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Biomedical subjects

J M Pena

Publications and source records attributed to J M Pena.

27 records · Page 2Linked to original sources
PubMed1983

Cyclic adenosine 3':5'-monophosphate-dependent and -independent protein kinases in human leukemic cells.

The pattern of protein kinase activity in leukemic cells from patients with chronic myelocytic leukemia, acute myeloblastic leukemia, acute monocytic leukemia, chronic lymphocytic leukemia, and acute lymphoblastic leukemia was studied and compared with normal peripheral blood granulocytes and lymphocytes. Our data showed that: (a) histone kinase activity was slightly lower in leukemic cells than in normal cells, whereas casein kinase activity was 2- to 3-fold higher in leukemic cells; (b) cyclic adenosine 3':5'-monophosphate stimulated 1.4- to 1.6-fold histone kinase activity of both normal and leukemic cells, whereas it did not stimulate casein kinase activity; (c) the ratio of histone kinase activities to casein kinase activities correlated directly with the maturation of the white blood cells; and (d) histone and casein kinase activities of extracts from normal and leukemic cells behaved similarly on chromatography on phosphocellulose and casein/Sepharose 4B. These results suggest that the increase in casein kinase activity is not due to the appearance of a new type of casein kinase but to an increase of the casein kinases 1 and 2 present in normal cells.

Casein Kinases↗
PubMed1983

Intralipid and free plasmatic tryptophan in vitro.

In an attempt to investigate the role of the lipidic emulsion Intralipid in the development of metabolic encephalopathy in a patient showing high free tryptophan levels, the relationship between lipidic emulsion and free tryptophan was examined in in vitro experiments. The addition of intralipid to normal serum produces an immediate increase in non-esterified fatty acids and a parallel rise in free tryptophan. Moreover, when serum with intralipid is incubated at 37 degrees C, the lipases release new non-esterified fatty acids and the free tryptophan increases proportionally. The non-esterified fatty acid content of intralipid was found to be 12 +/- 2 mEq X 1(-1). An inverse correlation was seen between free tryptophan and different serum albumin concentrations. It is concluded that intralipid causes an increase in free tryptophan levels. It is known that in vivo free tryptophan modulates 5-hydroxytryptamine synthesis and thus may be considered a possible causal agent for encephalopathy.

Bilirubin↗
PubMed1981

Purification and characterization of two cyclic AMP-independent casein/glycogen synthase kinases from rat liver cytosol.

Two cyclic AMP-independent protein kinases (ATP: protein phosphotransferase, EC 2.7.1.37) (casein kinase 1 and 2) have been purified from rat liver cytosol by a method involving chromatography on phosphocellulose and casein-Sepharose 4B. Both kinases were essentially free of endogeneous protein substrates and capable of phosphorylating casein, phosvitin and I-form glycogen synthase, but were inactive on histone IIA, protamine and phosphorylase b. They were neither stimulated by cyclic AMP, Ca2+ and calmodulin, nor inhibited by the cyclic AMP-dependent protein kinase inhibitor protein. The casein and glycogen synthase kinase activities of each enzyme decreased at the same rate when incubated at 50 degrees C. Casein kinase 1 and casein kinase 2 showed differences in molecular weight, sensitivity to KCl, Km for casein and phosvitin and Ka for Mg2+, whereas their Km values for ATP and I-form glycogen synthase were similar. The phosphorylation of glycogen synthase by these kinases correlated with a decrease in the +/- glucose 6-phosphate activity ratio (independence ratio). However, casein kinase 1 catalyzed the incorporation of about 3.6 mol of 32P/85000 dalton subunit, decreasing the independence ratio from 83 to about 15, whereas the phosphorylation achieved by casein kinase 2 was only about 1.9 mol of 32P/850000 dalton subunit, decreasing the independence ratio to about 23. The independence ratio decrease was prevented by the presence of casein but was unaffected by phosphorylase b. These data indicate that casein/glycogen synthase kinases 1 and 2 are different from cyclic AMP-dependent protein kinase and phosphorylase kinase.

Animals↗
PubMed1981

Cyclic AMP-independent casein/glycogen synthase kinases from pig polymorphonuclear leucocytes.

1. Two cyclic AMP-independent casein/glycogen synthase kinases were purified from pig polymorphonuclear leucocytes by chromatography on phosphocellulose followed by affinity chromatography on casein-Sepharose 4B or gel filtration on Bio-Gel A-1.5m. When the affinity step was used, the specific activities were 86 and 43units/mg of protein for casein kinase 1 and 2, respectively, whereas these values were 94 and 90units/mg of protein when the gel-filtration step was used. 2. These kinases differ as follows: (a) the molecular weight of casein kinase 1 (38000) is very much lower than that of casein kinase 2 (185000); (b) the K(m) for casein (0.46mg/ml) and K(a) for Mg(2+) (0.3mm) of casein kinase 1 are lower than those of casein kinase 2 (0.90mg/ml and 1.7mm respectively); (c) KCl stimulates the phosphorylation of casein by casein kinase 1, whereas it inhibits phosvitin phosphorylation by this enzyme; on the contrary, the effect of KCl on casein kinase 2 is very similar with either casein or phosvitin as substrate; (d) although both kinases phosphorylate rabbit muscle glycogen synthase I, the ratio of glycogen synthase to casein phosphorylation by casein kinase 1 is about 4-fold greater than that by casein kinase 2. Furthermore, (32)P incorporation into glycogen synthase promoted by casein kinase 1 (3.6mol of (32)P/mol of 85000-dalton subunit) is twice that observed with casein kinase 2 (1.8mol of (32)P/mol of 85000-dalton subunit). Such a phosphorylation results in a decrease in the glucose 6-phosphate-independence ratio of glycogen synthase to 10-15 with casein kinase 1 and to 35-45 with casein kinase 2. 3. The activity of both kinases is neither stimulated by cyclic AMP, Ca(2+) and calmodulin nor inhibited by cyclic AMP-dependent protein kinase inhibitor protein. 4. No phosphorylation kinase activity was observed with casein kinase 1 and 2 at either pH6.8 or 8.2 in the presence of Ca(2+). 5. Activities of both kinases on casein and glycogen synthase decreased in parallel when incubated at 50 degrees C.

Animals↗
PubMedDate not supplied

Reagent strip for semiquantitative total and specific IgE measurement: a comparison to radioisotopic methods.

Determination of total and specific IgE using reactive test strips may afford a useful screening method in the in vitro diagnosis of allergens owing to easy use and possible correlation with established methods. T.D. LETI reactive strips for the semiquantitative determination of total IgE is a method combining classical fundamentals (sandwich anti-IgE-IgE-labeled anti-IgE) and the use of strip methods. It also permits determination of specific IgE levels in a wide range of Mediterranean and food (10) allergens. In a preliminary assessment, this method was correlated with isotopic PRIST for total IgE and with Phadebas RAST for specific IgE. The most important features are: a) no additional equipment besides serum or whole blood; b) cheaper and faster than PRIST for total IgE; c) correlation coefficient of 0.88 in the proposed range (n = 20) and 0.91 (n = 25) in the 10-500 U/ml range for PRIST/T.D. LETI for total IgE; d) specific IgE determination strips present sensitivity and specificity of 88 and 91%, respectively, in 180 allergens assayed in relation to Phadebas RAST. In conclusion, the in vitro diagnosis of allergens using reactive strips is currently a practical and simple method. By optimizing the top of the scale for total IgE and reducing the volume and/or time in specific IgE, it may prove to be an important screening method.

Allergens↗
PubMedDate not supplied

Specific IgE determination using the CAP system: comparative evaluation with RAST.

The CAP system's main contribution is its solid phase, which consists of a cellulose polymer activated within a capsule (the ImmunoCAP). This solid phase can bind more protein to it, and, in addition, the conditions of reaction seem to make the system more sensitive at detecting antibodies to certain antigens. It is therefore important to assess the new analytical and diagnostic performance in comparison with previous systems. In this context, we studied the reliability and comparison with the RAST and with skin tests carried out on 144 pediatric patients. Skin tests and specific IgE for radioimmunological RAST (radio-allergosorbent test) and for the fluoroimmunological CAP system were performed on all the patients. The RAST/CAP correlation quotients for the different allergens tested varied between 0.971 and 0.991. Diagnostic sensitivity increased for all the allergens studied and specificity remained unchanged. The system provides reliable results, with better diagnostic capacity than RAST, but it must be quantified for each allergen because its results are not interchangeable.

Adolescent↗
PubMedDate not supplied

Determination of total and specific IgE using UNICAP 100: comparative study with the CAP system.

The methods of serum determination of specific IgE to different allergens have showed lower diagnostic sensitivity than the alternative in vivo methods, the skin tests. The CAP system from Pharmacia, owing to its solid phase, has ameliorated this disadvantage, showing in various studies greater diagnostic sensitivity than the classic RAST, without affecting specificity. However, this system is still semi-automatic and requires daily calibration. UNICAP 100 is a completely automatic autoanalyser for total IgE, specific IgE and Eosinophil Cationic Protein, which combines the high sensitivity of the CAP system with complete automation and monthly calibration. The aim of the present study is to assess the practicality and reliability of UNICAP 100, when compared to the CAP System, for the determination of total IgE and specific IgE, as well as the sensitivity and diagnostic specificity; using as a reference skin tests in 150 paediatric patients. The coefficients of variation in the study of intraseries imprecision ranged between 2.1% and 3.6% for total IgE and between 2.2% and 5.1% for specific IgE, depending on the allergen and the level studied. The intraseries imprecision ranged between 3.3% and 7.7% for total IgE and between 5.2% and 8.9% for specific IgE. The coefficients of correlation of the study of interchangeability with the results of the CAP System varied between 0.985 and 0.998, all the allergens tested (9) being interchangeable. Finally, the diagnostic sensitivity varied between 70% and 95% and the specificity between 87% and 100%. In conclusion, UNICAP 100 showed results that were interchangeable with the CAP System, noticeably improving the benefits owing to its complete automation and its calibration system.

Allergens↗