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Biomedical subjects

V A Alves

Publications and source records attributed to V A Alves.

At least 19 recordsLinked to original sources

Spontaneous hepatitis B surface antigen clearance in a long-term follow-up study of patients with chronic type B hepatitis. Lack of correlation with hepatitis C and D virus superinfection.

We investigated the frequency of HBsAg clearance and the possible role of viral superinfection in a long-term follow-up of 184 patients with chronic hepatitis B (CHB). Our subjects were 184 patients with chronic hepatitis B and the follow-up was 12-216 months (mean 66.2 +/- 53.7 months). The investigative methods used were: immunoenzymatic assays for HBV, HCV, HDV, and HIV markers; polymerase chain reaction (PCR) for HBV DNA; and liver biopsy and immunoperoxidase. During the follow-up, 20 of the 184 patients cleared serum HBsAg. A comparison of patients with persistent HBsAg(group I) and of those who cleared this marker (group II) showed a significant difference in mortality (P = 0.002) between the two groups and a tendency to a more severe exacerbation (flare) in group II (P = 0.07). Antibodies to hepatitis C and D virus as well as antibodies to HIV were equally distributed in both groups. Thirteen patients (7.9%) from group I, but none from group II, subsequently developed hepatocellular carcinoma. These results suggest that the frequency of spontaneous clearance of HBsAg during chronic HBV infection is low. No determinant factor for the clearance was found, including the presence of liver cirrhosis. Serum HBV DNA was undetectable by PCR after clearance in 16 out of 17 patients.

Adult

Demonstration of mycobacterial antigens in skin biopsies fron suspected leprosy cases in the absence of bacilli.

Skin-biopsies from fifty-six patients suspected of early leprosy from Bahia State, Brazil, were examined histopathologically. The Fite-Faraco staining failed to demonstrate acid-fast bacilli in this material. The prominent features of the lesions were inflammation of the neurovascular bundles and sometimes inflammation of the skin appendages. The non-specific infiltrate was predominantly composed of histiocytes and lymphocytes. In 41 cases (73.2%) epidermal atrophy was also present. The avidin-biotin peroxidase technique was used with primary antibodies to detect bacillary antigens (anti-BCG serum) and nerve branches (anti-S-100 protein serum). Immunohistochemical detection of bacillary antigens using the anti-BCG serum was positive in 28 cases (50%). A positive staining for S-100 protein was observed in 40 cases (71.4%) in dendritic antigen-presenting cells of the skin. The detection of bacillary antigens, together with the clear demonstration of nerve bundles enhanced our capacity to fulfill morphologic criteria for the diagnosis of early leprosy. Our observations indicate that the use of immunohistochemical methods represent a useful tool for the early diagnosis of leprosy.

Adolescent

Frequency of epithelial giant cells in smears with Papillomavirus.

We re-screened cytological smears from patients who presented Human Papillomavirus (HPV) infection detected morphologically in order to verify the frequency of associated giant epithelial cells, as recently mentioned in the literature. Our results demonstrated that 14 (13.7%) of 102 cases showed the giant epithelial cells, associated or not with macronmucleosis, bi or multinucleation. Epithelial giant cells showed a poor reaction to the avidin-biotin-peroxidase to the BPV-1 antigen. We conclude that epithelial giant cell is not a frequent cytologic feature in HPV, and the avidin-biotin-peroxidase study did not detect a marked expression of HPV late antigen.

Carcinoma in Situ

Immunohistochemistry of medullary thyroid carcinoma and C-cell hyperplasia by an affinity-purified anti-human calcitonin antiserum.

BACKGROUND: The diagnosis of medullary thyroid carcinoma (MTC) depends on the calcitonin immunohistochemistry. Familial MTC is associated with C-cell hyperplasia (CCH), whereas sporadic MTC is not. A specific and sensitive calcitonin immunohistochemistry is necessary for the diagnosis of MTC and CCH. METHODS: An affinity-purified anti-calcitonin antiserum (APxCT) was used for immunohistochemistry of the thyroids of 15 patients with MTC. The thyroids of five patients with familial MTC were studied in detail, with each gland sectioned in 48 areas. RESULTS: Between three and ten independent MTC were found in each thyroid, and CCH was found in all five patients (24.2%, varying from 8.4-56.3% of the 48 areas from each thyroid). MTC and CCH were localized mainly in the middle third and in the central axis of the thyroid lobes. They often were found together in the same area (in a total of 21 areas for the five thyroids sectioned in 48 areas) but ten areas with MTC did not have CCH, and 37 areas with CCH did not have MTC. In ten thyroids partially studied, CCH was indicated in three patients thought to have sporadic MTC. In two thyroids, with follicular and papillary carcinoma, a higher density of C-cells was found around the tumors, but disease was not characterized as CCH. CONCLUSIONS: APxCT antiserum increased the immunohistochemical specificity and sensitivity. The distinction of the familial from the sporadic MTC requires a careful and extensive search of CCH. C-cells in high density may be found around follicular cell carcinomas, being a potential source of diagnostic error.

Adolescent

Human fatal yellow fever. Immunohistochemical detection of viral antigens in the liver, kidney and heart.

An immunohistochemical method to detect yellow fever antigen was developed using immune sera from rabbits and hamsters and hyperimmune ascitic fluid from mice. A search for the antigen was carried out in liver, kidney and heart in three fatal cases of yellow fever. In the liver it was present in the cytoplasm of hepatocytes, Councilman bodies and Kupffer cells. Yellow fever antigen was also detected in renal tubular epithelium and in groups of myocardial fibers. These findings suggest that viral replication occurs at sites other than the liver. Since yellow fever shares many features with other haemorrhagic fevers the use of immunohistochemistry can impart a significant improvement in the accuracy of its histopathological diagnosis.

Adult

Leptospiral antigens in the liver of experimentally infected guinea pig and their relation to the morphogenesis of liver damage.

In order to investigate the morphogenes of experimental leptospirosis by morphologic and immunohistologic methods, 24 guinea-pigs were inoculated intraperitoneally with L. interrogans serogroup Icterohaemorrhagiae. They were divided in 6 groups, sacrificed from the 1st to the 6th day of infection. Semiquantitative analyses of histopathological liver lesions were performed in 1 micron sections of tissue embedded in glycol-methacrylate. The distribution of leptospiral antigen (L. Ag) and its glycolipoprotein (GLP) was demonstrated by peroxidase-antiperoxidase on paraffin embedded tissue. Significant lesions appeared at the 4th day of infection, progressing to a peak on the 6th day. Inflammation was associated with injury of the portal triad. Liver cells showed either swelling or acidophilic degeneration and necrosis, together with loss of cell cohesion, leading to disarray of liver cell plates. Mitochondria were found progressively enlarged and irregularly distributed. L. Ag expression was parallel to the morphological changes. Portal distribution was significant at the 4th day and on later stages centrilobular localization became predominant. Spiral forms suggestive of intact leptospires were initially found but, chiefly at the 6th day, L. Ag was seen in granules, probably resulting from phagocytosis. GLP staining was similar to granular L. Ag in morphology, and distribution. Cytokeratin condensation was seen in liver cells with acidophilic necrosis and was marked in areas of disorganization of cell plates. Our findings lead us to hypothesize a direct leptospiral cytotoxic effect on endothelial and on liver-cell membranes. At first, leptospires themselves would induce subcellular changes acting mainly on membrane permeability. Afterwards, their granular forms, including GLP, would act as adjuvant factors. These findings demonstrate that the disarray of liver cell plates at the late phase of the disease is genuine.

Animals

Detection of leptospiral antigen (L. interrogans serovar copenhageni serogroup Icterohaemorrhagiae) by immunoelectron microscopy in the liver and kidney of experimentally infected guinea-pigs.

Guinea-pigs were experimentally infected with L. interrogans serovar copenhageni serogroup Icterohaemorrhagiae and their liver and kidney were studied by immunoelectron microscopy using the post embedding indirect immunogold labelling technique. Primary antibody was a purified rabbit anti-serum produced against the same leptospiral strain used in the inoculum. Gold-labelled leptospiral antigen (LAg) was found close to cell membranes of hepatocytes, kidney tubular cells and endothelial cells of the interstitial capillaries of the kidney. Afterwards it was internalized by hepatic and tubular cells, and eventually found in lysosomes. Phagolysosomes of Kupffer cells were also found to contain remnants of degraded leptospires and gold-labelled LAg. Gold-labelled intact leptospires were detected at the enlarged intercellular spaces between hepatocytes at the areas of hepatic cell plate disarray, showing the potential for leptospiral migration during the septicaemic phase of the disease potentially contributing to the pathogenesis of the lesions. The affinity of leptospiral antigenic material for cell membranes suggests an initial interaction with cell surface proteins followed by its internalization and cell damage. The nature of antigenic material detected, however, remains undefined; it may be a toxin, an enzyme or any other factor/s involved in leptospiral virulence.

Animals

Restaging of colorectal cancer based on the identification of lymph node micrometastases through immunoperoxidase staining of CEA and cytokeratins.

The present study was performed to identify tumor cells in lymph nodes from colorectal adenocarcinomas considered free of disease by the classic hematoxylin-eosin stain, based on the detection of the carcinoembryonic antigen (CEA) and cytokeratins in neoplastic epithelial cells. For this purpose, 603 lymph nodes from 46 lesions were stained by the peroxidase-antiperoxidase technique. Tumor cells were detected in 22 nodes from 12 patients, mainly in the subcapsular sinuses, permitting a restaging of these patients into two groups: those now considered to have metastatic disease and those free of metastases. However, the 5-year follow-up showed no statistical differences in survival between the two groups.

Adenocarcinoma

Leptospiral antigens (L. interrogans serogroup ictero-haemorrhagiae) in the kidney of experimentally infected guinea pigs and their relation to the pathogenesis of the renal injury.

The search for leptospiral antigens (L. interrogans serogroup icterohaemorrhagiae) was carried out in 24 guinea pigs experimentally inoculated with 1 ml of culture containing 10(7)-10(8) leprospires and sequentially sacrificed from the first until the 6th day of infection. Semiquantitative analysis of histopathological variables comprising kidney interstitium, tubules and glomeruli was done in 1 micron sections of tissue embedded in glycolmetacrylate. Leptospiral antigen (LAg) and its glycolipoprotein (GLP) expression were detected through PAP in paraffin embedded tissue. The mild interstitial involvement of the kidney, manifested chiefly by oedema and focal interstitial nephritis seen at the 4th day, progressed to tubular damage at the 6th day, characterized by either swelling or cytoplasmic acidophilia of epithelial cells with loss of cell cohesion and sloughing of cells into the tubular lumina. Brush border alterations and mitochondrial changes were observed. Endothelial cell injury was noted in the interstitial vessels. LAg expression was parallel to the kidney changes: small deposits of elongated forms of LAg were detected at the 4th day either within the vascular lumen or free in the interstitium. A rise in the antigen expression was observed at the 5th day when it was seen either around tubules or in their walls. LAg was detected inside the tubular lumina at the 6th day of infection when granular LAg and GLP were abundant. This sequence reproduces the pathway of leptospires in the kidney and the crescent amounts of antigens detected toward the end of the experiment, with antigen concentration in cases of major tissue damage suggesting a direct action of the microorganisms and/or their products in the pathogesis of the lesions.

Animals

Use of monoclonal antibodies against human T cells for clinical diagnosis by immunohistochemical procedures.

Monoclonal antibodies (Mabs) were produced against human T cell membrane antigens. Sixteen Mabs were studied and six were selected for immunohistochemical assays on paraffin-embedded tonsil sections. Two Mabs (2D7 and 1E2) specifically recognized T-lymphocyte areas in sections of pathological tissues originating from lymphoproliferative diseases, and reacted with proteins of approximately 80 kDa. Most of the Mabs produced thus far are only suitable for immunohistochemical assays on frozen section. Only a few Mabs recognize lymphoid markers on paraffin-embedded sections, a procedure which permits a more extensive and practical application of Mabs in clinical diagnosis. These antibodies should be valuable in diagnosing T cell-related diseases and their large scale production should reduce laboratory costs because all reagents currently available are imported.

Antibodies, Monoclonal

Intrahepatic bile duct changes in human hepatosplenic schistosomiasis mansoni.

Wedge liver biopsies of 132 patients with hepatosplenic mansonian schistosomiasis were studied and divided in two groups according to the presence (Group I - 69 cases) or absence (Group II - 63 cases) of markers of the actual presence of the parasite in the liver tissue. Histological variables indicating bile duct injury were analysed in each case: periductal fibrosis, hyperplasia of the bile duct epithelium, bile duct degeneration, and marginal ductular proliferation. The presence of one or more of these variables defined two sub-groups: A - bile duct lesions present (73 cases), and B - bile duct lesions absent (59 cases). The variables "bile duct degeneration" and "ductular proliferation" were related to the actual presence of the parasite in the host. In 55.3% of all cases of human mansonian schistosomiasis a spectrum of injuries to the bile ducts was present. Epithelial hyperplasia alone or associated with patterns of mucopolysaccharide production was observed in 87.6% cases of the Sub-group A. The bile ducts changes in mansonian schistosomiasis are close to those described in liver fluke infestations such as clonorchiasis, fascioliasis and opistorchiasis. Statistical analysis revealed that high mucopolysaccharide production was associated with epithelial hyperplasia. The pathogenesis of the bile duct changes in human mansonian schistosomiasis and its relation to the parasitic infestations and their antigens is discussed.

Bile Ducts, Intrahepatic

Myocardial infarct due to a unique atrial myxoma with epithelial-like cells and systemic metastases.

A 73-year-old man had myocardial infarct and coronary emboli from a left atrial myxoma diagnosed at necropsy. The tumor was attached to the atrial roof and showed no local myocardial infiltration. Transmission electron microscopic (TEM), light microscopic, and immunoperoxidase (IPX) studies confirmed the neoplastic character of this lesion and pointed to undifferentiated mesenchymal cells as the origin of the atrial myxoma cells. This case emphasizes two aspects: (1) glandlike structures were also found in the myxoma, and their epithelial-like nature was supported by TEM and IPX studies, which showed positivity for carcinoembryonic antigen and H blood substance; (2) many systemic tumor masses were found, and their metastatic nature was evidenced by the markedly infiltrative and destructive character; the only cytologic marker that could discriminate this case from other usual, noninfiltrative cardiac myxomas was the epithelial-like cells.

Aged

Detection of leptospiral antigen in the human liver and kidney using an immunoperoxidase staining procedure.

Detection of leptospiral antigen using an immunoperoxidase staining (IP) procedure was carried out on fifteen samples of human liver, (nine from autopsies and six from biopsies) and nine samples of human kidneys (eight autopsies and one biopsy). The IP staining procedure to detect leptospiral antigen (L. interrogans serovar icterohaemorrhagiae) in human liver and kidney proved to be a reproducible method useful on paraffin embedded tissues after formalin, Bouin's or Helly's fluid fixation. Furthermore, the IP procedure on paraffin-embedded tissue appears to have potential as an aid to the diagnosis. IP stained leptospiral antigen was detected in portal spaces of the liver, engulfed by cells of the mononuclear phagocyte system, in the interstitium of the kidney, and lining vessel walls of both liver and kidney. The results suggest that in acute human leptospirosis (L. interrogans serovar icterohaemorrhagiae) the main factors in the pathogenesis of the lesions are related to the presence of organisms and/or their virulence, including products released by lysis.

Adult

Unexpected low prevalence of delta antibodies in the east Amazon region and São Paulo: evidence for regional differences in the epidemiology of delta hepatitis virus within Brazil.

Antibodies (anti-HD) to hepatitis delta virus (HDV) were tested by radioimmunoassay in 207 human serum samples from the eastern Amazon (states of Pará and Amapá) and São Paulo, Brazil. 42 Amazon HBsAg asymptomatic carriers were negative for anti-HD. 84 São Paulo HBsAg asymptomatic carriers were also negative. Among the 81 HBsAg patients from São Paulo with different liver diseases, only one had anti-HD. Liver biopsy of this chronic active hepatitis case was positive for HBsAg, HBcAg and HDAg in liver, by an immunoperoxidase technique. The low prevalence of HDV infections in São Paulo and eastern Amazon was unexpected and contrasts with the recent reports of high prevalence in the western Amazon region. Such regional differences emphasize the need for extensive and precise worldwide epidemiological studies of HDV.

Adult

Cardiovascular involvement in human and experimental leptospirosis: pathologic findings and immunohistochemical detection of leptospiral antigen.

Twenty hearts from patients dying of leptospirosis were studied. Interstitial myocarditis was found in 50% of the cases, and a significant statistical correlation was observed between myocarditis and the inflammatory involvement of the conduction tissue. Acute coronary arteritis, affecting the main branches of the coronary arteries, was observed in 70% of the cases, and this finding also correlates significantly with interstitial myocarditis. Aortitis was found in 57.8% of the cases. When serum against L. interrogans serovar icterohaemorrhagiae was used, focal IP antigen deposits were observed in the coronary arteries and in the aorta. Experimental data from 12 guinea-pigs inoculated with L. interrogans serovar icterohaemorrhagiae showed a focal myocarditis involving mainly the subendocardial and pericoronary heart tissue, with IP antigen deposits in the same sites. Leptospirosis might be visualized as a generalized illness resembling other infectious vasculitides. The heart and main vessels are involved during the septicaemic phase of the disease, and bacterial migration, toxin(s), enzymes and/or antigenic products liberated by bacterial lysis might account for the increased endothelial permeability with antigen deposits and inflammation.

Adult