Search PubMed⌕ Search

Biomedical subjects

J Fernandez

Publications and source records attributed to J Fernandez.

At least 91 records · Page 5Linked to original sources

A lipoamide dehydrogenase from Neisseria meningitidis has a lipoyl domain.

A protein of molecular weight of 64 kDa (p64k) found in the outer membrane of Neisseria meningitidis shows a high degree of homology with both the lipoyl domain of the acetyltransferase and the entire sequence of the lipoamide dehydrogenase, the E2 and E3 components of the dehydrogenase multienzyme complexes, respectively. The alignment of the p64k with lipoyl domains and lipoamide dehydrogenases from different species is presented. The possible implications of this protein in binding protein-dependent transport are discussed. This is the first lipoamide dehydrogenase reported to have a lipoyl domain.

Amino Acid Sequence↗

Formation of the female pronucleus and reorganization and disassembly of the first interphase cytoskeleton in the egg of the glossiphoniid leech Theromyzon rude.

Eggs of the leech Theromyzon rude were studied under dissecting, fluorescence, light, and electron microscopes. The sperm-derived centrosome divides and separates at early first interphase. The female pronucleus forms beside the male pronucleus at the centrally located perinuclear plasm domain, by fusion and remodelling of karyomeres that have descended from the animal pole at the tip of an ooplasmic process (centripetal ooplasmic flow). Retraction of this process (centrifugal ooplasmic flow) appears to return ooplasm to the animal pole. The reversible flow of ooplasm occurs along a subset of monaster microtubules that exhibit delayed depolymerization, and organelles thus move in both directions. Depolymerization of the first interphase monaster fibers seems to occur as a regular wave, moving from the internally located perinuclear plasm to the peripherally situated egg poles. However, disassembly of the ectoplasmic monaster fibers is preceded by furrowing and reorganization of both microtubules and microfilaments along polar rings and meridional bands. Bipolar contraction of microfilaments and shortening of microtubules lead to organelle concentration at the egg poles.

Actins↗

Comparison of low-pressure versus standard-pressure fixation Carpentier-Edwards bioprosthesis.

Intermediate-phase clinical results of 51 low-pressure (LP) and 234 standard-pressure (SP) fixation porcine Carpentier-Edwards (CE) valves implanted between 1977 and 1991 were compared for valve-related events. Group similarities included New York Heart Association functional class, ejection fraction, and sex. Patients with SP valves were younger (mean age, 58 versus 68 years; p = 0.0001). There were 20 in-hospital deaths (8.6%) in the SP valve group and 5 (9.8%) in the LP valve group (p = 0.79). Follow-up was 99%, with a mean of 104 months in the SP valve group versus 55 months in the SP valve group (p = 0.0001). The actuarial survival rate was 48.2% and 22.3% at 10 and 15 years, respectively, in the SP valve group and 34.1% at 10 years in the LP valve group (p = 0.42). Freedom from events at 5, 10, and 15 years in the SP valve group and at 5 years in the LP valve group was as follows: for late valve-related events, 86.3%, 51.4% and 20.2%, respectively, in the SP valve group versus 85% in the LP valve group (p = 0.44); for valve-related death, 96.4%, 93.6%, and 87.3% in the SP valve group versus 100% in the LP valve group (p = 0.20); for structural valve failure, 96%, 68%, and 35% in the SP valve group versus 100% in the LP valve group (p = 0.09); and for reoperation, 95%, 61%, and 30% in the SP valve group versus 92% in the LP valve group (p = 0.82). In conclusion, this study revealed no significant statistical difference between LP and SP valves. In the LP valve group, structural valve failure/valve-related death was not observed, perhaps indicating a more favorable result. Absolute verification of this trend awaits long-term follow-up.

Actuarial Analysis↗

CABG in octogenarians: early and late events and actuarial survival in comparison with a matched population.

BACKGROUND: With important demographic changes in cardiac surgical practice, more older patients are undergoing complex cardiac operations. Controversy exists as to whether the expenditure of healthcare resources on the growing elderly populations represents an effective approach in maintaining a meaningful quality of life. METHODS: From January 1982 through April 1991, 121 consecutive octogenarians underwent a surgical procedure that included coronary artery bypass grafting. Retrospective review of patient medical records was performed; follow-up information was obtained via telephone contact with the patient, the patient's family, or the patient's physician. RESULTS: There were 67 men (55%) and 54 women (45%). Mean age was 82.1 years (range, 80 to 89 years). Sixty-nine percent of the patients were having class III or IV symptoms. There were 11 hospital deaths (9.1%); risk factors included longer cardiopulmonary bypass time (p = 0.01), higher preoperative left ventricular end-diastolic pressure (p = 0.02), advanced age (p = 0.05), history of renal disease (p = 0.02), and myocardial infarction (p = 0.04). Late death occurred in 34 patients (30.9%) at a mean of 27 months postoperatively; univariate risk factors included chronic obstructive pulmonary disease (p = 0.009), higher left-ventricular end-diastolic pressure (p = 0.03), and recent myocardial infarction (p = 0.03). Actuarial survival, including hospital death, was 32.8% at 80 months, compared with 37.6% for an age; sex; and race-matched population (p > 0.3). Most late survivors (84%) were in New York Heart Association class I or II. CONCLUSIONS: We conclude that coronary artery bypass grafting can be performed in octogenarians with an acceptable, although increased risk. Hospital survivors have a good late functional status but are at risk for pulmonary and other atherosclerosis-related events, which impair overall survival.

Aged↗

Perioperative events in patients with failed mechanical and bioprosthetic valves.

From 1963 through 1991, 1037 patients underwent reoperative valvular procedures. The 478 patients having reoperations for either failed bioprosthetic (n = 212) or mechanical (n = 266) valves were evaluated. There were 210 male (44%) and 268 female (56%) patients. The mean age at reoperation of the patients in the bioprosthesis group was 59.7 years and and that in the mechanical valve group was 56.1 years (p = 0.0006). The mean interval to the time of reoperation was 84.7 months in the mechanical valve group and 74 months in the bioprosthesis group. There was no difference between the two groups in the functional class at reoperation. More severe mitral valve stenosis and incompetence, more severe aortic valve stenosis, and higher right ventricular and pulmonary arterial pressures were noted in the bioprosthesis group than in the mechanical valve group. Hemolysis (p = 0.05) was more prevalent in the patients with mechanical valves than in the ones with bioprostheses. A longer aortic occlusion time (p = 0.0001) and longer cardiopulmonary bypass time (p = 0.0001) were required for the reoperations in the bioprosthesis group. The operative mortality was 13.2% for the bioprosthesis patients and 12.4% for the mechanical valve patients. The risk factors for hospital death included the cross-clamp time (p = 0.0001), the functional class (p = 0.00001), the presence of ascites (p = 0.02), hepatomegaly (p = 0.002), and decreasing ejection fraction (p = 0.05). We conclude that mechanical valve failures do not produce catastrophic events resulting in poor reoperative results.

Adolescent↗

Purification and characterization of Acinetobacter calcoaceticus 4-hydroxybenzoate 3-hydroxylase after its overexpression in Escherichia coli.

4-Hydroxybenzoate 3-hydroxylase [EC 1.14.13.2] from Acinetobacter calcoaceticus was purified to homogeneity following the 40-fold overexpression of this gene (pobA) in Escherichia coli. Overexpression was accomplished by placing the folA gene (encoding trimethoprim-resistant dihydrofolate reductase) directly downstream of the pobA gene, and demanding growth of recombinants on elevated concentration of trimethoprim. Presumably, the surviving variants have undergone a genetic alteration which allowed the overexpression of both folA and pobA. 4-Hydroxybenzoate 3-hydroxylase was purified in two chromatographic steps, characterized biochemically, and its properties were compared to those of its homolog from Pseudomonas fluorescens. The two enzymes differ in their response to Cl- ion inhibition. A single amino acid change in the putative NADPH-binding site is proposed to account for this difference. The inhibitory and catalytic properties of substrate analogs were also examined.

4-Hydroxybenzoate-3-Monooxygenase↗

Activation of cytoplasmic trehalase by cyclic-AMP-dependent and cyclic-AMP-independent signalling pathways in the yeast Candida utilis.

Derepressed cells of Candida utilis suspended in buffer exhibited both a transient cAMP-mediated signal and a marked activation of cytoplasmic trehalase when supplemented with glucose. Nitrogen sources or protein synthesis inhibitors, as well as protonophores or uncouplers, were also able to cause trehalase stimulation in derepressed cells even in the absence of the sugar. The increase in trehalase activity caused by nitrogen sources or protein synthesis inhibitors was not accompanied by changes in cAMP levels. Moreover, acridine orange inhibited both the cAMP signal and the glucose-induced activation of trehalase without affecting the increase in trehalase activity caused by nitrogen sources or protein synthesis inhibitors. These results suggest that cAMP is not involved as second messenger in the signal for trehalase stimulation induced by the latter compounds. By contrast, the addition of glucose to repressed cells suspended in buffer failed to cause the cAMP-mediated glucose signal and sugar-induced trehalase activation. No significant changes in either trehalase activity or cAMP concentration were observed upon addition to these cells of asparagine, cycloheximide, anisomycin or other agents, including protonophores and uncouplers. However, heat treatment of repressed cultures resulted in a moderate increase in trehalase activity with negligible change in cAMP levels, whereas such an effect was not observed in derepressed cultures. The thermally induced increase in trehalase activity was dependent on de novo protein synthesis and required the presence of glucose.(ABSTRACT TRUNCATED AT 250 WORDS)

Candida↗

Comparative study between European and American species of Polistes using sera from European sensitized subjects.

Although the different vespid species of the genus Polistes are found all over the world, studies about allergy sensitization to these species have been made mainly in North America. We studied the specific IgE in a large group of sera from patients sensitized to Polistes dominulus (PD) and determined the crossreactivity with another important European Polistes (P. gallicus [PG]) and with five American relevant species of this genus: P. exclamans (PE), P. fuscatus (PF), P. metricus (PM), P. annularis (PAN) and P. apachus (PAP). Studies indicated that 96.7% of sera with IgE positive to PD were positive to PG. No case positive to just PG was found. The comparison of these two European species with the American relatives indicated that most cases were positive to all the insects although there were significant differences in the radioallergosorbent test (RAST) value. These results were confirmed by RAST inhibition studies, and indicate that although the European and American species are closely related, species specific allergenic differences exist. These data suggest that in vivo studies should be carried out in order to determine the sensitivity and specificity of the European venoms compared with the American species.

Animals↗

Nonimmediate reactions to betalactams: prevalence and role of the different penicillins.

In patients treated with penicillins, adverse cutaneous reactions can occur within minutes or may take several days to develop. IgE antibody-mediated reactions are well documented, but other mechanisms may also be involved. In particular, nonimmediate reactions have not been studied extensively, and the purpose of the present work was to establish the incidence of such reactions among a large group of patients and to study the penicillins involved. A total of 380 subjects with a history of a cutaneous reaction following administration of a penicillin antibiotic was included in the study. Skin tests and specific IgE measurements (RAST) were carried out using various penicillins and penicillin-related reagents, and patients were also challenged with various penicillins. In some patients with delayed skin test responses, skin biopsies were carried out. The tests confirmed that 74 subjects (19.4% of total investigated) had suffered a cutaneous reaction to a penicillin derivative, and 29 of these subjects (7.6% of total or 39% of confirmed) showed evidence of having suffered a nonimmediate reaction. The latter group were identified by giving a positive delayed direct challenge, and in 65% of the cases a delayed skin test response was detected. In most cases, these responses were to amino penicillins. Skin biopsies showed a lymphomonocytic cell infiltrate. Nonimmediate reactions to penicillins are a reproducible phenomenon, suggesting that a specific mechanism is responsible. By direct challenge, 93% of responders were positive to amino penicillins (10.3% ampicillin, 82.7% amoxicillin), indicating a major role for these penicillins in nonimmediate reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

[Efficacy and tolerance of praziquantel (Biltricide) in the treatment of distomatosis caused by Fasciola hepatica].

OBJECTIVES: Due to the side-effects of dehydroemetine, we have chosen praziquantel, a broad-spectrum antihelmintic, as a treatment for distomatosis secondary to Fasciola hepatica in humans. The aim of this retrospective study was to evaluate the efficacy and tolerance to praziquantel in patients with this disease. METHODS: Twenty-five patients (12 men) with a definite diagnosis of distomatosis and no previous treatment were followed-up between 8 months and 3 years (> 18 months in 76% of cases). The follow-up was based on clinical, biochemical and serological criteria. All patients received praziquantel (75 mg/kg/day orally) for 5 days. Treatment was started after endoscopic or surgical removal of parasites locolized in the biliary tract, in two patients. A similar therapeutic course was administered twice in four patients with persistent clinical symptoms, hypereosinophilia or arch 2 on immunoelectrophoresis. RESULTS: Cumulative rates of patients with normalized eosinophilia and seronegativation at 6, 9 and 12 months were 55, 65, 75% and 55, 70, 100%, respectively. Complete recovery occurred in 18 patients (72%) whereas hypereosinophilia persisted for more than one year in 5 patients. No side-effects, except transient nausea in a few cases, were observed. CONCLUSION: Since praziquantel seems to be both effective and well tolerated in a large proportion of patients, this drug can be recommended as a first choice for distomatosis due to Fasciola hepatica in human.

Adult↗

Normothermic retrograde cardioplegia is effective in patients with left ventricular hypertrophy. A prospective and randomized study.

Twenty patients with left ventricular hypertrophy (LVH) undergoing isolated aortic valve replacement were prospectively randomized to receive either continuous retrograde normothermic (n = 8) or intermittent retrograde hypothermic (n = 12) methods of myocardial protection. Biopsies of the left ventricular septum were evaluated for ultrastructure and assayed for ATP. There was no mortality, no requirement for intra-aortic balloon pump nor neurological events in any of the patients from either group. Myocardial ATP (warm 23.2 +/- 1.8 nmol/mg protein; cold 22.4 +/- 1.2 nmol/mg protein; p = 0.72) and myocardial CPK-MB (warm 43.6 +/- 5.2 U/l; cold 39.0 +/- 2.5 U/l; p = 0.67) were not significantly different. Ultrastructure was generally well preserved in the biopsies from both groups, with the exception of one patient in the normothermic group. Systemic lactate sampled after 40 minutes of cardiopulmonary bypass was significantly higher in the normothermic group (warm 3.4 +/- 0.27 mmol/l; cold 2.3 +/- 0.21 mmol/l; p = 0.01), however, the myocardial lactate production was not significantly different between the two groups (extraction ratio; warm 0.01 +/- 0.3; cold 0.13 +/- 0.1; p = 0.45). We conclude that the continuous normothermic retrograde method of myocardial protection is effective in patients with left ventricular hypertrophy; however, the higher systemic lactate levels using this technique raises concerns regarding the adequacy of systemic perfusion at 37 degrees C.

Aged↗

A two-year study on the efficacy and tolerability of policosanol in patients with type II hyperlipoproteinaemia.

This is a report of the results of a two years' randomized, double-blind placebo-controlled study of the efficacy, safety, and tolerability of policosanol administered at 5 mg twice-a-day in the treatment of type II hyperlipoproteinaemia. The study included 69 patients from both sexes, in whom total cholesterol and low-density-lipoprotein cholesterol (LDL-C) were not controlled sufficiently by diet. The treatment effect on total cholesterol and LDL-C was maintained during the 2-year follow up. Thus, percent reductions 24 months after therapy were 25% (LDL-C) and 18% (cholesterol). All comparisons with placebo were significant. Similarly, ratios of LDL-C to HDL-C and cholesterol to HDL-C were significantly reduced and such decreases were maintained during the study. Policosanol raised significantly the values of high-density lipoprotein cholesterol (HDL-C) during the study and maximal increases were reached 12 months after therapy (+21%). From this time the increases mildly declined to +14% and +11.2% respectively at 18 and 24 months after therapy. No significant changes in triglycerides were observed as compared with baseline or placebo. No patient withdrew from the study because of adverse effects. No drug-related clinical or biochemical adverse side-effects were observed. Any adverse experiences reported were mild and transient; moreover, no significant differences were obtained when compared with those reported by the placebo group. The results indicate that policosanol administered for two years to patients with type II hypercholesterolaemia shows a maintained efficacy as well as very good safety and tolerability.

Anticholesteremic Agents↗

Localization of antigenic sites on human cytomegalovirus virion structural proteins encoded by UL48 and UL56.

Two human cytomegalovirus (HCMV) virion structural proteins and their associated reading frames have been identified with two human-derived monoclonal antibodies (HMAbs), X2-16 and X-16. HMAb X2-16 identified recombinant protein expressing molecular clones that mapped to the open reading frame (ORF) of the UL48 gene of HCMV, between amino acids 584 and 646 (nucleotides 65,084 and 65,272, Chee et al., 1990, "Current Topics in Microbiology and Immunology," Vol. 154, pp. 125-169). The UL48 gene product has an apparent molecular weight of 216 kDa. HMAb X-16 identified clones derived from the UL56 ORF between amino acids 380 and 425 (nucleotides 84,733 and 84,870). On immunoblots, HMAb X-16 detected two HCMV proteins of 96 and 60 kDa. Both UL48 and UL56 are highly conserved among the human herpesviruses and their products have been predicted to have essential functions for virus production and maturation. These results confirm that UL48 and UL56 are functional genes encoding essential viral proteins which also generate an immune response in the immunocompetent host.

Amino Acid Sequence↗

An improved procedure for enzymatic digestion of polyvinylidene difluoride-bound proteins for internal sequence analysis.

An improved and simplified procedure for enzymatic digestion of proteins bound to polyvinylidene difluoride (PVDF) membranes for obtaining internal protein sequence data is presented. This improved procedure is compatible with various enzymes (trypsin, endoproteinase Lys-C, endoproteinase Glu-C, and clostripain) and is performed in the presence of 1% hydrogenated Triton X-100 (RTX-100)/10% acetonitrile/100 mM Tris, pH 8.0, followed by microbore HPLC purification of the recovered peptides. Previously published techniques required treatment of the PVDF-bound protein with polyvinylpyrrolidine M(r) 40,000 (PVP-40) prior to digestion, in order to prevent adsorption of the enzyme to the membrane. Unfortunately, contaminants produced from residual PVP-40 interfere with subsequent peptide mapping. We have found that when RTX-100 is used in the digestion buffer, no pretreatment of the PVDF-bound protein with PVP-40 is necessary. Advantages of this improved (one-step) procedure over the two-step PVP-40 procedure are (a) the elimination of undesirable contaminants associated with PVP-40, (b) a decrease in the time required for the technique, and (c) a reduction in sample manipulation. Peptide maps and recoveries from PVDF-bound standard proteins (4 micrograms each) enzymatically digested with this one-step method are compared with those obtained from the standard PVP-40 method. In addition, peptide maps and internal sequence data from low-level quantities of unknown proteins enzymatically digested with the improved procedure are presented. To date, this improved, one-step procedure has been successfully applied to 52 PVDF-bound unknown proteins (0.7-10 micrograms) of varying molecular weight (19-300 kDa) for which internal sequence data were obtained.

Amino Acid Sequence↗