Detection of hepatitis B virus in the liver by in situ hybridization (ISH) in HBsAg seropositive and seronegative patients.

The presence of hepatitis B virus (HBV) DNA in the liver of 119 patients was studied to assess the diagnostic value of in situ hybridization (ISH) and its relationship with viral replication and histological liver damage. Liver biopsies of 119 patients (55 hepatitis B surface antigen -HBsAg- seropositive and 64 HBsAg seronegative) were studied retrospectively. Among the HBsAg seropositive patients, the ISH was positive in 26 cases (47%) and negative in 29 (53%) and the former group had higher levels of serum transaminases. The hepatocyte number with positivity for HBsAg and hepatitis B core antigen (HBcAg) in the liver were similar in both ISH-positive and -negative patients. The histological activity index (Knodell) was higher in ISH-positive patients (11 vs 7, p < 0.001). Six patients out of 12 were positive by PCR. In the HBsAg seronegative patients, the ISH was negative in 57 cases and positive in 7. These 7 were positive for anti-HBs (5 cases) and/or anti-HBc (6 cases); 4 were confirmed by PCR. Thus, our data suggest that the ISH technique is useful for detecting viral nucleic acid in the liver, but that the HBV-DNA cannot always be considered as a replication marker, because we also show that some HBsAg seronegative patients with chronic liver disease do have HBV-DNA in their liver cells.

Histological and immunohistochemical assessment of liver biopsies in morbidly obese patients.

AIMS: To study liver lesions in morbidly obese patients who underwent liver biopsy at the time of bariatric surgery to define histological lesions, especially inflammatory infiltrate, diagnostic categories and the possible influence of gender in this respect. METHODS AND RESULTS: 110 biopsies (36 males-M- and 76 females -F-) were evaluated and categorised, according to the NAS (NAFLD -non alcoholic fatty liver disease- Activity Score) system and other criteria, as non-NAFLD (15.5%, F predominance), non-alcoholic steatohepatitis (NASH) (16.5%, M predominance), non-alcoholic hepatosteatosis (NAHS) (21%, F predominance) and, the most numerous group, NASH-borderline (NASH-BORD) (47%), with three subgroups, characterised by centrozonal lesions, portal area preferential involvement or affecting both areas. The predominant form of hepatocytesteatosis was mixed with a multivesicular component that was present in most cases with fibroinflammatory portal involvement. Nuclear glycogenosomes were found in greater number of biopsies in patients in the third and sixth decades. Portal inflammation was present in a large number of cases (M predominance); the application of immunohistochemical techniques (myeloperoxidase and CD68 antibodies) to evaluate lobular inflammation revealed "surgical hepatitis" in one third of the cases, and the presence of microgranulomas (CD68+) (M predominance), which were more abundant with increasing lesion severity. CONCLUSIONS: Portal inflammation and multivesicular hepatocytesteatosis are highly prevalent in morbidly obese patients. This study identifies a new subtype of NASH-BORD characterized by centrizonal and porto-periportal area involvement and the existence of liver biopsies without steatosis. CD68+ microgranulomas constitute an unequivocal marker of lobular inflammation in surgical biopsies and of lesion severity, which is gender-related.

Liver fibrosis assessment with semiquantitative indexes and image analysis quantification in sustained-responder and non-responder interferon-treated patients with chronic hepatitis C.

BACKGROUND/AIMS: The effect of interferon on the reduction of liver fibrosis is controversial. We aimed to compare semiquantitative methods with a quantitative digital image analysis system to assess liver fibrosis in biopsies from patients with chronic hepatitis and different responses to interferon. METHODS: We studied 98 liver biopsies with chronic hepatitis C before and after recombinant interferon alfa-2 treatment, using conventional histological assessment, grading of histological activity, scoring/staging of fibrosis (Knodell and Scheuer), and quantification of fibrosis with image analysis (FibroQuant). RESULTS: Sustained-responders to interferon showed a significant reduction in histological lesions and in their Knodell and Scheuer activity indexes. The semiquantitative systems showed no reduction in fibrosis. The FibroQuant application showed a significant reduction in porto-periportal and septal areas among sustained-responders (P < 0.001) and non-responders (P < 0.05), and in porto-periportal and septal fibrosis areas only in sustained-responders (P < 0.001), whereas the percentage of fibrosis increased in non-responders (P < 0.001). CONCLUSIONS: The Scheuer system is useful for the daily evaluation of fibrosis, but the FibroQuant application provides more objective data on the anti-fibrogenic effects of interferon, which include a reduction in the porto-periportal area in sustained-responders and non-responders, accompanied by a reduction in the area of fibrosis only when the viral replication has ceased.

[Identification of cell types in periodontal pockets with scanning electron microscopy]. / Identificación de tipos celulares en la bolsa periodontal con microscopio electrónico de barrido.

Sixteen teeth from patients with advanced periodontal disease were processed with the direct fixative technique for observation with a Scanning Electron Microscope in order to test the adequacy of this technique and to observe the components of the tooth wall of the periodontal pocket. Different cell populations are described emphasizing the appearance of intact neutrophils. The results suggest that this fixative method may be an improvement on the conventional techniques for SEM.

Monosomy 22 with humoral immunodeficiency: is there an immunoglobulin chain deficit?

The cytogenetic analysis of a patient with selective deficit of IgA and decrease in IgM, IgE, and IgG is presented. Using trypsin-Giemsa banding the karyotype showed monosomy 22 (45,XX,-22). The interest of this case lies in the rarity of the illness and in the association of monosomy 22 with hypogammaglobulinaemia and selective deficit of IgA, particularly as this chromosome is known to contain genes coding for immunoglobulin chains.