Analysis of integrins and vascular endothelial growth factor isoforms mRNA expression in the canine uterus during perimplantation period

Integrins are the major receptors within the extracellular matrix (ECM) that mediate several functions connected with cell life and metabolism, such as cell adhesion, migration, cytoskeletal organization, proliferation, survival, and differentiation. A vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. It has been suggested that the expression of this gene may play crucial physiological roles in reproductive organs. All investigated endometrial tissues were isolated on day 10-12 after mating. Control bitches, used in this study, were in metestrus, which was determined according to the vaginal cytology and progesterone level in blood. Early pregnancy was verified by flushing the uterine horns with PBS. Total RNA was isolated from the bitches endometrium by means of the Chomczyński and Sacchi method, treated by DNase I, and reverse-transcribed into cDNA. A quantitative analysis of integrins alpha2b, beta2 and beta3, VEGF 164, 182 and 188 cDNA was performed by RT-PCR. In results we have shown an increased expression of all investigated genes (integrins alpha2b, beta2 and beta3, VEGF 164, 182, and 188) in pregnant bitches uterus as compared to non-pregnant females (P < 0.001). Our results indicated that the expression of genes encoding integrins and vascular endothelial growth factors is different in relation to the time of the embryo implantation and it is increased in the first period of this process. This may be associated with the induction of specific mechanisms responsible for receptivity of uterus following the embryo attachment. In addition, all of investigated genes are up-regulated in a pregnancy-specific manner and the increased expression of these genes may regulate the uterus function during the implantation of canine embryos.     

Phylogenetic similarity of the canine parvovirus wild-type isolates on the basis of VP1/VP2 gene fragment sequence analysis

Biological material was taken from dogs with diarrhoea. Faecal samples were taken from within live animals and intestinal tract fragments (i.e. small intestine, and stomach) were taken from dead animals. In total, 18 specimens were investigated from dogs housed alone or in large groups. To test for the presence of the virus, latex (On Site Biotech, Uppsala, Sweden) and direct immunofluorescence tests were performed. At the same time, polymerase chain reaction (PCR) with primers complementary to a conservative region of VP1/VP2 was carried out. The products of amplification were analysed on 2% agarose gel. The purified products were cloned with the Template Generation System (Finnzymes, Espoo, Finland) using a transposition reaction and positive clones were searched using the 'colony screening by PCR' method. The sequencing gave 12 sequences of VP1/VP2 gene fragments that were of high similarity. Among the 12 analysed sequences, six exhibited 88% similarity, four exhibited 100% similarity and two exhibited 71% similarity.     

Genotype-by-environment interaction of barley DH lines infected with Fusarium culmorum (W.G.Sm.) Sacc.

Infection of plants by pathogens is a biotic environmental stress. Barley plants are infected, among others, by Fusarium culmorum—a pathogen affecting seedling, head, root and stem. The infection can result in reduced yield and grain quality. The aim of the study was to compare the reaction of inoculated and non-inoculated barley doubled haploids (DHs) with F. culmorum in various environments. Thirty-four genotypes were inoculated with an isolate of F. culmorum. The experiment was carried out over 6 years. Kernel weight per spike, 1000-kernel weight and percentage of plump kernels were observed in control and inoculated plants. Genotype-by-environment (GE) interaction and its structure with reference to the environments and genotypes were analysed. Additional information about the sensitivity of healthy and infected genotypes to environments was determined by the regression analysis. Statistical computation was made using the SERGEN software. Lines were considered as unstable when their GE interaction was significant at P = 0.05. Unstable genotypes were classified as intensive or extensive according to the results of the regression analysis. It was found that infection with Fusarium decreased the stability of barley lines in different environments. Interaction of unstable infected genotypes with environments, most often, could not be explained by the regression—their response to various environmental conditions appeared to be unpredictable. Selection of lines less susceptible to biotic and abiotic stresses was possible due to comparison of classification of healthy and infected lines, which was made based on their main effects and GE interaction.     

Rye Flour Extraction Rate Affects Maillard Reaction Development,Antioxidant Activity,and Acrylamide Formation in Bread Crisps

This report shows the effect of rye flour extraction rate on Maillard reaction, antioxidant activity, and acrylamide formation during toasting of rye bread crisps. Four rye flours with extraction rates of 70, 85, 95, and 100% were tested. Maillard reaction development was studied by measuring browning development, hydroxymethylfurfural (HMF), and glucosilisomaltol (GIM) formation, as well as antioxidant activity. Results showed that HMF and GIM concentrations in toasted bread crisps were higher as the flour extraction rate increases. Antioxidant activity increased during toasting as a consequence of antioxidant Maillard reaction product formation. Acrylamide concentration was clearly affected by free asparagine content of flour, while no effect of dietary fiber and natural antioxidant content of flours had an effect on acrylamide formation. Overall data suggest that the rate of Maillard reaction is higher in whole flours because of their higher free amino acid and protein content.     

The effect of the source and dosage of dietary Cu on redox status in rat tissues

The aim of this experiment was to investigate whether the amount of Cu added to the diet of rats can be reduced without adversely affecting the antioxidant status of tissues and growth, and whether copper nanoparticles can be used for this purpose. For four weeks, four experimental groups of rats were fed diets with two dosages of added Cu (standard—6.5 or 3.25 mg/kg) in two forms (standard—CuCO₃ or copper nanoparticles). Replacing the CuCO₃ supplement with CuNPs resulted in a decreased lung weight and an increased Cu content in brain, kidney and lung, intensification of lipid peroxidation processes, and weakened antioxidant defence in the lungs and kidneys. This treatment also reduced the Cu content in heart, level of lipid oxidation in the liver and testes and improved antioxidant defence in the brain. Reducing the addition of Cu to the diet from 6.5 to 3.25 mg/kg reduced lung weight and increased lipid peroxidation in the liver, heart and lungs, and also weakened antioxidant defence in the lungs and testes. This treatment also weakened the lipid peroxidation process in the spleen, small intestine and brain and strengthened the antioxidant defence of the brain and kidneys. In conclusion, replacing CuCO₃ with CuNPs and reducing the level of Cu in the diet of rats has a particularly unfavourable effect on the respiratory system, causing adverse changes in the lungs. However, these treatments have a clearly positive effect on the redox status of the liver and brain.     

Inhibition of Armillaria and Heterobasidion growth by Penicillium adametzii isolated from Pinus sylvestris forest soil

Penicillium adametzii significantly inhibited the growth of Heterobasidium annosum sensu stricto on 2% potato dextrose agar after 10 days at 20–25°C. Chloroform extracts from P. adametzii culture filtrate and from culture grown on rice often decreased the dry weight of Armillaria colony and diameter of Heterobasidion colony in vitro. The effect depended on species and isolate of Armillaria, Heterobasidion and P. adametzii and concentration of the extract. Higher concentration of chloroform extract increased the dry weight of stems and roots of Pinus sylvestris plants inoculated with either of two isolates of Armillaria ostoyae and one isolate of Armillaria gallica. Lower concentrations of chloroform extract increased the dry weight of stems and roots of P. sylvestris inoculated with another isolate of A. gallica. Culture filtrate of P. adametzii decreased the length of necrosis on P. sylvestris stem inoculated with either of two of four isolates of H. annosum s.s., but if applied 1 month before inoculation, it increased the length of necrosis caused by another H. annosum s.s. isolate. Twenty fractions from the P. adametzii chloroform extract were separated by thin layer chromatography. Carboxylic acids, flavonoids and glycosides were detected in chloroform extract of P. adametzii by high performance liquid chromatography/mass spectrometer analysis. This is the first report of interactions between P. adametzii and either Armillaria or Heterobasidion with P. sylvestris.     

Zinc tolerance in the ant species Myrmica rubra originating from a metal pollution gradient

This study was designed to test whether the ant species Myrmica rubra had developed a tolerance to metal pollution. Larvae and workers, collected from seven sites located along a metal pollution gradient in the vicinity of Olkusz, Poland were exposed to a range of Zn concentrations in food, artificially contaminated in a laboratory experiment. The mortality of adults and larvae, as well as maximum larvae body mass, was recorded. The results of this study indicate that larvae mortality was independent of the site pollution, but decreased significantly with the Zn concentration in food. Opposite results were found for workers: irrespectively of Zn concentration in the food, the mortality decreased with site pollution, indicating enhanced metal tolerance in workers. Contrary to the expectations, the maximum larvae mass estimated with the von Bertalanffy curve, increased by the pollution gradient. Larvae were more sensitive to Zn than the adults, but their higher mass at the most polluted sites, may have enhanced tolerance to metal pollution.     

Immunobiology of bovine mammary gland: apoptosis of somatic cells in milk during naturally occurring mastitis

Mastitis remains a major cause of economic losses in dairy herds. It is believed, that the enhancement of natural defense mechanisms in mammary gland may be helpful in the treatment of bovine mastitis. Our study was designed to assess the apoptosis of leukocytes isolated from bovine milk during subclinical staphylococcal mastitis. Milk samples were collected from cows naturally infected with one pathogen--Staphylococcus aureus and from animals with mastitis caused by several pathogens, including S. aureus. It has been determined that the rate of apoptosis was lower in mastitic milk, as compared with control samples, although varied significantly between groups. High percentage of apoptotic cells was detected in milk with high counts of pathogenic bacteria. In all groups the rate of apoptosis was dependent on the bacterial load, although various correlations were identified. Thus, it is postulated, that the rate of apoptosis of somatic cells in mastitic milk is related to the etiology of infection and is determined by the bacterial load.     

Epidemiological features of Morel's disease in goats

Morel's disease caused by Staphylococcus aureus subsp. anaerobius was diagnosed for the first time in Poland in October 2006 in a goat flock. A second infected flock was found two months later. The course of the disease in both flocks was observed for 15-17 months. Clinical manifestation was confined to abscesses located near major superficial lymph nodes, mostly: superficial cervical, subiliac, parotid and mandibular. At necropsy no other lesions were found. The incubation period was estimated at 3 weeks. Clinical signs were seen both in young and adult goats and up to 7 abscesses in one animal were noted. Abscesses tended to persist for 1 to 5 months, then rupture and heal completely. The initial high in-flock point prevalence in both flocks (93.6% and 84.4%) dropped to approximately 10-30% during next 3-4 months. Until the end of the observation period the in-flock point prevalence remained at this level and only single abscesses were observed, mainly in young animals. No influence of the concurrent caprine arthritis encephalitis virus (CAEV) infection on the clinical course of Morel's disease was noticed. It is to be concluded that the clinical course of Morel's disease in a goat flock resembles caseous lymphadenitis (CLA). However, in Morel's disease abscesses occur more frequently in young goats and are located near, not inside, the lymph nodes, as in the case with CLA. Also, the incubation period of Morel's disease seems to be shorter (3 weeks versus 2-6 months in CLA).     

Occurrence of <Emphasis Type="Italic">Puccinia</Emphasis> spp. spores in Madeira Island and their phytopathological importance

The genus Puccinia represents rust infections, which are responsible for great productivity losses in crops of commercial and ornamental plants worldwide. This work is aimed at determining the occurrence of Puccinia spp. spores in Madeira Island in order to infer the exposure risks from a phytopathological point of view. A phytopathological analysis was performed in 203 local plant samples between January 2003 and December 2012. During the same period the airborne concentrations of rust spores were monitored following well-established guidelines. Aerobiological data was compared with meteorological records. Based on macro and microscopic analyses, five species of rusts were identified: P. horiana, P. buxi, P. porri, P. pelargonii-zonalis, and P. sorghi, and they were found mostly in spring and summer. A total of 20 samples out of 203 analysed plants (9.8%), were infected with Puccinia spores, i.e., P. horiana (5.9%), P. buxi (1.47%), P. porri and P. pelargonii-zonalis (0.98%), and P. sorghi (0.5%). During the studied period Puccinia spores attained an annual average concentration of 126 spores m^?3 and most of them were recorded between March and October. Meteorological factors were determinant in fluctuations in spore concentration. Relative humidity was the parameter that favoured the biggest release and dispersal of the rust spores, whereas rainfall revealed a significant negative effect. Rusts do not represent an important plant pathogen in Madeira Island, as shown by the low infection frequencies and levels of airborne spore concentrations.