Temperature‐responsive genetic loci in pectinolytic plant pathogenic Dickeya solani

Fifty‐four Dickeya solani thermoregulated genes were identified using Tn5 transposon mutagenesis with an inducible gusA reporter system; 45 genes were up‐regulated at 37 °C, whereas nine were up‐regulated at 18 °C. The relative level of gene up‐regulation ranged from 2–1200 and 5–650 U/mg total proteins at 18 and 37 °C, respectively. Among the temperature‐regulated loci, genes coding for proteins involved in fundamental bacterial metabolism, membrane‐related proteins and pathogenicity‐corresponding factors and several hypothetical unknown proteins were found. The mutants were tested for their pathogenicity in planta and for features known to be important for D. solani virulence viz. production of pectinolytic enzymes, cellulases, proteases, siderophores and auxins as well as for motility and the ability to form a biofilm. Eight Tn5 mutants, four up‐regulated at high and four up‐regulated at low temperature, expressed visible phenotypes including the decreased ability to cause symptoms on potato tubers and chicory leaves, impairment in phospholipase production and/or deficiency in biofilm formation. The implications of environmental temperature on the ability of D. solani to cause disease symptoms in potato are discussed.     

Identification and characterization of long non-coding RNAs in porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

Background: Long non-coding RNAs(lncRNAs) may regulate gene expression in numerous biological processes including cellular response to xenobiotics.The exposure of living organisms to 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD),a persistent environmental contaminant,results in reproductive defects in many species including pigs.The aims of the study were to identify and characterize lncRNAs in porcine granulosa cells as well as to examine the effects of TCDD on the lncRNA expression profile in the cells.Results: One thousand six hundred sixty-six lncRNAs were identified and characterized in porcine granulosa cells.The identified lncRNAs were found to be shorter than mRNAs.In addition,the number of exons was lower in lncRNAs than in m RNAs and their exons were longer.TCDD affected the expression of 22 lncRNAs(differentially expressed lncRNAs [DELs]; log2 fold change ≥ 1,P-adjusted 0.05) in the examined cells.Potential functions of DELs were indirectly predicted via searching their target cis-and trans-regulated protein-coding genes.The coexpression analysis revealed that DELs may influence the expression of numerous genes,including those involved in cellular response to xenobiotics,dioxin metabolism,endoplasmic reticulum stress and cell proliferation.Aryl hydrocarbon receptor(AhR) and cytochrome P450 1 A1(CYP1 A1) were found among the trans-regulated genes.Conclusions: These findings indicate that the identified lncRNAs may constitute a part of the regulatory mechanism of TCDD action in granulosa cells.To our knowledge,this is the first study describing lncRNAs in porcine granulosa cells as well as TCDD effects on the lncRNA expression profile.These results may trigger new research directions leading to better understanding of molecular processes induced by xenobiotics in the ovary.     

Community level physiological profiles of microbial communities from forest humus polluted with different amounts of Zn,Pb, and Cd—Preliminary study with BIOLOG ecoplates

Abstract

Heavy metals may alter the structure and metabolic functions of soil microbial community. The objective of our study was to compare the community level physiological proffies (CLPPs) of microbial communities from forest humus polluted with different amounts of Zn, Pb, and Cd to test whether the addition of soluble Zn and Cd may affect the CLPPs of microbial communities. The samples were taken at 18 locations in southern Poland referred to as unpolluted (UP), slightly polluted (SP), and heavily polluted sites (HP). The contents of heavy metals were measured after wet digestions in concentrated HNO₃. Microbial communities were extracted using 0.96% NaCl solution. In order to test heavy metal tolerance of microbial communities from UP sites the extracts from these sites were additionally treated with Zn (50 mg L^-1; UP + Zn) and Cd (1 mg L^-1; UP + Cd). Metabolic functions of the microbial communities were analyzed using BIOLOG Ecoplates method. The contents of Zn, Pb and Cd were the highest at HP sites (4,740, 1,120, 41.0 mg kg^-1, respectively) followed by SP (830, 509, 9.2 mg kg^-1, respectively), and UP (173, 93, 2.1 mg kg^-1, respectively) sites. Principal components analysis (PCA) indicated that CLPPs at all sites were similar. This suggests that microbial community from SP and HP sites revealed tolerance to heavy metals. Addition of Zn affected CLPPs of microbial communities from UP sites as indicated by significantly (p < 0.05) higher value of PC1 score. The addition of Cd did not affect CLPPs of microbial communities from these sites.     

Stress Reactivity and Coping in Horse Adaptation to Environment

“Coping” or “coping strategy”, defined as the behavioral and physiological efforts of animal to master the situation, is more and more in interest of researchers dealing with applied animal behavior and welfare. Knowledge about “coping styles” may be helpful in understanding individual adaptive capacity to stressful events. At least two types of animal coping strategy (or coping style) can be involved: (1) active copers (proactive) and (2) passive copers (reactive). The individual differences in stress response to threatening situations have been found in horses, and these animals can show specific coping strategies such as other species. This article reviews the set of behavioral, psychoneuroendocrine, and psychoneuroimmune mechanisms involved in animal adaptation to environmental challenges and discusses the relationship between behavioral and physiological factors involved in stress response of the horse. Exploring coping strategies in horses and in any captive animal can be successful when a multidimensional approach including behavioral, neural, hormonal, and hematological measures is considered. Knowledge on stress coping styles can provide valuable information to predict the behavior of individuals during response to specific challenging situations. Moreover, assessing individual differences in adaptation strategies can be useful in horse selection to different exploitation tasks and reproduction.     

Growth factor and cytokine interactions in myogenesis. Part II. Expression of IGF binding proteins and protein kinases essential for myogenesis in mouse C2C12 myogenic cells exposed to TNF-alpha and IFN-gamma

The aim of the study was to examine potential interactions among IGF-I and proinflammatory cytokines, TNF-alpha and IFN-gamma, in the regulation of local IGF-I bioavailability and cellular proteins mediating myogenic signals. We investigated levels of IGFBP-4, -5, -6, protein kinase Czeta (PKC zeta), p38 and extracellular signal-regulated kinase (ERK1/2) in differentiating mouse C2C12 myoblasts. IGF-I significantly stimulated expression of IGFBP-5. TNF-alpha and IFN-gamma attenuated the expression of IGFBP-4 and -6 under basal conditions and in the presence of IGF-I, and inhibited IGF-I-induced IGFBP-5 expression during 5-day myogenesis. TNF-alpha and IFN-gamma markedly attenuated p38 expression in the presence of IGF-I on the 5th day of myogenesis. When combined with IGF-I the cytokines exerted opposite effects on the PKC zeta level, i.e. TNF-alpha caused an increase, whereas IFN-gamma reduced the cellular content of this kinase. Exposition of C2C12 myoblasts to IGF-I or cytokines led to the stimulation of ERK1/2 phosphorylation; however, both TNF-alpha and IFN-gamma exerted an inhibitory effect on the activation of ERK1/2 in myoblasts cultured in the presence of IGF-I. We concluded as follows: i) TNF-alpha and IFN-gamma present in the extracellular environment of differentiating C2C12 myoblasts can alter the local bioavailability of IGF-I by inhibiting the expression of IGFBP-4, -5, and -6, ii) the decrease in p38 expression and ERK1/2 phosphorylation in C2C12 myoblasts exposed to cytokines can lead to disturbances in IGF-I-regulated myogenesis.     

The effect of "Ferrodex", vitamin A and "Rovimix beta-carotene" on the iron and copper levels in blood plasma of calves

The effect of Ferrodex, vitamin A and synthetic beta-carotene on the content of iron and copper in the blood serum has been investigated in three groups of calves: I control group without vitamin supplements, II group received only once with the first colostrum dose 1.5 million IU of vitamin A, III group received Rovimix beta carotene containing 10 per cent of the synthetic beta carotene with Mlekomix from the 21st day to the 13th week of life. Moreover half of the calves from each group received Ferrodex intramusculary. Content of iron, total iron binding capacity by serum proteins, level of indirect and direct copper as well as the content of ceruloplasmin were determined in the blood serum of calves. Moreover hematocrit and hemoglobin content in the blood were determined. The results obtained show that a triple intramuscular injection of Ferrodex caused an increase of the level of iron, copper and ceruloplasmin in the blood serum of calves. However, a single administration of vitamin A to the calves or a continuous administration of Rovimix beta-carotene decreased significantly the level of iron and ceruloplasmin copper in blood serum as well as caused a more dynamic ceruloplasmin synthesis with a simultaneous increase in its utilization.     

Anti-oxidative/oxidative status of rat liver after ovariectomy

The lack of protective action of oestrogens which appears during menopausal period may be the reason of serious metabolic disturbances including oxidative stress. The hypothesis was stated that ovariectomy may induce the variations of antioxidant/oxidant status which can be detected in rat liver. A total of 102 healthy Wistar female rats were included in the experiment and divided into control (CON; n = 6), sham-operated (SHO; n = 48) and ovariectomized (OVX; n = 48) groups. Animals from SHO (n = 6) and OVX (n = 6) groups were killed every week during 8 weeks of experiment to detect dynamic changes in examined parameters. Anti-oxidative enzyme activities [glutathione peroxidase (GSH-Px); superoxide dismutase (SOD)] as well as total antioxidant capacity (TAC) and the intensity of lipid peroxidation, measured by the concentration of N,N,diethyl-p-phenylene diamine (DEPPD) radical, were determined in liver homogenates by the use of spectrophotometric methods. Wave-like patterns of examined parameters within 8 weeks of experiment were detected. GSH-Px activity tended to be higher in OVX animals and was significantly lower at 8th week when compared with 1st week of experiment. SOD activity was higher in SHO animals and showed significant differences between 3rd, 4th, 7th and 1st week. TAC values were significantly higher in OVX when compared with SHO groups in 2nd, 4th, 5th week and significantly lower in 3rd, 6th and 7th week of the experiment. The concentration of DEPPD radicals tended to increase in OVX group. In conclusion, ovariectomy which leads to oestrogen insufficiency is reflected as well in variation of anti-oxidative/oxidative parameters in rat liver homogenates.     

Expression of connexin 43 protein in testes, epididymides and prostates of stallions

REASONS FOR PERFORMING STUDY: Connexin 43 (Cx43) is a ubiquitously distributed gap junction protein in testes and other reproductive tissues. Adjacent cells share ions and small metabolites through intercellular channels, which are present in gap junctions. Previously, Cx43 has not been reported in testes, epididymides and prostates either in healthy stallions or cryptorchid horses. OBJECTIVES: To demonstrate the expression pattern of Cx43 in the reproductive tissues of stallions and examine whether naturally occurring bilateral cryptorchidism has any influence on distribution and expression of Cx43. METHODS: The expression and the presence of Cx43 protein were detected by means of immunohistochemistry and Western blot analysis using a polyclonal rabbit anti-Cx43 antibody. RESULTS: In stallions, gap junctions appeared as structures localised to cell-cell contacts between adjacent cells. In testes, Cx43 expression was detected in the interstitial tissue and seminiferous tubules, between Leydig and Sertoli, as well as Sertoli and germ cells. In epididymides, Cx43 was localised between epithelial cells, whereas in prostates, between secretory cells of the glandular epithelium. In the cryptorchid, a clear reduction of Cx43 signal was observed in all reproductive tissues. CONCLUSIONS: Coupling of Leydig cells via gap junctions may suggest that steroidogenic function of the testis is under the influence of these intercellular channels. Within seminiferous tubules, the expression was found to be stage-specific, pointing to its role in coordinating spermatogenesis. Differential distribution of Cx43 protein in the reproductive tract of normal and cryptorchid stallions indicates that expression is clearly dependent on the physiological status of the horse. POTENTIAL RELEVANCE: Detection of Cx43 expression in equine testicular, epididymal, and prostatic cells is important for a better understanding of the role of intercellular membrane channels in direct cell communication within the reproductive tract of stallions.