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Biomedical subjects

F Ojeda

Publications and source records attributed to F Ojeda.

At least 19 recordsLinked to original sources
PubMed2004

Ultraviolet exposure of thymocytes: selective inhibition of apoptosis.

PURPOSE: To evaluate selective effects of ultraviolet (UV) irradiation on spontaneous and induced apoptosis in freshly extracted mice thymocytes. MATERIALS AND METHODS: Cells were exposed to UV radiation with emission peaks of 365 nm (UVA) exposures of 1620-10200 J m(-2), of 312 nm (UVB) exposures of 34-1620 J m(-2) or of 254 nm (UVC) exposures of 1.5-1620 J m(-2), and incubated for 5.5 h with or without hydrocortisone, phorbol-12-myristate-13-acetate or anti-Fas antibody. Additionally, cells were irradiated with gamma-rays (5 Gy) before UVB exposure (408 J m(-2)) at different times. Apoptosis was quantified by DNA fragmentation. RESULTS: Up to an irradiation of 5000 J m(-2), UVA exposure did not show any effect on thymocyte apoptosis, while at 10200 J m(-2) irradiation, considerable DNA fragmentation was observed. In contrast, UVB and UVC irradiation clearly inhibited natural and cortisone-induced apoptosis. Moreover, UVB inhibited apoptosis triggered by phorbol-12-myristate-13-acetate and gamma-irradiation, but not by anti-Fas antibody. CONCLUSIONS: The response of mouse thymocytes in culture to UV irradiation strongly depends on the wavelength used. It is suggested that either a survival or an apoptotic pathway occurs depending on the physiological state of the cell, spectral composition of the UV light and cell type. The possible involvement of extracellular signal-regulated kinase and stress-activated protein kinase/c-Jun N-terminal kinase in the apoptotic pathway is discussed.

Animals↗
PubMed2000

Dynamics of rough interfaces in chemical vapor deposition: experiments and a model for silica films

We study the surface dynamics of silica films grown by low pressure chemical vapor deposition. Atomic force microscopy measurements show that the surface reaches a scale invariant stationary state compatible with the Kardar-Parisi-Zhang (KPZ) equation in three dimensions. At intermediate times the surface undergoes an unstable transient due to shadowing effects. By varying growth conditions and using spectroscopic techniques, we determine the physical origin of KPZ scaling to be a low value of the surface sticking probability, related to the surface concentration of reactive groups. We propose a stochastic equation that describes the qualitative behavior of our experimental system.

Journal Article↗
PubMed1998

Methyl-beta-cyclodextrins and liposomes as water-soluble carriers for cholesterol incorporation into membranes and its evaluation by a microenzymatic fluorescence assay and membrane fluidity-sensitive dyes.

A variety of methods to incorporate cholesterol into lipid membrane systems have been applied with varying success. We tested an incorporation method based on cholesterol-loaded methyl-beta-cyclodextrins and compared it to a method that uses cholesterol-loaded liposomes. With methyl-beta-cyclodextrin, we increased the cholesterol content in microsomal membranes to almost the fourfold of the original content. With cholesterol-loaded liposomes instead, we achieved an elevation of 140%. Short incubation times and well-defined carrier properties favor the beta-cyclodextrin method. For direct detection of membrane cholesterol, we slightly modified a microenzymatic fluorescence assay originally developed for precise cholesterol detection in serum. Without the need to perform lipid extraction, this assay was reliable for cholesterol detection in liposomes and in microsomes. Additionally, we compared the sensitivity of the fluidity-sensitive fluorescent dyes pyrene, pyrene-methanol, bis-pyrene, 1-6-phenyl-1,3,5,-hexatrien, and 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5,-hexatrien in order to detect cholesterol indirectly by the dynamically relevant changes exerted on lipid matrices. These dyes differ not only in their membrane location but also in their dynamical behavior. We calibrated the dyes in liposomes of defined cholesterol content and used the most suited ones to follow and quantify the cholesterol incorporation into liposomal and microsomal membranes.

Animals↗
PubMed1998

Cholesterol induced variations of membrane dynamics related to the induction of apoptosis in mouse thymocytes.

PURPOSE: To evaluate the involvement of cholesterol induced variations of membrane dynamics in mouse thymocyte apoptosis. MATERIALS AND METHODS: Membranes of thymocytes of RK mice were enriched with cholesterol using methyl-beta-cyclodextrins as carriers. Spontaneous apoptosis was compared with apoptosis induced either by X-irradiation, by treatment with dexamethasone (DEX), and by phorbol-12-myristate-13-acetate (PMA). Apoptotic cells were quantified by means of flow cytofluorometry. RESULTS: Small amounts of incorporated cholesterol enhance the cellular sensitivity for spontaneous apoptosis whereas larger amounts of incorporated cholesterol protect against spontaneous apoptosis and apoptosis induced by irradiation, DEX, or PMA. CONCLUSIONS: Cholesterol exerts specific rigidity effects on lipid membranes which have been shown to be involved in thymocyte apoptosis. The general effect of higher concentrations of cholesterol protecting against apoptosis hints towards a central protective mechanism. This study believes that either cholesterol paralyses great parts of the cell metabolism or that the apoptotic chain reaction is interrupted at a central point due to protection of membrane lipid regions from oxidative stress.

Animals↗
PubMed1995

Induction of apoptosis in thymocytes: new evidence for an interaction of PKC and PKA pathways.

The second messenger cascades connected to PKC and PKA are involved in the induction of apoptosis. Here we study the interaction of those two second messenger pathways with respect to the induction of apoptosis by stimulation or inhibition. The stimulators used were phorbol dibutyrate for PKC and one of the cAMP agonists Sp-5,6 DCl-cBIMPS or Sp-cAMP for PKA. The inhibitors used were staurosporin for PKC and the cAMP antagonist Rp-cAMPS for PKA. We found a synergism between both second messenger systems with regard to the induction of apoptosis in thymus lymphocytes.

Animals↗
PubMed1994

Heterogeneity of microsomal membrane fluidity: evaluation using intrinsic tryptophan energy transfer to pyrene probes.

Membrane fluidity measurements based on excimer formation of pyrene and pyrene derivatives as a measure of lateral diffusion yield a decreased fluidity in the presence of proteins [1-3]. It was the aim of our study to investigate whether the reduced excimer formation is due to a rigidifying effect of proteins on the whole membrane or if the fluorophore mobility is mainly hindered in the immediate protein environment. Resonance energy transfer in microsomal membranes between intrinsic tryptophan residues and pyrene were used to study the excimer formation rate in the vicinity of proteins. The excimer-to-monomer fluorescence ratio at excitation via resonance energy transfer is lower than that observed for the direct excitation. The results suggest that, because of a reduced fluidity in the neighbourhood of proteins, pyrene and pyrene fatty acids do not diffuse homogeneously in the membrane plane. A fluidity gradient exists from the membrane proteins to the bulk lipid.

Animals↗
PubMed1994

Membrane fluidity of microsomal and thymocyte membranes after X-ray and UV irradiation.

A brief literature review shows that ionizing radiation in biological membranes and in pure lipid membranes causes malondialdehyde formation, indicating lipid peroxidation processes. With respect to membrane fluidization by ionizing radiation, in pure lipid membranes rigidization effects are always reported, whereas contradictory results exist for biological membranes. Starting from the assumption that membrane proteins at least partly compensate for radiation effects leading to a rigidization of membrane lipid regions, pig liver microsomes, as a representative protein-rich intracellular membrane system, were irradiated with X-rays or UV-C with doses up to 120 Gy at a dose rate of 0.67 Gy min-1 and up to 0.73 J cm-2 at an exposure rate of 16.2 mJ cm-2 min-1, respectively. For both irradiation types a weak but significant positive correlation between malondialdehyde formation and membrane fluidity is revealed throughout the applied dose ranges. We conclude that the membraneous protein lipid interface increases its fluidity under radiation conditions. Also, thymocyte ghosts showed an increased fluidity after X-ray irradiation. Fluidity measurements were performed by the pyrene excimer method.

Animals↗
PubMed1994

Radiation induced membrane changes and programmed cell death: possible interrelationships.

A short review of the evidence that lymphocyte membranes are a target for the initiation of irradiation induced programmed cell death (PCD) is given. It is assumed that for lymphocytes PCD represents an essential physiological mechanism in order to prevent degeneration of the biological system involved. Initiation of PCD can be obtained by a pharmacological activation as well as with irradiation. In both cases, protein kinase-C (PKC) is involved in the signal transduction from the cellular membrane to the nucleus where, by means of a metabolically active process, DNA fragmentation is induced. It is hypothesized that processes connected to lipid peroxidation in the cell membrane constitute a primary effect of irradiation induced PCD, where membrane fluidization or a compensatory process aimed to the maintenance of membrane fluidity (membrane homeoviscosity hypothesis) are likely to be involved.

Animals↗
PubMed1992

A flow-cytometric method to study DNA fragmentation in lymphocytes.

A method to measure DNA fragmentation cell by cell in a cell population was implemented based on acridine orange procedure to determine DNA content of single cells by flow cytometry. Using this method it can be observed that the fragmentation process induced by irradiation in thymic cells occurs in a fraction of the population, thus indicating that this process is not evenly distributed over the total population, and that it corresponds to a fast phenomenon in which the cells suddenly lose DNA material.

Acridine Orange↗
PubMed1992

Role of protein kinase-C in thymocyte apoptosis induced by irradiation.

The role of protein kinase C in radiation-induced death of thymocytes was studied. For this purpose murine thymocytes were irradiated and incubated for 6 h at 37 degrees C and afterwards the fraction of fragmented DNA was measured. Results indicate that radiation-induced DNA fragmentation can be prevented by adding the protein kinase C inhibitor H-7 or staurosporine to the thymocytes during incubation time. Incubation of irradiated cells with HA-1004, an inhibitor of cAMP-dependent protein kinase, with a minor effect on protein kinase C did not affect the DNA fragmentation induced by irradiation. Incubation of cells with phorboldibutyrate gave a dose-dependent induction of DNA fragmentation. This effect can be inhibited by staurosporine. These results suggest that radiation-induced DNA fragmentation is an active cellular process in which protein kinase C plays an important role.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1991

Rise in thymocyte number and thymulin serum level induced by noise.

A high level of noise is known to induce important changes in the immune system. In this work, the effect of sound stress on the circulating level of thymulin and on the cellularity of the thymus gland was studied. The experiments were done in RK mice exposed to a noise level of 100 dB for a period of 1 h. Following the noise exposure, the animals were bled at different times for thymulin titration, or killed in order to evaluate the number of cells and the weight of each thymus. The results indicate that young mice exposed to the stressor stimulus show an increase in serum thymulin titre, and at the same time they show an increment in thymus weight and in thymocyte number compared to control. These results support a new argument in favour of the theory of a central nervous system control on the thymus function.

Age Factors↗
PubMed1991

Radiation-induced surface IgG modulation: protein kinase C involvement.

Although radiation-induced loss of surface IgG (s-IgG) expression on murine B cells is known to be dependent on intact energy metabolism and integrity of the cytoskeleton, the exact mechanism of this radiation effect is not known. Evidence reported here shows that inhibition of protein kinase C (PKC) by H-7 impairs the radiation-induced s-IgG modulation, whereas addition of HA-1004, which preferently inhibits c-AMP-dependent protein kinase, shows only minor effects. On the other hand PMA, a PKC activator, mimics the radiation effect, and H-7 but not HA-1004 inhibits the PMA-induced loss of s-IgG expression. Therefore it is suggested that PKC is involved in the modulation of s-IgG induced by irradiation on B cells. The possibility of membrane participation in this event is discussed.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1991

Hospital-acquired diarrhea in adults: a prospective case-controlled study in Mexico.

OBJECTIVE: To know the incidence, etiology, risk factors, morbidity, and mortality of nosocomial diarrhea in adults. DESIGN: Nested case-control study, matched by service, length of stay, date of admission, and presence of leukopenia and/or the acquired immunodeficiency syndrome (AIDS). Cases were those who developed nosocomial diarrhea. Controls were those who did not develop nosocomial diarrhea during a comparative period nor during the next ten days. Stool samples were processed in search for parasites, yeasts, bacteria, and rotavirus. SETTING: Third-level referral center, in Mexico City, Mexico, for general internal medicine and surgical problems. PATIENTS: Eligible subjects were all new admissions to the hospital from November 1987 to September 1988. Reasons for exclusion were presence of chronic diarrheal disease or melena. There were 115 cases and 111 controls. RESULTS: Overall risk of acquiring nosocomial diarrhea was 5.5%, or 1.8 episodes per 100 patient-weeks. A potential pathogen was found in 59%. Yeasts and Entamoeba histolytica were the most frequently isolated pathogens. Mortality in cases was 18%, as compared with 5% in controls (p less than .01). Multivariate analysis showed enteral feeding, recent enemas, presence of Candida species, use of antacids/H2-blockers, and presence of nasogastric tubes as significant risk factors for nosocomial diarrhea. CONCLUSIONS: Diarrhea is a common complication in hospitalized patients. It occurs more often than previously suspected and is linked with a substantial mortality. The spectrum of etiologic agents is different from that reported in pediatric hospitals. Given that nosocomial diarrhea may constitute, at least, a marker of severity of illness, it should receive more attention in general hospitals.

Adult↗
PubMed1990

Protein kinase-C involvement in thymocyte apoptosis induced by hydrocortisone.

The involvement of protein kinase-C in thymocytes death induced by hydrocortisone was studied. Thymus cells were incubated 6 hr or in the presence of hydrocortisone, labeled with Acridine orange, and the DNA content of each nuclei was estimated by cytofluorimetry. The results indicate that hydrocortisone-induced DNA fragmentation can be prevented by adding the protein kinase-C inhibitor H-7 to the cell suspension. Incubation of the H-A 1004, an inhibitor of c-AMP-dependent protein kinase, with low effect on on protein kinase-C, did not interfere with the cortisone-mediated DNA fragmentation. Therefore, it can be concluded that protein kinase-C plays an important role in the process of lympholysis mediated by corticoids.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1990

Recovery of HIV antigen in peritoneal dialysis fluid.

We investigated the presence of HIV antigen in dialysis fluid of patients with end-stage renal disease (ESRD) undergoing continuous ambulatory peritoneal dialysis (CAPD), previously known to be infected with this virus. Sixteen adult patients and 6 adult volunteers were included in the study in 4 groups as follows: Group A: 3 patients on CAPD, previously known to be positive for serum HIV antibodies; Group B: 7 patients on CAPD, serum HIV negative; Group C: 6 AIDS patients without renal disease; and Group D: 6 healthy volunteers. Of the 3 patients of Group A, the HIV-1 Ag was positive in dialysis fluid in only 2. In 1, serum Ab and Ag were present, while in the others only serum Ab was detected. The samples from Group B were all negative for the viral antigen in dialysis fluid. We conclude that dialysis fluid of HIV-infected patients may contain the Ag and is therefore potentially infective. The presence of the HIV antigen was not constant, and was not related to antigenemia. It is possible that the presence of the Ag depends on local factors that influence viral replication or to alterations in the permeability of the peritoneal membrane. We discuss other possible factors that could influence the presence of viral Ag in peritoneal dialysis fluid.

Acquired Immunodeficiency Syndrome↗
PubMed1986

Control of emergence of multi-resistant gram-negative bacilli by exclusive use of amikacin.

Results of a three-year prospective study of amikacin as the only aminoglycoside used at the Instituto Nacional de la Nutrición "Salvador Zubirán" are presented. During the initial three-month baseline period, resistance to amikacin, gentamicin, and tobramycin among 870 gram-negative bacterial isolates was 3.2 percent, 17.4 percent, and 11.2 percent, respectively. In this period, the overall consumption of aminoglycosides was 69 percent for gentamicin, 30.5 percent for amikacin, and 0.5 percent for tobramycin. In the following period of exclusive amikacin use, sensitivity patterns of 9,344 gram-negative strains isolated over three years were recorded. During this period, amikacin constituted 99.3 percent of all aminoglycosides used. The percentage of gentamicin-resistant gram-negative strains declined to 7.4 percent (p less than 0.0001), whereas the percentage of amikacin-resistant strains did not change significantly. Quarterly trend analysis of aminoglycoside-resistant strains also demonstrated a significant decrease in gentamicin resistance (p less than 0.005) and an overall steady state of amikacin resistance. It is concluded that the exclusive use of amikacin was not accompanied by a significant increase in amikacin resistance during a three-year period, and may even lead to a decrease in resistance to gentamicin and tobramycin among most gram-negative organisms.

Amikacin↗