The paper reviews the various factors affecting dystocia in cattle. It is based mainly on the recent studies found in the literature of the subject but refers occasionally to some older papers as well. The factors are grouped into four main categories: direct factors, phenotypic factors related to calf and cow, non-genetic and genetic factors. The first group includes malpresentations and uterine torsion. The second one includes: calf birth weight, multiple calvings, perinatal mortality, cow pelvic area, cow body weight and body condition at calving, gestation length. The non-genetic factors are: cow age and parity, year and season of calving, place of calving, maintenance practises, disorders, calf sex and nutrition. Other non-genetic factors are the level of hormones in the periparturient period, in vitro production of embryos and embryo cloning. Finally, the genotypes of cow, bull and calf, inbreeding, muscular hypertrophy, selection and quantitative trait loci form the fourth group of genetic factors.

We examined molecular events and morphological features associated with apoptosis induced by anthraquinone anticancer drugs aclarubicin, mitoxantrone and doxorubicin in two spontaneously immortalized cell lines (NIH 3T3 and B14) in relation to cytotoxicity of these drugs. The investigated cells showed similar sensitivity to aclarubicin but different sensitivity to doxorubicin and mitoxantrone: mitoxantrone was the most cytotoxic drug in both cell lines. All three drugs triggered both apoptosis and necrosis but none of these processes was positively correlated with their cytotoxicity. Apoptosis was the prevalent form of cell kill by aclarubicin, while doxorubicin and mitoxantrone induced mainly the necrotic mode of cell death. The extent and the timing of apoptosis were strongly dependent on the cell line, the type of the drug and its dose, and were mediated by caspase-3 activation. A significant increase in caspase-3 activity and the percentage of apoptotic cells, oligonucleosomal DNA fragmentation, chromatin condensation and formation of apoptotic bodies was observed predominantly in B14 cells. NIH 3T3 cells showed lesser changes and a lack of DNA fragmentation. Aclarubicin was the fastest acting drug, inducing DNA fragmentation 12 h earlier than doxorubicin, and 24 h earlier than mitoxantrone. Caspase-3 inhibitor Ac-DEVD-CHO did not show any significant effect on drug cytotoxicity and DNA nucleosomal fragmentation.

In the present study, the analysis of epithelial cells derived from various sources was undertaken, beginning from the mammary gland tissue through the primary cultures and their subsequent passages. The objective of the study was the comparative analysis of the stage in which the epithelial cells obtained from individuals in different lactation cycles and disparate phases of cell culture growth are the most suitable for morphological research and analysis of gene expression activity. The cultures of primary bovine mammary epithelial cells and passages were identified morphologically using immunocytochemical methods. After positive identification, real-time PCRs were performed for the analysis of the expression level of casein genes, whey protein genes, and butyrophilin gene. The most stable reference genes in real-time PCRs for the mammary gland tissue and cell cultures were also determined. Of the reference genes, the UXT and GAPDH genes appeared to be the most stable ones for the mammary gland tissue samples and epithelial cell cultures. The results obtained allowed concluding that the mammary gland samples collected from heifers constituted the most effective material for the initiation of primary cultures. The primary cultures formed characteristic for the mammary gland tissue dome structures, which images were obtained using confocal microscopy. The highest levels of expression of the CSN1S1, CSN1S2, CSN2, and CSN3 genes were detected in primary cultures. The levels of expression of whey protein genes (LALBA and BGL) were highest in the second passage. The most abundant expression of the BTN1A1 gene was observed in primary cultures and the third passage. On the basis of the whole experiment, it can be concluded that primary cultures and cells of the second passage derived from heifer individuals appeared to be the best materials for the analysis of mammary gland function and gene expression activity.

The novel approach for deposition of iron oxide nanoparticles with narrow size distribution supported on different sized graphene oxide was reported. Two different samples with different size distributions of graphene oxide (0.5 to 7 μm and 1 to 3 μm) were selectively prepared, and the influence of the flake size distribution on the mitochondrial activity of L929 with WST1 assay in vitro study was also evaluated. Little reduction of mitochondrial activity of the GO-Fe3O4 samples with broader size distribution (0.5 to 7 μm) was observed. The pristine GO samples (0.5 to 7 μm) in the highest concentrations reduced the mitochondrial activity significantly. For GO-Fe3O4 samples with narrower size distribution, the best biocompatibility was noticed at concentration 12.5 μg/mL. The highest reduction of cell viability was noted at a dose 100 μg/mL for GO (1 to 3 μm). It is worth noting that the chemical functionalization of GO and Fe3O4 is a way to enhance the biocompatibility and makes the system independent of the size distribution of graphene oxide.

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Szatkowska, I., Dybus, A., Grzesiak, W., JFdrzejczak, M. and Muszyliska, M. 2009

The aim of this study was to examine the effect of three structurally different anticancer drugs-the pro-oxidative anthracyclines doxorubicin (DOX) and aclarubicin (ACL), and antioxidative anthraquinone mitoxantrone (MTX) on the fluidity of plasma membrane of immortalized rodent fibroblasts using fluorescence spectroscopy and electron spin resonance (ESR) techniques. Two kinds of fluorescent probes (TMA-DPH and 12-AS) and spin labels (5-DS and methyl-12-DS) were used to monitor fluidity in the hydrophobic core and in the polar headgroup region of the lipid bilayer. Immortalized hamster B14 and NIH 3T3 mouse fibroblasts were exposed to DOX, ACL and MTX. We demonstrate that these drugs influence predominantly the hydrophobic core of the lipid bilayer, inducing significant decrease in its fluidity at low concentrations (2-5 microM). A decreased membrane fluidity at the surface of the lipid bilayer was observed only at a higher concentration (20 microM) of the drugs, which indicates that DOX, ACL and MTX intercalate mainly into the hydrophobic core of the membrane, thereby perturbing its structure.

The aim of this study was to relate polymorphic variants of CYP19, CYP21 and ER1 genes to reproductive traits in 472 Polish Holstein-Friesian cows. High frequencies of one of the homozygous genotypes were found. The ER1/SnaBI AA homozygotes were not identified. In the first and third lactation, an average calving-to-conception interval (CLVC) in cows of ER1/SnaBI GG genotype was significantly shorter (P ≤ 0.05) than in heterozygous cows. In the cows of ER1/ BglI GG genotype, significantly shorter CLVC (P ≤ 0.05) was observed compared to heterozygotes in the first lactation, whereas in the third lactation, CLVC in homozygous cows was significantly longer (P ≤ 0.05) than in heterozygous ones. It was also found that homozygous cows were characterized by significantly longer calving interval (CLVI; P ≤ 0.05) compared to heterozygotes in the third lactation. Longer CLVCs in CYP19 AA cows were found, compared to heterozygotes, and this difference was significant in the first and third lactation (P ≤ 0.05). Similarly, the average CLVIs were longer in CYP19 AA homozygotes than in heterozygous cows; however, significance was proven only in the third lactation (P ≤ 0.05). Description of the molecular mechanisms regulating reproduction, and thus identification of the individuals of genotypes with optimal potential may facilitate the employment of selected reproductive model by a breeder. Gene polymorphism, calving interval, calving-to-conception interval, pregnancy length, inseminationsAbbreviations: CYP19 -cytochrome P450 aromatase gene, CYP21 -steroid 21-hydroxylase gene, ER -oestrogen receptor gene, CLVC -calving-to-conception interval, CLVI -calving interval, INSEM -number of services per successful conception, PREG -pregnancy length.Genetic improvement of the reproductive traits in dairy cattle is crucial not only for ensuring quantitative aspect of production but mainly for the efficient course of lactogenesis and lactopoesis. It is an extremely difficult process, regulated at many stages in a multifactorial manner. Moreover, coefficients characterizing reproductive traits are of low heritability and there is a small number of described genetic markers associated with reproduction that could be used as potential factors supporting the classical selection methods. Therefore, searching for such markers is admittedly important both for the application and research purposes. So, the analysis of the influence of different polymorphic variants of enzymes crucial for oestrogen synthesis on the reproductive traits or of protein factors mediating their actions seems fully justified.The reproductive cycle of all mammalian females is regulated mainly by oestrogen hormones. They are synthesized through the aromatization of androgens, which, in females, are mainly of ovarian origin. In this complex process, a key role is played by the p450 aromatase, which is an enzymatic complex comprising two proteinsnonspecific microsomal flavoprotein reductase and specific haemoglycoprotein, that is, cytochrome P450 aromatase, coded f...