Participation of acetylcholine and its receptors in the contractility of inflamed porcine uterus.

This study analyzed the effect of inflammation on acetylcholine (ACh)-induced muscarinic receptors (MR)2 and MR3 conducted contractility of the porcine uterus. On Day 3 of the estrous cycle, either E.coli suspension (E.coli group) or saline (SAL group) was injected into uterine horns or laparotomy was performed (CON group). Eight days later, infected gilts developed severe acute endometritis. Compared to the period before ACh treatment, ACh (10-5 M) increased the tension in myometrium (MYO) and endometrium/myometrium (ENDO/MYO) of the CON group (P < 0.01) and in ENDO/MYO of the SAL group (P < 0.01), the amplitude in strips of the CON (P < 0.05) and SAL (MYO: P < 0.05, ENDO/MYO: P < 0.001) groups and the frequency in strips of the CON (MYO: P < 0.01, ENDO/MYO: P < 0.001) and SAL (P < 0.01) groups. In the E.coli group, ACh (10-5 M) reduced the amplitude in MYO (P < 0.05) and ENDO/MYO (P < 0.001), increased the frequency in MYO (P < 0.01) and ENDO/MYO (P < 0.001) and did not change (P > 0.05) the tension. ACh (10-5 M) in ENDO/MYO of the E.coli group, reduced the tension compared to the CON group (P < 0.05) and the amplitude compared to other groups (P < 0.001), while increased the frequency in relation to the SAL group (P < 0.05). MR2 antagonist (AF-DX 44 116) and ACh (10-5 M) reduced (by 16.92%, P < 0.01) the tension in MYO of the CON group and increased (P < 0.01) it in the E.coli group compared to the period before antagonist and ACh addition. In MYO of the SAL group, the tension was increased (P < 0.01) in response to MR3 antagonist (4-DAMP) and ACh (10-7, 10-6 M). In the E.coli group, these substances did not change (P > 0.05) the tension, but it was lower (P < 0.001) in MYO (ACh: 10-7 M) and ENDO/MYO (ACh: 10-5 M) than in the SAL group. MR2 or MR3 antagonists and ACh (10-5 M) increased (P < 0.05-0.001) the amplitude in strips of the CON and SAL groups and reduced it in the E.coli group (P < 0.001) compared to the period before antagonists and ACh use. This parameter in the E.coli group was lower (P < 0.001) after using MR2 or MR3 antagonists and ACh (10-6, 10-5 M) than in other groups. Both antagonists and ACh (10-5 M) reduced the frequency in the CON, SAL (P < 0.05) and E.coli (MR2 antagonist: P < 0.01, MR3 antagonist: P < 0.05) groups compared to period before antagonists and ACh addition. Data show that ACh reduces the contractility of the inflamed porcine uterus by MR2 and MR3, which suggests that pharmacological modulation of these receptors can be used to raise the contractility of an inflamed uterus.

Innervation of the pineal gland in the Arctic fox (Vulpes lagopus) by nerve fibres immunoreactive to substance P and calcitonin gene-related peptide.

BACKGROUND: The study demonstrates, for the first time, the presence of substance P (SP) and calcitonin gene-related peptide (CGRP) in the nerve fibres supplying the pineal gland in the Arctic fox. MATERIALS AND METHODS: The expression and distribution pattern of the studied substances were examined by double-labelling immunofluorescence technique. RESULTS: The SP-positive fibres enter into the pineal gland through the capsule as the nervi conarii. The fibres formed thick bundles in the capsule and connective tissue septa, from where they penetrated into the pineal parenchyma. Inside the parenchyma, the nerve fibres created basket-like structures surrounding clusters of pinealocytes. The density of intrapineal SP positive fibres was slightly higher in the distal and middle parts of the gland than in the proximal one. Double immunostaining with antibodies against SP and CGRP revealed that the vast majority of SP positive fibres were also CGRP positive. The fibres showing a positive reaction to SP and negative to CGRP were scattered within the whole gland. The fibres immunopositive to CGRP and immunonegative to SP were not observed. In the habenular and posterior commissural areas adjoining to the pineal gland the immunoreactive nerve fibres were not found. Moreover, no immunopositive cell bodies were observed in both the pineal gland and the commissural areas. CONCLUSIONS: These results reveal that SP and CGRP are involved in the innervation of pineal gland in carnivores. In turn we suggest that these peptides can regulate/modulate melatonin secretion.

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.

Long-term testosterone administration affects the number of paracervical ganglion ovary-projecting neurons in sexually mature gilts.

The influence of testosterone (T) overdose on the number and distribution of ovarian neurons in the paracervical ganglion (PCG) in pigs was examined. To identify the ovarian neurons, on day 3 of the estrous cycle, the ovaries of both the control and experimental gilts were injected with retrograde neuronal tracer Fast Blue. From next day to the expected day 20 of the second studied cycle, experimental gilts were injected with T, while control gilts received oil. The PCG was then collected and processed for double-labeling immunofluorescence. T injections increased the T (∼3.5-fold) and estradiol-17ß (∼1.6-fold) levels in the peripheral blood, and reduced the following in the PCG: the total number of Fast Blue-positive neurons, the number of perikarya in the lateral part of the PCG, the numbers of VAChT(+)/SOM(+), VAChT(+)/VIP(+), VAChT(+)/nNOS(+), VAChT(+)/VIP(-), VAChT(+)/DßH(-), VAChT(-)/SOM(-), VAChT(-)/VIP(-), VAChT(-)/nNOS(-) and VAChT(-)/DßH(-) perikarya, In the T-affected PCG, the populations of ovarian perikarya coded VAChT(-)/SOM(+), VAChT(-)/VIP(+) and VAChT(-)/DßH(+), and expressing androgen receptor were increased. After T treatment within the PCG dropped the density of nerve fibers expressing VAChT and/or SOM, VIP, DßH. Obtained data suggest that elevated androgen levels occurring during pathological processes may regulate ovary function(s) by affecting the PCG gonad-supplying neurons.

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.

Calcium concrements in the pineal gland of the Arctic fox (Vulpes lagopus) and their relationship to pinealocytes, glial cells and type I and III collagen fibers.

The aim of the present study was to analyze the presence and morphology of the pineal concretions in the Arctic fox and their relationship to pinealocytes, glial cells and collagen fibers. Pineals collected from 7-8 month-old and 3-4 year-old foxes (6 in each age-group) were investigated. Sections of the glands were stained with HE, Mallory's method and alizarin red S as well as subjected to a combined procedure involving immunofluorescent staining with antibodies against antigen S, glial fibril acid protein (GFAP), type I and III collagen and histochemical staining with alizarin red S. The pineal concretions were found in 2 of 6 investigated Arctic foxes aged 3 years and they were not observed in animals aged 7-8 months. The acervuli were present in the parenchyma and the connective tissue septa. They were more numerous in the distal part than in the proximal part of the gland. The acervuli stained with alizarin red S revealed an intensive red fluorescence, what enabled the use of this compound in a combined histochemical-immunofluorescent procedure. A majority of cells in the fox pineal showed positive staining with antibodies against antigen S, a marker of pinealocytes. GFAP-positive cells were especially numerous in the proximal part of the gland. Both antigen S- and GFAP-positive cells were frequently observed close to the concrements. Collagen fibers of type I and III were found in the capsule, connective tissue septa and vessels. Immunoreactive fibers did not form any capsules or basket-like structures surrounding the concrements.

Pineal concretions in turkey (Meleagris gallopavo) as a result of collagen-mediated calcification.

The intra-pineal calcification is a well-known phenomenon in mammals, however it is almost completely unknown in birds. The aim of the present work was to analyze morphology and genesis of the pineal concretions in the turkey. The studies were performed on the pineals collected from one-year-old turkeys (Meleagris gallopavo). In addition to standard morphological methods, the alizarin red S and potassium pyroantimonate methods were employed for localization of calcium at the light and electron microscopy level. In light microscopy, calcified concretions with diameters from 300 microm to 2 mm and quantities from 3 to 6 per gland were observed in all the examined pineals. They were stained red with alizarin S and showed the presence of collagen in Mallory's staining. Two types of cells were noted inside the concretion: polygonal and elongated ones. Using electron microscopy, three parts were distinguished within the calcification area. The peripheral part contained densely packed collagen fibrils, some elongated cells and numerous pyroantimonate precipitates demonstrating the presence of calcium ions. In the intermediate part, the fibrils were covered by almost continuous sheets of pyroantimonate precipitates and fused side by side. The central part showed an appearance of calcified hard tissue and contained some polygonal (osteocyte-like) cells. The obtained data demonstrated that the formation of the pineal concretions in the turkey is associated with the mineralization of collagen. This process is completely different from the mechanisms responsible for the formation of the concretions in the mammalian pineal.

Healthy diet in primary care: views of general practitioners and nurses from Europe.

BACKGROUND: Most of the national colleges of general practitioners (GPs) do not have their own dietary/nutritional tools, and GPs and nurses do not have the time, knowledge, or skills to advise their patients about desirable dietary practices. OBJECTIVE: To assess the usefulness of a simple and practical guide on healthy diet to be used by European GPs and nurses. DESIGN: A postal survey was mailed to 171 GPs and nurses from 12 European countries to obtain information about the usefulness of a guide on healthy diet developed by EUROPREV. RESULTS: The perception of health professionals is that the main source of information on healthy diet for the population was the media. In all, 95% of GPs and nurses reported that the guide was useful; 93, 95, and 82% reported that the concepts were concise, easy to understand, and realistic, respectively. Also, 77% reported that the type of counselling recommended was feasible and could be applied, 94% reported that the implementation measures proposed could be effective and 88% reported that the Traditional Mediterranean Diet Pyramid is useful, but some concerns about the content were mentioned. CONCLUSIONS: GPs and nurses from Europe think that a practical guide on healthy diet developed by EUROPREV could be used to advise patients in primary care, although the Traditional Mediterranean Diet Pyramid should be modified.