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Biomedical subjects

J Arenas

Publications and source records attributed to J Arenas.

At least 181 records · Page 10Linked to original sources

Arylsulfatase A in urine of patients with urothelial tumors.

A significant increase in urine arylsulfatase A activity (p less than 0.01) was found in patients with urothelial tumors. Arylsulfatase A activity was 1.36 +/- 1.10 U/24-h urine in control specimens, 1.90 +/- 1.66 U/24-h urine in various genitourinary tract disorders, and 3.90 +/- 1.98 U/24-h urine in transitional cell carcinoma specimens. Surgical treatment of the neoplastic patients lowered the arylsulfatase A activity found in urine to within reference values. The arylsulfatase A excreted by patients with these tumors was highly sensitive to thermal inactivation while the enzyme activity in the control urines was less affected by the heat treatment. The time course of the arylsulfatase A reaction with 4-nitrocatechol sulfate was not linear in normal individuals, while it was linear in 90% of patients with urothelial tumors. This difference in the kinetic pattern of the enzyme could be used to increase the diagnostic specificity of the determination.

Adult↗

Activities of creatine kinase and its isoenzymes in serum in various skeletal muscle disorders.

We studied possible correlations between anatomopathological and clinical features and the values for total creatine kinase (CK; EC 2.7.3.2) and its isoenzymes, including the proportion of CK-MB, in a population displaying several neuromuscular pathologies. Although we observed no specific isoenzyme pattern associated with the different myopathies, we found isoenzyme analysis useful in studying the histopathological evolution of illness. We also considered whether the pathology was regenerative or nonregenerative, and what type of fiber (I or II) was involved. High CK-MB percentages (greater than 6%) were associated with regenerative and type I fiber myopathies, with regenerative type tissues being the principal factor associated with an increasing proportion of CK-MB. Studying the changes in CK-MB percentage in serum appears to be useful in discriminating neuromuscular from myocardial pathologies.

Adolescent↗

Total creatine kinase (CK) and CK-B activity in maternal blood and cord-blood samples after vaginal and cesarean births.

We studied variations in the activity of total creatine kinase (CK; EC 2.7.3.2) and of CK-B in maternal and cord-blood samples, comparing data obtained for vaginal and cesarean births. CK-B activity was determined with an immunoinhibition assay. In all cases, there was a significant postpartum increase in total CK and in CK-B activity in maternal sera, whereas cord-blood samples showed no significant differences between activities in arterial and venous blood for either vaginal or cesarean births. Statistically significant differences were found in CK-B activity, but not in total CK, between cord-blood samples from vaginal births and those from cesareans.

Cesarean Section↗

In vitro formation of a lariat structure containing a G2'-5'G linkage.

We have examined the effects of base changes at the lariat branch site of a modified adenovirus major late precursor mRNA (pre-mRNA). Replacement of the A residue at the lariat attachment site with a G residue was studied. Incubation of this altered pre-mRNA with nuclear extracts of HeLa cells yielded less spliced mRNA (10-fold) than similar reactions with the wild type pre-mRNA. The intron lariat formed during the reaction with the mutant transcript contained the predominant branch (2'-5' phosphodiester linkage) to an upstream A residue. In contrast, the intron/exon 2 lariat contained the predominant branch to the substituted G residue. These results indicated that detectable spliced RNA was formed when the lariat was attached at the A residue but not when the lariat was attached to the substituted G residue. A second mutation was introduced into the transcript by substituting an additional G residue at the alternative A branch site. When transcript derived from this plasmid was incubated with nuclear extract, cleavage occurred at the 5' splice site, and an intron/exon 2 lariat was produced, but spliced RNA was not detected. T1 RNase digestion and primer extension analyses of this intron/exon 2 lariat revealed that all of the lariat formed on the G residue at the normal attachment site.

Adenoviridae↗

Purification of a RNA debranching activity from HeLa cells.

The splicing of messenger RNA precursors (pre-mRNA) of eukaryotic cells involves the formation of a branched RNA intermediate known as a RNA lariat. This structure is formed in the first step of the reaction when a cleavage at the 5' splice site generates the 5' exon and a RNA species containing the intron and 3' exon in which the phosphate moiety at the 5' end of the intron is forming a 2'-5' phosphodiester bond with the 2'-hydroxyl moiety of a specific adenine residue near the 3' end of the intron forming a RNA branch with the following structure: -pA2'-pX-3'-pZ-. We have purified a debranching activity approximately 700-fold from the cytosolic fraction of HeLa cells. This activity catalyzes the hydrolysis of the 2'-5' phosphodiester bond of branched RNA structures yielding a 5'-phosphate end and a 2'-hydroxyl group at the branch attachment site. The activity possessed a sedimentation coefficient of 3.5 S. The reaction catalyzed by the purified fraction requires a divalent cation and is optimal at pH 7.0. The purified activity can efficiently hydrolyze triester trinucleotide structures (pY2'-pX-3'-pZ-) prepared by digestion of RNA lariats with nuclease P1. In contrast, a 2' phosphate monoester product (-pG2'-p 3'-pC-), formed by the wheat germ RNA ligase, was not attacked.

Cations, Divalent↗

Maternal cell-mediated sensitisation to paternal HLA may occur, but is not a regular event in normal human pregnancy.

Maternal cell-mediated sensitisation to paternally-inherited fetal HLA was studied in 10 primigravid and 9 multigravid women. One woman in each group was found to have circulating cytotoxic effector cells specific for paternal lymphocytes, but both had a normal pregnancy outcome. These results show that sensitisation is not a regular event in pregnancy and that there must therefore be a block in the afferent (recognition) phase of the mother's immune response. Where sensitisation does occur there must also be a block on the efferent (effector) arm of the response to prevent rejection of the fetus. The restricted distribution of major histocompatibility complex antigens on trophoblast and the function of the placenta as a barrier between maternal and fetal immune compartments can explain the occurrence of both types of blockade.

Cytotoxicity, Immunologic↗

Catalytic concentrations of amylase isoenzymes: an assay with wheat-germ inhibitor and 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate.

We coupled a kinetic procedure to a selective inhibiting method for determining amylase isoenzymes in biological samples, using 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate and a wheat-germ selective inhibitor with the Gilford S-III spectrophotometer. On plotting remaining amylase activities/total amylase activities (R/T) vs pancreatic amylase activities/salivary amylase activities (P/S) ratios, we found the curve to be linear for P/S ratios from 0.2 to 5. The inhibition rate of amylase inhibitor was constant in solutions having total amylase activities between 20 and (at least) 900 U/L. CVs were 3.1 to 7.1% for pancreatic amylase and 2.0 to 12.9% for salivary amylase in serum. Correlation with the Phadebas method was excellent (r = 0.99) for both pancreatic and salivary amylase. We also automated this procedure in an Hitachi 705 analyzer and correlated the results (r = 0.99) with those by our manual method.

Amylases↗

The enzymatic conversion of 3'-phosphate terminated RNA chains to 2',3'-cyclic phosphate derivatives.

The enzyme, RNA cyclase, has been purified from cell-free extracts of HeLa cells approximately 6000-fold. The enzyme catalyzes the conversion of 3'-phosphate ends of RNA chains to the 2',3'-cyclic phosphate derivative in the presence of ATP or adenosine 5'-(gamma-thio)triphosphate (ATP gamma S) and Mg2+. The formation of 1 mol of 2',3'-cyclic phosphate ends is associated with the disappearance of 1 mol of 3'-phosphate termini and the hydrolysis of 1 mol of ATP gamma S to AMP and thiopyrophosphate. No other nucleotides could substitute for ATP or ATP gamma S in the reaction. The reaction catalyzed by RNA cyclase was not reversible and exchange reactions between [32P]pyrophosphate and ATP were not detected. However, an enzyme-AMP intermediate could be identified that was hydrolyzed by the addition of inorganic pyrophosphate or 3'-phosphate terminated RNA chains but not by 3'-OH terminated chains or inorganic phosphate. 3'-[32P](Up)10Gp* could be converted to a form that yielded, (Formula: see text) after degradation with nuclease P1, by the addition of wheat germ RNA ligase, 5'-hydroxylpolynucleotide kinase, RNA cyclase, and ATP. This indicates that the RNA cyclase had catalyzed the formation of the 2',3'-cyclic phosphate derivative, the kinase had phosphorylated the 5'-hydroxyl end of the RNA, and the wheat germ RNA ligase had catalyzed the formation of a 3',5'-phosphodiester linkage concomitant with the conversion of the 2',3'-cyclic end to a 2'-phosphate terminated residue.

Adenosine Triphosphate↗

Isolation and characterization of two fractions from HeLa cells required for mRNA splicing in vitro.

A nuclear extract from HeLa cells has been separated by DEAE-cellulose chromatography into two fractions, both of which are required for mRNA splicing in vitro. Both fractions are heat labile and sensitive to N-ethylmaleimide. The activity of one of the fractions was abolished by preincubation with micrococcal nuclease, while the other fraction was unaffected by this treatment. This abolition indicates an essential nucleic acid component. Fractions I and II are required for the in vitro splicing of human beta-globin and adenovirus transcripts.

Chromatography, DEAE-Cellulose↗

[Epidemic outbreak of tinea corporis due to Microsporum gypseum].

A small epidemic of tinea corporis due to M. gypseum is reported. There were 13 children affected, ages 1-15 years. These children belonged to 6 neighbouring families and all used a common "playground", an empty lot located nearby. Cultures were positive for M. gypseum in the 13 children and the agent was also isolated from 2 soils collected in the playing-ground. Soil isolates were classified as N. gypsea. Clinically, the lesions were circinated and had active borders, they were preferentially located in the trunk. Most children (8/13) had multiple lesions. These and other pertinent aspects are discussed in the text.

Adolescent↗

Acetaminophen-induced fulminant hepatic failure in dogs.

Results concerning morphological and biochemical changes following intravenous administration of different doses of acetaminophen in dogs are reported. Acetaminophen infusion, as a parenteral solution (500 mg per kg per 90 min), produced fulminant hepatitis characterized by a good correlation between Portmann's grade of lesion and percentage of necrosis. All animals died within 76 hr after intoxication. Analysis of biochemical parameters revealed positive correlation between serum bilirubin levels and severity of the hepatic lesion. The experimental model of acetaminophen-induced hepatotoxicity is proposed as a model for evaluation of the therapeutic efficacy of new medical and surgical procedures.

Acetaminophen↗

An electrically powered total artificial heart. Over 1 year survival in the calf.

An electric motor driven orthotopic artificial heart was implanted in a 110 kg female Holstein calf as part of a series of 12 such implants intended to demonstrate the in vivo durability and compatibility of the device. The device uses pusher plates set into motion by a reversing brushless DC motor and roller screw to alternately eject two cylindrical sac type blood pumps. The pumps use Bjork-Shiley Delrindisc convexo-concave or monostrut valves. The left pump provides an 88-90 ml dynamic stroke volume. Woven Dacron grafts and polyurethane coated Dacron/Lycra cuffs are used to attach the device to the major arteries and atria, respectively. A polyurethane conduit and anchoring skin button bring motor wires percutaneously to an extracorporeal controller. The controller provides balanced cardiac output sensitive to atrial or aortic pressures, without operator intervention. The system is hermetically sealed and uses a simple compliance sac to maintain thoracic pressure between the pumps. The calf recovered uneventfully from surgery and thrived thereafter. She was killed on the 388th post-operative day because of worsening cardiac insufficiency. The previous three operative survivors in this series lived 131, 134, and 204 days. These results indicate the device's good potential for durability and body compatibility.

Animals↗