[Intranuclear particles in chronic non-A, non-B hepatitis: what diagnostic significance?]. / Particelle intranucleari in corso di epatite cronica NANB: quale utilità diagnostica?

Intranuclear particles of 23-27 nm diameter have been repeatedly demonstrated in the nuclei of hepatocytes of patients with non-A, non-B hepatitis and of experimentally infected chimpanzees; however, their specificity has been challenged since they have also been observed in other pathological conditions and in healthy volunteers. We have conducted an ultrastructural study of liver biopsies from 10 patients with chronic active non-A, non-B hepatitis. The intranuclear particles, which were observed in all patients, were classified according to the aggregation patterns described by De Vos. Eight patients (80%) had particles of type 2. A reevaluation our proceeding data on Delta hepatitis demonstrated that no particles of type 2 were present. These results support the hypothesis that only type 2 particles are markers of non-A, non-B hepatitis.

[The clinical, histological and ultrastructural findings in a case of acute steatosis of pregnancy with a severe cholestatic syndrome]. / Rilievi clinici, istologici e ultrastrutturali in un caso di steatosi acuta gravidica con grave sindrome colestatica.

A case of acute steatosis in pregnancy featuring unusually severe cholestasis is discussed with details of the successful outcome for mother and foetus. The correspondence between clinical picture, histological and ultrasound findings is reported together with a detailed review of the latest reports in the literature and what they reveal about the true incidence of the disease, the variability of the clinical picture, whether and when it is better for both mother and child to interrupt.

Detection of hepatitis C virus-RNA by polymerase chain reaction in dental surgeries.

The mean prevalence of anti-hepatitis C virus (HCV) in Italy is 0.87%. It reaches 2% in Campania, Southern Italy. Approximately 50% of community acquired non-A, non-B (NANB) hepatitis cannot be associated with known parenteral exposure. A recent Italian study has shown that the only demonstrable risk factor in 9% of acute C/NANB hepatitis is dental treatment. There are no data on direct contamination by HCV of dental surgeries. Possible environmental contamination by HCV-RNA was investigated in dental surgeries after treatment of anti-HCV and HCV-RNA positive patients. Thirty-five anti-HCV and HCV-RNA positive patients with chronic hepatitis underwent dental treatment and were enrolled in this study. Eight had chronic persistent hepatitis (CPH), 23 chronic active hepatitis (CAH), and 4 cirrhosis. A total of 328 samples collected from instruments and surfaces were tested after dental treatment of 35 anti-HCV positive patients. The presence of HCV-RNA was determined by polymerase chain reaction (PCR) to evaluate contamination of instruments and surfaces in dental surgeries. Twenty (6.1%) out of 328 collected samples were positive for HCV-RNA. The positive samples were from work benches (two), air turbine handpieces (one), holders (four), suction units (one), forceps (four), dental mirrors (two), and burs (six). Our data indicate that there is extensive contamination by HCV of dental surgeries after treatment of anti-HCV patients and that if sterilisation and disinfection are inadequate there is the possible risk of transmission to susceptible individuals.

HCV-RNA survival as detected by PCR in the environment.

We have shown that HCV-RNA is resistant to drying at room temperature for at least 48 hours. This is a factor which could influence the diffusion of the virus in the general population, which epidemiological studies have shown to be surprisingly high. It should be considered in evaluating the importance of the inapparent parenteral routes of transmission.

Mycelium-yeast transition in Histoplasma capsulatum. Early ultrastructural changes.

The ultrastructural changes which occur during the first 24 h of mycelium to yeast transition have been studied in the dimorphic fungus Histoplasma capsulatum. A temperature shift controls mycelial to yeast transition. During the first 24 h respiratory rate, ATP and cytochrome concentration fall to very low levels. Ultrastructural observations showed that the plasma membrane became undulated and the cell wall lost its characteristic fibrous outer layer. At 8 h the ordered lamellar structure of the mitochondria was no longer apparent. 24 h after the temperature shift 70% of the cells were lysed. The remaining cells contained many cytoplasmic membrane structures; mitochondria were rarely observed. These changes are considered to be the morphological expression of the physiological events characteristic of stage one in mycelial to yeast transition.

Aetiological association of virus like particles with hepatitis C.

Isolation of virus-like particles from sera of anti-HCV positive patients and their ultrastructural characterization are reported. Particles were identified in sera of 5 out of 6 patients tested. Immunoelectron microscopy assay revealed small aggregates of viral particles. Size and morphological criteria suggest that these particles can be classified as Togaviridae.

Prevalence of circulating immunecomplexes and variations of complement fractions in HBV, HDV and HCV infections: statistical analysis and clinical correlations.

Increased levels of circulating immunecomplexes (CIC) have been demonstrated in the serum of patients with HBV infection and HDV superinfection. This finding appears to be correlated to the disease's activity. In this report serum levels of two fractions of CIC (CIC-Clq and CIC-C3d) were evaluated by ELISA method in a sample of 110 subjects with hepatitis infection (HBV, HCV, HDV). Reference values were obtained in a group of 45 healthy subjects (blood donors). Both the CIC fractions were increased in the patients with HCV infection. The most significant increase for both CIC-C1q and CIC-C3d was found in the cirrhotic patients. The complement fractions C3c and C4 were determined in the serum of these patients to investigate a potential pathogenic role of such immunecomplexes. C3c and C4 fractions showed a significant decrease only in the cirrhotic patients, without correlation with the viral agent. Serum levels of C1q complement fraction were not significantly decreased, thus excluding an impaired synthesis of complement fractions. No significant correlation was found between CIC and C3c and C4 fractions in patients with increased levels of CIC, except a slightly significant correlation between reduction of C3c and increase of CIC-C1q. These data suggest a pathogenic action of immunecomplexes in the course of HCV, particularly in the cirrhotic stage.

Presence of papova-like viral particles in cerebrospinal fluid of AIDS patients with progressive multifocal leukoencephalopathy. An additional test for "in vivo" diagnosis.

An "in vivo" diagnosis of progressive multifocal leukoencephalopathy (PML), a neurological opportunistic viral infection in AIDS patients, can be made only by brain biopsy. In order to identify viral particles, we examined the cerebrospinal fluid (CSF) of 15 AIDS patients with focal neurological signs by electron microscopy using negative staining technique. In 2 out of 3 patients with clinical and neuroradiological presumptive diagnosis of PML, the CSF examination revealed papova-like viral particles. Our results support the hypothesis that the severe cell-mediated immunodeficiency reactivates papovavirus from a latent state in the brain, leading to PML. Therefore, the CSF study by negative staining might be a useful test for an "in vivo" diagnosis of PML.

[Ultrastructural study of the mycelium-yeast transition in Histoplasma capsulatum. I. Changes at 37 degrees C]. / Studio ultrastrutturale sulla transizione micelio-lievito in histoplasma capsulatum: I. Cambiamenti a 37 gradi C.

Ultrastructural changes observed during the first 24 hours of mycelium to yeast transition in the dimorphic fungus Histoplasma capsulatum are reported. During this period the plasma membrane becomes undulated and the cell wall loses its characteristic fibrous outer layer. At 8 h the ordered lamellar structure of the mitochondria is no longer apparent. 24 h after the temperature shift 70% of the cells are lysed. The remaining cells contain many cytoplasmic membrane structures; mitochondria are rarely observed. These morphological changes are probably correlated with the physiological events characteristic of mycelial to yeast transition.

[Ultrastructural study of the yeast-mycelium transition in Histoplasma capsulatum. II. Changes at 34 degrees C]. / Studio ultrastrutturale sulla transizione micelio-lievito in Histoplasma capsulatum: II. Cambiamenti a 34 gradi C.

The ultrastructural changes which occur during the mycelium to yeast transition in Histoplasma capsulatum induced by a temperature shift from 25 degrees C to 34 degrees C are described and compared to those observed after a temperature shift from 25 degrees C to 37 degrees C. 24 hours after the temperature shift to 34 degrees C only 8% of the cells are lysed. However, many mitochondria have lost their characteristic elongated form and have become rounded. Vesicular cristae which are no longer oriented parallel to the long axis of the mitochondria are also observed. In contrast a temperature shift from 25 degrees C to 37 degrees C induces lysis of 70% of the cells; mitochondria are rarely observed in the remaining cells. These ultrastructural changes can be correlated with the uncoupling of oxidative phosphorylation and the production of heat shock proteins.

The effect of temperature on the ultrastructure of Histoplasma capsulatum during the mycelium-yeast transition.

The ultrastructural morphology of the early phases of mycelium-yeast transition in Histoplasma capsulatum after a temperature shift from 25 degrees C to only 34 degrees C is described. Under this condition of lower temperature oxidative phosphorylation is not completely uncoupled and maximum production of heat shock proteins (hsp) occurs. 24 h after temperature shift more than 90% of the cells still appear vital. Alterations in the organization of the mitochondrial cristae are the only ultrastructural changes observed in these cells. In contrast, 70% of the cells degenerate 24 h after a temperature shift from 25 degrees C to 37 degrees C and in the remaining cells mitochondria are rarely observed. These observations are discussed in relation to the production of hsp, the uncoupling of oxidative phosphorylation and the virulence of different strains of H. capsulatum.

Prevalence of intranuclear particles in liver pathologies.

An ultrastructural study of the prevalence of electron dense 23-27 nm intranuclear particles was carried out on liver biopsies from patients with NANB chronic active hepatitis (CAH), Delta + CAH, HBsAg + CAH, nonviral liver pathologies and in one healthy volunteer. The particles were classified according to aggregation pattern and were found to be correlated with NANB CAH and Delta + CAH. No particles were observed in nonviral liver pathologies. A close antigenic relationship has been shown between the cytoplasmic alterations observed in NANB and delta hepatitis in chimpanzees. Our data indicate that there is a structural similarity between the intranuclear particles seen in both Delta and NANB hepatitis, thus reinforcing the hypothesis that the NANB and Delta agents are closely related.

Prevalence of hepatitis C virus genotypes in southern Italy.

HCV is ubiquitous. In 50% of all cases it causes chronic hepatitis that often evolves into liver cirrhosis and hepatocellular carcinoma. Recently HCV has been classified in 5 genotypes by Okamoto. The purpose of this study is to evaluate the prevalence of 5 genotypes in Campania, a region of southern Italy, where the prevalence of anti-HCV antibodies ranges from 0.87 to 4%, and to evaluate the correlation between the HCV genotypes and the severity of histological damage. One-hundred- and-thirty-five anti-HCV positive patients were enrolled and tested by PCR to identify HCV-RNA. One-hundred-and-twenty-four patients resulted HCV-RNA positive. Genotyping was performed as described by Okamoto et al. with minor modifications of the specific primer to type III proposed by Silini et al. Eight patients were negative for all genotypes. Eight patients were positive for type I(1a), 61 for type II(1b), 39 for type III(2a), 11 for type IV(2b) and 1 for type V(3a). In 4 cases two different genotypes were present in the same sample [II(1b)-IV(2b), III(2a)-II(1b) twice, III (2a)-IV(2b)]. Histological evaluation of liver damage showed: CPH (22 cases), minimal CAH (56), severe CAH (31) and liver cirrhosis (15). There was no statistically significant correlation between the 5 genotypes and the severity of histological damage. Data on the prevalence of genotype II (1b) in Italy are similar to those reported for other European countries. The prevalence of genotypes in southern Italy is similar to that reported in the population of northern Italy.

Ultrastructural effects of PCMS on mycelial-yeast transition in Histoplasma capsulatum.

The Yeast phase of Histoplasma capsulatum has stringent growth requirements. Transition from mycelium to yeast takes place only in the presence of cysteine and can be blocked by the -SH groups inhibitor p-chloromercury-phenylsulfonic acid (PCMS). Ultrastructural studies show lysis and degeneration of PCMS treated mycelium grown at 37 degrees C for 24 hours. Only 50% of PCMS treated mycelium appear degenerate when grown at 34 degrees C for 24 hours. The remaining cells have normal morphology with only slight changes in the cell wall structure. The effect of PCMS is permanent and hereditary. Mice injected with PCMS treated mycelium do not develop disease and are resistant to virulent strains of H. capsulatum when challenged.