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Biomedical subjects

M Musiani

Publications and source records attributed to M Musiani.

At least 91 records · Page 5Linked to original sources
PubMed1990

In situ detection of cytomegalovirus DNA in biopsies of AIDS patients using a hybrido-immunocytochemical assay.

An in situ hybrido-immunocytochemical assay, with a digoxigenin-labelled probe, was used to show the presence of cytomegalovirus DNA in both paraffin and frozen sections from tissue blocks of 5 AIDS patients. The hybridization probe was constructed by using two different DNA fragments of the repeated sequences of the CMV genome. The CMV DNA probe hybridized in situ was immunocytochemically visualized by anti-digoxigenin Fab fragments labelled with alkaline phosphatase. This hybridization procedure proved to be sensitive, specific, and provided good resolving power. Thus, it might effectively be employed in immunohistological and virological laboratories for the diagnosis of CMV infections in AIDS patients; indeed it might even be applied further in the virological context.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Detection of B19 parvovirus infections by a dot-blot hybridization assay using a digoxigenin-labelled probe.

A non-radioactive dot-blot hybridization assay for the detection of B19 parvovirus infections was developed using a digoxigenin-labelled probe both on nylon and nitrocellulose filters. A 700 bp BamHI HindIII fragment of B19 DNA was used to construct the probe. Probe labelling was carried out by incorporating deoxyuridine triphosphate labelled with digoxigenin. The dot-blot hybridization assay was visualized by an immunoenzymatic reaction using antidigoxigenin Fab fragments labelled with alkaline phosphatase. The specificity and sensitivity of digoxigenin-labelled B19 DNA probe was compared with the results obtained with 32P-labelled B19 DNA probe. Out of the 504 serum samples tested, 3 samples were positive in all the hybridization assays performed and 494 were negative, 7 serum samples gave a weak positive reaction when Dig-B19 probe was used on nitrocellulose filters. The 77 pharyngeal swabs tested were negative in all the hybridization assays performed. Our hybridization assay showed a high sensitivity and reproducibility and it appears to be a rapid, practical and reliable test for routine screening of B19 parvovirus DNA in large numbers of clinical specimens.

Adolescent↗
PubMed1990

Rapid screening for B19 parvovirus DNA in clinical specimens with a digoxigenin-labeled DNA hybridization probe.

A rapid dot blot hybridization assay for the detection of B19 parvovirus DNA in human sera was developed. Small portions of four serum samples were mixed, filtered onto a nylon membrane, and hybridized with a digoxigenin-labeled DNA probe; for each membrane, 380 serum samples could be tested. When a dot was positive by the hybridization assay, the four serum samples dotted together were separately tested to identify the sample positive for B19 DNA. A total of 10,150 serum samples submitted for viral serological and laboratory investigation with no specific requests for B19 testing were analyzed. Nine serum samples were positive for B19 DNA by dot blot hybridization assay, and the results were confirmed by electron microscopy. This method has proven to be reliable, economical in terms of time and costs, and useful for large-scale screening of clinical specimens, both for diagnostic work and for a source of antigen.

DNA Probes↗
PubMed1990

Rapid detection of cytomegalovirus DNA in urine samples with a dot blot hybridization immunoenzymatic assay.

A dot blot hybridization immunoenzymatic assay for the rapid detection of cytomegalovirus DNA in urine samples was developed by using a digoxigenin-labeled probe which was immunoenzymatically visualized by antidigoxigenin Fab fragments labeled with alkaline phosphatase. A total of 516 urine samples from different groups of subjects were analyzed, and the hybridization assay was able to yield results within 24 h. The results obtained were compared with results for detection of cytomegalovirus antigens in infected cell cultures.

Antigens, Viral↗
PubMed1990

Enhancement of HIV-1 marker detection in cell cultures treated with mild heat-shock.

Rapid exposure to supra-optimal temperature (heat-shock) and a variety of other treatments are able to induce changes in cellular translational and transcriptional activity referred to as "the heat-shock response". The effect of heat-shock was investigated in H9 lymphoblastic cells and in peripheral blood lymphocytes infected with human immunodeficiency virus type 1 (HIV-1). The results showed that a mild heat-shock performed either immediately before infection or 7 days after infection consistently increased the recovery of p24 core antigen and reverse transcriptase activity in the supernatants of experimentally infected cell cultures. On the contrary, heat-shock had no effect on the HIV-1 marker recovery from persistently infected H9/HTLV-III cell cultures.

Cell Count↗
PubMed1990

Antibody patterns against cytomegalovirus and Epstein-Barr virus in human atherosclerosis.

The immune response against Cytomegalovirus (CMV) and Epstein-Barr virus (EBV)-induced antigens has been determined in teh sera of 36 patients with angiographically assessed atherosclerosis and compared with that of 36 matched control subjects. Significantly higher titres of antibody against CMV and EBV - induced antigens and a significant increase in CMV- and EBV- reactivated infections were found in atherosclerotic patients as compared to control subjects. Moreover serological signs of concomitant EBV and CMV infections occurred more frequently in atherosclerotic patients than in controls. One atherosclerotic patient, however, completely lacked antibody to CMV while CMV- and EBV- reactivated infections were also noted in control subjects, probably connected with their old age. Our data suggest that multiple reactivations of latent viruses may represent a consequence rather than a causal factor of atherosclerosis.

Antibodies, Viral↗
PubMed1989

A hybrido-immunocytochemical assay for the in situ detection of cytomegalovirus DNA using digoxigenin-labeled probes.

A non-radioactive hybrido-immunocytochemical assay for the detection of cytomegalovirus (CMV) DNA in infected cells was developed. Two different DNA fragments belonging to the repeated sequences of CMV genome were used to construct the hybridization probe. The probe was constructed by incorporating deoxyuridine triphosphate labeled with digoxigenin. The in situ hybridized CMV DNA probe was immunocytochemically visualized by anti-digoxigenin. Fab fragments labeled with alkaline phosphatase. This procedure permitted the DNA detection, in the nuclei of infected cells fixed at 48 h after infection, of the Towne CMV reference strain and 21 different laboratory-isolated CMV strains. Our assay demonstrated a high specificity, sensitivity and reproducibility.

Cells, Cultured↗
PubMed1989

Immunological status to Epstein-Barr virus and cytomegalovirus in patients with genital condylomata.

Serological patterns against Epstein-Barr virus (EBV) and Cytomegalovirus (CMV) were determined in patients with genital condylomata (GC). The Ig G antibody values to EB-induced virus capsidic antigens (VCA), early antigens (EA) and Ig M to VCA were significantly higher in the study group than in the controls. Moreover, the concomitant presence of EBV-Ig G anti-VCA greater than or equal to 1/320, EBV-Ig G anti-EA greater than or equal to 1/20 and EBV-Ig M anti VCA greater than or equal to 1/20 was observed in 13 serum samples of genital condylomata patients, while, in only 2 serum samples of the healthy controls, the same serological pattern was present. The distribution of antibody values to CMV-induced LA, EA and IEA showed a significantly increased prevalence in the study group in comparison with the controls: the concomitant presence of antibody with a titre greater than or equal to 1/320 for CMV-LA, greater than or equal to 1/20 for CMV-EA and greater than or equal to 1/20 for CMV-IEA was observed in 15 serum samples of GC patients and in only 3 serum samples of the control group. Our results suggest that the active or recent EBV and CMV infections we observed in genital condylomata patients may be a consequence of impaired immunity in these patients, but it does not exclude a possible role of EBV and CMV in perpetuating human papilloma virus-induced cell proliferation.

Adult↗
PubMed1989

An amplified dot immunoassay for the direct quantitation of adapted and wild strains of human cytomegalovirus.

A rapid and simple quantitative dot immunoassay for a cell-adapted reference strain of cytomegalovirus (CMV) and for wild strains of CMV present in clinical urine samples was developed. The assay was performed on nitrocellulose paper dotted with several dilutions of viral pellets free of cellular debris. Viral dilutions were treated with a monoclonal antibody to the major component of the viral capsid. To amplify the reaction, a three-dimensional complex of streptavidin and biotinylated horseradish peroxidase was used as the detector system. The dot immunoassay, which does not require cell cultures, yielded results within one day. A significant correlation was found between CMV titers obtained by dot immunoassay and CMV infectious units determined by immunoalkaline phosphatase staining of CMV-late antigen positive cells.

Antibodies, Monoclonal↗
PubMed1988

Rapid quantitative assay of cytomegalovirus infectivity.

A rapid quantitative assay was developed to determine cytomegalovirus infectious units. The assay yielded results in three days. The rapid quantitation of cytomegalovirus infectivity was developed by evaluating the number of cells that expressed cytomegalovirus-late antigens at the end of a single replication cycle (62 h post-infection). Late antigens were visualized by immunoalkaline phosphatase staining using a monoclonal antibody against the major viral capsid protein (66.8 kDa). Thirty-five cytomegalovirus samples were examined. The values of the infectious units obtained by immunoalkaline phosphatase staining were correlated significantly with the values determined by the classical plaque-forming technique.

Antigens, Viral↗
PubMed1988

Immunocytochemical detection of antibodies to Epstein-Barr virus nuclear antigen by a streptavidin-biotin-complex assay.

An immunocytochemical staining for the detection of antibody to Epstein-Barr virus nuclear antigen in cell smears was developed by using a three-dimensional complex of streptavidin and biotin labeled with horseradish peroxidase as the detector system. The presence of antibody against Epstein-Barr virus nuclear antigen was revealed by a dark staining of the nuclei of Raji cells. A significant correlation was found between the titers obtained with our assay and titers obtained by anticomplement immunofluorescence on 110 serum samples. Our assay did not require an Epstein-Barr virus-negative serum source for fresh complement and gave a permanent record of the reaction that could easily be observed under an ordinary light microscope.

Antibodies, Viral↗
PubMed1988

Impairment of cytomegalovirus and host balance in elderly subjects.

The titres of IgG antibody against "late antigens", "immediate early antigens", and "early antigens" induced by cytomegalovirus (CMV) and IgM antibody against "late antigens" induced by CMV were analysed in 67 geriatric subjects by immunocytochemical techniques. The titres obtained were compared with those of an adult control population. Significantly increased titres of IgG antibody against induced antigens and a significant increase in CMV reactivated infections occurred in the elderly compared with control subjects. These findings indicate that the CMV and host balance in the elderly is disturbed, leading to activation of the CMV latent carrier state that follows primary CMV infection.

Aged↗
PubMed1988

Expression of cytoskeleton components in various Epstein-Barr virus infected cell lines.

It has been shown that viruses can induce alterations in the content and distribution of cytoskeleton structures, particularly actin microfilaments and microtubules. An immunomorphologic study of the cytoskeleton components of various EBV-infected cell lines with expression of different functions of EBV genome has been performed using antibodies to each of its three major components. Intermediate filaments and microtubules were similarly represented in all examined lymphoblastoid cell lines. The distribution of actin microfilaments, on the contrary, differed significantly from cell line to cell line. It is concluded that the morphologic expression of actin might depend on the expression of EBV genome. Furthermore, some of these cell lines might represent a useful substrate for the identification of anticytoskeleton antibodies, mainly anti-actin antibodies, in human sera.

Actins↗
PubMed1987

Dot immunoperoxidase assay using monoclonal antibody for direct detection of cytomegalovirus in urine samples.

A rapid, simple dot immunoperoxidase assay for the direct detection of cytomegalovirus in clinical urine samples was developed. The assay was performed on nitrocellulose paper dotted with urine pellets free of cellular debris. Cytomegalovirus was detected with a monoclonal antibody to the capsid antigen, and the complex was visualized by immunoperoxidase staining. Positive reactions appeared as well-defined dark blue spots. Of the 87 urine samples examined, 10 proved positive in the dot immunoperoxidase assay, and 77 proved negative. The results agreed completely with the detection of cytomegalovirus-induced antigens in cell cultures inoculated with clinical specimens.

Antibodies, Monoclonal↗
PubMed1987

Comparison of the immune response to Epstein-Barr virus and cytomegalovirus in sera and synovial fluids of patients with rheumatoid arthritis.

The immune response against two herpesviruses has been determined in sera and matched synovial fluids of patients with rheumatoid arthritis (RA) and compared with that of a healthy control population. The increased level of antibody to Epstein-Barr virus (EBV) induced antigens in patients with RA resembles the antibody pattern observed against cytomegalovirus (CMV) induced antigens, which suggests the presence of a pathological condition in patients with RA that can reactivate latent viral infections. The antibody response against EBV and CMV observed in synovial fluids excludes the local production of specific antibodies against EBV and CMV antigens.

Adult↗
PubMed1987

Comparison of various cell types as substrates for the detection of antiactin antibodies in chronic liver disease.

Human cultured fibroblasts and Raji and peripheral blood mononuclear cells have been compared as substrates for the immunofluorescent detection of antiactin microfilament antibodies in sera from chronic liver disease. Raji and mononuclear cells, which exhibit a rich microfilament network, proved to be the most sensitive substrates for this purpose. The clinical relevance of these findings in the diagnosis of "autoimmune" chronic liver disease is discussed.

Actin Cytoskeleton↗
PubMed1986

Rapid diagnosis of viral infections by an alkaline phosphatase immunocytochemical method.

The use of alkaline phosphatase immunocytochemical staining was explored for the rapid diagnosis of poliovirus, adenovirus, herpes simplex virus and cytomegalovirus infections in cell cultures. In this test, viral antigens treated with their relative antibody were incubated with alkaline phosphatase-labelled antisera. The enzyme label was developed with a naphthol salt in the presence of a diazonium salt (Fast Blue) in order to obtain a blue coloured precipitate at the site of the enzyme. It is suggested that this immunocytochemical technique is valuable in the detection of viral infections and would be an appropriate test to use when rapid diagnosis is required.

Adenoviruses, Human↗