Effects of prostaglandin F(2alpha) and nitric oxide on the secretory function of bovine luteal cells

The objective of the present study was to investigate the influence of prostaglandin F(2alpha) (PGF (2alpha)) and nitric oxide (NO) on production of steroids and PGs by culturing bovine luteal cells obtained from ovaries on days 8-12 of the estrous cycle with a nitric oxide (NO) donor (Spermine NONOate), and a NO synthase inhibitor (N(G)-nitro-L-arginine methyl ester dihydrochloride: L-NAME). When the cells were exposed for 24 h to PGF(2alpha) (10(-7)-10(-5) M), production of progesterone (P(4)) increased significantly at all doses used (P<0.05). Moreover, PGF(2alpha) stimulated PGF(2alpha) production (P<0.01), depressed testosterone (T) production (P<0.05), but did not affect synthesis of prostaglandin E(2) (PGE(2)). Spermine NONOate decreased P(4) production to 66%, 47% and 34% of the control concentration after treatment with 10(-5) M, 10(-4) M and 10(-3) M, respectively, but did not affect T production, and increased PGF(2alpha) synthesis (P<0.05) and PGE(2) (P<0.01) at all doses used. L-NAME increased production of P(4) (P<0.01) but did not affect (P>0.05) secretion of T, PGF(2alpha) and PGE(2). Estradiol-17beta (E(2)) was detectable on the level of sensitivity of assay and was not significantly altered by any treatments. The overall results suggest that PGF(2alpha) and NO produced locally in bovine CL play roles in the regulation of the secretory function of the bovine CL as auto/paracrine factors.     

Identification of bap and icaA genes involved in biofilm formation in coagulase negative staphylococci isolated from feline conjunctiva

Bap and icaA genes are commonly known to be involved in the biofilm formation. The prevalence of bap and icaA genes and biofilm formation was determined in conjunctival isolates of coagulase negative staphylococci (CNS) collected from cats. The study was conducted on 90 archival CNS isolates collected from feline conjunctiva obtained from clinically healthy cats and cats with ocular problems. Biofilm formation was examined using the microtiter plate (MTP) method. The prevalence of icaA and bap genes was determined using polymerase chain reaction (PCR). Genetic profiles of the bap-positive isolates were examined using the modified random amplified polymorphic DNA (RAPD) method. Of the 90 CNS isolates investigated, 58.9 % (53/90) were confirmed to form biofilms on a polystyrene plate after 24 h, and the intensity of the biofilm production varied strongly between positive strains. Among the biofilm-producing isolates, 24.5 % (13/53) carried the icaA gene and 3.8 % (2/53) carried the bap gene. Among the isolates that did not produce biofilms, the icaA gene and bap gene were detected in 8.1 % (3/37) and 2.7 % (1/37) of isolates, respectively. This is the first report demonstrating that CNS isolated from feline conjunctiva can potentially be a bap gene reservoir. Preliminary comparison of the genetic profiles of three bap-positive isolates collected from cats showed that each of the isolates has a different genetic background with a high similarity with the human strain of S. epidermidis.     

Trace Metal Fluxes in a Sphagnum Peatland – Humic Lake System as a Consequence of Drainage

The study was conducted in a small (7.4 ha) peatland system with a humic lake concentrically surrounded by Sphagnum mat (Caricetum limosae) and Sphagnum bog (Ledo-Sphagnetum) in the Mazurian Lakeland in North-Eastern Poland. The peatland was situated in a forested catchment with a total area of 16.25 ha, and was influenced by surface drainage, artificially designed for forestry purposes. The content of zinc (Zn), cadmium (Cd), lead (Pb), copper (Cu) and hydrogen (H) ions was analysed in waters from precipitation, humic lake, mat, bog and surface outflow. The yearly budget of these ions, expressed as difference between atmospheric inflow and surface outflow, for the whole catchment was presented. The drainage of the peatland system over three years was accompanied by an increase in pH of lake, mat and bog waters. The whole catchment retention of H⁺ and trace metals studied decreased yearly in absolute values under these conditions. The peatland system began to leach Pb into surface waters and lost its ability to intercept Cd. These effects could be caused by intensity of outflow of water and sulphates and lowering of water storage. However, the potential to retain Zn and Cu ions was preserved.     

Some aspects of purinergic signaling in the ventricular system of porcine brain

Background

Numerous signaling pathways function in the brain ventricular system, including the most important - GABAergic, glutaminergic and dopaminergic signaling. Purinergic signalization system - comprising nucleotide receptors, nucleotidases, ATP and adenosine and their degradation products - are also present in the brain. However, the precise role of nucleotide signalling pathway in the ventricular system has been not elucidated so far. The aim of our research was the identification of all three elements of purinergic signaling pathway in the porcine brain ventricular system.

Results

Besides nucleotide receptors on the ependymocytes surface, we studied purines and pyrimidines in the CSF, including mechanisms of nucleotide signaling in the swine model (Sus scrofa domestica). The results indicate presence of G proteins coupled P2Y receptors on ependymocytes and also P2X receptors engaged in fast signal transmission. Additionally we found in CSF nucleotides and adenosine in the concentration sufficient to P receptors activation. These extracellular nucleotides are metabolised by adenylate kinase and nucleotidases from at least two families: NTPDases and NPPases. A low activity of these nucleotide metabolising enzymes maintains nucleotides concentration in ventricular system in micromolar range. ATP is degraded into adenosine and inosine.

Conclusions

Our results confirm the thesis about cross-talking between brain and ventricular system functioning in physiological as well as pathological conditions. The close interaction of brain and ventricular system may elicit changes in qualitative and quantitative composition of purines and pyrimidines in CSF. These changes can be dependent on the physiological state of brain, including pathological processes in CNS.     

Morphological and functional alterations in adult boar epididymis: Effects of prenatal and postnatal administration of flutamide

Background

The dynamic cross-talk between epididymal cells is hormonally regulated and, in part, through direct cell-to-cell interactions. To date, no information is available regarding possible impact of anti-androgens on the proteins involved in the gap junctional communication within the boar epididymis. Thus, a question arised whether prenatal or postnatal exposure to an anti-androgen flutamide alters the expression of gap junction protein - connexin43 (Cx43) and androgen receptor (AR) expression in the caput, corpus and cauda epididymis and leads to delayed effects on morphology and function of adult pig epididymis.

Methods

First two experimental groups received flutamide prenatally on gestational days 20-28 and 80-88 (GD20 and GD80) and further two groups were exposed to flutamide postanatally on days 2-10 and 90-98 after birth (PD2 and PD90). Epididymides were collected from adult boars. Routine histology was performed using hematoxylin-eosin staining. The expression of Cx43 and AR were analyzed using immunohistochemistry and Western blotting. Both analyses were supported by quantitative approaches to demonstrate the variations of the expression levels following the treatment. Apoptotic cells were identified using TUNEL assay.

Results

Histological examination revealed differences in epididymal morphology of flutamide-exposed boars when compared to controls. Scarce spermatic content were seen within the corpus and cauda lumina of GD20, PD2 and PD90 groups. Concomitantly, frequency of epididymal cell apoptosis was significantly higher (p < 0.05) after exposure to flutamide at GD20. Moreover, in GD20, PD2, and PD90 groups, significantly lower AR expression (p < 0.05) was found in the principal and basal cells of the corpus and cauda regions, while in the stromal cells AR expression was significantly reduced (p < 0.05) along the epididymal duct. Concomitantly, a decrease in Cx43 expression (p < 0.05) was noticed in the stromal cells of the cauda region of GD20 and PD2 groups. This indicates high sensitivity of the stromal cells to androgen withdrawal.

Conclusions

The region-specific alterations in the epididymis morphology and scarce spermatic content within the lumina of the corpus and cauda indicate that flutamide can induce delayed effects on the epididymal function of the adult boar by decrease in AR protein levels that results in altered androgen signaling. This may cause disturbances in androgen-dependent processes including Cx43 (de)regulation, however, we can not exclude the possibility that in response to flutamide decreased Cx43 expression may represent one mechanism responsible for functional disturbance of the boar epididymis.     

Kinetics of the response of four positive acute phase proteins in pigs experimentally infected with toxigenic Pasteurella multocida

In the present study, acute phase proteins (APP) responses in pigs after infection with toxigenic strain of Pasteurella multocida (Pm) were evaluated. Twelve piglets from a herd free from toxigenic Pm were used. Six of them were infected intranasally with Pm. CRP, Hp, SAA and Pig-MAP concentrations were measured using commercial ELISA tests. CRP and Hp were significantly induced from 2 days post inoculation (dpi). The concentration of Hp in inoculated pigs remained elevated until the end of the study. The concentrations of SAA and Pig-MAP increased significantly from 3 dpi, and remained elevated to 5 or 7 dpi, respectively. Strong correlations were observed between concentration of Hp or SAA and changes in the lungs. No correlations were found between levels of APP in serum and changes observed in the turbinates. On the basis of our investigation, we cannot state that evaluation of APP concentrations in serum may provide useful information about severity of atrophic rhinitis. However, the concentration of all investigated APP increased after inoculation. Thus, APP-measurements may reveal ongoing infection. Monitoring of APP concentrations in the pig herds may help to pinpoint infected animals even before clinical signs are present. If found, these "APP-high" animals may be chosen next for pathogen-specific diagnostics. Early diagnosis and therapy may prevent the infection from spreading in the herd. Moreover, monitoring of APP concentrations in serum may be useful for selecting clinically healthy pigs before integration into an uninfected herd. Future studies should focus on the possibility of distinguishing infected and non-infected pigs under field conditions.     

Regional Pattern of Heavy Metal Content in Lake Sediments in Northeastern Poland

We investigated sediments from 23 lakes situated in northeastern Poland and analyzed them for major constituents and selected heavy metals. Short sediment cores were collected from the deepest parts of the lakes, and subsequently, a surface layer (0-2?cm) and reference layer (50-52?cm) were sampled from each. In the collected samples, the content of the major constituents (organic matter, carbonates, and minerogenic material) and chosen heavy metals (Cd, Cu, Ni, Pb, and Zn) was analyzed. In the reference layer, representing natural metal content, we identified quite a substantial diversity among lakes, making it difficult to pinpoint one geochemical background value for the whole region. A multivariate analysis of the interrelationships among elements and a comparison of the median values revealed no statistically significant differences between surface and reference levels. The ratio of the mean content in the surface and reference sediments ranged from 0.9 to 1.6, indicating the lack of or only slight anthropogenic pollution in surface sediments. From a spatial perspective, higher metal contents were observed in the eastern part of the study area, but this trend manifested in both surface and reference sediments. Thus, the inference is that the recently accumulated sediments are characterized by a content that is representative of the natural geochemical background for the selected metals.