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The aim of the study was to estimate the size of bone marrow-origin stem/progenitor population in 2-year old nonpregnant Holstein-Friesian heifers. Quantitative and qualitative analysis was done using scanning cytometry and confocal microscopy of mammary tissue slices labelled with the combination of two markers: Sca-1 (marker of stem-progenitor cells) and CD45 (marker of hematopoietic cells). The average (±SEM) percentage of Sca-1POS CD45POS cells was 0.89 ± 0.21. They were localized mainly outside of mammary ducts, in the stroma and sometimes intraluminally. Our results indicate that the subpopulation of Sca-1POS cells bearing CD45 antigen may enrich the niche of mammary stem/progenitor cells from the bone marrow and participate in the growth of the mammary gland in post-pubertal heifers.
The cj0183 and cj0588 genes identified in the Campylobacter jejuni NCTC 11168 genome encode proteins with homology to virulence factors found in other bacteria. Previous studies showed that single mutation in the cj0183 gene does not affect adhesion of C. jejuni to the Caco-2 cell line whereas protein encoded by cj0588 is involved in adherence to the Caco-2 cells. In the presented study differences in invasion index were observed between mutants in both genes and single mutation of cj0588 in 81116 and 81-176 C. jejuni strains. This fact indicates that Cj0183 protein might play some role in invasion of bacteria into host cells.
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Control of development of gastrointestinal system in neonates

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Our recent studies of structure and function of gastrointestinal tract mucosa revealed that the domestification of Sus scrofa corresponds with the significant slowing of the organ development. On top of genetic potential, the nutritional factors (or more precisely - lack of certain biologically active substances in the feed of pregnant sows) are responsible. Moreover, feeding neonates with milk replacers instead of mother's milk further slows down the development. This is manifested by reduced mitotic activity in the crypts and enhanced apoptosis of enterocytes. The negative effects consist of slower replacement of fetal type, vacuolated enterocytes to adult type enterocytes, modified profile of brush border enzymes, alterations in intestinal mucosa barrier, higher susceptibility to infectious agents, and many others. On the other hand, farmers in order to intensify the production, shorten the suckling period imposing the neonatal piglets to be weaned at 3-4 weeks of life and even earlier. Altogether, it makes the weaning disorders one of the most important problems in pig husbandry, and the mortality of piglets in the leading pig-producing countries still reaches 10%. A number of strategies have been developed to counteract the post-weaning problems. One of them is to stimulate the development of the gastrointestinal tract of the neonate by supplementation of the sow diet with certain biologically active substances and plants. The other idea is to speed up the postnatal development of the gut mucosa for example by plant lectins. Lessons from pig studies can be also useful in human nutrition and medicine since the development of porcine gastrointestinal tract shows a great similarity to that of humans.
Recent decades have brought rapid development in the field of nanotechnology, which has led to applications of nanoparticles in many industries. Unique properties of nanoparticles and their biocompatibility increase their potential as drug carriers in drug-delivery systems. Prof. Marek Godlewski’s team from the Institute of Physics PAS has developed wide band-gap metal oxide nanoparticles doped with rare-earth metals for applications as fluorescent markers. The potential of those nanoparticles to cross the closed gut barrier after alimentary application has prompted their use in drug delivery systems. In this study, we show that after conjugation with a model bioactive substance, lectin (Phaseolus vulgaris), these nanoparticles retained their advantageous properties and, following oral administration (10 mg/ml in RO, 0.3 ml/mouse), entered a variety of organs in the mouse model. Internal organs collected at key time points were analysed under a scanning cytometer and a confocal microscope. The results show that the conjugation reduced, but did not completely abolish, the capacity of nanoparticles to penetrate physiological barriers (intestinal, blood-brain barrier) in the organism.
The use of soybean in human and animal nutrition is limited because of high content of bioactive compounds: enzyme inhibitors, polyphenols, goitrogens, phytates, saponins, sugars, and agglutinins. The damage of intestinal mucosa structure was previously observed in animals fed soybean supplemented diets. Hence, the objectives of the presented study were to compare intensity of epithelium remodeling processes in different intestinal segments, and to evaluate the influence of the 1% of soybean dietary supplementation on the processes in intestinal mucosa. The experiment was performed on 30 Wistar rats fed AIN-93 based diets. Animals were divided randomly into three groups: control (CTRL), with 1% of raw soybean (RS) and with 1% of soaked and boiled soybean (BS). The samples of: duodenum (DUO), proximal jejunum (PROX), mid-jejunum (MID), distal-jejunum (DIST) and ileum (ILE) were collected. The following processes in these samples were evaluated: mitosis (Ki-67), apoptosis (Cpp32), autophagy (MAP I LC3) and DNA damage (p53). Present data show that modification of soybean by soaking and subsequent boiling markedly influences the enterocyte turnover in the small intestine mucosa. Increased mitotic ratio in the intestine of rats fed with boiled soybean masks the negative effects of soybean on the small intestine structure.
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Into the unknown - the death pathways in the neonatal gut epithelium

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Apoptosis is a fundamental process in the development of the fast growing intestinal mucosa. Apoptotic cells are present along the whole length of the villi and in the crypts. The mechanisms involved in the induction of apoptosis in the gut mucosa are still unknown. Cytokines are believed to play a role in auto- and paracrine models because the cells are dying in so-called "packets" containing neighboring cells. In the rapidly developing gut of neonates, the apoptosis rate is transiently reduced in the first days of life, enhancing the growth of mucosa. Afterwards, apoptosis plays a role in the exchange of the enterocyte population, facilitating maturation of the mucosa. The presence of autophagic cells has been confirmed for the first time in the developing gut. Deprivation of growth factors during feeding artificial milk formula led to an increased apoptosis rate. Supplementation with leptin reduced cell apoptosis and increased the mitosis-to-apoptosis ratio. Autophagy was also diminished. The key to healthy gut mucosa growth in early life, especially in fast-growing animals, is colostrum, which supplies nutritional and defensive components together with supplementary growth factors, cytokines and hormones essential for growth and maturation of gut mucosa.
Luminescent nanocrystals and quantum dots have great potential for use as fluorescent markers in biology and medicine. However, their first generations were based on the heavy-metal core, which was unstable and shed heavy-metal ions into biological media. This, coupled with a lack of information on their biodistribution and pharmacokinetics, rendered them unusable for purposes outside research. The recently developed nonheavy metal nanocrystals are a promising material for future medical use. Yet, the controversies over their application, absorption and biodistribution remain. Various recent papers present different results on the uptake of nanocrystals and on their intracellular and organ distribution. In our study, we focused on the question of how the size and shape of nanocrystals affect their duodenal absorption after intragastric gavage (IG) and distribution to the liver. Commercial bulk nanoparticles and hydrothermal nanoparticles produced at the Institute of Physics PAS were the same in composition and excitation-emission range, but significantly different in shape and size. Adult mice (n = 24) aged 3-6 months were kept in standard living conditions (12 h day-night cycle), fed ad libitum with unobstructed access to water. Following a 1-week adaptation period, an RO water suspension of nanoparticles (50 µg/ml) was administered by IG. No changes in the behaviour of the mice or pathophysiological changes in their organs were observed following IG. The control group received an identical volume of RO water by IG. Cross-sections of the organs were examined both qualitatively and quantitatively by confocal microscopy and scanning cytometry. Following IG, both types of nanoparticles entered the duodenum in a similar time, but only the smaller, elongated hydrothermal nanoparticles were absorbed through the intestinal epithelium and distributed throughout internal organs (p ≤ 0.001). In conclusion, we found that the size and shape of nanocrystals is crucial for their bioavailability.
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Large-conductance K+ channel openers induce death of human glioma cells

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Large-conductance Ca2+-activated K+ channels (BKCa channels) are highly expressed in human glioma cells. It has been reported that BKCa channels are present in the inner mitochondrial membrane of the human glioma cell line LN229. In the present study we investigated whether BKCa-channel openers, such as CGS7181 (ethyl 2-hydroxy-1-[[(4-methylphenyl)amino]oxo]-6-trifluoromethyl-1H-indole-3-carboxylate) and CGS7184 (ethyl 1-[[(4-chlorophenyl) amino]oxo]-2-hydroxy-6-trifluoromethyl-1H-indole-3-carboxylate), affect the functioning of LN229 glioma cell mitochondria in situ. In the micromolar concentration range CGS7181 and CGS7184 induced glioma cell death. Morphological and cytometric analyses confirmed that both substances trigger the glioma cell death. This effect was not inhibited by the pan-caspase inhibitor z-VAD-fmk. Lack of DNA laddering, PARP cleavage, and caspase 3 activation suggested that glioma cell death was not of the apoptotic type. We examined the effect of CGS7184 on mitochondrial membrane potential and mitochondrial respiration. Potassium channel opener CGS7184 increased cell respiration and induced mitochondrial membrane depolarization. The latter was dependent on the presence of Ca2+ in the external medium. It was shown that CGS7184 induced an increase of cytosolic Ca2+ concentration due to endoplasmic reticulum store depletion. In conclusion, our results show that CGS7181 and CGS7184 induce glioma cell death by increasing the cytosolic calcium concentration followed by activation of calpains.
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