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Biomedical subjects

J Prieto

Publications and source records attributed to J Prieto.

At least 199 records · Page 11Linked to original sources

Tumor necrosis factor alpha gene expression and the response to interferon in chronic hepatitis C.

Tumor necrosis factor alpha (TNF-alpha) is a cytokine with pleiotropic properties that is induced in a variety of pathological situations including viral infections. In this work, we analyzed the expression of TNF-alpha gene in patients with chronic hepatitis C. Serum TNF-alpha levels were found to be elevated in all chronic hepatitis C patients including those cases presenting sustained biochemical remission of the disease after interferon therapy. Untreated patients with chronic hepatitis C showed increased TNF-alpha messenger RNA (mRNA) levels in the liver and mononuclear cells as compared with healthy controls. After completion of treatment with interferon, patients experiencing sustained complete response showed values of TNF-alpha mRNA, both in the liver and in peripheral mononuclear cells, within the normal range, significantly lower than patients who did not respond to interferon and than those with complete response who relapsed after interferon withdrawal. Pretreatment values of TNF-alpha mRNA were lower in long-term responders to interferon than in cases who failed to respond to the treatment. Values of TNF-alpha mRNA in the liver or in mononuclear cells were higher in specimens with positive hepatitis C virus (HCV) RNA than in those samples where the virus was undetectable. Neither the intensity of the liver damage nor the amount of HCV RNA in serum or in cells showed correlation with the levels of TNF-alpha transcripts in peripheral mononuclear cells but it was found that high TNF-alpha values were associated with genotype 1b. In conclusion, there is an enhanced expression of TNF-alpha in HCV infection. High levels of this cytokine may play a role in the resistance to interferon therapy.

Adult↗

Epidemiological, clinical and therapeutic associations of hepatitis C types in western European patients.

BACKGROUND/AIMS: Different variants of hepatitis C virus might show different susceptibility to interferon alpha treatment, but it is important to understand whether this difference in sensitivity reflects an association with other factors, such as cirrhosis or age. METHODS: We have used an enzyme-linked immunosorbent hepatitis C virus typing assay based upon the detection of antibody in patient's era to type-specific NS-4 antigens to investigate the effect of hepatitis C virus type in 610 patients with chronic hepatitis C virus infection. The influence of viral types and their interdependency with host factors were separately analyzed to establish which factors executed an independent effect on the probability of sustained response. RESULTS: There was a marked difference in the distribution of hepatitis C virus types with age: infection with type 3 was more common in younger patients. The distribution of hepatitis C virus type with age is accounted for by differences in risk-factors for infection in different age groups. The frequency of cirrhosis increased markedly with age. Even after standardization for age, center and the presence of cirrhosis, viral type was strongly related to the outcome of infection. CONCLUSIONS: Our data suggest that enzyme-linked immunosorbent hepatitis C virus typing could assist in patient selection for interferon treatment to improve sustained response rates. Together with measurement of viral load, hepatitis C virus typing may serve to indicate the probability of response in patients with chronic hepatitis C, and to elucidate antiviral mechanisms in the disease. The serotyping assay provides a relatively inexpensive screening method to determine the infecting hepatitis C virus type, which could facilitate therapeutic decisions and strategies in patients with chronic hepatitis C.

Adolescent↗

Production of interleukin-2 in response to synthetic peptides from hepatitis C virus E1 protein in patients with chronic hepatitis C: relationship with the response to interferon treatment.

BACKGROUND/AIMS: The role of cellular immunity in the clearance of hepatitis C virus after interferon therapy has not yet been elucidated. Here, we analyzed the T cell response to peptides from hepatitis C virus E1 protein in untreated and interferon-treated patients with chronic hepatitis C virus infection. METHODS: We used thirty-six 15-mer synthetic peptides from hepatitis C virus E1 protein (genotype 1a) in a sensitive interleukin-2 production assay in two groups of controls (healthy seronegative individuals and patients with liver diseases unrelated to hepatitis C virus), and three groups of patients with chronic hepatitis C: nine patients who cleared the virus after interferon treatment (group 1), nine patients who failed to respond to the therapy (group 2) and nine previously untreated patients (group 3). RESULTS: None of the controls responded to any of the peptides tested, whereas 8/9 (88%) of patients from group 1 responded positively. In contrast, only 2/9 (22%) of patients from group 2 showed peptide recognition. In group 3, 5/9 patients (55%) displayed positive response against E1 peptides. When E1 peptides from the sequence corresponding to genotype 1b (the commonest in patients who were non-responders to interferon) were tested in nine additional interferon-resistant patients (group 2*) a positive response was detected in only three of them (33%). CONCLUSIONS: T cell recognition of hepatitis C virus E1 peptides in patients with chronic hepatitis C who exhibit sustained response to interferon therapy is increased as compared with interferon-resistant cases, suggesting that T cell immunity to hepatitis C virus structural proteins may play a role in the clearance of this viral infection.

Adult↗

Hepatology.

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Ascites↗

Serum angiotensin converting enzyme and C4 protein of complement as a combined diagnostic index in alcoholic liver disease.

Diagnosis of liver cirrhosis relies on hepatic biopsy. So far, attempts have failed to achieve a serologic test that differentiates cirrhosis from other hepatic conditions. The aim of this work was to assess the diagnostic value of the ratio of serum angiotensin converting enzyme activity (SACE) and the levels of protein C4 of serum complement (SACE/C4) in differentiating cirrhotic from noncirrhotic alcoholic liver diseases. In this study, 68 active alcoholic patients (17 with fatty liver or minimal changes, 11 with acute alcoholic hepatitis and 40 with cirrhosis) were included. Twenty healthy subjects were studied as a control group. Liver biopsy was performed in all patients. SACE levels were significantly higher in the group with cirrhosis when compared with the group of patients without cirrhosis and the control. On the other hand, serum C4 level decreased as liver damage progressed. SACE values above 25 IU/l had a sensitivity of 92.5 percent (95 percent confidence interval, 87.5 to 97.5) and a specificity of 79 percent (95% percent confidence interval, 70.5 to 87.5), in detecting those patients with liver cirrhosis. The sensitivity further increased to 95 percent (95 percent confidence interval, 90.5 to 99.5) and the specificity to 100 percent when the SACE/C4 ratio was used and a cutoff point of 145 was chosen. To conclude, in alcoholics SACE is specifically elevated in patients with cirrhosis, and the SACE/C4 ratio is a excellent biochemical index for the diagnosis of cirrhosis in alcoholic patients.

Adult↗

Ex vivo antibacterial properties of rufloxacin compared with those of norfloxacin in a study with healthy volunteers.

Twelve adult males participated in a randomized crossover phase I clinical trial comparing serum bactericidal titers (SBTs), urine bactericidal titers (UBTs), and urine killing rates (UKRs) against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213, after the administration of single 400-mg doses of rufloxacin and norfloxacin at different times up to 72 h postdose. SBTs were significantly higher (P < 0.05) against E. coli from 8 to 48 h and against S. aureus from 4 to 24 h with rufloxacin. UBTs for E. coli were higher (P < 0.05) for norfloxacin at early sample times (0 to 8 h) but higher for rufloxacin (P < 0.05) at sample times from 16 h on for both E. coli and S. aureus. Similar UKRs were obtained for both quinolones for 0 to 2 h and 8 to 12 h, but the UKR was maintained for 72 h with rufloxacin. The high and sustained mean levels of rufloxacin in urine (> 35 micrograms/ml), median UBTs (> 32 for E. coli and 16 for S. aureus) and UKRs for E. coli suggest prolonged urine antibacterial activity (for at least 72 h) and its use as a single 400-mg dose in the treatment of uncomplicated cystitis.

Adult↗

Activities against Streptococcus pneumoniae of amoxicillin and cefotaxime at physiological concentrations: in vitro pharmacodynamic simulation.

An in vitro model simulating amoxicillin and cefotaxime concentrations in human serum (after standard doses) was used to explore the activities of these drugs over time against penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae strains. An initial inoculum reduction percentage of > or = 90% was obtained with amoxicillin and maintained for 2 to 8 h, regardless of the strain tested. In contrast, experiments showed that cefotaxime had significantly (P < 0.001) less capability to reduce initial inocula of the penicillin-resistant pneumococci from 0.5 h on than amoxicillin, despite the same in vitro susceptibility to amoxicillin and cefotaxime in both strains.

Amoxicillin↗

Leukaemia of natural killer cell large granular lymphocyte type with HLA-DR-CD16-CD56bright+ phenotype.

The case is reported of a 45 year old woman with the rare leukaemia of natural killer cell large granular lymphocyte (NK/ LGL) type. Cytometric analysis of leukaemic blasts showed that they were positive for CD2, CD38, and CD56 antigens but negative for a series of antigens including CD3, CD7, CD16, and HLA-DR. Rearrangements of the beta T cell receptor, and heavy and kappa immunoglobulin genes were not detected and neither were chromosomal abnormalities. Leukaemic blasts developed NK cytotoxicity. The patient failed to respond to aggressive chemotherapy and died three months after diagnosis. The lack of expression of HLA-DR is an extraordinary characteristic of this case, as all cases of acute NK cell leukaemias described to date expressed HLA-DR. The immunophenotype observed in the NK cell leukaemic blasts may represent the counterpart of a hypothetical normal cell precursor in an early stage of ontogenic NK cell development.

Acute Disease↗

Postantifungal effect and effects of low concentrations of amphotericin B and fluconazole on previously treated Candida albicans.

The objective of the study was to assess (a) the postantifungal effect (PAFE) of amphotericin B (AmB) and fluconazole (Flu) on two Candida albicans strains, and (b) the effect of low concentrations of AmB or Flu on yeasts previously exposed to AmB or Flu, respectively, in the presence or absence of 10% human serum. In the absence of serum, AmB exerted a positive effect (at 0.5-5.0 h) depending on the concentration and the strain used. Flu, however, produced negative effects (at -1.2 to -0.1 h). When the experiment was conducted in the presence of serum, the duration of all PAFEs increased significantly, especially those induced by Flu that became positive. Pretreated yeasts were more susceptible than untreated controls to the antifungal activity of low concentrations of the drugs. Growth delay was more pronounced with Flu (up to 5.2 h), whereas the greatest decrease in log10 at 12 h was slightly more pronounced for AmB. A knowledge of PAFEs and the effects of low drug concentrations on pretreated yeasts and the effects of serum on these is important in order to gain more insight into the in vivo activity of the study drugs.

Amphotericin B↗

Circadian locomotor activity rhythm during ontogeny in crayfish Procambarus clarkii.

The characteristics of the circadian rhythm of locomotor activity in the crayfish Procambarus clarkii during ontogeny under constant darkness and light-dark (LD 12:12) conditions were studied in 132 juvenile crayfish, aged 10-140 days, divided in four groups. All animals were individually monitored with a motor activity recording system. Activity was quantitatively and qualitatively analyzed. All ages showed a circadian rhythm, although the probability of its appearance increased with age. Period values oscillated between 25.0 h in group I (2-4-week-old animals) and 24.3 h in group IV (16-20-week-old animals with more than 6 molts), but always with a high standard deviation. Groups II (5-10-week-old animals) and IV showed a statistically significant bimodal nonrandom synchrony of phases. The activity/ rest relationship diminishes as development progresses and is most uniform in group IV. We discuss the possibility that the pacemaker system responsible for this rhythm might be present from the moment of eclosion, but the coupling strength of this system with the effectors might change along development. The results presented in this work seem to indicate that the central pacemakers responsible for the activity and the ERG rhythm are not the same.

Animals↗

[Sympathetic nervous activity in cirrhosis: the relationship between peripheral hemodynamics and renal function changes].

The relationship between the activity of the sympathetic nervous system and the systemic and peripheral hemodynamic and renal function changes and the antinatriuretic activity in a group of 30 cirrhotic patients and 8 healthy subjects (control group) was investigated. Plasma catecholamines (noradrenaline, adrenaline and dopamine), plasma renin activity, plasma aldosterone concentrations, mean arterial pressure, cardiac output, blood volume, right femoral arterial flow (RFAF) and renal function parameters were determined in all the participants. Cyclic GMP urinary excretion (cV-UGMP) was used as an indirect index of systemic nitric oxide production. The plasma concentration of noradrenaline was greater in the patients than in the control group and was directly correlated with other vasopressor and antinatriuretic systems, RFAF, cV-UGMP, the degree of hepatic failure studied by the Pugh score and was inversely correlated with creatinine clearance and urinary sodium excretion. On Cox regression analysis, only the RFAF, creatinine clearance and plasma adrenaline concentration remained independently associated with plasma noradrenaline levels. Furthermore, plasma noradrenaline was significantly higher in patients with greater hyperdynamic circulation. These results indicate that extrasplanchnic vasodilatation substantially contributes to sympathetic nervous hyperactivity which may significantly influence in the renal function changes observed in these patients.

Creatinine↗

A potential experimental model for the study of osteopenia in CCl4 liver cirrhotic rats.

In order to search for an experimental model to further investigate the osteopenia associated to liver cirrhosis (LC), this study has been focused on investigating the occurrence of bone disorders in male rats to which LC histologically confirmed was induced through the validated procedure of CCl4 inhalation. Length, anteroposterior and lateromedial diameters, densitometry, mechanical stress resistance, hydroxyproline (OHprol) and calcium and phosphate contents were measured in femurs from control (n = 10) and liver cirrhosis rats (n = 10). It has been found that femurs from liver cirrhosis rats showed a significant reduction (p < 0.01) in bone weight (0.254 +/- 0.003 vs 0.230 +/- 0.004 g/100 g b.w.), anteroposterior (4.08 +/- 0.06 vs 3.69 +/- 0.05 mm) and lateromedial (5.33 +/- 0.05 vs 5.08 +/- 0.04 mm, p < 0.05) diameters, resistance to mechanical stress (405.8 +/- 9.5 vs 332.5 +/- 9.1 N) and total densitometry (0.416 +/- 0.005 vs 0.381 +/- 0.004 g/cm2). However, no significant differences were observed in bone length, calcium, OHprol and phosphate (all expressed as mg/100 mg fresh bone tissue) contents. Therefore, the proteins matrix to mineral contents ratio was not altered. These results indicate that in this model of experimental liver cirrhosis there is osteopenia characterized by bone frailty and reduced thickness, and it could offer an experimental model to study bone changes associated to liver cirrhosis.

Animals↗

Cloning and expression of the Mycobacterium fortuitum superoxide dismutase gene.

In this paper we report the cloning, sequencing and expression of the superoxide dismutase (sod) gene from Mycobacterium fortuitum. A single gene was found to code for superoxide dismutase activity with its identity being confirmed by expression in M. aurum. The amino acid sequence was found to be similar to that of superoxide dismutases of several other origins. A region downstream of the sod gene also showed similarities to the corresponding sequences of the two main mycobacterial pathogens: M. leprae and M. tuberculosis. Analysis of enzymatic activity showed this enzyme in M. fortuitum required manganese as cofactor.

Base Sequence↗

Heterologous expression of the highly conserved acidic ribosomal phosphoproteins from Dictyostelium (changed from Dictyosteliumm) discoideum in Saccharomyces cerevisiae.

The genes encoding the acidic ribosomal phosphoproteins DdP1 and DdP2 from Dictyostelium discoideum have been cloned into yeast plasmid vectors under the control of the inducible GAL1 promoter. These constructions have been used to transform S. cerevisiae strains D45 and D67 lacking the equivalent ribosomal components. The D. discoideum genes are properly transcribed when cells are grown in the presence of the inducer galactose and the mRNAs incorporated into polysomes. However, the heterologous ribosomal proteins are not able to rescue the growth deficiency in S. cerevisiae caused by the absence of their own ribosomal proteins. When the heterologous proteins are analyzed using specific antibodies, only protein DdP1 is found in the ribosomes of the transformed S. cerevisiae D67 strain. No other heterologous protein is found in any other transformed strain, suggesting that the heterologous acidic ribosomal components are rapidly degraded when they are not bound to the ribosomes. The results indicate that D. discoideum DdP1 protein is able to interact with the yeast ribosome, though the interaction is functionally inefficient. Protein DdP2, in spite of having a higher sequence similarity to its yeast counterparts, is completely inactive in S. cerevisiae. Since the P proteins from both organisms have extensive amino acid sequence similarity ranging from 60% to 70%, these results warns about establishing a direct relationship between the extent of amino acid sequence similarity and the capacity of heterologous proteins to be functional in host species. Moreover, our data suggest that evolution affected the interaction of the acidic proteins with the ribosome rather than the structural features responsible for their primary functions.

Amino Acid Sequence↗

[Directional velocity of the arterial flow, systemic vascular resistance and urinary excretion of sodium in cirrhotic patients].

OBJECTIVES: We have studied the vascular resistance at the posterior tibial artery utilizing the Doppler reverse/forward flow ratio, and its relationship to systemic vascular resistance and renal function in 32 nonazotemic cirrhotic patients. METHODS: Patients were divided into three groups. Group A comprised 10 patients without ascites or oedema; group B comprised 9 patients with ascites and a relatively high sodium excretion (40 +/- 34 mmol/day); and group C comprised 13 patients with ascites and very low sodium excretion (4.9 +/- 2 mmol/day). RESULTS: No significant differences were found in urine flow, creatinine or creatinine clearance between the three groups. Renin and aldosterone levels were found increased in group C. Systemic vascular resistance differed significantly in the three groups, being lower in group C. Significant higher values in the Doppler reverse/forward ratio were observed in patients with markedly increased sodium retention and less systemic vascular resistance (group C). The Doppler reverse/forward ratio showed significant correlations with systemic vascular resistance (r = 0.65; n = 32; p < 0.001), urinary sodium excretion (r = 0.53; n = 32; p < 0.01), renin (r = 0.474; n = 32; p < 0.01) and aldosterone levels (r = 0.589; n = 32; p < 0.001). CONCLUSIONS: These preliminary results suggest in patients with hepatic cirrhosis vascular resistance, assessed non-invasively, at the posterior tibial artery, increases with the severity of sodium retention and the impairment in systemic hemodynamics. Thus, this measurement may be useful for the evaluation and follow-up of patients with cirrhosis of the liver.

Adult↗

Simple strategy to induce antibodies of distinct specificity: application to the mapping of gp120 and inhibition of HIV-1 infectivity.

In this study 96 15-mer peptides encompassing the entire sequence of HIV-1 gp120 were synthesized and used to immunize BALB/c mice (i) alone or (ii) in conjunction with the T helper cell determinant FISEAIIHVLHSR (FIS) from sperm whale myoglobin, which is well recognized by major histocompatibility complex (MHC) class II molecules of BALB/c. Of these peptides 39 were immunogenic per se and 57 were not. Out of the 57 non-immunogenic peptides 53 could be rendered immunogenic with the second immunization protocol. With the exception of 4 cases, the anti-peptide antibody titers induced in (ii) were equal (14 cases) or higher (78 cases) than those induced in (i). From the 96 anti-peptide antibodies tested, 12 were able to recognize recombinant gp120 with good antibody titers, a result in agreement with previously identified B cell epitopes from gp120 by anti-peptide antibodies induced with longer peptides conjugated to a carrier protein. Moreover, 4 of the 12 anti-peptide antisera that recognized gp120 were able to neutralize HIV-1 infectivity in vitro, showing that the strategy of co-immunization with FIS may afford functional antibodies.

Amino Acid Sequence↗