Anticuerpos monoclonales que se unen a la hormona del crecimiento humana (hGH)

Traducción de Patente Europea E97916700 (fecha de solicitud, 27/03/1997).-- Prioridad: SE199603299601231.-- Titulares: Pharmacia Spain S.A., Consejo Superior de Investigaciones Científicas (CSIC).La invención se refiere a anticuerpos monoclonales capaces de unirse específicamente a la variante de la hormona de crecimiento humana de peso molecular 20kda. Dicho anticuerpo monoclonal no presenta prácticamente unión a la hgh de 22 kda. La invención se refiere asimismo a la utilización de dicho anticuerpo monoclonal para la medición de la hgh de 20 k, especialmente en fluídos corporales. Los anticuerpos descritos se pueden emplear en la detección y cuantificación de hgh de 20k, especialmente en el suero.Peer reviewe

Modified CUPRAC method with electrochemical detection for the determination of antioxidant capacity of gallic acid

A modified cupric reducing antioxidant capacity (CUPRAC) method based on electrochemical monitoring of the Cu(II) ion reduction signal has been developed for the determination of the antioxidant capacity (AOC) of gallic acid. Different electrochemical techniques have been used, namely cyclic voltammetry, differential pulse voltammetry, and square-wave voltammetry (SWV). The SWV method is used in direct scanning and reverse scanning modes. The technique used is SWV in the reverse scanning mode due to the reproducibility of the signal, the sensibility of the technique, and the absence of interferences. The AOC of a given molecule can be measured from the decrease in the reduction signal after the addition of an antioxidant. The relative AOC of gallic versus ascorbic acid obtained by these measurements is 1.9. This value agrees with those obtained from the spectrophotometric methods CUPRAC (1.9) and activity versus 2,2-diphenyl-1-picrylhydrazyl radical (1.8), thus indicating that the method proposed here is valid. Moreover, the electrochemical measurements are not affected by excess or defect due to the turbidity or the colour of samples or the product of the reaction (or both) and are carried out under near-physiological conditions (medium–high ionic strength and absence of non-aqueous solvent)

Modified CUPRAC method with electrochemical detection for the determination of antioxidant capacity of gallic acid

A modified cupric reducing antioxidant capacity (CUPRAC) method based on electrochemical monitoring of the Cu(II) ion reduction signal has been developed for the determination of the antioxidant capacity (AOC) of gallic acid. Different electrochemical techniques have been used, namely cyclic voltammetry, differential pulse voltammetry, and square-wave voltammetry (SWV). The SWV method is used in direct scanning and reverse scanning modes. The technique used is SWV in the reverse scanning mode due to the reproducibility of the signal, the sensibility of the technique, and the absence of interferences. The AOC of a given molecule can be measured from the decrease in the reduction signal after the addition of an antioxidant. The relative AOC of gallic versus ascorbic acid obtained by these measurements is 1.9. This value agrees with those obtained from the spectrophotometric methods CUPRAC (1.9) and activity versus 2,2-diphenyl-1-picrylhydrazyl radical (1.8), thus indicating that the method proposed here is valid. Moreover, the electrochemical measurements are not affected by excess or defect due to the turbidity or the colour of samples or the product of the reaction (or both) and are carried out under near-physiological conditions (medium–high ionic strength and absence of non-aqueous solvent)

Vaccination programs, parity, and calving season as factors affecting the risk of fetal losses and mummified fetuses in Holstein cows

Aim of the study: To investigate vaccination programs, parity, and calving season as factors affecting the risk of abortion and mummified fetuses in Holstein cows.Area of study: Hot zone of Northeast Mexico.Material and methods: Multiple logistic regression models were used to examine the relationship between peripartum disorders, parity, previous occurrence of abortion, season of calving, vaccination program, incidence of abortion, and mummified fetuses in Holstein cows.Main results: For 7014 pregnancies (2886 cows), the percentage of cows aborting and having mummified fetuses was 17.7% and 1.1%, respectively. As the number of brucellosis vaccinations increased, the incidence of abortion increased (10.4% for a single vaccination and 38.0% for 6 accumulated vaccinations). Abortion for cows having 1-2 previous abortions (56%) and >2 abortions (77%) was fivefold and sevenfold greater (p<0.01), respectively, than that for cows without previous abortion. Other important risk factors for abortion were number of calvings (19.8% for nulliparous and primiparous vs. 13.8% for >3 parturitions; OR=1.7, p<0.01), leptospirosis vaccine application <55 days postpartum (dpp; OR=1.3, p<0.05), viral vaccine application >37 dpp (OR=1.3, p<0.01), brucellosis vaccine application >20 dpp (OR=1.6, p<0.01), and no application of clostridial vaccine (OR=3.7, p<0.01). Significant risk factors for mummified fetuses were application of ≥3 brucellosis vaccinations (OR=3.3, p<0.01), no application of 10-way clostridial vaccine (OR=2.3, p<0.01), >2 previous abortions (OR=18.4, p<0.01), and calving in autumn (OR=0.4, compared to winter, p<0.05).Research highlights: Risk of abortion and mummified fetuses in Holstein cows has been found to be related to vaccination programs

Determination of antioxidant activity of spices and their active principles by Differential Pulse Voltammetry

The anodic oxidation of mercury in the presence of hydrogen peroxide in differential pulse voltammetry (DPV) was used to determine the antioxidant (AO) character of radical scavengers. Hydroperoxide radical is formed at the potentials of the oxidation peak on mercury electrodes, such radical reacting with the antioxidants in different extension. The parameter C10 (antioxidant concentration at which the peak area decreases by 10%) is used to measure the scavenging activity of the individual antioxidants. To establish the scavenging activity of antioxidant mixtures as a whole, the parameter, μ10 as the reverse of V10, V10 being the volume necessary to decrease the peak area in DPV by 10%, was selected. Higher μ10 values correspond to higher scavenging activity. The studies have been extended to aqueous extracts of some species. The results may be useful in explaining the effect of spices in vitro and in vivo studies

Functional Inactivation of CXC Chemokine Receptor 4–mediated Responses through SOCS3 Up-regulation

Hematopoietic cell growth, differentiation, and chemotactic responses require coordinated action between cytokines and chemokines. Cytokines promote receptor oligomerization, followed by Janus kinase (JAK) kinase activation, signal transducers and transactivators of transcription (STAT) nuclear translocation, and transcription of cytokine-responsive genes. These include genes that encode a family of negative regulators of cytokine signaling, the suppressors of cytokine signaling (SOCS) proteins. After binding their specific receptors, chemokines trigger receptor dimerization and activate the JAK/STAT pathway. We show that SOCS3 overexpression or up-regulation, stimulated by a cytokine such as growth hormone, impairs the response to CXCL12, measured by Ca2+ flux and chemotaxis in vitro and in vivo. This effect is mediated by SOCS3 binding to the CXC chemokine receptor 4 receptor, blocking JAK/STAT and Gαi pathways, without interfering with cell surface chemokine receptor expression. The data provide clear evidence for signaling cross-talk between cytokine and chemokine responses in building a functional immune system

Growth Hormone Reprograms Macrophages toward an Anti-Inflammatory and Reparative Profile in an MAFB-Dependent Manner

Growth hormone (GH), a pleiotropic hormone secreted by the pituitary gland, regulates immune and inflammatory responses. In this study, we show that GH regulates the phenotypic and functional plasticity of macrophages both in vitro and in vivo. Specifically, GH treatment of GM-CSF–primed monocyte–derived macrophages promotes a significant enrichment of anti-inflammatory genes and dampens the proinflammatory cytokine profile through PI3K-mediated downregulation of activin A and upregulation of MAFB, a critical transcription factor for anti-inflammatory polarization of human macrophages. These in vitro data correlate with improved remission of inflammation and mucosal repair during recovery in the acute dextran sodium sulfate–induced colitis model in GH-overexpressing mice. In this model, in addition to the GH-mediated effects on other immune cells, we observed that macrophages from inflamed gut acquire an anti-inflammatory/reparative profile. Overall, these data indicate that GH reprograms inflammatory macrophages to an anti-inflammatory phenotype and improves resolution during pathologic inflammatory responses.This work was supported in part by grants from the Spanish Ministry of Science, Innovation and Universities (SAF2017-82940-R Agencia Estatal de Investigación/Fondo Europeo de Desarrollo Regional (AEI/FEDER), Unión Europea [UE] [to M.M.], SAF2017-83785-R AEI/FEDER, UE [to Á.L.C.] and FJCI-2016-29990 AEI/FEDER, UE [to B.S.P.]), from the Redes Temáticas de Investigación Cooperativa en Salud Program of Instituto de Salud Carlos III (RD12/0012/0006 and RD12/0012/0007, Red de Investigación en Inflamación y Enfermedades Reumáticas), and the Regional Government of Madrid (B2017/BMD-3804 [to C.M.-A.])

Activation of Th lymphocytes alters pattern expression and cellular location of VIP receptors in healthy donors and early arthritis patients

Vasoactive Intestinal Peptide (VIP) is an important immunomodulator of CD4+ cells in normal and pathological conditions, which exerts its anti-infammatory and immunomodulatory actions through VPAC receptors, VPAC1 and VPAC2. Only a decrease in the expression of VPAC1 mRNA on Th cells upon activation has been reported. Thus, the deepening in the knowledge of the behavior of these receptors may contribute to the design of new therapies based on their activation and/or blockade. In this study, we describe the expression pattern, cellular location and functional role of VIP receptors during the activation of human Th cells in healthy conditions and in early arthritis (EA). The protein expression pattern of VPAC1 did not change with the activation of Th lymphocytes, whereas VPAC2 was up-regulated. In resting cells, VPAC1 was located on the plasma membrane and nucleus, whereas it only appeared in the nucleus in activated cells. VPAC2 was always found in plasma membrane location. VIP receptors signaled through a PKA-dependent pathway in both conditions, and also by a PKAindependent pathway in activated cells. Both receptors exhibit a potent immunomodulatory capacity by controlling the pathogenic profle and the activation markers of Th cells. These results highlight a novel translational view in infammatory/autoimmune diseases