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Biomedical subjects

J Prieto

Publications and source records attributed to J Prieto.

At least 235 records · Page 13Linked to original sources
PubMed1994

Regulated expression of integrins and other adhesion molecules during differentiation of monocytes into macrophages.

Monocyte/macrophages adhere to cells (lymphocytes, vascular endothelial and other cell types) and to extracellular matrix components (fibronectin and laminin) by using specific cell surface adhesive structures. In the present study we have analyzed expression of integrins, immunoglobulin (Ig)-related, selectins, and other adhesion molecules on blood monocytes, in vitro differentiated macrophages (ivMs), and alveolar macrophages (AMs), obtained from healthy nonsmokers by bronchoalveolar lavage (BAL). We have also investigated expression of adhesion molecules on myelomonocytic cell lines HL-60, THP-1, KG-1, and U937 before and after tetradecanoyl phorbol acetate (TPA)-induced differentiation. With regard to the integrin family, monocytes expressed beta 1 (CD29), alpha 4, alpha 5, alpha 6, beta 2 (CD18), CD11a, CD11b, and CD11c subunits, but not alpha V (CD51). Some reactivity with mAbs against the platelet antigens CD41b (IIb) and CD61 (beta 3) was detected. The Ig-related molecules CD54 (ICAM-1), ICAM-2, and CD58 (LFA-3) were expressed, as well as L-selectin and the carbohydrate ligands Le(x) (CD15) and sialyl Le(x). Immunolabeling for the structurally unrelated molecules CD44 and CD36 was strongly positive. In comparison to monocytes, AMs showed much lower expression of alpha 4, alpha 6, beta 2, CD11a, CD11b, L-selectin, Le(x), and sialyl Le(x). Moreover, ICAM-2 and CD36 were practically absent whereas expression of alpha 3, but not of CD11c, was higher. Similar results were obtained with ivMs. All four myelomonocytic cell lines showed down-regulation of alpha 4 and up-regulation of CD11c after TPA treatment. These findings indicate that maturation of monocytes into macrophages is accompanied by characteristic changes in adhesion molecule expression. The particular array of adhesion molecules on monocytes and macrophages may account for differences in the functional properties of these cells.

Antigens, CD↗
PubMed1994

Overcoming class II-linked non-responsiveness to hepatitis B vaccine.

This work shows that class II-linked humoral lack of response to an antigen can be overcome by joint immunization with the antigen and a T-helper cell determinant (TDh) well recognized by class II molecules of a non-responder individual. Thus, SJL/J mice (H-2s), which are non-responders to the S region of hepatitis B virus surface antigen (HBsAg), were rendered responders by joint immunization with a recombinant surface antigen, only composed of the S region, and a short synthetic TDh peptide well recognized by the H-2s restriction. By contrast, when this peptide is not recognized as TDh, as in B10M mice (H-2f restricted and also non-responders to the S region), no humoral response could be induced against the S region. These results have important implications for therapy and vaccination against hepatitis B virus as well as in enhancing the immunogenicity of other antigens.

Amino Acid Sequence↗
PubMed1994

Atrial natriuretic factor in cirrhosis: relationship to renal function and hemodynamic changes.

Plasma atrial natriuretic factor concentrations and different hemodynamic parameters, including the evaluation of femoral arteriovenous shunting by measuring the arteriovenous difference of oxygen content (Ca-vO2), were determined in eight healthy subjects and 24 patients with cirrhosis without renal failure (group I: seven patients without ascites, group II: nine patients with ascites and UNaV > 10 mEq/24 h and group III: eight patients with ascites and UNaV < or = 10 mEq/24 h). Atrial natriuretic factor was 34 +/- 4.7 pg/ml in the control group and 44.28 +/- 5.4, 67.89 +/- 8.8 and 84 +/- 10.8 pg/ml in groups I, II and III respectively (p < 0.001. group III vs. I and control and II vs. control). Atrial natriuretic factor directly correlated with cardiac index (p < 0.01), blood volume (p: 0.01), femoral blood flow (p < 0.01) and inversely with systemic and femoral vascular resistances (p < 0.02), Ca-vO2 (p < 0.01), serum albumin (r: -0.61; p < 0.01) and prothrombin index (r: -0.63; p < 0.02). These results indicate that plasma atrial natriuretic factor is increased in patients with cirrhosis, especially in those with advanced disease and marked renal sodium retention. This suggests that in cirrhosis, arteriolar vasodilation and peripheral arteriovenous shunting influence renal function while inducing a state of overflow at the central venous compartment leading to increased atrial natriuretic factor secretion. Increased production of this vasodilatory hormone may thus contribute to the hyperkinetic circulation of cirrhosis.

Atrial Natriuretic Factor↗
PubMed1994

Prediction of sustained remission of chronic hepatitis C after a 12-month course of alfa interferon.

alpha-Interferon therapy normalizes aminotransferase levels in approximately 50% of the patients with chronic hepatitis C, but post-therapy relapses are common and predictive factors of sustained response remain largely unknown. We retrospectively assessed several parameters as predictors of sustained remission after a 12-month course of lymphoblastoid alpha-interferon: the Knodell histological activity index, serum levels of procollagen type III peptide, serum HCV-RNA, anti-alpha-interferon antibodies, and anti-HCV antibodies (C-100-3), all at month 12. Thirty-seven patients were studied. Fourteen patients were non-responders (38%), 15 patients experienced a sustained response (40.5%) and eight patients responded similarly but relapsed after alpha-interferon withdrawal (21.5%). A decrease in the histological activity index above 5, normalization of procollagen type III peptide levels (< 12 ng/ml) and the absence of viremia after treatment were all significantly associated with a sustained response (p = 0.008, p = 0.007 and p = 0.037, respectively). Anti-interferon antibodies were detected in only one non-responder patient. Anti-C-100-3 antibodies became undetectable at month 12 in 5 of the 15 sustained responders. The best prediction of sustained response was obtained from the three variables independent of multivariate analysis according to the following equation: F = 0.872 + 0.067 x K (decrease of histological index) -0.052 x P (procollagen type III peptide levels at month 12) -0.28 x R (HCV-RNA at month 12; R = 2 when present and R = 1 when absent). A score higher than 0 predicted sustained remission with a 100% sensitivity and specificity in this series of patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Activity of fluconazole: postantifungal effect, effects of low concentrations and of pretreatment on the susceptibility of Candida albicans to leucocytes.

Exposure times and serum concentrations in humans after treatment with multiple doses of fluconazole have been studied to measure: (a) the postantifungal effect (PAFE) on Candida albicans, at two concentrations of the drug in the presence or absence of 10% human serum; (b) the activity of low concentrations of the drug on yeasts previously exposed to fluconazole with or without 10% human serum; and (c) the effect of fluconazole pretreatment on the fungicidal activity of leucocytes and serum against C. albicans. Fluconazole showed no PAFE against C. albicans between -0.1 and -0.7 h, but when the assays were performed in the presence of serum, concentration dependent PAFEs were obtained (1.1 h-3.6 h). Pretreated yeasts were more susceptible than untreated yeasts to low concentrations of the drug. The decrease in growth was dependent on the concentration used in pretreatment. Growth delay was also more marked when the yeast cells were tested in the presence of serum. Pretreatment of the growing C. albicans cells with fluconazole increased their vulnerability to killing by leucocytes mainly in the first hour (P < 0.05). These results could partially explain why fluconazole has more activity in vivo than in vitro.

Blood Physiological Phenomena↗
PubMed1994

Immunohistochemical detection of chloride/bicarbonate anion exchangers in human liver.

Sodium-independent Cl-/HCO3- exchange activity has been observed in isolated rat hepatocytes and intrahepatic bile duct epithelial cells, where it is involved in intracellular pH regulation and, possibly, biliary bicarbonate secretion. Monoclonal antibodies to the membrane domain of human chloride/bicarbonate anion exchanger proteins, AE1 and AE2, were prepared so that we might determine by immunohistochemical methods the presence and location of these antiporters in the human liver. To obtain the antibody against AE1, we immunized mice with injections of washed human erythrocytes. The selected monoclonal antibody was found to be specific for the 17-kD proteolytic membrane fragment of AE1 protein. The antibody to AE2 was produced with a 14-mer synthetic peptide, whose sequence corresponds specifically to amino acid residues 871 to 884 in the deduced primary structure of human kidney AE2 protein. When the monoclonal antibody to AE2 peptide was employed for the immunohistochemical study of liver specimens (by both immunofluorescence and immunoperoxidase), a clearly defined staining was present at the canalicular membrane of hepatocytes, as well as the luminal side of the membrane of bile duct epithelial cells from small and medium-sized bile ducts. No staining was observed in the liver parenchyma with the monoclonal antibody to AE1, which instead strongly decorated the erythrocytes in liver blood vessels. We conclude that AE2 immunoreactivity is present in human liver, where it localizes very specifically to the membrane regions, which appear most probably involved in the transport of bicarbonate to bile (i.e., the canalicular membrane of hepatocytes and the apical side of epithelial cells of small and medium bile ducts).

Amino Acid Sequence↗
PubMed1994

Induction of neutralizing antibodies against human immunodeficiency virus type 1 using synthetic peptide constructs containing an immunodominant T-helper cell determinant from vpr.

Identification of immunodominant T-helper-cell determinants after natural infection is an important step in the design of immunogens for potential use in vaccination. Using cells from human immunodeficiency virus type 1 (HIV-1)-infected individuals and a panel of peptides encompassing the sequence of the regulatory protein vpr from HIV-1, we identified the T-helper determinant QLLFIHFRIGCRHSR, which is active in 37.5% of these individuals. To gain insight on the efficacy of this peptide in helping induce neutralizing antibodies against a B-cell determinant (BD), we synthesized constructs containing B- and T-cell determinants and tested them in BALB/c mice, the highest responders to the T-cell determinant moiety among several strains tested. These immunogens induced antibodies against two chosen B-cell determinants from HIV-1IIIB gp160 (amino acids 310-322 from the V3 loop of gp120 and 736-751 from gp41) that were able to neutralize HIV-1 infection in vitro. The highest neutralization titer against HIV-1IIIB was obtained by immunization with the homopolymer of the construct containing the T-cell epitope from vpr and the B-cell epitope from the V3 loop. We believe that the immunodominant T-cell determinant from vpr is a promising epitope to consider in the design of future peptide vaccines.

Amino Acid Sequence↗
PubMed1994

CD4-modified synthetic peptides containing phenylalanine inhibit HIV-1 infection in vitro.

Phenylalanine-containing peptides from CD4 were synthesized based on chemical similarity with active CD4(81-92)-benzylated peptides. The synthetic peptide FYIFFVEDQKEEDD blocked the binding of gp120 to CD4 and inhibited 50% human immunodeficiency virus (HIV)-induced syncytia formation at a concentration (IC50) of approximately 40-50 microM and HIV p17 expression with an IC50 of approximately 67 microM. The peptide is not toxic to cells in vitro. Moreover, acute toxicity studies carried out in Swiss mice showed the peptide to be nontoxic at a dose of 2,000 mg/kg. This phenylalanine-substituted CD4 peptide may prove to be useful in the treatment of AIDS.

Amino Acid Sequence↗
PubMed1993

Thermodynamic analysis of the chemotactic protein from Escherichia coli, CheY.

CheY, the 129 amino acid chemotactic protein from Escherichia coli, is a good model for studies of folding of parallel alpha/beta proteins. We report here the thermodynamic characterization of the wild-type CheY at different pH values and in different buffers and denaturation conditions. The denaturation of CheY by urea monitored by circular dichroism and fluorescence fits the two-state unfolding model. The stability of the protein is ionic strength dependent, probably due to the presence of three Asp residues in very close proximity in its active site. The presence of a Mg2+ ion, which seems to interact with Asp 13 in the active site, stabilizes the native structure by up to 6.9 kJ mol-1. The CheY maximum stability (31.7 +/- 2.1 kJ mol-1), without magnesium, is reached at pH 5.1. Analysis of scanning calorimetry data has shown that temperature-induced unfolding of CheY is not a two-state process and proceeds through a highly populated intermediate state, corresponding to protein dimers, as was subsequently confirmed by direct cross-linking experiments. According to circular dichroism, fluorescence, nuclear magnetic resonance, and ANS binding experiments, this "intermediate dimer" at pH 2.5 exhibits all known characteristics of the "molten globule" state. The reversible dimerization of "molten globules" might explain such peculiarities as the increased stability or the cooperative unfolding found for the molten globule state of some proteins.

Anilino Naphthalenesulfonates↗
PubMed1993

1H- and 15N-NMR assignment and solution structure of the chemotactic Escherichia coli Che Y protein.

Che Y is a 129-residue parallel alpha/beta protein involved in bacterial chemotaxis. We have used this protein as a model to study the folding reaction of parallel alpha/beta proteins. As a first step we carried out the complete assignment of the 1H and 15N spectra from Escherichia coli Che Y protein on the basis of two-dimensional 1H homonuclear and 1H-15N heteronuclear experiments by using sequence-specific methods. Our assignments differ from the preliminary assignments made by Kar et al. [Kar, L., Matsumura, P. & Johnson, M.E. (1992) Biochem. J. 287, 521-531] of aromatic residues obtained by comparison of NOEs with short proton-proton distances in the crystal structure of Che Y. The analysis of the extension of the secondary elements, as well as a preliminary calculation of the three-dimensional structure, indicate that the solution structure is closely coincident with the single crystal structure determined by X-ray diffraction.

Amino Acid Sequence↗
PubMed1993

Arteriovenous shunting, hemodynamic changes, and renal sodium retention in liver cirrhosis.

BACKGROUND: Arterial vasodilation has been postulated to initiate sodium retention in cirrhosis. This work was designed to analyze whether arteriovenous shunting underlies vasodilation and influences renal function in cirrhosis. METHODS: The femoral arteriovenous difference in oxygen content (Ca-VO2) was measured in 10 healthy subjects (control group) and 31 cirrhotic patients: 9 without ascites (group 1), 10 with ascites and a urinary sodium excretion rate (UNaV) of > 10 mEq/24 h (group 2), and 12 with ascites and UNaV of < or = 10 mEq/24 h (group 3). In 8 subjects from each group, femoral blood flow and the cardiac output were determined by duplex-Doppler ultrasonography. In 9 cases arteriovenous shunting in the femoral territory was estimated using 30 +/- 5-microns radiolabeled microspheres. RESULTS: Ca-VO2 was lower in group 3 than in controls or group 1. Ca-VO2 correlated inversely with femoral blood flow, plasma renin activity, plasma aldosterone concentration, and degree of shunting measured by microspheres. Ca-VO2 correlated directly with systemic vascular resistance and prothrombin index. CONCLUSIONS: In decompensated cirrhotic patients, there is an increased arteriovenous shunt for oxygen in the lower extremities that is associated with increased arterial blood flow, decreased systemic vascular resistance, and worsening of liver function. This shunt is due partly to opening of arteriovenous precapillary connections.

Aldosterone↗
PubMed1993

Abnormal expression of anion exchanger genes in primary biliary cirrhosis.

BACKGROUND: Chloride-bicarbonate anion exchangers (AE) are thought to be essential to the generation of ion fluxes into biliary, salivary, pancreatic, and other secretions. In primary biliary cirrhosis (PBC), chronic cholestasis is frequently associated with pluriglandular exocrine failure. The present study examined the expression of AE genes in PBC at the messenger RNA (mRNA) level. METHODS: Two AE messages, i.e., AE1 and AE2 mRNAs, were analyzed by a polymerase chain reaction-based procedure in liver specimens and peripheral mononuclear cells of patients with PBC and control subjects (with normal livers or with miscellaneous liver diseases). RESULTS: AE1 mRNA was detected in only a few liver biopsy specimens of patients with PBC and control subjects, whereas AE2 mRNA was found in all liver specimens tested. In lymphoid cells, however, both transcripts were consistently detected. AE2 mRNA levels in liver as well as in lymphoid cells were significantly lower in PBC patients than in controls. In contrast to AE2 mRNA, AE1 mRNA levels were increased in lymphoid cells from patients with PBC. Interestingly, the levels of liver AE2 mRNA in a few PBC patients treated with ursodeoxycholic acid were within the range of the controls. CONCLUSIONS: The expression of AE genes is altered in patients with PBC. It is suggested that this disturbance may have a role in the cholestasis, exocrine failure, and immunodysfunction of PBC.

Adult↗
PubMed1993

Liver cytoprotection by prostaglandins.

During the last decade intensive work on the relationships between the liver and the arachidonic acid cascade has greatly expanded our knowledge of this area of research. The liver has emerged as the major organ participating in the degradation and elimination of arachidonate products of systemic origin. The synthesis in the liver of arachidonate products derived from the cyclooxygenase, lipoxygenase and cytochrome P450 system pathways has been demonstrated. The participation of leukotriene B4 and cysteinyl-leukotrienes as mediators of liver damage and the possible therapeutic usefulness of prostaglandins (PGs) in acute liver injury has attracted the interest of clinicians. This article reviews the essential features regarding the role of arachidonate metabolites in liver disease and specially focuses on the cytoprotective effects on the liver displayed by PGE2, PGE1, PGI2 and synthetic PG analogs in experimental models of liver damage induced by ischemia-reperfusion injury, carbon tetrachloride, bacterial lipopolysaccharide and viral hepatitis and on the possible mechanisms underlying liver cytoprotection in these experimental models. The therapeutic usefulness of PGs in clinical practice is critically analyzed on the basis of available evidence in patients with fulminant hepatic failure and primary graft nonfunction following liver transplantation.

Animals↗
PubMed1993

Graft-versus-host disease after liver transplantation.

We describe a case of graft-versus-host disease that developed in the recipient of an orthotopic liver transplant. A maculopapular eruption developed on postoperative day 25. After the diagnosis of graft-versus-host disease was made aggressive therapy was instituted, which resulted in a complete recovery.

Exanthema↗
PubMed1993

Enhancement of peptide immunogenicity by insertion of a cathepsin B cleavage site between determinants recognized by B and T cells.

The insertion of two lysine residues (cleavage sites of cathepsin B) at the boundary of a peptide recognized by B cells (BD) and a class-II- presentable sequence (TDh) enhanced the anti-BD antibody induction capacity of this type of peptide construct, as well as production of IL2. It is postulated that these lysines generate a neoprocessable site which helps in release of the TDh moiety from the construct, enabling its presentation to class II molecules, an essential step in clonal expansion of the antibody-producing B cell after internalization of the construct via the BD moiety.

Amino Acid Sequence↗
PubMed1993

Detection of anti-hepatitis C virus antibodies by ELISA using synthetic peptides.

A novel ELISA assay for the detection of anti-hepatitis C virus antibodies in the sera of infected individuals is described. This assay is based on a mixture of three 15-amino acid synthetic peptides encompassing regions of core and NS4 proteins of hepatitis C virus. Comparison with other available ELISA assays based on recombinant polypeptides shows that, short synthetic peptides have the advantage over some larger recombinant peptides by giving higher specificity without loss of sensitivity.

Enzyme-Linked Immunosorbent Assay↗
PubMed1993

Hepatitis B virus occult infection in subjects with persistent isolated anti-HBc reactivity.

The aim of this study was to investigate the presence of hepatitis B virus occult infection in asymptomatic subjects with persistent anti-HBc reactivity but no other hepatitis B virus serological markers, including HBsAg, anti-HBs, IgM anti-HBc and HBV-DNA. For this purpose we used both polymerase chain reaction assays in sera and immunohistochemistry for HBsAg and HBcAg in liver biopsy specimens. Twenty-four cases were studied: 15 were drug abusers or homosexuals (eight with normal alanine aminotransferase levels) and nine were heterosexuals with raised alanine aminotransferase levels (> 45 U/l) but with no history of blood transfusion or ethanol intake (< 80 g daily). In all but five cases, liver biopsy was performed in subjects with persistent elevated alanine aminotransferase levels. In 10 out of 24 cases (41.66%) hepatitis B virus infection was demonstrated by polymerase chain reaction or immunohistochemistry, and when results from both procedures were available (n = 11) hepatitis B virus infection was detected in 63.63% of the subjects. The only clinical feature associated with HBV infection was the presence of persistent elevated alanine aminotransferase levels (p < 0.05). In conclusion, persistent isolated anti-HBc reactivity may be a relatively common serologic pattern for hepatitis B virus occult infection, at least in patients with chronic liver disease.

Base Sequence↗