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Biomedical subjects

A Pugliese

Publications and source records attributed to A Pugliese.

At least 55 records · Page 3Linked to original sources
PubMed1999

Effects of the human CD38 glycoprotein on the early stages of the HIV-1 replication cycle.

CD38 displays lateral association with the HIV-1 receptor CD4. This association is potentiated by the HIV-1 envelope glycoprotein gp120. The aim of this work was to evaluate the CD38 role in T cell susceptibility to HIV-1 infection. Using laboratory X4 HIV-1 strains and X4 and X4/R5 primary isolates, we found that CD38 expression was negatively correlated to cell susceptibility to infection, evaluated as percentage of infected cells, release of HIV p24 in the supernatants, and cytopathogenicity. This correlation was at first suggested by results obtained in a panel of human CD4(+) T cell lines expressing different CD38 levels (MT-4, MT-2, C8166, CEMx174, Supt-1, and H9) and then demonstrated using CD38 transfectants of MT-4 cells (the line with the lowest CD38 expression). To address whether CD38 affected viral binding, we used mouse T cells that are non-permissive for productive infection. Gene transfection in mouse SR.D10.CD4(-).F1 T cells produced four lines expressing human CD4 and/or CD38. Ability of CD4(+)CD38(+)cells to bind HIV-1 or purified recombinant gp120 was significantly lower than that of CD4(+)CD38(-) cells. These data suggest that CD38 expression inhibits lymphocyte susceptibility to HIV infection, probably by inhibiting gp120/CD4-dependent viral binding to target cells.-Savarino, A., Bottarel, F., Calosso, L., Feito, M. J., Bensi, T., Bragardo, M., Rojo, J. M., Pugliese, A., Abbate, I., Capobianchi, M. R., Dianzani, F., Malavasi, F., and Dianzani, U. Effects of the human CD38 glycoprotein on the early stages of theHIV-1 replication cycle.

ADP-ribosyl Cyclase↗
PubMed1999

Sequence analysis of the diabetes-protective human leukocyte antigen-DQB1*0602 allele in unaffected, islet cell antibody-positive first degree relatives and in rare patients with type 1 diabetes.

The human leukocyte antigen (HLA)-DQA1*0102/DQB1*0602/DRB1*1501 (DR2) haplotype confers strong protection from type 1 diabetes. Growing evidence suggests that such protection may be mostly encoded by the DQB1*0602 allele, and we reported that even first degree relatives with islet cell antibodies (ICA) have an extremely low diabetes risk if they carry DQB1*0602. Recently, novel variants of the DQB1*0602 and *0603 alleles were reported in four patients with type 1 diabetes originally typed as DQB1*0602 with conventional techniques. One inference from this observation is that DQB1*0602 may confer absolute protection and may never occur in type 1 diabetes. By this hypothesis, all patients typed as DQB1*0602 positive with conventional techniques should carry one of the above diabetes-permissive variants instead of the protective DQB1*0602. Such variants could also occur in ICA/DQB1*0602-positive relatives, with the implication that their diabetes risk could be significantly higher than previously estimated. We therefore sequenced the DQB1*0602 and DQA1*0102 alleles in all ICA/DQB1*0602-positive relatives (n = 8) previously described and in six rare patients with type 1 diabetes and DQB1*0602. We found that all relatives and patients carry the known DQB1*0602 and DQA1*0102 sequences, and none of them has the mtDNA A3243G mutation associated with late-onset diabetes in ICA-positive individuals. These findings suggest that diabetes-permissive DQB1*0602/3 variants may be very rare. Thus, although the protective effect associated with DQB1*0602 is extremely powerful, it is not absolute. Nonetheless, the development of diabetes in individuals with DQB1*0602 remains extremely unlikely, even in the presence of ICA, as confirmed by our further evaluation of ICA/DQB1*0602-positive relatives, none of whom has yet developed diabetes.

Adult↗
PubMed1998

Analysis of model for macroparasitic infection with variable aggregation and clumped infections.

A model for macroparasitic infection with variable aggregation i considered. The starting point is an immigration-and-death process for parasites within a host, as in [3]; it is assumed however that infections will normally occur with several larvae at the same time. Starting from here, a four-dimensional, where free-living larvae are explicitly considered, and a three-dimensional model are obtained with same methods used in [26]. The equilibria of these models are found, their stability is discussed, as well as some qualitative features. It has been found that the assumption of "clumped" infections may have dramatic effects on the aggregation exhibited by these models. Infections with several larvae at the same time also increases the stability of the endemic equilibria of these models, and makes the occurrence of subcritical bifurcations (and consequently multiple equilibria) slightly more likely. The results of the low-dimensional model have also been compared to numerical simulations of the infinite system that describes the immigration-and-death process. It appears that the results of the systems are, by and large, in close correspondence, except for a parameter region where the four-dimensional model exhibits unusual properties, such as the occurrence of multiple disease-free equilibria, that do not appear to be shared by the infinite system.

Animals↗
PubMed1998

Hydrops foetalis caused by hepatic haemangioma.

The authors report a moderately premature baby with Down's syndrome and hydrops, the latter probably caused by a large hepatic haemangioma which was diagnosed only after birth. At birth the baby was affected by massive right hydrothorax, ascites, hypoalbuminaemia and severe respiratory distress. With the use of modern neonatal intensive care, the baby survived. Corticosteroid treatment (prednisolone 2 mg kg(-1) d(-1) i.v. in divided doses) was associated with a very rapid resolution of the haemangioma and the baby was healthy at follow-up. Although hepatic angiomas are not uncommon in the neonatal period, the association with hydrops is a rare finding.

Adult↗
PubMed1998

[Sepsis caused by Streptococcus pneumonia in newborn infants. 2 case reports].

Streptococcus pneumoniae is responsible for 2% of all neonatal sepsis. The results of epidemiological studies suggest that newborns acquire infection by the ascending route or during the passage through the birth canal. It has been hypothesized that colonization of the maternal genital tract with S. pneumoniae might be caused by contamination of obstetric instruments with the microorganism or by sexual practices, particularly oro-genital contact. From our NICU's database, two cases of newborn sepsis due to Streptococcus pneumoniae, occurred between 1988 and 1996 have been found; the first case presented a fatal disseminated intravascular coagulation (DIC), the second a severe respiratory failure. Antibiotic treatment of women carrying S. pneumoniae in the genital tract and their infants should be strongly recommended, on the basis of the potentially serious consequences for the infants.

Age Factors↗
PubMed1997

The HIV/AIDS epidemics among drug injectors: a study of contact structure through a mathematical model.

Numerical results of a model with variable infectivity for the dynamics of HIV/AIDS (human immunodeficiency virus/acquired immune deficiency syndrome) have been compared with the data of AIDS cases among intravenous drug users in Italy, especially in the Latium region. We examined several hypotheses about the dynamics of the epidemics; for each we obtained, mainly through a least-square approach but also minimizing a different quantity, a best-fit estimate of the parameters. In the simplest model, the population is assumed to be homogeneous, and we estimate contact rate and year of start of the epidemics, obtaining a good fit up to 1989, less so after. A substantial increase in fit is obtained in assuming either a decrease of the contact rate over time or a heterogeneous population with a smaller active group. We have also compared models with different infectivity curves during the incubation period: the assumption of constant infectivity is untenable, whereas the often suggested hypothesis of a peak in infectivity shortly after infection seems to be in agreement with data.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Apoptotic DNA fragmentation, and its in vitro prevention by nicotinamide, in lymphocytes from HIV-1-seropositive patients and in HIV-1-infected MT-4 cells.

Apoptosis seems to play an important role in the decline of CD4+ T-cells in patients infected with HIV-1. Moreover, extensive interest in apoptosis comes from the observation that it correlates both with the progression and the severity of HIV-1 infection. A cross-sectional study was made to evaluate whether such correlation may also extend to the early phases of ex vivo apoptosis, after 20 h of culture. DNA fragmentation, a parameter associated with apoptosis, was evaluated with the terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling (TUNEL) technique, which preferentially labels apoptosis in comparison to necrosis. The results obtained indicate that a negative correlation exists between the proportion of lymphocytes exhibiting DNA strand breaks and the absolute number of CD4+ T-cells per microliter. DNA fragmentation was significantly higher in patients with AIDS or advanced HIV-1 infection as compared to asymptomatic patients or seronegative individuals. No significant difference was found in relation to antiretroviral therapy. Furthermore, the addition of nicotinamide to the cultures significantly reduced DNA fragmentation of both in vitro HIV-1-infected MT-4 cells and lymphocytes from six HIV-1-seropositive individuals. The results of this study confirm that DNA fragmentation, as an early marker of apoptosis, correlates with the severity of HIV-1 infection and suggest that nicotinamide may be involved in the modulation of HIV-1-related apoptosis.

Adult↗
PubMed1997

The insulin gene is transcribed in the human thymus and transcription levels correlated with allelic variation at the INS VNTR-IDDM2 susceptibility locus for type 1 diabetes.

Type 1, or insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease associated with loss of tolerance to several pancreatic islet cell molecules, including insulin, glutamic acid decarboxylase (GAD), ICA69 and the tyrosine phosphatase IA-2 (refs 1-3). Among several predisposing loci, IDDM2 maps to the insulin gene (INS) VNTR (variable number of tandem repeats) minisatellite on chromosome 11p15 (refs 4-9). Allelic variation at this VNTR locus correlates with steady-state levels of INS mRNA in pancreas and transfected rodent cell lines, but it is difficult to reconcile the association of lower INS mRNA levels in the pancreas with class III VNTRs that are dominantly protective from IDDM. We show that during fetal development and childhood, mRNAs for insulin and other islet cell autoantigens (GAD, ICA69, IA-2) are expressed at low levels in the human thymus. Critically, we also detect proinsulin and insulin protein. VNTR alleles correlate with differential INS mRNA expression in the thymus where, in contrast to the pancreas, protective class III VNTRs are associated with higher steady-state levels of INS mRNA expression. This finding provides a plausible explanation for the dominant protective effect of class III VNTRs, and suggests that diabetes susceptibility and resistance associated with IDDM2 may derive from the VNTR influence on INS transcription in the thymus. Higher levels of (pro)insulin in the thymus may promote negative selection (deletion) of insulin-specific T-lymphocytes which play a critical role in the pathogenesis of type-1 diabetes.

Aging↗
PubMed1997

Insulin VNTR allele-specific effect in type 1 diabetes depends on identity of untransmitted paternal allele. The IMDIAB Group.

The IDDM2 type 1 diabetes susceptibility locus was mapped to and identified as allelic variation at the insulin gene (INS) VNTR regulatory polymorphism. In Caucasians, INS VNTR alleles divide into two discrete size classes. Class I alleles (26 to 63 repeats) predispose in a recessive way to type 1 diabetes, while class III alleles (140 to more than 200 repeats) are dominantly protective. The protective effect may be explained by higher levels of class III VNTR-associated INS mRNA in thymus such that elevated levels of preproinsulin protein enhance immune tolerance to preproinsulin, a key autoantigen in type 1 diabetes pathogenesis. The mode of action of IDDM2 is complicated, however, by parent-of-origin effects and possible allelic heterogeneity within the two defined allele classes. We have now analysed transmission of specific VNTR alleles in 1,316 families and demonstrate that a particular class I allele does not predispose to disease when paternally inherited, suggestive of polymorphic imprinting. But this paternal effect is observed only when the father's untransmitted allele is a class III. This allelic interaction is reminiscent of epigenetic phenomena observed in plants (for example, paramutation; ref. 17) and in yeast (for example, trans-inactivation; ref. 18). If untransmitted chromosomes can have functional effects on the biological properties of transmitted chromosomes, the implications for human genetics and disease are potentially considerable.

Alleles↗
PubMed1997

[Perinatal outcomes of newborn infants of mothers over 40 years old. A case-control study].

The changing life patterns for women in current society are accompanied by a postponement of pregnancy in advanced reproductive age. The aim of our study was to compare perinatal outcome among 314 newborns from women > or = 40 years old with 6.683 controls from mothers 20-29-year-old who delivered at our Division between 1983 and 1993. For each analyzed variable (birth weight, gestational age, congenital malformations, still-births, neonatal mortality and morbidity) the odds ratio (OR) and 95% confidence intervals (95% CI) were calculated for the group of women > or = 40 years old. The incidence of nulliparas in the older age group was 13.5%. The cesarean section rate was higher in the mature mothers than in their younger controls (52.1% vs 34.7%). The more frequent prenatal genetic management in women past the age of 35 years may be the reason for the reduced overall frequency of still-births and malformations in the group > or = 40 years. Preterm delivery was observed more frequently in the older mothers compared with controls (18.5% vs 11.7%). The frequency of LBW and VLBW was higher, but not significant, in the cases than in the controls. Newborns from older mothers had a significant increase of macrosomia (8% vs 4.8%) and a twice increase of mortality (3.2% vs 1.6%) and morbidity (20.4% vs 11.4%). These data suggest to improve the perinatal care of women > or = 40 years old.

Adult↗
PubMed1997

Detection of HIV p24 from antigen presenting monocytes for early diagnosis of HIV-1 infection.

The monocytic/macrophagic lineage has an antigen presenting cell function also towards HIV. On the basis of this fact, a new method, indirect immunofluorescence (IIF) for measurement of p24 from monocytes was used. The results were compared to an amplified enzymatic test for serum dissociated p24 detection in 14 HIV negative individuals at risk for HIV and 12 HIV positive patients. Only one seronegative, who had a symptomatic primary HIV infection, had a positive IIF and also an elevated level of p24 in serum. The others had a negative IIF and, 6 months after the specimen, were not positive to the routine methods for detection of anti-HIV antibodies. Seronegative subjects not at risk for HIV were consistently negative to IIF. Among the HIV positive patients 4 were positive to IIF and the remaining 5 were positive to routine methods. Divergent results could be explained by the fact that one test measures cell derived antigen and the other serum antigen and that monocytes can loose APC function in the advanced stages of the illness. The test proved to be cheap and simple, and it is possible to hypothesize an application of it as a support test for the early diagnosis of HIV infection in laboratories not endowed with high levels of technology. Moreover, the results of the amplified p24 ELISA test in 44 seronegative at risk test are reported herein.

Enzyme-Linked Immunosorbent Assay↗
PubMed1996

Comparison of different in vitro tests for evaluating immune reactivity.

We performed some in vitro tests for the detection of the immune state and compared the results. In particular we studied the production of various cytokines obtained by stimulating peripheral blood mononucleated cells (PBMC) with different inducers, using optimal and suboptimal doses. This was compared with the results of blastic transformation of lymphocytes, and with the evaluation of the capping effect of macrophages, and of the Multitest Merieux. The correlation between the different investigations was generally good. This permits a simplification of the study of immune reactivity, selecting some of the tests proposed. The use of suboptimal doses of inducers improves the evaluation of very moderate deficits and supplies an in vitro model for the study of immunomodulant drugs.

Adult↗
PubMed1996

Expression of fibronectin and its receptor in some tumour cell lines and in HIV-1-infected cells.

The aim of our study was to evaluate the levels of fibronectin (FN) and its classic receptor (FNR) in various transformed cells lines, especially of leukemic origin, and also the influence of HIV-1 replication on the expression of these proteins (in particular on H9-V cells, chronically infected with HIV-1, and acutely infected MT-4 cells). Monoclonal antibodies were used for indirect immunofluorescence tests; the fluorescein-conjugated recombinant p14, the product of the HIV gene tat, was used as a molecular probe. The results demonstrated a high variability of FN and FNR expression among the various cellular lines studied. Moreover, deficits of such adhesive proteins did not necessarily correlate with a severe reduction of the corresponding receptor. HIV-1 replication in MT-4 and H9-V cells increased the expression of FNR. This seems to correlate with p14-induced phenomena because pretreatment of H9-V cells with recombinant p14 showed an enhancing effect on the expression of this receptor.

Amnion↗
PubMed1996

Influence of fibronectin on HIV-1 infection and capability of binding to platelets.

We have previously demonstrated that fibronectin (FN) can bind HIV-1 envelope proteins, in particular gp 120. The aim of the present study was to determine some biological effects of this phenomenon. Pretreating HIV-1 with human FN increased the infectivity of HIV-1, when a low concentration of the virus was used. In contrast, an RGD-containing pentapeptide (Gly-Arg-Gly-Asp-Ser), which is a fundamental binding site of FN, reduced the infectivity of a suspension of HIV-1 at high concentrations of the virus. It is likely that FN bridges the cell surface and the virions, while the RGD-containing pentapeptide may saturate the HIV-1 binding sites for cell surface receptors. Moreover, gp 120 was bound to the FN present on the surface of platelets. The specificity of this binding was confirmed by the inhibition obtained by pretreating platelets with anti-FN antibodies. The consequence of the surface modifications of the platelets could explain the thrombocytopenia that frequently occurs in patients infected with HIV and suggests also the possibility that platelets could be a vehicle for the virus in the circulation.

Blood Platelets↗
PubMed1996

Limited loss of tolerance to islet autoantigens in ICA+ first degree relatives of patients with type I diabetes expressing the HLA dqb1*0602 allele.

The DQB1*0602 allele confers dominant protection from Type I diabetes even among islet cell antibody positive (ICA+) first degree relatives of affected individuals. Lack of progression to diabetes in these subjects, despite ICA positivity, could be explained by a loss of tolerance limited to fewer islet autoantigens than in ICA+ relatives progressing to the disease. To test this hypothesis we have determined autoantibodies by radioassay to three islet autoantigens, glutamic acid decarboxylase (GAD), insulin and the novel neuroendocrine antigen ICA512/IA-2 in 84 HLA-typed ICA+ first degree relatives of patients with Type I diabetes. Among the eleven relatives expressing the 0602 allele (0602+) only two were positive for more than one antibody as determined by radioassay. In contrast, 55/73 ICA+ relatives lacking this allele (non-0602) expressed more than one autoantibody (P < 0.01). The prevalence of antibodies to GAD (GAA) was not significantly different in ICA+ relatives with and without the 0602 allele (7/11 in 0602+ versus 60/73 in non-0602). In contrast, anti-insulin antibodies (IAA) were present in only 2/11 0602+ ICA+ relatives versus 47/70 in non-0602 ICA+ relatives (P < 0.01). Similarly, only one individual carrying the 0602 allele was positive for ICA512 autoantibodies versus 33/70 in the non-0602 group (P < 0.01). These data indicate that even among ICA+ relatives the 0602 allele is associated with a limited immune response to islet antigens and amongst 0602+ relatives this response is mostly directed against GAD.

Adolescent↗
PubMed1996

Effects of recombinant murine (rm) interleukin-12 and rm interferon-gamma in mice infected with Bordetella pertussis.

The recombinant cytokines interferon (IFN)-gamma and interleukin (IL)-12 stimulate several macrophage-mediated functions that are important in host defense. An experimental pertussis model showed that intraperitoneal (i.p.) administration of 10,000 U of recombinant murine (rm) IFN-gamma to mice at the time of Bordetella pertussis infection caused a marked and significant reduction in the number of colony-forming units of bacteria in the lungs. Administration i.p. of 1 microgram of rmIL-12 or 1 microgram of rmIL-12 at the time of and for 5 consecutive days after B. pertussis challenge also induced a significant reduction in the number. However, i.p. administration of 1 microgram of rmIL-12 with 10,000 U of IFN-gamma at the time of B. pertussis challenge did not provide protection. These findings indicate that exogenous administration of rmIL-12 and rmIFN-gamma enhances resistance of mice to B. pertussis infection.

Animals↗