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.

Effect of a single-dose parenteral selenium supplement administered to pregnant dairy cows on selenium and iron concentrations and immune status of calves.

The study was performed on 21 H-F calves divided into 3 groups of 7 animals each. The first group was composed of calves whose mothers did not receive an injection of Se preparation. The second and third groups consisted of calves whose mothers were administered injections of Se and vitamin E in a single dose of 10 ml and 30 ml, 10 days before the expected parturition date. 24 hours after birth, blood samples were collected from all calves to determine Se, Fe and IgG concentrations and the activity of GSH-Px and GGT. The results of the study indicate that the administration of a single-dose Se supplement to cows in late pregnancy increases Se concentration in calves and promotes passive transfer of immunity from the mother to offspring.

Immunohistochemical characteristics of porcine intrahepatic nerves under physiological conditions and after bisphenol A administration.

BACKGROUND: The neurochemistry of hepatic nerve fibres was investigated in large animal models after dietary exposure to the endocrine disrupting compound known as bisphenol A (BPA). MATERIALS AND METHODS: Antibodies against neuronal peptides were used to study changes in hepatic nerve fibres after exposure to BPA at varying concentrations using standard immunofluorescence techniques. The neuropeptides investigated were substance P (SP), galanin (GAL), pituitary adenylate cyclase activating polypeptide (PACAP), calcitonin gene regulated peptide (CGRP) and cocaine and amphetamine regulated transcript (CART). Immunoreactive nerve fibres were counted in multiple sections of the liver and among multiple animals at varying exposure levels. The data was pooled and presented as mean ± standard error of the mean. RESULTS: It was found that all of the nerve fibres investigated showed upregulation of these neural markers after BPA exposure, even at exposure levels currently considered to be safe. These results show very dramatic increases in nerve fibres containing the above-mentioned neuropeptides and the altered neurochemical levels may be causing a range of pathophysiological states if the trend of over-expression is extrapolated to developing humans. CONCLUSIONS: This may have serious implications for children and young adults who are exposed to this very common plastic polymer, if the same trends are occurring in humans.

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.

Neuropeptide profile changes in sensory neurones after partial prepyloric resection in pigs.

This report details the first identification of the sources of sensory innervation of the porcine stomach prepyloric region. The Fast Blue (FB) retrograde tracing technique detected the sensory prepyloric neurons in the bilateral nodose ganglia (NGs) as well as thoracic dorsal root ganglia (DRGs). Double-labelling immunofluorescence demonstrated expression of substance P (SP), calcitonin gene-related peptide (CGRP), neuronal isoform of nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP) and galanin (GAL) in both NGs and DRGs. Additionally, we found that partial resection of the stomach prepyloric area increased expression of the SP, CGRP, NOS, VIP and GAL in the prepyloric sensory neurons. In the control left NGs, both a higher total number of FB-positive perikarya as well as a higher percentage of the peptides expressing prepyloric neurons were visualized than in the right NGs. However, compared to the control group, prepyloric resection evoked greater increases in peptide expression in the right-side NGs sensory neurons. In the ganglia of this side, the proportion of the SP-IR perikarya increased by approximately 15%, while CGRP-IR increased by 28%, NOS-IR 14%, VIP-IR 43% and GAL-IR 13%. On the opposite left side, the ganglia proportion of the CGRP-IR perikarya increased by approximately 10%, while NOS-IR increased by 3%, VIP-IR 36% and GAL-IR by 2%. The only decrease (by 5%) was observed in the case of SP expression. We also found that 92% of the sensory neurons originated from NGs and 8% from DRGs. Our results indicate that, in the pig, SP, CGRP, NOS, VIP and GAL participate in the vagal sensory transduction from the stomach prepyloric area. Moreover, increased expression of the peptides and neuronal isoform of nitric oxide synthase in the sensory neurons following transection of their peripheral dendrites suggests their possible participation in the neuronal recovery and/or reinnervation process.

Acetylsalicylic acid-induced changes in the chemical coding of extrinsic sensory neurons supplying the prepyloric area of the porcine stomach.

Acetylsalicylic acid is a popular drug that is commonly used to treat fever and inflammation, but which can also negativity affect the mucosal layer of the stomach, although knowledge concerning its influence on gastric innervation is very scarce. Thus, the aim of the present study was to study the influence of prolonged acetylsalicylic acid supplementation on the extrinsic primary sensory neurons supplying the porcine stomach prepyloric region. Fast Blue (FB) was injected into the above-mentioned region of the stomach. Acetylsalicylic acid was then given orally to the experimental gilts from the seventh day after FB injection to the 27th day of the experiment. After euthanasia, the nodose ganglia (NG) and dorsal root ganglia (DRG) were collected. Sections of these ganglia were processed for routine double-labelling immunofluorescence technique for substance P (SP), calcitonine gene related peptide (CGRP), galanin (GAL), neuronal isoform of nitric oxide synthase (nNOS) and vasoactive intestinal polypeptide (VIP). Under physiological conditions within the nodose ganglia, the percentage of the FB-labeled neurons immunoreactive to particular substances ranged between 17.9 ± 2.7% (VIP-like immunoreactive (LI) neurons in the right NG) and 60.4 ± 1.7% (SP-LI cells within the left NG). Acetylsalicylic acid supplementation caused a considerable increase in the expression of all active substances studied within both left and right NG and the percentage of neurons positive to particular substances fluctuated from 47.2 ± 3.6% (GAL-LI neurons in the right NG) to 67.2 ± 2.0% (cells immunoreactive to SP in the left NG). All studied substances were also observed in DRG neurons supplying the prepyloric region of the stomach, but the number of immunoreactive neurons was too small to conduct a statistical analysis. The obtained results show that ASA may influence chemical coding of the sensory neurons supplying the porcine stomach, but the exact mechanisms of this action still remain unknown.

Co-expression of PACAP with VIP, SP and CGRP in the porcine nodose ganglion sensory neurons.

Our previous study revealed the expression of substance P (SP) and calcitonin gene-related peptide (CGRP) in sensory distal ganglion of the vagus (nodose ganglion) neurons in the pig. As these neuropeptides may be involved in nociception, the goal of these investigations was to determine possible expression of vasoactive intestinal polypeptide (VIP), SP and CGRP in the pituitary adenylate cyclase-activating polypeptide-immunoreactive (PACAP-IR) porcine nodose perikarya. Co-expression of these substances was examined using a double-labelling immunofluorescence technique. To reveal the ganglionic cell bodies, the pan-neuronal marker protein gene product 9.5 (PGP 9.5) was used. Quantitative analysis of the neurons revealed that 67.25% of the PGP 9.5+ somata in the right-side ganglion and 66.5% in the left side, respectively, co-expressed PACAP-IR. Moreover, 60.6% of the PACAP-IR cells in the right-side ganglion and 62.1% in the left, respectively, co-expressed VIP. SP-IR was observed in 52.2 and 39.9% of the right and left ganglia, respectively. CGRP was found in 27.7 and 34.1% of the right and left distal ganglion of the vagus, respectively. High level of co-expression of PACAP with VIP, SP and CGRP in the distal ganglia of the vagus sensory perikarya directly implicates studied peptides in their functional interaction during nociceptive vagal transduction.

Reduction of the number of neurones in the caudal mesenteric ganglion innervating the ovary in sexually mature gilts following testosterone administration.

The effect of testosterone on the morphological and chemical plasticity of the porcine caudal mesenteric ganglion (CaMG) ovary-projecting neurones was investigated. To identify the neurones on day 3 of the oestrous cycle, the ovaries of both the control and experimental gilts were injected with Fast Blue retrograde neuronal tracer. From next day until day 20 of the anticipated second studied cycle, experimental gilts were injected with testosterone, whereas control gilts received oil. Testosterone injections increased testosterone (by approximately 3.5-fold) and 17ß-oestradiol (by approximately 1.6-fold) levels in the peripheral blood and decreased the following in the CaMG: the total number of Fast Blue-positive perikarya (including small ones); the population of small perikarya in the caudal, ventral and dorsal ganglional regions; the numbers of dopamine-ß-hydroxylase (DßH) and/or neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL) small and large perikarya; the numbers of small perikarya containing DßH (but not NPY, SOM, GAL); and the density of DßH and/or NPY, SOM nerve fibres. A disappearance of small and large non-noradrenergic perikarya and an increase in the total number of androgen receptor-immunoreactive perikarya was noted. Our results suggest that elevated androgen levels occurring during pathological states may regulate ovary function(s) by affecting the CaMG gonad-supplying neurones.

Co-localisation of cocaine- and amphetamine-regulated transcript peptide and vasoactive intestinal polypeptide in the myenteric plexus of the porcine transverse colon.

Cocaine- and amphetamine-regulated transcript peptide (CART) is a substance,which can play the role of neuromediator and/or neuromodulator in nerve structures within the gastrointestinal tract. However knowledge concerning its functions and co-localisation with other neuronal factors is rather scarce. During the present investigation the co-localisation of CART and vasoactive intestinal polypeptide (VIP) in the neurons of meyenteric plexus within the porcine transverse colon was studied using double immunofluorescence technique and semiquantitative arbitrary scale of the frequency of presence CART+/VIP+, CART+/VIP- and CART-/VIP+ neuronal cells. The most often (+++) CART-/VIP+ neurons were encountered, neurons simultaneously immunoreactive to CART and VIP were observed somewhat rarer (++) and only single (+) CART+/VIP- perikarya were visible. The present study reports for the first time on the co-localisation of CART and VIP in myenteric neurons of the porcine transverse colon.