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Biomedical subjects

M T García

Publications and source records attributed to M T García.

At least 37 records · Page 2Linked to original sources

Activity of voriconazole: post-antifungal effect, effects of low concentrations and of pretreatment on the susceptibility of Candida albicans to leucocytes.

This study examined: (i) the post-antifungal effect (PAFE) of Voriconazole (UK 109,496) on Candida albicans, at 2 concentrations (MIC and 4 x MIC) in the presence or absence of 10% human serum; (ii) the activity of low concentrations of the drug (1/4 x MIC) on yeasts that had previously been exposed to Voriconazole (PAFSE) with or without 10% human serum; and (iii) the effect of Voriconazole pretreatment on the fungicidal activity of leucocytes and serum against C. albicans (PALE). Two concentrations (0.25 and 1 mg/l) of Voriconazole induced no PAFE against C. albicans between -4.3 and -1.4 h, but when the assays were performed in the presence of serum, positive and concentration-dependent PAFEs were obtained (0.2-4.1 h). Pretreated yeasts were more susceptible than untreated yeasts to low concentrations (0.0625 mg/l) of Voriconazole, so the drug showed positive PAFSE that was dependent on the concentration used in pretreatment without serum (0.3-1.9 h) or with 10% human serum (0.5-2.5 h). Pretreatment of the growing C. albicans cells with Voriconazole (0.25 mg/l) increased their vulnerability to killing by leucocytes during the last 2 h (p < 0.05), leading to PALE of 2 h. The results suggest that these effects might be used to evaluate the in vivo activity of an antifungal agent. The sum of the durations of these effects (PAFE, PAFSES and PALE) cause a considerable delay in yeast growth in treated cultures compared with control cultures.

Antifungal Agents↗

Analysis of the genome of the gram-negative moderate halophiles Halomonas and Chromohalobacter by using pulsed-field gel electrophoresis.

The genomes of 11 moderately halophilic bacteria belonging to the genera Halomonas and Chromohalobacter have been analyzed by restriction endonuclease digestion and pulsed-field gel electrophoresis (PFGE). By using the infrequently cutting restriction endonucleases SpeI and SwaI, highly characteristic fingerprintings were obtained for each of the isolates studied. On the basis of the lengths of the SpeI and SwaI fragments, separated by PFGE, the genome size of the 11 strains studied was estimated. The genome size for 8 Halomonas strains ranged from 1450 to 2830 kb, whereas for the 3 Chromohalobacter strains studied it ranged from 1770 to 2295 kb. Finally, we show that macrorestriction fingerprints could be a useful tool to elucidate the taxonomic position of bacteria belonging to the Halomonas-Deleya complex.

DNA, Bacterial↗

[Recurrent infection by Streptococcus agalactiae].

BACKGROUND: To study the factors implicated in the infectious process (host, microorganism and antibiotic) of a newborn early sepsis by S. agalactiae that suffered a reactivation at day five from discharge. METHODS: Description of two episodes of newborn sepsis by S. agalactiae corresponding to the same patient and microbiologic study of the isolated strain: typing by "genomic macrorestriction" and antibiotic tolerance by "timed killing curves". RESULTS: It was demonstrated that both strains of S. agalactiae type la/c belonged to the same clone as well as the tolerance to ampicillin of the strain. DISCUSSION: This sort of infections processes in the newborn are very serious and there is possibility of relapse. Thus, it is important to study the ethiologic agent and its relationship with antibiotics, in order to stablish the best treatment regimes, avoiding the possibility of relapses as the case we have described.

Adult↗

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↗

Bone densitometry and biochemical bone remodeling markers in type 1 diabetes mellitus.

Bone mineral density (BMD) at the lumbar spine, quantified by dual energy X-ray absorptiometry, and biochemical bone remodeling markers (serum alkaline phosphatase, osteocalcin, tartrate-resistant acid phosphatase and urinary hydroxyproline) have been studied in 94 patients with diabetes mellitus aged 18-62 years. BMD was normal (1.13 +/- 0.02 g/cm2 in patients vs. 1.16 +/- 0.12 g/cm2 in controls), although it was found to be negatively correlated with HbA1, microalbuminuria, age and the duration of the disease. Serum alkaline phosphatase (188 +/- 75 I.U./l vs. 168 +/- 42 I.U./1; P < 0.03), serum tartrate-resistant acid phosphatase (14.3 +/- 4.3 I.U./l vs. 11.7 +/- 3.7 I.U./l; P < 0.0001) and urinary hydroxyproline (0.018 +/- 0.016 mmol/mmol creatinine vs. 0.011 +/- 0.008 mmol/mmol creatinine; P < 0.001) were higher in diabetics than in controls. Serum osteocalcin was lower (2.5 +/- 1.3 ng/ml vs. 3.4 +/- 1.2 ng/ml; P < 0.0001). No relationship was found between bone remodeling markers and BMD. It is concluded that lumbar BMD is normal in type 1 diabetic patients, although the degree of metabolic control, age and duration of the disease may affect it. In the light of the biochemical markers, bone remodeling may be disturbed in diabetes, but such disturbance seems to be unimportant regarding BMD.

Acid Phosphatase↗

Identification of Mycobacterium avium and Mycobacterium intracellulare isolated in Puerto Rico from clinical samples by the use of a non-radioactive DNA probe.

The Mycobacterium avium complex (MAC), especially M. avium, is an important opportunistic pathogen of AIDS patients in the United States. In Puerto Rico, the incidence of infections caused by MAC has not been determined. This is due, in part, to the difficulties associated to the microbiological identification of the microorganisms. In this work, a commercially available kit (AccuProbe, Gen-Probe, Inc., San Diego, CA) utilizing a DNA probe complementary to rRNA of M. avium and M. intracellulare was used to identify seventeen MAC strains and one unknown atypical mycobacterium recovered in culture in Puerto Rico from clinical samples. The results obtained revealed that M. avium was the predominant species recovered (83% of isolates tested). Only two cultures were identified as M. intracellulare. The unknown culture, which did not react with either probe, turned out to be M. gordonae. The probe tests not only are simple to perform, but provide cultural identification results in as little as two hours. This study, the first one of its kind in Puerto Rico, demonstrates that the nucleic acid probes for the cultural identification of M. avium and M. intracellulare offer the potential of providing a prompt diagnosis and much needed data on the epidemiology of MAC infections in Puerto Rico.

Bone Marrow↗

Hypersensitivity to latex, chestnut, and banana.

The incidence of latex-allergic patients is probably higher than suspected. A spectrum of IgE-dependent allergic reactions to latex products including urticaria, rhinitis, asthma, angioedema, and life-threatening anaphylaxis has been increasingly reported in recent years. We describe three patients with rubber hypersensitivity and allergy to fruit (banana and chestnut). Immediate positive responses were obtained in prick tests with latex, banana, and chestnut extracts. Histamine release was positive and specific IgE antibodies to all three extracts were detected by fluorescence radioimmunoassay. In the RAST-inhibition studies, the extract of latex inhibited the binding of chestnut and banana, but chestnut and banana extracts did not inhibit the binding of latex. These results suggest a sensitivity to crossreacting antigens in latex allergy associated with allergy to certain fruits.

Adult↗

A series of annexins from human placenta and their characterization by use of an endogenous phospholipase A2.

Membranes from human placenta contain proteins which inhibit the activity of phospholipases A2 by binding to phospholipid thus impeding substrate availability. We used unilamellar mixed liposomes and a partially purified cytosolic phospholipase A2 from placenta for characterizing this substrate-depleting activity. A major portion of these inhibitory proteins was released by extracting washed membranes with a Ca+(+)-chelator. Biochemical fractionation and systematic analysis resulted in the unequivocal identification of a series of annexin proteins. We describe a straightforward procedure which allows to obtain 8 annexins from placenta either in pure form or as a mixture of two annexins. One of them was obtained in two forms which had the same molecular mass of 68 kDa but differed in charge. We also present suggestive evidence for a novel annexin I-related polypeptide of Mr 45,000 which is an excellent in vitro substrate for protein kinase C. We estimate that about 2% of the total placental membrane proteins are annexins. For achieving half inhibition of phospholipase A2 activity with pure annexins, up to a 6.5-fold difference in the amounts of protein was observed when calculated on a molar basis. This suggests specificity of individual annexin species.

Annexins↗

The C57BL/10Bg sps/sps mouse: a mutant with absence-like seizures; neurochemical and behavioral correlates.

C57BL/10Bg sps/sps mice display behavioral arrest, similar to generalized absence seizures. Compared with the parent strain C57BL/10Bg SPS/SPS, the activities of glutamate decarboxylase (GAD, E. C. 2.6.1.15), GABA aminotransferase (GABA-T, E. C. 2.6.1.19), aspartate aminotransferase (ASP-T, E. C. 2.6.1.1), and glutamate dehydrogenase (GDH, E. C. 1.4.1.3) in whole brain crude supernatant were significantly reduced in the sps/sps mice. Alanine aminotransferase activity (ALA-T, E. C. 2.6.1.2), was not altered in any of the strains, and normalization of GAD, GABA-T and GDH activities by that of ALA-T, further revealed significant differences between the normal strain (SPS/SPS), the heterozygotes (SPS/sps), and behavioral arrest (sps/sps) mice. These results suggest the possible involvement of GABAergic and glutamatergic neurotransmission in the absence-like behavior displayed by sps/sps mice. Open field behavior of C57BL/10Bg sps/sps mice is characterized by periods of marked inactivity which easily distinguish affected homozygotes, from their heterozygotes littermates.

4-Aminobutyrate Transaminase↗

Relationships among Vibrio costicola strains assessed by DNA-DNA hybridization.

Deoxyribonucleic acid (DNA)-DNA hybridization was used to determine the relationships among 14 strains of Vibrio costicola isolated from cured meats or hypersaline waters. Our results indicate that the strains investigated should be considered members of the same genomic species, with homologies of 60% or higher with the V. costicola strains used as reference. Low DNA relatedness was found with other marine vibrios or moderately halophilic bacteria tested. The guanine-plus-cytosine base contents of DNAs from the 14 V. costicola strains were 47.6 to 51.1 mol%.

Base Composition↗

Influence of temperature and concentration on the postantifungal effect and the effects of sub-MIC concentrations of four antifungal agents on previously treated Candida species.

BACKGROUND: The objective of this study was to investigate the impact of different temperatures (22, 35 and 37 degrees C) on the postantifungal effect (PAFE) and the effect of sub-MIC concentrations (1/4 x MIC) on Candida albicans and Candida glabrata in PAFE stage (PAFSE). METHODS: This stage was induced by a 1.5-hour pretreatment with different doses (1 x, 4 x and 8 x MIC) of four antifungal agents that are fundamental to modern candidiasis therapy. RESULTS: The temperature, as well as the dose of the antifungal agent that was applied during the pretreatment, determined the duration of the two studied effects. An increase in the temperature and/or the dose prolonged the duration of the PAFE and PAFSE in both species, independent of the applied antifungal agent. Amphotericin B and 5-fluorocytosine always induced significant PAFEs (0.5-4.8 h and 0.5-3.0 h, respectively), which were increased (0.7-3.4 h and 0.5-3. 2 h, respectively) by posterior exposure to 1/4 x MIC of the respective antifungal agent. In the case of ketoconazole and fluconazole, temperature and concentration were especially important. Although neither antimycotics was able to induce a significant PAFE, posterior exposure to 1/4 x MIC of each of the two azoles led in both yeast species to a significant PAFSE of up to 0.8 h (if the concentrations and/or the temperatures were high enough). CONCLUSION: Factors such as temperature and concentration could be important when choosing an antifungal agent.

Antifungal Agents↗

Influence of human serum on the postantifungal effect of four antifungal agents on Candida albicans.

This study evaluates the influence of both fresh and heated human serum on the postantifungal effect (PAFE) induced by different concentrations of amphotericin B (AmB), 5-fluorocytosine (5-Fc), Ketoconazole (Kz) and fluconazole (Flu) on two strains of Candida albicans. The concentrations were selected in harmony with the pharmacokinetic properties and toxicity of the drugs. Without serum there was no delay in the growth of yeast cultures pretreated with Kz or Flu, leading to negative PAFEs, however with AmB and 5-Fc the PAFEs were positive. When assays were made in the presence of 10% fresh human serum, the duration of the PAFEs increased with all drugs tested, and those induced by azolic agents became positive. In the presence of 10% human serum heated at 56 degrees C for 30 min, the PAFEs of the antifungal agents were similar to those obtained in the absence of serum. Our results suggest that fresh serum positively influenced PAFE which may be an important factor in determining the dosing regimen for infection by yeasts.

Amphotericin B↗