Tripeptidyl peptidase I activity in porcine lumbar spinal ganglia - a histochemical study.

Distribution of tripeptidyl peptidase I (TPPI) activity in the structures of porcine lumbar spinal ganglia (LSG) was studied by enzyme histochemistry on cryostat sections from all the ganglia using the substrate glycyl-L-prolyl-L-methionyl-5-chloro-1-anthraquinonyl hydrazide (GPM-CAH) and 4-nitrobenzaldehyde (NBA) as visualization factor. Light microscopic observations showed TPPI activity in almost all the LSG structures. The enzyme reaction in different cell types was compared semi-quantitatively. Strong reaction was observed in the small neurons, satellite ganglia cells and some nerve fibers. Weak reactivity was found in the large sensory somatic neurons, whereas moderate reaction for TPPI was determined in the middle sensory somatic neurons and some nerve fibers. Statistical analysis by one-way ANOVA showed no significance of difference (when p⟨0.05) for the number of TPPI positive neurons per mm2. The original data obtained by the enzyme histochemistry method give us a reason to presume that TPPI actively participates in the functions of all the neuronal structures in porcine LSG. According to our results, it could be suggested that TPPI activity is important for the functions of autonomic and somatic sensory neurons.

Histochemical Study of Heparin-positive Mast Cells in the Terminal Part of Porcine Ductus Choledochus and Papilla Duodeni Major.

The study presented in detail the localization and density of mast cells (MCs) in the intramural part of the common bile duct (CBD) and in the major duodenal papilla (MDP) of domestic swine. MCs' density (number/mm(2) ) in different layers of both of the duct and papilla was evaluated after toluidine blue staining. Their number was higher in the lamina propria mucosae than in the tunica muscularis of the studied structures. The localization of berberine-positive, (heparin containing) MCs and the ratio between them and toluidine blue-positive MCs with γ-ma metachromasia was also established. Ratios of heparin-containing MCs in comparison with all toluidine blue-positive MCs were found as follows: ductus choledochus - 32% in the subglandular connective tissue of lamina propria mucosae in the intramural part of the duct; m. sphincter ductus choledochus - 31% in the circular and 0.06% in the longitudinal muscle layer; subserosa - 59%; papilla duodeni major - 0.03% in the subepithelial connective tissue and 34% in the subglandular connective tissue of lamina propria mucosae, respectively. The established large difference in heparin-positive MCs in both the subepithelial and subglandular connective tissues of CBD and MDP, respectively, is an evidence for the existence of mucosal and connective tissue MCs.

S-100 protein-positive mast cells in canine paranal sinus (Sinus paranalis).

The aim of this study was to establish the immunocytochemical expression of S-100 protein in mast cells, localized in the wall of dog's paranal sinus. Control serial sections were used for immunocytochemical detection of tryptase-positive mast cells. It was observed that S-100-positive cells have the same morphology and localization as the tryptase-positive mast cells, which indicated that S-100-positive cells are most probably mast cells with abilities as dendritic cells. In conclusion, for the first time, the current study gave evidence that mast cells in this organ possess one more function, such as dendritic cells.

NADPH-d-positive mast cells in the canine paranal sinus.

This study aimed to investigate the enzyme histochemical expression of NADPH-d in mast cells in the wall of the paranal sinus in male and female dogs. NADPH-d-positive cells with weak, medium and strong enzyme histochemical expression were observed in the stroma of the sinus near the blood vessels of the microcirculatory bed and around the apocrine and sebaceous glands. In the same areas, mast cells with similar dimensions and morphology were demonstrated by metachromasia on paraffin and cryostat cross-sections and stained with 0.1% toluidine blue in McIlvane's buffer (pH 3). These findings suggest that the mast cells that are located in the stroma near the blood vessels, the lining epithelium and the glands correspond with the cells with marked NADPH-d activity. The possibility of mast cells having nitric oxide activity could be used in the regulation of mast cells function when treating paranal sinus tumours and inflammations.

NADPH-d cells (mast cells) around and within the autonomic nerves of porcine renal hilus.

The enzymehistochemical localization of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) reactive cells next and within the autonomic nerves in the porcine kidney's hilus, close to renal artery and vein as well as in its adventitia, was studied. It was established that the prevailing part of NADPH-d-reactive cells were distributed next to and within NADPH-d-reactive autonomic fibers around the renal vein. Comparatively small number of reactive cells was found around the renal artery, but single NADPH-d-reactive cells were found in the neighbouring autonomic nerves as well. The cells observed next to nerves had much more expressed reaction (granules) than those, located in the nerves. Toluidine blue staining on frozen and paraffine sections from the same areas showed well-expressed gamma-ma metachromasia in mast cells with similar or identical localization. This indicated that observed reactive cells were mast cells. The obtained data convincingly showed that porcine NADPH-d cells may produce nitric oxide and in this way most probably they were involved in the function of autonomic nerves in the renal blood vessels (vasodilatation) and the kidney function, as well.

Immunohistochemical study of 3 beta-hydroxysteroid dehydrogenase in dog's perianal sinus.

In this study, the localization of 3 beta-hydroxysteroid dehydrogenase (3beta-HSD) activity in the wall of canine perianal sinus (PS) was determined. The 3 beta-HSD activity was found out both in the cytoplasm of cells, situated in the propria and forming clusters adjacently to apocrine glands and in the cytoplasm of some epithelial cells in apocrine cells' glands. The results obtained about the 3 beta-HSD activity allowed us to propose a role of this enzyme in PS development and possibly, in tumourogenesis.

Endothelin-positive mast cells in porcine renal artery and vein.

For a first time the endothelin (ET)-positive mast cells were examined in the wall of kidney renal artery and vein. The specimen's were collected from six 8-month-old Danmark Landrace pigs, immediately after slaughtering. Mast cells immunopositive to ET granules were observed in the wall of both artery and vein. In the renal artery, they were found mostly between the media and the adventitia. Some mast cells were found in the media, next to smooth muscle cells. Relatively few mast cells were found in the intima and between intima and tunica media. In the renal vein a smaller number of mast cells were observed. They showed similar localization as in the renal artery. Immunopositive mast cells were established also close to endothelial cells - mostly between internal elastic membrane and basal membrane of the endothelium. In conclusion, on the basis of obtained results, presumptions for active participation of ET (most probably mainly ET-1) in the motility of the vessels' smooth muscle and for stimulation of nitric oxide release from the intimal endothelial cells were made.

Histological changes of the feline cervix, endometrium and placenta after mid-gestational termination of pregnancy with aglepristone.

This study was conducted to investigate endometrial and placental structural changes that occurred in response to mid-gestational termination of pregnancy in queens using aglepristone, a progesterone receptor antagonist. Thirteen European Shorthair pregnant queens were either treated with aglepristone (10 mg/kg body weight; subcutaneously) twice on days 25 and 26 after first mating (am; group I; n = 9), or remained untreated and served as control (group II; n = 4). Queens of group I were ovariohysterectomized between days 30 and 41 am, either at the onset (n = 3) or during (n = 1) abortion and 12 h (n = 1), 24 h (n = 3) or 10 days after abortion (n = 1). Queens of group II were ovariohysterectomized on day 30 am. Tissue was collected from the cervix, and the interplacental zone as well as the paraplacenta/placental girdle of the uterus, subjected to standard histological procedures and evaluated using light microscopy. During abortion, gaps appeared within the paraplacenta and the placental girdle which were filled with blood, leading to an embryo-maternal disconnection. Blood was also observed within the uterine lumen as well as the interstitial mucosal stroma of the cervix and the placental girdle zone and probably originated from damaged venules, whilst arterioles remained intact. As the interval between abortion and surgery increased, the interstitial and luminal haemorrhages became less pronounced and completely disappeared except interstitial remnants 10 days after abortion. The endometrial regeneration was not fully completed on day 10 after abortion and a few cystically dilated glands were evident. In conclusion, abortion of queens through aglepristone given during mid-gestation is assumed to be the result of damage of uterine venules. This leads to an interstitial haemorrhages and bleeding into the uterine lumen, subsequently resulting in utero-placental detachment.

Morphological and immunocytochemical investigations on mast cells in porcine ureter.

Morphological, morphometric, histochemical and immunocytochemical investigations on mast cells, located in the wall of ureter of 8 months aged pigs were performed. Mast cells were found in all three layers of ureteral wall, but their distribution was irregular and the number unequal. It was established that alcian blue (AB)-positive mast cells were significantly more than toluidine blue (TB)-positive mast cells. A statistically significant smaller number of both AB and TB-stained mast cells were observed in the tunica mucosa. The largest number of mast cells was found in the tunica muscularis. In the adventitia, mast cells were higher in number in the main connective tissue than in the connective tissue near the blood vessels. Mast cells stained with TB showed variably expressed gamma-metachromasia, which was best visible in those situated in the lamina propria of the mucosa. The prevailing parts of mast cells, however, were AB-positive after AB-safranin staining. This was mostly found in mast cells of the tunica muscularis and in mast cells of perivascular location in the tunica adventitia. Immunocytochemically, mast cells were found to be positive for histamine and vasoactive intestinal polypeptide in the muscle coat, and to histamine in the adventitia, as well. On the basis of obtained results it was presumed that the mast cells in porcine ureter most probably took part not only in keeping of local homeostasis, but played also an important role of mobility of smooth muscle cells in the middle layer of ureter on one hand, and, on the other, in the adventitial blood vessels.

Structural examination of tryptase-, and VIP-positive mast cells in the common bile duct of patients with lithiasis.

The morphology of tryptase-, and vasoactive intestinal polypeptide (VIP)-positive mast cells was examined immunohistochemically in 38 common bile ducts collected from patients with secondary chronic cholangitis and varying degrees of inflammatory activity. Mast cells numbers in chronic exacerbated and chronic sclerotic cholangitis were significantly higher as compared with those in controls (72.4 cells/mm2 and 25.2 cells/mm2 vs. 5.9 cells/mm2; p < 0.0001, Student's t test). The increased number of tryptase-positive mast cells in chronic exacerbated cholangitis correlated with the severeness of inflammatory infiltration. In cases of chronic exacerbated cholangitis, the increased number of mast cells was detected in conjunction with active fibroplasia. In chronic sclerotic cholangitis mast cells were lower in number as compared with exacerbated cholangitis and were observed in relation with inactive fibrosis. Numerous VIP-positive mast cells were found in all patients with cholangitis. Ultrastructural immunocytochemistry showed tryptase positivity to be localized over either electron-dense or particulate granules with a mean diameter of 0.261+/-0.073 microm or 0.171+/-0.053 microm, respectively. VIP positivity was formed as a finely or coarsely granular pattern over larger electron-dense granules of 0.475+/-0.14 microm in diameter. Tryptase-positive mast cells were located mainly in and around surface and glandular epithelium. The involvement of tryptase- and VIP-positive mast cells in inflammation, fibrosis and epithelial reactions in the common bile duct is discussed.

Morphological study of mast cell localization in the wall of the proximal tubule in the domestic swine kidney.

Morphological studies to localize mast cells in the wall of the convoluted and straight parts of the proximal tubules of the domestic swine kidney were performed. Kidney perfusion was carried out in 8-month-old pigs and semi-thin and ultrathin sections were prepared after a routine treatment. A light microscopic study showed that mast cells were not frequently found. Most were localized in the wall of the proximal convoluted tubule, while in the proximal straight tubule wall they were fewer in number. Most of the mast cells were oval in shape, their size being typical for this species, and the granules were reddish in colour. Three types of granule containing an evenly distributed granular-fibrous substance were found by TEM examination: the first type had a single electron-optically dense belt under the membrane; the second showed optically empty spaces between the substance and the cell membrane; the granules from the third type had two parallel dense belts below the granular membrane and were the smallest in size. Some of the granules of the first type were connected with the granular endoplasmic reticulum. A very narrow intercellular area and single desmosome-like junctions were found between the mast cells and the epithelial cells. Protrusions of plasmalemma and granules outside the cells were not established. On the basis of these results and data from the scientific literature, a comparative analysis was carried out. This showed certain differences between the results obtained and previous findings on the morphology of domestic swine mast cells and granules.

Morphologic peculiarities of the renal cortical vasculature connected with blood redistribution in the kidney of the domestic pig.

The cortical blood vessels from 2 to 8 month old Camborow hybrid pigs and Belgium landrace pigs were studied using histologic and semithin sections as well as corrosion casts and cleared preparations of intravascular injection. It was established that the cortical blood vessels of the porcine kidney possess peculiarities that may regulate the redistribution of blood in the cortex. These peculiarities were found in the interlobular arteries, in the efferent arterioles of midcortical nephrons, in afferent arterioles of juxtamedullar glomeruli, and in the glomerular capillary networks. It was established that the number of renal corpuscles is different in the various zones of the cortex.

Peculiarities of the connection between mesangial and glomerular endothelial cells in the domestic-swine kidney.

The peculiarities of the connection between mesangial and endothelial cells are investigated in material from non-perfused swine kidneys. It is established that cytoplasm fragments of the endothelial cells, located between the mesangial angles, abruptly extend their size, and borders between them are difficult to discern. They possess single microvilli on their lumenal surface, some of which are highly corrugated, branched and fenestrated. In those parts, the mesangial-cell excrescences are tightly enveloped by the endothelium, some of them contacting directly with the capillary lumen. On the border between the mesangium and the juxtamesangial part of the endothelium, two types of vesicles, directed towards the capillary lumen, are observed. The first type of vesicle is bigger and has a deeply narrowing section, which is directed towards the mesangium. The second type of vesicle is enveloped by a two-layer membrane, which, in almost all cases, is connected to the endothelium. Peculiarities in the localization of the podocyte excrescences are also detected, which gives reason to presume that the epithelial cells play an important part in the biomechanics of the renal glomerulus.

Methods for permanent conservation of biological objects and anatomic preparations in synthetic resins.

Two methods for preparing packets of synthetic resin for permanent preservation of biological objects and anatomical tissue have been developed. The first method is applied when adding corrosive preparations to epoxide resin. The corrosive preparation is soaked in a mixture of epoxide resin and hardener in a ratio, experimentally determined. The induration proceed at room temperature for 12 h. The second method has two options and is applied for the conservation of biological objects and anatomical preparations in polymethylmethacrylate by a successive immersing of the object in a system of dehydrating mixtures: 50%, 70%, 90% and 96% water solution of methanol and ethanol and in a mixture of acetone and ethanol (1:1) and in acetone or in methylene chloride. The object, thus prepared, is impregnated in vacuum with polymer of methylmethacrylate, benzoyl peroxide and stearic acid or in a composition containing: methylmethacrylate, benzoyl peroxide, dibutylphthalate and tertiary aromatic amine, in optimum correlations of the components, experimentally determined. The induration proceed by heating in a water bath (37 + 3 degrees C) for 3 h and at 30 degrees C temperature in the course of 6-8 h in the first option or at room temperature for about 3 days in the second option. The advantages of the proposed methods are: the use of cheap and available materials for the production of a simplified technological scheme of strongly transparent packages, preserved over a long period; preserving the sizes, the form and natural colour of the objects and preparations as well as a possibility for selective tinting of the separate tissues and systems in the preserved biological objects and anatomical preparations; a possibility for stereomicroscopic observation of the objects and preparations preserved.

Microscopical and ultrastructural studies of the renal artery in domestic swine.

Both renal arteries from six, 8-month old, Camborow hybrid pigs were examined in histological, semithin and ultrathin sections. It was established that the membrana elastica interna showed an expressed degree of wrinkliness. In certain sections, sometimes occupying 20% of the diameter of across-section, its curves were considerably higher and wider than those in the remaining part. The inner elastic membrane in these sections was cleaved in two layers. Estates of lengthwise-situated smooth muscle cells of an elliptical form were located in the middle shell (tunica media). Furthermore, a great quantity of mast cells were found in the middle shell. The electron-microscopic observations showed that their granules had various sizes and shapes. The outer shell (tunica adventitia) had an irregular thickness. In some parts, a few elastic fibres entered the middle shell. They had a tuft-like shape with a wedge-like or elongated configuration, and were situated among the layers of smooth-muscle cells.

Renal venous architectonics in domestic swine.

Renal venous architectonics were investigated in 240 kidneys from 120 swines at an age of 1-8 months. The methods used were: preparation, corrosion and X-ray examination. It was established that, in 75% of the cases, v. renalis cranialis and v. renalis caudalis were present. In the rest of the cases (25%), v. renalis intermedia was present. It was also observed that right and left renal veins formed distentions, which enveloped the caudal medial part of each adrenal gland. V. adrenalis (suprarenalis) was not formed as an independent blood vessel. Venous blood from the adrenal gland drains through 2-3 orifices in these distentions. This study describes vv. prelobares, vv, interlobares and vv. arcuatae. Anastomoses were found between them. On that basis this concluded that the renal venous system in domestic swine is not segmented.

Morphological studies of the valves of the kidney vein in domestic swine.

It was established macroscopically that 37.5% of the renal veins investigated possess valves. Valves were observed on both sides, but predominantly on the left side of the kidneys (14.58%). Some valves were also observed in the main branches of the renal veins. The structure of the valves was studied with the use of light and electron microscopy. The most important finding was the observation of mast cells in the valves. This is probably a common biological phenomenon, since mast cells have been observed in the cardiac valves and the wall of the renal artery and vein.

[Arterial vessels of the kidney in domestic swine]. / Arterialni sudove na bubreka pri domashnata svinia.

By histological preparation, measuring, corrosion and angiography were studied the arterial blood vessels of 120 kidneys from 60 individuals at different ages. It was established that the kidney artery in the most cases (70%) divides by the dichotomous type in cranial and caudal or in dorsal and ventral branches, and rarely (30%) by the disseminated type in 3 branches--two cranial and one caudal. Investigated were the position, course and branching of the prelobar, interlobar, the arcuate, the inter- and intra-lobular arteries.