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E Martinez

Publications and source records attributed to E Martinez.

At least 199 records · Page 11Linked to original sources

The estrogen-responsive element as an inducible enhancer: DNA sequence requirements and conversion to a glucocorticoid-responsive element.

The estrogen-responsive element (ERE) present in the 5'-flanking region of the Xenopus laevis vitellogenin (vit) gene B1 has been characterized by transient expression analysis of chimeric vit-tk-CAT (chloramphenicol acetyltransferase) gene constructs transfected into the human estrogen-responsive MCF-7 cell line. The vit B1 ERE behaves like an inducible enhancer, since it is able to confer estrogen inducibility to the heterologous HSV thymidine kinase (tk) promoter in a relative position- and orientation-independent manner. In this assay, the minimal B1 ERE is 33 bp long and consists of two 13 bp imperfect palindromic elements both of which are required for the enhancer activity. A third imperfect palindromic element is present further upstream within the 5'-flanking region of the gene but is unable to confer hormone responsiveness by itself. Similarly, neither element forming the B1 ERE can alone confer estrogen inducibility to the tk promoter. However, in combinations of two, all three imperfect palindromes can act cooperatively to form a functional ERE. In contrast a single 13 bp perfect palindromic element, GGTCACTGTGACC, such as the one found upstream of the vit gene A2, is itself sufficient to act as a fully active ERE. Single point mutations within this element abolish estrogen inducibility, while a defined combination of two mutations converts this ERE into a glucocorticoid-responsive element.

Acetyltransferases↗

Cerebral phosphoinositide, triacylglycerol and energy metabolism during sustained seizures induced by bicuculline.

In ventilated rats, levels of phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol 4,5-bisphosphate (PIP2), diacylglycerol (DAG), triacylglycerol (TAG), free fatty acids (FFA) and phosphatidic acid, as well as their fatty acid contents, were measured in forebrain tissue after 1, 20 and 60 min of seizures induced by bicuculline. Cerebral energy state was also measured. PI decreased progressively throughout 60 min of seizures, whereas the levels of PIP and PIP2 did not change. DAG increased modestly and persistently. FFA increased markedly during the early seizure period, but decreased later. Following an initial drop, TAG rose above control. Phosphatidic acid did not change. The levels of ATP and energy charge potential decreased slightly and lactate accumulated. Stearic acid (18:0) and arachidonic acid (20:4) primarily accounted for the changes in the levels of the lipids. At the onset of seizures, the decrease of 18.0 and 20:4 in PI occurred in parallel with an enrichment of these fatty acids in FFA and DAG. Despite the fact that the losses of 18:0 and 20:4 from PI were quantitatively similar to each other at all times examined, the increase in free 18:0 was much larger than the increase in free 20:4 at 20 min of seizures. Concurrently there was a rise of 20:4 in TAG. As the FFA levels declined thereafter, 20:4 and docosahexaenoate (22:6) in TAG continued to increase. The results are consistent with the view that seizure activity stimulates the hydrolytic breakdown of brain phosphoinositides--the pathway catalyzed by phosphodiesterase of the phospholipase C type followed by lipases, and probably the pathway catabolized by phospholipases A as well. Preferential incorporation of polyunsaturated fatty acids into TAG-acyl residues may represent a mechanism to reduce the level of their free forms when the latter are produced in large amounts.

Animals↗

Electron microscopic visualization of protein-DNA interactions at the estrogen responsive element and in the first intron of the Xenopus laevis vitellogenin gene.

Stable protein-DNA complexes can be assembled in vitro at the 5' end of Xenopus laevis vitellogenin genes using extracts of nuclei from estrogen-induced frog liver and visualized by electron microscopy. Complexes at the three following sites can be identified on the gene B2: the transcription initiation site, the estrogen responsive element (ERE) and in the first intron. The complex at the transcription initiation site is stabilized by dinucleotides and thus represents a ternary transcription complex. The formation of the complexes at the two other sites is enhanced by estrogen and is reduced by tamoxifen, an antagonist of estrogen, while this latter effect is reversed by adding an excess of hormone. No sequence homology is apparent between the site containing the ERE and the binding site in intron I and functional tests in MCF-7 cells suggest that these two sites are not equivalent. Finally, we made use of previously characterized deletion mutants of the 5' flanking region of the gene B1, a close relative of the gene B2, to demonstrate that the 13-bp palindromic core element of the ERE is involved in the formation of the complexes observed upstream of the transcription initiation site.

Animals↗

Cerebral phosphoinositide and energy metabolism during and after insulin-induced hypoglycemia.

During and after insulin-induced hypoglycemia, changes in levels of cerebral phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidic acid (PA), triacylglycerol (TAG), diacylglycerol (DAG), and free fatty acids (FFAs) as well as the cerebral energy state were studied in relation to the EEG. In hypoglycemic rats with an EEG pattern of quasiperiodic sharp or slow sharp waves, which preceded the development of an isoelectric EEG, PIP2 levels increased significantly, together with a slight decrease in PI content. Levels of the other lipids did not change during this period. The cerebral energy state was affected only slightly in spite of profound decreases in plasma and tissue glucose levels. With 30 min of an isoelectric EEG, levels of all phosphoinositides and PA decreased significantly; total FFA and DAG contents increased seven- and twofold, respectively; the TAG-palmitate level decreased, and that of TAG-arachidonate increased. Plasma and tissue glucose were nearly depleted, and the cerebral energy state deteriorated severely. The increment in fatty acids in the DAG and FFA pools was less than their loss from phosphoinositides and PA, an observation suggesting vascular washout or oxidation of a portion of the FFAs produced. Following 90 min of glucose infusion, PIP and PA levels recovered to control values; however, the PIP2 content exceeded control levels, and that of PI remained below control levels. DAG and FFA contents returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

"Results of large surface hemodialysis (LSH): a shorter and cheaper treatment".

A surface area of 5 m2, a QB of 500 ml/min and a QD of 1000 ml/min reduces the time of hemodialysis to just 6 hours per week. All the patients had been in a conventional dialysis program for at least 2 years. These patients have now been in LSH for a period of between 2 and 3 years. Most patients have no appreciable residual function, they follow a free diet, living a normal life, and weigh between 51 and 88 Kg, ranging in ages from between 26 and 69 years. We use this technique with 3 schedules: Schedule I) 2 hours/3 days/week; Schedule II) 3 hours/2 days/week; Schedule III) 6 hours/1 day/week. In the 3 schedules the clinical and biochemical results have been the same as with conventional dialysis. With this technique the patients have greater freedom and comfort because they avoid 312 hours per year connected to the artificial kidney. The patients in Schedule II also avoid 52 dialysis sessions and 104 trips to the dialysis center per year. And the patients in Schedule III also avoid 104 dialysis sessions and 208 trips to the center per year. Furthermore, the cost savings in Schedule II is $2,500 per year/patient and in Schedule III $8,500 per patient/year.

Adult↗

Acute free colonic perforation as a presenting sign of Crohn's disease. Case report.

A case of free perforation of the colon in Crohn's disease is reported. There was no dilation or distal obstruction of the gut. Apart from the perforation, no macroscopic (transmural) affection of the gut was found at laparotomy. The diagnosis was not achieved until the follow-up period. The incidence and pathogenesis of the complication are discussed.

Adult↗

Glomerular function and morphology after renal mass reduction in dogs.

To determine whether the proteinuria, glomerular sclerosis, and decline in glomerular filtration rate (GFR) described in rodents after renal mass reduction develop in another species, 24-hour proteinuria, glomerular structure, and fasting and postfeeding GFR were examined in dogs subjected to seven-eighths reduction in renal mass. All dogs were fed a diet containing 26% protein. Six dogs with a GFR less than 10 ml/min (8% to 17% of control two-kidney GFR) were killed within 6 months after renal mass reduction. Twenty-four-hour urinary protein excretion was modestly although definitely increased (236 +/- 26 mg/24 hr, P less than 0.01). All remnant kidneys demonstrated structural changes of mesangial hyperplasia or focal glomerular sclerosis. Ten dogs with a remnant kidney and early GFRs 16% to 39% of control values were followed for 18 to 39 months. In seven dogs, GFR showed little tendency to decrease with time. In one of them, proteinuria was 106 mg/24 hr with normal-appearing glomeruli at 14 months. In three dogs, proteinuria was progressive, averaging about 1 gm/24 hr at 18 months and 2 gm/24 hr at 24 to 34 months; glomerular pathologic findings progressed from focal mesangial hyperplasia or focal glomerular sclerosis at 8 to 16 months to focal and segmental sclerosis or diffuse glomerular obsolescence at 25 to 34 months; and fasting GFR progressively declined starting at 21 to 24 months after renal mass reduction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Identification of estrogen-responsive DNA sequences by transient expression experiments in a human breast cancer cell line.

The expression of a hybrid gene formed by the promoter region of the Xenopus laevis vitellogenin gene B1 and the CAT coding region is regulated by estrogen when the gene is transfected into hormone-responsive MCF-7 cells. Furthermore, the 5' flanking region of the gene B1 alone can confer inducibility to heterologous promoters, although to a varying extent depending on the promoter used. Deletion mapping of he vitellogenin hormone-responsive sequences revealed that a 13 bp element 5'-AGTCACTGTGACC-3' at position -334 is essential for estrogen inducibility. We have shown previously that this 13 bp element is present upstream of several liver-specific estrogen-inducible genes.

Acetyltransferases↗

Diagnosis of fetal arrhythmias using echocardiographic and Doppler techniques.

Fetal echocardiography is the most practical method for diagnosing prenatal arrhythmias. Because some prenatal tachyarrhythmias have been shown to respond to antiarrhythmic drugs, correctly diagnosing fetal arrhythmias has assumed new importance. With the aid of two-dimensional echocardiographic imaging, an M-mode cursor can be aligned to record atrial and ventricular wall motion--either independently or simultaneously. A consistent feature in the fetus is prominent atrial wall contractions that can be readily recorded on the M-mode tracing. By matching atrial and ventricular wall contractions with assumed P waves and QRS complexes, the fetal electrocardiogram can be reconstructed. In 57 fetuses studied, recurrent atrial and ventricular ectopic beats were the most common prenatal arrhythmias. However, atrial flutter, ventricular tachycardia, atrial and ventricular bigeminy and atrial and ventricular bradyarrhythmias have been correctly identified and in some instances appropriately treated. Marked fetal bradycardia in the midtrimester of pregnancy is shown for the first time to be caused by transducer pressure on the maternal abdominal wall.

Arrhythmias, Cardiac↗

Cerebral phosphoinositide, triacylglycerol, and energy metabolism in reversible ischemia: origin and fate of free fatty acids.

Levels of phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol (PI), phosphatidic acid, diacylglycerol (DAG), triacylglycerol (TAG), and free fatty acids (FFAs), as well as their fatty acid composition, were determined in rat forebrain during ischemia and postischemic recirculation. Cerebral energy state and electroencephalograms (EEGs) were also studied. Fifteen minutes of ischemia resulted in a decrease in PIP2 and PIP contents but not in PI content, concurrent with an enlargement of the FFA and DAG pools. The latter were enriched in stearate and arachidonate. Prolongation of ischemia did not produce further changes in content of any of the inositol phospholipids, but the increase in levels of FFAs and DAG continued. At the end of 45 min of ischemia, levels of both PIP2 and PIP decreased by 45-50%, and the total phosphoinositide content (PIP2 + PIP + PI) decreased by 21%, whereas levels of FFAs and DAG increased to 14- and 3.6-fold of control levels, respectively. During ischemia, the TAG-palmitate level decreased, but the TAG-arachidonate level increased; the tissue energy state deteriorated severely; and the EEG was suppressed. A 30-min recirculation period after 15 or 45 min of ischemia led to increases in PIP2, PIP, and total phosphoinositide contents, whereas levels of FFAs and DAG promptly decreased toward control values. The TAG-arachidonate level peaked and the TAG-palmitate level returned to a low control value during early recirculation. The ischemic changes in tissue lipids were completely reversed within 3 h of recirculation after both periods of ischemia. Adenylates were fully phosphorylated with as little as 30 min of reflow. The EEG activity partially recovered during reflow after 15 min of ischemia, whereas it remained depressed after prolonged ischemia. Thus, phosphodiesteric cleavage of PIP2 and PIP followed by deacylation of DAG is likely to contribute to the production of FFAs in early ischemia. Deacylation of undetermined lipids plays a role for the increment in levels of FFAs in the later period of ischemia. The rapid postischemic increase in levels of PIP2 and PIP indicates active synthesis not only from existing PI, but probably also by means of accumulated FFAs and DAG. These results indicate that the impaired resynthesis of inositol phospholipids cannot be a cause of the poor EEG activity after prolonged ischemia. Degradation and resynthesis of polyphosphoinositides and formation of TAG-arachidonate may be important for modulation of free arachidonic acid levels in the brain during temporary ischemia.

Animals↗

Large-surface hemodialysis.

A hemodialysis device with a surface of 5 m2, a blood flow (QB) of 500 ml/min, and a dialysate flow (QD) of 1,000 ml/min has enabled the authors to obtain in 6 h/week the same clearances for urea, creatinine, uric acid, phosphates, and vitamin B12 as has conventional hemodialysis (CH), which takes from 12 to 15 h/week. Twenty-five patients were hemodialyzed throughout 1 year with CH and another year with large-surface hemodialysis, 20 with a creatinine clearance (CCr) of 0.5 ml/min and 5 with a CCr between 0.5 and 4.5 ml/min. All followed a free diet and led a normal life. Hemodialysis time was 6 h/week, and the results obtained were equal to those of the previous year with a CH of 12 h/week. The use of a bicarbonate dialysate rich in glucose, with a relatively high level of potassium and sodium, can avoid the disequilibrium syndrome caused by quick hemodialysis and makes possible the removal of excess liquid in only 6 h/week, without disturbances for the patient.

Adult↗

Assignment of symbiotic developmental phenotypes to common and specific nodulation (nod) genetic loci of Rhizobium meliloti.

Rhizobium meliloti nodulation (nod) genes required for specific infection and nodulation of alfalfa have been cloned. Transposon Tn5 mutagenesis defined three nod regions spanning 16 kilobases of the pSym megaplasmid. Genetic and cytological studies of 62 nodulation-defective mutants allowed the assignment of symbiotic developmental phenotypes to common and specific nod loci. Root hair curling was determined by both common (region I) and specific (region III) nod transcription units; locus IIIb (nodH gene) positively controlled curling on the homologous host alfalfa, whereas loci IIIa (nodFE) and IIIb (nodH) negatively controlled curling on heterologous hosts. Region I (nodABC) was required for bacterial penetration and infection thread initiation in shepherd's crooks, and the nodFE transcription unit controlled infection thread development within the alfalfa root hair. In contrast, induction of nodule organogenesis, which can be triggered from a distance, seemed to be controlled by common nodABC genes and not to require specific nod genes nodFE and nodH. Region II affected the efficiency of hair curling and infection thread formation.

Cloning, Molecular↗

Effects of beta-blockade on exercise capacity of trained and untrained men: a hemodynamic comparison.

To study the effects of cardiovascular fitness on hemodynamic responses to exercise during beta-adrenergic blockade (BAB), submaximal [60% of maximum O2 uptake (VO2max)] and maximal treadmill exercise data were collected in 11 trained (T, VO2max 63.3 ml X kg-1 X min-1, 26.8 yr) and 11 untrained (UT, VO2max 44.5 ml X kg-1 X min-1, 25.0 yr) male subjects. Subjects completed two maximal control tests followed by a randomized, double-blind series of maximal tests after 1-wk treatments with placebo (PLAC), propranolol (PROP, 160 mg/day, beta 1- and beta 2-blockade), and atenolol (ATEN, 100 mg/day, beta 1-blockade). Treatments were separated by 1-wk washout periods. At 60% of control VO2max T and UT subjects experienced no reductions in O2 uptake (VO2) with either drug. Submaximal heart rate (HR, beats/min) was 134.8 PLAC, 107.0 PROP, 107.9 ATEN (P less than 0.05 both drugs vs. PLAC) in T subjects and 141.1 PLAC, 106.1 PROP, and 105.0 ATEN (P less than 0.05 both drugs vs. PLAC) in UT subjects. Cardiac output (1/min) for T was 17.3 PLAC, 16.9 PROP, 16.5 ATEN (P less than 0.05 ATEN vs. PLAC in T only) and for UT it was 12.2 (PLAC), 11.7 (PROP), 11.5 (ATEN) (P less than 0.05 both drugs vs. PLAC in UT). Stroke volume increased from 129.8 ml (PLAC) to 158.6 (PROP) and 156.2 (ATEN) in T (P less than 0.05 both drugs vs. PLAC) and from 86.8 (PLAC) to 110.0 (PROP) and 109.8 (ATEN) (P less than 0.05 both drugs vs. PLAC) in UT. The increases in stroke volume (SV) were similar in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗