The influence of selenium and vitamin E supplementation on cytological assessment of red blood cell line of bone marrow in fallow deer kept in captivity.

Cytological evaluation of bone marrow smears stained by May-Grünwald Giemsa method was performed. The smears came from 20 fallow deer (Dama dama) 3 days old divided into 2 groups each consisting of 10 animals. The experimental group (E) received intramuscularly selenium and vitamin E at a dose of 3.0 ml (tocopherol acetate - 50 mg, sodium selenite - 0.5 mg, solvent - 1 ml) in the 3rd day of age. The control group (C) did not receive any supplementation or placebo. For hematological analyzes blood was collected three times: on 0, 15th and 25th day of the experiment. Serum concentration of selenium and vitamin E was determined using high perfor- mance liquid chromatography and glutathione peroxidase activity (GSH-Px) by kinetic method. On the 15th day after supplementation, a statistically significant increase in the percentage of erythroblastic cell line was observed in bone marrow smears. At that time, the increase in GSH-Px activity in the E group was also observed, reaching the value of 165.3 U/gHb, which was statisti- cally significant. The percentage of proerythroblasts (8.23% in group E and 5.02% in group C) differed significantly between groups at the 25th day after supplementation. This study revealed that supplementation of selenium and vitamin E resulted in an increase in the number of erythro- cytes to an average of 13.5 (˟ 10¹²/l) in the experimental group on 25th day with a significant increase in hemoglobin to 193 g/l in the experimental group.

Influence of simvastatin on red blood cell line in porcine bone marrow.

Simvastatin is a drug commonly used to reduce the cholesterol level. 32 clinically healthy female pigs with a bodyweight of 35-40 kg, kept in standard laboratory conditions were chosen for the experiment and divided into 2 groups (control and experimental) consisting of 16 animals. The experimental group received simvastatin orally at a dose of 40 mg (one tablet once a day) for 56 days, and at the same time the control group received placebo (empty gelatin capsules). Bone marrow smears and peripheral blood samples were evaluated. The obtained results indicate that simvastatin may inhibit erythropoiesis even after a relatively short period of administration, and observed changes can be the cause of some symptoms (for example anemia) occurring during statin therapy.

Distribution and chemical coding patterns of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons in the enteric nervous system of the porcine stomach cardia.

The aim of this study was to determine the presence of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and co-localisation of CART with vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (n-NOS), vasoactive intestinal polypeptide (VIP), substance P (SP) and leu-enkephalin (LENK) in the enteric nervous system of the porcine gastric cardia by using a double-labelling immunofluorescence technique. CART-LI neurons were observed in the myenteric plexus (18.2±2.6%). A dense network of CART-LI nerve fibers was mainly observed in the muscular layer. CART showed co-localization mainly with VAChT, n-NOS, VIP and to a lesser degree with LENK and SP. Distribution of CART and its co-localization with other neurotransmitters suggest that this peptide plays an important role in gastric motility in the pig.

Zearalenone-induced changes in the lymphoid tissue and mucosal nerve fibers in the porcine ileum.

This is the first study to examine zearalenone-(ZEN) induced changes in the immune system of the ileum and substance P-(SP-) and vasoactive intestinal peptide-(VIP-) immunoreactive nerve fibers in the mucosa, which participate in the regulation of intestinal functions under physiological conditions and during pathological processes. The aim of this study was also to identify potential relationships between selected immune and neural elements in ileal Peyer's patches in pigs that were and were not exposed to ZEN. The experiment was performed on 10 prepubertal gilts divided into two groups: the experimental group (n=5) where ZEN was administered at 0.1 mg kg-1 feed day-1 for 42 days, and the control group (n=5) which was administered a placebo. The tissue levels of cytokines were determined by enzyme-linked immunosorbent assay which revealed elevated concentrations of IL-12/23 40p and IL-1 ß in animals exposed to ZEN. Flow cytometry revealed a lower percentage of CD21+ lymphocytes in pigs exposed to ZEN in comparison with control animals. The tissue levels of neuropeptides were evaluated in the dot blot procedure which demonstrated higher concentrations of VIP and SP in experimental pigs. In experimental animals, numerous VIP-like immunoreactive processes were observed, and SP-immunoreactive nerve fibers formed a very dense network. Our results demonstrate for the first time that ZEN can modify the chemical coding of nerve structures in the gastrointestinal system. Those modifications can be attributed to ZEN's impact on estrogen receptors or its pro-inflammatory properties, and they reflect changes that take place in the nervous system at the transcriptional, translational and metabolic level.

Immunohistochemical evidence of the co-localisation of cocaine and amphetamine regulatory peptide with neuronal isoform of nitric oxide synthase, vasoactive intestinal peptide and galanin within the circular muscle layer of the human caecum.

The enteric nervous system consists of about one hundred million of neurons. In big mammals (including humans) intestinal enteric neuronal cells are grouped into three types of intramural ganglia located within myenteric, as well as outer and inner submucosal plexuses, which are connected by numerous nerve fibres. Both nerve fibres and cell bodies located in the gastrointestinal tract utilise a broad spectrum of active substances. One of them is cocaine- and amphetamine-regulated transcript peptide (CART). The goal of the current study was to determinate the distribution and degree of co-localisation of CART with substances taking part in intestinal motor activity by double labelling immunofluorescence technique. During the study CART-, neuronal isoform of nitric oxide synthase (nNOS)-, vasoactive intestinal peptide (VIP)- and/or galanin (GAL) - like immunoreactive (LI) nerve fibres in the circular muscle layer of the human caecum were observed in all patients studied. The degree of co-localisation of particular substances with CART depended on their type. The majority of CART-LI fibres contained simultaneously nNOS, slightly lower degree of co-localisation was observed in the case of the VIP, while simultaneously CART- and GAL-positive nerve fibres were observed less often.

A population of nesfatin 1-like immunoreactive (LI) cells in the mucosal layer of the canine digestive tract.

The aim of this study was to investigate for the first time nesfatin-1-like immunoreactive (LI) cells in the mucosal layer of the canine digestive tract. Distribution of nesfatin-1 - LI cell bodies and the co-localization of nesfatin-1 with somatostatin (SOM), cocaine-and amphetamine-regulated transcript peptide (CART), protein gene product 9.5 (PGP 9.5) and vasoactive intestinal peptide (VIP) were studied by immunohistochemistry in the selected parts of the canine digestive tract. Evaluation of the nesfatin 1-LI cells number has been based on the counting of cell bodies per observation field. Nesfatin-1-LI cells amounted to 0.87±0.06, 1.06±0.02, 0.85±0.11 and 0.56±0.13 in gastric fundus, duodenum, jejunum and descending colon, respectively. The co-localization of nesfatin-1 with the other substances studied was not observed. Functions of nesfatin-1 - LI cells in the canine digestive tract are unknown.

The relationship and co-localization of vasoactive intestinal peptide (VIP)- and Leu5-enkephalin (LENK)-immunoreactivity in the female genital tract of the pig.

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.

Neither axotomy nor target-tissue inflammation changes the NOS- or VIP-synthesis rate in distal bowel-projecting neurons of the porcine inferior mesenteric ganglion (IMG).

The present study was aimed at disclosing axotomy- and inflammation-induced changes in the chemical coding of retrogradely labelled distal bowel-projecting neurons in the porcine IMG. Particular attention was paid to the changes in the expression pattern of vasoactive intestinal polypeptide and nitric oxide synthase (as a marker of nitric oxide) in affected cells, as these substances are thought to play a crucial role in the regeneration of injured sympathetic neurons. However, while both pathological processes failed to induce an increase in the number of sympathetic bowel-projecting neurons exhibiting vasoactive intestinal polypeptide or nitric oxide synthase, axotomy, but not target-tissue inflammation, led to the upregulation in the expression pattern of galanin, pituitary adenylate cyclase-activating peptide and/or Leu5-enkephalin in the affected perikarya. On the other hand, axotomy resulted in a diminished density of vasoactive intestinal polypeptide-immunoreactive intraganglionic nerve fibres, whilst target-tissue inflammation evoked a distinct increase in the number of visible vasoactive intestinal polypeptide-immunoreactive terminals, especially in those regions where bowel-projecting neurons were located. Thus, the data obtained in the present study run counter to the results of the injury-related responses observed in neurons of the sympathetic chain ganglia, suggesting the existence of either species- or target tissue-dependent differences in the injury-induced responses of the affected sympathetic neurons.

Distribution of growth associated protein (GAP-43) immunoreactivity in nerve fibers supplying the pig pineal gland.

An immunohistochemical study of the pig pineal gland was carried out using monoclonal mouse antiserum against growth-associated protein GAP-43. The pineal glands were obtained from the 3, 5, 8 weeks old piglets. The immunopositive nerve fibers were observed in the pineal gland as well as in the habenular and the posterior comissural areas. They formed a dense network in the habenular area and the proximal part of the pineal gland. In the comissural area and in the apical part of the gland. single positive fibers were observed. The obtained results may suggest a difference in the plasticity of innervation between the particular regions of the pineal gland.