Ultrastructure of the anterior intestinal epithelia of the orange-spotted grouper Epinephelus coioides larvae under different feeding regimes.
Enterocytes of the anterior to midsection of the intestine in grouper Epinephelus coioides larvae were compared among different treatments: unfed to the point-of-no-return (PNR), fed natural food only, and co-fed natural food and artificial diet. On day 3, the nutritional condition of unfed grouper larvae regressed with its reduced enterocyte heights which were further degraded on day 4, the PNR, when all the enterocytes were in advanced stages of apoptosis. The apoptosis appeared to be internally directed via the mitochondria. Among day 3 fed larvae, enterocyte heights of those fed artificial diet did not differ from those fed natural food only. Dietary phospholipid deficiency was indicated in larvae co-fed artificial diet on day 3 with an unusually large chylomicron opening into the inter-enterocyte space, and on days 6 and 33 by intestinal steatosis. On day 19, scant to absent lipid droplets in enterocytes of larvae disclosed heightened nutritional requirement preparatory to metamorphosis. As observed in unfed day 3 and premetamorphic day 19 E. coioides, larvae undergoing critical periods and starvation during development employ apoptosis to dispose of degenerated enterocytes that are phagocytosed by adjacent healthy enterocytes without causing inflammatory distress. Upon metamorphosis, grouper larval gut develops better immunity fitness with eosinophilic granule cells observed in the intestinal epithelia of day 33 larvae. Future studies on grouper larval nutrition may consider the appropriate dietary phospholipid levels and larval competence to biosynthesize highly unsaturated fatty acid from linoleic acid vis-à-vis the use of plant ingredients in artificial diet formulations. In vivo challenge tests may validate appropriate dietary nutrient supplementation and lead to better feed formulation, matching the varying energetic demands and digestive capacities of developing E. coioides larvae.


The Use of Physical Restraint in Norwegian Adult Psychiatric Hospitals.
Background. The use of coercion within the psychiatric services is problematic and raises a range of ethical, legal, and clinical questions. "Physical restraint" is an emergency procedure used in psychiatric hospitals to control patients that pose an imminent physical danger. We wished to review the literature published in scientific peer-reviewed journals describing studies on the use of physical restraint in Norway, in order to identify the current state of knowledge and directions for future research. Design. The databases PubMed, PsycINFO, CINAHL, Web of Science, and Embase were searched for studies relating to physical restraint (including holding) in Norwegian psychiatric hospitals, supplemented with hand searches. Results. 28 studies were included. Most of the studies were on rates of restraint, but there were also some studies on perceptions of patients and staff, case studies, and ethnographic studies. There was only one intervention study. There are differences in use between wards and institutions, which in part may be explained by differences in patient populations. Staff appear to be less negative to the use of restraint than patients. Conclusions. The studies that were identified were primarily concerned with rates of use and with patients' and staff's perspectives. More interventional studies are needed to move the field forward.


A new approach to stability analysis and stabilization of discrete-time T-S fuzzy time-varying delay systems.
This paper investigates the problems of stability analysis and stabilization for a class of discrete-time Takagi-Sugeno fuzzy systems with time-varying state delay. Based on a novel fuzzy Lyapunov-Krasovskii functional, a delay partitioning method has been developed for the delay-dependent stability analysis of fuzzy time-varying state delay systems. As a result of the novel idea of delay partitioning, the proposed stability condition is much less conservative than most of the existing results. A delay-dependent stabilization approach based on a nonparallel distributed compensation scheme is given for the closed-loop fuzzy systems. The proposed stability and stabilization conditions are formulated in the form of linear matrix inequalities (LMIs), which can be solved readily by using existing LMI optimization techniques. Finally, two illustrative examples are provided to demonstrate the effectiveness of the techniques proposed in this paper.


Tie2 activation protects against prothrombotic endothelial dysfunction in COVID-19.
Endothelial dysfunction accompanies the microvascular thrombosis commonly observed in severe COVID-19. Constitutively, the endothelial surface is anticoagulant, a property maintained at least in part via signaling through the Tie2 receptor. During inflammation, the Tie2 antagonist angiopoietin-2 (Angpt-2) is released from endothelial cells and inhibits Tie2, promoting a prothrombotic phenotypic shift. We sought to assess whether severe COVID-19 is associated with procoagulant endothelial dysfunction and alterations in the Tie2/angiopoietin axis. Primary HUVECs treated with plasma from patients with severe COVID-19 upregulated the expression of thromboinflammatory genes, inhibited the expression of antithrombotic genes, and promoted coagulation on the endothelial surface. Pharmacologic activation of Tie2 with the small molecule AKB-9778 reversed the prothrombotic state induced by COVID-19 plasma in primary endothelial cells. Lung autopsies from patients with COVID-19 demonstrated a prothrombotic endothelial signature. Assessment of circulating endothelial markers in a cohort of 98 patients with mild, moderate, or severe COVID-19 revealed endothelial dysfunction indicative of a prothrombotic state. Angpt-2 concentrations rose with increasing disease severity, and the highest levels were associated with worse survival. These data highlight the disruption of Tie2/angiopoietin signaling and procoagulant changes in endothelial cells in severe COVID-19. Our findings provide rationale for current trials of Tie2-activating therapy with AKB-9778 in COVID-19.


Differential transmission of actin motion within focal adhesions.
Cell migration requires the transmission of motion generated in the actin cytoskeleton to the extracellular environment through a complex assembly of proteins in focal adhesions. We developed correlational fluorescent speckle microscopy to measure the coupling of focal-adhesion proteins to actin filaments. Different classes of focal-adhesion structural and regulatory molecules exhibited varying degrees of correlated motions with actin filaments, indicating hierarchical transmission of actin motion through focal adhesions. Interactions between vinculin, talin, and actin filaments appear to constitute a slippage interface between the cytoskeleton and integrins, generating a molecular clutch that is regulated during the morphodynamic transitions of cell migration.


Understanding North American yoga therapists' attitudes, skills and use of evidence-based practice: A cross-national survey.
Little is known about the adoption of evidence-based practice (EBP) by yoga therapists (YTs). To determine the attitudes, skills, training, use, barriers and facilitators to the use of EBP amongst North American YTs DESIGN: Cross-sectional, descriptive survey METHODS: Self-identified YTs practicing in North America were invited to participate in an online survey. YT attitudes, skills, training, utilisation, barriers to use, and facilitators of EBP use were measured using the 84-item Evidence-Based practice Attitude and utilization SurvEy (EBASE). 367 members responded (∼20% of eligible participants). Attitudes towards EBP were generally positive with 88% agreeing that professional literature and research findings were useful for the practice of yoga therapy. Most (80%) were interested in improving their skills and the majority agreed that EBP improves the quality of care (68%), assists in making decisions (74%) and takes into account the YTs clinical experience when making clinical decisions (59%). Moderate to moderately-high levels of perceived skill in EBP were reported mostly utilizing online search engines (51%). Lack of clinical evidence was the only notable barrier to uptake reported by YTs (48%). Facilitators to EBP included access to online EBP education materials (70.6%), ability to download full-text journal articles and access to free online databases in the workplace (67.3%). North American YTs report positive attitudes, moderate to moderately-high levels of perceived skill and moderate uptake of EBP. This aligns them with other complementary and integrative health practitioners. Initiatives to support the adoption of EBP are proposed as a means of improving best practice in yoga therapy.


The importance of cellular internalization of antibody-targeted carbon nanotubes in the photothermal ablation of breast cancer cells.
Single-walled carbon nanotubes (CNTs) convert absorbed near infrared (NIR) light into heat. The use of CNTs in the NIR-mediated photothermal ablation of tumor cells is attractive because the penetration of NIR light through normal tissues is optimal and the side effects are minimal. Targeted thermal ablation with minimal collateral damage can be achieved by using CNTs attached to tumor-specific monoclonal antibodies (MAbs). However, the role that the cellular internalization of CNTs plays in the subsequent sensitivity of the target cells to NIR-mediated photothermal ablation remains undefined. To address this issue, we used CNTs covalently coupled to an anti-Her2 or a control MAb and tested their ability to bind, internalize, and photothermally ablate Her2(+) but not Her2(-) breast cancer cell lines. Using flow cytometry, immunofluorescence, and confocal Raman microscopy, we observed the gradual time-dependent receptor-mediated endocytosis of anti-Her2-CNTs whereas a control MAb-CNT conjugate did not bind to the cells. Most importantly, the Her2(+) cells that internalized the MAb-CNTs were more sensitive to NIR-mediated photothermal damage than cells that could bind to, but not internalize the MAb-CNTs. These results suggest that both the targeting and internalization of MAb-CNTs might result in the most effective thermal ablation of tumor cells following their exposure to NIR light.


The dynamic range paradox: a central auditory model of intensity change detection.
In this paper we use empirical loudness modeling to explore a perceptual sub-category of the dynamic range problem of auditory neuroscience. Humans are able to reliably report perceived intensity (loudness), and discriminate fine intensity differences, over a very large dynamic range. It is usually assumed that loudness and intensity change detection operate upon the same neural signal, and that intensity change detection may be predicted from loudness data and vice versa. However, while loudness grows as intensity is increased, improvement in intensity discrimination performance does not follow the same trend and so dynamic range estimations of the underlying neural signal from loudness data contradict estimations based on intensity just-noticeable difference (JND) data. In order to account for this apparent paradox we draw on recent advances in auditory neuroscience. We test the hypothesis that a central model, featuring central adaptation to the mean loudness level and operating on the detection of maximum central-loudness rate of change, can account for the paradoxical data. We use numerical optimization to find adaptation parameters that fit data for continuous-pedestal intensity change detection over a wide dynamic range. The optimized model is tested on a selection of equivalent pseudo-continuous intensity change detection data. We also report a supplementary experiment which confirms the modeling assumption that the detection process may be modeled as rate-of-change. Data are obtained from a listening test (N = 10) using linearly ramped increment-decrement envelopes applied to pseudo-continuous noise with an overall level of 33 dB SPL. Increments with half-ramp durations between 5 and 50,000 ms are used. The intensity JND is shown to increase towards long duration ramps (p<10(-6)). From the modeling, the following central adaptation parameters are derived; central dynamic range of 0.215 sones, 95% central normalization, and a central loudness JND constant of 5.5×10(-5) sones per ms. Through our findings, we argue that loudness reflects peripheral neural coding, and the intensity JND reflects central neural coding.


Gastroesophageal sphincter: a model.
There is substantial experimental and anatomic evidence suggesting that the human lower esophageal sphincter is not a muscular ring but has its correlate in the arrangement of the so-called muscular clasps and oblique sling fibers at the gastroesophageal junction. We assessed the mode of action of these distinct muscle units in a mechanical model. The arrangement of the clasp and sling fibers at the gastroesophageal junction was simulated with two elastic bands placed perpendicularly around the gastroesophageal junction of four pig specimens. Rapid pullback manometry with four radially oriented pressure transducers was performed in each specimen. The opening pressure was determined, and three-dimensional pressure images were constructed based on the manometric readings. The elastic bands established a competent high-pressure zone at the level of the gastroesophageal junction. The three-dimensional pressure images matched those usually observed in vivo in normal human volunteers. The vector volume of the high-pressure zone correlated with the opening pressure while individual resting pressure values and length of the high-pressure zone were not sufficient to estimate the competence of the gastroesophageal junction in the model. This model supports the contention that the combined action of the clasp and sling fibers establishes the manometric lower esophageal sphincter in humans.


Considering unseen arrivals in predictions of establishment risk based on border biosecurity interceptions.
Assessing species establishment risk is an important task used for informing biosecurity activities aimed at preventing biological invasions. Propagule pressure is a major contributor to the probability of invading species establishment; however, direct assessment of numbers of individuals arriving is virtually never possible. Inspections conducted at borders by biosecurity officials record counts of species (or higher-level taxa) intercepted during inspections, which can be used as proxies for arrival rates. Such data may therefore be useful for predicting species establishments, though some species may establish despite never being intercepted. We present a stochastic process-based model of the arrival-interception-establishment process to predict species establishment risk from interception count data. The model can be used to estimate the probability of establishment, both for species that were intercepted and species that had no interceptions during a given observation period. We fit the stochastic model to data on two insect families, Cerambycidae and Aphididae, that were intercepted and/or established in the United States or New Zealand. We also explore the effects of variation in model parameters and the inclusion of an Allee effect in the establishment probability. Although interception data sets contain much noise due to variation in inspection policy, interception effort and among-species differences in detectability, our study shows that it is possible to use such data for predicting establishments and distinguishing differences in establishment risk profile between taxonomic groups. Our model provides a method for predicting the number of species that have breached border biosecurity, including both species detected during inspections but also "unseen arrivals" that have never been intercepted, but have not yet established a viable population. These estimates could inform prioritization of different taxonomic groups, pathways or identification effort in biosecurity programs.


Combined resveratrol, quercetin, and catechin treatment reduces breast tumor growth in a nude mouse model.
Grape polyphenols can act as antioxidants, antiangiogenics, and selective estrogen receptor (ER) modifiers and are therefore especially relevant for gynecological cancers such as breast cancer. The major polyphenols of red wine (resveratrol, quercetin, and catechin) have been individually shown to have anticancer properties. However, their combinatorial effect on metastatic breast cancers has not been investigated in vivo. We tested the effect of low dietary concentrations of resveratrol, quercetin, and catechin on breast cancer progression in vitro by analyzing cell proliferation and cell cycle progression. The effects of these compounds on fluorescently tagged breast tumor growth in nude mice were assessed using in situ fluorescence image analysis. Individual polyphenols at 0.5 microM neither decreased breast cancer cell proliferation nor affected cell cycle progression in vitro. However, a combination of resveratrol, quercetin, and catechin at 0.5, 5, or 20 microM each significantly reduced cell proliferation and blocked cell cycle progression in vitro. Furthermore, using in situ image analysis, we determined that combined dietary polyphenols at 0.5, 5, or 25 mg/kg reduced primary tumor growth of breast cancer xenografts in a nude mouse model. Peak inhibition was observed at 5 mg/kg. These results indicate that grape polyphenols may inhibit breast cancer progression.


Global prevalence and phylogeny of hepatitis B virus (HBV) drug and vaccine resistance mutations.
Vaccination and anti-viral therapy with nucleos(t)ide analogues (NAs) are key approaches to reducing the morbidity, mortality and transmission of hepatitis B virus (HBV) infection. However, the efficacy of these interventions may be reduced by the emergence of drug resistance-associated mutations (RAMs) and/or vaccine escape mutations (VEMs). We have assimilated data on the global prevalence and distribution of HBV RAMs/VEMs from publicly available data and explored the evolution of these mutations. We analysed sequences downloaded from the HBV Database and calculated prevalence of 41 RAMs and 38 VEMs catalogued from published studies. We generated maximum likelihood phylogenetic trees and used treeBreaker to investigate the distribution and estimated the age of selected mutations across tree branches. RAM M204I/V had the highest prevalence, occurring in 3.8% (109/2838) of all HBV sequences in our data set, and a significantly higher rate in genotype C at 5.4% (60/1102, p = 0.0007). VEMs had an overall prevalence of 1.3% (37/2837) and had the highest prevalence in genotype C and in Asia at 2.2% (24/1102; p = 0.002) and 1.6% (34/2109; p = 0.009), respectively. Phylogenetic analysis suggested that RAM/VEMs can arise independently of treatment/vaccine exposure. In conclusion, HBV RAMs/VEMs have been found globally and across genotypes, with the highest prevalence observed in genotype C. Screening for genotype and for resistance-associated mutations may help to improve stratified patient treatment. As NAs and HBV vaccines are increasingly being deployed for HBV prevention and treatment, monitoring for resistance and advocating for better treatment regimens for HBV remains essential.


Anderseniella baltica gen. nov., sp. nov., a novel marine bacterium of the Alphaproteobacteria isolated from sediment in the central Baltic Sea.
A bacterial isolate from the Baltic Sea, designated strain BA141(T), was characterized for its physiological and biochemical features, fatty acid profile, pigment spectrum, DNA G+C content and phylogenetic position based on comparative 16S rRNA gene sequence analysis. The strain was isolated from the surface of sediment in a deep basin of the central Baltic Sea. Phylogenetic analysis of the 16S rRNA gene sequence revealed a clear affiliation with the Alphaproteobacteria, and showed that the closest phylogenetic relationship was with the genus Rhodobium. The G+C content of the DNA was 61.2 mol%. Cells of strain BA141(T) were red-pigmented, Gram-negative, pleomorphic to rod-shaped, non-motile and catalase- and oxidase-positive. Growth was observed at salinities of 0.8-6 %, with optimum growth at 1.5-3 %. The temperature range for growth was 10-37 degrees C, with optimum growth at 25-30 degrees C. The fatty acids were dominated by unsaturated fatty acids (>86 %); the number of fatty acids detected was very low, with 18 : 1omega7c (73 %) as the predominant fatty acid; other major fatty acids were 19 : 0 cyclo 8c (10 %) and 16 : 0 (8 %). The pigment spectrum indicated the presence of carotenoids and unknown pigment(s) with an absorption maximum at 430 nm, but not bacteriochlorophyll a. According to phylogenetic analysis based on the 16S rRNA gene sequence and the phenotypic features, strain BA141(T) represents a new genus and species. The name Anderseniella baltica gen. nov., sp. nov. is proposed, with strain BA141(T) (=CIP 109499(T)=LMG 24028(T)) as the type strain.


Advances in stem cell therapy for the treatment of Peyronie's disease.
Peyronie's disease (PD) is a connective tissue disorder of the penis characterized by fibrosis and plaque formation within the tunica albuginea. PD is characterized by painful penile curvature that impairs sexual intercourse. Stem cell therapy is one of the recent non-invasive treatment options for patients with PD and it has promising results. Stem cells are undifferentiated cells that are capable of self-renewal and differentiation, promoting the repair of tissues via their immunomodulatory and anti-inflammatory action. Adipose-derived stem cells (ADSC) are used most widely due to their abundant tissue source and ease of isolation. Multiple studies have indicated the efficacy of stem cell therapy as a potential treatment for fibrotic diseases. Clearly, ADSCs may represent a way to treat and prevent PD in both rat and human models. Further clinical studies are needed to confirm the efficacy of stem cell therapy for PD in humans.


Moss Kinesin-14 KCBP Accelerates Chromatid Motility in Anaphase.
KCBP is a microtubule (MT) minus-end-directed kinesin widely conserved in plants. It was shown in Arabidopsis that KCBP controls trichome cell shape by orchestrating MT and actin cytoskeletons using its tail and motor domains. In contrast, the KCBP knockout (KO) line in the moss Physcomitrella patens showed a defect in nuclear and organelle positioning in apical stem cells. Moss KCBP is postulated to transport the nucleus and chloroplast via direct binding to their membranes, since it binds to and transports liposomes composed of phospholipids in vitro. However, domains required for cargo transport in vivo have not been mapped. Here, we performed a structure-function analysis of moss KCBP. We found that the FERM domain in the tail region, which is known to bind to lipids as well as other proteins, is essential for both nuclear and chloroplast positioning, whereas the proximal MyTH4 domain plays a supporting role in chloroplast transport. After anaphase but prior to nuclear envelope re-formation, KCBP accumulates on the chromosomes, in particular at the centromeric region in a FERM-dependent manner. In the KCBP KO line, the rate of poleward chromosome movement in anaphase was reduced and lagging chromosomes occasionally appeared. These results suggest that KCBP binds to non-membranous naked chromosomes via an unidentified protein(s) for their transport. Finally, the liverwort orthologue of KCBP rescued the chromosome/chloroplast mis-positioning of the moss KCBP KO line, suggesting that the cargo transport function is conserved at least in bryophytes.Key words: kinesin, mitosis, chromosome segregation, kinetochore, dynein.


Impact of agriculture on the selection of insecticide resistance in the malaria vector Anopheles gambiae: a multigenerational study in controlled conditions.
Resistance of mosquitoes to insecticides is mainly attributed to their adaptation to vector control interventions. Although pesticides used in agriculture have been frequently mentioned as an additional force driving the selection of resistance, only a few studies were dedicated to validate this hypothesis and characterise the underlying mechanisms. While insecticide resistance is rising dramatically in Africa, deciphering how agriculture affects resistance is crucial for improving resistance management strategies. In this context, the multigenerational effect of agricultural pollutants on the selection of insecticide resistance was examined in Anopheles gambiae. An urban Tanzanian An. gambiae population displaying a low resistance level was used as a parental strain for a selection experiment across 20 generations. At each generation larvae were selected with a mixture containing pesticides and herbicides classically used in agriculture in Africa. The resistance levels of adults to deltamethrin, DDT and bendiocarb were compared between the selected and non-selected strains across the selection process together with the frequency of kdr mutations. A microarray approach was used for pinpointing transcription level variations selected by the agricultural pesticide mixture at the adult stage. A gradual increase of adult resistance to all insecticides was observed across the selection process. The frequency of the L1014S kdr mutation rose from 1.6% to 12.5% after 20 generations of selection. Microarray analysis identified 90 transcripts over-transcribed in the selected strain as compared to the parental and the non-selected strains. Genes encoding cuticle proteins, detoxification enzymes, proteins linked to neurotransmitter activity and transcription regulators were mainly affected. RT-qPCR transcription profiling of candidate genes across multiple generations supported their link with insecticide resistance. This study confirms the potency of agriculture in selecting for insecticide resistance in malaria vectors. We demonstrated that the recurrent exposure of larvae to agricultural pollutants can select for resistance mechanisms to vector control insecticides at the adult stage. Our data suggest that in addition to selected target-site resistance mutations, agricultural pollutants may also favor cuticle, metabolic and synaptic transmission-based resistance mechanisms. These results emphasize the need for integrated resistance management strategies taking into account agriculture activities.


Reduced risk of pneumonia after changes in anesthetic procedures for dogs receiving repeated anesthesia for radiation treatment.
Radiation therapy requires repeated anesthetic administration to patients who often have multiple comorbidities contributing to an increased rate of anesthetic complications such as pneumonia. This is a retrospective observational study in which data were collected from 146 medical records of dogs receiving repeat anesthesia for radiation treatment from prior to management changes and compared to data from 149 cases treated after completion of management changes. The objective was to determine if changes in case management protocol that were put in place decreased the risk of pneumonia development among these patients. Management changes that were made included the following: decrease in anticholinergic and pure-mu opioid use, change in positioning during intubation and recovery, prophylactic treatment of nausea, timing of cuff inflation and deflation, and aseptic handling of intubation equipment. There was a significant association between diagnosis of pneumonia and the following: pre- vs. post-changes to protocol, presence of a neurologic tumor, presence of respiratory disease, presence of megaesophagus, and number of radiation fractions completed. Diagnosis of pneumonia did not vary significantly by age group, body weight category, or sex. In a multivariable logistic regression model that controlled for the effects of the three concurrent diseases and fractions completed, the odds of being diagnosed with pneumonia were approximately 10 times greater among dogs anesthetized prior to management changes (odds ratio = 9.9, 95% CI = 2.0-48.7, P = 0.005).


CTL responses to a DNA vaccine encoding E7 gene of human papillomavirus type 16 from an Iranian isolate.
Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among female population worldwide. Specific human papillomaviruses and, most notably, HPV types 16 and 18 are recognized as being causally associated with cervical carcinomas. The early HPV type 16 genes, E6 and E7, directly participate in the in vitro transformation of primary human keratinocytes and represent an excellent target for immune therapy of HPV related disease. The aim of this study was the evaluation of the efficacy of a DNA vaccine containing human papillomaviruse type 16 E7 gene (Iranian isolate) in induction of CTL responses in an animal model. In this study, the expression vector containing HPV type 16 E7 gene was constructed and chosen as a model antigen in the development of a therapeutic DNA vaccine in an animal model. CTL responses, cytokine assay, lymphocyte stimulation test, CD4 and CD8 staining and flowcytometry were done for evaluating of the immune responses. Our findings indicate that the target DNA vaccine can induce an E7-specific CTL response, which is important in the lysis of infected tumor cells, compared to negative control (p<0.005) after in vivo immunization in the mouse system. The developed vaccine may be promising as an anti-cancer vaccine.


Genomic erosion and extensive horizontal gene transfer in gut-associated Acetobacteraceae.
Symbiotic relationships between animals and bacteria have profound impacts on the evolutionary trajectories of each partner. Animals and gut bacteria engage in a variety of relationships, occasionally persisting over evolutionary timescales. Ants are a diverse group of animals that engage in many types of associations with taxonomically distinct groups of bacterial associates. Here, we bring into culture and characterize two closely-related strains of gut associated Acetobacteraceae (AAB) of the red carpenter ant, Camponotus chromaiodes. Genome sequencing, assembly, and annotation of both strains delineate stark patterns of genomic erosion and sequence divergence in gut associated AAB. We found widespread horizontal gene transfer (HGT) in these bacterial associates and report elevated gene acquisition associated with energy production and conversion, amino acid and coenzyme transport and metabolism, defense mechanisms, and lysine export. Both strains have acquired the complete NADH-quinone oxidoreductase complex, plausibly from an Enterobacteriaceae origin, likely facilitating energy production under diverse conditions. Conservation of several lysine biosynthetic and salvage pathways and accumulation of lysine export genes via HGT implicate L-lysine supplementation by both strains as a potential functional benefit for the host. These trends are contrasted by genome-wide erosion of several amino acid biosynthetic pathways and pathways in central metabolism. We perform phylogenomic analyses on both strains as well as several free living and host associated AAB. Based on their monophyly and deep divergence from other AAB, these C. chromaiodes gut associates may represent a novel genus. Together, our results demonstrate how extensive horizontal transfer between gut associates along with genome-wide deletions leads to mosaic metabolic pathways. More broadly, these patterns demonstrate that HGT and genomic erosion shape metabolic capabilities of persistent gut associates and influence their genomic evolution. Using comparative genomics, our study reveals substantial changes in genomic content in persistent associates of the insect gastrointestinal tract and provides evidence for the evolutionary pressures inherent to this environment. We describe patterns of genomic erosion and horizontal acquisition that result in mosaic metabolic pathways. Accordingly, the phylogenetic position of both strains of these associates form a divergent, monophyletic clade sister to gut associates of honey bees and more distantly to Gluconobacter.


Viral and metabolic controls on high rates of microbial sulfur and carbon cycling in wetland ecosystems.
Microorganisms drive high rates of methanogenesis and carbon mineralization in wetland ecosystems. These signals are especially pronounced in the Prairie Pothole Region of North America, the tenth largest wetland ecosystem in the world. Sulfate reduction rates up to 22 μmol cm-3 day-1 have been measured in these wetland sediments, as well as methane fluxes up to 160 mg m-2 h-1-some of the highest emissions ever measured in North American wetlands. While pore waters from PPR wetlands are characterized by high concentrations of sulfur species and dissolved organic carbon, the constraints on microbial activity are poorly understood. Here, we utilized metagenomics to investigate candidate sulfate reducers and methanogens in this ecosystem and identify metabolic and viral controls on microbial activity. We recovered 162 dsrA and 206 dsrD sequences from 18 sediment metagenomes and reconstructed 24 candidate sulfate reducer genomes assigned to seven phyla. These genomes encoded the potential for utilizing a wide variety of electron donors, such as methanol and other alcohols, methylamines, and glycine betaine. We also identified 37 mcrA sequences spanning five orders and recovered two putative methanogen genomes representing the most abundant taxa-Methanosaeta and Methanoregulaceae. However, given the abundance of Methanofollis-affiliated mcrA sequences, the detection of F420-dependent alcohol dehydrogenases, and millimolar concentrations of ethanol and 2-propanol in sediment pore fluids, we hypothesize that these alcohols may drive a significant fraction of methanogenesis in this ecosystem. Finally, extensive viral novelty was detected, with approximately 80% of viral populations being unclassified at any known taxonomic levels and absent from publicly available databases. Many of these viral populations were predicted to target dominant sulfate reducers and methanogens. Our results indicate that diversity is likely key to extremely high rates of methanogenesis and sulfate reduction observed in these wetlands. The inferred genomic diversity and metabolic versatility could result from dynamic environmental conditions, viral infections, and niche differentiation in the heterogeneous sediment matrix. These processes likely play an important role in modulating carbon and sulfur cycling in this ecosystem.


A Vinyl Ether-Functional Polycarbonate as a Template for Multiple Postpolymerization Modifications.
A highly-reactive vinyl ether-functionalized aliphatic polycarbonate and its block copolymer were developed as templates for multiple post-polymerization conjugation chemistries. The vinyl ether-functional six-membered cyclic carbonate monomer was synthesized by a well-established two-step procedure starting from 2,2-bis(hydroxymethyl)propionic acid. An organobase-catalyzed ring-opening polymerization of the synthesized monomer afforded polycarbonates with pendant vinyl ether functionalities (PMVEC). The vinyl ether moieties on the resulting polymers were readily conjugated with hydroxyl- or thiol-containing compounds via three different post-polymerization modification chemistries - acetalization, thio-acetalization, and thiol-ene reaction. Acetal-functionalized polycarbonates were studied in depth to exploit their acid-labile acetal functionalities. Acetalization of the amphiphilic diblock copolymer of poly(ethylene glycol) methyl ether (mPEG) and PMVEC, mPEG113-b-PMVEC13, with the model hydroxyl compound 4- methylbenzyl alcohol resulted in a maximum of 42% acetal and 58% hydroxyl side chain groups. Nonetheless, the amphiphilicity of the block polymer allowed for its self-assembly in water to afford nanostructures, as characterized via dynamic light scattering and transmission electron microscopy. The kinetics of acetal cleavage within the block polymer micelles were examined in acidic buffered solutions (pH 4 and 5). In addition, mPEG-b-PMVEC and its hydrolyzed polymer mPEG-b-PMHEC (i.e., after full cleavage of acetals) exhibited minimal cytotoxicity to RAW 264.7 mouse macrophages, indicating that this polymer system represents a biologically non-hazardous material with pH-responsive activity.


Socioeconomic Differences and the Potential Role of Tribes in Young People's Food and Drink Purchasing Outside School at Lunchtime.
Socioeconomic deprivation has been linked to food consumption practices, but studies investigating the food environment around schools provide mixed findings. Peer influence and marketing cues are considered important influencers of young people's behaviors. This study used a tribal theory lens to investigate the factors affecting pupils' purchasing and consumption of food/drinks outside schools at lunchtime. A survey was conducted with 243 pupils from seven UK secondary schools of differing socioeconomic status (SES). A purchasing recall questionnaire (PRQ) was developed and administered online at the participating schools to capture food and drink purchasing, intake, and expenditure. No significant differences were found in terms of energy and nutrients consumed or food/drink expenditure between pupils from schools of lower and higher SES. Enjoyment of food shopping with friends was linked with higher food energy intake and spend. Higher susceptibility to peer influence was associated with greater influence from food advertising and endorsements. Without ignoring the impact that SES can have on young people's food choices, we suggest that tribal theory can be additionally used to understand pupils' eating behaviors and we present implications for social marketers and policy makers.


Protections against toxicity in the brains of rat with chronic fluorosis and primary neurons exposed to fluoride by resveratrol involves nicotinic acetylcholine receptors.
Protection of Resveratrol (RSV) against the neurotoxicity induced by high level of fluoride was investigated. Sprague-Dawley (SD) rats and their offspring, as well as cultures of primary neurons were divided randomly into four groups: untreated (control); treated with 50 mg RSV/kg/ (once daily by gavage) or (20 M in the cultured medium); exposed to 50 ppm F- in drinking water or 4 mmol/l in the cultured medium; and exposed to fluoride then RSV as above. The adult rats were treated for 7 months and the offspring sacrificed at 28 days of age; the cultured neurons for 48 h. For general characterization, dental fluorosis was assessed and the fluoride content of the urine measured (by fluoride-electrode) in the rates and the survival of cultured neurons monitored with the CCK-8 test. The spatial learning and memory of rats were assessed with the Morris water maze test. The levels of α7 and α4 nicotinic acetylcholine receptors (nAChRs) were quantified by Western blotting; and the activities of superoxide dismutase (SOD) and catalase (CAT), and the levels of malondialdehyde (MDA) and H2O2 assayed biochemically. The results showed that chronic fluorosis resulted in the impaired learning and memory in rats and their offspring, and more oxidative stress in both rat brains and cultured neurons, which may be associated the lower levels of α7 and α4 nAChR subunits. Interestingly, RSV attenuated all of these toxic effects by fluorosis, indicating that protection against the neurotoxicity of fluoride by RSV might be in mechanism involved enhancing the expressions of these nAChRs.


The role of the hepatitis C virus glycoproteins in infection.
HCV encodes two glycoproteins, E1 and E2, that are believed to be exposed on the surface of virions. These molecules are likely to be involved in viral interactions with the host immune response and responsible for mediating viral entry into target cells. They are obvious major components for prototype vaccine studies. Recently, E2 has been reported to bind to the tetraspan molecule CD81, which represents a putative receptor for HCV. Here, we discuss the role the HCV gps may play during infection, the contribution of E2 gp variation to HCV evasion from the immune response and possible implications of the E2-CD81 interaction for HCV pathogenesis.


Human megakaryocytes. III. Characterization in myeloproliferative disorders.
Abnormal proliferation of the megakaryocytic line was observed in the marrow tissue from patients with myeloproliferative disorders. Megakaryocytes were identified by immunofluorescence using distinct platelet protein markers. Plasma factor VIII antigen (factor VIII:AGN) and platelet glycoproteins IIb and IIIa were detected in normal mature and early megakaryocytes, as well as in a morphologically heterogeneous population of low density marrow cells regarded as atypical megakaryocytes. Atypical megakaryocytes were defined as oval/round 14-35-micron diameter blast-like mononuclear/multinucleated cells bearing platelet protein markers with distinct morphological features, including cytoplasmic vacuolation, variable nuclear/cytoplasmic ratios, and variable cytoplasmic granulation. Atypical megakaryocytes were observed in most chronic myelogenous leukemia (CML) patients and in two patients with polycythemia vera, representing between 60 and 1,840 cells/10(4) cells (less than 1.050 g Percoll/cu cm). No atypical megakaryocytes were found in (a) 20 normal controls, (b) two patients with essential thrombocythemia, (c) a patient with thrombocytosis secondary to acute bleeding, and (d) in two patients with CML. Atypical megakaryocytes appear to represent a single-cell population, as demonstrated by a series of double immunofluorescence assays using combinations of five different antiplatelet protein sera. There was a statistically significant correlation between the frequency of atypical megakaryocytes and the presence of immature forms of myeloid cells in blood. Analyses of Fc IgG receptors conducted with two different immunofluorescence systems have demonstrated that phenotypic similarities existed between atypical megakaryocytes and myeloproliferative platelet proteins and differentiation markers on megakaryocytes are useful in elucidating the pathophysiologic alterations occurring in the megakaryocytic compartment in patients with myeloproliferative disorders.


Effect of spinal construct stiffness on early fusion mass incorporation. Experimental study.
The relationship between initial spinal construct stiffness and the stiffness of the resulting fusion mass was studied by performing standardized 10-segment posterior spinal fusions in goats. Animals were divided into 5 groups based on type of spinal construct, using rods of different diameters (3.2 mm, 4.8 mm, 6.4 mm) with or without rigid crosslinking to produce constructs of different stiffnesses. Stiffness data on 28 animals were obtained by removing the spines en bloc, at 6 or 12 weeks postoperatively, and performing load-deformation testing in axial and torsional loading to determine the stiffness of the fusion masses (rods removed). The initial construct stiffnesses were also compared by ex vivo testing on spine specimens to correlate initial construct stiffness with eventual fusion mass stiffness. In axial testing, results showed stiffer fusion masses from larger diameter rod constructs compared with smaller rod constructs. This was similar to results of control testing on spine specimens ex vivo. Rigid crosslinking did not produce stiffer fusions in axial testing, due to a technical limitation of the button-wire implants used to segmentally fix the rods at each vertebra. In torsional testing, stiffer fusion masses resulted from using larger rods, and rigid Crosslinking also produced the stiffest fusion masses, which was consistent with ex vivo testing. In general, larger diameter (stiffer) rods produced stiffer fusion masses, and no evidence of stress shielding was found.


New PPARG mutation leads to lipodystrophy and loss of protein function that is partially restored by a synthetic ligand.
Familial partial lipodystrophy caused by mutations in the PPARG gene is characterised by altered distribution of subcutaneous fat, muscular hypertrophy and symptoms of metabolic syndrome. PPARG encodes peroxisome proliferator-activated receptor (PPAR)gamma, a nuclear hormone receptor playing a crucial role in lipid and glucose metabolism and in several other cellular regulatory processes. PPARG was screened for mutations by direct sequencing in two patients with lipodystrophy, one unaffected family member and 124 controls. Body composition was examined in affected patients, and they were investigated for abnormalities in laboratory results. Functional analysis of the mutant protein was assessed by determining transcriptional activity and possible interference with the wild-type protein. In two patients with familial partial lipodystrophy, we identified a nucleotide substitution in the PPARG gene. This mutation results in the substitution of aspartate by asparagine at residue 424 (D424N) in the ligand-binding domain of PPARgamma. The unaffected family member and all 124 controls did not carry this mutation. D424N PPARgamma had a significantly lower ability than wild-type PPARgamma to activate a PPARgamma-stimulated reporter gene, but did not exert a negative effect on the wild-type protein. Partial activation of D424N PPARgamma was achieved in the presence of the agonist rosiglitazone. We report a new PPARG mutation, D424N, which is located in the ligand-binding domain of the protein and leads to familial partial lipodystrophy. D424N PPARgamma exhibited a loss of function, which was partially restored by adding the PPARgamma agonist rosiglitazone, suggesting possible treatment potential of this agent.


Application of disaccharides alone and in combination, for the improvement of stability and particle properties of spray-freeze dried IgG.
Spray-freeze drying (SFD) is a recently applied method to develop pharmaceutical powders. This study aimed to analyze the competence of Trehalose, Mannitol, Lactose, and Sorbitol instability and aerosolization of Immunoglobulin G (IgG) via SFD. Induced soluble aggregates were quantified at 0 and 3 months, and 45 °C using size-exclusion chromatography. Conformation and thermogravimetric assessments were done by Fourier transform infrared spectroscopy and differential scanning calorimetry. Laser light scattering was performed to determine the particle sizes. Aerodynamic features were characterized by twin stage impinger and scanning electron microscopy. Although sugars/polyols preferably stabilized IgG following the process, storage stabilization was achieved in Trehalose, Trehalose-Lactose, Lactose, and Trehalose-Mannitol-based powders with soluble aggregates <5%. The conformation of antibody was preserved with β sheet content from 66.28% to 76.37%. Particle sizes ranged from 5.23 to 8.12 µm. Mannitol exhibited the best aerodynamic behavior, fine particle fraction (FPF: 70%) but high degree of protein aggregation during storage. SFD could favorably stabilize antibody using Trehalose and its combination with Lactose and Mannitol, and also, Lactose alone. Sorbitol disturbed IgG powder recovery. Incorporation of other types of excipient is required for efficient respiratory delivery of IgG molecules.


Multiple effects of physical activity on molecular and cognitive signs of brain aging: can exercise slow neurodegeneration and delay Alzheimer's disease?
Western countries are experiencing aging populations and increased longevity; thus, the incidence of dementia and Alzheimer's disease (AD) in these countries is projected to soar. In the absence of a therapeutic drug, non-pharmacological preventative approaches are being investigated. One of these approaches is regular participation in physical activity or exercise. This paper reviews studies that have explored the relationship between physical activity and cognitive function, cognitive decline, AD/dementia risk and AD-associated biomarkers and processes. There is now strong evidence that links regular physical activity or exercise to higher cognitive function, decreased cognitive decline and reduced risk of AD or dementia. Nevertheless, these associations require further investigation, more specifically with interventional studies that include long follow-up periods. In particular, relatively little is known about the underlying mechanism(s) of the associations between physical activity and AD neuropathology; clearly this is an area in need of further research, particularly in human populations. Although benefits of physical activity or exercise are clearly recognised, there is a need to clarify how much physical activity provides the greatest benefit and also whether people of different genotypes require tailored exercise regimes.


Health impact assessment for the sustainable futures of Salford.
The Salford Health Investment for Tomorrow and the Local Improvement Finance Trust proposals for the comprehensive redevelopment of Salford Royal Hospital and the provision of four integrated primary health and social centres in the City of Salford are a response to the challenge of the NHS Plan to modernize delivery and service provision. The scale of the proposed developments will have a major effect on the quality of life in Salford. This health impact assessment was undertaken to determine how these proposals could be amended to ensure that the impacts are optimised so that potential benefits to Salford's communities are realised to maximise their health and wellbeing. This involved a comprehensive prospective health impact assessment and a series of interviews with key stakeholders, community organisations, and local people to identify and prioritize potential positive and negative impacts of the proposals. The assessment pointed to a wide range of positive and negative impacts that provided the basis of a set of recommendations, which were linked directly to the themes of the Salford Community Plan. These related to setting employment, education, and training opportunities to maximise health and wellbeing and improve quality of life for all communities in Salford within the framework of the plan. In presenting the findings of the health impact assessment and discussing the recommendations, the paper presents a unique approach in health impact assessment reporting by offering perspectives from the assessor and the project's steering group.


Chemical warfare between fungus-growing ants and their pathogens.
Fungus-growing attine ants are under constant threat from fungal pathogens such as the specialized mycoparasite Escovopsis, which uses combined physical and chemical attack strategies to prey on the fungal gardens of the ants. In defence, some species assemble protective microbiomes on their exoskeletons that contain antimicrobial-producing Actinobacteria. Underlying this network of mutualistic and antagonistic interactions are an array of chemical signals. Escovopsis weberi produces the shearinine terpene-indole alkaloids, which affect ant behaviour, diketopiperazines to combat defensive bacteria, and other small molecules that inhibit the fungal cultivar. Pseudonocardia and Streptomyces mutualist bacteria produce depsipeptide and polyene macrolide antifungals active against Escovopsis spp. The ant nest metabolome is further complicated by competition between defensive bacteria, which produce antibacterials active against even closely related species.


The EVEREST study: an international collaboration.
Rates of initiation of renal replacement therapy (RRT), use of home modalities of treatment and patient outcomes vary considerably between countries. This paper reports the methods and baseline characteristics of countries participating in the EVEREST study (n = 46), a global collaboration examining the association between medical and non-medical factors and RRT incidence, modality mix and survival. Numbers of incident and prevalent patients were collected for current (2003-05) and historic (1983-85, 1988-90, 1993-95 and 1998-2000) periods stratified, where available, by age, gender, treatment modality and cause of end stage renal disease (diabetic versus non-diabetic). General population age and health indicators and national-level macroeconomic data were collected from secondary data sources. National experts provided primary data on renal service funding, resources and organization. The median (inter quartile range) RRT incidence per million of the population (pmp) was 130 pmp (102-167 pmp). The general population life expectancy at 60 was 22.1 years (19.7-23.1 years) and 6.9% had diabetes mellitus (5.4-9.0%). Healthcare spending as a percentage of gross domestic product was 8.1% (5.6-9.3%). Countries averaged nine dialysis facilities pmp (4-12 pmp), with 69.0% (43.9-99.0%) owned by the public or private not-for-profit sector. The number of nephrologists ranged from 0.5 to 48 pmp (median 12 pmp). The heterogeneity of EVEREST countries will enable modelling to examine the independent association between medical and non-medical factors on RRT epidemiology.


NopB, a type III secreted protein of Rhizobium sp. strain NGR234, is associated with pilus-like surface appendages.
Rhizobium sp. strain NGR234 possesses a functional type three secretion system (TTSS), through which a number of proteins, called nodulation outer proteins (Nops), are delivered to the outside of the cell. A major constraint to the identification of Nops is their low abundance in the supernatants of NGR234 strains grown in culture. To overcome this limitation, a more sensitive proteomics-based strategy was developed. Secreted proteins from wild-type NGR234 were separated by two-dimensional gel electrophoresis, and the gel was compared to similar gels containing the proteins from a TTSS mutant (NGROmegarhcN). To identify the proteins, spots unique to the NGR234 gels were analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry and the data were compared to the sequence of the symbiotic plasmid of NGR234. A nonpolar mutant of one of these proteins was generated called NopB. NopB is required for Nop secretion but inhibits the interaction with Pachyrhizus tuberosus and augments nodulation of Tephrosia vogelii. Flavonoids and a functional TTSS are required for the formation of some surface appendages on NGR234. In situ immunogold labeling and isolation of these pili showed that they contain NopB.


A Guide to Quantitative Biomarker Assay Development using Whispering Gallery Mode Biosensors.
Whispering gallery mode (WGM) sensors are a class of powerful analytical techniques defined by the measurement of changes in the local refractive index at or near the sensor surface. When functionalized with target-specific capture agents, analyte binding can be measured with very low limits of detection. There are many geometric manifestations of WGM sensors, with chip-integrated silicon photonic devices first commercialized because of the robust, wafer-scale device fabrication, facile optical interrogation, and amenability to the creation of multiplexed sensor arrays. Using these arrays, a number of biomolecular targets have been detected in both label-free and label-enhanced assay formats. For example, sub-picomolar detection limits for multiple cytokines were achieved using an enzymatically enhanced sandwich immunoassay that showed high analyte specificity suitable for detection in complex, clinical matrices. This protocol describes a generalizable approach for the development of quantitative, multiplexed immunoassays using silicon photonic microrings as an example WGM platform. © 2017 by John Wiley & Sons, Inc.


Presynaptic enhancement of inhibitory synaptic transmission by protein kinases A and C in the rat hippocampus in vitro.
The protein kinase C activator phorbol 12,13-dibutyrate (0.5 microM, PDBu) and the protein kinase A activator forskolin (20 microM) each increased evoked monosynaptic inhibitory postsynaptic current (IPSC) amplitude, without affecting its reversal potential, and increased the frequency of miniature IPSCs (mIPSCs), without affecting their amplitude or kinetics, as assessed with whole-cell recording form CA3 pyramidal cells in hippocampal slice cultures. The effects of forskolin and PDBu on both evoked IPSC amplitude and mIPSC frequency were additive and were antagonized by inhibitors of protein kinases A and C, respectively. The kinase activator-induced increases in mIPSC frequency were quantitatively comparable to the increases in evoked IPSC amplitude. The increases in mIPSC frequency were not attenuated by the voltage-dependent calcium channel blocker Cd2+ (100 microM). We conclude that stimulation of protein kinases A and C potentiates hippocampal inhibitory synaptic transmission through independent presynaptic mechanisms of action. Kinase-induced potentiation of spontaneous release does not require modulation of axon terminal Ca2+ channels. This mechanism may also contribute substantially to the potentiation of evoked release.


Memory traces of long-range coordinated oscillations in the sleeping human brain.
Cognition involves coordinated activity across distributed neuronal networks. Neuronal activity during learning triggers cortical plasticity that allows for reorganization of the neuronal network and integration of new information. Animal studies have shown post-learning reactivation of learning-elicited neuronal network activity during subsequent sleep, supporting consolidation of the reorganization. However, no previous studies, to our knowledge, have demonstrated reactivation of specific learning-elicited long-range functional connectivity during sleep in humans. We here show reactivation of learning-induced long-range synchronization of magnetoencephalography power fluctuations in human sleep. Visuomotor learning elicited a specific profile of long-range cortico-cortical synchronization of slow (0.1 Hz) fluctuations in beta band (12-30 Hz) power. The parieto-occipital part of this synchronization profile reappeared in delta band (1-3.5 Hz) power fluctuations during subsequent sleep, but not during the intervening wakefulness period. Individual differences in the reactivated synchronization predicted postsleep performance improvement. The presleep resting-state synchronization profile was not reactivated during sleep. The findings demonstrate reactivation of long-range coordination of neuronal activity in humans, more specifically of reactivation of coupling of infra-slow fluctuations in oscillatory power. The spatiotemporal profile of delta power fluctuations during sleep may subserve memory consolidation by echoing coordinated activation elicited by prior learning.


Incidence, Risk Factors, and Clinical Characteristics of Peripartum Cardiomyopathy in South Korea.
Peripartum cardiomyopathy (PPCM) is a rare disorder associated with pregnancy that can lead to life-threatening conditions. The incidence and clinical characteristics of this condition remain poorly understood. We aimed to perform the first population-based study of PPCM in South Korea, using the Korea National Health Insurance Claims Database of the Health Insurance Review and Assessment Service. Patients who fulfilled predefined diagnostic criteria for PPCM from January 1, 2010, to December 31, 2012, were identified from International Classification of Diseases, Tenth Revision, Clinical Modification codes. To discriminate PPCM from other causes of heart failure, we excluded subjects who already had heart failure-related International Classification of Diseases, Tenth Revision, Clinical Modification codes at least 1 year before delivery. During the study period, there were 1 404 551 deliveries in South Korea, and we excluded 20 159 patients who already had heart failure. In those, a total of 795 cases were identified as PPCM. Patients with PPCM were older, had a higher prevalence of preeclampsia and gestational diabetes mellitus, and were more likely to be primiparous and have multiple pregnancies. Moreover, cesarean section and pregnancy-related complications and in-hospital death were also more common in patients with PPCM. Intriguingly, a considerable number of heart failure cases (n=64; 8.1% of total PPCM) were noted between 5 and 12 months after delivery. The incidence of PPCM was 1 in 1741 deliveries in South Korea. Patients with PPCM were older, were more associated with primiparity and multiple pregnancy, had more pregnancy-related complications, and revealed higher in-hospital mortality than controls. The number of cases diagnosed as PPCM were decreased over time after delivery; however, a large number of patients were still noted through 12 months after delivery.


A method to assess endocardial regional longitudinal curvature of the left ventricle.
Knowledge of the instantaneous geometry of the left ventricular (LV) chamber is necessary to calculate LV function and wall stresses. We describe a method utilizing myocardial markers that does not rely on any a priori assumptions of global LV geometry. Five dogs underwent placement of 25 endocardial and 3 epicardial miniature LV markers. Six weeks later, the animals were studied during conscious closed-chest conditions. The three-dimensional coordinates of the LV markers were used to compute longitudinal fitted curves for LV walls and septum during steady-state conditions; endocardial radii of curvature (rcurv) were then computed for each region at the midequatorial (rcurv-eq) and apical levels. There was a uniform decrease in rcurv in each LV wall during systole (compared with diastole, P < 0.01); at end systole, rcurv was regionally heterogeneous between opposing walls, e.g., anterior and posterior rcurv-eq values were 17.2 +/- 2.0 and 17.7 +/- 1.8 (SD) cm, respectively (P < 0.05). At end diastole, only septal-lateral rcurv-eq was different (16.9 +/- 2.1 vs. 18.7 +/- 1.3 cm: P < 0.05). Normalization of rcurv (to instantaneous LV volume) removed the systolic-diastolic differences, but a similar pattern of regional heterogeneity persisted. The data presented pertain to the LV endocardial surface, but the method described can be applied to the epicardial surface as well; this new method offers promise in assessing dynamic changes in longitudinal LV endocardial curvature.


Modulation of Lipid Droplet Metabolism-A Potential Target for Therapeutic Intervention in Flaviviridae Infections.
Lipid droplets (LDs) are endoplasmic reticulum (ER)-related dynamic organelles that store and regulate fatty acids and neutral lipids. They play a central role in cellular energy storage, lipid metabolism and cellular homeostasis. It has become evident that viruses have co-evolved in order to exploit host lipid metabolic pathways. This is especially characteristic of the Flaviviridae family, including hepatitis C virus (HCV) and several flaviviruses. Devoid of an appropriate lipid biosynthetic machinery of their own, these single-strand positive-sense RNA viruses can induce dramatic changes in host metabolic pathways to establish a favorable environment for viral multiplication and acquire essential components to facilitate their assembly and traffic. Here we have reviewed the current knowledge on the intracellular life cycle of those from the Flaviviridae family, with particular emphasis on HCV and dengue virus (DENV), and their association with the biosynthesis and metabolism of LDs, with the aim to identify potential antiviral targets for development of novel therapeutic interventions.


Natural history of frontotemporal dementia: comparison with Alzheimer's disease.
Frontotemporal dementia (FTD) is a more common cause of dementia than previously recognised. Few data are available regarding the natural course of FTD in terms of survival, nursing home admission and causes of death. An observational study of all consecutive patients referred to the memory centre of Lille, France, between 1995 and 1999, and examined at least twice in this centre, with a diagnosis of FTD (frontal or behavioural variant) or of Alzheimer's disease (AD) was performed. Kaplan-Meyer analysis allowing for delayed entry was used to compare the survival functions in FTD and AD. 552 patients were included, of whom 49 (8.9%) were lost to follow-up at 3 years. FTD patients were younger (mean age at onset 59 years), had more often a family history of psychiatric disorders (20%), had a longer delay between first symptoms and first visit (5.9 years) and a higher Mini-Mental State Examination (MMSE) score at first visit (24.5) than patients with AD (19.9). The mean annual MMSE score decline was 0.9 point in FTD vs. 2.0 points in AD (p < 0.0004). Fewer patients with FTD than with AD entered an institution (RR: 0.20, 95% CI 0.05-0.81). After adjustment for sex, age at first visit, level of education and MMSE score at first visit, survival rates in FTD and AD did not differ significantly. Patients with FTD often had a sudden death, the cause of which could not be found. The earlier the first visit after onset, the longer the survival rate, whatever the diagnosis (RR: 0.76, 95% CI 0.67-0.86, p < 0.0001 per year of earlier first visit). This large study showed that the mean duration of FTD was 2 years longer than that of AD, but the risk of death after adjustment for age and sex was similar in FTD and in AD. Sudden and unexplained causes of death were frequent and need further study. Early management increases the life span of demented patients.


Tumour-necrosis-factor-mediated cytotoxicity is correlated with phospholipase-A2 activity, but not with arachidonic acid release per se.
L929, a murine fibrosarcoma cell line highly sensitive to the anti-proliferative and cytotoxic action of tumour necrosis factor (TNF), was used as a target cell in our studies. We [Suffys et al. (1987) Biochem. Biophys. Res. Commun. 149, 735-743], as well as others, have previously provided evidence that a phospholipase (PL), most probably a PL-A2-type enzyme, is likely to be involved in TNF-mediated cell killing. We now further document this conclusion and provide suggestive evidence that the enzyme activity specifically involved in TNF cytotoxicity differs from activities associated with the eventual cell death process itself or with non-toxic serum treatment. We also show that the 5,8,11,14-icosatetraenoic acid (arachidonic acid, delta 4 Ach) released by PL, and possibly metabolized, is unlikely to be a key mediator of the TNF-mediated cytotoxicity. These conclusions are based on the following experimental findings. 1. TNF treatment of cells, prelabelled for 24 h with [3H] delta 4Ach or [14C] delta 3Ach (delta 3Ach identical to 5,8,11-icosatrienoic acid) resulted in an early, time-dependent and concentration-dependent release of radioactivity in the supernatant preceding actual cell death. The extent of this response was moderate, albeit reproducible and significant. Analysis of the total lipid fraction from cells plus supernatant revealed that only release of arachidonic acid from phospholipids, but not its metabolization was induced by TNF. However, the release of less unsaturated fatty acids, such as linoleic acid (Lin) or palmitic acid (Pam), was not affected during the first hours after TNF addition. 2. An L929 subclone, selected for resistance to TNF toxicity, was found to be defective in TNF-induced delta 4Ach libration. 3. Interleukin-1 (IL1) was not cytotoxic for L929 and did not induce release of delta 4Ach. 4. Release of delta 4Ach was not restricted to TNF; the addition of serum to the cells also induced release of fatty acids into the medium. In this case, however, there was no specificity, as all fatty acids tested, including Lin and Pam, were released. 5. Inhibition of PL-A2 activity by appropriate drugs markedly diminished TNF-induced delta 4Ach release and resulted also in a strong decrease in TNF-induced cytotoxicity. 6. Other drugs, including serine protease inhibitors, which strongly inhibit TNF-induced cytotoxicity, also decreased the TNF-induced delta 4Ach release, whereas LiCl potentiated both TNF-mediated effects. 7. Protection of cells against TNF toxicity by means of various inhibitors was not counteracted by addition of exogenous fatty acids, including delta 4Ach.(ABSTRACT TRUNCATED AT 400 WORDS)


Exercise performance in children with asthma: is it different from that of healthy controls?
Exercise tolerance and possible limitation in work capacity of asthmatic children is still a matter of debate. The aim of this study was to compare ventilation and gas exchange response to exercise of asthmatic children with that of healthy controls. Exercise performance was evaluated in 80 children with mild-to-moderate asthma, aged 7-15 yrs, and in 80 healthy controls matched for age, height, weight and habitual level of physical activity. The children performed a maximal exercise test on a treadmill, during which oxygen uptake (V'O2), carbon dioxide output (V'CO2) and minute ventilation (V'E) were measured continuously. No premedication was given to the asthmatic children. Forced expiratory volume in one second (FEV1) at rest was 93+/-11% of predicted in asthmatic children and 95+/-9% pred in controls. After the run, the mean fall in FEV1 was 13.9% (range 0-57%) and 1.6% (0-9%), respectively (p<0.001). The two groups achieved similar maximum oxygen uptake (V'O2,max) ((mean+/-SD) 40.3+/-8.4 and 42.6+/-9.6 mL x min(-1) x kg(-1) in asthmatics and controls, respectively; NS) and maximum minute ventilation output (V'E,max) (42.9+/-14.8 and 45.7+/-14.9 L x min(-1) respectively; NS). The kinetics of V'O2, V'CO2 and V'E during the test revealed no differences between the two populations. Moreover, anaerobic threshold and oxygen pulse were the same in the two groups. Asthmatics showed a ventilatory pattern with lower respiratory frequencies and greater tidal volumes during the run. These results suggest that asthmatic children can achieve a level of exercise performance similar to that of healthy children, provided that they have a comparable level of habitual physical activity. The only difference found concerned the ventilatory pattern of the asthmatic children, which was characterized by a reduced respiratory frequency and greater tidal volume at the same minute ventilation. The level of physical conditioning was found to be the main determinant of exercise tolerance for children with controlled asthma.


The effect of brevenal on brevetoxin-induced DNA damage in human lymphocytes.
Brevenal is a nontoxic short-chain trans-syn polyether that competes with brevetoxin (PbTx) for the active site on voltage-sensitive sodium channels. The PbTxs are highly potent polyether toxins produced during blooms of several species of marine dinoflagellates, most notably Karenia brevis. Blooms of K. brevis have been associated with massive fish kills, marine mammal poisoning, and are potentially responsible for adverse human health effects such as respiratory irritation and airway constriction in beach-goers. Additionally, the consumption of shellfish contaminated with PbTxs results in neurotoxic shellfish poisoning (NSP). The purpose of the present study was to determine whether PbTx could induce DNA damage in a human cell type, the lymphocyte, and if so, whether the damage could be antagonized or ameliorated by brevenal, a brevetoxin antagonist. The DNA damage may occur through both endogenous and exogenous physiological and pathophysiological processes. Unrepaired or erroneously repaired DNA damage may result in gene mutation, chromosome aberration, and modulation of gene regulation, which have been associated with immunotoxicity and carcinogenesis. A single-cell gel electrophoresis assay, or comet assay, was used to determine and compare DNA damage following various treatments. The data were expressed as tail moments, which is the percentage of DNA in the tail multiplied by the length between the center of the head and center of the tail (in arbitrary units). The negative control tail moment was 29.2 (SE=+/-0.9), whereas the positive control (hydrogen peroxide) was 72.1 (1.5) and solvent (ethanol) was 24.2 (2.1). The PbTx-2 (from Sigma, St. Louis, MO, USA), 10(-8) M was 41.3 (3.6), PbTx-9 (Sigma), 10(-8) M was 57.0 (5.3), PbTx-2 (from University of North Carolina at Wilmington, UNCW), 10(-8) M was 49.4 (9.9), and PbTx-3 (UNCW), 10(-8) M was 64.0 (6.4). 1.0 microg/ml brevenal applied 1 h before the PbTxs protected the lymphocytes from DNA damage; PbTx-2 (Sigma), 31.3 (2.1); PbTx-9 (Sigma), 35.5 (2.9); PbTx-2 (UNCW), 33.9 (1.4); PbTx-3 (UNCW), 34.9 (1.25). The tail moment for 1.0 mug/ml brevenal alone was 30.8 (2.6). The results indicate that extensive genotoxic damage is induced by PbTx-2 and 9 (Sigma), and PbTx-2 and 3 (UNCW) in normal human lymphocytes, which is fully antagonized by brevenal. This suggests that the immune systems of individuals exposed to PbTx during harmful algal bloom (HAB) events may be at risk.


Factors influencing clinical decisions to initiate thyroxine therapy for patients with mildly increased serum thyrotropin (5.1-10.0 mIU/L).
To determine factors that influence clinicians' decisions to treat subclinical hypothyroidism. We studied patients living within 120 miles of our institution in Rochester, Minn, whose serum thyrotropin value was between 5.1 and 10.0 mIU/L from January 1, 1995, through December 31,1996. Exclusion criteria were history of thyroid disease or prior abnormal thyrotropin level and fewer than 5 follow-up visits or fewer than 2 follow-up thyrotropin measurements. Of the 2655 patients in this cohort, only 55% (1466) had measurement of thyroid microsomal antibodies (TMAb). From this group, records of 539 patients were selected randomly according to antibody status (238 with positive, 199 with negative, and 102 with no TMAb measurement). After exclusion of patients receiving thyroxine replacement and other medications affecting thyroid function, patients with postpartum thyroiditis, and patients in the intensive care unit, data from 450 patients were analyzed. Thyroxine therapy was prescribed for 39% of the patients with thyrotropin levels between 5.1 and 10.0 mIU/L. In multivariate analysis, TMAb status (P < .001), index thyrotropin (P < .002), and free thyroxine (FT4) (P = .03) were the strongest predictors for the decision to treat. Fifty-nine percent of the patients with positive TMAb were treated compared with 24% of those with negative TMAb and 30% of those who did not have a TMAb test. The mean thyrotropin level in the treated group was 7.1 mIU/L compared with 63 mIU/L in the untreated group. The mean FT4 level was 1.08 ng/dL in the treated group who had this measured compared with 1.16 ng/dL in the untreated group. Treated patients were younger than untreated patients (56.5 years vs 60.6 years), and the proportion of female patients was higher in the treated group compared with the untreated group (75.8% vs 65.2%); however, neither of these differences was significant after correction for the multiple comparisons. In multivariate analysis neither age nor sex was a significant predictor of treatment, but in the subgroup comparison, patients aged 31 to 50 years were twice as likely to be treated compared with those aged 61 to 80 years. Among patients with serum thyrotropin levels of 5.1 to 10.0 mIU/L, those with positive TMAb values, those with higher thyrotropin values, and those with lower FT4 values were more likely to receive thyroxine therapy. Younger patients were more likely to be treated than older patients.


Rational design, synthesis, and biological evaluation of Pan-Raf inhibitors to overcome resistance.
Selective BRafV600E inhibitors with DFG-in conformation have been proven effective against a subset of melanoma. However, representative inhibitor vemurafenib rapidly acquires resistance in the BRafWT cells through a CRaf or BRafWT dependent manner. Simultaneous targeting of all subtypes of Raf proteins offers the prospect of enhanced efficacy as well as reduced potential for acquired resistance. Herein, we describe the design and characterization of a series of compounds I-01-I-22, based on a pyrimidine scaffold with DFG-out conformation as Pan-Raf inhibitors. Among them, I-15 binds to all Raf protomers with IC50 values of 12.6 nM (BRafV600E), 30.1 nM (ARaf), 19.7 nM (BRafWT) and 17.5 nM (CRaf) and demonstrates cellular activity against BRafWT phenotypic melanoma and BRafV600E phenotypic colorectal cancer cells. The western blot results for the P-Erk inhibition in human melanoma SK-Mel-2 cell line showed that I-15 inhibited the proliferation of the SK-Mel-2 cell line at concentrations as low as 400 nM, without paradoxical activation of Erk as vemurafenib, which supported that I-15 may become a good candidate compound to overcome the resistance of melanoma induced by vemurafenib. I-15 also has a favorable pharmacokinetic profile in rats. Rational design, synthesis, SAR, lead selection and evaluation of the key compounds studied are described.


In situ butanol recovery from Clostridium acetobutylicum fermentations by expanded bed adsorption.
Although butanol is a promising biofuel, its fermentative production suffers from inhibition caused by end product toxicity. The in situ removal of butanol from cultures via expanded bed adsorption offers an effective strategy for mitigating the effects of product toxicity while eliminating the need to clarify cultures via microfiltration. The hydrophobic polymer resin Dowex Optipore L-493 was found to be both an effective butanol adsorbent and suitable for use in expanded bed adsorption. Recirculation rates through the adsorption column were strongly correlated with and ultimately controlled rates of butanol uptake from the media which, reaching as high as 41.1 g/L h, easily exceed those of its production in a typical fermentation. Vacuum application with vapor collection was found to be an effective means of adsorbent regeneration, with an average of 81% butanol recovery possible, with butanol concentrations in the cold trap reaching as high as 85.8 g/L. Integration of expanded bed adsorption with a fed-batch Clostridium acetobutylicum ATCC 824 fermentation and its continuous operation for 38.5 h enabled the net production (i.e., in solution and adsorbed) of butanol and total solvent products at up to 27.2 and 40.7 g/L of culture, respectively, representing 2.2- and 2.3-fold improvements over conventional batch culture. While adsorbent biofouling was found to be minimal, further investigation of biofouling in longer-term studies will provide useful and further insight regarding the robustness of the process strategy.


Family caregiver burden: results of a longitudinal study of breast cancer patients and their principal caregivers.
The vital role played by family caregivers in supporting dying cancer patients is well recognized, but the burden and economic impact on caregivers is poorly understood. We prospectively examined the psychosocial, occupational and economic impact of caring for a person with a terminal illness. We studied 89 caregivers of women with advanced breast cancer receiving care at either the Ottawa or Hamilton regional cancer centres in Ontario. Patients were followed until their death or study completion at 3 years. Patients identified a principal caregiver to participate in the study. The Karnofsky Performance Status (KPS) index, the Medical Outcomes Study 36-item Short Form (SF-36), the Hospital Anxiety and Depression Scale, the Zarit Burden Inventory, FAMCARE and the Medical Outcomes Study Social Support Survey were administered during follow-up. Economic data were collected by means of a questionnaire administered by an interviewer. Assessments were conducted every 3 months during the palliative period (KPS score > 50) and every 2 weeks during the terminal period (KPS score < or = 50). Over half of the caregivers were male (55%) and the patient's spouse or partner (52%), with a mean age of 53 years. At the start of the palliative period, the caregivers' mean physical functioning score was better than the patients' (51.3 v. 35.1, 95% confidence interval [CI] 13.3-20.0); there were similar mean mental functioning scores (46.6 and 47.1 respectively); similar proportions were depressed (11% and 12%); and significantly more caregivers than patients were anxious (35% v. 19%, p = 0.009). More caregivers were depressed (30% v. 9%, p = 0.02) and had a higher level of perceived burden (26.2 v. 19.4, p = 0.02) at the start of the terminal period than at the start of the palliative period. Burden was the most important predictor of both anxiety and depression. Of employed caregivers, 69% reported some form of adverse impact on work. In the terminal period 77% reported missing work because of caregiving responsibilities. Prescription drugs were the most important component of financial burden. Caregivers' depression and perceived burden increase as patients' functional status declines. Strategies are needed to help reduce the psychosocial, occupational and economic burden associated with caregiving.


Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer.
Gap junctions are intercellular channels composed of connexins, which mediate the direct passage of small molecules between neighbouring cells. They are involved in regulation of cell cycle, cell signalling, and differentiation, and probably invasion and metastasis. The role of connexins in the metastatic process is controversial, because some studies indicate that connexin expression is inversely correlated with metastatic capacity. In contrast, others demonstrate that connexins may be involved in metastasis. In addition, connexin status in breast cancer metastasis has not been widely studied. We evaluated by immunohistochemistry the expression of connexin 26 (Cx26) and connexin 43 (Cx43) in primary breast tumours (PTs) and matched paired metastases to lymph nodes (MLNs). In PTs, we observed predominantly cytoplasmic localisation of evaluated connexins, indicating alterations in connexin expression in breast cancer cells. We demonstrated that expression of Cx26 and Cx43 was increased in MLNs compared with PTs (p<0.00001 and p<0.001, for CX26 and Cx43, respectively). In addition, Cx26 and Cx43 negative PTs developed Cx26 and Cx43 positive MLNs. Furthermore, besides increased cytoplasmic staining, enhanced membranous localisation of Cx43, typical of normal cells, was found in MLNs. Additionally, membranous Cx26 expression appeared only in metastatic breast cancer cells. These findings suggest that connexins may contribute to the efficient metastasising of breast cancer to the lymph nodes.


Intraarticular osteoid osteoma: clinical features, imaging results, and comparison with extraarticular localization.
Intraarticular osteoid osteoma is uncommon and presents diagnostic difficulties, which are important for both rheumatologists and orthopedic surgeons. Clinical symptoms, imaging procedures, differential diagnostic problems, and treatment results of intraarticular as compared with extraarticular osteoma are analyzed in this retrospective study. Nineteen patients with intraarticular osteoid osteomas (Group A), with a mean followup period of 34 months, are compared with 15 others with extraarticular lesions (Group B). Nine intraarticular tumors were located in the hip, 3 in the elbow, 6 in the ankle, and one in the first metatarsal head. The nonspecific symptoms in Group A, such as chronic synovitis, decreased range of motion, joint effusion, contractures, and lack of the intense perifocal sclerotic margin on radiographs, led to significant delay in diagnosis (on average 26.6 mo in Group A, 8.5 mo in Group B). The extreme variety of previous diagnoses at referral reflect the problems of differential diagnosis. A detectable nidus is often absent on conventional radiograph. Bone scintigraphy is unspecific and often fails to visualize the nidus. Computed tomography scans were accurate in two-thirds of the intraarticular and in 90% of extraarticular cases. Magnetic resonance image findings, although sometimes controversial, provided essential additional information for the correct diagnosis and therapy. Clinical symptoms and imaging signs of intraarticular osteoid osteomas were significantly different from the classical hallmarks of extraarticular lesions. The 10% intraarticular occurrence of osteoid osteomas in this series is not as rare as some investigators suggest. The radiological and clinical findings are uncharacteristic and misleading, and the lesions are difficult to identify. Careful search for history data, such as nocturnal pain and positive salicylate test, in addition to extensive imaging procedures, led to the correct diagnosis prior to surgery in two-thirds of our patients with intraarticular osteoid osteomas.


An Update on the Outcomes of Patients Treated with Urolift for Benign Prostatic Hyperplasia.
Benign prostatic hyperplasia (BPH) is a ubiquitous urologic disease affecting aging men. Patients often experience bothersome lower urinary tract symptoms (LUTS) that warrant urologic evaluation and management. Routinely, patients are initially treated with medical therapies with the goal of both relaxing the bladder neck and shrinking the prostate in order to relieve obstruction secondary to prostatic enlargement. Transurethral resection of the prostate (TURP) serves as a first-line surgical intervention in those who fail medical therapy. Recently, novel minimally invasive surgical techniques for BPH management have emerged. Of these, prostatic urethral lift (PUL or Urolift) has gained attention given its presumed effectiveness and minimal risk of sexual side effects when compared to the standard TURP. The purpose of this review is to describe past and current trends in the implementation of PUL for BPH and to highlight important outcomes.


Beyond inhibitory control training: Inactions and actions influence smartphone app use through changes in explicit liking.
Human behavior can be classified into 2 basic categories: execution of responses and withholding responses. This classification is used in go/no-go training, where people respond to some objects and withhold their responses to other objects. Despite its simplicity, there is now substantial evidence that such training is powerful in changing human behavior toward such objects. However, it is poorly understood how simple responses can influence behavior. Contrary to the remarkably tenacious idea that go/no-go training changes behavior by strengthening inhibitory control, we propose that the training changes behavior via changes in explicit liking of objects. In two preregistered experiments, we show that go/no-go training influences explicit liking for smartphone apps (Experiments 1 and 2) and that this liking partially mediates the effect of the training on consequential choices for using these apps 1 day later (Experiment 2). The results highlight the role of evaluations when examining how motor response training influences behavior. This knowledge can inform development of more effective applied motor response training procedures and raises new theoretical questions on the relation between motor responses and affect. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


A description of interventions promoting healthier ready-to-eat meals (to eat in, to take away, or to be delivered) sold by specific food outlets in England: a systematic mapping and evidence synthesis.
Ready-to-eat meals (to eat in, to take away or to be delivered) sold by food outlets are often more energy dense and nutrient poor compared with meals prepared at home, making them a reasonable target for public health intervention. The aim of the research presented in this paper was to systematically identify and describe interventions to promote healthier ready-to-eat meals (to eat in, to take away, or to be delivered) sold by specific food outlets in England. A systematic search and sift of the literature, followed by evidence mapping of relevant interventions, was conducted. Food outlets were included if they were located in England, were openly accessible to the public and, as their main business, sold ready-to-eat meals. Academic databases and grey literature were searched. Also, local authorities in England, topic experts, and key health professionals and workers were contacted. Two tiers of evidence synthesis took place: type, content and delivery of each intervention were summarised (Tier 1) and for those interventions that had been evaluated, a narrative synthesis was conducted (Tier 2). A total of 75 interventions were identified, the most popular being awards. Businesses were more likely to engage with cost neutral interventions which offered imperceptible changes to price, palatability and portion size. Few interventions involved working upstream with suppliers of food, the generation of customer demand, the exploration of competition effects, and/or reducing portion sizes. Evaluations of interventions were generally limited in scope and of low methodological quality, and many were simple assessments of acceptability. Many interventions promoting healthier ready-to-eat meals (to eat in, to take away, or to be delivered) sold by specific food outlets in England are taking place; award-type interventions are the most common. Proprietors of food outlets in England that, as their main business, sell ready-to-eat meals, can be engaged in implementing interventions to promote healthier ready-to-eat-food. These proprietors are generally positive about such interventions, particularly when they are cost neutral and use a health by stealth approach.


Implication of common and disease specific variants in CLU, CR1, and PICALM.
Two recent genome-wide association studies (GWAS) for late onset Alzheimer's disease (LOAD) revealed 3 new genes: clusterin (CLU), phosphatidylinositol binding clathrin assembly protein (PICALM), and complement receptor 1 (CR1). In order to evaluate association with these genome-wide association study-identified genes and to isolate the variants contributing to the pathogenesis of LOAD, we genotyped the top single nucleotide polymorphisms (SNPs), rs11136000 (CLU), rs3818361 (CR1), and rs3851179 (PICALM), and sequenced the entire coding regions of these genes in our cohort of 342 LOAD patients and 277 control subjects. We confirmed the association of rs3851179 (PICALM) (p = 7.4 × 10(-3)) with the disease status. Through sequencing we identified 18 variants in CLU, 3 of which were found exclusively in patients; 8 variants (out of 65) in CR1 gene were only found in patients and the 16 variants identified in PICALM gene were present in both patients and controls. In silico analysis of the variants in PICALM did not predict any damaging effect on the protein. The haplotype analysis of the variants in each gene predicted a common haplotype when the 3 single nucleotide polymorphisms rs11136000 (CLU), rs3818361 (CR1), and rs3851179 (PICALM), respectively, were included. For each gene the haplotype structure and size differed between patients and controls. In conclusion, we confirmed association of CLU, CR1, and PICALM genes with the disease status in our cohort through identification of a number of disease-specific variants among patients through the sequencing of the coding region of these genes.


Psychometric properties of the Korean version of the Health Literacy on Social Determinants of Health Questionnaire (K-HL-SDHQ).
The association between the social determinant of health (SDH) and sustainable development goals, has directed attention toward the influence of SDH. However, there is a lack of evidence regarding the instruments used to assess SDH. Thus, this study was conducted to assess the validity and reliability of the Korean Version of the Health Literacy on Social Determinants of Health Questionnaire (K-HL-SDHQ). A total of 660 workers in Korea participated in an online survey. The K-HL-SDHQ measures four dimensions (Access, Understand, Appraise, and Apply) with 33 items. The HL-SDHQ was translated into Korean using the forward-back translation method. To test the validity and reliability of the Korean translated HL-SDHQ, item analysis for the 33 items was conducted. Internal consistency was examined using Cronbach's α, an exploratory factor analysis, and a confirmatory factor analysis. The scale-level content validity index (S-CVI)/universal agreement of this study was .12 and S-CVI/average was .83 (item-CVI range = .50-1.00). The goodness of fit determined through a confirmatory factor analysis of the four dimensions was acceptable (χ2 (489) = 1475.054, p < .001, RMSEA = .06, CFI = .87, TLI = .85). The K-HL-SDHQ also demonstrated satisfactory internal consistency reliability (Cronbach's α = .92). The findings indicate that the K-HL-SDHQ is a valid and reliable tool that can be used to assess the SDH of workers in Korea. It is suggested that this tool can be applied through repeated research with workers and non-workers for health promotion, and to enhance researchers' understanding of the different levels of the HL-SDHQ.


An investigation of scavenger receptor A mediated leukocyte binding to polyanionic and uncharged polymer hydrogels.
Cell adhesion to biomaterials can be mediated in part by mechanisms aside from the traditionally recognized opsinization and integrin binding mechanisms. In this study, we investigated the role of scavenger receptor A (SR-A) in leukocyte binding to a series of well-controlled polyanionic and uncharged hydrogels based on a poly(N-isopropylacrylamide) backbone. The hydrogels were injected in the peritoneal cavity of SR-A knockout (KO) and wild-type mice using a minimally invasive procedure and allowed to set in situ. After 24 h, the hydrogels were recovered and analyzed, the peritoneal cavity was lavaged, and cytokine concentrations were assessed by ELISA. The polyanionic hydrogels retrieved from the KO animals were found to be completely devoid of adherent leukocytes, which were present in other materials regardless of the mouse strain in which they were injected. Results from a subsequent in vitro cellular adhesion study with a RAW264.7 cell line failed to yield a similarly definitive role for SR-A in the cellular binding of a polyanionic hydrogel. Taken together, the results of this study show that SR-A mediates leukocyte adhesion to a polyanionic hydrogel in the peritoneal cavity, but other adhesion mechanisms contribute to cellular binding in vitro.


Cutaneous tuberculosis of the penis and sexual transmission of tuberculosis confirmed by molecular typing.
A case of culture-positive primary cutaneous Mycobacterium tuberculosis infection of the penis was diagnosed in a male patient; 1 year later, endometrial tuberculosis was diagnosed in the patient's wife. These organisms were confirmed to be indistinguishable by use of molecular techniques.


Identification of miRNAs and their target genes in developing maize ears by combined small RNA and degradome sequencing.
In plants, microRNAs (miRNAs) are endogenous ~22 nt RNAs that play important regulatory roles in many aspects of plant biology, including metabolism, hormone response, epigenetic control of transposable elements, and stress response. Extensive studies of miRNAs have been performed in model plants such as rice and Arabidopsis thaliana. In maize, most miRNAs and their target genes were analyzed and identified by clearly different treatments, such as response to low nitrate, salt and drought stress. However, little is known about miRNAs involved in maize ear development. The objective of this study is to identify conserved and novel miRNAs and their target genes by combined small RNA and degradome sequencing at four inflorescence developmental stages. We used deep-sequencing, miRNA microarray assays and computational methods to identify, profile, and describe conserved and non-conserved miRNAs at four ear developmental stages, which resulted in identification of 22 conserved and 21-maize-specific miRNA families together with their corresponding miRNA*. Comparison of miRNA expression in these developmental stages revealed 18 differentially expressed miRNA families. Finally, a total of 141 genes (251 transcripts) targeted by 102 small RNAs including 98 miRNAs and 4 ta-siRNAs were identified by genomic-scale high-throughput sequencing of miRNA cleaved mRNAs. Moreover, the differentially expressed miRNAs-mediated pathways that regulate the development of ears were discussed. This study confirmed 22 conserved miRNA families and discovered 26 novel miRNAs in maize. Moreover, we identified 141 target genes of known and new miRNAs and ta-siRNAs. Of these, 72 genes (117 transcripts) targeted by 62 differentially expressed miRNAs may attribute to the development of maize ears. Identification and characterization of these important classes of regulatory genes in maize may improve our understanding of molecular mechanisms controlling ear development.


Natural history and classification of occlusive thromboaortopathy (Takayasu's disease).
Fifty-four Japanese patients with occlusive thromboaortopathy (OTAP), including four males, were classified according to evidence of complications attributed to OTAP at the time when the diagnosis was established: Group I, uncomplicated OTAP with or without the involvement of the pulmonary artery. Group II, mono-complicated OTAP: presence of a single complication together with uncomplicated OTAP. Group II was subdivided according to severity into group IIa - mild or moderate form, and group IIb - severe form. Group III, multi-complicated OTAP with two or more complications as well as uncomplicated OTAP. The five-year survival rate after established diagnosis was 83.1%. Seven patients died of OTAP within five years, but all had belonged to group IIb or III.