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The immunoreactivity of haemagglutinin (HA) polypeptides of equine influenza virus was compared among the strains isolated in Poland, using H3 monoclonal antibody. A stronger signal in immunoblot reaction was observed for A/equi/Pulawy/2008 HA polypeptides compared to A/equi/Pulawy/2006, despite the fact that both strains are phylogenetically closely related and belong to Florida clade 2 of American lineage. The strongest signal, observed in the case of A/equi/Pulawy/2008, seemed to be connected with the presence of G135, 1213, E379, and/or V530 instead of R135, M213, G379, and 1530 present in A/equi/Pulawy/2006 HA sequence. This implies that point mutations within amino acid sequences of HA polypeptides of equine influenza virus may change their immunoreactivity even when they are not located within five basic antigenic sites.
The vasoactive intestinal peptide (VIP) and opioid family member Leu⁵-enkephalin (LENK) have already been established as playing independently significant roles in the functioning of the female genital tract. However, the mutual influence of both neuropeptides on female genital function has not been examined until now. Therefore, the aim of this study was to compare the distribution of VIP- and/or LENK-immunoreactive (IR) structures throughout the female genital tract of the pig. Immunohistochemical examination revealed that the great majority of the immunopositive structures co-expressed both peptides. Nevertheless, a small population of exclusively VIP- or LENK-IR processes and perikarya were also distinguished. The muscular layer of the organs examined revealed the greatest density of VIP- and/or LENK-IR nerve fibers. The mucosa of the ampulla, isthmus, cervix and vagina was supplied with a moderate number of single labeled LENK-IR processes, while exclusively VIP-IR fibers were found mainly in vaginal mucosa. The infundibulum was found to be poorly supplied with single labeled VIP- or LENK-IR fibers. The paracervical ganglion (PCG), the expected source of VIP- and/or LENK-IR nerve fibers innervating the organs under investigation, has been found to contain double labeled LENK-/VIP-IR as well as single labeled VIP-IR perikarya. The great number of specific co-localization between VIP and LENK in nerve processes of the porcine female genital organs may indicate a functional regulatory interaction between the neuropeptides studied, requiring further study.
In our study we used c-Fos protein to identify whether cells containing calretinin (CR) in the rat piriform cortex are engaged in the response to stress stimulation and to find out how this expression changes during maturation (PC). The material consisted of Wistar strain rats of between 0 and 120 days of age divided into 9 groups. Each group consisted of 5 experimental and 3 control rats. Animals from the experimental groups were exposed to the open field test throughout 10 minutes. The control animals were kept in a home cage. In all age-related control rats weak c-Fos immunoreactivity was observed. Our results showed that cells containing c-Fos following an acute open field test were observed predominantly in layers II and III of the PC just after birth. Their number then increased and stabilised on P30. We had already observed immature CR-ir cells at birth. In the 4th week of life these neurons achieved maturity. Their number increased to P90 and decreased in older animals. CR-ir neurons were localised mainly in layer II and to a lesser degree in layers III and I of the PC. Double immunostaining c-Fos/CR revealed that the level of co-localisation was low. Only small differences were observed between the anterior and posterior parts of the PC. In the anterior part a higher number of CR-ir neurons was found. The difference in the level of co-localisation between the anterior and posterior parts was age-related and differentiated. Our results may suggest that during maturation CR-ir neurons of the piriform cortex are not the main population engaged in response to the open field test.
Choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT) are currently accepted specific markers for cholinergic neurons. Current knowledge concerning the organization of the hypothalamic cholinergic system has been derived predominantly from laboratory animals. In the present study, applying immunocytochemistry the authors investigated the morphology and distribution of the ChAT- and VAChT-immunoreactive nerve cells and terminals in the porcine hypothalamic preoptic, supraoptic and tuberal nuclei. ChAT-immunoreactive perikarya were present in the tuberal arcuate nucleus. Numerous VAChT- -positive terminals were found in the external layer of the median eminence, while perikarya occupied the perifornical, dorso-caudal and dorsal hypothalamic nuclei. These results provide morphological indications that the cholinergic hypothalamic system may affect the secretory function of the median eminence in pigs.
Glycation (non-enzymatic glycosylation) is a spontaneous reaction that occurs during food processing, storage and preparation of food, which can have a significant influence on physiochemical and biological properties of food proteins. Glycation has been used mostly for improvement of functional properties of proteins although there is increasing concern of possible changes in biologic properties of glycation products. The influence of dry-heat glycation with glucose on immunoreactive properties of wheat salt-soluble proteins has been studied. It has been shown that glycation caused differences in electrophoretic patterns (both 1D and 2D), as well as in Western-immunoblotting with rabbit IgG and human IgE. A significant increase in immunoreactivity has been observed, as well as a decrease in free amino group. Thermostable, immunoreactive and susceptible for glycation protein band with molecular weight of approximately 13 kDa (by SDS-PAGE) has been selected for the N-terminal sequence analysis and determined to be an alpha-amylase inhibitor. Dry heat glycation is a very potent but nondestructive method of protein glycation, that leads to high degree of substitution and changes in both physicochemical and biological (immunological) properties of food proteins.
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