Ján Jakubovský

Metastasizing Adenocarcinoma of the Female Prostate (Skene's Paraurethral Glands): Histological and Immunohistochemical Prostate Markers Studies and First Ultrastructural Observation

The case of a 46-year-old women with well-differentiated adenocarcinoma of the female prostate (Skenes paraurethral glands and ducts) with inguinal metastases is reported. Besides adenocarcinomatous structures, also more solid parts of the tumor and anaplastic regions with dark cells were found on histological examination. Clear cancerous cells were typical for glandular and solid tumor parts. The cancerous cells showed distinct immunohistochemical positivity of prostate specific antigen (PSA) and prostate (specific) acid phosphatase [P(S)AcP]. These are the first published results of electron microscopic examination of formalin fixed tissue showing the ultrastructure of female prostate carcinoma, comparable to that of the male prostate carcinoma. In the female, similar to the male, the prostate carcinoma probably originates from the secretory (luminal) cells of the female prostatic glands.

Endotoxin-induced aggravation of preservation-reperfusion injury of rat liver and its modulation

BACKGROUND/AIMS In clinical transplantation, exposure of donors to gut-derived endotoxin occurs frequently and may adversely affect liver transplantation therapy. The aim of this study was to investigate: 1) whether brief exposure of rats to endotoxin before liver procurement aggravates the early phase of reperfusion injury of hepatic explants; and if so 2) whether Kupffer cell activation is a contributing factor to liver injury; and 3) whether heparin and pentoxifylline could minimize this effect. METHODS Male Wistar rats were injected with 0.2-4.0 mg/kg of Escherichia coli lipopolysaccharide 2 h prior to liver harvest. After preservation in University of Wisconsin cold-storage solution, the livers were reperfused using a blood-free perfusion model. To inactivate Kupffer cells, some rats were pretreated with gadolinium chloride or liposome-encapsulated dichloromethylene-diphosphonate before lipopolysaccharide administration. The other rats received lipopolysaccharide with heparin or pentoxifylline. RESULTS In a dose-independent fashion, lipopolysaccharide impaired portal flow during graft reperfusion. In a dose-dependent way, lipopolysaccharide increased lactate dehydrogenase release into the perfusate and decreased bile flow and bromosulfophthalein excretion. Gadolinium chloride, liposomal dichloromethylene-diphosphonate, heparin, and pentoxifylline reduced lactate dehydrogenase release by 34%, 43%, 59%, and 64%, respectively, and improved functional parameters of the liver. A 52-fold increased neutrophil infiltration in the liver sinusoids after lipopolysaccharide exposure was not affected significantly by the drugs studied; however, heparin reduced markedly neutrophil activation. CONCLUSIONS The results of this investigation provide direct evidence that aggravation of preservation-reperfusion injury of rat liver by endotoxin is mediated by Kupffer cell-dependent mechanism(s) and it can be minimized by heparin and pentoxifylline.

Fluorescence of hematoxylin and eosin-stained histological sections of the human spleen

A major problem in the morphometric evaluation of human spleen is the simple but reliable determination of the border between T-cell and B-cell dependent areas, and other structures of the spleen. It was investigated whether cryostat sections of frozen surgical specimens of the human spleen and sections of paraffin-embedded specimens could be used for this purpose after being stained with haematoxylin and eosin and mounted in autofluorescence-free medium for fluorescence microscopical evaluation. Comparison was made with sections that were immunohistochemically-stained for fibronectin and collagens type II and type IV. Both in cryostat sections and paraffin sections, fluorescence was found in circumferential reticulum of periarterial lymphatic sheets, arterial terminals, arterial walls and walls of red pulp sinuses in the spleen. Evaluation was hindered by fluorescence of erythrocytes in paraffin sections but not in cryostat sections. Results were similar as those obtained with immunohistochemical fibronectin staining and are sufficient for morphometric evaluation or orientation in the tissue in case of neoplasia.

Collagen type IV in epithelial tumours of colon.

Collagen type IV in the lamina propria mucosae is one of the main components of the basement membrane of normal and transitional colon mucosa. The aim of the present study was to assess the use of anti-collagen type IV antibodies in the evaluation of biological activity of epithelial tumours of the colon. Formalin-fixed and paraffin-embedded specimens of polyps and carcinomas of the colon from 14 patients were analyzed. In transitional mucosa around epithelial tumours, only minor deformities of the evenly thick collagen type IV-containing basement membranes were found. This pattern was different in polyps where collagen type IV-positive basement membrane components extended between basolateral membranes of epithelial cells. Local changes of collagen type IV positivity in basement membranes of polyps were observed. Positivity of epithelial basement membranes disappeared in adenocarcinomas but there was an increased positivity in fibrillar components of stroma. Basement membranes of microvessels in lamina propria mucosae were also positive for collagen type IV. Similar observations were made in the stroma of polyps. Our results indicate that loss of collagen type IV in basement membranes of adenocarcinomas is related to loss of differentiation and the malignant potential of epithelial tumours of colon.

Rhythmic changes of human female uroepithelial squamous cells during menstrual cycle. Transmission and scanning electron microscopic study.

The cellular component of the fluid of female urethral expulsions was analysed in two healthy women (37-year-old multipara and 27-year-old nullipara) on days 1, 10, 12, 14, 26 and 27 of their menstrual cycle. Transmission and scanning electron microscopy were used to determine standard values of uroepithelial squamous cells of the fluid of female urethral expulsions on different days of the menstrual cycle. In agreement with the known changes of these cells found in cytological smears during the cycle, electron microscopy confirmed characteristic changes in the appearance of the fine cell structure and the space configuration of the squamous cells. Changes in the desmosomal junctions among squamous cells during the cycle were found. Striking adherence of bacteria and mucosubstances to the squamous cell surfaces were observed in the first half of the menstrual cycle and at the time of ovulation.

Influence of starving on the rat gastric mucosa — Lightand electron microscopical findings —

The histology and ultrastructure of the rat gastric mucosa were investigated during 168 hours of starvation. An increased desquamation of individual foveolar cells was found. In the preserved cells of the foveolae, the content of the PAS positive mucosubstances did not change during starvation, and no changes took place in the appearance and in the amount of the mucous granules at the electron microscopic investigation. The number of lipid droplets increased in the mucous foveolar cells within 24 and 48 hours. During starvation the mitochondria (mainly in the parietal cells) were enlarged and contained rare mitochondrial cristae. Some mitochondria were distintegrated and removed by lysosomes. The number of lysosomes (mainly cytosergresomes) was markedly increased n parietal cells. A collapse of the intracellular canaliculi occurred as well as a narrowing in the tubulovesicular profiles. In chief cells the profiles of the granular endoplasmic reticulum and Golgi apparatus were reduced. It was shown that the ultrastructural changes induced by starvation can be interpreted functionally in changed histochemical parameters of the gastric mucosa.

Ultrastructure of the human antral G cells during fasting.

The ultrastructure of the human antral G cells was examined in gastrobiopsies of 7 healthy volunteers fasting 24 to 240 hours. In contrast to the animals gastric C cells during fasting, the dissolution of the substance of gastrin granules (but not emiocytosis of granules) was observed in the human antral G cells during long fasting. The authors consider the possibility of artificially inducing the dissolution of the substance of the granules only the technique of gastrobiopsy itself.

A novel way of liver preservation improves rat liver viability upon reperfusion.

Background/aimCurrently, the liver is cold-preserved at 0∼4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion.MethodsIn the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A: 45 min and Group B: 24 h) or by slow cooling HTK solution (from 13 °C to 3 °C) during the initial 45 min of preservation (Group C: 24 h). In the second set of experiments, additional groups of livers were evaluated: Group BB—preservation according to Group B and Group CC—preservation according to Group C. Further, some livers were preserved at 13 °C for 24 h. Livers were then reperfused using a blood-free perfusion model.ResultsBile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2∼3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 °C for 24 h exhibited severe ischemic injury.ConclusionThese results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.

Gastroprotective effect of pentacaine: Role of mast cells

Pentacaine was found to prevent the development of acute haemorrhagic lesions induced by ethanol in rats in a dose-dependent way. Electron microscopy in the untreated group showed extensive disruption of the surface epithelium and deep necrosis of the mucosa after ethanol exposure. Degranulation or even complete destruction of mast cells was observed. The microvasculature exhibited several signs of derangement. After pentacaine treatment, these signs were absent and no degranulation of mucosal mast cells was observed. The mast cell-mediated effect of pentacaine appears to be only one component of its gastroprotective action.

A Comparative Study of the Morphology of the Systemic Anaphylactic Reaction (SAR) and the Shock Reaction Induced by Antigen-Antibody Complexes in Rabbits

Differences in the morphological picture of the systemic anaphylactic reaction (SAR), and of the shock reaction induced by antigen-antibody (ag-ab) complexes in rabbits, are described. In rabbits SAR is characterized by acute distension of the lungs, with oedematous swelling in the vicinity of bronchi and veins, intravascular stasis of basophils without degranulation or visible fixation of the inducing antigen, small clusters of blood platelets, leukostasis, minimal decomposition of neutrophil granulocytes and absence of circulating or fixed ag-ab complexes. The shock reaction induced by ag-ab complexes is characterized by acute distension of the lungs without bronchospasm, swelling of the vicinity of bronchi and veins, hyaline thrombi particularly in the pulmonary microcirculation, leukostasis, formation of antigen-antibody complexes and thrombocyte clusters in the microcirculation, and by phagocytosis of these complexes and thrombocytes by various cells.