Effects of Intermittent Pneumatic Compression on Leg Vascular Function in People with Spinal Cord Injury: A Pilot Study.
Objective: The purpose of this pilot study was to determine whether 60 mins of intermittent pneumatic compression therapy (IPC) could acutely increase leg blood flow-induced shear stress and enhance vascular endothelial function in persons with spinal cord injury (SCI). Design: Pretest with multiple posttests, within subject randomized control design. Setting: University of Southern Mississippi, Spinal Cord Injury Research Program within the School of Kinesiology, recruiting from the local community in Hattiesburg, Jackson, and Gulfport, MS. Participants: Eight adults with SCI (injury level: T3 and below; ASIA class A-C; age: 41±17 yrs). Interventions: A 60-min IPC session was performed in one leg (experimental leg; EXP), with the other leg serving as a control (CON). Outcomes Measures: Posterior-tibial artery shear rate (Doppler-ultrasound) was examined at rest, and at 15 and 45 mins during IPC. Endothelial function was assessed using the flow-mediated dilation (FMD) technique, before and after IPC. Results: Resting FMD (mm) was similar between legs at rest. A two-way repeated measures ANOVA (leg x time) revealed that during IPC, peak shear rate increased in the EXP leg (215±137 to 285±164 s-1 at 15 mins; +39±29%, P = 0.03), with no change occurring in the CON. In addition, FMD significantly increased in the EXP leg (Pre IPC: 0.36±0.14 vs. Post IPC: 0.47±0.17 mm; P = 0.011, d = 0.66), with no change occurring in the CON leg. Conclusion: These preliminary findings suggests that IPC therapy may acutely increase leg shear stress within 15 mins, with a resultant moderate-large improvement in vascular endothelial function after 60 mins in people with SCI.


Comparison of treatment of patients with congestive heart failure by cardiologists versus noncardiologists.
The frequency of treatment with Heart Failure Society of America (HFSA)-recommended medications for the management of congestive heart failure (CHF) by cardiologists versus noncardiologists was studied. The medical records of 227 patients admitted to our institution between January and June 2000 with a discharge diagnosis of CHF were retrospectively reviewed. Data collected included demographic information and medical history, severity of illness, prescribed level of hospital care at the time of admission, diagnostic evaluations conducted during admission, medications prescribed at admission and discharge, and any noted contraindications to these medications. Data for patients treated by cardiologists versus noncardiologists were compared. Patients treated by cardiologists were significantly more likely to be admitted to an intensive care unit; receive chest x-rays, electrocardiograms, nuclear medicine tests, cardiac catheterizations, and stress tests; and have their weight monitored daily than were patients treated by noncardiologists. The majority of patients with CHF who were eligible for an angiotensin-converting-enzyme (ACE) inhibitor, a beta-blocker, or an aldosterone antagonist did not receive these medications, regardless of the treating physician. Cardiologists prescribed significantly more beta-blockers and aldosterone antagonists for eligible patients at hospital admission. Greater differences were seen in discharge medications, as cardiologists were significantly more likely to prescribe ACE inhibitors, digoxin, beta-blockers, and aldosterone antagonists. Hospitalized patients with CHF were more likely to receive HFSA-recommended medications on admission and discharge when treated by cardiologists versus noncardiologists. Neither cardiologists nor noncardiologists prescribed ACE inhibitors to all eligible patients as frequently as recommended by HFSA guidelines.


Mortality and Vascular Events Among Elderly Patients With Chronic Myeloid Leukemia: A Retrospective Analysis of Linked SEER-Medicare Data.
Tyrosine-kinase inhibitors (TKIs) can be associated with vascular events (VEs). The expected VE rates in patients with chronic myeloid leukemia (CML) are unknown. The present study examined the event rates and mortality among elderly patients with and without CML. Linked Surveillance, Epidemiology, and End Results cancer registry and Medicare claims data were used to identify patients aged ≥ 66 years with an incident (index) diagnosis of CML from 2004 to 2009. A comparison cohort of patients without cancer was matched 1:1 to the CML cohort. All patients were followed up from 12 months before the index diagnosis through death or December 31, 2010. The overall survival and rates of myocardial infarction (MI), stroke, pulmonary embolism (PE), and peripheral arterial disease (PAD) were analyzed. A total of 1466 patients with CML (mean age, 78 years; average follow-up period, 25 months) were identified and matched 1:1 to a noncancer cohort (mean age, 78 years; follow-up period, 42 months). Compared with the noncancer patients, those with CML had greater mortality (63% vs. 23% died during the follow-up period; median survival, 23 vs. > 84 months) and greater rates of MI (33.0 vs. 11.9 per 1000 person-years), stroke (83.2 vs. 43.0), PE (6.6 vs. 2.6), and PAD (92.1 vs. 59.3; P < .01 for all). Of the 15% of CML patients with TKI claims, 97% had received imatinib. The event rates were not elevated for TKI-treated patients compared with the overall group of patients with CML. Elderly patients with CML had greater mortality and greater rates of MI, stroke, PE, and PAD than did noncancer patients. The event rates were not elevated among the TKI-treated (primary imatinib) patients, suggesting that the VE risk in these patients with CML was driven primarily by the underlying factors associated with CML.


Rare BANF1 Alleles and Relatively Frequent EMD Alleles Including 'Healthy Lipid' Emerin p.D149H in the ExAC Cohort.
Emerin (EMD) and barrier to autointegration factor 1 (BANF1) each bind A-type lamins (LMNA) as fundamental components of nuclear lamina structure. Mutations in LMNA, EMD and BANF1 are genetically linked to many tissue-specific disorders including Emery-Dreifuss muscular dystrophy and cardiomyopathy (LMNA, EMD), lipodystrophy, insulin resistance and type 2 diabetes (LMNA) and progeria (LMNA, BANF1). To explore human genetic variation in these genes, we analyzed EMD and BANF1 alleles in the Exome Aggregation Consortium (ExAC) cohort of 60,706 unrelated individuals. We identified 13 rare heterozygous BANF1 missense variants (p.T2S, p.H7Y, p.D9N, p.S22R, p.G25E, p.D55N, p.D57Y, p.L63P, p.N70T, p.K72R, p.R75W, p.R75Q, p.G79R), and one homozygous variant (p.D9H). Several variants are known (p.G25E) or predicted (e.g., p.D9H, p.D9N, p.L63P) to perturb BANF1 and warrant further study. Analysis of EMD revealed two previously identified variants associated with adult-onset cardiomyopathy (p.K37del, p.E35K) and one deemed 'benign' in an Emery-Dreifuss patient (p.D149H). Interestingly p.D149H was the most frequent emerin variant in ExAC, identified in 58 individuals (overall allele frequency 0.06645%), of whom 55 were East Asian (allele frequency 0.8297%). Furthermore, p.D149H associated with four 'healthy' traits: reduced triglycerides (-0.336; p = 0.0368), reduced waist circumference (-0.321; p = 0.0486), reduced cholesterol (-0.572; p = 0.000346) and reduced LDL cholesterol (-0.599; p = 0.000272). These traits are distinct from LMNA-associated metabolic disorders and provide the first insight that emerin influences metabolism. We also identified one novel in-frame deletion (p.F39del) and 62 novel emerin missense variants, many of which were relatively frequent and potentially disruptive including p.N91S and p.S143F (∼0.041% and ∼0.034% of non-Finnish Europeans, respectively), p.G156S (∼0.39% of Africans), p.R204G (∼0.18% of Latinx), p.R207P (∼0.08% of South Asians) and p.R221L (∼0.15% of Latinx). Many novel BANF1 variants are predicted to disrupt dimerization or binding to DNA, histones, emerin or A-type lamins. Many novel emerin variants are predicted to disrupt emerin filament dynamics or binding to BANF1, HDAC3, A-type lamins or other partners. These new human variants provide a foundational resource for future studies to test the molecular mechanisms of BANF1 and emerin function, and to understand the link between emerin variant p.D149H and a 'healthy' lipid profile.


The accidental release of exotic species from breeding colonies and zoological collections.
Exotic species have often been introduced into a new country in zoological or botanical gardens or on game and fur farms. When accidentally or deliberately released, these alien species can become invasive and have negative impacts on native plant and animal communities and human activities. This article focuses on a selection of such invasive species: principally the American mink (Neovison vison), but also the coypu (Myocastor coypus), muskrat (Ondatra zibethicus), raccoon dog (Nyctereutes procyonoides), raccoon (Procyon lotor) and African sacred ibis (Threskiornis aethiopicus). In each of these cases, the authors describe the biological characteristics and life history of the species, in relation to its invasive capacity, the origins and establishment of non-native populations, the environmental consequences and possible control measures. The main negative impacts observed are the destruction of habitat, the introduction and/or spread of pathogens and changes in the composition of native communities with consequent effects on biodiversity.


Characterization of Alternaria and Penicillium species from similar substrata based on growth at different temperature, pH and water activity.
Fifty-eight Alternaria isolates representing 10 species or species-groups and 66 Penicillium isolates representing 18 species were examined for their growth response to the combined effects of water activity, temperature and pH in an extended Central Composite Design. Growth responses were recorded as colony diameter after one and two weeks of growth and analysed using different multivariate statistical analyses. The isolates, when analysed by Principal Component Analysis, clustered according to their genus and to some degree to species or species groups and not according to substratum as excepted. Soft Independent Modelling of Class Analogy and Response Surface Analysis showed that growth responses or growth profiles may be used as classification tool. Partial Least Squares Regression showed that a combination of two different media based on Dichloran Rose bengal Yeast Extract Sucrose agar incubated at two different temperatures were enough to get genus segregation and to some extent species segregation. The results also showed that water activity, temperature and pH interact strongly in their effect on growth rates and that the squared products (optima) of water activity, temperature and pH for each isolate were important for modelling the data sufficiently.


MOCCA: a flexible suite for modelling DNA sequence motif occurrence combinatorics.
Cis-regulatory elements (CREs) are DNA sequence segments that regulate gene expression. Among CREs are promoters, enhancers, Boundary Elements (BEs) and Polycomb Response Elements (PREs), all of which are enriched in specific sequence motifs that form particular occurrence landscapes. We have recently introduced a hierarchical machine learning approach (SVM-MOCCA) in which Support Vector Machines (SVMs) are applied on the level of individual motif occurrences, modelling local sequence composition, and then combined for the prediction of whole regulatory elements. We used SVM-MOCCA to predict PREs in Drosophila and found that it was superior to other methods. However, we did not publish a polished implementation of SVM-MOCCA, which can be useful for other researchers, and we only tested SVM-MOCCA with IUPAC motifs and PREs. We here present an expanded suite for modelling CRE sequences in terms of motif occurrence combinatorics-Motif Occurrence Combinatorics Classification Algorithms (MOCCA). MOCCA contains efficient implementations of several modelling methods, including SVM-MOCCA, and a new method, RF-MOCCA, a Random Forest-derivative of SVM-MOCCA. We used SVM-MOCCA and RF-MOCCA to model Drosophila PREs and BEs in cross-validation experiments, making this the first study to model PREs with Random Forests and the first study that applies the hierarchical MOCCA approach to the prediction of BEs. Both models significantly improve generalization to PREs and boundary elements beyond that of previous methods-including 4-spectrum and motif occurrence frequency Support Vector Machines and Random Forests-, with RF-MOCCA yielding the best results. MOCCA is a flexible and powerful suite of tools for the motif-based modelling of CRE sequences in terms of motif composition. MOCCA can be applied to any new CRE modelling problems where motifs have been identified. MOCCA supports IUPAC and Position Weight Matrix (PWM) motifs. For ease of use, MOCCA implements generation of negative training data, and additionally a mode that requires only that the user specifies positives, motifs and a genome. MOCCA is licensed under the MIT license and is available on Github at https://github.com/bjornbredesen/MOCCA .


Full-mode characterization of correlated photon pairs generated in spontaneous downconversion.
Spontaneous parametric downconversion is the primary source to generate entangled photon pairs in quantum photonics laboratories. Depending on the experimental design, the generated photon pairs can be correlated in the frequency spectrum, polarization, position-momentum, and spatial modes. Exploring the spatial modes' correlation has hitherto been limited to the polar coordinates' azimuthal angle, and a few attempts to study Walsh mode's radial states. Here, we study the full-mode correlation, on a Laguerre-Gauss basis, between photon pairs generated in a type-I crystal. Furthermore, we explore the effect of a structured pump beam possessing different spatial modes onto bi-photon spatial correlation. Finally, we use the capability to project over arbitrary spatial mode superpositions to perform the bi-photon state's full quantum tomography in a 16-dimensional subspace.


Prophylactic laparoscopic-assisted total gastrectomy for hereditary diffuse gastric cancer.
Ten percent of gastric cancer (GC) cases are familial, with one third resulting from a mutation in the tumor suppressor gene CDH1. Loss of this important structure can result in hereditary diffuse gastric cancer (HDGC), which carries a high mortality if early diagnosis is not made. Despite its clear genetic origin, optimal management of HDGC family members is controversial, as the utility and efficacy of current cancer screening programs for mutation carriers are unproven. A 53-year-old Caucasian woman was initially seen for genetic screening because multiple family members had mutations of the CDH1 gene. Her pedigree analysis demonstrated 4 generations of gastric cancer, and 2 of the generations carried the CDH1 germline mutation, consistent with HDGC. At endoscopy, the patient's gastric mucosa was normal and random biopsies were also normal. The patient underwent a laparoscopic total gastrectomy. The gross examination of her stomach appeared normal. On histologic examination, however, the stomach was found to have diffuse (signet ring cell) adenocarcinoma in-situ with 11 microscopic foci of invasive adenocarcinoma limited to the lamina propria. Our case is the first reported prophylactic total gastrectomy utilizing a laparoscopic approach, and it highlights the importance of taking a thorough family history and obtaining a pedigree analysis. Endoscopic screening in HDGC cannot rule out diffuse GC, because the stomach and biopsies can be normal despite the presence of adenocarcinoma. Therefore, our case supports the recommendation for prophylactic gastrectomy in HDGC.


Microdialysis of blood from the cardiac venous outflow: a technique for monitoring myocardial ischemia.
To evaluate the possibility of using microdialysis of blood from the great cardiac vein for detecting myocardial ischemia. Microdialysis catheters were placed in the great cardiac vein and the left atrium of pigs for analysis of lactate, glycerol, pyruvate and glucose. Blood samples were drawn for measurement of aspartate aminotransferase, alanine aminotransferase, lactic dehydrogenase and myoglobin with the objective of verifying myocardial damage. Ischemia was induced for 3 h. Fifteen minutes after induction of ischemia a significant elevation of lactate to 917 +/- 223%; p < 0.001 in the great cardiac vein could be registered. No significant changes in lactate levels were detected in the left atrium. Changes in glycerol and pyruvate showed similar patterns, with an increase to 722 +/- 297%; p < 0.001 and to 281 +/- 56%; p < 0.05, respectively. The outflow of aspartate aminotransferase and myoglobine in the great cardiac vein increased significantly. Early detection of metabolic substances is possible through the assessment of metabolic substances using microdialysis in the great cardiac vein.


Coexistence of sulfate reducers with the other oil bacterial groups in Diyarbakır oil fields.
The existence of sulfate-reducing bacteria (SRB) is a major concern in oil industry due to the detrimental effects of SRB in oil technology. SRB are co-habited with diverse microbial populations in oil fields. The presence of other bacterial groups in oil fields may alter SRB activity in different ways. Therefore, understanding this coexistence may provide insights into problems induced by SRB activity and possible solutions to these problems. To investigate this aspect, not only the presence and abundance of SRB but also bacterial population that coexists with SRB in sulfate-reducing enrichment cultures obtained from the Diyarbakır oil fields in southeast of Turkey was determined by using cultivation- and molecular-based approaches. The most probable number technique (MPN) was used to determine the number of sulfidogenic bacteria in the enrichments. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction-amplified 16S rRNA gene fragments was performed to examine the bacterial diversity of the enrichments. The results demonstrated that the number of sulfidogenic bacteria in the enrichments was low (<103 cells/mL). The DGGE analysis indicated that community members belonging to the Firmicutes were more abundant than those of other phyla. Members belonging to SRB mainly consisted of the genera Desulfosporosinus, Desulfovibrio, Thermodesulfovibrio, and Desulfotomaculum. Fermentative bacteria, acetogens, nitrate reducers, and sulfur reducers were also detected in the enrichments. The results of this study not only provide information regarding the diversity of the cultivable portion of the bacterial community that coexists with cultivable SRB, but they also offer insights into the interactions of bacteria in complex microbial communities that inhabit natural environments.


The structure of a beta-(1--6)-D-glucan from yeast cell walls.
By selective enzymolysis, or chemical fractionation, a minor polysaccharide component has been isolated from yeast (Saccharomyces cerevisiae) glucan. This minor component has a degree of polymerization of about 130-140, a highly branched structure, and a high proportion of beta-(1-->6)-glucosidic linkages. The molecules also contain a smaller proportion of beta-(1-->3)-glucosidic linkages that serve mainly as interchain linkages, but some may also be inter-residue linkages.


Heterospecific courtship, minority effects and niche separation between cryptic butterfly species.
Species interacting in varied ecological conditions often evolve in different directions in different local populations. The butterflies of the cryptic Leptidea complex are sympatrically distributed in different combinations across their Eurasian range. Interestingly, the same species is a habitat generalist in some regions and a habitat specialist in others, where a sibling species has the habitat generalist role. Previous studies suggest that this geographically variable niche divergence is generated by local processes in different contact zones. By varying the absolute and relative densities of Leptidea sinapis and Leptidea juvernica in large outdoor cages, we show that female mating success is unaffected by conspecific density, but strongly negatively affected by the density of the other species. Whereas 80% of the females mated when a conspecific couple was alone in a cage, less than 10% mated when the single couple shared the cage with five pairs of the other species. The heterospecific courtships can thus affect the population fitness, and for the species in the local minority, the suitability of a habitat is likely to depend on the presence or absence of the locally interacting species. If the local relative abundance of the different species depends on the colonization order, priority effects might determine the ecological roles of interacting species in this system.


PFA-100 System: A New Method for Assessment of Platelet Dysfunction.
This is a celebratory reprint of a historical paper published in STH in 1998. The original Abstract follows.The PFA-100 system is a platelet function analyzer designed to measure platelet-related primary hemostasis. The instrument uses two disposable cartridges: a collagen/epinephrine (CEPI) and a collagen/ADP (CADP) cartridge. Previous experience has shown that CEPI cartridges detect qualitative platelet defects, including acetylsalicylic acid (ASA)-induced abnormalities, while CADP cartridges detect only thrombocytopathies and not ASA use. In this seven-center trial, 206 healthy subjects and 176 persons with various platelet-related defects, including 127 ASA users, were studied. The platelet function status was determined by a platelet function test panel. Comparisons were made as to how well the defects were identified by the PFA-100 system and by platelet aggregometry. The reference intervals for both cartridges, testing the 206 healthy subjects, were similar to values described in smaller studies in the literature (mean closure time [CT] of 132 seconds for CEPI and 93 seconds for CADP). The use of different lot numbers of cartridges or duplicate versus singleton testing revealed no differences. Compared with the platelet function status, the PFA-100 system had a clinical sensitivity of 94.9% and a specificity of 88.8%. For aggregometry, a sensitivity of 94.3% and a specificity of 88.3% were obtained. These values are based on all 382 specimens. A separate analysis of sensitivity by type of platelet defect, ASA use versus congenital thrombocytopathies, revealed for the PFA-100 system a 94.5% sensitivity in identifying ASA users and a 95.9% sensitivity in identifying the other defects. For aggregometry, the values were 100% for ASA users and 79.6% for congenital defects. Analysis of concordance between the PFA-100 system and aggregometry revealed no difference in clinical sensitivity and specificity between the systems (p > 0.9999). The overall agreement was 87.5%, with a Kappa index of 0.751. The two tests are thus equivalent in their ability to identify normal and abnormal platelet defects. Testing 126 subjects who took 325 mg ASA revealed that the PFA-100 system (CEPI) was able to detect 71.7% of ASA-induced defects with a positive predictive value of 97.8%. The overall clinical accuracy of the system, calculated from the area under the receiver operating characteristic curve, was 0.977. The data suggest that the PFA-100 system is highly accurate in discriminating normal from abnormal platelet function. The ease of operation of the instrument makes it a useful tool to use in screening patients for platelet-related hemostasis defects.


Air density correction in ionization dosimetry.
Air density must be taken into account when ionization dosimetry is performed with unsealed ionization chambers. The German dosimetry protocol DIN 6800-2 states an air density correction factor for which current barometric pressure and temperature and their reference values must be known. It also states that differences between air density and the attendant reference value, as well as changes in ionization chamber sensitivity, can be determined using a radioactive check source. Both methods have advantages and drawbacks which the paper discusses in detail. Barometric pressure at a given height above sea level can be determined by using a suitable barometer, or data downloaded from airport or weather service internet sites. The main focus of the paper is to show how barometric data from measurement or from the internet are correctly processed. Therefore the paper also provides all the requisite equations and terminological explanations. Computed and measured barometric pressure readings are compared, and long-term experience with air density correction factors obtained using both methods is described.


Understanding oral health help-seeking among Middle Eastern refugees and asylum seekers in Australia: An exploratory study.
Andersen's behavioural model of healthcare utilization is one of the most commonly used frameworks for help-seeking across a broad range of health areas, including oral health. The Model has been used in a wide range of previous studies to explore and understand oral health service use among both general populations and ethnic minorities. However, no research has explored its utility in understanding refugees' and asylum seekers' oral health help-seeking. As such, this study aimed to improve understanding of Middle Eastern refugees' and asylum seekers' oral health help-seeking and to determine the utility of Andersen's Model in this context. Interviews were conducted with 26 participants: 20 refugees and asylum seekers, and six oral health practitioners. Results were analysed using thematic analysis. The results indicated that systemic barriers were of particular concern for refugees and asylum seekers in relation to oral healthcare access. Other components supported previous research concerning the domains of the Model, with additions in relation to acculturation and previous experiences of dental care either in home or resettlement contexts. Overall, this study highlights the need for a tailored approach to understanding oral health help-seeking for refugees and asylum seekers, who have particular dental care needs. Resettlement policies should consider system-level barriers to accessing dental services when considering access to health services more generally for refugees and asylum seekers.


Benefits of Positive Psychology-Based Story Reading on Adolescent Mental Health and Well-Being.
The importance of complete mental health in school context has recently begun to attract a lot of attention. Positive psychology interventions are often associated with improvement in mental health outcomes, but few studies have examined whether story reading is an intervention that is linked with indicators of complete mental health. This study investigated the effects of story reading interventions on both positive and negative indicators of mental health over time for a group of Turkish grade 10 high school students (n = 53). These included 33 students in a story reading group and 20 in a control group for comparison. The results showed that story reading led to improvement in students' mindfulness, optimism, happiness, and positive emotions, and also caused reduction in depression, anxiety, pessimism, and other negative emotions over a 5-week period, with a small to large effect sizes. The obtained results are discussed in the context of their implications for potential psychological interventions in high school settings.


KLOTHO allele status and the risk of early-onset occult coronary artery disease.
We previously identified a functional variant of KLOTHO (termed "KL-VS"), which harbors two amino acid substitutions in complete linkage disequilibrium and is associated with reduced human longevity when in homozygosity. Klotho-deficient mice display extensive arteriosclerosis when fed a normal diet, suggesting a potent genetic predisposition. To determine whether klotho influences atherosclerotic risk in humans, we performed cross-sectional studies to assess the association between the KL-VS allele and occult coronary artery disease (CAD) in two independent samples of apparently healthy siblings of individuals with early-onset (age <60 years) CAD (SIBS-I [N=520] and SIBS-II [N=436]). Occult CAD was defined as the occurrence of a reversible perfusion defect during exercise thallium scintigraphy and/or as an abnormal result of an exercise electrocardiogram (SIBS-I, n=97; SIBS-II, n=56). In SIBS-I, the KL-VS allele conferred a relative odds of 1.90 (95% confidence interval 1.21-2.98) for occult CAD, after adjusting for familial intraclass correlations (P<.005). Logistic regression modeling, incorporating known CAD risk factors, demonstrated that the KL-VS allele is an independent risk factor (P<.019) and that the imposed risk of KL-VS allele status is influenced by modifiable risk factors. Hypertension (P<.022) and increasing high-density lipoprotein cholesterol (HDL-C) levels (P<.022) mask or reduce the risk conferred by the KL-VS allele, respectively, whereas current smoking (P<.004) increases the risk. Remarkably concordant effects of the KL-VS allele and modifying factors on the risk of occult CAD were seen in SIBS-II. These results demonstrate that the KL-VS allele is an independent risk factor for occult CAD in two independent high-risk samples. Modifiable risk factors, including hypertension, smoking status, and HDL-C level, appear to influence the risk imposed by this allele.


Aspalathin improves hyperglycemia and glucose intolerance in obese diabetic ob/ob mice.
Although several researches have demonstrated that rooibos extract has hypoglycemic effect, the role of aspalathin, a main polyphenol in the extract, remains unclear. Our aims were to find specific mechanisms for anti-diabetic action of aspalathin employing a rat skeletal muscle-derived cell line (L6 myocytes) and a rat-derived pancreatic β-cell line (RIN-5F cells) and to investigate its effect in type 2 diabetic model ob/ob mice. We investigated in vitro the effect of aspalathin on the glucose metabolism through the studies on molecular mechanisms of glucose uptake using cultured L6 myotubes. We also measured the antioxidative ability of aspalathin against reactive oxygen species (ROS) generated by artificial advanced glycation end product (AGE) in RIN-5F cells. In vivo, ob/ob mice were fed 0.1 % aspalathin-containing diet for 5 weeks, and the effect of aspalathin on fasting blood glucose level, glucose intolerance, and hepatic gene expression was studied. Aspalathin dose dependently increased glucose uptake by L6 myotubes and promoted AMP-activated protein kinase (AMPK) phosphorylation. Aspalathin enhanced GLUT4 translocation to plasma membrane in L6 myoblasts and myotubes. In RIN-5F cells, aspalathin suppressed AGE-induced rises in ROS. In vivo, aspalathin significantly suppressed the increase in fasting blood glucose levels and improved glucose intolerance. Furthermore, aspalathin decreased expression of hepatic genes related to gluconeogenesis and lipogenesis. Hypoglycemic effect of aspalathin is related to increased GLUT4 translocation to plasma membrane via AMPK activation. In addition, aspalathin reduces the gene expression of hepatic enzymes related to glucose production and lipogenesis. These results strongly suggest that aspalathin has anti-diabetic potential.


A machine learning approach to quantify the specificity of colour-emotion associations and their cultural differences.
The link between colour and emotion and its possible similarity across cultures are questions that have not been fully resolved. Online, 711 participants from China, Germany, Greece and the UK associated 12 colour terms with 20 discrete emotion terms in their native languages. We propose a machine learning approach to quantify (a) the consistency and specificity of colour-emotion associations and (b) the degree to which they are country-specific, on the basis of the accuracy of a statistical classifier in (a) decoding the colour term evaluated on a given trial from the 20 ratings of colour-emotion associations and (b) predicting the country of origin from the 240 individual colour-emotion associations, respectively. The classifier accuracies were significantly above chance level, demonstrating that emotion associations are to some extent colour-specific and that colour-emotion associations are to some extent country-specific. A second measure of country-specificity, the in-group advantage of the colour-decoding accuracy, was detectable but relatively small (6.1%), indicating that colour-emotion associations are both universal and culture-specific. Our results show that machine learning is a promising tool when analysing complex datasets from emotion research.


Cardiopulmonary exercise testing and its application.
Cardiopulmonary exercise testing (CPET) has become an important clinical tool to evaluate exercise capacity and predict outcome in patients with heart failure and other cardiac conditions. It provides assessment of the integrative exercise responses involving the pulmonary, cardiovascular and skeletal muscle systems, which are not adequately reflected through the measurement of individual organ system function. CPET is being used increasingly in a wide spectrum of clinical applications for evaluation of undiagnosed exercise intolerance and for objective determination of functional capacity and impairment. This review focuses on the exercise physiology and physiological basis for functional exercise testing and discusses the methodology, indications, contraindications and interpretation of CPET in normal people and in patients with heart failure.


Omega-3 Polyunsaturated Fatty Acid Levels in Maternal and Cord Plasma Are Associated with Maternal Socioeconomic Status.
Omega-3 (n-3) polyunsaturated fatty acids (PUFAs) play a crucial role in fetal growth and neurodevelopment, while omega-6 (n-6) PUFAs have been associated with adverse pregnancy outcomes. Previous studies have demonstrated that socioeconomic status (SES) influences dietary intake of n-3 and n-6 PUFAs, but few studies have evaluated the association between maternal and cord plasma biomarkers of PUFAs and socioeconomic markers. An IRB-approved study enrolled mother-infant pairs (n = 55) at the time of delivery. Maternal and cord plasma PUFA concentrations were analyzed using gas chromatography. Markers of SES were obtained from validated surveys and maternal medical records. Mann-Whitney U tests and linear regression models were utilized for statistical analysis. Maternal eicosapentaenoic acid (EPA) (p = 0.02), cord EPA (p = 0.04), and total cord n-3 PUFA concentrations (p = 0.04) were significantly higher in college-educated mothers vs. mothers with less than a college education after adjustment for relevant confounders. Insurance type and household income were not significantly associated with n-3 or n-6 PUFA plasma concentrations after adjustment. Our findings suggest that mothers with lower educational status may be at risk of lower plasma concentrations of n-3 PUFAs at delivery, which could confer increased susceptibility to adverse pregnancy and birth outcomes.


Assessment of matrix Gla protein, Klotho gene polymorphisms, and oxidative stress in chronic kidney disease.
Increased vascular calcification and oxidative stress are considered as extra renal risk factors at the pathogenesis cardiovascular events in chronic kidney disease (CKD). We investigated matrix Gla protein (MGP) (T-138C, Glu60X, Thr83Ala) and Klotho (Cys370Ser) gene polymorphisms, serum MGP levels, and oxidative stress status of 84 CKD patients and 37 healthy controls. The MGP gene Glu60X and Thr83Ala polymorphisms were significantly associated with CKD. The correlation between T-138C genotype of MGP gene, Cys370Ser genotype of Klotho gene, and CKD was not significant (p > 0.05). At the haplotype analysis, the combination of the X allele of Glu60X and the Thr allele of Thr83Ala showed a significantly increased risk of CKD (p < 0.05). X allele, Thr allele, and C allele of T-138C were associated with diabetes mellitus and CKD phenotypes occurring concurrently (p < 0.01). Serum zinc levels were significantly low in end-stage renal disease (ESRD) patients (p = 0.0001). The total comet score frequency of ESRD patients was higher than that of control group (p < 0.05). The urinary 8-hydroxy-2'-deoxyguanosine levels were significantly high in CKD patients (p < 0.05). According to this study, analyzing the distribution of MGP gene and oxidative stress status would be very informative in order to detect their role at CKD.


Detection and object-based classification of offshore oil slicks using ENVISAT-ASAR images.
The aim of this study is to propose and test a multi-level methodology for detection of oil slicks in ENVISAT Advanced Synthetic Aperture Radar (ASAR) imagery, which can be used to support the identification of hydrocarbon seeps. We selected Andrusov Ridge in the Central Black Sea as the test study area where extensive hydrocarbon seepages were known to occur continuously. Hydrocarbon seepage from tectonic or stratigraphic origin at the sea floor causes oily gas plumes to rise up to the sea surface and form thin oil films called oil slicks. Microwave sensors like synthetic aperture radar (SAR) are very suitable for ocean remote sensing as they measure the backscattered radiation from the surface and show the roughness of the terrain. Oil slicks dampen the sea waves creating dark patches in the SAR image. The proposed and applied methodology includes three levels: visual interpretation, image filtering and object-based oil spill detection. Level I, after data preparation with visual interpretation, includes dark spots identification and subsets/scenes creation. After this process, the procedure continues with categorization of subsets/scenes into three cases based on contrast difference of dark spots to the surroundings. In level II, by image and morphological filtering, it includes preparation of subsets/scenes for segmentation. Level III includes segmentation and feature extraction which is followed by object-based classification. The object-based classification is applied with the fuzzy membership functions defined by extracted features of ASAR subsets/scenes, where the parameters of the detection algorithms are tuned specifically for each case group. As a result, oil slicks are discriminated from look-alikes with an overall classification accuracy of 83% for oil slicks and 77% for look-alikes obtained by averaging three different cases.


Right ventricular scar-related ventricular tachycardia in nonischemic cardiomyopathy: Electrophysiological characteristics, mapping, and ablation of underlying heart disease.
Right ventricular (RV)-scar related ventricular tachycardia (VT) is often due to arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D) or cardiac sarcoidosis (CS), but some patients whose clinical course has not been described do not fulfill diagnostic criteria for these diseases. We sought to characterize the electrophysiologic substrate and catheter ablation outcomes of such patients, termed RV cardiomyopathy of unknown source (RCUS). Data of 100 consecutive patients who presented with RV cardiomyopathy and/or RV-related VT for ablation were reviewed (51 ARVC/D, 22 CS; 27 RCUS). Compared to ARVC/D, RCUS patients were older (P = 0.001), less commonly had RV dilatation (P = 0.001) or dysfunction (P = 0.01) and fragmented QRS, parietal block, and T-wave inversion. Compared to CS, R-CUS patients had less severe LV dysfunction. Extent and distribution of endocardial/epicardial scar and inducible VTs in RCUS patients were comparable with ARVC/D and CS patients. At a median follow-up of 23 months, RCUS patients had more favorable VT-free survival (RCUS 71%, ARVC/D 60%, CS 41%, P = 0.03) and survival free of death or cardiac transplant (RCUS 92%, ARVC/D 92%, CS 62%, P = 0.01). No RCUS patients developed new criteria for ARVC/D or CS in follow-up. Up to one-third of patients with RV scar-related VT are not classifiable as ARVC/D or CS. These patients had a somewhat better prognosis than ARVC/D or sarcoid and did not develop evidence of these diseases during the initial 2 years of follow-up. The extent to which this population comprises mild ARVC/D, CS, or other diseases is not clear.


Renal sonography in the intensive care unit: when is it necessary?
To evaluate the efficacy of renal sonography performed in intensive care units on patients with the diagnosis of acute or acute-on-chronic renal failure. We reviewed all renal sonograms performed in our institution during 1 year on critically ill patients for evaluation of renal failure. Renal failure was defined as a serum creatinine level greater than 1.5 mg/dL or an increase of greater than 20% from the baseline creatinine level. Exclusion criteria included patient age younger than 18 years and signs or symptoms of obstructive uropathy. Using the electronic medical record, we recorded patient age, sex, blood urea nitrogen level, serum creatinine level, blood urea nitrogen-creatinine ratio, and clinical indication for intensive care unit admission. Sonographic reports were reviewed for the presence or absence of hydronephrosis. The total cost of these examinations was estimated with the use of Medicare reimbursement rates for 2000. One hundred five renal sonographic examinations were performed on 104 patients meeting all inclusion criteria. Only 1 study had positive results for hydronephrosis, which was graded as mild. Incidental findings not immediately affecting patient care and including ascites and simple renal cysts were identified in 91 patients. The estimated total cost of the examinations was $13,350.75. In critically ill patients with acute renal failure and no physical findings suggesting obstructive uropathy, renal sonography to evaluate for hydronephrosis is probably not indicated. This holds true regardless of patient age, sex, medical or surgical disposition, and blood urea nitrogen-creatinine ratio.


Expansion of distinct peripheral blood myeloid cell subpopulations in patients with rheumatoid arthritis-associated interstitial lung disease.
Interstitial lung disease (ILD) is associated with significant mortality in rheumatoid arthritis (RA) patients with key cellular players remaining largely unknown. This study aimed to characterize inflammatory and myeloid derived suppressor cell (MDSC) subpopulations in RA-ILD as compared to RA, idiopathic pulmonary fibrosis (IPF) without autoimmunity, and controls. Peripheral blood was collected from patients with RA, RA-ILD, IPF, and controls (N = 60, 15/cohort). Myeloid cell subpopulations were identified phenotypically by flow cytometry using the following markers:CD45,CD3,CD19,CD56,CD11b,HLA-DR,CD14,CD16,CD15,CD125,CD33. Functionality of subsets were identified with intracellular arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) expression. There was increased intermediate (CD14++CD16+) and nonclassical (CD14+/-CD16++) and decreased classical (CD14++CD16-) monocytes in RA, RA-ILD, and IPF vs. control. Intermediate monocytes were higher and classical monocytes were lower in RA-ILD vs. RA but not IPF. Monocytic (m)MDSCs were higher in RA-ILD vs. control and RA but not IPF. Granulocytic (g)MDSCs did not significantly differ. In contrast, neutrophils were increased in IPF and RA-ILD patients with elevated expression of Arg-1 sharing similar dimensional clustering pattern. Eosinophils were increased in RA-ILD vs. controls, RA and IPF. Across cohorts, iNOS was decreased in intermediate/nonclassical monocytes but increased in mMDSCs vs. classical monocytes. In RA-ILD, iNOS positive mMDSCs were increased versus classic monocytes. Myeloid cell subpopulations are significantly modulated in RA-ILD patients with expansion of CD16+ monocytes, mMDSCs, and neutrophils, a phenotypic profile more aligned with IPF than other RA patients. Eosinophil expansion was unique to RA-ILD, potentially facilitating disease pathogenesis and providing a future therapeutic target.


Minimization of Ohmic losses for domain wall motion in a ferromagnetic nanowire.
We study current-induced domain-wall motion in a narrow ferromagnetic wire. We propose a way to move domain walls with a resonant time-dependent current which dramatically decreases the Ohmic losses in the wire and allows driving of the domain wall with higher speed without burning the wire. For any domain-wall velocity we find the time dependence of the current needed to minimize the Ohmic losses. Below a critical domain-wall velocity specified by the parameters of the wire the minimal Ohmic losses are achieved by dc current. Furthermore, we identify the wire parameters for which the losses reduction from its dc value is the most dramatic.


[Transcription factors responsible for megakaryocyte-specific gene expression].
Megakaryocytes are the hematopoietic precursors of platelets, which play an essential role in thrombosis and hemostasis. Platelet factor 4 (PF4) is expressed exclusively in megakaryocytes and platelets and serves as a lineage-specific marker of megakaryocytic differentiation. We previously characterized a number of upstream enhancer and repressor elements and demonstrated that GATA-1 and ETS-1 are important for PF4 gene expression. Recently, we have determined the novel regulatory element termed "TME" in the PF4 promoter and identified a group of binding proteins from megakaryocytic HEL cells. Here we review the function of these proteins in PF4 gene expression and discuss megakaryocyte-specific gene expression and megakaryocytepoiesis.


Relation of ventricular size and function to heart failure status and ventricular dysrhythmia in patients with severe left ventricular dysfunction.
Patients with severe left ventricular (LV) dysfunction may or may not have overt heart failure and ventricular dysrhythmia. To study factors behind this variability, we examined a subset of 311 patients from the Studies of Left Ventricular Dysfunction-95 with a history of moderate heart failure (treatment trial) and 216 with no failure (prevention trial), all with ejection fractions <0.35. Echocardiographic variables were compared between trials and also correlated with dysrhythmia in 258 patients, and with neurohormones in 199 patients. Compared with prevention patients, treatment patients had larger LV end-diastolic diameter, end-systolic volume, sphericity index, and ratio of early to late diastolic filling velocity by Doppler (E/A ratio), lower LV ejection fraction and atrial contribution to ventricular filling, and similar LV mass, end-diastolic volume, and estimates of systolic wall stress. With prevention and treatment patients combined, the prevalence of abnormally elevated atrial natriuretic peptide was 92% in the highest tertile of E/A ratio compared with 55% in the lower tertiles (p=0.006). Across tertiles of LV end-diastolic volume, there was an increase in the prevalence of nonsustained ventricular tachycardia (24%, 45%, and 45%; p=0.007) and premature ventricular complexes >10/hour (48%, 62%, and 80%; p<0.001). Thus, in severe LV dysfunction, ventricular filling indexes suggestive of high filling pressures, along with larger and more spherical ventricles, are particularly common in patients with overt heart failure, thus suggesting that diastolic properties and the degree of ventricular remodeling affect clinical status. Once ejection fraction is significantly reduced, the prevalence of ventricular dysrhythmia correlates with LV size rather than systolic function. This observation lends support to previous experimental findings on the role of myocardial stretch and scar in the genesis of dysrhythmia.


PAMP-DAMPs interactions mediates development and progression of multiple sclerosis.
Multiple sclerosis (MS) is a disease presumably associated with chronic immune stimulation promoted by either pathogens or autoimmune processes. It has been hypothesized that MS could be the result of previous viral infections rendering a permanent immune stimulation that could be triggered by molecular similarities, or by modulating the antigens expression of major histocompatibility complex (MHC) on target cells, which in turn act as super antigens. During immune stimulation occurs the recruitment of immunological cells, resulting in local tissue damage and leading to the release of damage- associated molecular patterns (DAMPs), which also act as inflammation inducers. Recently, it has been proposed that the association between pathogen-associated molecular patterns (PAMPs) with DAMPs constitutes an additional level of immune regulation. The properties of DAMPs to act as carriers of PAMPs and their role as enhancers or inhibitors of PAMPs could play a role during inflammatory responses triggered by infections. Here, we focused this review in outcomes which support the hypothesis that particular PAMP-DAMPs interactions could regulated the relapse and progressive disability observed in multiple sclerosis.


Parent and physician attitudes regarding electronic communication in pediatric practices.
To determine 1) the electronic mail (e-mail) capabilities of families, general pediatricians (GPs), and subspecialty pediatricians (SPs) from an integrated pediatric health care delivery system and 2) the knowledge base and attitudes of these groups regarding the potential issues involved in using e-mail for physician-patient communication. Parents were interviewed in the offices of participating practices using a standardized survey tool. Pediatricians and staff were interviewed using a separate instrument. The data were entered into a database for analysis. A total of 325 parents and 37 physicians were interviewed. Fifty-seven percent of the 161 parents who were interviewed at the GP offices and 66% of the 164 families that were interviewed at SP offices had access to e-mail. Parents aged 31 to 40 years were significantly more likely to have access to e-mail than parents of other age groups. Access to e-mail increased with family income and parental education. Most (74%) parents who were interviewed expressed interest in using e-mail to contact their child's physician/physician's office for several purposes, including getting information or test results, scheduling appointments, and/or discussing a particular symptom. Although both groups of parents expressed concerns about confidentiality, parents at the GP offices were significantly more concerned (median(GP) = 95 vs median(SP) = 70). Seventy-four percent of GPs and 100% of SPs had access to e-mail; however, 79% did not want to use e-mail for physician-patient communication, citing concerns about confidentiality and the time demands that patient e-mail might engender. The majority of parents and pediatricians at both general and subspecialty pediatric offices are capable of communicating electronically. Parents and pediatricians are aware of the issues surrounding e-mail use for patient communication. Most parents express an interest in using e-mail for patient-physician communications, whereas most physicians are opposed to this practice.


An increased extrasynaptic NMDA tone inhibits A-type K+ current and increases excitability of hypothalamic neurosecretory neurons in hypertensive rats.
A functional coupling between extrasynaptic NMDA receptors (eNMDARs) and the A-type K+ current (IA ) influences homeostatic firing responses of magnocellular neurosecretory cells (MNCs) to a physiological challenge. However, whether an altered eNMDAR-IA coupling also contributes to exacerbated MNC activity and neurohumoral activation during disease states is unknown. We show that activation of eNMDARs by exogenously applied NMDA inhibited IA in MNCs obtained from sham, but not in MNCs from renovascular hypertensive (RVH) rats. Neither the magnitude of the exogenously evoked NMDA current nor the expression of NMDAR subunits were altered in RVH rats. Conversely, we found that a larger endogenous glutamate tone, which was not due to blunted glutamate transport activity, led to the sustained activation of eNMDARs that tonically inhibited IA , contributing in turn to higher firing activity in RVH rats. Our studies show that exacerbated activation of eNMDARs by endogenous glutamate contributes to tonic inhibition of IA and enhanced MNC excitability in RVH rats. We recently showed that a functional coupling between extrasynaptic NMDA receptors (eNMDARs) and the A-type K+ current (IA ) influences the firing activity of hypothalamic magnocellular neurosecretory neurons (MNCs), as well as homeostatic adaptive responses to a physiological challenge. Here, we aimed to determine whether changes in the eNMDAR-IA coupling also contributed to exacerbated MNC activity during disease states. We used a combination of patch-clamp electrophysiology and real-time PCR in MNCs in sham and renovascular hypertensive (RVH) rats. Activation of eNMDARs by exogenously applied NMDA inhibited IA in sham rats, but this effect was largely blunted in RVH rats. The blunted response was not due to changes in eNMDAR expression and/or function, since neither NMDA current magnitude or reversal potential, nor the levels of NR1-NR2A-D subunit expression were altered in RVH rats. Conversely, we found a larger endogenous glutamate tone, resulting in the sustained activation of eNMDARs that tonically inhibited IA and contributed also to higher ongoing firing activity in RVH rats. The enhanced endogenous glutamate tone in RVH rats was not due to blunted glutamate transporter activity. Rather, a higher transporter activity was observed, which possibly acted as a compensatory mechanism in the face of the elevated endogenous tone. In summary, our studies indicate that an elevated endogenous glutamate tone results in an exacerbated activation of eNMDARs, which in turn contributes to diminished IA magnitude and increased firing activity of MNCs from hypertensive rats.


Changes in volatile composition and sensory attributes of wines during alcohol content reduction.
A desirable sensory profile is a major consumer driver for wine acceptability and should be considered during the production of reduced-alcohol wines. Although various viticultural practices and microbiological approaches show promising results, separation technologies such as membrane filtration, in particular reverse osmosis and evaporative perstraction, in addition to vacuum distillation, represent the most common commercial methods used to produce reduced-alcohol wine. However, ethanol removal from wine can result in a significant loss of volatile compounds such as esters (ethyl octanoate, ethyl acetate, isoamyl acetate) that contribute positively to the overall perceived aroma. These losses can potentially reduce the acceptability of the wine to consumers and decrease their willingness to purchase wines that have had their alcohol level reduced. The change in aroma as a result of the ethanol removal processes is influenced by a number of factors: the type of alcohol reduction process; the chemical-physical properties (volatility, hydrophobicity, steric hindrance) of the aroma compounds; the retention properties of the wine non-volatile matrix; and the ethanol level. This review identifies and summarises possible deleterious influences of the dealcoholisation process and describes best practice strategies to maintain the original wine composition. © 2016 Society of Chemical Industry.


O6-methylguanine DNA methyltransferase status determined by promoter methylation and immunohistochemistry in gliosarcoma and their clinical implications.
O(6)-methylguanine-DNA methyltransferase (MGMT) is known as a DNA repair protein, and loss of function in MGMT is related to an increase in survival in patients with malignant gliomas treated with alkylating agents. In the present study, we determined the status of MGMT using methylation-specific polymerase chain reaction (PCR) and immunohistochemistry on paraffin-embedded specimens in 12 human gliosarcomas, and these results were then related to overall survival (OS) and response to alkylating agents. The MGMT promoter was methylated in six patients. Immunostaining of MGMT was positive in 58.3% of patients. MGMT methylation status was correlated with immunostaining results in five patients (41.7%). The median OS and progression-free survival (PFS) of the whole population were 13.4 months [95% confidence interval (CI), 12.3-14.5 months] and 8.3 months (95% CI, 7.4-9.2 months), respectively. In patients with methylated MGMT promoter, median OS was 15.0 months, compared with 11.3 months in the unmethylated group. Median PFS of gliosarcoma patients was 10.3 months for the methylated group, whereas it was 7.3 months for the unmethylated group. On multivariate analysis, patients with methylated MGMT promoter had better prognosis than patients with unmethylated MGMT promoter with respect to OS and PFS (P = 0.045 and 0.034, respectively). However, there was no statistical significance between MGMT protein expression and survival. The results show that a significant fraction of gliosarcomas have MGMT promoter methylation and protein expression, and suggest that patient survival is associated with MGMT methylation status.


Serum creatinine vs. albuminuria as biomarkers for the estimation of cardiovascular risk.
Cardiovascular (CV) disease is the leading cause of morbidity and mortality among people with chronic kidney disease (CKD). CKD has increased over the past decade to become a worldwide public health problem. The definition of a biomarker is a characteristic objectively measured and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacologic response to a therapeutic intervention. Thus, biomarkers of kidney function would include serum creatinine and more recently estimated glomerular filtration rate (eGFR). These biomarkers and microalbuminuria, potential biomarker, predict CV events and mortality. Recent analyses of cross-sectional data indicate that eGFR is a much stronger predictor of CV events than is microalbuminuria. While microalbuminuria indicates endothelial dysfunction and is associated with increased risk for CV events, its level is related more to the level of blood pressure and glycemic control than directly to the pathophysiology of atherosclerosis. Hence, microalbuminuria could be viewed as a biomarker but not a risk factor for CV risk since, risk factors must be an integral part of the disease pathophysiology. Conversely, while microalbuminuria is not of prognostic value to predict CKD outcomes, increases over time into the albuminuria range, >200 mg/day, clearly indicate presence of kidney disease and are associated with a more rapid decline in kidney function. Thus, concomitant evaluation of both biomarkers eGFR and albuminuria is recommended to assess kidney function and CV risk thoroughly.


Transcription of histone gene cluster by differential core-promoter factors.
The 100 copies of tandemly arrayed Drosophila linker (H1) and core (H2A/B and H3/H4) histone gene cluster are coordinately regulated during the cell cycle. However, the molecular mechanisms that must allow differential transcription of linker versus core histones prevalent during development remain elusive. Here, we used fluorescence imaging, biochemistry, and genetics to show that TBP (TATA-box-binding protein)-related factor 2 (TRF2) selectively regulates the TATA-less Histone H1 gene promoter, while TBP/TFIID targets core histone transcription. Importantly, TRF2-depleted polytene chromosomes display severe chromosomal structural defects. This selective usage of TRF2 and TBP provides a novel mechanism to differentially direct transcription within the histone cluster. Moreover, genome-wide chromatin immunoprecipitation (ChIP)-on-chip analyses coupled with RNA interference (RNAi)-mediated functional studies revealed that TRF2 targets several classes of TATA-less promoters of >1000 genes including those driving transcription of essential chromatin organization and protein synthesis genes. Our studies establish that TRF2 promoter recognition complexes play a significantly more central role in governing metazoan transcription than previously appreciated.


Experimental arthritis induced by atypical strains of Streptococcus pyogenes.
Experimental arthritis was induced in Sprague-Dawley rats by intraarticular injection of whole-cell sonicates, heat-killed cells and cell-wall preparations of typical and atypical strains of Group-A streptococci (Streptococcus pyogenes). The non-haemolytic nitrosoguanidine-derived mutant and the naturally occurring Lowry strain induced a similar but less severe form of arthritis. Direct immunofluorescent staining demonstrated maximum fluorescence in the sections of articular joint taken 60 days after injection. The level of immune complexes increased for up to 90 days after injection of cell walls or whole-cell sonicates and correlated well with the development of the chronic stage of arthritis observed in haematoxylin and eosin and fluorescence staining of thin tissue sections.


Rate constants and kinetic isotope effects for H-atom abstraction reactions by muonium in the Mu + propane and Mu + n-butane reactions from 300 K to 435 K: challenges for theory.
This paper reports measurements of the temperature dependence of the rate constants for H-atom abstraction reactions from propane and n-butane by the light isotopic H-atom muonium (Mu), kMu(T), over temperatures in the range 300 K to 435 K. Simple Arrhenius fits to these data yield activation energies, E, that are some 2-4 times lower than E found from corresponding fits for the H + propane and H + n-butane reactions studied elsewhere, both experimentally and theoretically, and fit over a similar temperature range. These activation energies E are also much lower than estimated from zero-point-energy corrected vibrationally adiabatic potential barriers, both results suggesting that quantum tunneling plays an important role in determining kMu(T) and for the Mu + propane reaction in particular. The results are expected to pose a considerable challenge to reaction rate theory for isotopic H-atom reactions in alkane systems.


Venlafaxine extended release vs placebo and paroxetine in social anxiety disorder.
Evidence indicates that venlafaxine hydrochloride extended release (ER) effectively ameliorates anxiety symptoms. To evaluate the efficacy, safety, and tolerability of flexible-dose venlafaxine ER compared with placebo in the short-term treatment of generalized social anxiety disorder and, secondarily, to compare paroxetine with venlafaxine ER and paroxetine with placebo. Adult outpatients with DSM-IV generalized social anxiety disorder for 6 months or longer were randomly assigned to receive venlafaxine hydrochloride ER (75-225 mg/d), paroxetine (20-50 mg/d), or placebo for 12 weeks or less at 26 centers in the United States. The primary outcome measure was the total Liebowitz Social Anxiety Scale score. Secondary measures included response (Clinical Global Impression-Improvement score, 1 or 2) rates and Clinical Global Impression-Severity of Illness and Social Phobia Inventory scores. Of 440 patients treated, 413 (93.9%) were included in the last-observation-carried-forward efficacy analysis; of the 429 patients in the safety population, 318 (74.1%) completed the study. Mean daily doses were 201.7 mg (SD, 38.1 mg) of venlafaxine hydrochloride ER and 46.0 mg (SD, 7.9 mg) of paroxetine. Venlafaxine ER treatment was significantly superior to placebo at weeks 1 through 12 on the Liebowitz Social Anxiety Scale and Social Phobia Inventory and at week 2 and weeks 6 through 12 for Clinical Global Impression-Severity of Illness and responder status, and was significantly superior to paroxetine treatment at weeks 1 and 2 for the Social Phobia Inventory (P < .05 for all). Paroxetine treatment was significantly superior to placebo at weeks 3 through 12 on the Liebowitz Social Anxiety Scale, the Clinical Global Impression-Severity of Illness scale, and the Social Phobia Inventory, and at weeks 4 through 12 for response (P < .05 for all). Week 12 response rates were significantly greater for the venlafaxine ER and paroxetine groups (58.6% and 62.5%, respectively) vs the placebo group (36.1%) (P < .001 for both). Venlafaxine ER is effective in the short-term treatment of generalized social anxiety disorder, with efficacy and tolerability comparable to paroxetine.


Determinants of antiretroviral therapy adherence among older adolescents living with HIV in Kenya during the transition to adult care; An observational study.
Lower levels of adherence to antiretroviral therapy (ART) among older adolescents as compared to adults are influenced by individual, psychosocial, and treatment-related factors. Successful transition of older adolescents into HIV adult care from paediatric & adolescent focused care requires an understanding of barriers to ART adherence. This study aimed at determining individual factors affecting ART adherence among older HIV positive adolescents transitioning to adult care. Between December 2018 and January 2019, we conducted a cross-sectional study among 82 perinatally infected adolescents aged 16-19 years in an HIV care and treatment clinic in Nairobi, Kenya. We used completed structured questionnaires and abstracted data from clinical charts. We performed multivariate logistic regression to identify factors independently associated >95% self-reported ART adherence (7-day recall). The study participants had a median age of 17 (IQR 16,18) on ART for a median duration of 11 years (IQR 7,13). Sixty-four per cent (52) of the adolescents reported optimal adherence was of >95%, and 15% reported missing doses for three or more months. Self -reported adherence had a high correlation with viral loads of <1000 copies ml (Kappa= 0.087). Adolescents with high self-efficacy were eight times more likely to report adherence of >95% [OR 8.1, 95% CI (2.31- 28.18)]. Once a day, dosing was also independently associated with adherence [OR 1.58, 95 %CI [0.62-4.08]. The reduction of ART pill burden and the inclusion of assessment of ART self -efficacy may contribute to transition preparedness among adolescents.


Beam deflection measurement of time and polarization resolved ultrafast nonlinear refraction.
We modify the well-known photothermal beam deflection technique to study ultrafast nonlinearities. Using phase-sensitive detection we directly measure the temporal and polarization dynamics of nonlinear refraction (NLR) with sensitivity to optically induced phase changes of approximately λ/20,000. We use the relative polarization dependence of excitation and probe to separate the isotropic and reorientational components of the NLR.


Intraluminal pressure oscillation enhances subsequent airway contraction in isolated bronchial segments.
A period of deep inspiration in humans has been shown to attenuate subsequent bronchoconstriction, a phenomenon termed bronchoprotection. The bronchoprotective effect of deep inspiration may be caused though a depression in the force production of airway smooth muscle (ASM). We determined the response of whole airway segments and isolated ASM to a period of cyclic stretches. Isovolumetric contraction to electrical field stimulation (EFS) was assessed in porcine bronchial segments before and after intraluminal pressure oscillation from 5 to 25 cmH(2)O for 10 min at 0.5 Hz. Morphometry showed that this pressure oscillation stretched ASM length by 21%. After pressure oscillation, the response to EFS was not reduced but instead was modestly enhanced (P < 0.01). Airway responses to EFS returned to preoscillation levels 10 min after the end of oscillation. The increase in EFS response after pressure oscillation was not altered by the addition of indomethacin. In a separate experiment, we assessed isometric force in isolated ASM strips before and after length oscillation. The amplitude, frequency, and duration of length oscillation were similar to those induced in bronchial segments. In contrast to bronchial segments, length oscillation of ASM produced a significant depression in isometric force induced by EFS (P < 0.01). These results suggest that the response of ASM to length oscillation is modified by the airway wall. They also suggest that the phenomenon of bronchoprotection reported in some in vivo studies may not be an intrinsic property of the airway.


Contribution of BKCa channels to vascular tone regulation by PDE3 and PDE4 is lost in heart failure.
Regulation of vascular tone by 3',5'-cyclic adenosine monophosphate (cAMP) involves many effectors including the large conductance, Ca2+-activated, K+ (BKCa) channels. In arteries, cAMP is mainly hydrolyzed by type 3 and 4 phosphodiesterases (PDE3, PDE4). Here, we examined the specific contribution of BKCa channels to tone regulation by these PDEs in rat coronary arteries, and how this is altered in heart failure (HF). Concomitant application of PDE3 (cilostamide) and PDE4 (Ro-20-1724) inhibitors increased BKCa unitary channel activity in isolated myocytes from rat coronary arteries. Myography was conducted in isolated, U46619-contracted coronary arteries. Cilostamide (Cil) or Ro-20-1724 induced a vasorelaxation that was greatly reduced by iberiotoxin (IBTX), a BKCa channel blocker. Ro-20-1724 and Cil potentiated the relaxation induced by the β-adrenergic agonist isoprenaline (ISO) or the adenylyl cyclase activator L-858051 (L85). IBTX abolished the effect of PDE inhibitors on ISO but did not on L85. In coronary arteries from rats with HF induced by aortic stenosis, contractility and response to acetylcholine were dramatically reduced compared with arteries from sham rats, but relaxation to PDE inhibitors was retained. Interestingly, however, IBTX had no effect on Ro-20-1724- and Cil-induced vasorelaxations in HF. Expression of the BKCa channel α-subunit, of a 98 kDa PDE3A and of a 80 kDa PDE4D were lower in HF compared with sham coronary arteries, while that of a 70 kDa PDE4B was increased. Proximity ligation assays demonstrated that PDE3 and PDE4 were localized in the vicinity of the channel. BKCa channels mediate the relaxation of coronary artery induced by PDE3 and PDE4 inhibition. This is achieved by co-localization of both PDEs with BKCa channels, enabling tight control of cAMP available for channel opening. Contribution of the channel is prominent at rest and on β-adrenergic stimulation. This coupling is lost in HF.


Gp130 signaling promotes development of acute experimental colitis by facilitating early neutrophil/macrophage recruitment and activation.
IL-6 is known to play a crucial role in the pathogenesis of chronic intestinal inflammation by modulating T cell functions. In this study, we investigated the role of gp130, the common signal transducer for all IL-6 cytokines, in a murine model of acute T cell independent colitis to better characterize the impact of gp130 on innate immune cells and the early stages of inflammation. Experimental colitis was induced by dextran sulfate sodium treatment of mice with inducible systemic deletion of gp130 (MxCre/gp130(-/-)), macrophage/neutrophil-specific gp130-deficiency (LysCre/gp130(-/-)), or bone marrow chimeric mice and compared with wild-type controls (gp130(f/f)). Systemic deletion of gp130 (MxCre/gp130(-/-)) protected mice from severe colitis and wasting and attenuated the mucosal inflammatory infiltrate as well as local cytokine, chemokine, and adhesion molecule expression. Experiments in newly generated macrophage/neutrophil-specific gp130-deleted animals (LysCre/gp130(-/-)) and gp130 bone marrow chimeric mice, revealed a dual mechanism of proinflammatory effects mediated by gp130. Leukocyte recruitment was impaired in gp130-deleted animals and gp130-deleted recipients of wild-type bone marrow, demonstrating a central role of gp130-dependent signals in nonmyeloid cells for directing leukocytes to sites of inflammation, which was further confirmed in a model of sterile peritonitis. In contrast, macrophage/neutrophil-specific gp130 deficiency delayed and attenuated the disease but only marginally affected the inflammatory infiltrate, indicating a defective activation of mucosal leukocytes. We provide evidence that IL-6 cytokines acting via gp130 are required in the acute stages of intestinal inflammation by modulating the dynamics of innate immune cell recruitment and activation.


Improved release of triamcinolone acetonide from medicated soft contact lenses loaded with drug nanosuspensions.
Drug nanosuspensions (NSs) show a significant potential to improve loading and release properties of the poorly water soluble drug triamcinolone acetonide (TA) from poly(hydroxyethyl methacrylate) (pHEMA) soft contact lenses. In this work, TA NSs were developed by a controlled precipitation method using a fractional factorial Plackett-Burmann design. Poloxamer 407 (PL) and polyvinyl alcohol (PVA) as stabilizing agents were selected. NSs were characterized in terms of their drug content, particle size and morphology. Results indicate that all studied factors, except homogenization speed and sonication, have significant influence on the drug incorporation yield into NSs. Drug nanoparticles showed an interesting size that may be suitable for their incorporation into topical ocular drug delivery systems, as hydrogels. pHEMA hydrogels and daily-wear Hilafilcon B commercial contact lenses (SCLs) were employed to study TA loading capacity and drug release properties using NSs as loading system. Hydrogels have been synthesised by copolymerization of 2-hydroxyethyl methacrylate (HEMA) with methacrylic acid (MA) in accordance with a previous work (García-Millán et al., 2015). Both synthesised hydrogels and SCLs were characterized in terms of their mechanical and physical properties and TA loading and release properties. Selected TA NS was further characterized by studying its physical-chemical stability during the loading process. Results show that the use of TA NSs as loading medium significantly increases drug loading capacity and release of soft contact lenses in comparison with drug saturated solution. Synthesised pHEMA hydrogels and SCLs lenses have good properties as ophthalmic drug delivery systems, but SCLs load higher quantities of drug and release TA in shorter time periods than synthesised pHEMA hydrogel.


Antidiabetes Agents and Incident Depression: A Nationwide Population-Based Study.
Diabetes is associated with an increased risk of depression. Some antidiabetes agents, specifically metformin and pioglitazone, have been suggested to have beneficial effects on depression, but associations between antidiabetes drugs and depression have not been systematically investigated. We combined four Danish population-based registers to investigate whether the 20 most widely used orally administered antidiabetes drugs were associated with an altered risk of incident depression. Analyses of insulin were included for comparisons. All persons in Denmark in 2005 were included in the study and followed until 2015. Two different outcome measures of incident depression were included: 1) a diagnosis of depressive disorder at a psychiatric hospital as an inpatient or outpatient and 2) a combined measure of a diagnosis of depression or use of antidepressants. Data were analyzed using Cox regression models. A total of 360,205 individuals using orally administered antidiabetes drugs and 64,582 using insulin at any time during the study period were included in the analyses. Continued use of metformin and combinations of drugs including metformin were associated with decreased rates of incident depression. Pioglitazone was not associated with a decreased rate of incident depression. No other antidiabetes drugs or insulin showed significant associations with depression. Real-life population-based data suggest a positive effect of metformin on depression rates. This evidence should be used in guiding prescriptions for patients with type 2 diabetes who are at risk for developing depression, including those with prior depression or anxiety and patients with a family history of depression.


Differential activation of dorsal basal ganglia during externally and self paced sequences of arm movements.
The basal ganglia are thought to be critically involved in motor control. However, the relative contributions of the various sub-components are not known. Although, in principle, functional magnetic resonance imaging (fMRI) provides adequate resolution to image the basal ganglia at the spatial scale of the individual nuclei, activating these nuclei with fMRI has proven to be difficult. Here we report two tasks, involving externally and self paced sequences of arm movements, which resulted in significant activation of contralateral posterior (post-commissural) putamen and globus pallidus. This activation did not significantly differ between the tasks. In contrast, significant activation of the contralateral and ipsilateral anterior caudate and anterior putamen was observed only during externally paced arm movements. These results suggest a dissociation in the roles of the anterior and posterior dorsal basal ganglia: the anterior caudate and putamen may be involved in sensory to motor mapping and the posterior putamen and globus pallidus may be involved in the motor response itself. The findings support the hypothesis that the basal ganglia may be involved in gating sensory influences onto motor areas.


Exploring ward nurses' perceptions of continuing education in clinical settings.
Health care systems demand that nurses are flexible skilful workers who maintain currency and competency in order to deliver safe effective patient centered care. Nurses must continually build best practice into their care and acquire lifelong learning. Often this learning is acquired within the work environment and is facilitated by the clinical nurse educator. Understanding clinical nurses' values and needs of continuing education is necessary to ensure appropriate education service delivery and thus enhance patient care. To explore clinical ward-based nurses' values and perceptions towards continuing education and what factors impact on continuing education in the ward. A case study approach was utilized. A major teaching hospital in Melbourne, Australia. A range of clinical nursing staff (n=23). Four focus groups and six semi-structured individual interviews were undertaken. Focus group interviews explored participants' values and perceptions on continuing education through a values clarification tool. Thematic analysis of interviews was undertaken to identify themes and cluster data. Three central themes: 'culture and attitudes', 'what is learning?' and 'being there-being seen', emerged reflecting staffs' values and perceptions of education and learning in the workplace. Multiple factors influence ward nurses' ability and motivation to incorporate lifelong learning into their practice. Despite variance in nurses' values and perceptions of CE in clinical environments, CE was perceived as important. Nurses yearned for changes to facilitate lifelong learning and cultivate a learning culture. Clinical nurse educators need to be cognizant of adult learners' characteristics such as values, beliefs, needs and potential barriers, to effectively facilitate support in a challenging and complex learning environment. Organizational support is essential so ward managers in conjunction with educational departments can promote and sustain continuing education, lifelong learning and a culture conducive to learning.


Pathophysiology of ischemic cell death: III. Role of extracellular factors.
The purpose of this study was to determine the effect on cell survival of extracellular changes that occur during ischemia, over and above the depletion of O2 and substrate. Rabbit retinas were deprived in vitro of both O2 and substrate, and then returned to control medium for 4 h before recovery was assessed by measuring protein synthesis, glucose utilization, and tissue water. Experimental conditions were altered in various ways during the period of O2 and substrate deprivation in order to modify the changes taking place in the interstitial fluid as a result of the failure of energy metabolism. When O2-free, substrate-free extracellular electrolyte solution was added to the retinas to reduce the ischemia-induced changes in the interstitial fluid, there was marked reduction in irreversible damage. But when energy-deprived retinas were exposed to retinas that had already been ischemic, or to interstitial fluid from ischemic retinas, there was an increase in irreversible damage. Removing Ca++ from the extracellular fluid during the period of energy deprivation increased the damage due to short deprivations in a restricted volume of extracellular fluid, but reduced the damage from longer deprivations in a large volume of extracellular fluid. The results demonstrate that several changes occur in the extracellular fluid during ischemia that significantly affect recovery.


Relevance of a portable spirometer for detection of small airways obstruction.
While portable spirometers are increasingly used, little attention has been paid to test their validity for measurement of flows in small airways. The aim of this study was to compare the Spirotel portable spirometer to a laboratory spirometer (Jeager PFT), with regard to accuracy in measuring forced expiratory flows, and more specifically those influenced by small airways (FEF(25-75)). Fifty-nine children (mean age, 12 years; range, 7-17), were studied at baseline and after a bronchodilator inhalation. Spirometers were tested separately in a randomly designed order. A total of 117 sessions of flow-volume curves was performed with each spirometer. We obtained at least two acceptable and reproducible curves in 88% and 76% of the sessions, with the laboratory and the portable spirometers, respectively. Unacceptable curves were easily detected by visual inspection of flow-time and flow-volume waveforms. Agreement was excellent between spirometers for the measurement of all expiratory flows, both at baseline and postbronchodilator. More specifically, agreement between spirometers was as high for measurements of FEF(25-75) (intraclass correlation coefficients 0.97) as for proximal flows. High correlations were found between baseline expiratory flows measured by each spirometer (and expressed as percent of predicted values), both in large and small airways (P < 0.001). The portable spirometer was highly sensitive for detecting small airways obstruction, as compared to the laboratory spirometer. Finally, the magnitudes of bronchodilator-related flow changes were also highly correlated, both in large and small airways (P < 0.001 and P = 0.004, respectively). We conclude that the Spirotel portable spirometer is reliable for measurement of forced expiratory flows, in large and small airways, provided that all curve waveforms can be stored and available for visual inspection.


Reverse causal reasoning: applying qualitative causal knowledge to the interpretation of high-throughput data.
Gene expression profiling and other genome-scale measurement technologies provide comprehensive information about molecular changes resulting from a chemical or genetic perturbation, or disease state. A critical challenge is the development of methods to interpret these large-scale data sets to identify specific biological mechanisms that can provide experimentally verifiable hypotheses and lead to the understanding of disease and drug action. We present a detailed description of Reverse Causal Reasoning (RCR), a reverse engineering methodology to infer mechanistic hypotheses from molecular profiling data. This methodology requires prior knowledge in the form of small networks that causally link a key upstream controller node representing a biological mechanism to downstream measurable quantities. These small directed networks are generated from a knowledge base of literature-curated qualitative biological cause-and-effect relationships expressed as a network. The small mechanism networks are evaluated as hypotheses to explain observed differential measurements. We provide a simple implementation of this methodology, Whistle, specifically geared towards the analysis of gene expression data and using prior knowledge expressed in Biological Expression Language (BEL). We present the Whistle analyses for three transcriptomic data sets using a publically available knowledge base. The mechanisms inferred by Whistle are consistent with the expected biology for each data set. Reverse Causal Reasoning yields mechanistic insights to the interpretation of gene expression profiling data that are distinct from and complementary to the results of analyses using ontology or pathway gene sets. This reverse engineering algorithm provides an evidence-driven approach to the development of models of disease, drug action, and drug toxicity.


Staged management of open Lisfranc injury: Experience from 14 patients.
There are still controversies on the management and outcome of open Lisfranc injury in available studies. This study evaluates the staged management of Lisfranc injury and its complications.Patients who received a staged strategy for open Lisfranc injury were reviewed.One patient with degloving injury suffered from partial skin and hallux necrosis which was treated by debridement, hallux amputation, definitive internal fixation, and local flap transfer on the 12th day after first stage management. A definitive internal fixation and simultaneous skin graft or flap coverage were performed in another 3 patients with soft tissue defects. Other patients without soft tissue problems underwent a second stage of definitive internal fixation. Bone union was observed on the 12.5th week after definitive surgery. The median AOFAS midfoot score at the last follow-up was 74.4 ± 8.7, while the average VAS score was 2.2 ± 1.8. The average return-to-work time was 8th months postoperatively in 9 patients. Flap necrosis, infection, implant failure, nonunion, and osteomyelitis were not observed during the follow-up. Two patients received Lisfranc arthrodesis for persistent pain due to posttraumatic arthritis.In the management of open Lisfranc injury, surgeons must consider soft tissue condition. Staged strategy is a rational protocol for this severe injury. Temporary K-wire fixation after early radical debridement and realignment will facilitate the definitive internal fixation until soft tissue condition improves, which also can decrease the soft tissue complication.


Age-associated disruption of molecular clock expression in skeletal muscle of the spontaneously hypertensive rat.
It is well known that spontaneously hypertensive rats (SHR) develop muscle pathologies with hypertension and heart failure, though the mechanism remains poorly understood. Woon et al. (2007) linked the circadian clock gene Bmal1 to hypertension and metabolic dysfunction in the SHR. Building on these findings, we compared the expression pattern of several core-clock genes in the gastrocnemius muscle of aged SHR (80 weeks; overt heart failure) compared to aged-matched control WKY strain. Heart failure was associated with marked effects on the expression of Bmal1, Clock and Rora in addition to several non-circadian genes important in regulating skeletal muscle phenotype including Mck, Ttn and Mef2c. We next performed circadian time-course collections at a young age (8 weeks; pre-hypertensive) and adult age (22 weeks; hypertensive) to determine if clock gene expression was disrupted in gastrocnemius, heart and liver tissues prior to or after the rats became hypertensive. We found that hypertensive/hypertrophic SHR showed a dampening of peak Bmal1 and Rev-erb expression in the liver, and the clock-controlled gene Pgc1α in the gastrocnemius. In addition, the core-clock gene Clock and the muscle-specific, clock-controlled gene Myod1, no longer maintained a circadian pattern of expression in gastrocnemius from the hypertensive SHR. These findings provide a framework to suggest a mechanism whereby chronic heart failure leads to skeletal muscle pathologies; prolonged dysregulation of the molecular clock in skeletal muscle results in altered Clock, Pgc1α and Myod1 expression which in turn leads to the mis-regulation of target genes important for mechanical and metabolic function of skeletal muscle.


EFFECTS OF BICARBONATE ON GROWTH OF PASTEURELLA PESTIS. I. DIFFERENTIAL RESPONSE OF VIRULENT AND AVIRULENT CELLS.
Surgalla, M. J. (Fort Detrick, Frederick, Md.), A. W. Andrews, and C. L. Baugh. Effects of bicarbonate on growth of Pasteurella pestis. I. Differential response of virulent and avirulent cells. J. Bacteriol. 88:269-272. 1964.-Virulent Pasteurella pestis and the avirulent mutants always present in a virulent inoculum have been demonstrated to give growth responses in opposite directions upon addition of bicarbonate to broth cultures under certain conditions. The effect of supplemental bicarbonate on initiation of growth from a virulent inoculum was found to be either stimulation of both virulent and avirulent cells at low levels of added bicarbonate, depression of both virulent and avirulent cells at higher initial concentrations, or depression of avirulent mutants with simultaneous stimulation of virulent cells at a narrow range of intermediate levels.


Regulation of proteins mediating neurodegeneration in experimental autoimmune encephalomyelitis and multiple sclerosis.
Multiple sclerosis is characterized by inflammatory demyelination and axonal loss as pathophysiological correlates of relapsing activity and progressive development of clinical disability. The molecular processes involved in this pathogenesis are still unclear as they are quite complex and heterogeneous. In this article we present protein expression analysis of brain and spinal cord tissues from different models of murine experimental autoimmune encephalomyelitis (EAE), the most commonly used animal model for multiple sclerosis. We observed a number of EAE-specific protein expression and PTM differences. Proteome analysis was extended to multiple sclerosis specimens in order to validate the EAE findings. Our findings suggest the regulation of a number of proteins that shed light on the molecular mechanisms of the disease processes taking place in EAE and multiple sclerosis. We found consistent modulation of proteins including serum amyloid P component, sirtuin 2, dihydropyrimidinase-related protein family proteins, stathmin 1, creatine kinase B and chloride intracellular channel protein 1. Functional classification of the proteins by database and the literature mining reveals their association with neuronal development and myelinogenesis, suggesting possible disease processes that mediate neurodegeneration.


Variation in site of death among nursing home residents in British Columbia, Canada.
To assess the proportion of in-hospital versus in-nursing home deaths among a population of decedent nursing home residents in British Columbia, Canada, and to identify facility and individual characteristics associated with in-hospital death. We examined nursing home (ownership/organization, size) and individual (age, level of care, sex, previous hospitalization within 30 days) characteristics of all decedent residents of British Columbia's freestanding publicly funded nursing homes. Secondary administrative data from the Ministry of Health, supplemented with facility data were analyzed. The study population included those aged 65 years and older who died between April 1, 1996 and August 1, 1999 (n = 14,413). Mixed models were used to estimate unadjusted and adjusted odds ratios (AOR; 95% confidence intervals [CI]) for factors associated with in-hospital death. Almost one quarter (24.6%) of deaths occurred in hospital. In-hospital death was more frequent in nonprofit (NP) single-site facilities compared to NP facilities owned and/or operated by a health authority (AOR = 1.37, 95% CI: 1.15, 1.64). Smaller nursing home size (AOR = 1.25, 95% CI: 1.05, 1.50) and male gender (AOR = 1.17, 95% CI: 1.07, 1.27) were also associated with a greater odds of in-hospital death. Progressively lower odds ratios of in-hospital death were observed for each category of increasing age and declining function, respectively. While individual characteristics play a significant role in explaining variation in site of death, residence in a NP single-site and smaller-sized facility was also associated with a greater frequency of in-hospital death.


Sub-20 fs pulses shaped directly in the UV by an acousto-optic programmable dispersive filter.
Direct pulse shaping in the UV was used to compress and structure pulses throughout the range of 250 - 400 nm. Broadband pulses generated by SHG of a NOPA were used as input to an acousto-optic programmable dispersive filter. As this shaper creates lateral dispersion, aspects of Gaussian and geometric optics had to be considered for the design of the beam path. Special care was taken to produce a homogeneous input beam. We show nearly Fourier-limited pulses as short as 16.8 fs at 320 nm and 19.5 fs at 260 nm. Full control over amplitude and phase is demonstrated by generating arbitrary shapes like square pulses and complex pulse sequences. The subpulses were manipulated individually in intensity, temporal delay, chirp, relative phase and central wavelength.


Activation of ataxia telangiectasia-mutated DNA damage checkpoint signal transduction elicited by herpes simplex virus infection.
Eukaryotic cells are equipped with machinery to monitor and repair damaged DNA. Herpes simplex virus (HSV) DNA replication occurs at discrete sites in nuclei, the replication compartment, where viral replication proteins cluster and synthesize a large amount of viral DNA. In the present study, HSV infection was found to elicit a cellular DNA damage response, with activation of the ataxia-telangiectasia-mutated (ATM) signal transduction pathway, as observed by autophosphorylation of ATM and phosphorylation of multiple downstream targets including Nbs1, Chk2, and p53, while infection with a UV-inactivated virus or with a replication-defective virus did not. Activated ATM and the DNA damage sensor MRN complex composed of Mre11, Rad50, and Nbs1 were recruited and retained at sites of viral DNA replication, probably recognizing newly synthesized viral DNAs as abnormal DNA structures. These events were not observed in ATM-deficient cells, indicating ATM dependence. In Nbs1-deficient cells, HSV infection induced an ATM DNA damage response that was delayed, suggesting a functional MRN complex requirement for efficient ATM activation. However, ATM silencing had no effect on viral replication in 293T cells. Our data open up an interesting question of how the virus is able to complete its replication, although host cells activate ATM checkpoint signaling in response to the HSV infection.


Nanoscopic polypyrrole AFM-SECM probes enabling force measurements under potential control.
Conductive polymers, and in particular polypyrrole, are frequently used as biomimetic interfaces facilitating growth and/or differentiation of cells and tissues. Hence, studying forces and local interactions between such polymer interfaces and cells at the nanoscale is of particular interest. Frequently, such force interactions are not directly accessible with high spatial resolution. Consequently, we have developed nanoscopic polypyrrole electrodes, which are integrated in AFM-SECM probes. Bifunctional AFM-SECM probes were modified via ion beam-induced deposition resulting in pyramidal conductive Pt-C composite electrodes. These nanoscopic electrodes then enabled localized polypyrrole deposition, thus resulting in polymer-modified AFM probes with a well-defined geometry. Furthermore, such probes may be reversibly switched from an insulating to a conductive state. In addition, the hydrophilicity of such polymer tips is dependent on the dopant, and hence, on the oxidation state. Force studies applying different tip potentials were performed at plasma-treated glass surfaces providing localized information on the associated force interactions, which are dependent on the applied potential and the dopant.


Characterization of the prostate-specific antigen (PSA) catalytic mechanism: a pre-steady-state and steady-state study.
Prostate-specific antigen (PSA), an enzyme of 30 kDa grouped in the kallikrein family is synthesized to high levels by normal and malignant prostate epithelial cells. Therefore, it is the main biomarker currently used for early diagnosis of prostate cancer. Here, presteady-state and steady-state kinetics of the PSA-catalyzed hydrolysis of the fluorogenic substrate Mu-His-Ser-Ser-Lys-Leu-Gln-AMC (spanning from pH 6.5 to pH 9.0, at 37.0°C) are reported. Steady-state kinetics display at every pH value a peculiar feature, represented by an initial "burst" phase of the fluorescence signal before steady-state conditions are taking place. This behavior, which has been already observed in other members of the kallikrein family, suggests the occurrence of a proteolytic mechanism wherefore the acylation step is faster than the deacylation process. This feature allows to detect the acyl intermediate, where the newly formed C-terminal carboxylic acid of the cleaved substrate forms an ester bond with the -OH group of the Ser195 catalytic residue, whereas the AMC product has been already released. Therefore, the pH-dependence of the two enzymatic steps (i.e., acylation and deacylation) has been separately characterized, allowing the determination of pKa values. On this basis, possible residues are tentatively identified in PSA, which might regulate these two steps by interacting with the two portions of the substrate.