p21/Wafl/Cipl cellular expression in chronic long-lasting hepatitis C: correlation with HCV proteins (C, NS3, NS5A), other cell-cycle related proteins and selected clinical data.

Studies indicate that proteins of hepatitis C virus (HCV) disturb expression of cell-cycle-related proteins. A disturbed cell-cycle control is a hepatocellular carcinoma (HCC) risk factor in patients with HCV-related liver damage. The present study aimed to analyse the cellular expression of p21/Wafl/Cipl (p21) in long-lasting chronic hepatitis C (CH-C), its correlation with the key oncogenic HCV proteins (C, NS3, NS5A), other cell-cycle-related proteins (PCNA, Ki-67, cyclin D1, p53) and selected clinical data. Archival liver biopsies, obtained from patients with CH-C, normal livers, and hepatocellular carcinoma (HCC) specimens were analysed by immunocytochemistry and ImmunoMax technique. In CH-C overexpression of p21 protein was demonstrated. Positive correlations of p21 protein expression in CH-C involved age of the patients, grading, and liver steatosis. Moreover, expression of p21 correlated significantly with expression of p53 protein, of D1 cyclin and Ki-67. Although Ki-67 antigen was related to p21 expression, only Ki-67 expression proved to be directly related to liver staging. Expression of the NS3 protein, which prevailed in CH-C patients, manifested correlation with p21 expression, and that of cyclin D1. In presence of preserved potential for regeneration, overexpression of p21 indicates inhibition of cell cycle in hepatocytes, which probably plays a protective role for the chronically damaged cells. Out of the three HCV proteins only NS3 seems to affect control of p21 protein expression in in vivo infection. Nevertheless, the studies indicate that neither expression of p21 protein nor that of viral NS3 protein can serve as a marker of progression of CH-C to HCC in vivo

p53 immunocytochemistry and TP53 gene mutations in patients with chronic hepatitis C virus (HCV) infection.

Chronic infection with hepatitis C virus (HCV) is regarded as a risk factor for hepatocellular carcinoma (HCC), mostly in patients with liver cirrhosis. Present study aimed at evaluation of cellular expression of p53 protein, genetic TP53 changes in liver samples and anti-p53 in serum of patients with chronic hepatitis C virus infection. The expression of p53 protein were analysed by immunocytochemistry in liver biopsies from adult patients with chronic, long-lasting hepatitis C. In order to detect TP53 mutations, PCR/SSCP and sequencing were performed. Antibodies against p53 in serum were determined using enzyme immunoassay (ELISA).In two out of 14 examined patients TP53 point mutations were detected in the liver samples. In the first patient, a substitution of C to T was demonstrated in position 1 of the codon 250, resulting in substitution of proline by serine. The other patient carried a substitution of C to G in position 13274 of the intron 6. The patient carrying mutation in the codon 250 demonstrated morphological traits of liver cirrhosis and had high number of p53-immunoreactive cell nuclei in tissue. None of the patients manifested elevated titres of serum anti-p53. In the liver, significant positive correlations were disclosed between expression of p53 on one hand and grading and staging on the other. A negative correlation was disclosed between cellular expression of p53 and duration time of infection. In conclusions, genetic changes in TP53 can be detected also in non-neoplastic lesions linked to chronic HCV infection

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

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

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 -30nm 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

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

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

p21/Wafl/Cipl cellular expression in chronic long-lasting hepatitis C: correlation with HCV proteins (C, NS3, NS5A), other cell-cycle related proteins and selected clinical data.

Studies indicate that proteins of hepatitis C virus (HCV) disturb expression of cell-cycle-related proteins. A disturbed cell-cycle control is a hepatocellular carcinoma (HCC) risk factor in patients with HCV-related liver damage. The present study aimed to analyse the cellular expression of p21/Wafl/Cipl (p21) in long-lasting chronic hepatitis C (CH-C), its correlation with the key oncogenic HCV proteins (C, NS3, NS5A), other cell-cycle-related proteins (PCNA, Ki-67, cyclin D1, p53) and selected clinical data. Archival liver biopsies, obtained from patients with CH-C, normal livers, and hepatocellular carcinoma (HCC) specimens were analysed by immunocytochemistry and ImmunoMax technique. In CH-C overexpression of p21 protein was demonstrated. Positive correlations of p21 protein expression in CH-C involved age of the patients, grading, and liver steatosis. Moreover, expression of p21 correlated significantly with expression of p53 protein, of D1 cyclin and Ki-67. Although Ki-67 antigen was related to p21 expression, only Ki-67 expression proved to be directly related to liver staging. Expression of the NS3 protein, which prevailed in CH-C patients, manifested correlation with p21 expression, and that of cyclin D1. In presence of preserved potential for regeneration, overexpression of p21 indicates inhibition of cell cycle in hepatocytes, which probably plays a protective role for the chronically damaged cells. Out of the three HCV proteins only NS3 seems to affect control of p21 protein expression in in vivo infection. Nevertheless, the studies indicate that neither expression of p21 protein nor that of viral NS3 protein can serve as a marker of progression of CH-C to HCC in vivo

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