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Biomedical subjects

C Gutierrez

Publications and source records attributed to C Gutierrez.

At least 163 records · Page 9Linked to original sources
PubMed1989

Sequence of an osmotically inducible lipoprotein gene.

The osmB gene of Escherichia coli, whose expression is induced by elevated osmolarity, was cloned and physically mapped to a 0.65-kilobase-pair NsiI-HincII DNA fragment at 28 min on E. coli chromosome. The OsmB protein was identified in minicells expressing the cloned gene. The nucleotide sequence of a 652-base-pair chromosomal DNA fragment containing the osmB gene was determined. The open reading frame encodes a 72-residue polypeptide with an Mr of 6,949. This reading frame was confirmed by sequencing the fusion joint of an osmB::TnphoA gene fusion. The amino-terminal amino acid sequence of the open reading frame is consistent with reported signal sequences of exported proteins. The sequence around the putative signal sequence cleavage site, Leu-Ser-Ala-Cys-Ser-Asn, is highly homologous to the consensus sequence surrounding the processing site of bacterial lipoproteins. The presence of a lipid moiety on the protein was confirmed by demonstrating the incorporation of radioactive palmitic acid and inhibition of processing by globomycin. Preliminary localization of the authentic OsmB protein was determined in minicells harboring a plasmid that carries the NsiI-HincII fragment; it was primarily in the outer membrane. Surprisingly, an osmB mutant carrying the osmB::TnphoA insertion mutation was more resistant to the inhibition of metabolism by high osmolarity than the parent strain was.

Amino Acid Sequence↗
PubMed1989

Origin auxiliary sequences can facilitate initiation of simian virus 40 DNA replication in vitro as they do in vivo.

Initiation of simian virus 40 (SV40) DNA replication is facilitated by two auxiliary sequences that flank the minimally required origin (ori) core sequence. In monkey cells, the replication rate of each of the four ori configurations changed with time after transfection in a characteristic pattern. This pattern was reproduced in an extract from SV40-infected monkey cells by varying the ratio of DNA substrate to cell extract; DNA replication in vitro depended on ori auxiliary sequences to the same extent as they did in vivo. Facilitation by ori auxiliary sequences was lost at high ratios of DNA to cell extract, revealing that the activity of these sequences required either multiple initiation factors or a molar excess of one initiation factor bound to ori. This parameter, together with ionic strength and the method used to measure DNA replication, determined the level of facilitation by ori auxiliary sequences in vitro. The activity of ori auxiliary sequences was not diminished in vivo or in vitro by increasing amounts of large tumor antigen. Therefore, ori auxiliary sequences promoted initiation of replication at some step after tumor antigen binding to ori. Furthermore, although cellular factors could modulate the activity of ori auxiliary sequences in vitro, these factors did not appear to involve nucleosome assembly because no correlation was observed between the number of nucleosomes assembled per DNA molecule and facilitation by ori auxiliary sequences. These results demonstrate that SV40 ori auxiliary sequences can function in vitro as they do in vivo and begin to elucidate their role in initiating DNA replication.

Animals↗
PubMed1989

Repolarization of Na+-K+ pumps during establishment of epithelial monolayers.

Madin-Darby canine kidney (MDCK) cells plated at confluence and incubated for 20 h in low (5 microM) Ca2+ have no tight junctions (TJs), and their Na+-K+-ATPase is randomly distributed over the surface. On transfer to normal Ca2+ levels (1.8 mM) ("Ca2+ switch"), TJs and transepithelial resistance develop quickly, trapping a considerable fraction (35%) of the surface Na+-K+-ATPase on the apical (incorrect) side. This misplaced enzyme is subsequently removed from this region or inactivated, demonstrating that polarization proceeds despite TJs. Simultaneously, the amount of Na+-K+-ATPase on the basolateral side increases in a higher proportion (125%), than could be accounted for by relocation of the misplaced apical enzyme. This incorporation is prevented by cycloheximide, ammonium chloride, primaquine, or chloroquine, suggesting that Na+-K+-ATPase originates in an intracellular pool and that its surface insertion requires synthesis of new enzyme or of a protein factor, since it is carried to the surface membrane through a mechanism of exocytosis. In summary, asymmetric distribution of ion pumps depends 1) on polarized insertion of Na+-K+-ATPase as well as 2) on removal or inactivation of misplaced enzyme.

Animals↗
PubMed1989

Primate cardiac allo-and xenotransplantation: modulation of the immune response with photochemotherapy.

Survival of heterotopic allo- and xenografts was studied in a primate cardiac transplantation model. Initially, animals were presensitized to donor xenoantigens by blood transfusion and treated with strategies designed to elicit pretransplant immunosuppression. These animals underwent rapid rejection whether treated with ciclosporin and conventional immunosuppression, autologous sera, sensitized lymphocytes, or photochemotherapy. In a nonsensitized xenograft study, xenografts were maintained for prolonged periods with a combination of ciclosporin and photochemotherapy. All animals treated with photochemotherapy demonstrated periodic suppression of the mixed leukocyte culture response to their donor. Inhibition of lymphocytotoxic antibody was achieved only in animals treated with both ciclosporin and photochemotherapy. Xenosensitized animals were secondarily exposed to donor alloantigens. These animals were successfully allografted. Ciclosporin therapy was terminated and the animals maintained by photochemotherapy alone. This preliminary trial indicates that a combination of ciclosporin and photochemotherapy may be of value in maintaining concordant xenografts and preventing antibody mediated rejection.

Animals↗
PubMed1989

[Tetralogy of Fallot associated with total anomalous drainage of the pulmonary veins. Report of 2 cases].

The authors report on two cases of tetralogy of Fallot (TF) associated with total anomalous pulmonary venous return to the coronary sinus. The clinical manifestations in both children were the same as of classical TF and diagnosis was confirmed by cineangiography. The patients underwent total surgical correction and the first one died in the immediate post-operative period. The second patient had an uneventful recovery. The rarity of this association is the main reason of this report where clinical and surgical findings are discussed.

Angiocardiography↗
PubMed1988

Effect of carbenoxolone on glucose metabolism in rat adipose tissue.

Carbenoxolone significantly decreased the glucose uptake and the incorporation of glucose into triglycerides and CO2 in rat epididymal fat pads. The effect produced by insulin on these metabolic pathways was reduced when adipose tissue was incubated with insulin in the presence of carbenoxolone (10(-3) M). On the other hand the drug (10(-3) M) produced a decrease in cyclic AMP concentration in adipose tissue similar to that produced by insulin (100 ng/ml).

Adipose Tissue↗
PubMed1988

The B-cell activation pathway in human systemic lupus erythematosus: imbalanced in vitro production of lymphokines and association with serum analytical findings.

Recent knowledge of B-lymphocyte physiology has clarified the role of T cell-derived lymphokines in clonal proliferation and differentiation of B-cell responses. Lymphokine production was analyzed in 19 systemic lupus erythematosus (SLE) patients and sex- and age-matched controls in relation to clinical activity and steroid treatment. When in vitro production of interleukin-2 (IL-2) and B-cell growth factor (BCGF) was tested, both activities were found to be diminished in the group of patients (P less than 0.01), while B-cell differentiation factor (BCDF) activity was higher in this group with respect to normal controls (P less than 0.01). Interestingly enough, this in vitro BCDF synthesis was positively correlated with clinical activity regardless of low-dose steroid treatment. A correlation was also found between BCDF production and the levels of IgG (r = 0.64, P less than 0.01), anti-DNA antibodies (r = 0.52, P less than 0.05), and the IgG/IgM ratio (r = 0.7, P less than 0.01) in serum. Implications of these abnormal T-lymphocyte functions in SLE with respect to in vivo B-cell function are discussed.

Adolescent↗
PubMed1988

Presence of an autoantibody against a Golgi cisternal membrane protein in the serum and cerebrospinal fluid from a patient with idiopathic late onset cerebellar ataxia.

Tissue and cultured cells of different species and embryological origins incubated with serum (diluted up to 10,000-fold) or cerebrospinal fluid (CSF) (6-fold dilution) from a 48-year-old female patient with idiopathic late-onset cerebellar ataxia, exhibited a bright specific perinuclear staining when studied by indirect immunofluorescence microscopy. The pattern of the staining was that characteristic of the Golgi apparatus, consisting of a crescent-shaped juxtanuclear reticulum located in the vicinity of the microtubule organizing center. Changes in location and organization of the organelle stained by the patient's serum during mitosis or after incubation of the cells with Colcemid, taxol or monensin, resulted in a disruption of the reticulum that followed the expected patterns for Golgi apparatus. The staining was specifically absorbed with Golgi cisternae-enriched membrane fractions. Finally, dot-immunoblotting studies of membrane and soluble fractions of Golgi cisternae and vesicles showed that the anti-Golgi antibody (AGA) reacted with the cytoplasmic domain of an integral membrane protein contained in the Golgi cisternae. The presence of this unusual autoantibody in an idiopathic late-onset cerebellar ataxia-bearing patient can afford some insights into the pathogenesis of these neurological diseases.

Autoantibodies↗
PubMed1988

Demonstration of cytochrome reductases in rat liver peroxisomes: biochemical and immunochemical analyses.

In this study we utilized the analytical cell fractionation approach in combination with immunoblotting techniques and immunoelectron microscopy to test for the presence of NADPH cytochrome P-450 reductase and NADH cytochrome c (b5) reductase in rat liver peroxisomes. Highly purified peroxisomes from clofibrate-treated rats exhibited both NADPH cytochrome P-450 reductase activity and NADH cytochrome c reductase activity (using cytochrome c as an electron acceptor). These activities were inhibited by the respective reductase antibodies made against the endoplasmic reticulum (ER) enzymes. Immunoblot data in combination with immunoelectron microscopy indicated that the peroxisomal NADPH cytochrome P-450 reductase is localized in the matrix of the organelle and has a subunit of molecular weight similar to that of the ER enzyme, whereas the NADH cytochrome c (b5) reductase is localized in the membranes of the peroxisomes. Again, the subunit molecular weight was similar to that of the ER enzyme. The presence of these reductases in peroxisomes further supports the role of this organelle in bile acid synthesis and cholesterol metabolism.

Animals↗
PubMed1988

Progressive cellular immune impairment leading to development of AIDS: two-year prospective study of HIV infection in drug addicts.

We have studied immunological, serological and clinical abnormalities in 264 HIV-positive and HIV-negative drug abusers. Ninety percent of the 264 drug addicts (mean age 26 +/- 0.8 years) were found to have HIV antibodies and there was a significant increase (P less than 0.01) in the frequency of HIV antibody positivity with increasing duration of exposure to parenteral drug abuse. There was a very strong correlation between the progressive decline of the mean T4+ helper/inducer cells and T4+/T8+ ratio, the low response to pokeweed mitogen and the more severe clinical manifestations of HIV infection. Impairment of delayed-type hypersensitivity reaction to recall antigens was only seen in group IV as defined by the Center for Disease Control. Within group IV, anergy was found to be highly associated (83%) in patients with opportunistic infections. All other HIV-positive addicts from groups II and III, as well as HIV-negative addicts had normal in vivo responses to test antigens. We have also analysed in a prospective follow-up lasting 6-24 months, the evolution of HIV infection in a cohort of 50 HIV-antibody-positive drug addicts. Thirty-two percent showed clinical progression and most of the drug addicts that proceeded to full-blown AIDS developed anergy (82%) prior to clinical deterioration with development of opportunistic infections. We conclude, that in seropositive drug addicts a low absolute count of helper/inducer cells (mean +/- s.e. = 243 +/- 48 cells/mm3), a low response to pokeweed mitogen and anergy are predictive markers of progression to AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1987

The use of transposon TnphoA to detect genes for cell envelope proteins subject to a common regulatory stimulus. Analysis of osmotically regulated genes in Escherichia coli.

The transposon TnphoA can be used specifically to detect bacterial genes that code for cell envelope proteins. We have used TnphoA to search for genes regulated by osmolarity in Escherichia coli. Among approximately 30,000 random insertions of TnphoA into the chromosome, we have found 700 independent fusions that produce hybrid proteins with alkaline phosphatase activity. Of these, 37 were induced after growth in a medium of high osmolarity and none was repressed. Osmo-inducible fusions of phoA were found to ompC and to a gene that is probably proU. These two genes were already known to be transcriptionally induced by osmolarity. In addition, eight other genes, designated osm, were identified and mapped on the bacterial chromosome. The expression of these genes is induced by solutes that are unable to decrease the turgor pressure applied to the envelope. One of the osm genes, osmI, is also specifically induced by glycerol, which does diffuse across the cytoplasmic membrane. The expression and osmoregulation of all the osm genes were shown to occur independently of ompR and envZ, which control the expression and osmoregulation of the ompC and ompF genes in E. coli.

Alkaline Phosphatase↗
PubMed1987

Whole bowel irrigation: experience in pediatric patients.

Whole bowel irrigation (WBI) was used in 50 children aged 16 months to 14 years in preparation for surgery (n = 30) and colonoscopy (n = 20). The patients were divided into three groups according to age: group A, 12 to 36 months; group B, 3 to 6 years; and group C, 6 to 14 years. A hydroelectrolytic solution (Na 150 mEq/L, K 30 mEq/L, Cl 130 mEq/L, HCO3 50 mEq/L) was administered to all the groups at a variable rate established according to tolerance, weight, and age. Neomycin (1 g) and erythromycin (50 mg/kg) were administered in the last liter of the solution of the patients being prepared for colorectal surgery. Vital signs (heartbeat, arterial tension, and central venous pressure) and blood parameters (Na, K, HCO3, pH, and Hct) were assessed before and after irrigation and remained stable in all the groups. The rate of administration of the solution was best tolerated in each group as follows: group A, 75 mL/kg/h; group B, 60 mL/kg/h; and group C, 45 mL/kg/h. Colonic preparation was satisfactory in 49 of the 50 cases. Postoperative infectious complications were reduced to 3.3%. WBI in pediatric patients is a fast, simple, and well-tolerated method that gets a colon in excellent condition for surgery and colonoscopy exploration and considerably decreases the number of postoperative complications.

Adolescent↗
PubMed1987

c-myc protein and DNA replication: separation of c-myc antibodies from an inhibitor of DNA synthesis.

Antibodies against human c-myc protein have been reported to inhibit DNA polymerase activity and endogenous DNA synthesis in isolated nuclei, suggesting a role for c-myc in DNA replication. Using the same antibody preparations, we observed equivalent inhibition of simian virus 40 DNA replication and DNA polymerase alpha and delta activities in vitro, as well as inhibition of DNA synthesis in isolated nuclei. However, the c-myc antibodies could be completely separated from the DNA synthesis inhibition activity. c-myc antibodies prepared in other laboratories also did not interfere with initiation of simian virus 40 DNA replication, DNA synthesis at replication forks, or DNA polymerase alpha or delta activity. Therefore, the previously reported inhibition of DNA synthesis by some antibody preparations resulted from the presence of an unidentified inhibitor of DNA polymerases alpha and delta and not from the action of c-myc antibodies.

Animals↗
PubMed1987

Complement activation by human lymphocytes from different lymphoid organs: role of sialic acid and lack of relationship to electrical surface charge.

A study was made of the complement activation (CA) capacity of human lymphocytes from lymphoid organs. Whereas thymocytes did not show complement membrane fluorescence (CMF) following incubation in normal homologous serum (NHS), a mean of 18 and 34%, respectively, of tonsil and spleen cells were positive for antihuman C3 fluorescent serum. Isolated T cells did not show CA capacity, while 62% of a purified tonsil B population possessed this capacity. The CMF was abolished by adding EDTA and when the incubation was performed in C1q and factor-D-depleted serum. Addition of EGTA, supplemented with MgCl2 to NHS failed to abolish the fluorescence, indicating that the alternative pathway was involved in the phenomenon. Removal of sialic acid with neuraminidase increased the percentage of cells showing CMF. Since sialic acid significantly contributes to the extent of net negative surface charge, we have studied whether this parameter influences the CA capacity. Tonsil B cells were separated in fractions with different electrophoretic mobility by preparative cell electrophoresis, and similar percentages of CMF-positive cells were found in all fractions. Therefore, it is likely that the capacity to activate complement is unrelated to the membrane electrical charge.

B-Lymphocytes↗
PubMed1986

Differential proliferative responses of B cells from BALB/c and autoimmune NZB mice to B-cell growth factor(s).

Highly purified B cells from NZB mice have altered responses to various stimuli which require additional costimulatory signals supplied by factors present in EL-4 supernatants when compared to age-matched control nonautoimmune strains. Both crude preparations of EL-4 supernatants as well as partially purified BSF-p1 induced peak proliferation in B cells from normal strains of mice only in the presence of another stimulatory signal, anti-mu. In contrast, B cells from autoimmune-prone NZB mice proliferated in response to B-cell growth factors, with an age-dependent variation. Splenic B cells from 16- to 22-week-old NZB mice, an age where pronounced autoimmune disease is not observed, demonstrated a near maximum proliferative response with B-cell growth factors alone. While normal B cells responded maximally to BSF-p1 in the presence of anti-mu, B cells from young adult NZB mice (16-22 weeks of age) were not further stimulated to proliferate upon the addition of anti-mu. Such NZB B cells appeared to lack the requirement for a stimulation signal delivered by anti-mu in order to respond to B-cell growth factors. These results suggested that NZB cells were partially activated in vivo in the preautoimmune state so that subliminal triggers lead to full activation.

Animals↗
PubMed1985

Caffeine induces sister-chromatid exchanges during the whole S-phase of the cell cycle.

The capacity of caffeine to induce sister chromatid exchanges (SCEs) in different cell cycle stages and the proliferation kinetics were studied. Continuous treatment with this xanthine during the whole second cycle significantly increased the baseline SCE frequency. Pulse-treatment experiments showed that the induction of SCEs by caffeine, which was dose-dependent, was restricted to the S-phase of the cell cycle without effect on G1 or G2 cells. Moreover, unlike other SCE-inducing agents, such as DNA-synthesis inhibitors and DNA-damaging agents, caffeine produced similar SCE increases in cells treated at different times throughout the S-phase. In the light of Painter's model for SCE formation and the known effects of caffeine on the DNA replication pattern, the most likely mechanism of SCE induction by caffeine is an increase in the number of DNA-replication sites.

Caffeine↗
PubMed1985

Indirect effects of the 3'-5' cyclic adenosine monophosphate binding protein (CAP) on the transcription of the malPQ operon in Escherichia coli.

Uninduced malPQ transcription, as followed by measuring beta-galactosidase expression in a strain carrying a malP-lacZ hybrid gene and grown in the absence of maltose, requires the presence of CAP. However this requirement is lost when the expression of malT, positive regulator gene of the maltose regulon, is rendered independent of CAP by a mutation in the malT promoter. This result suggests that the effect of CAP on uninduced malPQ expression is mediated through a modulation of MalT protein synthesis. The effect of CAP on the induced expression of malPQ is presumably mediated, in addition, through a modulation of the synthesis of the maltose transport system and, hence, of the entry of the inducer. Therefore the effect of CAP on malPQ expression seems to be merely indirect, and this is surprising since a CAP binding site is present at the malPQ promoter.

Carrier Proteins↗