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Rozwój infrastruktury wiejskiej oznacza dla mieszkańców wsi nowe możliwości zaspokajania potrzeb. Wykorzystanie tych możliwości spowodowało zmianę poziomu i struktury ich wydatków. W szerszym stopniu korzystano z udogodnień infrastrukturalnych i prywatnych środków transportu. Większą uwagę zwracano na wygodę życia. Coraz więcej mieszkań było modernizowanych i wyposażanych w instalacje wodociągowe i grzewcze. Zwiększyło się uczestnictwo w kulturze, wypoczynku i rekreacji.
Исследовали сыворотку и кишечный слизь поросят категории SPF, инфицируемых внутрикишечно, внутрибрюшнно либо в пупочную вену различными штаммами ротавирусов поросят и телят (OSU, RFC и RCI). Показали возбуждение местного синтеза интерферона в кишке. Не отметили зависимости между образованием интерферона в тонкой кишке и уровнем интерферона в сыворотке. Присутствие интерферона в слизи тонкой кишки показали через 24—55 ч.ч. после введения ротавируса в кишку или пупочную вену.
110 blood samples from clinically healthy mares of English breed, half-blood and the Wielkopolska race were tested for the presence of EHV-1 and EHV-2 using nested PCR. 15 samples were EHV-1-positive, 24 samples were EHV-2-positive, whereas only 4 samples were both EHV-1 and EHV-2 positive. The virus was isolated from PBLs in equine dermal cell cultures by co-cultivation or by culture inoculation with cell lysates derived from PBLs. A total of 14 strains were isolated from EHV-2 and EHV-1/EHV-2-positive samples. However, all of them were identified by nPCR as being type 2. Since no EHV-1 was isolated, even from dually infected leukocytes, it was concluded that the presence of EHV-2 does not stimulate in-vitro isolation of EHV-1 from infected leukocytes. It is tempting to speculate that such stimulatory effects in-vivo may involve EHV-2-induced immunosupression. Despite any possible mechanism of EHV-1 stimulation, it seems that EHV-2 does not play a significant role in the epidemiology of EHV-1-caused miscarriages in horses since mixed infections are rather rare.
The aim of the study was to investigate the influence of equine herpesvirus type 1 (EHV-1) infection on actin cytoskeleton in ED (equine dermal). Cells in vitro.ED cells were infected with a strain of Jan-E of EHV-1, fixed and stained for the presence of actin and virus antigen. Results were evaluated by confocal microscopy. The assembly of the actin cytoskeleton was heavily disturbed. In order to affirm whether changes in cytoskeleton of EHV-1 infected cells depend on the type of cells, we infected Vero cell culture with 2 different standard strains of EHV-1 - Rac-H, AIV - and Jan-E isolated from an aborted fetus. Unfortunately, the infection of Vero cells with the strain Jan-E of EHV-1 failed because this strain was not adapted to the heterogeneous cell line. Only strain Rac-H and AIV can replicate in Vero cells, which was determined through the application of PCR.
The nested PCR technique (Borchers and Slater, 1993) was applied for the diagnosis of EHV-1 infections. DNA samples were isolated from livers of miscarried fetuses or dead foals. After amplification we detected EHV-1 specific sequences in 9 out of 15 fetuses and in 2 out of 6 foals. PCR results were compared to results of routinely performed immunofluorescent detection of viral antigen in cryosections of tissues. Six tested fetuses were FAT-positive, 7 FAT-negative and 2 were questionable. One FAT-negative fetus and two questionable were PCR-positive. All foals were FAT-negative. We have demonstrated that PCR is more sensitive than FAT and allows verification of inconclusive results of antigen detection. PCR is also suitable for detection of EHV-1 specific sequences in archival samples frozen up to 3 years.
In previous experiments, we have demonstrated that the presence of equine herpesvirus 2 (EHV-2) enhanced plaque formation in cell cultures infected with equine herpesvirus type 1. To determine whether a specific region of the EHV-2 genome is responsible for this effect, we have constructed a library of Bam HI fragments of the EHV-2 genome ligated into pcDNA plasmid. Equine dermal (ED) cell cultures were subsequently transfected with the constructs, passaged 5 times, tested for the presence of the plasmids and infected with EHV-1 at MOI=0.01. Only in cultures transfected with the pcDNA/Bam HI[G]construct, designated Δ2/4, the mean number of plaques at 24 hrs p.i. was approximately 10 times higher than in non-transfected controls. Virus titers in culture supernatants as well as in freeze-thawed cells were 4- and 5-fold higher, respectively, than in non-transfected cultures. These differences were observed only at 24 hrs p.i. At 48 hrs p.i. cultures were completely destroyed and, surprisingly, the virus titer was slightly lower in the supernatant of transfected cells. However, the titer of EHV-1 in freeze-thawed culture was exactly the same as in the control. These results suggest that the presence of Bam HI[G] fragment of the EHV-2 genome stimulates (accelerates?) plaque formation only at earlier stages of infection but does not influence the total yield of EHV-1 at 48 hrs p.i. The exact mechanism of this stimulation remains unclear and further experiments are necessary to determine the role of putative EHV-2 proteins encoded by Bam HI [G] fragment of the EHV-genome.
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