PubMed Health⌕ Search

Biomedical subjects

A Turano

Publications and source records attributed to A Turano.

At least 55 records · Page 3Linked to original sources
PubMed1995

Purification of LamB proteins using continuous elution electrophoresis: a comparison with immunoaffinity chromatography.

LamB is a membrane protein that allows the exposition of a foreign peptide on the surface of a recombinant E. coli cells. An immunopurified hybrid LamB protein has been used to elicit high-titre antibodies to a foreign epitope. Looking for a simpler purification procedure we have compared the traditional approach, which includes affinity chromatography, to continuous elution electrophoresis, in the purification of two different hybrid LamB proteins as foreign epitopes. The results obtained showed that both methods yielded the same purification, although the electrophoretic procedure had a higher yield. Continuous-elution electrophoresis could be a useful tool for the purification of membrane proteins.

Antibodies, Monoclonal↗
PubMed1995

Monoclonal antibodies against antigens exposed on the surface of vegetative forms and spores of Myxococcus virescens.

Twelve monoclonal antibodies (mAbs) directed against cell-surface antigens of Myxococcus virescens cells were developed and partially characterized. All of them recognized multiple, diffuse proteic bands in Western blot and four were also reactive to living bacteria, as assessed by flow cytometry. The four latter mAbs recognized antigens common to a number of vegetative forms and spores. The selective expression of proteins recognized by mAbs on the microorganisms and the possible applications of mAbs to the study of myxobacterial cell interaction are discussed.

Antibodies, Bacterial↗
PubMed1994

Persistence of human T cell lymphotropic virus type 1 (HTLV-1) sequences in peripheral blood mononuclear cells from patients with mycosis fungoides.

Mycosis fungoides (MF) is a rare form of cutaneous T cell lymphoma suspected of having a viral etiology. As in adult T cell leukemia, the virus involved may be human T lymphotropic virus type 1 (HTLV-1). We cultured the peripheral blood mononuclear cells (PBMC) of 29 patients with MF HTLV-1 seronegative by enzyme-linked immunosorbent assay and Western blot. The presence of reverse transcriptase (RT) and p24 antigen was investigated in the concentrate supernatant of the culture. The DNA of all studied patients was submitted to polymerase chain reaction and Southern blot analysis using primers and probes recognizing the tax region of HTLV-1/2 and the pol region of HTLV-1. 10 of 29 patients were found positive to HTLV-1, whereas they were always negative to RT and p24. The same results were confirmed in double blind after 6 mo. Our findings suggest HTLV-1 may be involved in the etiology of MF, at least in certain cases.

Adult↗
PubMed1994

Diagnosis of bronchopulmonary infections by quantification of microflora.

The quantification of bacteria and fungi in sputum or bronchoaspirate is of clinical value for the diagnosis of respiratory tract infections. We have developed an easy method to count the micro-organisms in patients with respiratory tract infections. This consists of the quantification of micro-organisms by subsequent streakings of a calibrated loop on agar. The correlation between microbiological quantitative data and the clinical status of patients with lower respiratory tract infections is discussed. The data seem to indicate that certain bacteria present in sputum or bronchoaspirate above a certain concentration may be responsible for lower respiratory tract infections. In patients with immunological disorders or chronic pathologies even lower concentrations of micro-organisms in bronchial secretions probably are enough to cause infections. The advantage of this counting method of the microbic species from the respiratory tract consists of their quantification: thus we can attribute an etiological role to a high concentration of the germs, while micro-organisms at low concentrations are probably contaminants. By this method isolated colonies are obtained after 12-18 hours. The bacterial quantification, by respiratory samples examination of the same patient in the following days, allows us to evaluate the efficacy of antibacterial therapy, producing a reduction of bacterial concentration.

Acquired Immunodeficiency Syndrome↗
PubMed1994

A multicentre study: Staphylococcus and Enterococcus susceptibility to antibiotics.

A multicentre study to evaluate the susceptibility of Gram-positive cocci isolated from clinical samples, was performed by six centres working in different areas of Italy. We examined 4,544 strains of Staphylococcus aureus, 4,381 strains of coagulase-negative staphylococci and 2,478 strains of enterococci. The following antibiotics were tested: penicillin G, ampicillin, amoxicillin, piperacillin, imipenem, oxacillin, ofloxacin, pefloxacin, ciprofloxacin, gentamicin, tobramycin, amikacin, netilmicin, rifampicin, clindamycin, tetracycline, cotrimoxazole, erythromycin, chloramphenicol, vancomycin and teicoplanin. Oxacillin-susceptible staphylococci confirmed their susceptibility to many other antimicrobial agents while oxacillin-resistant strains confirmed their multiple and frequent resistance to antibiotics. Resistance to oxacillin, cotrimoxazole and chloramphenicol was more frequent in coagulase-negative staphylococci than in Staphylococcus aureus. Aminoglycosides, rifampicin and quinolones were more active against coagulase-negative staphylococci than against Staphylococcus aureus. Enterococci were susceptible to penicillins and imipenem, and moderately susceptible to ciprofloxacin. Susceptibility of 70-79% was observed with high levels of aminoglycosides. Excellent results against staphylococci and enterococci were observed with vancomycin and teicoplanin.

Anti-Bacterial Agents↗
PubMed1994

In vitro antibacterial activity of the new quinolone Bay y3118 against clinical isolates.

The in vitro antibacterial activity of the new fluoroquinolone Bay y3118 against 609 clinical isolates was evaluated. Bay y3118 exhibited activity against a broad spectrum of organisms, including Gram-negative bacilli, Gram-positive cocci, mycobacteria. The activity of Bay y3118 was often superior to that of other quinolones. Against Gram-negative bacilli its activity was similar to that of ceftriaxone, cefotaxime, ceftazidime and imipenem except for Serratia marcescens, Klebsiella pneumoniae, Enterobacter spp. and Xanthomonas maltophilia, where its activity was superior. Gentamicin and piperacillin sometimes were less active. Bay y3118 was active against a large number of Gram-positive cocci. The fluoroquinolones tested were active against all the strains of Mycobacterium tuberculosis, but only Bay y3118 was effective against Mycobacterium avium.

Anti-Bacterial Agents↗
PubMed1994

Development of a flow cytometric assay for the detection and measurement of neutralizing antibodies against human immunodeficiency virus.

Cells infected with human immunodeficiency virus type 1 (HIV-1) develop viral antigens which can be detected by immunofluorescence. We developed a flow cytometric immunofluorescence assay (FIFA) which provides a quantitative analysis of HIV-1 p24 using a monoclonal antibody (mAb) as a specific reagent. The reduction of HIV p24 antigen expression in viral infected cells was then used to determine HIV neutralizing antibody titers in human sera. Results obtained by FIFA for detecting neutralizing antibodies against HIV-1, when compared with results obtained by indirect immunofluorescence assay (IFA), showed an overall index of agreement of 94.1%. The correlation between the neutralizing antibody titers obtained with each method was found to be highly significant (ro = 0.8; p < 0.01). The simple methodology and the adaptability of this FIFA to various assay conditions make it suitable for routine purposes and for assessing the efficacy of vaccination strategies.

Antibodies, Monoclonal↗
PubMed1993

Flow cytometric indirect immunofluorescence assay with high sensitivity and specificity for the detection of antibodies to HSV-1 and HSV-2.

Cells infected with HSV-1 or HSV-2 develop viral antigens which can be detected by immunofluorescence. We developed a flow cytometric indirect immunofluorescence assay to detect and quantitate antibodies to HSV-1 and HSV-2 in human sera. Results obtained by flow cytometry for detecting antibodies against HSV-1, when compared with results obtained by ELISA, showed an index of overall agreement of 100%. The correlation between the antibody titers obtained with each method was found to be highly significant. An index of overall agreement equal to 94.1% was observed between results obtained by flow cytometry and by immunofluorescence as concerns the discrimination of HSV-2 positive from negative samples. However, the correlation between antibody titers was found to be not statistically significant. The flow cytometric assay proved to be type-specific.

Animals↗
PubMed1993

Expression of activation markers on peripheral-blood lymphocytes following oral administration of Bacillus subtilis spores.

This study was undertaken to assess the capability of Bacillus subtilis spores to modify the peripheral-blood lymphocyte (PBL) subsets or determine the de novo expression of activation markers. The data we obtained show that spores of B. subtilis are able to increase the expression of certain cell activation markers and that such activation is dose-dependent. In fact, doses of 2 x 10(9) spores did not give rise to changes in any of the parameters evaluated, while doses of 6 x 10(9) increased the HLA-DR antigen expression on T-lymphocytes. At the highest dosage used (12 x 10(9), B. subtilis spores caused the appearance of cells bearing the CD25 and CD71 activation markers. Therefore, such cell activation markers may prove useful for monitoring the activity of B. subtilis spores, and possibly of other immunomodulating agents, in the course of clinical research.

Administration, Oral↗
PubMed1993

High-titre antibodies to a foreign epitope elicited by affinity-purified hybrid LamB proteins.

A monoclonal antibody to the LamB protein, named LBS-1, was developed and characterized. It was then covalently bound to Sepharose and used to purify hybrid LamB proteins from Escherichia coli crude extracts. A peptide of the interferon-gamma (IFN-gamma) NH2-terminal region, inserted within the LamB protein, was used as a model to assess immune response in mice injected with sonicated E. coli extract or with affinity-purified hybrid LamB protein. None of the mice immunized with the whole bacterial extract produced antibodies to IFN-gamma. On the other hand, all the mice immunized with the purified protein developed high-titre anti-IFN-gamma antibodies. These results might be due to the presence of bacterial components capable of masking the LamB protein to the immune system. The use of affinity-purified LamB proteins may constitute in some instances a more effective way of generating an immune response against foreign epitopes as opposed to whole bacterial antigens.

Amino Acid Sequence↗
PubMed1993

The role of human autoantibodies against gamma-interferon.

Natural antibodies to gamma-interferon (IFN-gamma) were detected in the serum of viral infected patients and also in the serum of healthy subjects at a lower titre. The increased titres of antibodies to IFN-gamma in sera from viral infected patients, and their decrease with clinical resolution, indicates that these antibodies are related to the viral infection and probably reflect IFN-gamma production as a result of antigenic stimulation in vivo. These antibodies were affinity-purified and studied for their capability to interfere with the multiple activities of IFN-gamma in vitro. Data obtained show that human anti-IFN-gamma antibodies interfere with the immunomodulating activity but not with the antiviral and antiproliferative activity of lymphokine. Autoantibodies to IFN-gamma may have a role in the immunoregulatory process serving to limit the intensity and or duration of the immune response. Being able to interfere with the immunomodulating activities of IFN-gamma, these antibodies might open up new therapeutic possibilities for those diseases which carry evidence of activated cell-mediated immunity.

Acquired Immunodeficiency Syndrome↗
PubMed1993

Rapid methods for microbial diagnosis of mycobacterial infections.

The Authors describe their experience in rapid diagnosis of mycobacterial infections using a combination of a radiometric blood culture (Bactec 13 A) and a nucleic acid hybridization system (Gen probe, Accuprobe) to detect and identify Mycobacteria. They found out that a high number of septicaemias in HIV positive patients are due to Mycobacterium avium, while in HIV negative subjects Mycobacterium tuberculosis is the most frequent mycobacterium.

HIV Seropositivity↗
PubMed1993

HTLV-1 sequences in lymphocytes of a patient with non-tropical spastic tetraparesis and her asymptomatic husband.

We describe a 65-year-old woman born and living in Northern Italy affected by non tropical spastic tetraparesis and her asymptomatic husband presenting HTLV-1 sequences in their lymphocytes detected by polymerase chain reaction (PCR). We discuss the significance of the case and the mechanism involved in HTLV-1 infection and the relationship with neurological disorders, stressing that this case is the first reported in Italy.

Blotting, Southern↗
PubMed1992

Natural human antibodies to gamma interferon interfere with the immunomodulating activity of the lymphokine.

Natural antibodies to gamma interferon (IFN-gamma) were found in patients suffering from different viral diseases and, at a lower titer, in healthy individuals. Such antibodies were affinity-purified and studied for their capability to interfere in vitro with the antiviral and immunomodulating activity of IFN-gamma. Data obtained show that these human anti-IFN-gamma antibodies have no inhibitory effect on the antiviral activity of IFN-gamma. On the contrary, they are able to inhibit the expression of Fc receptor sites and HLA-DR antigens induced by IFN-gamma on the U-937 cells, a human monocytoid/macrophage-derived cell line. These antibodies can also interfere in a mixed lymphocyte culture (MLC) with the proliferation of lymphocytes and the generation of cytotoxic lymphocytes. However, they showed only a moderate inhibitory effect on the cytotoxicity generated in MLC to K-562 cells. Human antibodies capable of interfering with the immunomodulating activities of IFN-gamma might open up a new field in clinical therapy for those diseases that carry evidence of activated cell-mediated immunity.

Animals↗
PubMed1992

New clinical data on the prophylaxis of infections in abdominal, gynecologic, and urologic surgery. Multicenter Study Group.

Two dose schedules of the antibiotic cefotaxime were compared in a prospective, randomized 226-center study of 3,670 patients undergoing abdominal, gynecologic, and urologic surgery. Schedule A consisted of a single preoperative dose and schedule B consisted of one preoperative dose followed by two postoperative doses. There was no significant difference in the frequency of wound infection or bacteriuria between the two schedules. Schedule B was associated with a significantly higher incidence of postoperative pyrexia, further antibiotic therapy, local side effects, and extended hospital stay. One dose probably has less impact on the intestinal flora. Therefore, single-dose cefotaxime is as effective and less costly when compared with multiple-dose cefotaxime for common surgical procedures lasting less than 3 hours.

Abdomen↗
PubMed1992

Inhibition of the biological activity of human interferon-gamma by antipeptide antibodies.

To study the domain(s) responsible for the different biological activities of human interferon-gamma (HuIFN-gamma), rabbits were immunized with peptides corresponding to the five most hydrophilic amino acid sequences of the lymphokine. The resulting antisera were able to recognize both the immunizing peptide and the native protein. Antibodies to the carboxy-terminal region (amino acids 125-137) of HuIFN-gamma were able to interfere with the immunomodulating, antiviral, and antiproliferative activities of the lymphokine. Inhibition of the antiproliferative and antiviral activity was also observed using antibodies raised against the amino-terminal region (amino acids 4-16) and amino acids 80-95, respectively, but to a lesser extent than that observed with antibodies to the carboxyl terminus. The capability of these antibody preparations to partially interfere with only one of the lymphokine's biological activities might be explained by a mechanism of steric hindrance. The use of polyclonal antibodies allowed us to limit the presence of epitopes responsible for recombinant (r)HuIFN-gamma biological activities to the carboxy-terminal region.

Antibodies↗