Prognostic factors in oral and oropharyngeal cancer based on ultrastructural analysis and DNA methylation of the tumor and surgical margin.

Oral and oropharyngeal cancers are characterized by relatively low 5- year survival rates due to many factors, including local recurrence. The identification of new molecular markers may serve for the estimation of prognosis and thus augment treatment decisions and affect therapy outcome. The aim of this study was to describe the morphological characteristics and the DNA methylation status of the CDKN2A,CDH1, ATM, FHIT and RAR- genes in the central and peripheral part of the tumor and the surgical margin and evaluate their prognostic significance. 53 patients with oral and oropharyngeal cancer were enrolled to the prospective study, and had been primarily treated surgically. Correlations between morphological data, hypermethylation status and clinicopathological data, as well as prognosis, were assessed. Nuclei polymorphism highly correlated with T stage (p < 0.0001), N stage (p < 0.046), and metastases to the lymph nodes pN (p < 0.004 ). Also, the number of cells in irregular mitosis correlated with T stage (p < 0.004), and highly with pN (p < 0.009). The significance of CDKN2A hypermethylation as a good prognostic factor was also established in the Kaplan-Meir test. The ultrastructural analysis showed that none of the examined tumors had homogenous texture and that resection margin specimens clean in HE stained tissue samples frequently contained single tumor cells or few cells in groups surrounded by connective tissue. This indicates the superiority of electron microscopy over standard histopathological analysis. Thus, a combination of such morphological examination with epigenetic parameters described herein could result in the discovery of promising new prognostic markers of the disease.

Effects of L-arginine and NG-nitro L-arginine methyl ester (L-NAME) on ischemia/reperfusion injury of skeletal muscle, small and large intestines.

This study analyzed the effects of L-arginine and non-specific nitric oxide (NO) synthase blocker (L-NAME) on structural and metabolic changes in experimental ischemia/reperfusion injury in the rat. Histopathological evaluation of rat tissues after reperfusion was also performed. The animals were divided into four groups: [1] nonischemic control, [2] ischemia 4 hrs/repefusion 30, 60, 120 min, [3] ischemia/reperfusion after L-arginine administration, [4] ischemia/reperfusion, after L-arginine, and L-NAME. L-arginine (500 mg/kg) and L-NAME (75 micromol/rat/day) were administrated orally for 5 days before experiment. Concentrations of free radicals, CD-62P, CD-54 and malonyl dialdehyde (MDA) in tissues, and MDA and NO levels in sera were determined. Free radical levels significantly increased in reperfused skeletal muscle, small and large intestines. In large bowel, reperfusion increased MDA levels and evoked a rise of endotoxin level while NO levels decreased. Histological studies showed an increase in the number of lymphocytes in both intestines. Administration of L-arginine reduced leukocyte adherence associated with ischemia-repefusion injury, decreased the levels of free radicals and MDA in the examined tissues, and inhibited the release of endotoxins into blood. L-arginine-treated animals showed higher serum NO levels and reduced leukocyte bowel infiltration. Concomitant L-NAME administration reduced serum NO and tissue free radical [corrected] levels, but did not affect intestinal leukocyte infiltration. L-arginine could ameliorate intestinal ischemia/reperfusion injury and constitute a possible protective mechanism by decreasing neutrophil-endothelial interactions, stimulating free radical scavenging and reducing lipid peroxidation.

Histological alterations of gallbladder mucosa and selected clinical data in young patients with symptomatic gallstones.

Histological lesions of gallbladder were described mainly in older patients with cholelithiasis (CH). The aim of the study was to analyse morphological alterations in gallbladder mucosa and selected clinical data of young patients with CH. The studies were conducted on 57 patients with CH, subjected to cholecystectomy in the years of 2003-2007. In course of the years, 37 respective young patients (below 25 years of age) were operated. The comparative group included twenty 50-year-old patients with gallstones. The inflammatory activity (grading) was evaluated using a semiquantitative scale on HE-stained gallbladders. In either group, women with chronic cholecystitis and multiple gallstones prevailed. Histological alterations in young patients involved absence of evident epithelial metaplasia traits, low number of foamy cells and prevalence of eosinophils in gallbladder mucosa. Even if a similar grading in gallbladder walls was noted in young and older patients, only in the former ones, a higher grading was detected in patients with an acute clinical course of the gallstone disease. The results point also to a potential role of local accumulation of eosinophils in gallbladder mucosa in pathogenesis of CH in young patients.

[Changes in tracheal ciliated epithelium of rats exposed to environmental tobacco smoke--experimental studies]. / Zmiany w nablonku migawkowym tchawicy szczurów pod wplywem ekspozycji na dym tytoniowy--badania doswiadczalne.

It has been already proved in many experimental studies that tobacco smoke has multiple toxic effects on respiratory tract cells. Alterations in ciliary epithelium of rats trachea after short exposition to high tobacco smoke concentrations in inhaled air were been determinate in current study. Morphological evaluation revealed in lining epithelium voluminous exudate located between epithelium and lamina propria cells with evidently larger number of mast cells.

Carvedilol Inhibits Matrix Metalloproteinase-2 Activation in Experimental Autoimmune Myocarditis: Possibilities of Cardioprotective Application.

AIMS: Acute myocarditis is a potentially lethal inflammatory heart disease that frequently precedes the development of dilated cardiomyopathy and subsequent heart failure. At present, there is no effective standardized therapy for acute myocarditis, besides the optimal care of heart failure and arrhythmias in accordance with evidence-based guidelines and specific etiology-driven therapy for infectious myocarditis. Carvedilol has been shown to be cardioprotective by reducing cardiac pro-inflammatory cytokines present in oxidative stress in certain heart diseases. However, effects of carvedilol administration in acute myocarditis with its impact on matrix metalloproteinases' (MMPs) activation have not been elucidated. METHODS AND RESULTS: Carvedilol in 3 doses (2, 10, and 30 mg/kg) was given daily to 3 study groups of rats (n = 8) with experimental autoimmune myocarditis by gastric gavage for 3 weeks. In comparison to untreated rats (n = 8) with induced myocarditis, carvedilol significantly prevented the left ventricle enlargement and/or systolic dysfunction depending on the dose in study groups. Performed zymography showed enhanced MMP-2 activity in untreated rats, while carvedilol administration reduced alterations. This was accompanied by prevention of troponin I release and myofilaments degradation in cardiac muscle tissue. Additionally, severe inflammatory cell infiltration was detected in the nontreated group. Carvedilol in all doses tested, had no impact on severity of inflammation. The severity of inflammation did not differ between study groups and in relation to the untreated group. CONCLUSIONS: The protective effects of carvedilol on heart function observed in the acute phase of experimental autoimmune myocarditis seem to be associated with its ability to decrease MMP-2 activity and subsequently prevent degradation of myofilaments and release of troponin I while not related to suppression of inflammation.

Selected markers (chromogranin A, neuron-specific enolase, synaptophysin, protein gene product 9.5) in diagnosis and prognosis of neuroendocrine pulmonary tumours.

Neuroendocrine tumours of lungs represent a subgroup of pulmonary tumours with typical morphofunctional traits. In light microscopy, the four principal types of the tumours (typical and atypical carcinoids, small cell lung cancer, large cell neuroendocrine carcinoma) demonstrate typical arrangement of cells (organoid nesting, palisading, a trabecular pattern, and rosette-like structures), variable number of mitoses, presence or absence of necrosis. In ultrastructure, neuroendocrine tumours manifest groups of cells with cytoplasmic granules (and the so called dense-core neurosecretory granules in particular). Neuroendocrine cells release hormones to circulation or in a paracrine manner. Some pulmonary tumours exhibit no neuroendocrine morphology at the level of light microscopy but demonstrate ultrastructural and/or immunohistochemical traits of neuroendocrine differentiation. Proteins the presence of which confirms neuroendocrine origin of the tumours have been found relatively early to include neuron-specific enolase (NSE), the group of chromogranins and synaptophysin. Present study aimed at summing up results of investigations conducted in, approximately, recent 30 years pertaining expression and/or serum concentrations of four neuroendocrine markers (chromogranin A, neuron-specific enolase, synaptophysin, protein gene product 9.5) and at an attempt to evaluate the role of such studies in extension of diagnostic and prognostic potential as related to neuroendocrine pulmonary tumours. Until now, the most sensitive and specific marker or marker combination for early detection of neuroendocrine subtypes of lung tumours has not been identified. All of the markers examined in present study were detected both in the typical neuroendocrine pulmonary tumours and in a certain proportion of non-endocrine tumours. In the case of chromogranin A improved sensitivity and specificity of immunocytochemical studies was obtained using a panel of antibodies directed to various epitopes of the protein. Both in endocrine and non-endocrine tumours, neuron-specific enolase (NSE) is thought to represent mainly a prognostic index, and only quantitation of serum concentrations of the protein or of the fraction of immunopositive cells may permit to differentiate between subtypes of the tumours. Synaptophysin is regarded to represent one of the most specific markers of neuroendocrine differentiation, manifesting a much higher sensitivity than chromogranin A and NSE. With increasing frequency, PGP 9.5 is regarded to provide a prognostic marker in diagnosis of non-small cell lung carcinomas rather than of typical neuroendocrine tumours.

[Antiplatelet therapy can unmask an inherited bleeding disorder. Aspirin-like defect of platelets does not protect against atherosclerosis]. / Leczenie przeciwplytkowe moze ujawnic wrodzona skaze krwotoczna. Aspirynopodobny defekt plytek krwi nie chroni przed miazdzyca

Some inherited platelet disorders may be revealed late, as in the presented case of a 68-year-old-man. Recurrent epistaxis following peri-interventional antiplatelet therapy (after three elective percutaneous coronary interventions) and an episode of upper gastrointestinal haemorrhage required aspirin withdrawal and less frequent clopidogrel use. Platelet studies showed an aspirin-like defect resulting in a lack of arachidonate-induced platelet aggregation. During dose-reduced (2-3 times a week) clopidogrel administration ADP-induced platelet aggregation was effectively inhibited and neither important bleeding nor stent thrombosis occurred. The inherited defect of cyclooxygenase-1, responsible for platelet thromboxane synthesis, did not protect the patient against coronary and extra-cardiac atherosclerosis.

[The role of immunohistochemical staining (protein p53, cyclin D1) in the prognosis of adenoid cystic carcinoma salivary gland tumors]. / Rola badan immunohistochemicznych (bialka p53, cykliny D1) w prognozowaniu raka gruczolowato-torbielowatego (carcinoma adenoides cysticum) slinianek.

INTRODUCTION: Adenoid cystic carcinoma of the salivary glands (carcinoma adenoides cysticum) is malignant epithelial tumor of rare occurrence. Tumor of this kind has among salivary glands tumors uncertain prognosis and unpredictable course. The aim of the study was to characterize the patient population and the immunohistochemical analyses (p53 protein, cyclin D1). MATERIAL AND METHODS: The examined group consisted of 30 patients with adenoid cystic carcinoma of the salivary glands. The expression of p53 protein and D1 cyclin in the tumor was evaluated and the correlation between these proteins and the organ and clinical grading was defined. RESULTS: The immunohistochemical studies showed in 70% the positive staining for p53 protein and 90% for cyclin D1. There was not statistically significant difference between the advanced grading of the organic and clinical adenoid cystic carcinoma.

Analysis of the impact of harmful factors in the workplace on functioning of the respiratory system of firefighters.

INTRODUCTION AND OBJECTIVE: Firefighters are considered a healthy and fit group of individuals, well-prepared for taking action in disaster situations. While working, they suffer from exposure to certain toxic agents, especially combustion products generated when a fire takes place. Among them, the most frequent and the most toxic are: carbon monoxide, hydrogen cyanide, ammonia, and those resulting from PVC combustion - hydrochloride, phosgene and chloride. Additionally, fire-extinguisher powder can be inhaled and lead to certain lesion in the airways. The aim of study was to ascertain the influence of toxic agents present at the scene of fire on the lung tissue of firefighters, and also to study this on an animal model. MATERIAL AND METHODS: The study group consisted of firefighters who had a minimum of 10 years service. After completing a questionnaire, their clinical status was ascertained based on a general examination, laboratory tests and lung function tests. RESULTS: Questionnaire analysis showed a high percentage of pathological symptoms in the studied group. The incidence of the symptoms correlated with the duration of occupational exposure to toxic agents. Among other results, obstruction of flow in medium airways in about 30% of the studied individuals represented the most important finding. Experimental tests were next performed on male Wistar rats, aged 3 months. They were insufflated with the solution of powdered fire-extinguisher, after which morphology specimens of lung tissue were studied. Evidence for disseminated fibrosis was obtained, which supported the previous clinical findings in the firefighters. CONCLUSIONS: The above shows correlation between occupational exposure and respiratory system involvement in firefighters. This justifies covering the group of firefighters with special medical care focused on prophilaxis, early detection and therapy of pulmonary diseases.

Does loss of heterozygosity in critical genome regions predict a local relapse in patients after laryngectomy?

BACKGROUND: Patients, who had an upper aerodigestive tract malignancy, have a high incidence of succeeding tumor development. This has been attributed to the role of "field cancerization" in carcinogenesis. The aim of this study was analysis of loss of heterozygosity (LOH) in the regions frequently lost during the course of head and neck squamous cell carcinomas (HNSCC), especially at early stages, which could answer the clinicians' question, if LOH analysis has any "predictive" value in relation to tumor occurrence. MATERIAL AND METHODS: Sixty-five larynx cancer patients were examined for loss of heterozygosity on 3p, 7q, 8p, 9p and 18q chromosomal arms with the use of 12 microsatellite markers. The material from a single patient consisted of blood, tumor, safe margin and one or two clinically unchanged mucosal samples. During follow up, the material from brush specimens (14 patients) as well as laryngeal swabs (4 patients) was also examined. RESULTS: The highest frequency of LOH was detected for marker D3S1234 in tumor tissues (29%). Analysis of margin samples (b) revealed low LOH frequencies (2-5%) and complete retention of heterozygosity for markers: D3S1234, D7S486, D8S261, D8S264, D9S171 and D18S46. Similarly, for normal appearing mucosa from upper part of larynx (c) frequencies of LOH were low (2-6%), with the complete retention of heterozygosity for markers: D3S1284, D3S1304, D3S1234, D8S264 and D9S1870. We did not detect any LOH in the material of normal appearing mucosa from tracheostoma region (d). During follow up, LOH was detected for eight markers, with the highest incidence for markers D18S46 (six cases), D7S486 (four cases) and D3S1300 (three cases). CONCLUSIONS: The data, obtained during this investigation, did not reveal the predictive value of LOH with respect to local relapse occurrence in laryngeal cancer patients. However, time of follow up did not reach 5 years, so that further clinical monitoring should be conducted.

Interleukin-2 (IL-2) expression in livers of patients with chronic hepatitis C virus (HCV) infection.

The studies performed till now have pointed to an increased serum levels of interleukin 2 (IL-2) in infection with hepatitis C virus (HCV). The present study was aimed at examining intrahepatic expression of IL-2 in children (n=15) and in adults (n=11) with chronic hepatitis C as well as its correlations with histological lesions and selected clinical data. The immunocytochemical techniques and in situ hybridization method were applied at light and electron microscopy level. Under the light microscope, expression of IL-2 was analysed semiquantitatively. As compared to the control material, in livers of both groups of chronic hepatitis C patients augmented expression of IL-2 was demonstrated. The reaction product was localized mainly in the cytoplasm of hepatocytes which was confirmed by hybridocytochemistry. The mean proportion of cells with positive reaction for IL-2 mRNA was significantly lower than the proportion of cells positive for the respective protein. No correlation was disclosed between IL-2 expression on one hand and grading or staging, alanine aminotransferase (ALT) and HCV RNA levels in serum on the other. At the ultrastructural level, IL-2 in hepatocytes was present mainly in the endoplasmic reticulum and mitochondria. Our studies have confirmed augmented expression of IL-2 in livers of patients with chronic hepatitis C and have demonstrated that hepatocytes represent the principal source of the cytokine in HCV in vivo infection. Moreover, expression of IL-2 in the infection was examined for the first time at the ultrastructural level. Mitochondrial localization of IL-2 suggests a direct involvement of the cytokine in disturbed function of the organelles.

Long-term consequences of total gastrectomy: quality of life, nutritional status, bacterial overgrowth and adaptive changes in esophagojejunostomic mucosa.

OBJECTIVES: The aim of the study was to evaluate long-term quality of life and adaptive changes in the mucosa of the proximal section of the small intestine used for esophagojejunostomy reconstruction in stomach cancer patients after total gastrectomy. MATERIAL AND METHODS: Thirty-one patients who had undergone stomach cancer-related total gastrectomy were included in the study, which spanned a period of 48 to 127 months (79.6 months on the average) after the surgery. The analysis included: a) evaluation of selected biochemical parameters; b) microbiological evaluation of esophagojejunostomic area; c) evaluation of adaptive changes in esophagojejunostomic mucosa using light and electron microscopy; d) quality of life evaluation with a Troidl questionnaire. RESULTS: Quality of life was subjectively rated as good or very good by almost all subjects. The analyzed biochemical parameters were within the range of normal values in all the subjects with the exception of mild abnormalities in alkaline phosphatase and vitamin B12 levels in some patients. Microbiological examination of mucosal specimens from below the esophagojejunostomy revealed significant bacterial flora overgrowth in all the patients, with streptococci being the most abundant species. Light and electron microscopy examination of the epithelium confirmed it was normal and characteristic of a healthy small intestine. CONCLUSIONS: Long-term quality of life in patients after complete stomach resection is considered good or very good, irrespective of the reconstruction method used, and the esophagojejunostomic mucosa of the reconstructed area is normal and typical for a healthy small intestine.

Morphological and molecular diagnosis of a fatal form of EBV infectious mononucleosis in a child.

Mortal cases of acute Epstein-Barr virus (EBV) infection in the form of mononucleosis have been seldom described and used to be related to complications of the disease. In this report, the case of a 3-year-old girl is described, with severe form of infectious mononucleosis, deceased in the course of respiratory-circulatory insufficiency with a sudden cessation of heart action. Particular attention was given to histological lesions, phenotype of inflammatory cells and to expression of proteins and EBV RNAs (EBERs) in tissues, examined using immunocytochemical techniques and in situ hybridization. Histological patterns were dominated by massive lymphocyte infiltrates (mostly CD45RO+ and CD3+ cells), mainly in lungs and in liver and, less pronounced, in kidneys and in leptomeninx. Lymphocyte proliferation exhibited polyclonal character: both lambda and kappa chains were present. No myeloblastic differentiation could be demonstrated. The EBV proteins, as well as EBV RNAs (EBERs) were detected both in small lymphocytes B and in enlarged (blast) cells, frequently resembling Reed-Sternberg cells. In our tissue material co-expression of the two proteins (EBNA2+, LMP1+) and EBER has been demonstrated in every organ, in accordance to the latency III pattern described by other authors.

Expression of cytokines (TNF-alpha, IL-1alpha, and IL-2) in chronic hepatitis C: comparative hybridocytochemical and immunocytochemical study in children and adult patients.

Hepatitis C virus (HCV) is one of the principal causes of hepatitis, which in more than 80% of cases leads to chronic lesions in the liver and involvement of extrahepatic organs. It remains unknown why the infection so frequently turns chronic, independently of patient age. Using immunocytochemistry (IHC) and in situ hybridization (ISH) (both linked to the ImmunoMax technique) we examined cell sources of TNF-alpha, IL-1alpha, and IL-2 in control and HCV-infected children and adults. We demonstrated augmented expression of all the cytokines in HCV-infected patients compared to controls. No differences were detected in amounts of studied transcripts or cytokine proteins between biopsies taken from HCV-infected children and adults. Expression of TNF-alpha was localized mainly in liver sinusoidal cells (macrophages, endothelial cells). A high proportion of hepatocytes demonstrated expression of TNF-alpha, IL-1alpha, and IL-2. In both groups of patients, higher amounts of cytokine proteins than studied transcripts were demonstrated. The augmented expression of TNF-alpha, IL-1alpha, and IL-2 in liver with a similar proportion of involved cells (mainly hepatocytes) in children and in adults points to participation of the cytokines in the pathogenesis of chronic hepatitis C. The expression is insufficient to terminate the infection and may be linked with the comparably frequent chronic transformation of HCV infection noted in children and adults.

Oxidative DNA base modifications and polycyclic aromatic hydrocarbon DNA adducts in squamous cell carcinoma of larynx.

Tobacco smoke, recognized as a major etiological factor for cancers of the upper aerodigestive tract, represents an abundant source of reactive oxygen species (ROS), which are believed to play a significant role in mutagenesis and carcinogenesis. An additional source of ROS in tissues exposed to tobacco smoke may be metabolic oxidation of polycyclic aromatic hydrocarbons (PAH). To investigate the relationships between oxidative DNA lesions and aromatic DNA adducts, six modified DNA bases 5-hydroxyuracil, 5-hydroxycytosine, 7,8-dihydro-8-oxoguanine, 7,8-dihydro-8-oxoadenine, 2,6-diamino-4-hydroxy-5-formamidopyrimidine and 4,6-diamino-5-formamidopyrimidine and the total level of PAH-related DNA adducts were measured in cancerous and the surrounding normal larynx tissues (68 subjects), using gas chromatography/isotope-dilution mass spectroscopy with selected ion monitoring and the 32P-postlabeling-HPLC assay, respectively. The levels of oxidative DNA lesions in cancerous and adjacent tissue were comparable; the differences between the two types of tissue were significant only for 5-hydroxypyrimidines (slightly higher levels were observed in the adjacent tissue). Comparable levels of DNA lesions in cancerous and the surrounding normal tissues observed in the larynx tumors support a field cancerization theory. The surrounding tissues may still be recognized as normal by histological criteria. However, molecular alterations resulting from the chronic tobacco smoke exposure, which equally affects larynx epithelia, may lead to multiple premalignant lesions. Thus, a demonstration of similar levels of DNA damage in cancerous and the adjacent tissue could explain a frequent formation of secondary tumors in the larynx and the frequent recurrence in this type of cancer. A weak, but distinct effect of tumor grading and metastatic status was observed in both kinds of tissue in the case of 5-hydroxyuracil, 5-hydroxycytosine, 7,8-dihydro-8-oxoguanine, 7,8-dihydro-8-oxoadenine. This effect was displayed as a gradual shift in the data distribution toward high values from G1 through G2-G3 and from non-metastatic to metastatic tumors. Since the levels of oxidative DNA base modifications tended to increase with the tumor aggressiveness, we postulate that the oxidative DNA lesions increase genetic instability and thus contribute to tumor progression in laryngeal cancer. No associations between aromatic adduct levels and oxidative DNA lesions were present, suggesting that the metabolism of PAH does not contribute significantly to the oxidative stress in larynx tissues, remaining the tobacco smoke ROS as a major source of oxidative DNA damage in the exposed tissue.

Effect of exogenous pulmonary surfactant preparations on the structure of pulmonary alveoli in newborn rats.

Treatment of pre-term newborns with exogenous surfactant preparation is a well established part of the therapy for respiratory distress syndrome of the newborns (RDS). Since the introduction of surfactant into clinical practice in 1980, hundreds of studies have been published describing beneficial effects of such treatment. There is only limited number of morphological publications reporting adverse effects of surfactant administration. The aim of the present study is to describe morphological changes in the lung after surfactant administration to healthy newborn rats. Two types of surfactant were used: Exosurf (Glaxo Wellcome, England) and Survanta (Abbott Laboratories, USA). Surfactant preparation were given intratracheally in single dose (bolus) (100 mg of lipids per kg b.w.). Animals from control group received 0.9% saline in equivalent volume. Lung specimens were taken 15, 20, 25 and 30 minutes after drug administration and evaluated by light and electron microscopy. There was no damage in lungs from the control group. Tissue specimens from the Exosurf group revealed severe pathological changes: foci of atelectasis, frank edema in the parenchyma, focal disruption of air-blood barrier, hemorrhages in many alveoli, surfactant particles in many alveolar capillaries, and strongly activated alveolar macrophages. In this group changes appeared as early as 15 min after surfactant administration and intensity of lung injury increased with time. Also, Survanta administration caused damage to the lung tissue. However, the changes were less intense and appeared later (20-25 minutes after Survanta treatment). In conclusion, the presented morphological findings proved that exogenous surfactant administration to healthy rat newborns caused lung damage. Comparing two different surfactant preparation, Exosurf and Survanta, it was shown that the former one produced stronger and faster damage to lung alveoli than the latter one.

Morphological lesions detected by light and electron microscopies in chronic type B hepatitis.

Apart from serological diagnosis of chronic type B hepatitis, of high importance is specific morphological diagnosis, based on evaluation of liver biopsies. In the evaluation, the techniques are employed, which directly visualize the virus in the cells (electron microscopy) as well as the techniques of cell biology which demonstrate the presence of viral genetic material and viral proteins in situ. This paper reviews the available data on the diagnosis of liver pathomorphology using the above mentioned techniques in chronic HBV infections in adults. The data have been compared with the results of our own studies, performed in children. In chronic type B hepatitis and more frequently in children than in adults, slight or moderate inflammation (grade 1 to 2) and insignificantly advanced fibrosis (stage 1 to 2) are noted in the liver. Both in children and in adults, lesions in hepatocyte nuclei represent the common morphological denominator in the patterns of light and electron microscopy. The cell nuclei are of variable size, irregular shape, they stain irregularly, manifest an altered outline of nuclear envelope, frequently exhibit numerous and enlarged cell nucleoli and chromatin dissociation (the so called empty cell nuclei). In ultrastructural studies, hepatocyte cytoplasm contains Dane's bodies and tubular forms of HBsAg while virus-resembling particles are noted in cell nuclei. Molecular biology techniques (immunocytochemistry, in situ hybridisation) reveals nuclear and/or cytoplasmic location of HBcAg, cytoplasmic location of HBsAg and a similar location of HBV DNA. The data permit us to determine precisely the stage of infection and to make appropriate therapeutic decisions.

Morphological lesions detected by light and electron microscopies in chronic type C hepatitis.

HCV infection results in chronic hepatitis in most patients. The mechanisms determining liver damage and the events that lead to a high rate of chronic hepatitis remain unclear. In present study, an attempt was made to sum up data on lesions in the liver in the course of chronic type C hepatitis including those of our own cases, because that pattern is still a matter of debate. Cell lesions detected by light microscopy are characteristic but not specific and included inflammatory lesions of low or moderate intensity and a mild extent of fibrosis in the liver. The common and most characteristic trait of chronic HCV infection involves lesions in hepatocyte nuclei. These changes involved swelling, altered shape, hyperchromasia, disturbed nuclear chromatin structure, enlarged and frequently multiple nucleoli and lesions of nuclear envelope. Complexes of tubules or branching fibrils of 20-30 nm in diameter were present in cell nuclei at electron microscope level. The nuclear lesions were accompanied in the same cells by changes in rough endoplasmic reticulum with long tubular structures or branching fibrills inside. Other cytoplasmic changes included mitochondrial lesions, numerous lipid vacuoles and free tubular structure of a highly osmophilic character. Cellular localisation of HCV proteins using immunocytochemical techniques remains to be a matter of studies. In most studies HCV proteins have been detected in the cytoplasm although some reports indicate nuclear localisation, especially of C protein. All our observations on morphological lesions in chronic type C hepatitis can generally confirm most of data of other authors, but the criteria of nuclear lesions defined at the ultrastructural level represent the original input of our studies. The studies using molecular biology techniques should be continued at the electron microscope level.

Localisation of exogenous surfactants in cell membranes in the air-blood barrier: rat model.

The use of exogenous surfactants has been introduced into the therapy of patients of different ages. Much better results have been obtained in the treatment of respiratory distress syndrome with surfactants enriched with surfactant proteins. In the following study we used protein-containing surfactants (survanta and curosurf). The aim of the following study was to determine the localisation of artificial surfactants in the lung tissue. Using the Immunogold Technique, biotinylated surfactant proteins were traced in the air-blood barriers. In all lungs the exogenous surfactant was present only in some alveoli. In these parts small areas of atelectasis as well as oedema and transudate accumulation were seen. These changes were less severe after biotinylated curosurf treatment. In electron microscope studies we found surfactant elements in the air-blood barrier and other structures of the alveolar septa. Immunogold studies confirm the presence of biotynylated surfactant in the elements of the air-blood barrier.

Changes in the bronchial epithelia in patients with immotile cilia syndromes.

Immotile cilia syndromes is a cause of recurrent infection of the airways and recurrent bronchopneumonias. Among the ciliary abnormalities are found changes in the structure of the microtubules, unco-ordinated ciliary movements caused by the absence of inner or outer or both dynein arms, and abnormalities of the kinetosomes and/or rete ridges. In patients with ciliary dyskinaesia bronchitis occurs early in life (during infancy) and usually has a recurrent tendency, so that bronchial biopsy is frequently undergone for diagnostic purposes. In this study we include 127 bronchial biopsies from patients (from 2 months to 49 years) unsuccessfully treated for recurrent respiratory tract infections. When performing regular diagnostic procedures on the light and electron microscopic level, we have looked for cilia abnormalities and also focused on changes within the mucosa and submucosa. The most common abnormality recorded was absence of the inner dynein arms, but in 40 cases neither of the dynein arms were present. Only a few patients had classical Kartagener's syndrome. Special attention is drawn to biopsies from elderly patients, in whom long-standing infections were followed by extensive damage to the bronchial epithelium, including even a total absence of ciliated cells. In some cases enhanced regenerative processes and some foci of squamous metaplasia were found. In two cases even foci of low-grade dysplasia were diagnosed.