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Biomedical subjects

N C Nayak

Publications and source records attributed to N C Nayak.

At least 19 recordsLinked to original sources

Immunohistochemical detection of serum autoantibodies--a new technique using routine formalin fixed paraffin sections.

Detection of autoantibodies in serum is important in the diagnosis of primary immune mediated diseases. Tests of these antibodies are conventionally done by indirect immunoflourescence (IIF) on frozen sections of fresh target tissues from the murine species. More recently enzyme-linked immunoassays and IIF on cultured human epithelial cells or on fresh frozen sections of murine tissues coated onto wells or glass slides are also being used. But these tests are more expensive and generally not easily available to laboratories in developing countries. Obtaining small animals for preparing frozen sections of fresh tissue is also getting to be increasingly difficult. A simple; new and inexpensive technique was developed to perform IIF on routine paraffin sections of human tissues following antigen unmasking. This, technique offers qualitatively good, consistent, species specific and dependable results with several advantages over the conventional IIF on animal tissue frozen sections, particularly in a sausage block made from different types of tissues. Immunoperoxidase stain for autoantibodies can also be easily performed with the advantages of permanent preservation and clearer evaluation in light microscopy. Most importantly the technique is easily affordable and practicable in all histopathology laboratories.

Animals↗

Hepatocellular carcinoma--a model of human cancer: clinico-pathological features, etiology and pathogenesis.

Hepatocellular carcinoma (HCC), a unique human neoplasm, has interested many in several fields of biological and health sciences. This cancer is credited as the first that can be largely eliminated by a safe anti-viral vaccine and other transmission control measures. It is also the first cancer for which a reliable diagnostic tumour marker was identified and studies on this tumor in humans and animals have provided a large body of information on causation and step-wise evolution of cancers. Being a common and rapidly fatal tumour, mainly affecting males in the more populous developing countries, HCC may well be the commonest cancer of the human male. Clinical features are not specific for HCC but manifestations represent varying combinations of those due to cirrhosis which is a very frequently associated and pre-existent disease, those due to tumour and those due to malignancy. This tumour commonly takes a massive form with satellite nodules or a scattered multinodular form. A fibrolamellar variant is biologically and clinically quite different from the usual HCC and the small capsulated variant is seen in some geographic areas. Microscopically the trabecular variety is common and differentiation from metastatic cancers and benign lesions may need use of newly described immunohistochemical markers in addition to clinical evidence of cirrhosis. Hepatitis B and C viruses, dietary aflatoxin B1 and cirrhosis from any cause are common risk factors in that order of importance. Several lines of evidence including molecular biology and animal studies support these etiological linkages. Studies in experimental animals using chemical carcinogens, different hepatotropic viruses and aflatoxin have greatly helped our understanding of carcinogenesis in general and hepatocarcinogenesis in particular. Individual HCC risk factors may contribute to summation of mutations subsequent to repeated and continued liver cell injury act as carcinogen/co-carcinogen or be involved in both capacities.

Aflatoxin B1↗

A multicentric study with arteether in patients of uncomplicated falciparum malaria.

Two hundred and sixty seven patients of uncomplicated P. falciparum malaria completed study in a multicentric phase III clinical trial of Arteether. Arteether was given intramuscularly in a dose of 150 mg daily for three consecutive days. Each patient was followed upto 28 days of alpha, beta arteether therapy. The cure rate was 97% with fever clearance time between 1-7 days (24-168 hours) and parasite clearance time between 1-3 days (24-72 hours). Parasite reappearance rate was found to be 3% and reported at only three of the centres. Following the treatment no adverse effect was observed on haematological, biochemical and vital clinical parameters.

Adolescent↗

Multicentric clinical trials for safety and efficacy evaluation of alpha;beta arteether in complicated P. falciparum malaria.

OBJECTIVE: To evaluate efficacy of alpha;beta arteether in patients of P. falciparum malaria presenting with complications was undertaken in a multicentric clinical trial. METHOD: Each patient who consented to undergo clinical trial with parenteral Arteether was treated with a fixed dose schedule of Arteether given intramuscularly in a dose of 150 mg once a day on three consecutive days. Every patient was followed upto 28 days with clinical, haematological and parasitological monitoring every day upto one week and thereafter at 14, 21 and 28 days. The response was assessed in terms of fever clearance time, parasite clearance time, cure rate and parasite reappearance rate. RESULTS: A total of 211 patients of P. falciparum malaria were included in the study from four centres (Bhilai, Guwahati, Jamshedpur and Rourkela). Results of this study showed that fever clearance time ranged between 24-168 hours, parasite clearance time ranged between 24-120 hours and overall mortality ranged between 4-8.5%. Out of 211, only 14 patients expired during the study, of these, 10 patients expired within first two days i.e. before completing the three day schedule of arteether therapy. Tolerability to arteether injection was good in all these patients and no untoward effects were experienced or reported during the study. Overall cure rate observed in these studies was 93%. CONCLUSION: This study shows a rapid parasite and fever clearance in patients of complicated P. falciparum malaria.

Adolescent↗

Immunohistochemical detection of hepatitis C virus antigen in paraffin embedded liver biopsies from patients with chronic liver disease.

A simple and reproducible immunohistochemical (IHC) staining method that adequately identifies and localizes hepatitis C virus (HCV) in human livers is still not available. We performed IHC staining using both a new monoclonal antibody against HCV and a polyclonal human anti-HCV IgG to 94 liver biopsies from HCV seropositive patients with chronic hepatitis and 15 control liver biopsies. Positive nuclear staining was consistently observed predominantly in hepatocytes and much less in lymphocytes with either antibody in 57% of anti-HCV seropositive cases but in none of the controls. However, the monoclonal antibody yielded a stronger positive reaction and in a greater proportion of hepatocytes. In 78% of the positive cases, more than a quarter of the hepatocytes showed nuclear staining. The degree of hepatic HCV load as revealed by intensity and extent of positive staining did not correlate with histological changes in the liver. The new monoclonal antibody against HCV appeared to be suitable for identifying HCV in tissues by a simple IHC stain and can be used to explore the pathogenesis of liver injury induced by this virus.

Antibodies, Monoclonal↗

Immunohistochemical study of proliferating mesenchymal spindle cells in heterogenous soft tissue lesions.

Proliferation of mesenchymal spindle cells is a main event in a variety of lesions with similar morphological features but widely divergent biological behaviour. In order to identify criteria for precise histological diagnosis, 60 human soft tissue lesions, divided into 40 cases of fibroblastic cell proliferation, 10 smooth muscle cell tumours and 10 nerve sheath cell tumours, were examined for the immunohistochemical profile of the main lesional cell in addition to other histological features. The three groups could be differentiated by determining the lineage of the constituent spindle cell on the pattern of expression of vimentin, alpha-smooth muscle actin (ASMA) and macrophage antigen CD68 (MA-CD68). Smooth muscle cells expressed ASMA and vimentin but not MA-CD68, while nerve sheath cells were negative for ASMA but positive for vimentin and MA-CD68. The fibroblastic cell lesions as a group were easily differentiated on the basis of positive reactivity for all three markers but individual lesions could only be distinguished by additional assessment of histological features. Because of consistent co-expression of ASMA, vimentin and MA-CD68 in the spindle mesenchymal cell present in all varieties of lesions in this heterogeneous group, we suggest that this proliferating "fibroblastic" cell is phenotypically a fibromyohistiocyte.

Actins↗

The nature and significance of liver cell vacuolation following hepatocellular injury--an analysis based on observations on rats rendered tolerant to hepatotoxic damage.

Swelling with nonlipid cytoplasmic vacuolation of diffusely distributed hepatocytes is seen consistently after mild acute and subacute liver injury. Several lines of evidence point to the possibility that this change may reflect a cellular adaptation beneficial to the host, rather than a degenerative change. The nature and significance of this morphological manifestation were tested in batches of albino rats given small doses of a variety of hepatotoxins, some of which were subsequently challenged with a large highly necrogenic dose of carbon tetrachloride (CCl4). Morphological and biochemical investigations showed that cytoplasmic vacuolation of liver cells following low doses of toxins was due to excess accumulation of glycogen, predominantly of the monoparticulate form. These cells lacked features of degeneration or regeneration and were much less susceptible to injury by the large dose CCl4, as assessed by structural and serum enzyme analyses. This tolerance to toxic damage seemed to be associated with excess accumulation of intracellular glycogen. We conclude from these and other observations on animal and human livers that many of the vacuolated hepatocytes seen in liver injury are cells adaptively altered to resist further insult rather than cells undergoing hydropic degeneration, as is commonly believed.

Adaptation, Physiological↗

Indian childhood cirrhosis--like liver disease in an Arab child. A brief report.

An Arab female child presented with rapidly progressive liver disease, with apparent onset in late infancy and death at 15 months. Microscopy showed panacinar hepatitis, portal and pericellular fibrosis, and diffuse Mallory bodies in the absence of steatosis or significant cholestasis. Hepatic copper concentration was moderately elevated. Known causes of early childhood cirrhosis were excluded. This case meets most of the established criteria of Indian childhood cirrhosis, yet is unusual in its occurrence in a child of Arab ancestry and in having a moderate degree of hepatocellular copper overload.

Copper↗

Aflatoxin B1 induced hepatocarcinogenesis in neonatal rats.

Role of cell replication on aflatoxin B1 (AFB1) induced hepatocarcinogenesis was investigated in neonatal rats showing persistence of cell replication in the liver for 21 days of post natal life. Adult (8-10 weeks old) rats displaying no hepatocytic proliferation served as controls. Three doses of AFB1 were administered to both the groups at intervals of 48 hr with the doses starting on 10th day of age in the neonatal group. Appearance of phenotypically altered preneoplastic hepatocytes was quantitated in both the groups. A significantly higher incidence of preneoplastic foci was recorded in neonatal rats as compared to adult animals. The results suggest that presence of cell replication in neonatal rats at the time of AFB1 administration enhances the process of hepatocarcinogenesis.

Aflatoxin B1↗

Comparative evaluation of the immunogenicity of yeast-derived (recombinant) and plasma-derived hepatitis B vaccine in infants.

The immunogenicity of plasma-derived (HB Vax,MSD) and recombinant hepatitis B virus (Engerix B, SK&F) vaccines was evaluated in infants born to hepatitis B virus carrier mothers. The vaccination was carried out at 1 day, 1 month, and 6 months of age using 10 micrograms of the vaccine given intramuscularly. A total of 83/88 (94.3%) and 74/79 (93.6%) of the infants receiving the plasma-derived vaccine and yeast-derived vaccine showed antibody to hepatitis B surface antigen (anti-HBs). None of the maternal factors studied apart from the HBeAg positivity corellated with vaccine failure. The yeast-derived vaccine gives marginally lower antibody titre than the plasma-derived vaccine. The group-specific anti-"a" antibody was less than 10% of the total anti-HBsAg titre. It was observed that the vaccine alone without prior administration of hepatitis B immunoglobulin is effective in perinatal infection.

Biomarkers↗

Significance of natural polymerized albumin and its receptor in hepatitis B infection of hepatocytes.

Lack of information regarding the presence of native albumin polymer in serum and its structural similarity to the one produced by glutaraldehyde treatment casts doubt on the postulate that hepatitis B virus attachment to hepatocytes is mediated through polymerized albumin. We used a sandwich enzyme-linked immunosorbent assay with murine monoclonal antibodies raised against glutaraldehyde-polymerized albumin to detect native albumin polymer in human serum and its cross-reactivity with other albumin polymers. Presence of polymerized albumin receptor on the HepG2 cell was studied by radioreceptor assay. Purified hepatitis B virus and synthetic peptide analogous to part of pre-S2 sequence (120-145) were used to study polymerized albumin-dependent attachment of the virus to HepG2 cells. Antibodies raised against pre-S2 peptide were used to inhibit the pre-S2 and hepatitis B virus attachment to HepG2 cells. Glutaraldehyde-treated polymerized albumin was found to be immunologically cross-reactive with native albumin polymer. Its levels were found to be significantly raised in sera of patients with liver diseases. Polymerized albumin has specific saturable receptor on HepG2 cells with two classes of binding sites of different equilibrium dissociation constant (Kd1 = (16 +/- 9.6)pmol/L and Kd2 = (1,019 +/- 172)pmol/L. Albumin monomer was unable to compete for the polymerized albumin receptor sites on HepG2 cells. Anti-pre-S2 antibodies inhibit hepatitis B virus and pre-S2 binding to hepatocyte by 40% and 70%, respectively. Added extraneous polymerized albumin and the antibody against it did not interfere with virus attachment to HepG2 cells.

Cross Reactions↗

A light & electron microscopic study on apoptotic bodies in acute aflatoxin B1 induced injury in the rat liver.

A subnecrogenic dose of aflatoxin B1 (AFB1) was injected intraperitoneally into 8-10 wk old male rats of the Holzman strain. Cytoplasmic and sinusoidal eosinophilic bodies were seen in the liver which appeared at 30 h and reached a maximum at 48 h. Electron microscopically some of the cytoplasmic structures were seen to be of mitochondrial origin whereas others resembled apoptotic bodies. The sinusoidal bodies were similar to apoptotic bodies.

Aflatoxin B1↗

Enterically transmitted non-A, non-B hepatitis: recovery of virus-like particles from an epidemic in south Delhi and transmission studies in rhesus monkeys.

An epidemic of viral hepatitis, serologically characterized as due to non-A, non-B hepatitis, occurred in a village of South Delhi, India, in December, 1986, through January, 1987. Water contaminated with fecal matter was the apparent source of infection. Disease-associated virus-like particles were detected by immune electron microscopy in the feces of three patients within 5 days of illness. The virus-like particles were agglutinated by autologous acute-phase serum but not by convalescent serum. Rhesus monkeys inoculated with particle-containing fecal suspensions developed biochemical and morphologic features of acute, self-limited hepatitis. The findings in the present study and in earlier investigations of sporadic non-A, non-B hepatitis suggest that (i) the epidemic form and a proportion of sporadic cases of this infection in India may be related, both being enterically transmitted and associated with infection by a 27- to 32-nm virus-like particle, (ii) antibody responses to this virus occur early in disease and are transient and (iii) the rhesus monkey may prove to be a suitable model for studies of epidemic non-A, non-B hepatitis.

Adolescent↗

Enhancement of aflatoxin B1-induced hepatocarcinogenesis in rats by partial hepatectomy.

The effect of enhanced cell replication induced by partial hepatectomy (PH) in aflatoxin B1 (AFB1)-induced hepatocarcinogenesis has been studied in rats of the inbred As2 strain. Animals were given 0.25 mg/kg body weight of AFB1 as a single intraperitoneal dose 24 h after PH. Non-hepatectomized animals given the same dose of AFB1 served as controls. Neoplastic nodules and hepatocellular carcinoma (HCC) were detected respectively in 100% and 90% of hepatectomized animals sacrificed between 55 and 65 weeks after AFB1 administration. None of the ten non-hepatectomized rats sacrificed at this time interval showed HCC or neoplastic nodules. On histochemical staining the tumour population was found to be heterogeneous. Thus PH resulted in enhancement of AFB1-induced hepatocarcinogenesis in rats of the AS2 strain.

Aflatoxin B1↗