PubMed Health⌕ Search

Biomedical subjects

R Rodriguez

Publications and source records attributed to R Rodriguez.

At least 217 records · Page 12Linked to original sources

Dimerization of the chicken progesterone receptor in vitro can occur in the absence of hormone and DNA.

We have analyzed the dimerization of two forms of the chicken progesterone receptor (cPRA and cPRB) by nondenaturing gradient gel electrophoresis and chemical cross-linking with dimethylpimelimidate (DMP). We demonstrate by these two methods that the PRs assemble in vitro into dimers in the absence of DNA, and that dimerization does not require hormone. The cPRA homodimer binds quantitatively to its cognate DNA response element in our nondenaturing gradient gel assay. DMP cross-linking confirms that both forms of the receptor (cPRA and cPRB) assemble into dimers in solution. Finally, in a standard mobility shift assay, chemically cross-linked receptors bind to the progesterone DNA response element with high affinity. We conclude that the PR contains a dimerization motif, which can promote stable subunit-subunit contacts without the presence of hormone in vitro. The complex thus formed expresses sequence-specific DNA-binding activity indistinguishable from that observed in the presence of hormone.

Animals↗

Studies in owl monkeys leading to the development of a synthetic vaccine against the asexual blood stages of Plasmodium falciparum.

During the development of a synthetic vaccine for human use against the asexual blood stages of Plasmodium falciparum, monkey trials were performed to assess safety, immunogenicity, and protectivity. We determined the minimal infective dose of the P. falciparum FVO strain, the kinetics of the immune response induced by vaccination with the synthetic peptide mixture (S7 + S12 + S17) or the synthetic hybrid polymeric protein SPf66, and the induction of protective immunity against the experimental challenge with 2 P. falciparum strains. A clear boosting effect was observed, determined by the increased antibody titers against synthetic peptides S7, S12, S17, and SPf66, and by improvement in the protective immune response against the challenge. These studies suggest that either the peptide mixture or the synthetic hybrid polymeric protein are excellent choices for the development of a vaccine against P. falciparum.

Amino Acid Sequence↗

Continuous warm blood cardioplegia: a new technique for myocardial protection.

Hypothermia is used to prolong the safe period of ischemic arrest by reducing the heart's oxygen demands. Due to this effect, hypothermia has been the fundamental component of most methods of myocardial protection during cardiac surgery. However, hypothermia has a number of unwanted side effects, such as detrimental effects on enzyme function, energy generation, and cell membranes. Since electromechanical arrest accounts for 90% of myocardial oxygen consumption, arresting the heart with chemical cardioplegia will reduce O2 consumption dramatically. Therefore, if the resting (arrested) heart is continuously perfused with oxygenated blood cardioplegia, one can easily provide the remaining 10% of O2 that it requires. Under these conditions, the need for hypothermia becomes questionable. In this paper, we describe the perfusionist's experience using the antegrade and retrograde technique of continuous warm blood cardioplegia.

Blood↗

High performance isoelectric focusing using capillary electrophoresis instrumentation.

High performance isoelectric focusing in capillaries provides rapid, high resolution separation of proteins based on their isoelectric points. Results can be obtained in a matter of minutes with little or no sample preparation. The technique requires the use of coated capillaries to reduce electroendosmosis so that stable, focused zones can be attained. Once focused, protein zones may be mobilized by the addition of salt to the catholyte or anolyte buffer. On-tube UV monitoring enables direct detection of sample components during mobilization with mass sensitivity equal to that of silver staining. The linear relationship between mobilization time and isoelectric point allows the technique to be used for estimation of protein pI. Demonstrated applications include separation of proteins in biological fluids with possible clinical applications, and characterization of biopharmaceutical proteins and monoclonal antibodies.

Capillary Action↗

Synaptosomal (Ca2+-Mg2+)-ATPASE activity modulation by cyclic AMP.

Dibutyryl cyclic AMP, in a concentration-dependent manner, increased synaptosomal (Ca2+-Mg2+)-ATPase activity, but in synaptic plasma membranes lacked any effect. The maximal enzyme activity in synaptosomes was increased by 38%, leaving unaltered the extrasynaptosomal Ca2+ concentration necessary to reach it. In the presence of 5 microM cyclic AMP, cyclic AMP-dependent protein kinase increased (30%) maximal (Ca2+-Mg2+)-ATPase activity in synaptic plasma membranes, but the apparent affinity for Ca2+ was not modified. This effect was partially inhibited (60%) by a cyclic AMP-dependent protein kinase inhibitor. The data suggest that synaptosomal (Ca2+-Mg2+)-ATPase activity is modulated by a cyclic AMP-dependent phosphorylation reaction.

Animals↗

Development of splenic natural suppressor (NS) cells in Ehrlich tumor-bearing mice.

Spleen cells from C57BL/6J mice bearing Ehrlich carcinoma growing as a solid tumor show progressive unresponsiveness to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogens. This is accompanied by striking spleen enlargement with marked hematopoietic activity. Lymphoproliferative assays of normal spleen cells in co-culture with tumor-bearing spleen cells (TBSC) show that: (a) TBSC contain non-specific suppressor cells able to abrogate both Con A and LPS responses, or mixed lymphocyte reaction, of normal spleen cells and (b) suppression by TBSC is MHC-unrestricted, non-prostaglandin-mediated and greatly enhanced by Con A supernatants. Suppressor cells associated with TBSC are large, low-density cells without markers of mature B or T lymphocytes or of the mononuclear phagocyte system. Most appear to be asialo-GM1-negative, as suppression was only partially inhibited by treatment with anti-asialo-GM1 and complement. Since NK activity is lacking in TBSC, our data strongly suggest that these "null" suppressor cells are related to the natural suppressor (NS) cells found described in normal bone-marrow and neonatal spleens, or induced in adult spleens by total lymphoid irradiation, graft-vs.-host disease, or cyclophosphamide treatment.

Animals↗

Inhibition of vascular endothelial cell prostacyclin synthesis by plasmin.

Vascular endothelial cells (EC) play an active role in the synthesis and assembly of components of the fibrinolytic system and the generation of the major fibrinolytic protease plasmin. However, the reciprocal effects of plasmin on EC function have not been previously examined. We have studied the actions of plasmin on the production of prostacyclin (PGI2) by cultured human umbilical vein (HUVEC) and bovine aortic (BAEC) endothelial cells. Plasmin causes little or no direct stimulation of PGI2 formation by EC. Preincubation of EC with plasmin, however, produces a time- and concentration-dependent inhibition of ionophore A23187-, thrombin-, and histamine-induced PGI2 synthesis; a smaller inhibitory effect on arachidonate- and PGH2-induced PGI2 synthesis is found. Incubation of HUVEC or BAEC with a physiologic concentration of plasminogen (180 micrograms/mL) and recombinant tissue plasminogen activator (tPA) generates tPA dose-dependent plasmin activity that exceeds that generated in the absence of EC. In the presence of plasminogen, tPA also causes a tPA dose-dependent inhibition of thrombin- and ionophore A23187-stimulated PGI2 production. PGI2 inhibitory plasmin activity is generated within the concentration range of tPA achieved in plasma during pharmacologic therapy with tPA. These findings suggest that vascular endothelial cells not only regulate activation of the fibrinolytic system but may also be targets of plasmin action on PGI2 synthesis in the modulation of hemostasis and thrombosis.

Animals↗

Doppler echocardiographic evaluation of right and left ventricular diastolic function in normal neonates.

Doppler echocardiograms of the tricuspid and mitral valves were recorded along with the electrocardiogram and respiration in 22 normal full-term neonates. A computer-interfaced digitizer pad was utilized to measure the following: peak E and A velocities (cm/s); E and A areas (the components of the total velocity-time integral in the early passive period of ventricular filling [E] and the late active period of atrial emptying [A], respectively) and the 1/3 area fraction (or the proportion of filling in the first 1/3 of diastole). All of the variables of right (tricuspid) versus left (mitral) ventricular filling were significantly different on the 1st day of life. Respective values were peak E velocity (cm/s) 44.6 +/- 10.0 (tricuspid) versus 53.2 +/- 9.3 (mitral), p less than 0.01; peak E/A ratio 0.84 +/- 0.14 versus 1.15 +/- 0.17, p less than 0.0001; E/total area 0.58 +/- 0.07 versus 0.63 +/- 0.05, p less than 0.005; E/A area ratio 1.05 +/- 0.23 versus 1.63 +/- 0.40, p less than 0.0001; 1/3 area fraction 0.31 +/- 0.04 versus 0.41 +/- 0.04, p less than 0.0001; peak A velocity (cm/s) 53.0 +/- 8.4 versus 47.6 +/- 5.8, p less than 0.05 and A/total area 0.57 +/- 0.09 versus 0.41 +/- 0.09, p less than 0.001; the mean heart rate (beats/min) was not significantly different: 121 +/- 8 versus 120 +/- 7. Most of the variables remained significantly different on the 2nd day of life, but the level of significance was the same or less for all measurements.(ABSTRACT TRUNCATED AT 250 WORDS)

Cesarean Section↗

Central regulation of gastric acid secretion by platelet-activating factor in anesthesized rats.

PAF has been implicated in the pathogenesis of acute gastric injury. When given peripherally, PAF induces severe gastric mucosal damage. PAF metabolizing enzymes are present in the brain but the central effects of PAF on the stomach are unknown. We have investigated in the rat the gastric secretion and gross mucosal integrity in response to intracerebroventricular (icv) PAF and compared it with that to icv TRH, a known central gastric secretagogue. Gastric acid output was markedly increased by TRH (171.6 +/- 26.3 mumol/h mean +/- SE) and by 20 micrograms/kg/h iv pentagastrin (107.6 +/- 23.6) when compared to controls receiving icv vehicle (20.2 +/- 7.5; p less than 0.01 for both). In contrast, acid output decreased after icv PAF (13.5 +/- 7.5). Furthermore, icv PAF markedly inhibited acid output stimulated by iv pentagastrin (45.1 +/- 7.03; p less than 0.05). Morphological studies showed acute gastric mucosal erosions after icv TRH and no damage was observed after icv PAF or vehicle. Thus, icv PAF reduces pentagastrin stimulated acid output and does not alter gastric mucosal integrity, whereas icv TRH stimulates acid secretion and induces gastric injury. The opposite effects of PAF and TRH suggests the existence of a gastric modulatory system at the central level.

Animals↗

Analysis of synaptic events in the opossum piriform cortex with improved current source-density techniques.

1. The piriform cortex of the opossum was studied by current source-density (CSD) analysis of field potentials to determine the laminar and temporal distribution of synaptic currents evoked by lateral olfactory tract (LOT) stimulation. 2. Extracellular conductivity was measured as a function of depth at high resolution and incorporated into CSD computations. Inclusion of the conductivity term resulted in relatively subtle changes in the shapes of CSD profiles. Resolution and accuracy of CSD computations was further improved by use of a new smoothing approach and averaging of multiple potential profiles obtained at the same site. 3. The CSD depth profile resulting from LOT stimulation revealed six major synaptic events that were consistently present at anterior, middle, and posterior sites: one during the first (A1) peak of the initial surface negative dichrotic field potential component, three during the second (B1) peak, one during the surface positive field potential component (period 2), and one during the second surface negative component (period 3). In addition, CSD profiles were computed for the population spike generated by synchronous discharge of action potentials. Depths of the net inward and outward membrane currents underlying these events were correlated with the cortical lamination as determined histologically by placement of small dye marks. 4. In agreement with previous reports it is concluded that the large inward membrane current in layer Ia during the A1 wave underlies a monosynaptic EPSP evoked in distal apical dendritic segments of pyramidal cells by afferent fibers. This EPSP displays a marked paired shock facilitation. 5. Based on anatomic and physiological considerations it is concluded that the three spatially and temporally distinct inward membrane currents (sinks) that were observed in layers III, superficial Ib, and mid- to deep-Ib during the B1 wave, underlie disynaptic EPSPs resulting from direct synaptic interactions between pyramidal cells. It is postulated that the layer III sink is generated in basal dendrites largely via local axon collaterals, the superficial layer Ib sink in intermediate apical dendritic segments by association fibers originating in the anterior piriform cortex, and the deep Ib sink in proximal apical segments by association fibers originating largely in the posterior piriform cortex. 6. The latencies of the layer Ia and superficial layer Ib sinks (presumed mono- and large disynaptic EPSPs, respectively) increased from anterior to posterior. Amplitude of the superficial Ib sink relative to the Ia sink increased from anterior to posterior.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Relation between delay and survival in 596 patients with breast cancer.

To evaluate the influence of delay between first symptom and first treatment upon survival the medical records of 596 patients with breast cancer were reviewed. The following intervals were considered: less than 3 months; 3-6 months and greater than 6 months. Patients in the less than 3 months delay group had a better distribution by clinical stages and a 10-year survival rate higher than those in the longer delay groups (p = 0.034). However, within each stage no statistically significant difference in survival according to delay was observed. A Cox multivariate analysis revealed that performance status and stage of disease were independent predictors of survival, but not delay. Assuming the best prognosis for patients with clinical stages I and II and less than 3 months delay, the group with longer delay times had 15 deaths over what would have been predicted. This adverse effect was observed almost exclusively among patients over age 50 (14/15).

Adult↗

Hormone-induced changes in the in vitro DNA-binding activity of the chicken progesterone receptor.

Previous analyses have indicated that steroid hormone receptors undergo an allosteric change in structure upon binding by the steroid ligand. This structural change was envisioned as an intramolecular unmasking of the protein's DNA-binding domain, thus allowing the receptor to function in gene regulation. We report an analysis of the effect of hormone on the DNA-binding activity of the chicken progesterone receptor. Using an isocratic elution of DNA affinity columns we show that unliganded receptor (aporeceptor) can bind a 23-basepair progesterone response element with high affinity and a high degree of sequence preference. Hormone causes a 1.5-fold increase in affinity for the PRE sequence and a 2-fold decrease in affinity for non-specific DNA. Kinetic analysis of the off-rate of receptor-DNA complexes is consistent with this minor effect of hormone. In addition, gel retardation analysis of receptor-progesterone response element complexes further substantiates that hormone is not required for sequence-specific DNA binding. These results indicate that hormone is not necessary for the progesterone receptor to fold into a conformation that recognizes specific gene regulatory sequences.

Animals↗

Histamine stimulated synaptosomal Ca2+ uptake through activation of calcium channels.

Histamine stimulated Ca2+ uptake in synaptosomes was completely inhibited by the slow Ca2+ channel antagonists verapamil, cinnarizine and flunarizine, and slightly inhibited by nifedipine and diltiazem. Ca2+ uptake in synaptosomes depolarized or predepolarized with varying K+ concentrations was increased by histamine, in both conditions, until 30mM K+. At higher K+ concentrations histamine was not able to alter K+ effects in either conditions. 30mM K+ stimulated uptake of Ca2+ in the absence or presence of histamine was not inhibited by verapamil and diltiazem. However nifedipine slightly inhibited K+ and K+ +histamine effects. 3-Isobutyl-1-methyl-xanthine and dibutyryl cyclicAMP potentiated (10%) the uptake of Ca2+ in synaptosomes induced by histamine. Dibutyryl cyclicAMP alone however decreased the basal Ca2+ uptake in a concentration-dependent manner. Verapamil, but not diltiazem, antagonized the effects elicited by 3-isobutyl-1-methyl-xanthine and dibutyryl cyclicAMP in the presence of histamine. The data suggest that the increase in synaptosomal Ca2+ uptake induced by histamine is mediated by the activation of the voltage sensitive calcium channels, and possibly a cyclicAMP-dependent protein kinase phosphorylation can modulate the opening of Ca2+ channels.

1-Methyl-3-isobutylxanthine↗

Histamine H2-receptor mediated activation of neonatal rat brain ornithine decarboxylase in vivo.

The effect of histamine (HA) administered via intracerebroventricular injection on ornithine decarboxylase (ODC) activity was studied in neonatal rat brain. The HA effect was dose and time dependent. Maximal increase in ODC activity was achieved 2 hr after administration of 10 micrograms HA (38% over control levels). Impromidine (HA H2-agonist) mimicked the effect of HA on ODC and ranitidine (HA H2-antagonist) inhibited the response to HA. Neither 2-thiazolylethylamine (HA H1-agonist) nor mepyramine (HA H1-antagonist) modified control ODC activity. The HA-releasers, compound 48/80 and polymixin B sulfate, elicited an increase in brain ODC activity of 35% and 32%, respectively, over the control value.

Animals↗

Clinical results with the TCu340 IUD.

A trial was carried out on the TCu340 intrauterine contraceptive device. The IUD was used by 431 women over a period of 2 years. The reasons for IUD removal were analyzed after 1 and 2 years using the life-table method. Later, the authors compared these results with the MLCu375 and the Nova-T, using the coefficient Z of Cramer. The authors concluded that the TCu340 was a high-load IUD with high effectiveness. However, there was a higher incidence of side-effects, such as pain, bleeding and expulsion in comparison to MLCu375.

Adolescent↗

Tuberculosis of the rectum in a patient with acquired immune deficiency syndrome. Report of a case.

Tuberculosis of the rectum is a rare disease. A patient with a miliary pattern of pulmonary tuberculosis had a rectal lesion which proved to be tuberculosis. The patient subsequently developed several opportunistic infections characteristic of acquired immune deficiency syndrome. The clinical, endoscopic, radiologic, and histologic findings of this treatable lesion are presented.

Acquired Immunodeficiency Syndrome↗