Trichinella spiralis: the influence of short chain fatty acids on the proliferation of lymphocytes, the goblet cell count and apoptosis in the mouse intestine.

This study was carried out to determine the influence of short chain fatty acids (SCFA) on spleen and mesenteric lymph node lymphocyte proliferation, goblet cells and apoptosis in the mouse small intestine during invasion by Trichinella spiralis. BALB/c mice were infected with 250 larvae of T. spiralis. An SCFA water solution containing acetic, propionic and butyric acids (30:15:20 mM) was administered orally starting 5 days before infection and ending 20 days post infection (dpi). Fragments of the jejunum were collected by dissection 7 and 10 dpi, and were examined for apoptotic cells in the lamina propria of the intestinal mucosa, and for goblet cells. The proliferation index of the cultured spleen and mesenteric lymph node lymphocytes with MTT test was also determined. The orally administered SCFA solution decreased the proliferation of mesenteric lymph node lymphocytes in the mice infected with T. spiralis at both examination times, but did not influence the proliferative activity of the spleen cells. Seven dpi, both in the spleen and mesenteric lymph nodes, the highest proliferation index of concanavalin A (Con A)-stimulated lymphocytes was found in the group of uninfected animals receiving SCFA animals. This tendency could still be seen 10 dpi in the mesenteric lymph nodes but not in the spleen, where the proliferation index in this group had significantly decreased. In vitro studies revealed, that butyric and propionic acids added to the cell cultures suppressed the proliferation of Con A-stimulated mesenteric lymph nodes and spleen lymphocytes taken from uninfected and T. spiralis-infected mice. Acetic acid stimulated proliferation of splenocytes taken from uninfected mice but did not affect lymphocyte proliferation in mesenteric lymph nodes from uninfected or infected mice. Orally administered SCFA increased the number of goblet cells found in the epithelium of the jejunum 7 dpi, but this number had decreased 10 dpi. The number of apoptotic cells in the lamina propria of the intestinal mucosa of animals infected with the T. spiralis and receiving SCFA was also lower, particularly 10 dpi. The above results show that SCFA can participate in the immune response during the course of trichinellosis in mice.

Influence of diet type on the inclusion of plant origin active substances on morphological and histochemical characteristics of the stomach and jejunum walls in chicken.

Three hundred and thirty-six 1-day-old male Hubbard HI-Ye broiler hybrids, kept in battery cages, were fed with diets based on maize (groups I and II) or wheat and barley (groups III and IV) and supplemented with or without plant extract (XT* 100 mg/kg) containing 5% carvacrol, 3% cinnamaldehyde and 2% of capsicum oleoresin. The morphological and histochemical examinations were carried out on days 21 and 42 of bird's age. The middle part of glandular part of the stomach and 30 mm long segment from the central part of the small intestine (jejunum) were taken out and then prepared for morphometrical and histochemical assays. Mobilization of mucocytes in superficial epithelium of the glandular stomach and increased secretion of neutral mucopolysaccharides and small amounts of sialomucins with or without local cell disruption with releasing of large amounts of mucus were observed in both 'grain' groups of 21-day-old birds fed with extract. In some animals, particularly those fed mixtures with plant extract, the folds of the proventriculum mucosa were fused into large, unshaped structures. In groups fed with plant extract the mucus secretion intensity and accumulation inside cells of the gastrointestinal mucosa were slightly higher. Morphological changes on gastrointestinal mucosa observed in young chickens fed XT were reduced in older animals. The results of this study showed that the increased releasing of large amounts of mucus and the creation of a thick layer of mucus on glandular stomach and wall of jejunum in chickens fed diets with plant extract could suggest villi-related protective properties of the use of the carvacrol, cinnamaldehyde and capsaicin mixture. This can explain the reduced possibility of adhesion to epithelium and number of Escherichia coli, Clostridium perfringes and fungi in the intestinal content of bird fed with XT supplemented diet. In morphometrical parameters of depth of jejunum crypt and height of villi, the influence of kind of grain and extract supplementation was observed in 21-day-old chickens only. The significant interaction between higher jejunum wall villi layer was observed only in chickens fed on maize diet supplemented with plant extract.

The phenomenon of apoptosis in the course of experimental trichinellosis in mice.

Mice infected with 200 Trichinella spiralis larvae were killed at 3-69 days post infection (dpi) and the jejunum and masseter muscles were sectioned in a cryostat and examined in the Tunell method with "In situ Cell Detected Kid POD" of Boehringer-Mannheim. Data concering the exact localization and dynamic of apoptic cells in both organs are presented. The authors conclude that apoptosis plays a important role in the pathogenesis of trichinellosis.

Macrophages during infection with Trichinella spiralis in mice.

Behaviour of macrophages in experimental mice trichinellosis was investigated using the immunoenzymatic technique with monoclonal antibodies CD11b/CD18 within the framework of avidin-biotin-DAB. The maximum and earliest mobilization of macrophages 7 day post infection (dpi) was observed in the lamina propria of the intestinal mucosa. The highest level of macrophages in the muscles tissue was noted on day 21 of infection, however as early as in 28 dpi, their maximum level was observed inside the larval capsules. They line, especially between 35-42 dpi internal capsule surface.

The cells observed inside capsules of larvae in the course of experimental trichinellosis in mice.

In the course of trichinellosis in mice the numerous cells inside larval capsules were observed. Beginning on 14 day post infection (dpi) they were seen in about 7% of infected muscles fibers but at 21 dpi the percentage of capsules with the cells amounted to 77. In the next stage of infection the number of capsules containing cells continued to increase and reach almost 100% at 60 dpi. The identification of the cells was carried out by the indirect immunoenzymatic method using anti-CD4+, anti-CD8+ and anti-macrophage (Mac-1 antigen) monoclonal antibodies. In course of observation the level of CD4+ cells decreased from 11.3 to 0.7, of CD8+ from 17.7 to 3.0 and of macrophages from 71.0 to 23.1%. The most numerous CD4+ cells were seen at 14 dpi while the CD8+ and macrophages at 21 dpi. Beginning on 28 dpi negatively reacting cells inside larval capsules were also observed. The number of these cells increased from 23.8 to 73.2% at 60 dpi and their identification requires further investigations. The role and the function of all these cells are discussed.

[The influence of ricin on the course of experimental infections of mice with Trichinella spiralis]. / Wplyw ricinu na przebieg doswiadczalnego zarazenia myszy nicieniami Trichinella spiralis.

Mice received 5 ng of ricin 24 hours after infection (experiment I) or 1 ng of ricin twice, 24 hours and 17 days of infection (experiment II). Animals were killed in 7, 14, 21, 28, 35, 42 and 60 days after infection. In the jejunum and masseter muscle sections, CD4+ and CD8+ lymphocytes, mast cells and eosinophils were studied. Heavy suppression of CD8+ lymphocytes, strong eosinophils and less pronounced mast cells stimulation was observed in the jejunum of mice received ricin (experiment I). Worm expulsion in intestine was faster than in the control (opposite results in experiment II). The composition of cells infiltration in the muscle was in both experiments similar to the control, however, fewer CD4+ lymphocytes were observed in larva capsule and there were fewer muscle larvae. Therefore CD8+ cells are believed to take part in restricting only the muscle stage of trichinellosis.

[The influence of phytohemagglutinin P on CD4+ and CD8+ cells in the course of experimental trichinellosis in mice]. / Wplyw fitohemaglutyniny P na zachowanie sie komórek CD4+ i CD8+ w przebiegu eksperymentalnej wlosnicy u myszy.

Experiments were carried out with mice of CFW inbred strain infected with 200 Trichinella spiralis larvae and divided into two groups: experimental and control. The experimental mice were injected with phytohaemagglutinin P (PHA-P), 24 h before infection. The all animals were killed between 7-133 days post infection and the pieces of spleens, mesenteric lymph nodes, jejunum and masseter muscles were sectioned in a cryostat and examined with monoclonal antibodies (anti-Thy 1, 2, anti-CD4+ and anti-CD8+) by immunoenzymatic method. In the animals treated with PHA-P, more numerous CD4+ cells were observed in comparison with the control. The CD8+ cells were stimulated only in the muscles. The most important problem were the CD4+ and CD8+ cells inside the larvae capsules. The role of the CD4+ and CD8+ cells in the immunopathological changes in the course of trichinellosis was discussed.

CD4+ and CD8+ cells in mice infected with Trichinella spiralis and treated with cyclosporine A.

The mice infected with 200 Trichinella spiralis larvae were injected intraperitoneally with Cyclosporine A (CyA) between 14-18 days post infection (dpi). The drug was administered in a dosis of 50 mg/kg/day. The animals were killed at 21, 28, 35, 42 and 60 dpi and the fragments of spleen, mesenteric lymph node, jejunum and musculus masseter were sectioned in a cryostat and fixed in acetone. The slides were examined with monoclonal sera by the immunofluorescent or immunoenzymatic method. It was found that the number of CD4+ cells in the control and in the CyA-treated mice was similar but in the animals receiving the drug the reaction was less intensive. The stimulation of CD8+ cells of CyA treated mice--especially in the jejunum--was stronger than in the control animals. This fact is important because the CD8+ cells are the APC cells in this organ.

The influence of adriamycin on the CD4+ and CD8+ cells in the course of trichinellosis in mice.

The effect of adriamycin on the behaviour of CD4+ and CD8+ cells in the course of trichinellosis in mice has been studied. The animals infected with 200 larvae per mouse were administered intraperitoneally adriamycin (Adriblastin from Farmitalia) at 1st and 28th day post infection (dpi), in a dosis of 0.2 mg. The mice were killed weakly for 6 weeks and then at the 60th dpi. The examinations were made on histologic sections from the spleen, mesenteric lymph node, jejunum and masseter muscle using immunofluorescent and immunoenzymatic methods with monoclonal antibody. The mice receiving adriamycin exhibited more CD8+ cells in the intestinal mucosa and by the end of the experiment also in the muscles in comparison with the control animals, which, however, did not affect the course of the infection.

[Effect of cyclosporin A on the course of the muscular phase of trichinellosis in mice]. / Wplyw cyklosporyny A na przebieg miesniowej fazy wlosnicy u myszy.

The authors tried to determine the role of TH lymphocytes in the course of muscular trichinellosis in mice. The investigations were carried out on the CFW mice infected with 200 Trichinella spiralis larvae and treated with Cyclosporine A to suppress TH1 lymphocytes. In animals treated with the drug more numerous cells inside the larval capsules and more numerous granulomas compared to control were seen. At the same time the mean number of larvae was lower (69.2%) than in control. The results show that TH1 lymphocytes do not play the significant role in the destruction of muscular larvae. This is the first conclusion and requires of course further investigations.

Effect of a single dosis of 96% ethanol on rat gastric mucosa. Part II. Histoenzymological studies.

The histoenzymological studies of ATPase and SDH enzymes in rats' gastric mucosa after a single 90 s administration of 96% ethanol proved considerable changes in the enzymes activity in course of injury and subsequent regeneration of mucosa.

[Mucopolysaccharides of the gastric mucosa in rats after ethanol-induced injury]. / Mukopolisacharydy blony sluzowej zoladka szczurów w przebiegu uszkodzenia etanolem.

The experiment was performed in 34 Buffalo rats. After laparotomy 2 ml of 96% ethanol was injected into the stomach for 90 sec. At 15 min, 1 hour, 24 hours and 96 hours mucopolysaccharides of the mucosa of bottom parts of the stomach were analysed histochemically. A direct reaction to concentrated alcohol was massive exfoliation and disintegration of the surface epithelium or abrupt release of the mucus from the superficial epithelial cells, pits and neck leading to a formation of a thick muco-cellular layer of gel precipitating reepithelization. Apart from an increase in the amount of surface mucus significant changes in its composition were found. They included an increase in the amount of sulfo- and sialomucins affecting the level of mucous viscosity. It was also shown that AB+ mucous cells of the neck were, a good marker of proliferative zone; state of the latter affects markedly the rate of regeneration of the damaged gastric mucosa.

[Differences in the reaction of the mucosa of the gastric fundus and pylorus to high concentration of ethanol]. / Zróznicowanie reakcji blony sluzowej czesci dennej i odzwiernikowej zoladka szczurów na dzialanie stezonego etanolu.

Histopathological morphometric analysis were made to determine the degree and extent of the stomach mucosa lesions in fundus and antrum of 36 rats after 15 min., 1, 24 and 96 hours after ethanol insult. At the early stage the intensity of the lesion caused by direct influence of ethanol was in both parts similar. Evident differences occurred after 1 hour, and especially after 24 hours, and were caused by secondary, endogenous agents. Among them the most important seems to be the specificity of the vascular net reaction to alcohol stimulus in each part of the stomach, differences in secretion intensity and composition of the superficial mucosal gel, as well as in the contents of some active substances.

[Pathomorphological picture of various internal organs of rats after long-term exposure to toxic substances in raw coke gas]. / Obraz patomorfologiczny niektórych narzadów wewnetrznych szczurów poddanych dlugotrwalemu dzialaniu substancji toksycznych zawartych w surowym gazie koksowniczym.

The authors carried out morphological evaluation of a long-term action of toxic substances at low concentrations (about 1 NDS), contained in raw coke oven gas, i.e. gases: CO, H2S, SO2, NH3 and benzene, as well as dusts containing: C, SiO2, Pb and Cd on the liver and kidneys of three-month old male rats of the Wistar strain. The animals were exposed to the above toxic substances in an experimental chamber for 24 hr, and for 8 hr every day during 12 and 18 months. It was found that pathogenic action of the substances contained in raw coke oven gas was not the total action of all chemical components, but their complicated resultant. Significant differences in the intensity of morphological changes in lungs were observed between rats exposed for 8 hr ans 24 hr/day, in favour of the former. Such a relationship was not observed between 12 and 18 month exposition. Besides the catarrh of the air passages and cellular-fibrous changes in pulmonary alveoli, focal metaplasia of cells of bronchial epithelium was observed, which suggested the possibility of carcinogenic action of coke oven gases and dusts. No distinct morphological differences were found in the liver and kidneys between 8 and 24 hr exposition, whereas great differences were found between 12 and 18 months exposition. Chronic interstitial pyelitis was observed in kidneys after 18 month exposition.