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Biomedical subjects

C Chezzi

Publications and source records attributed to C Chezzi.

At least 19 recordsLinked to original sources

Detection of IgA and IgM antibodies to HIV-1 in neonates by radioimmune western blotting.

OBJECTIVE: To detect infection with HIV-1 by IgA and IgM response at birth in children born to HIV-1 seropositive mothers. DESIGN: Western blotting and radioimmune western blotting on stored sera from infected and uninfected babies born to HIV-1 seropositive mothers. Sera were pretreated to remove IgG. SETTING: Parma and Bologna, Italy. SUBJECTS: 12 infected and five uninfected babies born to HIV-1 seropositive mothers and three babies born to seronegative mothers. MAIN OUTCOME MEASURES: Effectiveness of western blotting and radioimmune western blotting in detecting antibodies to HIV-1 gene products. RESULTS: With conventional western blotting we found IgA class antibodies to HIV-1 proteins in serum from three out of 12 infected children; in two of these three the serum was collected at age 3 months (positive controls). Radioimmune western blotting detected both IgA and IgM antibodies in serum from all infected children tested, whereas all serum from uninfected children born to seropositive and seronegative mothers showed no such antibodies. CONCLUSION: Although the technique should be tested on more patients, radioimmune western blotting seems to be a valuable tool for serological diagnosis of congenital HIV-1 infection at birth in neonates born to seropositive mothers.

Blotting, Western

Dr. Libero Ajello.

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Centers for Disease Control and Prevention, U.S.

Anaerobic yeast killer systems.

The influence of anaerobic conditions on the expression of the killer phenomenon of several yeast isolates belonging to recognized killer systems coded by different genetic determinants (Pichia spp., Kluyveromyces lactis, Saccharomyces cerevisiae) was studied. Anaerobiosis influenced the activity of killer toxins from some individual isolates of the genera Pichia and Saccharomyces on sensitive strains of P. anomala, K. lactis and Candida albicans. However, no influence was detectable on a S. cerevisiae sensitive isolate. Thus, anaerobic conditions seem to interfere more with the metabolic process of sensitive strains than with toxin production by killer yeasts. The selection of a panel of killer yeasts, able to display their activity against reference sensitive yeast isolates under anaerobic conditions in a medium that favored the growth of anaerobes, allowed the use of the killer system to type Bacteroides fragilis isolates for epidemiological purposes.

Anaerobiosis

Susceptibility of frequent urinary pathogens to fosfomycin trometamol and eight other antibiotics: results of an Italian multicenter survey.

In order to assess the resistance profile for fosfomycin trometamol after several years of clinical use in Italy, this study has explored the susceptibility to fosfomycin and eight other antibacterial drugs of 6,021 strains isolated from 23,816 urines during 1990 in three teaching hospitals located in Genoa, Parma and Catania. Gram-negative strains, notably Escherichia coli (41.6%), were primarily involved. Amoxicillin was the least active compound with resistance in 41.4% of the isolates. Fosfomycin showed the lowest rate of resistance in both gram-negative (2.8%) and gram-positive (2.1%) pathogens. This was followed by norfloxacin with a resistance rate of 11.8% and netilmicin with 12.2%. These results indicate that fosfomycin-trometamol may continue to be used in single-dose treatment of urinary tract infections even in the absence of microbiological data since the prevalence of resistance to the drug is, at present, so low that therapeutic failure is highly improbable.

Anti-Bacterial Agents

Patterns of cytodistribution of prosomal antigens on the vimentin and cytokeratin networks of monkey kidney cells.

Prosomes were found as mRNA-associated ribonucleoprotein particles (RNP) and cofactors of untranslated (ribosome-) free mRNP. Previous data have shown the presence of prosomal networks in the cytoplasm of PtK1 and HeLa cells and their superposition onto the intermediate filaments (IF) of cytokeratin type but little if any of vimentin type. Here it is shown that in LLC-MK2 cells various prosomal antigens are present on both, vimentin and cytokeratin networks, individual prosomal antigens superposing to variable degrees onto the IF subnetworks. Some prosomal antigens in variable relative concentrations were also observed in the nuclei of these cells. We suggest the existence of prosomal subnetworks specific for each prosomal antigen superposing to a variable extent onto the IF of both types.

Animals

Rhodococcus equi cavitary pneumonia in HIV-infected patients: an unsuspected opportunistic pathogen.

Two patients seropositive for human immunodeficiency virus (HIV) and with no previous acquired immunodeficiency syndrome-defining conditions developed cavitary pneumonia and pleural disease caused by Rhodococcus equi. R. equi was isolated from these patients' sputum and lung biopsy specimens, respectively, but the microorganism was initially considered to be a contaminant (patient 1) or misidentified as a nontuberculous mycobacterium (patient 2). The R. equi infection was fatal in one patient, who died after 4 months without specific antimicrobial therapy; the second patient was unresponsive to combination therapy with various antimicrobial agents. R. equi may cause life-threatening infections in HIV-infected patients. Microbiology laboratories should be cognizant of the need to exclude R. equi as a cause of infection in highly immunosuppressed patients.

AIDS-Related Opportunistic Infections

Use of monoclonal anti-haemagglutinin antibodies for the "in vitro" selection of a sequential influenza virus antigenic variant.

A sequential antigenic variant of the A/Texas/77 (H3N2) influenza virus was obtained in vitro using a monoclonal antibody against the haemagglutinin (HA) of the antigenic variant V18 previously selected in vitro from the parental Texas virus. The sequential antigenic variant, designated DV1, the V18 antigenic variant and the parental A/Texas/77 viruses were used to evaluate the frequency of anti-haemagglutinin antibodies in human sera in single radial haemolysis assays. Twenty six of 100 children's sera, which contained antibodies to the parental A/Texas/77 virus, failed to react with the V18 antigenic variant. A higher proportion of sera (42%) failed to react with the DV1 antigenic variant with alterations in two different antigenic determinants with respect to the parental virus. The results are discussed in relation to the mechanism of antigenic drift and to the in vivo reaction of antigenic variants selected in vitro.

Antibodies, Monoclonal

Differential toxinogenesis in the genus Pichia detected by an anti-yeast killer toxin monoclonal antibody.

The differential toxinogenesis of 25 isolates belonging to species of the potential yeast killer genus Pichia that were previously classified in the genus Hansenula was comparatively demonstrated by two serologic techniques (indirect immunofluorescence and double immunodiffusion) by using a monoclonal antibody against a yeast killer toxin produced by a selected strain of Pichia anomala (UCSC 25F). The killer phenotypes of the Pichia isolates were evaluated by their ability to kill each other. The results, although of insufficient taxonomic value for a reliable separation of either species or genera, attest to the genomic heterogeneity for the killer character in the genus Pichia as well as the presumptive dual killer/sensitive identity for each single isolate.

Antibodies, Monoclonal

'Antibiobodies': antibiotic-like anti-idiotypic antibodies.

Pathogenic micro-organisms such as Candida albicans may be susceptible to the activity of antimicrobial products like yeast killer toxins due to the presence of specific cell wall receptors for these agents. Anti-idiotypic antibodies (anti-Ids) were produced that competed for these receptors with the yeast killer toxin of a strain of Pichia anomala. We report here that affinity chromatography purified anti-Ids may kill C. albicans cells in vitro which are susceptible to the activity of the yeast killer toxin, as well as P. anomala killer cells which are obviously immune to their own toxin despite possessing specific cell wall receptors which can be detected by indirect immunofluorescence with anti-Ids. We propose that these conceptually new antimicrobial immunoglobulins acting as antibiotics be called 'antibiobodies'.

Anti-Bacterial Agents

Interfaces of the yeast killer phenomenon.

A new prophylactic and therapeutic antimicrobial strategy based on a specific physiological target that is effectively used by killer yeasts in their natural ecological competition is theorized. The natural system exploited is the yeast killer phenomenon previously adopted as an epidemiological marker for intraspecific differentiation of opportunistic yeasts, hyphomycetes, and bacteria. Pathogenic microorganisms (Candida albicans) may be susceptible to the activity of yeast killer toxins due to the presence of specific cell wall receptors. On the basis of the idiotypic network, we report that antiidiotypic antibodies, produced against a monoclonal antibody bearing the receptor-like idiotype, are in vivo protecting animals immunized through idiotypic vaccination and in vitro mimicking the antimicrobial activity of yeast killer toxins, thus acting as antibiotics.

Adsorption

Defective insertion of haemagglutinin as a cause of abortivity of influenza A viruses in HeLa 229 cells.

A number of experimental data demonstrate that certain mammalian cells are unable to replicate Influenza viruses type A. In these cellular hosts the viruses can efficiently perform their biological as well as biochemical activities but the production of mature viral particles is greatly restricted. Here we report a study of abortiveness of human and avian type A Influenza viruses in HeLa 229 cells in which the final stages of maturation of viral particles seem to be affected. We show that the incorrect insertion of virus-coded haemagglutinin into the plasma membrane might be the cause of the unpermissive condition of infection exhibited by this cellular host.

Animals

Detection by immunofluorescent anti-idiotypic antibodies of yeast killer toxin cell wall receptors of Candida albicans.

Yeast killer toxin cell wall receptors of Candida albicans were observed by indirect immunofluorescence using an affinity purified rabbit anti-idiotypic antiserum. The antiserum had been raised against a monoclonal antibody neutralizing the in vitro activity of a killer toxin produced by a selected strain of Hansenula anomala UCSC 25F. This simple procedure permitted the location of killer toxin cell wall receptors in various morphological phases of the yeast cells. The use of the indirect immunofluorescence technique with anti-idiotypic antibodies may have potential value in determining the occurrence of killer toxin receptors in other microbial systems.

Antibodies, Anti-Idiotypic

The diagnosis of small bowel bacterial overgrowth. Reliability of jejunal culture and inadequacy of breath hydrogen testing.

The reliability of a single jejunal culture in the diagnosis of small bowel bacterial overgrowth has recently been questioned. Seventy-seven patients thought to have bacterial overgrowth, defined as a jejunal culture yielding at least 10(6) organisms per milliliter of aspirate, took part in the study. Bacterial overgrowth was found in 74% of the patients with predisposing conditions and in 32% of those with no clear causes of bacterial colonization. The intestinal juice of some patients was taken at two different levels of the proximal jejunum, using both the closed- and open-tube systems. Highly significant correlations (rs = 0.90, p less than 0.001) were found between the numbers of bacteria per milliliter at the 2 jejunal levels and between the numbers of bacteria per milliliter of jejunal aspirate obtained from the closed and open tubes (rs = 0.84, p less than 0.001). Compared with the jejunal culture, the gas chromatography of volatile fatty acids in jejunal aspirate and the glucose- and lactulose-hydrogen breath tests showed sensitivities of 56%, 62%, and 68% and specificities of 100%, 83%, and 44%, respectively. This work demonstrates the reliability of jejunal cultures and the inadequacy of breath hydrogen testing in the prediction of positive jejunal cultures. When results of testing for volatile fatty acids in jejunal aspirates are positive, this always indicates the presence of bacterial overgrowth; thus, this procedure would avoid the more complicated, time-consuming, and costly bacteriological analysis of jejunal samples.

Bacterial Infections

Abortive replication of influenza A viruses in HeLa 229 cells.

Several experimental data support the idea that certain mammalian cells are unable to replicate influenza viruses type A, although these viruses can efficiently penetrate the cells. This cannot be attributed to a lack of specific receptors on the cell surface, but depends upon the failure of specific step(s) to occur during viral growth. Here we report a study of abortiveness of human and avian type A influenza viruses in HeLa 229 cells. Viral polypeptide synthesis was monitored by [35S]methionine pulse labelling at several time points after infection, showing that normal amounts of virus-induced components were synthesized. Cellular fractionation of HeLa 229 cells infected by influenza viruses showed that the distribution of viral proteins into nuclear and cytoplasmic compartments was comparable to that seen in the permissive host, chick embryo fibroblasts. Viral HA glycoprotein, produced during the infectious cycle, was entirely found in the cytoplasm of infected HeLa 229 cells. The polypeptide was able to agglutinate red blood cells but did not show positive haemadsorption even at late times of infection. Therefore it seems that during the maturation of viral particles there is a failure of the haemagglutinin to perform a correct insertion into the plasma membrane of infected HeLa 229 cells.

Animals