Alveolar epithelial cells in experimental lung emphysema. Ultrastructural analysis of cells in situ in TEM.

A study was made of ultrastructural changes in alveolar epithelial cells after a single intratracheal papain injection. The experiment was carried out on male Wistar rats, of 180-220 g body weight. The animals were sacrificed after 1, 7, 14 and 28 days that followed proteolytic enzyme administration. The study revealed that the development of emphysematous changes in the rat lungs was accompanied by alterations in the epithelium of alveoli. In early stages of emphysema (up to 1 week), destructing changes dominated within the epithelial cells. Similar changes were observed in the endothelium of adjacent blood vessels. In later periods an increased number of type II pneumocytes were seen and focal accumulation of elastic and collagenic fibres was noticed in alveolar septa. Correlation was found between changes in the number of type II cells and their ultrastructure as well as between alveolar epithelium damage and alveolar septal interstitium alterations.

Morphometric and ultrastructural studies of the sciatic nerve regeneration in rats intoxicated with ethanol.

The aim of the study was to examine the process of sciatic nerve regeneration and changes in the dorsal root ganglia (from which sensory fibres of the sciatic nerve extend) in animals intoxicated with ethanol. The experiment used 20 rats, divided into two groups: control and treated. The treated animals were intragastrically given 2g/kg b.w. of ethanol in 25% aqueous solution. In both groups the right sciatic nerve was transected and then sutured. After 5 months the animals were anaesthetized. The left and the right spinal dorsal ganglia-L5 and sections from the non-operated and operated sciatic nerves were collected for analysis. Ultrastructural examinations and morphometric measurements were conducted. It was found that ethanol administrated to rats inhibited regeneration of the transected and then sutured sciatic nerve, impairing the growth of axons in the transected nerve and destroying the regenerating sensory ganglion cells. The mechanism of the changes described may be associated with axonal transport disorders or with the suppressed production of biologically active substances, which affect nerve regeneration.

Exocrine cell mitochondria of the rat pancreas after lead intoxication.

The aim of the study was to evaluate alterations in exocrine cell mitochondria of the rat pancreas after lead acetate intoxication. The experiment used 45 rats divided into 2 experimental groups receiving lead acetate to drink, of lead concentration 50 and 500 mg/dm3 (ppm), and a control group given tap water. The animals from the experimental group were decapitated after 2, 4, 6 and 8 weeks, 5 rats from the control group after 8 weeks of the experiment. Rats from experimental groups decapitated after 8 weeks had lead administration stopped after six weeks and then, for two weeks tap water was given. Pancreatic sections were examined with biochemical methods for the activity of cytochrome oxidase and succinic dehydrogenase. Ultrastructural and morphometric examinations were also performed. It was demonstrated that: a) exocrine cell mitochondria are particularly predisposed to lead effect, b) intoxication of rats with lower lead doses (50 ppm) causes reversible adaptative or compensatory changes in these organelles, c) intoxication of rats with higher lead doses (500 ppm) induces irreversible ultrastructural alterations in numerous mitochondria, including damage to inner and to outer mitochondrial membranes, d) structural changes in the mitochondria in the course of lead intoxication are the morphological expression of the impairment of metabolic processes, associated with the inhibited activity of the respiratory enzymes: succinic dehydrogenase and cytochrome oxidase.

Changes of myocardial capillary density in progression of experimental lung emphysema.

A morphometrical analysis of changes in the heart of rats with experimental lung emphysema due to single intratracheal administration of papain solution has been done. Special attention has been paid to quantitative changes of capillaries and to the cardiomyocyte diameter in respective regions of the myocardium after one, three and six months following the administration of papain. A histological and morphometrical analysis of changes in the lungs has also been carried out and partial pressures of oxygen and carbon dioxide in the arterial blood of the examined animals have been marked. It has been found that a single administration of papain caused emphysematous changes in the lungs, growing more intense until the third month of the experiment. The progression of these changes corresponded to a capillary density increase in the right ventricle, interventricular septum and in the subendocardial layer of the left ventricle. The increase in the number of capillaries was interpreted as the first adaptive stage of cor pulmonale development and explained by the occurrence of angiogenesis. The changes co-existed with a decrease in partial pressure of oxygen in the arterial blood. After six months of the experiment, a reduction in capillary density in the heart regions mentioned above and a simultaneous increase in the cardiomycyte transversal diameter were observed, being the exponents of evident myocardial hypertrophy. Development of ultrastructural changes in the hearts of rats has been analysed too. Simultaneous development of changes in the right ventricle muscle and in the subendocardial layer of the heart left ventricle myocardium was observed. The role of hypoxia as a one of essential factors responsible for the observed morphologic changes has been discussed.

[Mast cell of upper gastrointestinal tract mucosa and Helicobacter pylori infection in children]. / Komórka tuczna blony sluzowej górnego odcinka przewodu pokarmowego a zakazenie Helicobacter pylori u dzieci.

Using histological, immunohistochemical and electron microscopic examinations of gastric and duodenal mucosa the authors showed the increase in the number of mast cells with ultrastructural changes characteristic of their degranulation in H. pylori infected children. The damage of various intensification in glandular and covering epithelial cells and the inflammatory response were also proved. The results suggest the role of mastocytes in the inflammation of gastric and duodenal mucosa in H. pylori infected children.

[Pneumonia in children and adolescents up to 18 years of age--a comparative analysis of the years 1976-1985 and 1986-1990]. / Zapalenie pluc u dzieci i mlodziezy do 18 roku zycia--analiza porównawcza lat 1976-1985 i 1986-1990.

An analysis was done of 1084 autopsies of children and adolescents up to 18 years of age carried out in the years 1986-1990. In the analysed period 631 cases were found of pneumonia (P) (58.2% autopsies) which was an increase of about 14% in relation to the years 1976-1985. Pneumonia was most frequently observed in the age range 1-12 months. It was the direct cause of death (primary pneumonia--PP) in 130 cases (11.9% autopsies) while in 501 cases (46.2%) it accompanied other primary diseases (secondary pneumonia--SP). In relation to the years 1976-1985 a decrease was observed of the per cent of autopsies with the diagnosis of PP (from 19.5% to 11.9%), as well as a significant increase of the per cent of SP) from 25.1% to 46.2%).

Childhood-onset dense deposit disease: a rare cause of proteinuria.

INTRODUCTION: Dense deposit disease (DDD) is a rare renal disease related to the dysregulation of the alternative pathway of the complement cascade, caused by several factors including the presence of an autoantibody to C3 nephritic factor, mutations in factor H and autoantibodies to this protein. DDD is characterized by C3 accumulation with absent or scanty immunoglobulin deposition. CASE PRESENTATION: Herein we report the case of a child with benign course of DDD, who presented with moderate proteinuria and lack of clinical symptoms without immunosuppressive treatment. Laboratory testing revealed moderate proteinuria, normal serum creatinine, total protein, and albumin levels, but significantly decreased serum C3 level. The results of renal biopsy were consistent with DDD. Genetic analysis revealed that the patient carried one copy of the H402 risk allele of factor H. The level of proteinuria did not change during the follow-up period and no nephrotic syndrome signs occurred. Renal function was stable. CONCLUSION: In conclusion, a program of urine screening for asymptomatic proteinuria and hematuria to detect children with kidney disease before they experience loss of kidney functions should be considered. Children diagnosed with DDD should have the opportunity to get treatment early on and to be followed very closely.

Expression of macrophage markers in cryoglobulinemic glomerulonephritis - a possible role of CXCL9.

PURPOSE: Cryoglobulinemic glomerulonephritis (CGGN) is a type of membranoproliferative glomerulonephritis (MPGN) that develops in patients with systemic cryoglobulinemia. To date the exact pathogenesis of CGGN remains unclear. It has been suggested that macrophages may be significant contributors to the glomerular injury in this disease. In our study we attempt to characterize the macrophages in human CGGN using classical activation and regulatory macrophage markers. MATERIAL AND METHOD: We searched our database for renal biopsy cases of CGGN. Macrophages were detected using a monoclonal anti-CD68 antibody. Two groups of macrophage markers were used: classical activation markers, including iNOS, CXCL9 and CCL20, and regulatory markers: SPHK1 and LIGHT. The stains were performed using immunohistochemical method. RESULTS: Five patients with CGGN were identified. Four patients had systemic cryoglobulinemia and two had a serological evidence of hepatitis C virus infection. In all cases the glomeruli contained numerous macrophages. Staining for activatory macrophage markers revealed a strong nuclear staining for CXCL9 in numerous cells, including those corresponding to the macrophage location. Staining for the other activatory markers, as well as staining for regulatory markers, was not significant. CONCLUSION: In this study of human CGGN we showed a striking expression of cytokine CXCL9, a classical macrophage activation marker, by the macrophages and possibly other cell types within the glomeruli. This observation points to the possible role of classically activated macrophages in the pathogenesis of MPGN. If this observation is confirmed on a larger group of patients, the cytokine CXCL9 could become a potential therapeutic target for human CGGN.

Effects of perioperative immunostimulating nutritional therapy on the phagocytic activity of blood platelets in patients with various clinical stages of gastric cancer.

PURPOSE: The aim of this study was to evaluate the total blood platelets count, fraction of phagocytizing thrombocytes (PhT%), and phagocytic index of thrombocytes (PhIT) in gastric cancer considering the stage of the disease, and perioperative immunonutrition support. METHODS: Our study included 44 patients operated for gastric cancer divided into 2 groups depending on the clinical stage, and 40 healthy volunteers -a control group. Group I included 18 patients with stage I-III locoregional malignancies and Group II included 26 patients with stage IV peritoneal dissemination. All patients received immunonutrition during the perioperative period. The phagocytic activity of blood platelets was assessed by measuring PhT% and PhIT prior to and after nutritional therapy. RESULTS: In Group I, the pre-treatment PhT% and PhIT amounted to 1.08 and 0.99, respectively, and 1.26, and 1.1 after the therapy (p<0.01). In Group II, pre-treatment PhT% and PhIT were 1.12 and 0.97, after 1.18 and 1.06, respectively (p<0.05). In the controls, PhT% and PhIT were 2.26 and 1.83, respectively, significantly higher comparing to gastric cancer patients (p<0.01). CONCLUSION: Severe impairment of the thrombocyte phagocytic activity in gastric cancer patients has been found. Phagocytic activity of blood platelets was partially improved as a result of perioperative immunonutrition both in locoregional disease and in peritoneal dissemination.

Ethanolic extract of propolis, chrysin, CAPE inhibit human astroglia cells.

PURPOSE: The aim of the present study was to examine the effect of freeze dried ethanolic extract of propolis (EEP), chrysin and caffeic acid phenethyl ester (CAPE) dependently on their concentrations on the viability and morphology of human astroglia cells line (SVGp12). MATERIAL AND METHODS: Using gas chromatography - mass spectroscopy (GC-MS) we have established the composition of lyophilisate of EEP collected in Podlasie region (Poland). After 24 h, 48 h and 72 h of exposition to EEP or its ingredients we evaluated the survivability of human astroglia cells (SVGp12) using MTT test. Morphological analysis of human astroglia cells was defined by transmission electron microscope. RESULTS: About 70 ingredients of EEP were evaluated by GC-MS. We obtained the strong decline of viability of astroglia cells SVGp12 approximately to 16% after EEP; 33% after chrysin and 25% after CAPE application. Condensed form of mitochondria observed in transmission electron microscope may indicate activation of intrinsic pathway of apoptosis induced by EEP, chrysin and CAPE in SVGp12 cell line. CONCLUSION: This study showed that EEP, chrysin and CAPE reduced viability of human astroglia cells probably due to apoptosis process.

Ultrastructural study of the submandibular gland of the rat after 6-month exposure to cadmium and zinc in drinking water.

PURPOSE: Cadmium toxicity in the exposure of the general, professional and cigarette smoking populations has been well known. From the dental point of view, it is important to find out whether and how separate and joint exposures to cadmium and zinc affect the structure and function of the submandibular gland, which is the major saliva-releasing gland. Cadmium, a particularly active xenobiotic, damages cellular metabolism at the level of various enzymatic systems of the cell, which may disturb functioning of the salivary glands. Mutual interactions of cadmium and zinc suggest a protective role of zinc through the induction of metallothionein which inactivates cadmium effect. MATERIAL AND METHODS: The aim of the study was to assess the ultrastructural picture of chosen cell organelles of the submandibular salivary gland of the rat exposed to cadmium and zinc. The study used 90 male Wistar rats, of the initial b.w. 150-180 g. The animals were exposed to cadmium and/or zinc for 6 months. Cadmium was received in aqueous solutions of cadmium chloride with drinking water at a concentration of 5 mg Cd/dm3 or 50 mg Cd/dm3. Zinc was also given in aqueous solutions of zinc chloride ad libitum at concentrations of 30 mg Zn/dm3 and 60 mg Zn/dm3. RESULTS: The ultrastructural changes in the mucous and serous cells of the submandibular salivary gland were most pronounced at cadmium concentration of 50 mg Cd/dm3 and were mainly observed in the cell nucleus, Golgi Apparatus and secretory granules of the salivary gland cells. CONCLUSIONS: 1. Exposure to cadmium induces ultrastructural changes in the submandibular gland, which are dose and time of exposure. 2. Exposure to zinc did not affect significantly the ultrastructural picture of cells of the submandibular gland. 3. Zinc administered together with cadmium reduces the intensity of ultrastructural changes in the submandibular gland.

Ultrastructural evaluation of hepatocytes membranes and changes in cytosolic enzymes distribution in methanol intoxication.

Acute methanol intoxication causes metabolic and structural disturbances of liver cells. The aim of this paper, therefore, was to evaluate the ultrastructure of liver cells membrane and the amount of lipid peroxidation products, as well as the concentration of marker enzymes of liver damage (ALT and AST) in blood serum. The experiment was done on Wistar rats which once received intragastrically 6,0 g methanol/kg b.w. as a 50% solution. The animals were decapitated 6, 12 and 24 h and 2, 5 and 7 days after the methanol administration. The liver was evaluated under transmission electron microscope and lipid peroxidation products were determined in the liver homogenate. The serum ALT and AST activity were also assayed. The biochemical results indicate the increase in lipid peroxidation products. The consequence of this is probably the membrane liver cell damage visible in the electron microscope.

Ultrastructural evaluation of lysosomes and biochemical changes in cathepsin D distribution in hepatocytes in methanol intoxication.

Methanol oxidation is accompanied by free radicals and formaldehyde formation. It is likely to cause damage of lysosomal membranes. Lysosomal ultrastructure under transmission electron microscope and biochemical localization of cathepsin D were estimated after rats intoxication with methanol. The examination was carried out 6, 12 and 24 h and 2.5 and 7 days after intoxication. Ultrastructural examination showed that methanol causes extension of Golgi apparatus cisterns and an increase in a number of lysosomes. From 12 h to 2 days lysosomes were characterized by damage of structure of membrane enclosing lysosomes. During the first days of intoxication activity of cathepsin D decreased in lysosomes and increased in cytosol. These changes may lead to uncontrolled extralysosomal proteolysis in the liver cells and to the onset of liver tissue destruction.

Intermediate cells in the rat pancreas after chronic exposure to lead.

Ultrastructural analysis of the rat pancreas following the long-term exposure to lead revealed the presence of numerous intermediate cells in animals receiving lead acetate in doses of 500 ppm and 1000 ppm for 4 and 6 weeks. The cells included acinar-beta, beta-acinar and intermediate cells containing two kinds of endocrine granules- alpha and beta, and zymogen granules.

Study on carcinogenesis in chronic cholecystitis.

Not only bile but also chronic cholecystitis may play a role in gallbladder carcinogenesis. Numerous studies have revealed a close correlation between chronic inflammation and neoplasia. The experiments were conducted on paraffin sections, obtained from 377 surgically resected gallbladders with chronic cholecystitis. Immunohistochemical reaction was conducted on deparaffinized sections, using a monoclonal antibody against 8-hydroxydeoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage. An increase was found in the expression of 8-hydroxydeoxyguanosine in chronic cholecystitis. The level of 8-OhdG expression is associated with inflammation intensity and disease duration. DNA damage, observed in chronic cholecystitis, indicates a correlation between chronic inflammation and gallbladder carcinogenesis.

A preliminary study of the submandibular gland of the rat after one-year cadmium intoxication. Part II. Pathomorphology and ultrastructure.

The aim of the present study was to establish to what degree a one-year exposure of rat females to 5, 50 and 100 mg of Cd/l affects cell morphology of the submandibular glands. After one-year cadmium exposure of female rats, at doses of 5, 50 and 100 mg Cd/l, a pathomorphological examination revealed periductal fibrosis in the submandibular glands of rats in all the three experimental groups, which increased with cadmium dose. We also found foamish cytoplasm in the cells of the submandibular glands in all the experimental groups, the intensity of that phenomenon also increasing with Cd dose. The ultrastructural examination revealed no abnormalities in Group I. However, in Groups II and III, we observed numerous granules with a secretion, varying in shape and size that filled up the cytoplasm both in the mucous and serous cells.

Ultrastructural evaluation of the rat parotid gland after six-week-intoxication with lead acetate.

Clinical and experimental data point to the unfavorable effect of lead compounds on the oral cavity condition. The aim of this study was to analyse ultrastructural changes in the parotid gland of rats subjected to intoxication with lead acetate. The experiment was carried out on 24 Wistar rats divided into 4 groups, 6 animals in each. Group I animals were given aqueous solution of lead acetate of lead concentration 50 mg/dm3(50ppm), group II-500 mg/dm3(500ppm), group III-1,000 mg/dm3(1,000ppm). Control animals drank tap water. All the rats were sacrificed after six weeks of the experiment and material was collected for ultrastructural studies and evaluation of lead level in the tissue dry mass. A three fold increase in lead levels in the parotid gland was observed in group I and a tenfold increase in groups II and III. In all experimental ultrastructural examinations revealed groups in which lead impaired secretion of alveolar cells, damaged mitochondria and disturbed the lipid balance. The greatest destructive changes were observed in group III.

Ultrastructural evaluation of the rat parotid gland in the course of intoxication with lead acetate.

The purpose of this study was to evaluate the ultrastructural changes in the parotid gland of rats exposed to lead. The animals were given lead acetate in drinking water, at a concentration of 50 mg/dm3 Pb for periods of 2, 4 and 6 weeks. The ultrastructural examination showed that lead especially impaired mitochondria, caused accumulation of lipid droplets in the cytoplasm of acinar cells and inhibited the process of exocytosis of secretory granules. In the most extreme cases some acinar cells were destroyed.