, assembled and annotated using ab initio gene predictors trained on several fungal genomes as well as on Rhynchosporium expressed sequence tags. Structures of the rDNA regions and genome-wide single nucleotide polymorphisms provided a hypothesis for intra-genus evolution. Homology screening detected core meiotic genes along with most genes crucial for sexual recombination in ascomycete fungi. In addition, a large number of cell wall-degrading enzymes that is characteristic for hemibiotrophic and necrotrophic fungi infecting monocotyledonous hosts were found. Furthermore, the Rhynchosporium genomes carry a repertoire of genes coding for polyketide synthases and non-ribosomal peptide synthetases. Several of these genes are missing from the genome of the closest sequenced relative, the poplar pathogen Marssonina brunnea, and are possibly involved in adaptation to the grass hosts. Most importantly, six species-specific genes coding for protein effectors were identified in R. commune. Their deletion yielded mutants that grew more vigorously in planta than the wild type. Both cryptic sexuality and secondary metabolites may have contributed to host adaptation. Most importantly, however, the growth-retarding activity of the species-specific effectors suggests that host adaptation of R. commune aims at extending the biotrophic stage at the expense of the necrotrophic stage of pathogenesis. Like other apoplastic fungi Rhynchosporium colonizes the intercellular matrix of host leaves relatively slowly without causing symptoms, reminiscent of the development of endophytic fungi. Rhynchosporium may therefore become an object for studying the mutualism-parasitism transition.


A cross-sectional validation study of EQ-5D among the Malaysian adult population.
The objective of this study was to assess the construct validity of the EQ-5D instrument among the Malaysian population. This was a cross-sectional study conducted among Malaysian adults in three northern states of Malaysia. A pre-developed questionnaire consisting of both the EQ-5D and SF-12 items was used for data collection. Concurrent, convergent, and known group validity of EQ-5D were assessed against SF-12 and several known relationships with participants' demographic and illness characteristics. A total of 596 Malaysians participated in the study. The mean EQ-5D score was 0.93 (SD = 0.13), while the mean physical component score (PCS-12) and mental component score (MCS-12) scores were 48.9 (SD = 7.4) and 49.1 (SD = 8.0), respectively. Participants with a current medical problem had lower PCS-12 and MCS-12 scores and reported more problems with all of the EQ-5D dimensions; they also had lower EQ-5D and EQ-VAS scores (P < 0.05). Convergent validity was supported by a moderately positive correlation between EQ-5D and EQ-VAS with MCS-12 and PCS-12 scores; moreover, the stronger effect sizes between PCS-12 and the physical dimensions of EQ-5D as well as between MCS-12 with anxiety/depression scores further supported the convergent validity of EQ-5D. Responses to the EQ-5D dimensions only supported two of the four known group validity hypotheses of higher quality of life among individuals who are better educated and no medical problem. No association was found between income and gender with EQ-5D score. This study has demonstrated acceptable construct validity of the EQ-5D among the Malaysian population.


Generation of functional ion-channel tools by E3 targeting.
Here we describe a strategy for generating ion-channel inhibitors. It takes advantage of antibody specificity combined with a pattern recognition approach that targets the third extracellular region (E3) of a channel. To test the concept, we first focused on TRPC5, a member of the transient receptor potential (TRP) calcium channel family, the study of which has been hindered by poor pharmacological tools. Extracellular application of E3-targeted anti-TRPC5 antibody led to a specific TRPC5 inhibitor, enabling TRPC5 to be distinguished from its closest family members, and TRPC5 function to be explored in a relatively intractable physiological system. E3 targeting was further applied to voltage-gated sodium channels, leading to discovery of a subtype-specific inhibitor of Na(V)1.5. These examples illustrate the potential power of E3 targeting as a systematic method for producing gene-type specific ion-channel inhibitors for use in routine assays on cells or tissues from a range of species and having therapeutic potential.


Computational assessment of lipid production in Rhodosporidium toruloides in two-stage and one-stage batch bioprocesses.
Oleaginous yeasts are promising platforms for microbial lipids production as a renewable and sustainable alternative to vegetable oils in biodiesel production. In this paper, a thorough in silico assessment of lipid production in batch cultivation by Rhodosporidium toruloides was developed. By means of dynamic flux balance analysis, the traditional two-stage bioprocess (TSB) performed by the native strain was contrasted with one-stage bioprocess (OSB) using four designed strains obtained by gene knockout strategies. Lipid titer, yield, content, and productivity were analyzed at different initial C/N ratios as relevant performance indicators used in bioprocesses. By weighting these indicators, a global lipid efficiency metric (GLEM) was defined to consider different scenarios. Under simulated conditions, designed strains for lipid overproduction in OSB outperformed the TSB in terms of lipid title (up to threefold), lipid yield (up to 2.4-fold), lipid content (up to 2.8-fold, with a maximum of 76%), and productivity (up to 1.3-fold), depending on C/N ratios. Using these efficiency parameters and the proposed GLEM, the process of selecting the most suitable candidates for lipid production could be carried out before experimental assays. This methodology holds the potential to be extended to other oleaginous microorganisms and diverse strain design techniques.


Dual mode of interaction of DNA polymerase epsilon with proliferating cell nuclear antigen in primer binding and DNA synthesis.
Proliferating cell nuclear antigen can interact with DNA polymerase epsilon on linear DNA templates, even in the absence of other auxiliary factors (replication factor C, replication protein A), and thereby stimulate its primer recognition and DNA synthesis. Using four characterized mutants of proliferating cell nuclear antigen containing three or four alanine residue substitutions on the C-terminal side and the back side of the trimer, we have tested the kinetics of primer binding and nucleotide incorporation by DNA polymerase epsilon in different assays. In contrast with what has been found in interaction studies between DNA polymerase delta and proliferating cell nuclear antigen, our data suggested that stimulation of DNA polymerase epsilon primer binding involves interactions with both the C-terminal side and the back side of proliferating cell nuclear antigen. However, for stimulation of DNA polymerase epsilon DNA synthesis, exclusively the C-terminal side appears to be sufficient. The significance of this dual interaction is discussed with reference to the physiological roles of DNA polymerase epsilon and its interaction with the clamp proliferating cell nuclear antigen.


Construction and analysis of the NCI-EDRN breast cancer reference set for circulating markers of disease.
Many circulating biomarkers have been reported for the diagnosis of breast cancer, but few, if any, have undergone rigorous credentialing using prospective cohorts and blinded evaluation. The NCI Early Detection Research Network (EDRN) has created a prospective, multicenter collection of plasma and serum samples from 832 subjects designed to evaluate circulating biomarkers for the detection and diagnosis of breast cancer. These samples are available to investigators who wish to evaluate their biomarkers using a set of blinded samples. The breast cancer reference set is composed of blood samples collected using a standard operating procedure at four U.S. medical centers from 2008 to 2010 from women undergoing either tissue diagnosis for breast cancer or routine screening mammography. The reference set contains samples from women with incident invasive cancer (n = 190), carcinoma in situ (n = 55), benign pathology with atypia (n = 63), benign disease with no atypia (n = 231), and women with no evidence of breast disease by screening mammography (BI-RADS 1 or 2, n = 276). Using a subset of plasma samples (n = 505) from the reference set, we analyzed 90 proteins by multiplexed immunoassays for their potential utility as diagnostic markers. We found that none of these markers is useful for distinguishing cancer from benign controls. However, elevated CA-125 does appear to be a candidate marker for estrogen receptor-negative cancers. Markers that can distinguish benign breast conditions from invasive cancer have not yet been found. Availability of prospectively collected samples should improve future validation efforts.


Highly Loaded Cellulose/Poly (butylene succinate) Sustainable Composites for Woody-Like Advanced Materials Application.
We report the manufacturing and characterization of poly (butylene succinate) (PBS) and micro cellulose (MCC) woody-like composites. These composites can be applied as a sustainable woody-like composite alternative to conventional fossil polymer-based wood-plastic composites (WPC). The PBS/MCC composites were prepared by using a melt blending of 70 wt% of MCC processed from bleached softwood. MCC was modified to enhance dispersion and compatibility by way of carbodiimide (CDI), polyhydroxy amides (PHA), alkyl ester (EST), (3-Aminopropyl) trimethoxysilane (APTMS), maleic acid anhydride (MAH), and polymeric diphenylmethane diisocyanate (PMDI). The addition of filler into PBS led to a 4.5-fold improvement of Young's modulus E for the MCC composite, in comparison to neat PBS. The 1.6-fold increase of E was obtained for CDI modified composition in comparison to the unmodified MCC composite. At room temperature, the storage modulus E' was found to improve by almost 4-fold for the APTMS composite. The EST composite showed a pronounced enhancement in viscoelasticity properties due to the introduction of flexible long alkyl chains in comparison to other compositions. The glass transition temperature was directly affected by the composition and its value was -15 °C for PBS, -30 °C for EST, and -10 °C for MAH composites. FTIR indicated the generation of strong bonding between the polymer and cellulose components in the composite. Scanning electron microscopy analysis evidenced the agglomeration of the MCC in the PBS/MCC composites. PMDI, APTMS, and CDI composites were characterized by the uniform dispersion of MCC particles and a decrease of polymer crystallinity. MCC chemical modification induced the enhancement of the thermal stability of MCC composites.


Highly multiplexed single-cell in situ RNA and DNA analysis with bioorthogonal cleavable fluorescent oligonucleotides.
The ability to profile transcripts and genomic loci comprehensively in single cells in situ is essential to advance our understanding of normal physiology and disease pathogenesis. Here we report a highly multiplexed single-cell in situ RNA and DNA analysis approach using bioorthogonal cleavable fluorescent oligonucleotides. In this approach, oligonucleotides tethered to fluorophores through an azide-based cleavable linker are used to detect their nucleic acids targets by in situ hybridization. After fluorescence imaging, the fluorophores in the whole specimen are efficiently cleaved in 30 minutes without loss of RNA or DNA integrity. Through reiterative cycles of hybridization, imaging, and cleavage, this method has the potential to quantify hundreds to thousands of different RNA species or genomic loci in single cells in situ at the single-molecule sensitivity. Applying this approach, we demonstrate that different nucleic acids can be detected in each hybridization cycle by multi-color staining, and at least ten continuous hybridization cycles can be carried out in the same specimen. We also show that the integrated single-cell in situ analysis of DNA, RNA and protein can be achieved using cleavable fluorescent oligonucleotides combined with cleavable fluorescent antibodies. This highly multiplexed imaging platform will have wide applications in systems biology and biomedical research.


DART: deep learning enabled topological interaction model for energy prediction of metal clusters and its application in identifying unique low energy isomers.
Recently, machine learning (ML) has proven to yield fast and accurate predictions of chemical properties to accelerate the discovery of novel molecules and materials. The majority of the work is on organic molecules, and much more work needs to be done for inorganic molecules, especially clusters. In the present work, we introduce a simple topological atomic descriptor called TAD, which encodes chemical environment information of each atom in the cluster. TAD is a simple and interpretable descriptor where each value represents the atom count in three shells. We also introduce the DART deep learning enabled topological interaction model, which uses TAD as a feature vector to predict energies of metal clusters, in our case gallium clusters with sizes ranging from 31 to 70 atoms. The DART model is designed based on the principle that the energy is a function of atomic interactions and allows us to model these complex atomic interactions to predict the energy. We further introduce a new dataset called GNC_31-70, which comprises structures and DFT optimized energies of gallium clusters with sizes ranging from 31 to 70 atoms. We show how DART can be used to accelerate the process of identification of low energy structures without geometry optimization. Albeit using a topological descriptor, DART achieves a mean absolute error (MAE) of 3.59 kcal mol-1 (0.15 eV) on the test set. We also show that our model can distinguish core and surface atoms in the Ga-70 cluster, which the model has never encountered earlier. Finally, we demonstrate the transferability of the DART model by predicting energies for about 6k unseen configurations picked up from molecular dynamics (MD) data for three cluster sizes (46, 57, and 60) within seconds. The DART model was able to reduce the load on DFT optimizations while identifying unique low energy structures from MD data.


[Neonatal mortality, analysis of surveillance registers, and neonatal medical histories of 2011 in Huanuco and Ucayali, Peru].
To estimate the rate of neonatal mortality and to describe neonatal deaths in 2011 in hospitals of the Ministry of Health at Huanuco and Ucayali (Peru). Cross-sectional study from September to November 2012 in Huanuco and Ucayali. Records of neonatal deaths in 2011 were reviewed from provincial municipalities, regional health directorates and four referral hospitals. To calculate mortality rates, we used the most reliable information sources by region. Reviewing 185 medical records in hospitals allowed us to describe the root causes of neonatal deaths. In 2011, 10,886 live births and 158 neonatal deaths were reported in Huanuco, with a rate of 14.5 deaths per 1000 live births. In Ucayali, 11,441 live births and 138 neonatal deaths were reported, with a rate of 12.1 deaths per 1000 live births. Most hospital neonatal deaths occurred in the first 7 days of life (87%), in preterm infants (73.9%) and with low birth weight (67%). The most common underlying causes of neonatal deaths were infection (31.4%), congenital malformation (22.2%) and prematurity (18.9%). Neonatal mortality rate in the studied regions was higher than the national average. The results suggest the need for effective and comprehensive interventions during pregnancy, childbirth and the early postnatal period; this last period is the most vulnerable in the neonate.


Risk of secondary progressive multiple sclerosis: A longitudinal study.
The risk factors for conversion from relapsing-remitting to secondary progressive multiple sclerosis remain highly contested. The aim of this study was to determine the demographic, clinical and paraclinical features that influence the risk of conversion to secondary progressive multiple sclerosis. Patients with adult-onset relapsing-remitting multiple sclerosis and at least four recorded disability scores were selected from MSBase, a global observational cohort. The risk of conversion to objectively defined secondary progressive multiple sclerosis was evaluated at multiple time points per patient using multivariable marginal Cox regression models. Sensitivity analyses were performed. A total of 15,717 patients were included in the primary analysis. Older age (hazard ratio (HR) = 1.02, p < 0.001), longer disease duration (HR = 1.01, p = 0.038), a higher Expanded Disability Status Scale score (HR = 1.30, p < 0.001), more rapid disability trajectory (HR = 2.82, p < 0.001) and greater number of relapses in the previous year (HR = 1.07, p = 0.010) were independently associated with an increased risk of secondary progressive multiple sclerosis. Improving disability (HR = 0.62, p = 0.039) and disease-modifying therapy exposure (HR = 0.71, p = 0.007) were associated with a lower risk. Recent cerebral magnetic resonance imaging activity, evidence of spinal cord lesions and oligoclonal bands in the cerebrospinal fluid were not associated with the risk of conversion. Risk of secondary progressive multiple sclerosis increases with age, duration of illness and worsening disability and decreases with improving disability. Therapy may delay the onset of secondary progression.


Interactions of human peritoneal mesothelial cells with serous ovarian cancer cell spheroids--evidence for a mechanical and paracrine barrier function of the peritoneal mesothelium.
Ovarian carcinoma spreads by implantation of tumor cells onto the peritoneal mesothelium. We established a 3-dimensional coculture model to simulate the interactions of ovarian carcinoma cell aggregates with human peritoneal mesothelial cells (HPMC). Multicellular tumor spheroids (MCTS) of the human ovarian cancer cell line SK-OV-3 were directly inoculated onto either confluent HPMC monolayers or their submesothelial matrix or were cocultured with mesothelium without direct cellular contact. Inoculation of MCTS onto submesothelial matrix resulted in rapid attachment (within 30 minutes) of the tumor cell aggregates followed by rapid dissemination (within 12 hours) and growth of tumor cells. Intact mesothelium increased the time required for MCTS attachment (up to 180 minutes) and led to almost complete inhibition of tumor cell dissemination and to 47% tumor growth suppression. Bromodeoxyuridine incorporation into tumor cell nuclei was almost completely abolished in cocultured MCTS. Growth also was inhibited in MCTS treated with supernatants of HPMC. Analysis of coculture supernatants revealed that HPMC-derived transforming growth factor β (TGF-β) was almost completely bound by MCTS. Addition of a function-blocking anti-TGF-β antibody (30 μg/mL) to the cocultures abrogated the growth inhibitory effect of the mesothelium by 50%. The present model provides a dynamic system to study the complex interactions of ovarian carcinoma cells with HPMC over extended periods and suggests that the mesothelium constitutes a mechanical and partly TGF-β-mediated paracrine barrier to the progression of ovarian cancer.


An Assessment of Osteoporotic Conditions among Users and Non-Users of Warfarin: A Case-Control Study.
Warfarin is an oral anticoagulant which is associated with a decrease in Bone Mass Density (BMD). The study aimed to explore the prevalence of osteoporosis conditions in Malaysians with chronic warfarin patients using calcaneal quarter of Quantitative Ultrasound (QUS) machine and to investigate whether long-term sodium warfarin therapy that antagonizes vitamin K is affecting the increasing rate of osteoporosis in Penang and to find the risk factors of getting osteoporosis among warfarin users and its non users. A case-control study was conducted among 130 patients using warfarin, attending the outpatient clinic at Hospital Pulau Pinang. A convenience sampling method was used to recruit the required sample. Another 140 subjects were selected from the community as a control group (non-users of warfarin). This study showed that more than three-quarter patients (82%) were at high risk of abnormal BMD. The warfarin users were two times more likely to have a higher osteoporosis risk compared to control group. Moreover, BMD has a negative correlation with age, but has a positive correlation with Body Mass Index (BMI). Patients showed a negative correlation with a higher dose of warfarin intake. This study concluded that osteopenia and osteoporosis are serious problems between users and non-users warfarin in Penang. Therefore, it should be taken into consideration in the Malaysian Health Ministry's agenda.


A preliminary study of functional connectivity in comorbid adolescent depression.
Major depressive disorder (MDD) begins frequently in adolescence and is associated with severe outcomes, but the developmental neurobiology of MDD is not well understood. Research in adults has implicated fronto-limbic neural networks in the pathophysiology of MDD, particularly in relation to the subgenual anterior cingulate cortex (ACC). Developmental changes in brain networks during adolescence highlight the need to examine MDD-related circuitry in teens separately from adults. Using resting state functional magnetic resonance imaging (fMRI), this study examined functional connectivity in adolescents with MDD (n=12) and healthy adolescents (n=14). Seed-based connectivity analysis revealed that adolescents with MDD have decreased functional connectivity in a subgenual ACC-based neural network that includes the supragenual ACC (BA 32), the right medial frontal cortex (BA 10), the left inferior (BA 47) and superior frontal cortex (BA 22), superior temporal gyrus (BA 22), and the insular cortex (BA 13). These preliminary data suggest that MDD in adolescence is associated with abnormal connectivity within neural circuits that mediate emotion processing. Future research in larger, un-medicated samples will be necessary to confirm this finding. We conclude that hypothesis-driven, seed-based analyses of resting state fMRI data hold promise for advancing our current understanding of abnormal development of neural circuitry in adolescents with MDD.


Effect of Finishing Techniques on the Marginal Integrity of Resin-Based Composite and Resin-Modified Glass Ionomer Restoration.
The purpose of this study was to evaluate the marginal integrity of resin-based composite (RBC and resin-modified glass ionomer cement (RMGIC) restorations as a function of finishing technique and location of the tooth. Forty extracted third molars were assigned to four groups (N = 10) according to finishing instruments (aluminum oxide discs, fluted carbides, fine diamonds, and coarse diamond). Each specimen received standardized Class V preparations on the facial and lingual surfaces with occlusal margins on enamel and gingival margins on dentin. Each preparation was randomly assigned to be restored with either RBC or RMGIC. Specimens were finished with standardized pressure at approximately 0.16 N and evaluated at a magnification of 600× using an environmental scanning electron microscope. Occlusal and gingival margins were analyzed using an imaging software, and means for all measured gaps were calculated. Data were analyzed with a factorial analysis of variance. All possible two-way interactions were included, and the level of significance was set at 0.05. There were no statistically significant differences among the four types of finishing instruments used in the study. RBC-restored specimens exhibited significantly smaller mean marginal gaps (1.70-7.56 μm) than RMGI-restored specimens (5.24-14.24 μm) in enamel and dentin margins, respectively. There was a statistically significant difference between enamel and dentin with regard to marginal gap formation. Under the conditions of this study, marginal gap formation was not affected by finishing technique. RBC margins exhibited significantly less marginal gap than RMGI margins, whereas enamel margins resulted in significantly less marginal gap than did dentin margins. Multiple factors can affect the marginal integrity and the longevity of direct restorations. From these, the finishing and polishing techniques are critical steps that are under the clinician's control, and proper finishing and polishing techniques should be applied for avoiding introduction of stress to the margin of the restoration. It seems that instrumentation do not play a significant role, as much as the restorative material and the substrate to bond.


Progressive degeneration of human neural stem cells caused by pathogenic LRRK2.
Nuclear-architecture defects have been shown to correlate with the manifestation of a number of human diseases as well as ageing. It is therefore plausible that diseases whose manifestations correlate with ageing might be connected to the appearance of nuclear aberrations over time. We decided to evaluate nuclear organization in the context of ageing-associated disorders by focusing on a leucine-rich repeat kinase 2 (LRRK2) dominant mutation (G2019S; glycine-to-serine substitution at amino acid 2019), which is associated with familial and sporadic Parkinson's disease as well as impairment of adult neurogenesis in mice. Here we report on the generation of induced pluripotent stem cells (iPSCs) derived from Parkinson's disease patients and the implications of LRRK2(G2019S) mutation in human neural-stem-cell (NSC) populations. Mutant NSCs showed increased susceptibility to proteasomal stress as well as passage-dependent deficiencies in nuclear-envelope organization, clonal expansion and neuronal differentiation. Disease phenotypes were rescued by targeted correction of the LRRK2(G2019S) mutation with its wild-type counterpart in Parkinson's disease iPSCs and were recapitulated after targeted knock-in of the LRRK2(G2019S) mutation in human embryonic stem cells. Analysis of human brain tissue showed nuclear-envelope impairment in clinically diagnosed Parkinson's disease patients. Together, our results identify the nucleus as a previously unknown cellular organelle in Parkinson's disease pathology and may help to open new avenues for Parkinson's disease diagnoses as well as for the potential development of therapeutics targeting this fundamental cell structure.


Evolution of neo-sex chromosomes in Silene diclinis.
A small cluster of dioecious species in the plant genus Silene has evolved chromosomal sex determination and sex chromosomes relatively recently, within the last 10 million years (MY). Five dioecious Silene species (section Elisanthe) are very closely related (1-2 MY of divergence) and it was previously thought that all five have similar sex chromosomes. Here we demonstrate that in one of these species, Silene diclinis, the sex chromosomes have been significantly rearranged, resulting in the formation of neo-sex chromosomes. Fluorescence in situ hybridization with genic and repetitive probes revealed that in S. diclinis a reciprocal translocation has occurred between the ancestral Y chromosome and an autosome, resulting in chromosomes designated Y1 and Y2. Both Y1 and Y2 chromosomes are male specific. Y1 pairs with the X chromosome and with the autosome (the neo-X), which cosegregates with X. Y2 pairs only with the neo-X, forming a chain X-Y1-neo-X-Y2 in male meiosis. Despite very recent formation of the neo-sex chromosomes in S. diclinis, they are present in all surveyed individuals throughout the species range. Evolution of neo-sex chromosomes may be the cause of partial reproductive isolation of this species and could have been the isolating mechanism that drove speciation of S. diclinis.


High-quality genome (re)assembly using chromosomal contact data.
Closing gaps in draft genome assemblies can be costly and time-consuming, and published genomes are therefore often left 'unfinished.' Here we show that genome-wide chromosome conformation capture (3C) data can be used to overcome these limitations, and present a computational approach rooted in polymer physics that determines the most likely genome structure using chromosomal contact data. This algorithm--named GRAAL--generates high-quality assemblies of genomes in which repeated and duplicated regions are accurately represented and offers a direct probabilistic interpretation of the computed structures. We first validated GRAAL on the reference genome of Saccharomyces cerevisiae, as well as other yeast isolates, where GRAAL recovered both known and unknown complex chromosomal structural variations. We then applied GRAAL to the finishing of the assembly of Trichoderma reesei and obtained a number of contigs congruent with the know karyotype of this species. Finally, we showed that GRAAL can accurately reconstruct human chromosomes from either fragments generated in silico or contigs obtained from de novo assembly. In all these applications, GRAAL compared favourably to recently published programmes implementing related approaches.


Lipopolysaccharide-induced hypotension and vascular hyporeactivity in the rat: tissue analysis of nitric oxide synthase mRNA and protein expression in the presence and absence of dexamethasone, NG-monomethyl-L-arginine or indomethacin.
The role of inducible nitric oxide synthase (iNOS) was examined in the hypotension and vascular hyporesponsiveness to norepinephrine (NE) invoked by lipopolysaccharide (LPS) in pentobarbital-anesthetized rats. Saline, dexamethasone (DEX), NG-monomethyl-L-arginine (LNMMA) or indomethacin (IND) were administered either pre-LPS (0.5 hr) or post-LPS (4.5 hr) treatment. Rats were then challenged with NE 10 min before LPS injection and 1, 4, and 5 hr after LPS. Administration of LPS produced a biphasic hypotension: an immediate hypotension, which partially recovered within 15 min and was unaffected by any of the pretreatments; and a secondary, more prolonged hypotension which was attenuated by DEX, LNMMA and IND. The NE-induced pressor effects were significantly attenuated 1, 4 and 5 hr post LPS. Pretreatment with LNMMA or DEX significantly attenuated the LPS-induced NE hyporesponsiveness 4 and 5 hr post LPS. LNMMA was the only post-LPS treatment able to reverse the NE hyporesponsiveness. The LPS-induced iNOS mRNA and protein expression was demonstrated in the liver, lung, spleen, heart, kidney and brain by Northern hybridization and Western blot analyses. Low levels of neuronal constitutive NOS mRNA and endothelial cell constitutive NOS mRNA were only detected in brain or myocardial tissue, respectively. Significant induction of iNOS mRNA and protein expression was also observed in the liver, lung and spleen of rats pretreated with DEX, LNMMA or IND. The continued expression of iNOS in the presence of a pharmacologically relevant dose of DEX suggests that DEX may not be an optimal pharmacological agent for defining the in vivo roles of iNOS.(ABSTRACT TRUNCATED AT 250 WORDS)


Ethnic differences in the incidence of childhood IDDM in Israel (1965-1993). Marked increase since 1985, especially in Yemenite Jews.
To establish the changes in the incidence of childhood IDDM during the years 1965-1993 in the different ethnic groups in Israel. A whole-country register of childhood IDDM (0-17 years) was started in Israel in 1965. Onset of IDDM was considered to be the date of first insulin injection. The data were collected from all outpatient clinics and hospitals. Ascertainment is estimated to be over 95%. A total of 1,868 patients were registered for a period of 28 years. Marked differences were found between ethnic groups. The highest incidence was among the Yemenite Jews, who reached an incidence of 18.5/10(5), followed by Ashkenazi Jews (10.0/10(5)), non-Ashkenazi Jews, except Yemenites (7.3/10(5)), and Arabs (2.9/10(5)). In addition, it was found that in all Jewish subgroups, in contrast with the Arabs, there was a marked increase in incidence after 1985. Israel is a country with low, intermediate, and high incidence of childhood IDDM. The interethnic differences in incidence are probably due to genetic factors. However, the significant increase in incidence since 1985 in the Jewish population is ascribed to thus far unidentified environmental factors. It is hypothesized that the marked increase in IDDM is due to environmental factors linked to changes in affluence and lifestyle. These may also explain the difference in incidence between the Jewish and Arab populations, the latter living more in rural areas and leading a more traditional lifestyle.


Functional analysis of the superoxide dismutase family in Aspergillus fumigatus.
Reactive oxidant species produced by phagocytes have been reported as being involved in the killing of Aspergillus fumigatus. Fungal superoxide dismutases (SODs) that detoxify superoxide anions could be putative virulence factors for this opportunistic pathogen. Four genes encoding putative Sods have been identified in the A. fumigatus genome: a cytoplasmic Cu/ZnSOD (AfSod1p), a mitochondrial MnSOD (AfSod2p), a cytoplasmic MnSOD (AfSod3p) and AfSod4 displaying a MnSOD C-terminal domain. During growth, AfSOD1 and AfSOD2 were highly expressed in conidia whereas AfSOD3 was only strongly expressed in mycelium. AfSOD4 was weakly expressed compared with other SODs. The deletion of AfSOD4 was lethal. Delta sod1 and Delta sod2 mutants showed a growth inhibition at high temperature and a hypersensitivity to menadione whereas the sod3 mutant had only a slight growth delay at high temperature. Multiple mutations had only an additive effect on the phenotype. The triple sod1/sod2/sod3 mutant was characterized by a delay in conidial germination, a reduced conidial survival during storage overtime, the highest sensitivity to menadione and an increased sensitivity to killing by alveolar macrophage of immunocompetent mice. In spite of these phenotypes, no significant virulence difference was observed between the triple mutant and parental strain in experimental murine aspergillosis models in immunocompromised animals.


Bisphenols and phthalates: Plastic chemical exposures can contribute to adverse cardiovascular health outcomes.
Phthalates and bisphenols are high production volume chemicals that are used in the manufacturing of consumer and medical products. Given the ubiquity of bisphenol and phthalate chemicals in the environment, biomonitoring studies routinely detect these chemicals in 75-90% of the general population. Accumulating evidence suggests that such chemical exposures may influence human health outcomes, including cardiovascular health. These associations are particularly worrisome for sensitive populations, including fetal, infant and pediatric groups-with underdeveloped metabolic capabilities and developing organ systems. In the presented article, we aimed to review the literature on environmental and clinical exposures to bisphenols and phthalates, highlight experimental work that suggests that these chemicals may exert a negative influence on cardiovascular health, and emphasize areas of concern that relate to vulnerable pediatric groups. Gaps in our current knowledge are also discussed, so that future endeavors may resolve the relationship between chemical exposures and the impact on pediatric cardiovascular physiology.


Transcriptome analysis of bitter acid biosynthesis and precursor pathways in hop (Humulus lupulus).
Bitter acids (e.g. humulone) are prenylated polyketides synthesized in lupulin glands of the hop plant (Humulus lupulus) which are important contributors to the bitter flavour and stability of beer. Bitter acids are formed from acyl-CoA precursors derived from branched-chain amino acid (BCAA) degradation and C5 prenyl diphosphates from the methyl-D-erythritol 4-phosphate (MEP) pathway. We used RNA sequencing (RNA-seq) to obtain the transcriptomes of isolated lupulin glands, cones with glands removed and leaves from high α-acid hop cultivars, and analyzed these datasets for genes involved in bitter acid biosynthesis including the supply of major precursors. We also measured the levels of BCAAs, acyl-CoA intermediates, and bitter acids in glands, cones and leaves. Transcripts encoding all the enzymes of BCAA metabolism were significantly more abundant in lupulin glands, indicating that BCAA biosynthesis and subsequent degradation occurs in these specialized cells. Branched-chain acyl-CoAs and bitter acids were present at higher levels in glands compared with leaves and cones. RNA-seq analysis showed the gland-specific expression of the MEP pathway, enzymes of sucrose degradation and several transcription factors that may regulate bitter acid biosynthesis in glands. Two branched-chain aminotransferase (BCAT) enzymes, HlBCAT1 and HlBCAT2, were abundant, with gene expression quantification by RNA-seq and qRT-PCR indicating that HlBCAT1 was specific to glands while HlBCAT2 was present in glands, cones and leaves. Recombinant HlBCAT1 and HlBCAT2 catalyzed forward (biosynthetic) and reverse (catabolic) reactions with similar kinetic parameters. HlBCAT1 is targeted to mitochondria where it likely plays a role in BCAA catabolism. HlBCAT2 is a plastidial enzyme likely involved in BCAA biosynthesis. Phylogenetic analysis of the hop BCATs and those from other plants showed that they group into distinct biosynthetic (plastidial) and catabolic (mitochondrial) clades. Our analysis of the hop transcriptome significantly expands the genomic resources available for this agriculturally-important crop. This study provides evidence for the lupulin gland-specific biosynthesis of BCAAs and prenyl diphosphates to provide precursors for the production of bitter acids. The biosynthetic pathway leading to BCAAs in lupulin glands involves the plastidial enzyme, HlBCAT2. The mitochondrial enzyme HlBCAT1 degrades BCAAs as the first step in the catabolic pathway leading to branched chain-acyl-CoAs.


Implicit visuospatial attention shapes numerosity adaptation and perception.
The perception of numerical quantities is susceptible to adaptation: after inspecting a numerous dot array for a few seconds a subsequent dot array is grossly underestimated. In a recent work we showed that the mere appearance of an additional numerically neutral stimulus significantly reduces the adaptation magnitude. Here we demonstrate that this reduction is likely due to a numerosity underestimation of the adaptor caused by a change of numerosity-related attentional resources deployed on the adapting stimulus. In Experiment 1 we replicated previous findings revealing a robust reduction of numerosity adaptation when an additional adaptor (even if neutral) was displayed. In Experiment 2 we used the method of magnitude estimation to demonstrate that numerosity is underestimated whenever a second task-irrelevant numerical stimulus appears on screen. Furthermore we demonstrated that the same experimental manipulations were not effective in modulating orientation adaptation magnitude as well as orientation estimation accuracy. Our results support the hypothesis of a tight relationship between numerosity perception and implicit visuospatial attention and corroborate the notion that numerosity adaptation depends on perceived rather than physical numerosity. However the lack of an effect of visuospatial attentional deployment for orientation perception suggests that attention might differently shape adaptation aftereffects for different features along the visual hierarchy.


Discovery of a Chemical Tool Inhibitor Targeting the Bromodomains of TRIM24 and BRPF.
TRIM24 is a transcriptional regulator as well as an E3 ubiquitin ligase. It is overexpressed in diverse tumors, and high expression levels have been linked to poor prognosis in breast cancer patients. TRIM24 contains a PHD/bromodomain offering the opportunity to develop protein interaction inhibitors that target this protein interaction module. Here we identified potent acetyl-lysine mimetic benzimidazolones TRIM24 bromodomain inhibitors. The best compound of this series is a selective BRPF1B/TRIM24 dual inhibitor that bound with a KD of 137 and 222 nM, respectively, but exerted good selectivity over other bromodomains. Cellular activity of the inhibitor was demonstrated using FRAP assays as well as cell viability data.


Somitogenesis clock-wave initiation requires differential decay and multiple binding sites for clock protein.
Somitogenesis is a process common to all vertebrate embryos in which repeated blocks of cells arise from the presomitic mesoderm (PSM) to lay a foundational pattern for trunk and tail development. Somites form in the wake of passing waves of periodic gene expression that originate in the tailbud and sweep posteriorly across the PSM. Previous work has suggested that the waves result from a spatiotemporally graded control protein that affects the oscillation rate of clock-gene expression. With a minimally constructed mathematical model, we study the contribution of two control mechanisms to the initial formation of this gene-expression wave. We test four biologically motivated model scenarios with either one or two clock protein transcription binding sites, and with or without differential decay rates for clock protein monomers and dimers. We examine the sensitivity of wave formation with respect to multiple model parameters and robustness to heterogeneity in cell population. We find that only a model with both multiple binding sites and differential decay rates is able to reproduce experimentally observed waveforms. Our results show that the experimentally observed characteristics of somitogenesis wave initiation constrain the underlying genetic control mechanisms.


Cohort Trends in the Burden of Multiple Chronic Conditions Among Aging U.S. Adults.
Multimorbidity, also referred to as multiple chronic conditions (MCCs), is the concurrent presence of 2 or more chronic health conditions. Increasing multimorbidity represents a substantial threat to the health of aging populations. Recent trends suggest greater risk of poor health and mortality among later-born cohorts, yet we are unaware of work examining cohort differences in multimorbidity among aging U.S. adults. We examine intercohort variation in MCC burden in adults aged 51 years and older using 20 years (n = 33,598; 1998-2018) of repeated assessment drawn from the Health and Retirement Study. The index of MCCs included 9 chronic conditions (heart disease, hypertension, stroke, diabetes, arthritis, lung disease, cancer excluding skin cancer, high depressive symptoms, and cognitive impairment). We used linear mixed models with various approaches to estimate age/period/cohort effects to model intercohort patterns in MCC burden. We also explored variation in the specific conditions driving cohort differences in multimorbidity. More recent cohorts had greater MCC burden and developed multimorbidity at earlier ages than those born to prior generations. The burden of chronic conditions was patterned by life-course sociodemographic factors and childhood health for all cohorts. Among adults with multimorbidity, arthritis and hypertension were the most prevalent conditions for all cohorts, and there was evidence that high depressive symptoms and diabetes contributed to the observed cohort differences in multimorbidity risk. Our results suggest increasing multimorbidity burden among more recently born cohorts of aging U.S. adults and should inform policy to address diminishing health in aging populations.


Pathways to homelessness among the mentally ill.
Persons with mental illness are over-represented among the homeless relative to the general population, and mental illness is most likely one of many vulnerabilities that confer risk for homelessness. This paper elucidates the pathways to homelessness for persons with mental illness by comparing and contrasting groups of mentally ill homeless persons, non-mentally ill homeless persons, and housed mentally ill persons drawn from RAND's Course of Homelessness (COH) study and the Epidemiological Catchment Area (ECA) survey. Homeless persons share childhood histories of economic and social disadvantage. The mentally ill homeless appear to have a "double dose" of disadvantage: poverty with the addition of childhood family instability and violence. Among the mentally ill homeless, those who became homeless prior to becoming mentally ill have the highest levels of disadvantage and disruption; while those who become homeless after becoming ill have an especially high prevalence of alcohol dependence. Mental illness may play a role in initiating homelessness for some, but is unlikely in and of itself to be a sufficient risk factor for homelessness. In addition to outreach and treatment programs for adult mentally ill homeless persons, emphasis should be placed on interventions with children and on addressing more pervasive causes of homelessness.


ETV1 is a lineage survival factor that cooperates with KIT in gastrointestinal stromal tumours.
Gastrointestinal stromal tumour (GIST) is the most common human sarcoma and is primarily defined by activating mutations in the KIT or PDGFRA receptor tyrosine kinases. KIT is highly expressed in interstitial cells of Cajal (ICCs)-the presumed cell of origin for GIST-as well as in haematopoietic stem cells, melanocytes, mast cells and germ cells. Yet, families harbouring germline activating KIT mutations and mice with knock-in Kit mutations almost exclusively develop ICC hyperplasia and GIST, suggesting that the cellular context is important for KIT to mediate oncogenesis. Here we show that the ETS family member ETV1 is highly expressed in the subtypes of ICCs sensitive to oncogenic KIT mediated transformation, and is required for their development. In addition, ETV1 is universally highly expressed in GISTs and is required for growth of imatinib-sensitive and resistant GIST cell lines. Transcriptome profiling and global analyses of ETV1-binding sites suggest that ETV1 is a master regulator of an ICC-GIST-specific transcription network mainly through enhancer binding. The ETV1 transcriptional program is further regulated by activated KIT, which prolongs ETV1 protein stability and cooperates with ETV1 to promote tumorigenesis. We propose that GIST arises from ICCs with high levels of endogenous ETV1 expression that, when coupled with an activating KIT mutation, drives an oncogenic ETS transcriptional program. This differs from other ETS-dependent tumours such as prostate cancer, melanoma and Ewing sarcoma where genomic translocation or amplification drives aberrant ETS expression. It also represents a novel mechanism of oncogenic transcription factor activation.


A new study of intraosseous blood for laboratory analysis.
Intraosseous (IO) blood is frequently used to establish a blood chemistry profile in critically ill patients. Questions remain regarding the reliability of IO blood for laboratory analysis and established criteria regarding the amount of marrow/blood to waste before taking an IO sample are not available. To evaluate IO-derived blood for routine laboratory blood tests needed in the care of critically ill patients and to determine the amount of marrow/blood to waste before drawing blood from the IO space for laboratory analysis. Blood samples were drawn from peripheral veins of 10 volunteers. Within 5 minutes, 2 IO blood samples were obtained; one following 2 mL of waste and another following 6 mL of waste. Samples were analyzed for complete blood count and chemistry profile. Values were analyzed using Pearson correlation coefficients. Levels of significance were determined using the t distribution. Mean values for the draws were calculated and compared, with the intravenous blood sample serving as a control for the IO samples. There was a significant correlation between intravenous and IO samples for red blood cell counts and hemoglobin and hematocrit levels but not for white blood cell counts and platelet counts. There was a significant correlation between intravenous and IO samples for glucose, blood urea nitrogen, creatinine, chloride, total protein, and albumin concentrations but not for sodium, potassium, CO(2), and calcium levels. When venous blood cannot be accessed, IO blood aspirate may serve as a reliable alternate, especially for hemoglobin and hematocrit levels and most analytes in a basic blood chemistry profile. Exceptions are CO(2) levels and platelet counts, which may be lower, and white blood cell counts, which may appear elevated.


Using information prescriptions to refer patients with metabolic conditions to the Genetics Home Reference website.
The objectives of this study were to assess the reactions of adult patients and parents of children with metabolic conditions to receipt of an "information prescription" (IP) to visit Genetics Home Reference (GHR), a National Institutes of Health/National Library of Medicine online resource, and evaluate the perceived utility of information found on the site. Patients seen at the University of Utah Metabolic Service Clinic were invited to participate in the study and asked to complete an initial survey to gather demographic data and an online survey six weeks later to obtain information about user experience. Fifty-three of 82 individuals completed both surveys, for an overall response rate of 64.6%. Most respondents (88.7%) agreed that receiving the IP was a "good idea," and nearly all used the IP to visit GHR. More than three-quarters (79.6%) agreed that information on GHR supplemented a physician's advice; 60.4% reported an improved understanding of a health condition; and 41.5% either looked for or would consider looking for additional information. Eighty-six percent of respondents were satisfied with the information found on GHR, and 80% would recommend the site. Use of an IP to direct patients to GHR was well received, and retrieved information was perceived as useful in key areas. The high level of satisfaction with GHR argues for expanded use of the IP approach in this patient population.


Impact of Anesthesia on the Outcome of Acute Ischemic Stroke after Endovascular Treatment with the Solitaire Stent Retriever.
General anesthesia during endovascular treatment of acute ischemic stroke may have an adverse effect on outcome compared with conscious sedation. The aim of this study was to examine the impact of the type of anesthesia on the outcome of patients with acute ischemic stroke treated with the Solitaire stent retriever, accounting for confounding factors. Four-hundred one patients with consecutive acute anterior circulation stroke treated with a Solitaire stent retriever were included in this prospective analysis. Outcome was assessed after 3 months by the modified Rankin Scale. One-hundred thirty-five patients (31%) underwent endovascular treatment with conscious sedation, and 266 patients (69%), with general anesthesia. Patients under general anesthesia had higher NIHSS scores on admission (17 versus 13, P < .001) and more internal carotid artery occlusions (44.6% versus 14.8%, P < .001) than patients under conscious sedation. Other baseline characteristics such as time from symptom onset to the start of endovascular treatment did not differ. Favorable outcome (mRS 0-2) was more frequent with conscious sedation (47.4% versus 32%; OR, 0.773; 95% CI, 0.646-0.925; P = .002) in univariable but not multivariable logistic regression analysis (P = .629). Mortality did not differ (P = .077). Independent predictors of outcome were age (OR, 0.95; 95% CI, 0.933-0.969; P < .001), NIHSS score (OR, 0.894; 95% CI, 0.855-0.933; P < .001), time from symptom onset to the start of endovascular treatment (OR, 0.998; 95% CI, 0.996-0.999; P = .011), diabetes mellitus (OR, 0.544; 95% CI, 0.305-0.927; P = .04), and symptomatic intracerebral hemorrhage (OR, 0.109; 95% CI, 0.028-0.428; P = .002). In this single-center study, the anesthetic management during stent retriever thrombectomy with general anesthesia or conscious sedation had no impact on the outcome of patients with large-vessel occlusion in the anterior circulation.


Side Reactions Do Not Completely Disrupt Linear Self-Replicating Chemical Reaction Systems.
A crucial question within the fields of origins of life and metabolic networks is whether or not a self-replicating chemical reaction system is able to persist in the presence of side reactions. Due to the strong nonlinear effects involved in such systems, they are often difficult to study analytically. There are however certain conditions that allow for a wide range of these reaction systems to be well described by a set of linear ordinary differential equations. In this article, we elucidate these conditions and present a method to construct and solve such equations. For those linear self-replicating systems, we quantitatively find that the growth rate of the system is simply proportional to the sum of all the rate constants of the reactions that constitute the system (but is nontrivially determined by the relative values). We also give quantitative descriptions of how strongly side reactions need to be coupled with the system in order to completely disrupt the system.


Prophylactic radiotherapy to intervention sites in mesothelioma: a systematic review and survey of UK practice.
Patients with malignant pleural mesothelioma (MPM), who undergo chest instrumentation, may develop seeding at the site of intervention, leading to subcutaneous tumour. This is believed to be reduced by the common practice of prophylactic irradiation to intervention tracts (PIT). However, evidence to support PIT is currently inadequate and contentious. We carried out a systematic search of published literature for articles relating to the incidence of chest wall intervention tract metastases and the use of PIT in mesothelioma. In addition, a survey of current practice was conducted in 54 UK oncology centres. Fourteen studies revealed an incidence of chest wall intervention tract metastases of 0-48% with a trend toward a higher rate of metastases for more invasive procedures. Three randomised controlled trials (RCTs), two prospective non-randomised studies and five retrospective series met the eligibility criteria to evaluate the role of PIT in MPM. Of the three RCTs, two did not support the use of PIT. None of the RCTs included patients who had received systemic chemotherapy. Of the oncology centres responding to the survey, 75% practiced PIT, and 80% would be interested in a trial to determine the efficacy of PIT. No consensus has been reached to support the use of PIT. However, most centres in the UK still offer PIT. There was widespread interest in a randomised controlled trial to establish PIT efficacy in the era of effective systemic chemotherapy for malignant pleural mesothelioma.


Evidence and Recommendations on the Use of Telemedicine for the Management of Arterial Hypertension: An International Expert Position Paper.
Telemedicine allows the remote exchange of medical data between patients and healthcare professionals. It is used to increase patients' access to care and provide effective healthcare services at a distance. During the recent coronavirus disease 2019 (COVID-19) pandemic, telemedicine has thrived and emerged worldwide as an indispensable resource to improve the management of isolated patients due to lockdown or shielding, including those with hypertension. The best proposed healthcare model for telemedicine in hypertension management should include remote monitoring and transmission of vital signs (notably blood pressure) and medication adherence plus education on lifestyle and risk factors, with video consultation as an option. The use of mixed automated feedback services with supervision of a multidisciplinary clinical team (physician, nurse, or pharmacist) is the ideal approach. The indications include screening for suspected hypertension, management of older adults, medically underserved people, high-risk hypertensive patients, patients with multiple diseases, and those isolated due to pandemics or national emergencies.


Crossing the smoking divide for young adults: expressions of stigma and identity among smokers and nonsmokers.
Denormalizing campaigns reframe smoking as an unappealing behavior, more likely to lead to social exclusion than inclusion. Social identity theory suggests this strategy will reinforce smoke-free norms and, as these become mainstream, decrease smoking prevalence. However, little is known about how these campaigns affect perceptions of smoking among young adult smokers and nonsmokers, or behavior toward smokers. A qualitative study was conducted to a) explore how smokers and smoking were perceived in an environment where smoking has become an increasingly unacceptable social behavior and b) examine whether and how this environment stigmatized smokers. About 14 group discussions and 4 in-depth interviews involving 86 participants, aged between 18 and 24 and of Māori, Pacific, and NZ European ethnicities, were conducted as part of a wider study examining young adults' responses to tobacco branding and plain packaging. The themes identified illustrated how nonsmokers' perception of smoking as illogical and self-destructive supported harsh reactions, including stigmatizing behaviors that antagonized smokers. Nonsmokers, who recognized smoking's addictiveness, were more empathic and less judgmental of smokers. Including empathic content in smoking denormalizing campaigns may reduce judgmental reactions that inadvertently create a gulf between status of young adult smokers and nonsmokers. A supportive/empathic tobacco-control denormalization approach could enhance young adult smokers' willingness to make the transition from smoker to smoke free and elicit stronger support for their efforts from nonsmokers.


Treatment of complicated skin and soft tissue infections.
Skin and soft tissue infections (SSTIs) may produce substantial morbidity and mortality rates, particularly those classified as complicated or necrotizing. To weigh the strength of recommendations using the Grades of Recommendation, Assessment, Development and Evaluation (GRADE) methodology and to provide evidence-based recommendations for diagnosis and management for SSTIs. Computerized identification of published research and review of relevant articles. All published reports on the management of complicated and necrotizing SSTIs were evaluated by an expert panel of members of the Surgical Infection Society according to published guidelines for evidence-based medicine. The quality of the evidence was judged by the GRADE methodology and criteria. Practice surveys, pharmacokinetic studies, and reviews or duplicative publications presenting primary data already considered were excluded from analysis. Information on demographics, study dates, microbiology findings, antibiotic type, surgical interventions, infection-related outcomes, and the methodologic quality of the studies was extracted. Results were submitted to the Therapeutic Agents Committee of the Surgical Infection Society for review prior to creation of the final consensus document. Current surgical and antibiotic management of complicated SSTIs is based on a small number of studies that often have insufficient power to draw well-supported conclusions, with the exception of antimicrobial therapy for non-necrotizing soft tissue infections, for which ample data are available.


Prediction of extreme ozone levels in Barcelona, Spain.
Barcelona is one of the most polluted cities in Western Europe, although our levels of air pollution are within the World Health Organisation air quality guidelines. However, high concentrations of air pollution have not been studied yet. Ground ozone levels is a topic of considerable environmental concern, since excessive level of ozone are taken as indicative of high pollution. In terms of the air quality guidelines ozone levels higher than 100 microg m(-3) can start to be health-hazards for human health. Our objective is to report a detailed analysis of ozone data exceeding the thresholds established by the air quality guidelines. Data analysed were collected in two measurement stations in Barcelona, for the reference period 1991--1996. Applying statistical techniques commonly used in the analysis of extreme values, mainly the Peak Over Threshold method was used for in this study. The analysis reveal that the ozone threshold values for the protection of human health has exceeded many times in both stations. The estimated return values for 3, 10, and 40 yr exceed the threshold value for information to the public of almost once in both stations, also it seems to be unlikely that the threshold value for warning to the public will be exceeded in 40 yr.


Electronic Tools to Bridge the Language Gap in Health Care for People Who Have Migrated: Systematic Review.
People who have migrated or with a language barrier may face significant hurdles in accessing health care. Some apps have been specifically developed to facilitate the dialogue between health care professionals and people who have migrated who have low-level language proficiency or to promote health among people who have migrated. We conducted a systematic review to investigate development, acceptability, and effectiveness of these types of apps. We conducted a search of PubMed, Scopus, and Embase databases. We included all study designs (qualitative, quantitative, mixed) reporting development, evaluation of efficacy, or acceptability of apps facilitating dialogue with a health professional or promoting health for people who have migrated, minorities, or tourists with a language barrier, using any outcome. Two researchers selected the studies independently. We collected general information about the app, information about health literacy and cultural adaptation, information about the development of the app, evidence on acceptability or efficacy, and information on app use. Data were collected by 2 researchers independently and results were reviewed to verify agreement and reported according to PRISMA (Preferred Reporting Items for Systematic Review and Meta-analysis). Positive results for translation apps included better communication, but with possible limitations, and reduced consultation time. Positive results for health promotion apps included improved quality of life and better management of chronic illnesses. Overall, the apps had good levels of acceptability, though only half had their efficacy evaluated. In those evaluations, the endpoints were mostly related to reported behavior change and knowledge improvement, which is common for evaluations of health promotion programs. In the future, as more health apps are created, it is essential that apps that claim to have a public health objective undergo a rigorous evaluation of their acceptability, efficacy, and actual use. Indicators of outcomes beyond changes in behavior and knowledge should be reported; change in health status or access to care should also be reported. This systematic review has helped us note the characteristics associated with improved acceptability and efficacy, which can be helpful for the development of future apps.


Arthritis imaging using a near-infrared fluorescence folate-targeted probe.
A recently developed near-infrared fluorescence-labeled folate probe (NIR2-folate) was tested for in vivo imaging of arthritis using a lipopolysaccharide intra-articular injection model and a KRN transgenic mice serum induction mouse model. In the lipopolysaccharide injection model, the fluorescence signal intensity of NIR2-folate (n = 12) and of free NIR2 (n = 5) was compared between lipopolysaccharide-treated and control joints. The fluorescence signal intensity of the NIR2-folate probe at the inflammatory joints was found to be significantly higher than the control normal joints (up to 2.3-fold, P < 0.001). The NIR2-free dye injection group showed a persistent lower enhancement ratio than the NIR2-folate probe injection group. Excessive folic acid was also given to demonstrate a competitive effect with the NIR2-folate. In the KRN serum transfer model (n = 4), NIR2-folate was applied at different time points after serum transfer, and the inflamed joints could be detected as early as 30 hours after arthritogenic antibody transfer (1.8-fold increase in signal intensity). Fluorescence microscopy, histology, and immunohistochemistry validated the optical imaging results. We conclude that in vivo arthritis detection was feasible using a folate-targeted near-infrared fluorescence probe. This receptor-targeted imaging method may facilitate improved arthritis diagnosis and early assessment of the disease progress by providing an in vivo characterization of active macrophage status in inflammatory joint diseases.


Cognitive disorders in type 2 diabetic patients with recognized depression.
The most important factors in the pathogenesis of cognitive disorders in diabetes mellitus (DM) are metabolic changes directly caused by hyperglycemia. Impairment of cognitive function is accompanied by a decrease in cerebral insulin. Insulin improves cognitive processes, including somatosensoric cognitive functions. Cognitive disorders are especially pronounced if diabetic patients suffer also from depression. The evaluation of cognitive functions, especially sensomotoric skills were obtained from three study groups (aged 37-52 years): 30 healthy subjects, 40 diabetic patients with clinically documented depression and 30 depressive patients without DM. The sensomotoric skills were carried out using a SPS-2001E apparatus - a computer working stress simulator. The examination with Beck Depression Inventory (BDI) were also performed in all the investigated persons. Sensomotoric skills investigation revealed slight cognitive disorders in the early stages DM (subgroup IA) and its intensification according to increasing BDI scores and HbA1c blood concentration (from subgroup IA to IC). Intensification of sensomotoric cognitive disorders was significantly higher (p<0.001) in the depressive patients with DM (group I), than in these without DM (group III). The obtained results suggest that coincidence of aggravating factors, such as depression and metabolic disorders in DM, may cause mutual interactions leading to premature and more intense cognitive impairment. The method proposed by the authors may serve as a screening examination in early diagnosis of CNS disorders reflected by somatosensoric cognitive disorders. The presented work indicates importance of sensomotoric skills investigation for the early diagnosis of the nervous system damage related to DM.


Quantitative determination of trinitroglycerin in human plasma.
We developed a simplified method for quantitative measurement of trinitroglycerin in human plasma using hexane extraction and analysis by gas-liquid chromatography with electron-capture detection. This assay was linear from 0.5-60 ng/ml. Sensitivity and reproducibility were +/- 0.5 ng/ml. We used this assay to evaluate the pharmacodynamics of trinitroglycerin in 14 patients. Maximum plasma levels were similar with trinitroglycerin given by constant intravenous infusion (1.6 +/- 0.4 ng/ml (SEM)), transcutaneously (2.3 +/- 0.6 ng/ml), or sublingually (1.6 +/- 0.6 ng/ml). Despite similar levels and hemodynamic responses after intravenous trinitroglycerin, the dose range was wide (37.5-175 microg/min, n = 5), emphasizing the need to individualize therapy. In normal volunteers on no other drugs, the plasma level time course followed changes in heart rate better than blood pressure changes. Use of the trinitroglycerin assay may enhance optimization of trinitroglycerin therapy when administered by different methods.


Exploring structural relationships between blood alcohol concentration and signs and clinical assessment of intoxication in alcohol-involved injury cases.
Although the relationship between the Y90 (blood alcohol concentration, BAC) and Y91 (clinician intoxication assessment) ICD-10 codes has received attention recently, the role of 10 signs of intoxication in the Y91-Y90 relationship has not been studied yet. This work examines these signs in the estimation of alcohol intoxication levels of patients in medical settings. Collected and analyzed were data on 1997 injured emergency room patients from 17 countries worldwide reporting drinking prior to injury or presenting with a non-zero BAC from 17 countries worldwide. A model is estimated describing how the 10 signs inform the Y91, Y90 prediction with the goal of the use of observations on patients in place of a biological measure. Signs were consistent with a single underlying construct that strongly predicted Y91. Smell of alcohol on breath predicted Y91 above its contribution through the construct and was stronger for those with tolerance to alcohol than for those without. Controlling for Y91, no sign further contributed to prediction of Y90 indicating that Y91 incorporated all intoxication sign information in predicting Y90. Variance explained was high for Y91 (R(2) = 0.84) and intoxication signs (above 0.72 for all but smell on the breath, 0.57) and lower for Y90 (0.38). Intoxication assessments are well predicted by overall intoxication severity, which itself is well represented by intoxication signs along with differential emphasis on smell of alcohol on breath, especially for those with alcohol tolerance. However, BAC levels remain largely unexplained by intoxication signs with a clinician's assessment serving as the primary predictive measure.


Branchial lesions associated with abundant apoptotic cells in oysters Ostrea edulis of Galicia (NW Spain).
An experiment to evaluate differences in growth, mortality and disease susceptibility among Ostrea edulis stocks was performed. Five families were produced from each of 4 oyster populations (Irish, Greek and 2 Galician). The spat were transferred to a raft in the Ria de Arousa (Galicia, Spain) for grow-out. Monthly samples of each family were histologically processed from 2001 to 2003. One of the pathological conditions discovered by this study was the occurrence of extensive branchial lesions characterized by haemocytic infiltration and loss of branchial architecture. Furthermore, abundant atypical cells occurred among the haemocytes in the lesions in the branchial connective and epithelial tissues, but rarely in the mantle. These cells were contracted in size with nuclei showing chromatin condensation and fragmentation. Some nuclear chromatin aggregated under the nuclear membranes into crescent shapes, whereas others were uniformly dense. Those characteristics suggested that the cells were apoptotic haemocytes, which was confirmed by transmission electron microscopy (TEM) and by a terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) assay using the Apoptag Kit on paraffin sections. A low prevalence of gill lesions was detected in some, but not all, families of every origin peaking in July 2002 and April 2003. No etiologic agent was identified by either histology or TEM; thus, the cause of the abundance of apoptotic cells remains unclear.


Infrared sauna in patients with rheumatoid arthritis and ankylosing spondylitis. A pilot study showing good tolerance, short-term improvement of pain and stiffness, and a trend towards long-term beneficial effects.
To study the effects of infrared (IR) Sauna, a form of total-body hyperthermia in patients with rheumatoid arthritis (RA) and ankylosing spondylitis (AS) patients were treated for a 4-week period with a series of eight IR treatments. Seventeen RA patients and 17 AS patients were studied. IR was well tolerated, and no adverse effects were reported, no exacerbation of disease. Pain and stiffness decreased clinically, and improvements were statistically significant (p < 0.05 and p < 0.001 in RA and AS patients, respectively) during an IR session. Fatigue also decreased. Both RA and AS patients felt comfortable on average during and especially after treatment. In the RA and AS patients, pain, stiffness, and fatigue also showed clinical improvements during the 4-week treatment period, but these did not reach statistical significance. No relevant changes in disease activity scores were found, indicating no exacerbation of disease activity. In conclusion, infrared treatment has statistically significant short-term beneficial effects and clinically relevant period effects during treatment in RA and AS patients without enhancing disease activity. IR has good tolerability and no adverse effects.


Who should be responsible for supporting individuals with mental health problems? A critical literature review.
Individuals with mental health problems have many support needs that are often inadequately met; however, perceptions of who should be responsible for meeting these needs have been largely unexplored. Varying perceptions may influence whether, how, and to what extent relevant stakeholders support individuals with mental health problems. To critically evaluate the literature to determine who different stakeholders believe should be responsible for supporting individuals with mental health problems, what factors shape these perceptions, and how they relate to one another. A critical literature review was undertaken. Following an extensive literature search, the conceptual contributions of relevant works were critically evaluated. A concept map was created to build a conceptual framework of the topic. Views of individual versus societal responsibility for need provision and health; the morality of caring; and attributions of responsibility for mental illness offered valuable understandings of the review questions. Creating a concept map revealed that various interrelated factors may influence perceptions of responsibility. Varying perceptions of who should be responsible for supporting individuals with mental health problems may contribute to unmet support needs among this group. Our critical review helps build a much-needed conceptual framework of factors influencing perceptions of responsibility. Such a framework is essential as these views iteratively shape and reflect the complex divisions of mental healthcare roles and responsibilities. Understanding these perceptions can help define relevant stakeholders' roles more clearly, which can improve mental health services and strengthen stakeholder accountability.


Socioeconomic inequalities in health. No easy solution.
Socioeconomic status (SES) is strongly associated with risk of disease and mortality. Universal health insurance is being debated as one remedy for such health inequalities. This article considers mechanisms through which SES affects health and argues that a broader and more comprehensive approach is needed. Published articles surveyed using MEDLINE and review articles and bibliographies. Research is reviewed on the association of SES with health outcomes in different countries, including those with universal health coverage. Socioeconomic status relates to health at all levels of the SES hierarchy, and access to care accounts for little of this association. Other mechanisms are suggested and implications for policy and clinical practice are discussed. Health insurance coverage alone is not likely to reduce significantly SES differences in health. Attention should be paid both in policy decisions and in clinical practice to other SES-related factors that may influence patterns of health and disease.


Durable Clinical Benefit With Nivolumab Plus Ipilimumab in DNA Mismatch Repair-Deficient/Microsatellite Instability-High Metastatic Colorectal Cancer.
Purpose Nivolumab provides clinical benefit (objective response rate [ORR], 31%; 95% CI, 20.8 to 42.9; disease control rate, 69%; 12-month overall survival [OS], 73%) in previously treated patients with DNA mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) metastatic colorectal cancer (mCRC); nivolumab plus ipilimumab may improve these outcomes. Efficacy and safety results for the nivolumab plus ipilimumab cohort of CheckMate-142, the largest single-study report of an immunotherapy combination in dMMR/MSI-H mCRC, are reported. Patients and Methods Patients received nivolumab 3 mg/kg plus ipilimumab 1 mg/kg once every 3 weeks (four doses) followed by nivolumab 3 mg/kg once every 2 weeks. Primary end point was investigator-assessed ORR. Results Of 119 patients, 76% had received ≥ two prior systemic therapies. At median follow-up of 13.4 months, investigator-assessed ORR was 55% (95% CI, 45.2 to 63.8), and disease control rate for ≥ 12 weeks was 80%. Median duration of response was not reached; most responses (94%) were ongoing at data cutoff. Progression-free survival rates were 76% (9 months) and 71% (12 months); respective OS rates were 87% and 85%. Statistically significant and clinically meaningful improvements were observed in patient-reported outcomes, including functioning, symptoms, and quality of life. Grade 3 to 4 treatment-related adverse events (AEs) occurred in 32% of patients and were manageable. Patients (13%) who discontinued treatment because of study drug-related AEs had an ORR (63%) consistent with that of the overall population. Conclusion Nivolumab plus ipilimumab demonstrated high response rates, encouraging progression-free survival and OS at 12 months, manageable safety, and meaningful improvements in key patient-reported outcomes. Indirect comparisons suggest combination therapy provides improved efficacy relative to anti-programmed death-1 monotherapy and has a favorable benefit-risk profile. Nivolumab plus ipilimumab provides a promising new treatment option for patients with dMMR/MSI-H mCRC.


The psychological characteristics and health related behavior of adolescents: the possible of social physique anxiety and gender.
The purpose of this study was to examine whether or not the social physique anxiety level and gender have an influence on psychological characteristics and health related behavior of adolescents. Five hundred and ninety eight female (M(age) = 14.95, SD = .70 years) and three hundred and eighty four male (M(age) = 15.08, SD = .76 years) adolescents voluntarily participated in this study. The Social Physique Anxiety Scale (SPAS), three subscales of the Physical Self-Description Questionnaire, and the Multidimensional Perfectionism Scale as indicators of psychological characteristics were administered to all participants. The Eating Attitude Test and Physical Activity Assessment Questionnaire were used to determine health related behavior. It was found that adolescents with high levels of SPA (HSPA) had more unfavourable eating attitudes, higher scores in socially-prescribed perfectionism, negative global physical self-worth and negative body related perceptions than those with low levels of SPA (LSPA). Physical activity levels of adolescents did not differ in the two SPA groups (high/low level). In addition, male adolescents in the present study were more physically active and had favorable eating attitudes and more positive self-perceptions of body fat and general physical self-worth than their female counterparts.


Influence of lipid A acylation pattern on membrane permeability and innate immune stimulation.
Lipid A, the hydrophobic anchor of lipopolysaccharide (LPS), is an essential component in the outer membrane of Gram-negative bacteria. It can stimulate the innate immune system via Toll-like receptor 4/myeloid differentiation factor 2 (TLR4/MD2), leading to the release of inflammatory cytokines. In this study, six Escherichia coli strains which can produce lipid A with different acylation patterns were constructed; the influence of lipid A acylation pattern on the membrane permeability and innate immune stimulation has been systematically investigated. The lipid A species were isolated and identified by matrix assisted laser ionization desorption-time of flight/tandem mass spectrometry. N-Phenyl naphthylamine uptake assay and antibiotic susceptibility test showed that membrane permeability of these strains were different. The lower the number of acyl chains in lipid A, the stronger the membrane permeability. LPS purified from these strains were used to stimulate human or mouse macrophage cells, and different levels of cytokines were induced. Compared with wild type hexa-acylated LPS, penta-acylated, tetra-acylated and tri-acylated LPS induced lower levels of cytokines. These results suggest that the lipid A acylation pattern influences both the bacterial membrane permeability and innate immune stimulation. The results would be useful for redesigning the bacterial membrane structure and for developing lipid A vaccine adjuvant.


Experimental requirement of the error-compensating five-frame interferogram-collecting sequence used in phase-shifting interferometry.
Digital phase-shifting interferometry (PSI), a technique widely used in optical testing, requires interferograms collected at optical phase differences separated by a definite phase step. The five-frame interferogram-collecting sequence suggested by Hariharan et al. [Appl. Opt. 26, 2504 (1987)] is extremely effective in significantly reducing the errors in height profiles derived using PSI that are caused by phase-step errors. In this Letter, we report on a class of five-frame sequence that, owing to its mathematical equivalence with the one suggested by Hariharan et al. and its ease of execution, is more commonly used but is much less effective in reducing the height profile errors caused by phase-step errors.


A design improvement in continuous blood sampling and analysis for glucose in rest and exercise.
In the past 15 yr attempts to sample blood continuously for glucose analysis with the AutoAnalyzer (Technicon Corp.) have been reasonably successful in resting subjects. However, they have sometimes required heparinization of the subjects so as to avoid clotting in the tubes of the system. To avoid the hazards of heparinization during exercise, a method is described for sampling and analyzing blood glucose continuously with the AutoAnalyzer, using a specially designed male Luer adaptor which fits into disposable Teflon catheters.


Protective roles of SIRT1 in atherosclerosis.
SIRT1 is a NAD (+) -dependent class III histone deacetylase (HDAC) that mediates the effects of caloric restriction on lifespan and metabolic pathways in various organisms. It deacetylates both histone and non-histone proteins, and targets proteins with diverse cellular and tissue functions. In the vasculature of rodent models SIRT1 mediates vasodilatation via eNOS-derived nitric oxide (NO) and scavenging reactive oxygen species (ROS). Recent studies demonstrated further protective roles of SIRT1 in vascular biology and atherosclerosis. In endothelial cells and macrophages SIRT1 has anti-inflammatory functions by downregulating the expression of various pro-inflammatory cytokines by interfering with the NF-kB signaling pathway. Deacetylation of RelA/p65-NF-kB by SIRT1 in macrophages also suppresses the expression of Lox-1, a scavenger receptor for oxidized low-density lipoproteins (oxLDL), thereby preventing macrophage foam cell formation. Moreover, SIRT1 has been shown to regulate the activity of Liver X-receptor (LXR), thereby promoting ABCA1-driven reverse cholesterol transport in plaque macrophages. Finally, SIRT1 suppresses the expression of endothelial tissue factor (coagulation factor III) and hence exerts anti-thrombotic properties. These findings indicate atheroprotective effects of SIRT1 in atherogenesis and highlight the need for translational research from bench-to-bedside. Indeed, SIRT1 activators are available for experimental research and undergo clinical testing. Taken together, these studies suggest SIRT1 activation as a promising therapeutic approach in atherosclerosis. Further studies are necessary to better understand the exact role of SIRT1 in the protagonist cells orchestrating atherogenesis and to identify the specificity, target effects and putative off-target effects of these promising SIRT1 activators.


A survey on awareness, knowledge, and attitudes toward epilepsy in an urban community in Turkey.
Epilepsy is one of the most common chronic neurological disorders with a high prevalence. Epileptic people and their family members suffer more from social stigma than the disorder itself. Among various complex reasons knowledge and awareness about epilepsy are the two important factors underlying discriminatory attitudes towards epileptic people. Community pharmacists play a major role in the care of these patients. In this study we mainly aimed to gain insights into the knowledge and awareness of and attitudes (AKA) towards epilepsy both in epileptic and healthy individuals in an urban community. To this end we also aimed at developing a reliable and valid measurement tool to assess AKA levels. This study was conducted in 13 community pharmacies with 219 respondents. Factor analysis yielded three clear subscales. It was found that a vast majority of the participants were familiar with epilepsy; yet only 18 of them had detailed information. The community pharmacists were indicated as a main source of information about epilepsy at the same rate to that of physicians. Although most of the respondents knew that epilepsy was not a form of mental illness only about one forth of them knew the real cause. More than half of the respondents supported the epileptics' socialization in the community. We believe that the questionnaire developed in the study is a promising instrument for determining educational needs and offering guidance to healthcare professionals in developing standardized educational tools and programs.


Sex differences of high-risk carotid atherosclerotic plaque with less than 50% stenosis in asymptomatic patients: an in vivo 3T MRI study.
Men have a greater prevalence of high-risk carotid plaque features associated with stroke compared with women who have ≥50% stenosis, but little is known about these features in less significantly stenotic carotid arteries. This study aims to evaluate sex differences in complicated carotid plaque features in asymptomatic patients with <50% stenosis. Ninety-six patients (50 men, 46 women) with <50% carotid stenosis on MRA who had been referred for analysis of contralateral >50% carotid stenosis were included. The associations between sex and plaque features as identified by 3T MR carotid plaque imaging were examined by using logistic and linear regression models controlling for demographic characteristics, MRA stenosis, and the presence of contralateral plaque features. The presence of a thin/ruptured fibrous cap (16% versus 2%, adjusted odds ratio = 8.57, P = .047), IPH (24% versus 6%, adjusted odds ratio = 4.53, P = .027), and American Heart Association type VI plaque (26% versus 6%, adjusted odds ratio = 5.04, P = .017) was significantly higher in men than in women. These associations remained significant following adjustment for contralateral plaque features. Men demonstrated a larger percentage volume of LR/NC (median, 1.66% versus -0.21%; P < .01). Calcification was not significantly associated with sex. There is a sex difference of higher risk carotid plaque features during the early stage of disease seen in patients recruited for MR imaging evaluation of contralateral moderate-to-severe stenosis. Given the potential of using LR/NC without or with IPH to monitor therapy, these results indicate the possible importance of sex-based management in patients with asymptomatic carotid atherosclerosis across all stages of carotid stenosis.