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β-N-acetylhexosaminidase was prepared from a liver lysosomal fraction obtained from rats between 18 days of gestation (group I) and 72 weeks of age (groups II-VI). A glycan chain analysis was performed after an electrophoresis and blotting, followed by a very sensitive detection system with highly specific digoxigenin-labelled lectins. The presence of high-mannose /hybrid type glycans, as well as their fucosylated forms was shown in all the experimental groups. Complex-type glycans with terminal sialic acid or galactose were present in all the groups except for 1-week-old rats in which only a positive reaction with lectins from Galanthus nivalis and Aleuria aurantia - was observed. Thus it may be assumed that age-related changes in the glycosylation pattern occur on the first days after birth.
A glycan chain analysis of the total oligosaccharide pool derived from rat liver arylsulfatase B was carried out by. P4 Gel Permeation Chromatography and sequential exoglycosidase digestion. It was found that 71% of rat liver arylsulfatase B oligosaccharides were sialylated. The relative contribution of particular structures in the total glycan pool was as follows: sialylated biantennary complex type glycans with terminal galactose--65%, high-mannose type glycans--15%, biantennary complex type glycans with core fucose and terminal N-acetylglucosamine--5%, O-linked oligosaccharides--3.5%.
The aim of the study was to determine the effect of pelleted cereal-based feed used in the diet for antelope sitatunga ( ) on feed intake, eating behaviour, rumination and nutrient digestibility. Three male sitatunga were fed a basal diet (chopped dehydrated lucerne, ground cereals, soyabean meal, vegetables and fruits) with access to meadow hay. The animals were allocated to 1 of 3 treatments according to 3 × 3 Latin square design and fed diets where 0, 50 or 100% of dry matter (DM) from basal diet was replaced with a pelleted cereal-based feed. DM intake of the basal diet and meadow hay, as well as overall DM intake and organic matter (OM) intake did not differ between treatments ( > 0.05). The eating rate (g DM · min–1) of the basal diet increased ( = 0.03) whereas eating time (min · day–1) and eating frequency (n · day–1) tended to decrease ( ≤ 0.07) linearly as pelleted feed inclusion in the diet increased. On the other hand, a tendency ( = 0.07) to longer time of hay intake (min · day–1) was observed with increasing inclusion of pelleted cereal-based feed in the basal diet. Rumination frequency (n · day–1), time (min · day–1) and rate (min · g–1 of DM intake) did not differ between treatments ( > 0.05). Apparent total tract digestibility of OM decreased linearly ( = 0.05) with increasing pellet inclusion in the diet. Pelleted cereal-based supplement used in the diet affects feeding behaviour and thus may affect health and welfare of sitatunga in zoological gardens.
Integrin subunits present on human bladder cells displayed heterogeneous func­tional specificity in adhesion to extracellular matrix proteins (ECM). The non-malignant cell line (HCV29) showed significantly higher adhesion efficiency to collagen IV, laminin (LN) and fibronectin (FN) than cancer (T24, Hu456) and v-raf transfected (BC3726) cell lines. Specific antibodies to the a2, a5 and β1 integrin sub- units inhibited adhesion of the non-malignant cells, indicating these integrin partici­pation in the adhesion to ECM proteins. In contrast, adhesion of cancer cells was not inhibited by specific antibodies to the β1 integrin subunit. Antibodies to a3 integrin in­creased adhesion of cancer cells to collagen, LN and FN, but also of the HCV29 line with colagen. It seems that a3 subunit plays a major role in modulation of other integrin receptors especially in cancer cells. Differences in adhesion to ECM proteins between the non-malignant and cancer cell lines in response to Gal and Fuc were not evident, except for the v-raf transfected cell line which showed a distinct about 6-fold increased adhesion to LN on addition of both saccharides. .-Acetylneuraminic acid inhibited adhesion of all cell lines to LN and FN irrespective of their malignancy.
Expression as well as properties of integrins are altered upon transformation. Cell adhesion regulated by integrins is modulated by glycosylation, one of the most frequent biochemical alteration associated with tumorogenesis. Characterisation of carbohydrate moieties of alpha3beta1 integrin on the cultured human bladder carcinoma (T-24, Hu456, HCV 29T) and human normal ureter and bladder epithelium (HCV 29, Hu609) cell lines was carried out after an electrophoresis and blotting, followed by immunochemical identification of alpha3 and beta1 integrin chains and analysis of their carbohydrates moieties using highly specific digoxigenin-labelled lectins. In all the studied cell lines alpha3beta1 integrin was glycosylated although in general each subunit differently. Basic structures recognized in beta1 subunit were tri- or tetraantennary complex type glycans in some cases sialylated (T-24, HCV 29, HCV 29T) and fucosylated (Hu609, HCV 29T). Positive reaction with Phaseolus vulgaris agglutinin and Datura stramonium agglutinin suggesting the presence of beta1-6 branched N-linked oligosaccharides was found in cancerous cell lines (T-24, Hu456) as well as in normal bladder epithelium cells (Hu609). High mannose type glycan was found only in beta1 subunit from Hu456 transitional cell cancer line. On the other hand alpha3 subunit was much less glycosylated except the invasive cancer cell line T-24 where high mannose as well as sialylated tri- or tetraantennary complex type glycans were detected. This observation suggests that changes in glycosylation profile attributed to invasive phenotype are rather associated with alpha3 not beta1 subunit.
 The metastatic transformation of melanocytes is associated with altered expression of adhesion molecules, including αvβ3 and α3β1 integrins. Integrin αvβ3 is a primary vitronectin (VN) receptor, while both integrin types take part in adhesion to VN when they are in complex with uPAR. Although their role in melanoma cell interaction with VN is of great interest, the influence of N-oligosaccharides attached to these glycoproteins is still unappreciated. The present study assesses the role of αvβ3 and α3β1 integrins and the influence of their glycosylation status on WM9 and WM239 metastatic melanoma cell interactions with VN. Cell adhesion to and migration on VN were selected as the studied cell behaviour parameters. Functionblocking antibodies and swainsonine (SW) treatment were used in these tests. Both cell lines interacted with VN in an integrin-mediated but cell-line-specific manner. In WM9 cells, migration was not completely inhibited by antibodies against α3β1 or αvβ3 integrins, suggesting the participation of other VN receptors. In both cell lines in coprecipitation test the formation of an integrins/uPAR complex was shown. In the presence of SW formation of the complex did not occur, suggesting the participation of glycosylation in this proccess. Additionally, the adhesion properties of WM9 cells were changed after SW treatment. Our results suggest that in these two metastatic cell lines integrin-linked N-oligosaccharides influence the VN adhesion receptor activity and function.
The aim of this study was to determine the effect of pelleted cerealbased feed in the diet for addax antelope (Addax nasomaculatus) on feed intake and nutrient digestibility. Four addax females (3 years old) were allocated to 1 of 3 treatments according to 3 × 3 Latin square design and fed 3 diets. Each diet consisted of basal diet and meadow hay. In diet A basal diet consisted of a mixture of ground concentrates, chopped dehydrated lucerne, vegetables and fruits. In diet B 50% and in diet C 100% of dry matter (DM) provided with concentrates and chopped dehydrated lucerne in diet A was replaced with a pelleted cereal-based feed. Meadow hay was offered ad libitum. Two females (sisters) were kept together through the study and thus pen was used as an experimental unit. The diets were formulated to be similar for crude protein and crude fibre content. DM intake (DMI) of basal diet did not differ between treatments (P ≥ 0.52) whereas DMI of hay and total DMI linearly decreased with increasing pellet inclusion in the basal diet (P = 0.05 and P = 0.03, respectively). Digestibility of neutral detergent fibre tended to linearly decrease (P = 0.09) and digestibility of hemicellulose linearly decreased (P = 0.05) with increasing pellet inclusion in the diet. Pelleted cereal-based feed used in the diet for addax had a negative effect on roughage intake and digestibility of structured carbohydrates compared to non-pelleted feed of similar nutrient composition.
The aim of the study was to determine the effect of processing method of cereals used in the diet for nyala antelope (Tragelaphus angasii) on feed intake, eating behaviour, rumination and nutrient digestibility. The study was conducted according to Latin square design (4 × 4) on four adult nyala males. The animals were fed a basal diet consisting of a mixture of cereals (maize, wheat and oat), vegetables and fruits (1.25 kg/day), chopped dehydrated lucerne (0.5 kg/day) and meadow hay (ad libitum). Experimental treatments differed in processing method of cereals offered in the basal diet: 1. finely ground (GR); 2. steam-flaked (SF); or pelleted to obtain: 3. small (6mm diameter size; SP) or 4. large (12mm diameter size; LP) pellet. Feed intake did not differ between the treatments. Eating time of basal diet was longer for GR in comparison to SF and SP (P < 0.01), and also longer for LP in comparison to SP (P = 0.05), with amaximum difference between treatments of 11min/day. Eating rate (g dry matter/min) of basal diet was slower for GR in comparison to SF and SP (P ≤ 0.02). Consumption of meadow hay and chopped lucerne was not affected by cereal processing method. Digestibility of acid detergent fibre was greater (P = 0.05) and digestibility of dry matter and neutral detergent fibre tended (P ≤ 0.08) to be greater for GR than for SP. It can be concluded that, processing method of cereals may affect feeding behaviour of zoo ruminants.
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