Background: Excess plugging of small airways is associated with premature death in chronic obstructive pulmonary disease (COPD). Over-expression of beta-epithelial sodium channel (β-ENaC) in airway epithelia in mice resulted in plugging of small airways while cystic fibrosis transmembrane regulator (CFTR) negatively regulated ENaC activity in cell models. Purpose: To test the hypothesis that accumulation of mucus exudates observed with the progression of COPD is related to excess airway epithelial sodium re-absorption as a result of over-expression of β-ENaC and reduced expression of CFTR by small airway epithelia. Methods: Small airway epithelial samples from frozen lungs from patients at different levels of COPD severity were isolated by laser capture microdissection (LCM). β-ENaC, CFTR, and β-actin (control) gene expression was determined by qRT-PCR and compared to expression in entire airways and lung parenchyma surrounding these airways. β-ENaC protein as well as epithelial mucin expression and mucus plugging were localized and quantified after immunohistochemical and periodic acid Schiff staining, respectively. Results: β-ENaC mRNA expression had a strong positive correlation with that of CFTR (p<O.0001) in airway epithelia and surrounding lung parenchyma (p=O.Ol) but not whole airways. β-ENaC mRNA and protein expression were positively correlated (p=O.4O, p=O.O5) and protein expression significantly increased with GOLD stage of COPD severity. Epithelial mucin expression positively correlated with β-ENaC (p=O.38, p=O.O5) and CFTR (p=OAO, p=O.O4.) mRNA and with mucus plugging (p=O. 43 , ptO.OOO2). CFTR mRNA also correlated positively with mucus plugging (p=O. 48 , p=O.O2). Conclusions: Strong positive correlations between β-ENaC and CFTR mRNA expression that are limited to the lung parenchyma and epithelium suggest a novel mechanism of mRNA regulation. This differs from their functional relationship where an inverse relationship between CFTR expression and β-ENaC activity has been reported. Positive correlations of epithelial mucin or mucus plugging with CFTR mRNA but not β-ENaC protein expression in the small airway epithelium suggest that CFTR may regulate mucin at this site independently of β-ENaC protein. The relationship between β-ENaC mRNA andepithelial mucin expression could be due to strong correlations between β-ENaC and CFTR mRNA expression but β-ENaC’s relationship with COPD GOLD stage suggests it may nevertheless play a role in COPD.

In the work of Master both self-expression of teenagers and facilities performing songs solo have been examined. It was determined that during the stage of adolescence way of performance singing solo depends on both psychological and physiological qualities which decide child’s musical potentiality of self-expression. This kind of self-expression allows the personality to unfold and realise oneself in the activity of music. Conceptions and opinions as the key teaching form of music about singing based on various authors have been revised in the work, too. However, when performing songs solo during the stage of adolescence it evokes negative emotions. First part of the work of Master discusses psychological and physiological peculiarities and conceptions of self-expression of a teenager. It has been determined that the following factors have strong influence on teenager’s personality and possibility to express oneself through the singing solo: emotions, requirements, interests, motivation and self-confidence. Second part of the work explains the point of the singing solo as well as means and factors of the impact of an educator which allow the student to show oneself actively during the course of singing solo. Various ways of solo performance have been displayed and described. The results of empirical research are stated in the third part of the work. It has been aspired to study the possibilities of self-expression of the students of 6-8th form when performing songs solo and... [to full text]

The aim of the present work has been to analyze the ways of adolescents’ anger expression and to estimate the relations between emotional and behavioral problems. There was collected data about 198 11-18 years old adolescents. In the research there were used YSR 11/18: Youth self – report (Achenbach, 1991), TRI - Interpersonal Reactivity Index (Davis, 1980) and STAXI – C/A– State - Trait Anger Expression Inventory – 2 For Children and Adolescents (Spielberger, 1999). The results of the research show that there is a positive correlation between anger expression index and internal behavioral problems (value of p is 0.00) and external behavioral problems (value of p is 0.00). Internal behavioral problems also correlate with anger as a state (value of p is 0.00), anger as a trait (value of p is 0.00), internal (value of p is 0.00) and external (value of p is 0.00) anger expression, internal anger control (value of p is 0.02). External behavioral problems correlate with anger as a state (value of p is 0.00), anger as a trait (value of p is 0.00) and external anger expression (value of p is 0.00). There was estimated a negative correlation between adolescents’ anger as a state and their competence (value of p is 0.02). A positive correlation estimated between external anger expression and social activity (value of p is 0.01). A positive correlation also estimated between internal anger control and learning (value of p is 0.03). Anger of adolescents is also related with... [to full text]

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Ornithine-$ delta$-biaminotransferase (OAT) is a mitochondrial matrix a enzyme which catalyzes the reversible conversion of ornithine to glutamic semialdehyde. In the retinoblastoma cell line Y79, the 5$ sp prime$ UTR of the OAT mRNA is alternatively-spliced. Previous work has shown that the two alternatively-spliced 5$ sp prime$ UTR are translated with different efficiencies. / The alternatively-spliced 5$ sp prime$ UTRs were isolated and subcloned. Expression vectors were constructed containing each of the OAT 5$ sp prime$ UTRs and the human growth hormone as a reporter gene. Transient transfection expression analysis in COS-1 cells confirmed that the 5$ sp prime$ UTRs conferred differential expression on the reporter gene. The shorter 5$ sp prime$ UTR was expressed at a level 3-fold higher than that of the longer 5$ sp prime$ UTR. / RNA mobility shift assays were performed to determine whether a transacting factor was interacting with one of the alternatively-spliced 5$ sp prime$ UTRs. A binding activity was detected that bound to the longer, exon 2-containing 5$ sp prime$ UTR with a significantly higher affinity than the shorter 5$ sp prime$ UTR. Competition studies demonstrated that this binding activity was specific for the longer 5$ sp prime$ UTR.

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Transport stress research has shown correlations among stress, morbidity, and mortality in calves subjected to the traditional U.S. market system, indicating the possibility of compromised immune function. The objective of this study was to determine if expression of specific immune and inflammatory response genes differed between calves that were subjected to either an acute stress (AS, handled and weaned for 1.5 h) or a chronic stress (CS, weaned, handled and transported for 3 to 4 d). Two groups of forty calves, Bos taurus (n = 20) and crossbred calves (n = 20), weighing 181 kg to 250 kg were used in each of two trials. Jugular veni-puncture blood samples (9 ml) were collected from AS calves 1.5 h after the start of handling and separation from their dam. Samples were collected from CS calves during processing after arrival at a north Texas feed lot. RNA for gene expression analysis was extracted from peripheral blood leukocytes obtained from blood samples by a filtration method. During the second trial, the filtrate was centrifuged for measurement of plasma cortisol. A diagonal covariance mixed model ANOVA was used to determine effects of treatment, breed, and breed by treatment interaction on cortisol concentrations. Expression values for each gene were analyzed using linear models that considered the effects of treatment (AS and CS) and breed (Bos taurus and crossbred calves) comparing each trial separately. Mean plasma cortisol concentrations did not differ between AS (16.40 +/- 1.08 ng/ml) and CS calves (18.06 +/- 1.14 ng/ml) (P > 0.296). The interaction of effects was detected for 2 genes in Trial 1, and 3 genes in Trial 2 (P < 0.029). Breed was influential for 5 genes in both Trial 1 and 2 (P < 0.046). Significant differences were found in relative quantification for 30 genes in Trial 1 and 36 genes in Trial 2, in which CS calves had greater expression than AS calves (P < 0.047). Fifteen of those genes were common between the two trials with mean treatment differences of RQ values from the 15 genes ranging from 0.309 to 913.19, excluding outliers. Similar elevated cortisol concentrations in both the AS and CS calves indicated that both groups experienced significant stress. However, changes in gene expression differences were greater in the calves subjected to CS, indicating that gene expression may be more useful than cortisol for identifying detrimental long-term stress.

Rhodococcus fascians, a plant pathogenic actinomycete with a very broad host range, causes leafy galls and other malformations. The plant hormone, group, the cytokinins has been implicated in the alteration of host morphology. The aim of this project was to gain insight into the interaction of the cytokinin biosynthetic, isopentenyltransferase (IPT), cytokinin activating ( LOG (The Lonely Guy)) and the cytokinin metabolic, cytokinin oxidase/dehydrogenase (CKX) gene families of both Pisum sativum and R. fascians during infection of the plant. R. fascians colonisation and infection of pea were examined using scanning electron microscopy (SEM) and light microscopy. The expression of genes related to cytokinin biosynthesis, activation and metabolism were isolated and assessed with polymerase chain reaction (PCR) and real time-quantitative PCR (RT-qPCR) analysis. Primers were designed to discriminate between pea genes and R. fascians genes. In addition, the response of the pea cotyledons to R. fascians was measured through chlorophyll estimation and the expression of the transporter genes, sucrose transporter (SUT) and amino acid permease (AAP) which were assayed through RT-qPCR. The pea response regulators were monitored as an indirect measure of the level of endogenous cytokinins in pea. Two R. fascians strains, the avirulent strain 589 and the virulent strain 602, were selected for this project based on their virulence and similar growth rate under identical conditions. The virulence of R. fascians virulent strain 602 was also confirmed through Koch's postulates. The phenotypic alterations in the pea infected with the virulent strain 602 included stunted growth, multiple shoots, small leaves, thickened primary roots and reduced secondary root growth. Delayed senescence of shoots and dark green, intact cotyledons were also observed. Microscopic analyses revealed epiphytic colonisation by both the avirulent strain 589 and the virulent strain 602 in pea cotyledons, roots, shoots and leaves and endophytic colonisation in the seed coat from the time of seed inoculation to 45 days post inoculation (dpi). The expression of R. fascians genes was relatively high at 5 and 9 dpi in pea cotyledons and at 15 and 25 dpi in roots and shoots of pea infected with the virulent strain 602. The expression of RfIPT, RfLOG and RfCKX was not detected both in the control pea and the pea infected with the avirulent strain 589. The cytokinin biosynthesis, metabolism and response regulator (RR) multi-gene families of PsIPTs, PsLOGs, PsCKXs and PsRRs revealed differential and tissue-specific expression patterns. The expression of PsIPTs and PsLOGs was induced immediately after inoculation with the R. fascians virulent strain 602 in the cotyledon but not in roots and shoots, and the expression level reduced at later growth stages. The PsCKXs and PsRRs expression level increased with the growth of the host infected with the virulent strain 602. In pea infected with the avirulent strain 589 the expression of PsIPTs, PsLOGs and PsCKXs gene family members generally increased after 25 dpi in cotyledons, roots and shoots, whereas PsRRs expression was low at all time points. The up-regulation of PsIPTs and PsLOGs immediately after inoculation in cotyledons and at 15 dpi in roots and shoots by R. fascians virulent strain leads to elevated cytokinins which is reflected by the up-regulation of PsRRs. The plant responds to elevated cytokinin by producing phenotypic changes including shoot malformations. The plant activates its cytokinin homeostasis mechanism due to change in cytokinin level which is indicated by up-regulation of PsCKXs. Generally, the expression of the PsRRs was also up-regulated over time following infection by the R. fascians virulent strain. This indicates the presence of biologically active cytokinins in the host which maintain the symptoms. The outcome due to the avirulent strains indicates that, even though PsIPTs and PsLOGs are up-regulated at later growth stages (25 to 35 dpi), expression of PsCKX gene families were varied (either up-regulated or down regulated after 25 dpi). However, PsRRs expression was down-regulated suggesting low cytokinins levels in tissues which may be due to the activation of homeostatic mechanisms of the plant to reduce the level of biologically active cytokinins. The chlorophyll content increased in cotyledons infected with the virulent strain 602 and PsSUTs and PsAAPs expression pattern in pea cotyledon and shoot infected with the virulent strain 602 indicates that R. fascians converts the infected tissue into a sink for their establishment and growth.