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Biomedical subjects

A Pulido

Publications and source records attributed to A Pulido.

14 recordsLinked to original sources
PubMed2006

Influence of Fe concentration in the medium on multicellular pollen grains and haploid plants induced by mannitol pretreatment in barley (Hordeum vulgare L.).

This study aims to clarify the short- and long-term effects of the iron concentration in the medium on androgenesis induced in barley by isolated microspore culture. The ultrastructural features and pectin composition of the intine wall were studied in the initial stages of androgenesis. The evolution of electron-dense iron deposits on the intine was analysed in multicellular pollen grains obtained by isolated microspore culture performed for 3, 6, and 9 days using various concentrations of FeNa(2) EDTA. Finally, the number of embryo-like structures and green plants obtained by microspore culture using different Fe concentrations was evaluated in order to estimate the optimum concentration for isolated microspore culture.

Biological Evolution↗
PubMed2005

Cytological and ultrastructural changes induced in anther and isolated-microspore cultures in barley: Fe deposits in isolated-microspore cultures.

To gain further insight into the role played by sporophytic anther tissues in the early stages of the androgenic process, we have compared the cytology and ultrastructure of barley embryogenic pollen grains obtained by anther culture with those obtained by isolated-microspore culture. The microspores behaved similarly in both culture systems but ultrastructural studies detected a significant difference: the presence of electron-dense deposits on the intine of embryogenic pollen grains generated by isolated-microspore culture compared to their absence in grains generated by anther culture. To discover the nature of these deposits, we applied proteinase K and EDTA treatments to ultrathin sections. We also subjected the deposits to X-ray microanalysis and found that they contained iron. Anthers and isolated microspores were cultured in media containing different concentrations of iron so as to evaluate the presence of these deposits on the intine. Deposits were not found in anther cultures at any iron concentration used or in microspore cultures when concentrations were lower than 40 mg/L. The Fe deposits on the intine appear to derive from an excess of Fe in the isolated-microspore culture medium which, if allowed to pass through the cell wall, could well be toxic to the embryogenic development of the microspores.

Cells, Cultured↗
PubMed2000

[Evaluation of four rapid methods for the investigation of the toxigenic capacity of Clostridium difficile strains isolated in a selective medium].

BACKGROUND: Use of selective Clostridium difficile culture as a diagnostic method for C. difficile associated disease requires to prove the toxigenic ability of the isolates. Toxin B detection by cell culture assay after growing the microorganism in enriched broth is the standard method, but it delays the final diagnosis for 3-5 days. This study compares retrospectively four rapid techniques for detecting these toxigenic C. difficile strains. METHODS: 106 clinical isolates of C. difficile (72 toxigenic and 34 non-toxigenic), these and 16 clinical strains of other species of Clostridium were investigated. The four methods were performed directly from colonies growing on solid agar. They were: a) cytotoxin detection in cell culture; b) two PCR amplifications of toxin A and toxin B, respectively, and c) toxin A detection by an immunoenzymatic method (VIDAS CDA2). All these procedures were completed within a normal working day. RESULTS: Only the 72 toxigenic C. difficile strains gave positive results by cell culture and PCR techniques (sensitivity and specificity: 100%). A total of 14 out of 49 toxigenic C. difficile strains showed negative results by the VIDAS assay in the first run, but all them were positive in repeated tests. CONCLUSIONS: Although all methods performed well, the cytotoxicity assay done directly on colonies growing in CCFA is a simple and rapid technique, and appears to be well-suited for use in laboratories with access to cell cultures.

Agar↗
PubMed1997

Sepsis due to group G Streptococcus after a total hip arthroplasty. A case report.

A patient with a total hip arthroplasty developed an aggressive infection with group G Streptococcus. Very few similar cases have been reported, but they all resolved with antibiotics or drainage. A Girdlestone resection was necessary in our case because of loosening and extensive bony destruction. The true incidence may be greater than that reported and the prognosis may be worse.

Aged↗
PubMed1996

Molecular tracking of coagulase-negative staphylococcal isolates from catheter-related infections.

Three molecular typing methods (pulsed-field electrophoresis, localization of the mecA gene, and probing the vicinity of mec) have been used for the characterization of 40 catheter-related isolates of coagulase-negative staphylococci (CNS) in 14 patients admitted to the same hospital. The 40 isolates yielded 14 different SmaI banding patterns and corresponding unique localizations of mecA, each associated with a unique ClaI mecA polymorph. In 6 of the 14 patients the contaminated skin at the catheter entry site was the source of 4 local infections and 2 cases of bacteremia. A contaminated hub was the origin of 2 local infections and 4 cases of bacteremia in 6 more patients. The remaining 2 patients had positive cultures from both skin and catheter hub. In each bacteremic patient, the CNS recovered from catheter-related sites (tip, skin, and/or hub) and the CNS recovered from blood were identical, but each of these matching isolates was unique to the particular patients, indicating a low rate of cross-infection from patient to patient. Although classical methods for typing CNS (e.g., biotype and antibiotype) are readily available for most hospital laboratories, they have limitations concerning reproducibility and discriminatory power. Molecular epidemiologic techniques can provide powerful support to traditional techniques in determining the etiologic role of CNS in the disease process.

Bacterial Proteins↗
PubMed1991

Progesterone metabolism during storage of blood samples from Gyr cattle: effects of anticoagulant, time and temperature of incubation.

Ten Gyr cows with a functional corpus luteum were used to evaluate the effects of time and temperature of incubation of blood samples on progesterone (P4) concentrations detected in plasma or serum. From each cow, a blood sample was collected into a flask containing no anticoagulant, another into an heparinized flask and a third into a flask containing sodium fluoride. The blood from each flask was divided into 46 aliquots. One of them was centrifuged within 5 min of collection. The remaining 45 aliquots were divided into three groups and kept at three different temperatures: 4 degrees C, 17 degrees C, or 37 degrees C. For each anticoagulant, aliquots from every cow and incubation temperature were centrifuged every 30 min for 6 h, and then at 8, 12 and 24 h. Plasma or serum were separated immediately after centrifugation and were kept frozen at -20 degrees C until assayed for progesterone. The mean initial concentration of P4 in serum (8.3 ng/ml) significantly diminished (P<0.05) to 6.7 ng/ml after 5 h of incubation at 4 degrees C, 3 h at 17 degrees C, or 2 h at 37 degrees C. In plasma from heparinized blood the initial concentration (7.8 ng/ml) declined significantly after 6 h of incubation at 4 degrees C, 2 h at 17 degrees C, or 1 h at 37 degrees C. Sodium fluoride used as anticoagulant prevented the degradation of P4 since the initial concentration of P4 (6.7 ng/ml) never declined during incubation at either 4 degrees C or 37 degrees C; the only significant reduction occurred after 24 h of incubation at 17 degrees C.

Journal Article↗
PubMed1991

[Multicenter study of the Rosco-Neisseria system for the identification of pathogenic neisserias and Branhamella catarrhalis].

The commercial Rosco-Neisseria system was evaluated in the identification of 228 oxidase-positive Gram-negative diplococci and it was compared with conventional tests. The procedure detects gamma-glutamyl aminopeptidase, ONPG, tributyrin hydrolysis, and sensitivity to the disk of 10 micrograms of colistin. A correct identification was obtained in the 65 strains of Neisseria gonorrhoeae, the 33 of N. meningitidis, the 12 of N. lactamica, and the 56 of B. catarrhalis. The method was also able to discriminate 54 out of the 62 strains of nonpathogenic Neisseria. However, the 7 strains of Neisseria polysaccharea and one strain of N. subflava biovar perflava were erroneously identified as N. gonorrhoeae. None of the latter was superoxol positive in contrast with the 100% of cases of gonococcal strains. The Rosco-Neisseria system is simple and inexpensive but it should be applied on specimens that grow on selective media for gonococci (such as Thayer-Martin and others) and it should be complemented by superoxol test.

Bacterial Infections↗
PubMed1990

Treatment of pilonidal sinus by excision and rhomboid flap.

A series of 23 patients with chronic pilonidal disease have been treated by excision and transposition rhomboid flap. Full primary healing was obtained in all patients, with only two cases of wound seroma. The average hospital stay was 9 days. The mean follow-up period was 12 months, and no late recurrences have occurred.

Adolescent↗
PubMed1990

Superoxol and aminopeptidase tests for identification of pathogenic Neisseria species and Moraxella (Branhamella) catarrhalis.

The superoxol test, and prolyl aminopeptidase and gammaglutamyl aminopeptidase tests were evaluated for the detection of pathogenic Neisseria spp. using 317 strains of Neisseria-ceae. The superoxol test was positive for all 116 gonococci and 62 Moraxella (Branhamella) catarrhalis strains, but also for three strains of Neisseria meningitidis, one strain of Neisseria lactamica and eight saprophytic neisseriae. When using strains grown on Thayer-Martin medium, the positive and negative predictive values of the superoxol test for the identification of Neisseria gonorrhoeae were 96.7% and 100% respectively. Meningococci were the only neisseriae growing on Thayer-Martin medium that showed gamma-glutamyl aminopeptidase activity. The prolyl aminopeptidase test showed low specificity.

Aminopeptidases↗
PubMed1990

Butyrate esterase (4-methylumbelliferyl butyrate) spot test, a simple method for immediate identification of Moraxella (Branhamella) catarrhalis [corrected].

A total of 47 Moraxella (Branhamella) catarrhalis strains, 89 strains of Neisseria spp., and 82 strains of miscellaneous bacteria and yeasts were studied by using a fluorogenic spot method which detects butyrate esterase. A positive butyrate esterase spot test correctly differentiated M. catarrhalis from Neisseria spp., which had a negative butyrate esterase spot test reaction. The test is rapid, simple, and easy to perform. The butyrate esterase spot test was useful for direct identification of M. catarrhalis from primary cultures. However, false-positive reactions may occur with mixed cultures.

Bacteriological Techniques↗
PubMedDate not supplied

Use of a commercial double-test tablet (Rosco PGUA/indole) for screening of Escherichia coli.

A commercial double-test tablet (Rosco PGUA/indole) for detection of beta-glucuronidase (beta-GUR) activity and indole production was evaluated on a collection of 393 isolates of Enterobacteria. Both beta-GUR and indole were positive on 96.6% of Escherichia coli strains. beta-GUR, only, was also detected in 25 Shigella spp., four Enterobacter cloacae, eight Citrobacter freundii, and five Salmonella enteritidis strains, none of which were indole producers. An additional 261 consecutive clinical isolates of oxidase-negative nonswarming Gram-negative bacilli were studied in a parallel comparative field trial against conventional identification methods. For 200 strains, the standard method and PGUA/indole test were performed from the primary culture plate. The remaining 61 (23.4%) required subculture before testing. Sensitivity, specificity, positive predictive value, and negative predictive value of PGUA/indole test in the screening for E. coli were, respectively, 94.1%, 100%, 100%, and 87.1%. In our experience, PGUA/indole test is a rapid, precise, simple-to-perform, and economical method for screening E. coli. However, the need for a large inoculum may limit its application on primary cultures.

Bacteriological Techniques↗
PubMedDate not supplied

[Can we increase phosphate removal with conventional hemodialysis?].

BACKGROUND: The aim of the present study was to investigate the effect of different dialysate buffer and glucose concentrations, membrane surface (S) bigger than 2 m2 and increased dialysate flow (Qd) in phosphate (P) removal. METHODS: A. First phase (5 patients): the following variations in dialysate composition were introduced. A: glucose 1.60 g/L, bicarbonate: 39 mEq/L, acetate 4 mEq/L, B: glucose 1.5 g/L bicarbonate 17 mEq/L, acetate 10 mEq/L; C: glucose 0, bicarbonate: 39 mEq/L, acetate 4 mEq/L; and D: glucose 0, bicarbonate 17 mEq/L, acetate 10 mEq/L. B. Second phase (14 patients): variations in S and Qd were: 1. Qd: 500 mL/min + Hemophan 2 m2, 2. Qd: 500 mL/min + Hemophan 2.6 m2, 3. Qd: 750 mL/min + Hemophan 2 m2, 4. Qd: 750 mL/min + Hemophan 2.6 m2. RESULTS: Comparing HDs performed with low bicarbonate (B and D) respect to current buffer formulations (A and C), total P removal was 997.3 (+/- 237.3) vs 882 (+/- 216.1) mg (p NS). No differences were found by grouping the sessions according to glucose concentration. There were no significant differences in total phosphate removal between the two different S or Qd. The most important predictive factor of total P removal was the initial P and 2 hours serum P concentration, and PTH concentration. CONCLUSIONS: i) Removal of P is better predicted by pre-dialysis P serum concentration; ii) P removal was not affected by the changes in bicarbonate and glucose concentration in the dialysate; iii) the increase of the dialyzer area between 2 and 2.6 m2 augments Kt/V, but without influencing P elimination; iv) a higher Qd does not determine significant differences in P removal and v) higher PTH is associated with a higher P elimination.

Adult↗