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Biomedical subjects

D Hernandez

Publications and source records attributed to D Hernandez.

At least 73 records · Page 4Linked to original sources

Fine mapping of the chromosome 11q22-23 region using PFGE, linkage and haplotype analysis; localization of the gene for ataxia telangiectasia to a 5cM region flanked by NCAM/DRD2 and STMY/CJ52.75, phi 2.22.

Using pulsed-field gel electrophoresis, and a range of different enzyme digests, we have established that both markers of each of the pairs CJ52.208/YNB3.12, NCAM/DRD2, and STMY/CJ52.75, on chromosome 11q22-23, show physical linkage on a single DNA fragment. We have also shown, using genetic linkage and haplotype analyses, that these markers lie within a region of approximately 18cM, which, it has been shown previously, is likely to contain the A-T gene. The relative positions of these marker loci, and the distance between them was determined in order to construct a detailed map which has allowed a more precise localization of the A-T gene. We have shown that in pairwise linkage analysis the strongest support for linkage to the A-T gene was with the STMY/CJ52.75 locus (Z = 5.59, theta = 0.0). A three-point analysis using the results from STMY/CJ52.75 and the closely linked marker phi 2.22 gave Z = 5.55, theta = 0.03. Despite persisting evidence of some linkage to Thy-1 our results are consistent with the existence of a single A-T locus on chromosome 11q22-23 and our best estimate of the position of this locus places it between NCAM/DRD2 and (STMY/CJ52.75, F2.22) (Z = 6.74), a region of approximately 5cM in males.

Ataxia Telangiectasia↗

Monoclonal analysis of fine-needle aspiration biopsy in kidney allografts.

To evaluate changes in T-lymphocyte subsets and DR expression on tubular cells, 74 fine-needle allograft aspirates (FNAB) were evaluated in 31 patients with cadaver kidney transplants. Monoclonal antibodies against T helper CD4+, cytotoxic/suppressor CD8+, and HLA-DR were used with an indirect alkaline-phosphatase-staining technique. Cases with acute rejection (n = 11) showed a significant increase of CD8+: CD4+ ratio versus those with stable function (n = 21), acute tubular necrosis (n = 10) or CsA toxicity (n = 7) (ANOVA F = 10; P less than 0.01). Cases with chronic rejection or CMV infection showed no differences in the CD8+: CD4+ ratio with the other groups. DR expression on tubular cells was frequently found in cases of acute rejection, chronic rejection and CMV (73%, 66%, and 43% respectively), occasionally found in CsA toxicity (14%), but never seen in controls or ATN. Both tests, the CD8+: CD4+ ratio and the DR expression on tubular cells, had a high sensitivity and specificity in differentiating acute rejection versus controls, acute tubular necrosis, and CsA toxicity. When both tests are taken together no case without rejection showed a CD8+:CD4+ ratio greater than 1.6 and DR expression on tubular cells. Cases with acute rejection who lost the graft (n = 6) had a CD8+:CD4+ ratio significantly greater than those who responded to antirejection therapy (n = 5) (t = 2.9; P less than 0.05).

Antibodies, Monoclonal↗

Occupational asthma caused by inhalation of casein.

Solutions of casein are usually sprayed over the leather in the final stage of tanning. We present the result of a thorough investigation of an atopic tannery worker with occupational asthma. Data from clinical record and metacholine challenges support the occupational source of the patient's symptoms, and positive bronchial challenge with casein clearly defines it as the specific aetiological agent. The presence of specific IgE suggests a hypersensitivity type I mechanism. In the tanning process chromium salts, paraphenylendiamine and formaldehyde have all been included as specific aetiological agents of asthma, but occupational asthma induced by inhalation of casein has not previously been reported.

Adult↗

Oxygen inhibition of nitrate uptake is a general regulatory mechanism in nitrate respiration.

It has recently been demonstrated that oxygen inhibits nitrate uptake by denitrifying Pseudomonas aeruginosa. The purpose of the present investigation was to determine if this novel mechanism of regulation is universal for the regulation of nitrate respiration in other widely divergent species of bacteria. Nitrate transport by whole cell suspensions was completely and reversibly inhibited in 11 out of 12 species tested, whereas nitrate reduction by cell-free extracts was not affected by oxygen or was only partially inhibited in some cases. These results indicate that oxygen inhibition of nitrate uptake is a general regulatory phenomenon.

Enterobacter↗

Pelvic exenteration for carcinoma of the cervix: analysis of 252 cases.

We present the results of 252 pelvic exenterations for primary and recurrent carcinoma of the cervix at the Hospital General de Mexico, a tertiary-care institution for the indigent. Emphasis is placed on the morbidity and mortality of the procedure in relation to patient selection. In underdeveloped countries, where early detection of cervical cancer is a rare event, pelvic exenteration must continue in the armamentarium of physicians; it can be associated with gains in the quality of life, with long-term survival, with effective rehabilitation, and possibly with cures.

Adult↗

Olive (Olea europea) pollen allergens--II. Isolation and characterization of two major antigens.

A dialyzed extract of olive (Olea europea) pollen was fractionated by anion exchange chromatography on DEAE-Sepharose CL-6B using a discontinuous gradient of ammonium bicarbonate. The most important protein allergen was obtained from the 0.3 M fraction after gel filtration on Sephadex G-100 and separation by lentil-lectin Sepharose-4B. The major allergen of olive pollen was contained in the effluent and was designated Olea Antigen I. This material inhibited the RAST activity of 15 patients' sera that were tested. Analytical IEF demonstrated a major band at pH 5.3 and two minor ones at pH 5.6 and 5.0. When these were run into SDS-polyacrylamide gel electrophoresis in a second dimension, all were separated into two bands of mol. wt 17 and 19 K. A second protein, which is the next most important allergen, Olea Antigen II, was obtained from the 0.5 M fraction by chromatofocusing in a 4-7 pH range followed by filtration on Bio-gel P-30. Olea Antigen II had a mol. wt of 8 K as assessed by SDS-PAGE. IEF analysis displayed one main band at pH 3.6 and two minor bands at pH 3.8 and 4.0, respectively. OL-1, an anti-Olea europea monoclonal antibody (MAb) previously reported by us Lauzurica et al. (1988) reacted with the 17 and 19 K antigens from the crude extract and with Olea Antigen I but not with Olea Antigen II.

Allergens↗

Dynamic continuity of cytoplasmic and membrane compartments between plant cells.

Fluorescence photobleaching was employed to examine the intercellular movement of fluorescein and carboxyfluorescein between contiguous soybean root cells (SB-1 cell line) growing in tissue culture. Results of these experiments demonstrated movement of these fluorescent probes between cytoplasmic (symplastic) compartments. This symplastic transport was inhibited with Ca2+ in the presence of ionophore A23187, and also with the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Both of these agents have previously been demonstrated to inhibit gap junction-mediated cell-cell communication in animal cells. In a companion experiment, a fluorescent phospholipid analogue, N-4-nitrobenzo-2-oxa-1,3-diazole phosphatidylcholine (NBD-PC), was incorporated into soybean cell membranes to examine whether dynamic membrane continuity existed between contacting cells, a transport route not existing between animal cells. Photobleaching single soybean cells growing in a filamentous strand demonstrated that phospholipid did exchange between contiguous cells.

Calcium↗

Effect of a rotating bed on the incidence of pulmonary complications in critically ill patients.

The risk of nosocomial pneumonia and atelectasis is high among critically ill immobilized patients. We hypothesized that continuous turning on the kinetic treatment table would reduce their incidence. Sixty-five critically ill patients, immobilized because of head injury or traction, were prospectively randomized for treatment in a conventional bed (n = 38) or the kinetic treatment table (n = 27). Patients were well matched for baseline demographic and pulmonary risk factors. Patients in the conventional bed group had a higher incidence of cigarette smoking. The combined incidence of significant atelectasis or pneumonia was higher (66%) in the conventional vs. kinetic treatment table (33%) groups (p less than .01). Atelectasis, pneumonia, adult respiratory distress syndrome, requirements for ventilator treatment, for PEEP, and for an FIO2 greater than 0.50 were not significantly different, but tended to be higher in the control group. Survival and the incidence of decubitus ulcers were similar.

Adult↗

Combined intrathecal morphine and bupivacaine for cesarean section.

The effects of adding 0.2 mg preservative-free morphine sulfate in 0.2 ml solution to hyperbaric spinal bupivacaine were evaluated in a double-blind randomized prospective study of 34 patients undergoing elective repeat cesarean section. In the control patients (n = 17), 0.2 ml saline instead of morphine was added to bupivacaine. The intrathecal morphine significantly improved intra- and postoperative analgesia, e.g., 82% of patients given morphine compared with 41% of the control patients did not require analgesic supplementation to the spinal anesthesia during surgery; postoperatively, the former patients did not request additional analgesia for 27 +/- 0.7 hours (mean +/- SEM) compared with 2 +/- 0.3 hours in the control patients. Neonatal condition was not adversely affected by this small dose of morphine administered 11 +/- 1 minutes before delivery. Combining 0.2 mg morphine with hyperbaric spinal bupivacaine for cesarean section is a safe and effective method of improving intraoperative pain relief and providing adequate prolonged postoperative analgesia.

Analgesia↗

T3-T cell receptor (Ti) complex-independent activation of T cells by wheat germ agglutinin.

The T3-Ti complex appears to play a central role in the activation of T cells by antigens and mitogens. Wheat germ agglutinin (WGA) is a unique lectin which inhibits T cell proliferation induced by mitogens, but it also induces marked IL 2 production by peripheral blood T cells. The pattern of responses induced by WGA suggests that this lectin may use a different mechanism of T cell activation other than the mechanism employed by the common T cell stimulants. We first investigated the production of IL 2 by Jurkat cells (E6-1) stimulated with WGA, before and after modulation of the surface T3-Ti complex. IL 2 production was markedly reduced after modulation of the T3 antigen from the cell surface when these cells were stimulated with PHA. In contrast, little change was observed in WGA-induced IL 2 production after modulation. Furthermore, we examined the effect of WGA on a T3-mutant of E6-1 cells (T3.1) which does not produce IL 2 in response to PHA or PHA plus PMA. WGA-stimulated T3.1 cells produced a significant amount of IL 2 with or without added PMA. In addition, a small but consistent rise in intracytoplasmic free calcium was observed when these cells were stimulated with WGA. These results demonstrate the presence of an alternative mechanism of T cell activation independent of the T3-Ti complex.

Acetylglucosamine↗

Cell growth cycle block of T cell hybridomas upon activation with antigen.

Stimulation of antigen-specific T cell hybridomas with the appropriate antigen/MHC combination, at concentrations that resulted in the secretion of the lymphokine interleukin 2, resulted in a dose-dependent decrease in both [3H]thymidine incorporation and cell growth. Flow cytometric studies demonstrated that stimulation with antigen resulted in a cell cycle block that was most evident at the G1/S border, and mixing studies revealed that bystander T cells of different antigen specificities were not affected. For at least the large majority of T cells, the G1/S cell cycle block appeared to be irreversible after 24 h of exposure to antigen. This cell cycle block may be useful as a rapid and quantitative measure of T cell hybridoma activation, as a means of selecting T cell hybridomas that have functional alterations in the reception of stimulatory signals, and may serve as a model of the induction of tolerance in immature T cells.

Animals↗

Oxygen regulation of nitrate uptake in denitrifying Pseudomonas aeruginosa.

Oxygen had an immediate and reversible inhibitory effect on nitrate respiration by denitrifying cultures of Pseudomonas aeruginosa. Inhibition of nitrate utilization by oxygen appeared to be at the level of nitrate uptake, since nitrate reduction to nitrite in cell extracts was not affected by oxygen. The degree of oxygen inhibition was dependent on the concentration of oxygen, and increasing nitrate concentrations could not overcome the inhibition. The inhibitory effect of oxygen was maximal at approximately 0.2% oxygen saturation. The inhibition appeared to be specific for nitrate uptake. Nitrite uptake was not affected by these low levels of aeration, and nitrite reduction was only partially inhibited in the presence of oxygen. The regulation of nitrate respiration at the level of transport by oxygen may represent a major mechanism by which the entire denitrification pathway is regulated in P. aeruginosa.

Dose-Response Relationship, Drug↗

Electrophysiological effects of bupivacaine on transitional pacemaker cells of the frog heart.

The effects of bupivacaine, a long-acting local anesthetic, were studied on action potentials of spontaneously beating sinus venosus of the frog Caudiverbera caudiverbera. The study was accomplished by recording transmembrane potentials of pacemaker transitional cells. Action potentials of these cells are known to have both a fast and a slow ionic component. Bupivacaine (1 X 10(-5) M to 1 X 10(-4) M) caused a reversible negative chronotropic effect accompanied by decrease of diastolic depolarization rate and prolongation of sinus cycle length. This drug also markedly depressed rate of rise [(dV/dt)max] of action potentials. Tetrodotoxin (TTX) (1 X 10(-7) M) induced similar effects than bupivacaine. In atropine-treated sinus venosus the effect of bupivacaine on spontaneous rate of firing was less intense. However, in cells exposed to both drugs, a greater depression of (dV/dt)max and of other electrophysiological parameters such as action potential amplitude, overshoot and maximum diastolic potentials was observed. Depression of phase 4 depolarization seems difficult to be explained in view of the complex nature of the ionic currents underlying pacemaker depolarization. Reduction of (dV/dt)max caused by bupivacaine and mimicked by TTX was ascribed to blockade of sodium channels. It was concluded that bupivacaine by decreasing (dV/dt)max of transitional cells and by depressing diastolic depolarization slows down conduction of impulses arising from primary cells and that this action may contribute to its cardiac toxicity.

Action Potentials↗

Association of sinus histiocytosis with massive lymphadenopathy and idiopathic hypereosinophilic syndrome.

A patient was evaluated because of edema, pruritus and generalized painless lymphadenopathy. Laboratory tests showed marked eosinophilia without known etiology. CT scan of abdomen revealed multiple lymph nodes in retroperitoneal area. Lymph node biopsy was reported as sinus histiocytosis, bone marrow biopsy showed hypercellularity with marked infiltration of normal eosinophils. During his admission he developed Coombs positive hemolytic anaemia. Once he was stable, a laparotomy was performed and the patient died two days later because of septic shock. Autopsy revealed sinus histiocytosis with massive lymphadenopathy (SHML) with extranodal involvement of duodenum, spleen and prostate; septic liver and spleen, pyelonephritis, marked infiltration of eosinophils in lymph nodes, spleen, liver duodenum and lungs. To the best of our knowledge, this is the first case report of the association of SHML and Idiopathic Hypereosinophilic Syndrome (HES).

Autopsy↗

Gas chromatographic assay for in vitro complementation of Pseudomonas aeruginosa mutants deficient in nitrate reduction.

An electron capture gas-chromatographic technique was developed to continuously measure nitrate (NO3-) reduction during in vitro complementation tests with extracts from Pseudomonas aeruginosa mutants deficient in both assimilatory and dissimilatory nitrate reduction as a result of a single genetic mutation. The procedure involves coupling nitrate reduction to nitrous oxide (N2O) evolution via a series of reactions specific to the denitrification pathway. The assay was dependent on nitrate concentration, and optimal activity was obtained with a final concentration of 0.2% potassium nitrate. The reduction exhibited a stoichiometry of 2:1 (NO3-/N2O), and succinate was the best electron source for the reaction, followed by glucose, pyruvate, and malate. The technique can also be used for continuously monitoring nitrate reduction. The optimal nitrite concentration in the nitrite reductase assay was 0.025%. The initial complementation studies of mutant extracts demonstrated that at least two genes are shared between the two nitrate reduction pathways in P. aeruginosa.

Chromatography, Gas↗