Phyto- and endogenous estrogens differently activate intracellular calcium ion mobilization in bovine endometrial cells

The main purpose of this study was to check whether phyto- and endogenous estrogens influence calcium ion mobilization [Ca(2+)](i) in bovine endometrial cells and whether this action is connected with biological effects i.e. prostaglandin (PG)F(2alpha) production. In our study we used two calcium measurement methods by comparing the microscopic method with widely used quantitative - spectrofluorometric method of [Ca(2+)](i) measurement. We also wanted to confirm whether visualization of calcium ion [Ca(2+)](i) in cells using microscopic method supported by micro image analysis (Micro Image Olympus system) reflects real, qualitative changes in the ion concentration. In both methods a cell-permeable form of fluorescent [Ca(2+)](i) indicator Fura-2 was used. Cultured bovine endometrial epithelial and stromal cells influenced by phorbol-2-myristate-13-acetate (PMA; positive control), estradiol 17-beta (E(2); endogenous estrogen) and active metabolites of phytoestrogens (environmental estrogens) were used as a model to study PGF(2alpha) secretion and [Ca(2+)](i) mobilization in the cells. Equol and para-ethyl-phenol in doses of 10(-8)-10(-6) M increased PGF(2alpha) concentration both in epithelial and stromal cells (P<0.05). In both methods, equol and para-ethyl-phenol did not cause intracellular [Ca(2+)](i) mobilization in epithelial and stromal cells (P>0.05). Both methods revealed that only E(2) and PMA induced intracellular [Ca(2+)](i) mobilization in epithelial and stromal cells (P<0.05). The results of both methods were highly correlated (P<0.001; r=0.82 for epithelial cells and r=0.89 for stromal cells). In conclusion, both methods gave approximately the same results and showed that phytoestrogens, in contrast to PMA and E(2), did not cause intracellular [Ca(2+)](i) mobilization in endometrial cells. The obtained results proved that the [Ca(2+)](i) visualization method supported by micro image analysis can produce similar results to the spectrofluorometric method.     

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.     

Requirement for hippocampal CA3 NMDA receptors in associative memory recall

Pattern completion, the ability to retrieve complete memories on the basis of incomplete sets of cues, is a crucial function of biological memory systems. The extensive recurrent connectivity of the CA3 area of hippocampus has led to suggestions that it might provide this function. We have tested this hypothesis by generating and analyzing a genetically engineered mouse strain in which the N-methyl-D-asparate (NMDA) receptor gene is ablated specifically in the CA3 pyramidal cells of adult mice. The mutant mice normally acquired and retrieved spatial reference memory in the Morris water maze, but they were impaired in retrieving this memory when presented with a fraction of the original cues. Similarly, hippocampal CA1 pyramidal cells in mutant mice displayed normal place-related activity in a full-cue environment but showed a reduction in activity upon partial cue removal. These results provide direct evidence for CA3 NMDA receptor involvement in associative memory recall.     

Biological activity of alpha-galactoside preparations from Lupinus angustifolius L. and Pisum sativum L. seeds.

Biological activity tests were performed on alpha-galactoside preparations obtained from Lupinus angustifolius L. cv. Mirela (alkaloid-rich) and Pisum sativum L. cv. Opal seeds. The studies included the following tests: acute toxicity, cytotoxic test, delayed type hypersensitivity (DTH), plaque-forming cell number (IgM-PFC), and influence on the growth of bifidobacteria and coliform presence in rat colon. Results of these studies showed that alpha-galactosides from lupin and pea seeds were essentially nontoxic. Their acute toxicity (LD(50)) in mice was >4000 mg kg(-1) of body weight. alpha-galactoside preparations were not cytotoxic for mouse thymocytes in vitro. The in vitro test shows that oligosaccharides from lupin and pea are utilized by selected beneficial colon bacterium strains. The in vivo experiment demonstrated that alpha-galactosides from legume significantly influenced the growth of bifidobacteria in rats colon. Simultaneously, the decrease of the coliform presence was observed. The chemical composition of the tested preparations had no significant effect on their biological activity.     

Lead Partitioning in the Bottom Sediment of Rybnik Reservoir (Southern Poland)

Total concentration of lead and its individual partitioning, i.e. exchangeable, adsorbed, organically bound, carbonate, sulfide and residual have been assayed in the bottom sediment of Rybnik reservoir applying AAS. The contribution of particular fractions to the total lead concentration was as follows: carbonates (37%) > sulfides (28%) > residual (14%) > adsorbed (10%) > exchangeable (8%) > organic bonds (3%). The multiple regression analysis applied led to the conclusion that the increase in lead content in the bottom sediment resulted from an increase in poorly mobile forms, notably carbonates.     

The effect of feeding the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on cell-mediated immunity and protection against Yersinia ruckeri in pikeperch (Sander lucioperca)

The present study examined the influence of leucine metabolite β‐hydroxy‐β‐methylbutyrate (HMB), a natural product HMB, on nonspecific cell‐mediated defence mechanisms and protection against enteric redmouth disease (ERM) in pikeperch (Sander lucioperca). β‐hydroxy‐β‐methylbutyrate was fed in a pelletted ration at 50 mg kg^?1 of the feed for 8 weeks. The phagocytic ability and potential killing activity of blood and pronephric phagocytes were examined in HMB‐ and control‐fed fish before and after 8 weeks of feeding HMB. Simultaneously, the proliferative response of blood and pronephric lymphocytes stimulated by mitogens concanavaline A and lipopolisaccharide were examined in experimental and control groups. Following 8 weeks of HMB feeding, a challenge test was performed by injecting the fish with live pathogenic bacteria Yersinia ruckeri. β‐hydroxy‐β‐methylbutyrate applied in the diet for 8 weeks prompted a statistically significant (P<0.05) increase in phagocytic ability and potential killing activity of the blood and pronephric phagocytes and the proliferative response of blood and pronephric lymphocytes. The changes in these mechanisms correlated with protection against Y. ruckeri, the ERM pathogen. The results showed that feeding HMB increased the nonspecific cell‐mediated defence mechanisms and protection against ERM by reducing cumulative mortality (30%) following the challenge with pathogenic bacteria. Future studies will include determination of optimal doses and protocols of oral application of HMB to maximize the immunomodulatory effects and protection against viral diseases in intensive pikeperch culture.     

Skin mucus of Cyprinus carpio inhibits cyprinid herpesvirus 3 binding to epidermal cells

ABSTRACT: Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a mortal and highly contagious disease in common and koi carp. The skin is the major portal of entry of CyHV-3 in carp after immersion in water containing the virus. In the present study, we used in vivo bioluminescence imaging to investigate the effect of skin mucus removal and skin epidermis lesion on CyHV-3 entry. Physical treatments inducing removal of the mucus up to complete erosion of the epidermis were applied on a defined area of carp skin just before inoculation by immersion in infectious water. CyHV-3 entry in carp was drastically enhanced on the area of the skin where the mucus was removed with or without associated epidermal lesion. To investigate whether skin mucus inhibits CyHV-3 binding to epidermal cells, tail fins with an intact mucus layer or without mucus were inoculated ex vivo. While electron microscopy examination revealed numerous viral particles bound on the fins inoculated after mucus removal, no particle could be detected after infection of mucus-covered fins. Finally, anti-CyHV-3 neutralising activity of mucus extract was tested in vitro. Incubation of CyHV-3 with mucus extract reduced its infectivity in a dose dependent manner. The present study demonstrates that skin mucus removal and epidermal lesions enhance CyHV-3 entry in carp. It highlights the role of fish skin mucus as an innate immune protection against viral epidermal entry.     

Effect of teleost pituitary gonadotropins on the in vitro maturation of carp oocytes

Studies were conducted on the effects of various kinds of teleost pituitary gonadotropins on the in vitro maturation of carp oocytes. It was ascertained that after 24 h of incubation with gonadotropins, the highest percentage of oocytes with the nucleus in the migrating or peripheral position, or of mature oocytes, was observed in the case of ciprinide gonadotropins.Centrifuged hypophysial homogenates of the pike, sea-trout and pike-perch displayed a considerably weaker activity. The results obtained are indicative of a specificity of teleost pituitary gonadotropins.     

Length of winter coat in horses depending on husbandry conditions

This paper analyzes changes in the length of coat on selected body areas in horses and ponies kept under different husbandry (stable) conditions during the winter‐spring period. The study included 12 Ma?polski geldings and 12 geldings of Felin ponies aged 10–15 years. Horses were kept in two stables (six horses and six ponies in each stable). The type of performance, husbandry conditions and feeding of the studied animals were comparable. As of December 1, samples of hair coat from the scapula, sternum, back and abdomen areas of both body sides were collected seven times. The lengths of 20 randomly selected hair fibers were measured. Daily measurements of air temperature in the stables were also taken. An analysis of variance (ANOVA) was performed using the following factors: the body part from where the coat was sampled, the subsequent examination and the stable as well as the interaction between these factors. The significance of differences between means was determined with a t‐Tukey test. The relations between air temperature in the stable and hair length were calculated using Pearson's correlation. It was found that air temperature in the stable impacts the length of winter coat in horses and ponies. The effect of this factor is more pronounced in ponies; as in the stables with lower temperatures it produces a longer hair coat which is more evenly distributed over the body in comparison with horses. Keeping horses and ponies in stables with a low air temperature accelerates coat shedding by approximately 25 days. Coat shedding begins from the scapula area.