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Biomedical subjects

A Cataldi

Publications and source records attributed to A Cataldi.

At least 73 records · Page 4Linked to original sources
PubMed1997

Phospholipase C gamma 1 overexpression and activation induced by interferon beta in human T lymphocytes: an ISGF3-independent response.

Interferons exert their antiviral, antiproliferative and immunoregulatory activities by stimulating the expression of several genes. Such genes disclose a common element within their promoters, defined Interferon Stimulated Response Element (ISRE), which binds a nuclear factor(s) translocated from the cytoplasm to the nucleus (ISGF3) after the binding of interferon (IFN) to the specific receptor. Here we report the induction of the synthesis and of the hydrolytic activity of phospholipase C gamma 1 (PLC gamma 1) in human T lymphocytes by IFN-beta. The increased level of PLC gamma 1 becomes evident after 90 min of IFN-beta treatment and is still detectable after 24 h. Neither the PLC gamma 1 overexpression induced by IFN nor the increased hydrolytic activity of the enzyme appear to be affected by pretreatment of the cells with the protein tyrosine kinase inhibitor genistein, which is known to prevent the association of ISGF3 components. These results suggest that in human T lymphocytes IFN-beta can activate other transcription factor(s) distinct from ISGF3 to regulate PLC gamma 1 expression. In addition, the ability of this enzyme to hydrolyse PIP2, also in the presence of genistein, implies the possibility that this enzyme can exert its hydrolytic activity independently of protein tyrosine kinase activation.

Blotting, Western↗
PubMed1997

An mpb-64 flanking sequence specific for Mycobacterium bovis.

A clone carrying a plasmid with the mpb-64 gene and 3' flanking sequences (plasmid pMBA122) was detected during the screening of a Mycobacterium bovis genomic library with sera from infected cattle. When the pMBA122 insert was used as a probe in Southern blots against PvuII-digested mycobacterial DNA, it distinguished the different M. tuberculosis complex species. This probe hybridized with a 7-kb band in M. tuberculosis, a 5-kb band in M. bovis and a 3-kb band in M. tuberculosis complex strains from wild seals. Smal genomic digestions enabled us to locate this polymorphic region downstream of the mpb-64 gene. In order to clone this particular region, we designed a pair of PCR primers. Unexpectedly, these primers amplified only M. bovis DNA; no amplification was seen in M. tuberculosis DNA. When the annealing temperature was lowered from 70 to 55 degrees C, an amplification product of the same size was obtained with M. tuberculosis. This product was cloned and sequenced, and showed partial homology to the M. bovis amplified fragment. Therefore, this region comprises M. bovis sequences with a lower homology with M. tuberculosis than other compared sequences. This suggests that a more precise differentiation method at the species level for the M. tuberculosis complex could be achieved using PCR directed to this region.

Animals↗
PubMed1997

A novel 27 kDa lipoprotein antigen from Mycobacterium bovis.

A novel Mycobacterium bovis antigen was identified from an expression library using sera from naturally infected cattle. The Escherichia coli recombinant clone expressed a 27 kDa protein, named P27. A rabbit serum against the recombinant antigen recognized a protein of 27 kDa in cellular extracts from M. bovis and M. tuberculosis. No protein was recognized in the culture supernatant. Sequence analysis indicated that P27 has a molecular mass of 24 kDa, showing a characteristic signal sequence for lipoprotein modification (a signal peptidase type II site). The gene is identical to a gene identified in the M. tuberculosis genome sequencing project. Cellular fractionation experiments suggested that P27 is an integral membrane protein. The antigen was recognized by individual sera and peripheral blood mononuclear cells (PBMC) from diseased cattle. PCR experiments with specific primers directed to the P27 structural gene indicated that it is only present in the M. tuberculosis species complex. In conclusion, a novel immunogenic lipoprotein in M. bovis/M. tuberculosis has been identified. The results presented here and elsewhere suggest that mycobacterial lipoproteins should be considered in the design of new recombinant vaccines and diagnostic methods.

Amino Acid Sequence↗
PubMed1996

Comparison of different genetic markers for molecular epidemiology of bovine tuberculosis.

In the present study three different genetic markers were used in an RFLP study to differentiate Mycobacterium bovis (M. bovis) isolated in Argentina. The markers were: the insertion sequence IS6110, the direct repeat (DR) sequence flanking IS6110, and a polymorphic GC-rich repetitive sequence (PGRS), called pMBA2. Two restriction enzymes were used, PvuII for IS6110 and DR (DR/PvuII) and AluI for DR (DR/AluI) and pMBA2. DNA from 85 of M. bovis isolates was digested with PvuII and hybridized with IS6110 and Dr. IS6110 was not useful to differentiate M. bovis because most of the isolates contain a single monomorphic copy. The use of DR allowed a limited degree of differentiation. DNA from 44 of these isolates was also digested with AluI and hybridized with DR and pMBA2. In this condition these probes differentiated the isolates in many different RFLP types. By combining the patterns generated with DR/AluI and pMBA2 it was possible to increase the differentiation up to obtain 30 different RFLP types and 54% of the isolates were differentiated because they showed a unique pattern. Six isolates of M. bovis involved in two different outbreaks of bovine tuberculosis were correctly identified. Thus, DR and pMBA2 could be, at the moment, the probes of choice for comparisons of M. bovis isolates in different regions and for epidemiological surveillance of bovine tuberculosis.

Animals↗
PubMed1996

Effect of the 5-HT3 receptor antagonist ondansetron on plasma AVP secretion: a study in cancer patients.

The aim of the present study was to investigate the effects of a serotonin subtype 3 receptor antagonist, ondansetron, on arginine vasopressin secretion in humans. Plasma vasopressin concentrations were determined in 24 breast cancer patients undergoing adjuvant chemotherapy, before and after ondansetron intravenous (i.v.) administration. Ondansetron (8 mg i.v. at time 0 and 8 mg po at time 240 min) was administered alone in 12 patients and afterwards in combination with chemotherapy in all patients. No changes in hormone levels were found after ondansetron alone and in 17 patients who did not claim nausea and/or emesis after chemotherapy. In seven patients who experienced nausea and /or emesis, vasopressin levels significantly (P < 0.01) increased (from 6.3 +/- 0.9 ng/L in basal conditions to 15.1 +/- 3.3 ng/L at 10 h; P < 0.05 vs baseline). The results suggest the possibility that in humans, serotoninergic mechanisms, which modulate vasopressin secretion, may involve the activation of the serotonin receptors recognised by ondansetron.

Aged↗
PubMed1996

Morphological patterns and DNA polymerase regulation in apoptotic HL60 cells.

An ultrastructural and functional study was performed during methotrexate (MTX) induced apoptosis on HL-60 leukemia cells. The fine preservation of plasma membrane architecture and organellar components was present until latest apoptotic stages, despite strong nuclear changes. This observation suggests the presence of a residual cell metabolic activity, which is here investigated. DNA agarose gel electrophoresis demonstrated its fragmentation, which was also confirmed in situ by nick translation confocal analysis. Nuclear purification was subsequently performed to investigate DNA polymerase activities. DNA polymerase alpha activity appeared strongly reduced from the early phases of methotrexate treatment, while the rate of DNA polymerase beta synthesis was found to increase progressively along with the time of drug treatment. Low levels of DNA polymerase gamma activity revealed both in control and in treated cells, suggesting the irrelevant involvement of this enzyme in the DNA metabolism of this model. DNA polymerase beta appears thus to be the enzyme preminently correlated to cell attempts to repair the methotrexate-induced DNA damage.

Apoptosis↗
PubMed1996

Tuberculosis in sea lions and fur seals from the south-western Atlantic coast.

Diverse pathological conditions causing the strandings and/or deaths of several species of sea lions and seals on the northern coast of the province of Buenos Aires are being studied. Tuberculosis was diagnosed in six cases of strandings, involving two otariid seal species (one Otaria flavescens and five Arctocephalus australis), between March 1989 and December 1992. Necropsies were performed on all six cases. Granulomatous lesions were observed in the prescapular and hepatic lymph nodes. Lesions were also seen in the lungs, pleura, liver, spleen and peritoneum. Bacteriological isolation was attempted from all the samples. The isolates were identified as belonging to the Mycobacterium tuberculosis complex. Some showed characteristics consistent with M. bovis, whereas others demonstrated properties of M. tuberculosis. Genomic deoxyribonucleic acid (DNA) from these strains was analysed by restriction fragment length polymorphism (RFLP), using IS6110, a genetic marker found only in the Mycobacterium tuberculosis complex. Using the IS6110 probe, similar fingerprints were obtained, suggesting a common source of infection. However, the pattern of DNA differed from DNA patterns of M. bovis isolated from humans and cattle in Argentina, which generally contain a unique 1.9 kbp band. These results suggest that mycobacteria isolated from wild seals form a different grouping inside the M. tuberculosis complex. This is the first time that tuberculosis has been detected in wild seals from the south-western Atlantic coast.

Animals↗
PubMed1996

Experimental study on protein transmission through the human muscle fascia: preliminary results and application theory in lymphedema.

The presence of proteins (albumin and globulins) in lymphedematous tissue not only gives rise to colloidosmotic pressure but also produces an electrostatic charge endowing the proteins with individual features and different migration rates. The working hypothesis of the experimental study is to transfer lymph proteins from the upper fascia accumulation area to a subfascial drainage area by subjecting them to an adequate difference in potential. A double chamber, variable volume system with separation wall able to contain a 1 cm square of muscle fascia, was designed and built; the aim of the apparatus was to reproduce the subcutaneus zone separated by the fascia interposition, from the muscle-vascular zone. At the system was applied a variable electric field in six different experiments: 4 using porous synthetic membranes and 2 using human muscle fascia.

Biological Transport↗
PubMed1996

Multidrug resistant tuberculosis outbreak in Buenos Aires. DNA fingerprinting analysis of isolates.

In order to determine the possible relationship among HIV patients coinfected with multidrug resistant tuberculosis strains who had been receiving clinical assistance in our Hospital, clinical and epidemiological information from 28 patients was collected. DNA fingerprinting by restriction fragment length polymorphism (RFLP) pattern was performed on the mycobacterial isolates from these patients, using the restriction enzyme Pvull and IS 6110 as genetic marker. A unique RFLP pattern was found in 10 isolates from 10 different patients who had a disease caused by a single strain. Our findings confirm RFLP as a reliable and useful tool to analyze TB transmission.

Acquired Immunodeficiency Syndrome↗
PubMed1996

[Contribution of a combination of alpha and beta benzopyrones, flavonoids and natural terpenes in the treatment of lymphedema of the lower limbs at the 2d stage of the surgical classification].

HYPOTHESIS: To create a phlebolymphologic therapy in order: to activate venous system; to activate lymphatic system; to activate macrophagic system; to reduce the proteic lymphatic load. EXPERIMENTAL: A study was performed on the use of an ideal phlebolymphological association (Tonka Beans, Gingko Biloba, Melilotus Officinalis) as a practical standpoint in the treatment of lymphedema of lower limbs in order to create an efficacious dose of Coumarin, Benzopyrones and Ginkolidi. CLINICAL: We investigated a population of 76 patients treated in an open-label study for six-eight months with a dosage of Coumarin 60 mg/daily + Gingko Biloba 40 mg/daily + Melilotus 40 mg/daily. CONCLUSION: This trilogy induced a very significant improvement in lymphedema (centimeter-aspect) both in functional symptoms (pain heaviness in affected limbs) and physical signs (edema, episodes of infection). Tolerance of long term treatment was good and the improvement was observed from the third month of treatment.

Adolescent↗
PubMed1995

Tiazofurin induces a down-modulation of ICAM-1 expression on K562 target cells impairing NK adhesion and killing.

Tiazofurin treatment of K562 leukemia cells in vitro depletes the metabolites of the guanylate biosynthetic pathway, inducing erythroid differentiation, that, in turn, alters the phenotypic profile. As a consequence, K562 cells possibly modify their interaction with immune cells. Here we describe the binding and killing activity of peripheral blood NK cells against differentiating K562 cells and the correlation between their altered binding capacity and ICAM-1 expression levels in differentiating K562 cells. We found that decreased percentages of NK (and T) cells were bound to differentiating K562 cells generating a decreased cytotoxic activity. This corresponded to decreased expression of ICAM-1, as detected by FACS analysis and Western blot. Erythroid differentiation, binding and killing reduction, and ICAM-1 down-modulation were completely abrogated by guanosine treatment. Tiazofurin causes a decrease in lymphocyte recognition and binding to K562 target cells. This can be ascribed to the down-modulation of ICAM-1 expression on target cells, which, therefore, can escape killing, acquiring a selective survival advantage.

Antigens, CD↗
PubMed1995

Genetic characterization of mycobacteria from South American wild seals.

Tuberculosis has been recently diagnosed in four wild seals found stranded in the Atlantic coast of Argentina. By bacteriological studies and IS6110 hybridization, these isolates were characterized as belonging to the Mycobacterium tuberculosis complex. A genetic characterization using RFLP (Restriction fragment length polymorphism) and a species-specific probe of M. tuberculosis, called mtp40, showed hybridization with this probe on a single band. A similar band was also found in M. tuberculosis H37Rv. This showed a relationship between M. tuberculosis and the wild seal isolates. However these would also seem to belong to a different genetic group in the M. tuberculosis complex, since they do not grow on glycerol-egg containing medium (Lowenstein-Jensen) as typical M. tuberculosis strains usually do. Repeated sequences pMBA2, pTNB12, DR and IS6110 were used as probes to evaluate the epidemiological relationships between the 4 cases of tuberculosis. A low degree of polymorphism was observed, that suggested that these isolates were epidemiologically related.

Animals↗
PubMed1995

Plasma beta-endorphin levels and glucose tolerance in patients with chronic renal failure.

In order to examine the role of endogenous opioid peptides on glucose metabolism in uraemic patients, plasma concentrations of beta-endorphin, glucose, insulin and C-peptide were determined before and during an oral glucose tolerance test (OGTT) in nine non-dialysed patients with chronic renal failure (CRF). The results are compared with those obtained in a group of age-matched normal subjects. In CRF patients, plasma beta-endorphin fasting values (16.0 +/- 1.9 pmol/l) were significantly higher than those of the controls (6.6 +/- 0.6 pmol/l) and significantly correlated with the degree of renal function impairment. After glucose load, plasma beta-endorphin in CRF patients tended to decline, whereas in normal subjects increased. The fasting and the mean OGTT plasma beta-endorphin values negatively correlated with insulin initial response to glucose, insulin and C-peptide mean OGTT values, but not with glucose OGTT mean values. Data indicate that chronic uraemia induces a significant increase in circulating plasma beta-endorphin levels, with a loss of opioid system responsiveness to glucose. The possibility that this hyper-endorphinism may have a biological importance at least as a contributory factor of impaired glucose tolerance in uraemia may be suggested.

Adult↗
PubMed1995

Inositides in nuclei of Friend cells: changes of polyphosphoinositide and diacylglycerol levels accompany cell differentiation.

Friend erythroleukemia cells were labelled with high levels of [3H]myo-inositol and the radioactivity in PI, PIP and PIP2, extracted from isolated nuclei, was measured. A parallel analysis employing a picomole sensitive assay for both PIP and DAG has been carried out. The results indicate that the differentiation process is characterised by an accumulation of nuclear PIP and PIP2 and by a decrease of DAG mass. We suggest that as differentiation proceeds toward erythrocytes in Friend cells, this is accompanied by a reduction in the amount of these messengers in the nucleus.

Animals↗
PubMed1995

Phosphoinositidase C beta 1 isoform expression is modulated by interferon alpha in Burkitt lymphoma cells.

The expression of phosphoinositidase C (PIC) at nuclear and cytoplasmic level has been revealed in control and interferon treated Burkitt lymphoma cells by means of western blotting and immunocytochemical analysis employing specific monoclonal antibodies against beta 1, gamma 1 and delta 1 isozymes. Results have indicated that PIC isoform beta 1, mainly detectable in the nucleus, undergoes transient modifications early after interferon treatment. PIC delta 1 has been found only at cytoplasmic level, apparently insensitive to interferon treatment, while PIC gamma 1 was scarcely or not detected either in the cytoplasmic or in the nuclear compartment. These results suggest that interferon may exert its antiproliferative effect activating at least two distinct pathways of signal transduction, at cytoplasmic and nuclear level, involving inositol lipid cycle mainly in the nucleus by modulation of PIC beta 1 expression.

Burkitt Lymphoma↗
PubMed1995

Interferon beta mediated intracellular signalling traffic in human lymphocytes.

The early molecular mechanisms activated by the treatment of human lymphocytes with human interferon beta have been studied. These identify an early increase with respect to control, in diacylglycerol (DG) levels as response to interferon treatment. Such a DG production was derived from the rapid and sequential activation of phosphoinositide specific phospholipase C and phospholipase D pathway. This suggests that a synergistic involvement of phosphatidylinositol-bis-phosphate (PIP2) hydrolysis and phosphatidylcholine (PC) breakdown provide early molecular events upon the interaction between interferon beta and its cell surface receptors. This finally leads to the slowing down of cell growth.

Diglycerides↗
PubMed1995

Cloning of a novel polymorphic GC-rich repetitive DNA from Mycobacterium bovis.

A repetitive DNA from a wild-type Mycobacterium bovis isolate was cloned and characterized. The repeated segment was present in M. bovis and M. tuberculosis but was absent from the six other mycobacteria tested. Sequence analysis demonstrated that this repetitive element belonged to the polymorphic GC-rich repeat sequence type, a family of interspersed repeated DNA. This fragment, when used as a probe in restriction fragment length polymorphism analyses, was able to detect polymorphism in M. bovis genotypes that went undetected when the established IS6110 was used as a probe. This repetitive element should be useful in epidemiological studies of bovine tuberculosis.

Base Sequence↗