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Biomedical subjects

G Esteban

Publications and source records attributed to G Esteban.

10 recordsLinked to original sources
PubMed2005

Lactic acid bacteria biomass monitoring in highly conductive media by permittivity measurements.

Recent technological improvements have extended the application range of permittivity biomass measurements to bacterial fermentations in highly conductive media. With Lactobacillus casei, the effective biomass detection sensitivity of the FOGALE Biomass System is around 0.2 g/l (0.01 pF/cm). Fermentations growth kinetics of Lactobacillus casei can be recorded with good reproducibility and accuracy despite the high medium conductivity varying between 15 and 75 mS/cm, and the low cell concentration (<6 g/l).

Biomass↗
PubMed2005

Dielectric monitoring of growth and sporulation of Bacillus thuringiensis.

On-line permittivity and optical density measurements have been used to monitor biomass concentration and sporulation status during growth of a spore-forming bacterium, Bacillus thuringiensis, in fed-batch culture. The correlation between permittivity, optical density and other observations showed three distinct phases of growth: growth itself, transition and sporulation. The permittivity variations during the transition and sporulation phases could be related to the sporulation development: the evolution pattern of the ratio of optical density to permittivity was representative of the culture state, and during the sporulation phase, a permittivity index could be build to measure the extend of spore liberation.

Absorption↗
PubMed1999

Cooling and freezing damage platelet membrane integrity.

Cytoskeletal rearrangements and a membrane lipid phase transition (liquid crystalline to gel) occur in platelets on cooling from 23 to 4 degrees C. A consequence of these structural alterations is irreversible cellular damage. We investigated whether platelet membrane integrity could be preserved by (a) previously studied combinations of a calcium chelator (EGTA) and microfilament stabilizer (cytochalasin B) with apparent benefit in protecting platelets from cooling injury or (b) agents of known benefit in protecting membranes and proteins from freezing injury. Platelet function and activation before and after freezing or cooling were measured by agglutination with ristocetin, aggregation with thrombin or ADP, platelet-induced clot retraction (PICR), and expression of P-selectin. Platelets were loaded with 10 nM fluorescein diacetate. After freezing or cooling, the preparations were centrifuged and the supernatant was measured for fluorescein. For cooling experiments, fresh platelets were chilled at 4 degrees C for 1 to 21 days with or without the combination of 80 microM EGTA/AM and 2 microM cytochalasin B (EGTA/AM-CytoB) and then warmed rapidly at 37 degrees C. For freezing experiments, 5% dimethyl sulfoxide (Me2SO) or 5 mM glycerol were added to fresh platelets. The preparations were then frozen at -1 degrees C/min to -70 degrees C and then thawed rapidly at 37 degrees C. Platelet membrane integrity, as measured by supernatant levels of fluorescein, correlated inversely with platelet function. Chilling platelets at 4 degrees C with EGTA/AM-CytoB showed a gradual loss of membrane integrity, with maximum loss reached on day 7. The loss of membrane integrity preceded complete loss of function as demonstrated by PICR. In contrast, platelets chilled without these agents had complete loss of membrane integrity and function after 1 day of storage. Freezing platelets in Me2SO resulted in far less release of fluorescein than did freezing with or without other cryoprotectants (P < 0.001). This result correlated with enhanced function as demonstrated by PICR and supports earlier observations that Me2SO protects platelet membranes from freezing injury. Release of fluorescein into the surrounding medium reflected loss of membrane integrity and function in both cooled and frozen platelets. Membrane cytoskeletal rearrangements are linked to membrane changes during storage. These results may be generally applicable to the study of platelet storage.

Adenosine Diphosphate↗
PubMed1999

Platelet membrane integrity during storage and activation.

BACKGROUND: The platelet cell membrane appears to undergo a lipid-phase transition on cooling from 23 degrees C to 4 degrees C. Consequences of this phase transition are leakage of cellular material and irreversible cellular damage. Whether agents, of known benefit in protecting membranes and proteins from cooling and drying injury, could also protect platelets was investigated. Leakage of cytosolic components was assessed by measuring the release of fluorescein into the surrounding medium. STUDY DESIGN AND METHODS: Fresh platelets were suspended in 5 percent dimethyl sulfoxide (DMSO) or in 5 mM of one the following agents: glucose, trehalose, sucrose, glycerol, ethylene glycol, 1,2-propanediol, or L-proline. Platelets were loaded with 10 nMfluorescein diacetate (FD), chilled at 4 degrees C for 24 hours or frozen at -1 degree C per minute to -70 degrees C, warmed rapidly at 37 degrees C, and centrifuged, and the supernatant was measured for the presence of fluorescein. The effect of FD on platelets was assessed by agglutination with ristocetin, aggregation with thrombin and ADP, platelet-induced clot retraction, and expression of p-selectin. Platelet function and activation before and after freezing or cooling were measured by the same methods. RESULTS: By flow cytometry, 98 percent of the platelets incorporated FD. The trapped fluorescein resulted in neither platelet activation (p = 0.9) nor reduction of platelet function (p = 0.12-0.94) from that in control platelets. Freezing of platelets in DMSO caused far less release of fluorescein than did freezing with other agents (p<0.001) or chilling of platelets at 4 degrees C for 24 hours (p<0.0001). Supernatant levels of fluorescein correlated inversely with platelet function. Fluorescein was also shown to be released during aggregation with thrombin or ADP but not during agglutination with ristocetin. CONCLUSIONS: Release of fluorescein into the surrounding medium indicated a loss of platelet membrane integrity and function. Cellular loading with FD is a simple method of studying membrane integrity of platelets and other cells.

Adenosine Diphosphate↗
PubMed1994

Some rumen ciliates have endosymbiotic methanogens.

Most of the small ciliate protozoa, including Dasytricha ruminantium and Entodinium spp. living in the rumen of sheep, were found to have intracellular bacteria. These bacteria were not present in digestive vacuoles. They showed characteristic coenzyme F420 autofluorescence and they were detected with a rhodamine-labelled Archaea-specific oligonucleotide probe. The measured volume percent of autofluorescing bacteria (1%) was close to the total volume of intracellular bacteria estimated from TEM stereology. Thus it is likely that all of the bacteria living in the cytoplasm of these ciliates were endosymbiotic methanogens, using H2 evolved by the host ciliate to form methane. Intracellular methanogens appear to be much more numerous than those attached to the external cell surface of ciliates.

Animals↗
PubMed1991

[Evaluation of 2 culture systems for the isolation of opportunistic mycobacteria].

The increment of infections produced by opportunist environmental mycobacteria (OEM) raises the question about the appropriateness of the conventional culture methods since they were initially designed to isolate M. tuberculosis. In this study we have comparatively evaluated the yield of a conventional culture (Löwenstein-Jensen) with respect to that of Middlebrook 7H12 (Bactec) in 396 selected samples in which OEM were isolated. Samples with commensal flora were previously subjected to a chemical homogenization-decontamination process with alkaline N-acetyl-cysteine. All cultures were sown with standard inoculates obtained from centrifugate material. The 7H12 culture showed a more rapid detection (mean: 17-18 days) and a greater effectiveness in the isolation of eventually pathogenic mycobacteria than the conventional Löwenstein-Jensen culture. The 7H12/LJ ratio for the species with high clinical relevance was: 1.3 for M. kansasii, 1.0 for M. marinum, 1.8 for M. scrofulaceum, 1.3 for M. xenopi, 1.5 for M. avium-intracellulare, 0.5 for M. chelonae, and 2.0 for M. fortuitum.

Bacteriological Techniques↗
PubMed1990

[Primary and initial bacterial resistances in tuberculosis patients at a general hospital, 1983-1987].

From july 1982 to december 1987 we prospectively evaluated the primary resistance (PR) of 248 strains of M. tuberculosis isolated from patients with several clinical forms of tuberculosis. The results were compared with those of a similar study carried out between 1978-1982. PR persisted relatively stable with time, although it showed a trend towards reduction. Isoniazide resistance (4.8%) still has a leading epidemiological interest. The 89% of strains showed isolated resistance to a single drug. Only two strains had associated resistance to INH and SM. Strains with PR to RIF, ETB or PZA were not detected. Of the 248 evaluated strains, 32 were isolated from intravenous drug abusers with AIDS. Only one of these strains had a low level (0.2 mg/l) PR to INH. At last two of these drug abusers developed acquired resistance to INH and ETB-RIF, respectively. Considering the difficulty to control chemotherapy in this group of patients, our finding has a considerable epidemiological interest.

Acquired Immunodeficiency Syndrome↗