Effect of conjugated linoleic acids on the activity and mRNA expression of 5- and 15-lipoxygenases in human macrophages

Lipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies on conjugated linoleic acids (CLAs) as fatty acids with a potential anti-atherosclerotic function have recently been initiated. The aim of the study was to test the effect of CLAs and linoleic acid on 5- and 15-lipoxygenase (5-LO, 15-LO-1) enzyme activity, their mRNA expression, and concentration in the cells. It was also desired to determine whether the CLAs are substrates for the enzymes. For the experiments monocytic cell line (THP-1) and monocytes obtained from human venous blood were used. Monocytes were differentiated to macrophages: THP-1 (CD14+) by PMA administration (100 nM for 24 h) and monocytes from blood (CD14+) by 7-day cultivation with the autologous serum (10%). After differentiation, macrophages were cultured with 30 microM CLAs or linoleic acid for 48 h. The 15- and 5-lipoxygenase products were measured by HPLC method. mRNA expression and protein content were analyzed by real-time PCR and Western blot analysis. The in vitro studies proved that both CLA isomers are not substrates for 15-LO-1; in ex vivo studies hydroxydecadienoic acid (HODE) concentration was significantly reduced (p = 0.019). The trans-10,cis-12 CLA isomer reduced HODE concentration by 28% (p = 0.046) and the cis-9,trans-11 CLA isomer by 35% (p = 0.028). In macrophages obtained from THP-1 fatty acids did not change significantly mRNA expression of the majority of the investigated genes. CLAs did not change the content of 5-LO and 15-LO-1 proteins in macrophages obtained from peripheral blood. Linoleic acid induced 15-LO-1 expression (2.6 times, p < 0.05). CLAs may perform the function of an inhibitor of lipoxygenase 15-LO-1 activity in macrophages.     

Genetic data from avian influenza and avian paramyxoviruses generated by the European network of excellence (EPIZONE) between 2006 and 2011--review and recommendations for surveillance

Since 2006, the members of the molecular epidemiological working group of the European "EPIZONE" network of excellence have been generating sequence data on avian influenza and avian paramyxoviruses from both European and African sources in an attempt to more fully understand the circulation and impact of these viruses. This review presents a timely update on the epidemiological situation of these viruses based on sequence data generated during the lifetime of this project in addition to data produced by other groups during the same period. Based on this information and putting it all into a European context, recommendations for continued surveillance of these important viruses within Europe are presented.     

2,4-D and MCPA and their derivatives: Effect on the activity of membrane erythrocytes acetylcholinesterase (in vitro)

The effect of 2,4-dichlorophenoxyacetic acid (2,4-D), 4-chloro-2-methylphenoxyacetic acid (MCPA), and their derivatives: phenol, 2,4-dichlorophenol (2,4-DCP), 2,4-dimethylphenol (2,4-DMP), and catechol on the activity of acetylcholinesterase (AChE, EC 3.1.1.7) in human erythrocytes was studied. Phenol, MCPA, and 2,4-DMP did not significantly change AChE activity in human erythrocytes (in vitro). Decrease of AChE activity was observed under the highest applied dose of 2,4-D—500 and 1000 ppm. Decrease of AChE activity exposed to 2,4-DCP and catechol was noted and depended on the doses of applied compounds. The relationship between activities and substrate concentrations (curves) was analyzed for reactions of acetylcholinesterase. Catalytic constants K_m and V_max were calculated from the Michaelis curve. Statistically significant decrease of V_max and K_m was observed in the activity of AChE incubated with 2,4-DCP and catechol, revealing mixed inhibition type of AChE inhibition (this compound may affect not only on enzyme but also on complex ES as well). 2,4-D decreases V_max but do not change K_m value, what reveals non-competitive type of AChE inhibition by this compounds. Non-competitive inhibition does not depend on the substrate concentrations but only on the inhibitor concentration and its K_i value, characterizes the affinity of inhibitors towards enzyme. In conclusion, changes of AChE activity upon 2,4-D, 2,4-DCP, and catechol are the consequences of direct interactions between compounds and the enzyme and indirect via membrane modification and increase of Reactive Oxygen Species.     

Seroprevalence of Neospora caninum in dogs in south-western Poland

The prevalence of anti-Neospora caninum antibodies was determined in serum samples from 110 dogs in south-western Poland, using the indirect fluorescent antibody test (IFAT). Antibodies to N. caninum were found in 18 dogs (16.36%), of which 7 were also positive to Toxoplasma gondii. The titres of anti-N. caninum antibodies varied from 1:50 to 1:200.     

Phylogenetic background, virulence gene profiles, and genomic diversity in commensal Escherichia coli isolated from ten mammal species living in one zoo

Three hundred commensal Escherichia coli recovered from healthy herbivorous, carnivorous, and omnivorous mammals from one zoo were characterized for their phylogenetic origin, intestinal virulence gene (VG) prevalence, and genomic diversity. The phylogenetic structure of the E. coli (groups A, B1, B2, and D) from the herbivores was homogenous, with a prevailing representation of group B1. In the carnivores and omnivores, the phylogenetic diversity was species specific with a higher representation of group A compared to the herbivores. Of 16 intestinal VGs in the whole set, 8 were detected and they formed 13 VG profiles. In the herbivores, all the VG-positive isolates belonged to group B1 and harboured the genes eaeA, eastI, ehxA, stx1, and stx2, which separately or in combination formed 8 VG profiles. In the carnivores and omnivores, the VG-positive isolates frequently belonged to group A and harboured the estI and estII genes or a combination of eastI and estI, forming three VG profiles. Single genes cnf2, in group B2, and eastI, in group D, were found. Similarity analysis of pulsed-field gel electrophoresis (PFGE) patterns revealed closer relatedness between the isolates from carnivores and omnivores than those from herbivores. The comparison between the prevalence of phylogenetic groups and the phylogenetic origin of VG-positive isolates in the examined E. coli suggested, that E. coli from group B1 in herbivores and E. coli from group A rather than B1 in carnivores and omnivores are "best adapted" to the host organism. The groups revealed different preferences in the acquisition and maintenance of intestinal VGs.     

The role of floods and droughts on riverine ecosystems under a changing climate

Floods and droughts are key driving forces shaping aquatic ecosystems. Climate change may alter key attributes of these events and consequently health and distribution of aquatic species. Improved knowledge of biological responses to different types of floods and droughts in rivers should allow the better prediction of the ecological consequences of climate change‐induced flow alterations. This review highlights that in unmodified ecosystems, the intensity and direction of biological impacts of floods and droughts vary, but the overall consequence is an increase in biological diversity and ecosystem health. To predict the impact of climate change, metrics that allow the quantitative linking of physical disturbance attributes to the directions and intensities of biological impacts are needed. The link between habitat change and the character of biological response is provided by the frequency of occurrence of the river wave characteristic—that is the event's predictability. The severity of impacts of floods is largely related to the river wave amplitude (flood magnitude), while the impact of droughts is related to river wavelength (drought duration).     

Dispersal and mortality of translocated thick‐shelled river mussel Unio crassus Philipsson, 1788 adults revealed by radio tracking

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The effects of heavy metals on embryonic development of fish (a review)

Early developmental stages of fish are particularly sensitive to water pollution. Heavy metals may affect various developmental processes during the embryonic period, which results in a reduction of offspring quantity and quality. Waterborne metals may accumulate in the gonads of spawners and adversely affect gamete production and viability, or exert direct toxic influence upon developing embryos. The egg shell does not fully protect the embryo against metal penetration, particularly during the swelling phase; thus, metals may accumulate in the egg. The results depend on metal concentration and range from developmental disturbances to death of the embryo. Metals disturb various processes of fish embryonic development and affect the development rate. Early stages just after fertilization are particularly sensitive to metal intoxication, when most disturbances and the highest embryonic mortality occur. Waterborne metals also promote developmental anomalies during organogenesis, including body malformations. Heavy metals often induce a delay in the hatching process, premature hatching, deformations and death of newly hatched larvae. All these disturbances result in reduced numbers and poor quality of the larvae, which show small body size, high frequency of malformations and reduced viability.     

Dependence of the lactation duration and efficiency on the season of lambing in relation to the prolactin and melatonin secretion in ewes

Changes in day length and the related secretion of melatonin and prolactin are of particular significance in sheep, as they determine reproductive processes, the last stage of which is lactation. The aim of the present study was to examine the dependence of the lactation duration and efficiency on the season of lambing (January—Group I/June—Group II) in relation to the hormonal status of milking ewes (Polish Longwool). The possibility of artificial extension of the milking period in late-lambing ewes by application of prolonged day length, 16 h of light–8 h of darkness (16L:8D), was introduced additionally (Group III). Measurements of plasma levels of prolactin and melatonin were used as parameters of season-dependent hormonal regulation of milk production in this seasonally breeding species. Although during the first 28 days of lactation sheep of all groups produced similar amounts of milk (based on the weight gains of the lambs), our results clearly showed that during the period of machine milking (from weaning at 56 day to the dry period) the level of milk production in Group I was almost twice that noted in Group II. Thus, the highest milk yield was obtained from sheep during the period when the days became longer, i.e. from March to May, which was accompanied by an increase in prolactin secretion. In both groups, melatonin secretion was found to increase while milk performance parameters and prolactin secretion decreased during the shortening photoperiod. The sustained decreases in milk production and prolactin secretion were also observed during the autumn months in sheep from Group III, despite the low level of melatonin. The present study showed that time of lambing and related photoperiod length could significantly affect the milk production of sheep that have a clear season of sexual activity. Maintenance of June-lambing sheep under the 16L:8D regime failed to extend the high prolactin level and lactation considerably into the autumn months. The spontaneous decrease in prolactin secretion observed under the long artificial photoperiod indicated for a development of refractoriness, which could also limit the lactation period.     

Effect of melatonin on biochemical variables of the blood in dairy cows

In order to examine the effect of exogenous melatonin on selected biochemical variables of the blood in ruminants, dairy cows were given the pineal gland hormone in the dose of 0.1 mg/kg body weight. One and four hours after melatonin administration blood samples were collected from the cows in the control and the treated group in order to determine the levels of glucose, insulin, total cholesterol, triglycerides, free fatty acids, as well as the activities of alanine and aspartate aminotransferase. The pineal gland hormone caused a significant increase in the levels of total cholesterol and triglycerides, slight increases in glucose and insulin levels, and a significant decrease in the concentration of free fatty acids. Melatonin did not exert an effect on the activity of liver enzymes.