Hepatocyte growth factor is elevated in amniotic fluid from obese women and regulates placental glucose and fatty acid metabolism.

INTRODUCTION: To evaluate the impact of the pro-inflammatory cytokine hepatocyte growth factor (HGF) on the regulation of glucose and lipid placental metabolism. METHODS: HGF levels were quantified in amniotic fluid and placenta from control and obese women. 2-deoxy-glucose (2-DOG) uptake, glycolysis, fatty acid oxidation (FAO), fatty acid esterification, de novo fatty acid synthesis, triglyceride levels and carnitine palmitoyltransferase activities (CPT) were measured in placental explants upon addition of pathophysiological HGF levels. RESULTS: In obese women, total- and -activated-HGF levels in amniotic fluid were elevated ∼24%, and placental HGF levels were ∼3-fold higher than in control women. At a similar dose to that present in amniotic fluid of obese women, HGF (30 ng/mL) increased Glut-1 levels and 2-DOG uptake by ∼25-30% in placental explants. HGF-mediated effect on 2-DOG uptake was dependent on the activation of phosphatidylinositol 3-kinase signaling pathway. In addition, HGF decreased ∼20% FAO, whereas esterification and de novo fatty acid synthesis increased ∼15% and ∼25% respectively, leading to 2-fold triglyceride accumulation in placental explants. In parallel, HGF reduced CPT-I activity ∼70%. DISCUSSION: HGF is a cytokine elevated in amniotic fluid and placental tissue of obese women, which through its ability to stimulate 2-DOG uptake and metabolism impairs FAO and enhances esterification and de novo fatty acid synthesis, leading to accumulation of placental triglycerides.

Búsqueda de nuevos autoantígenos en el síndrome de Sjögren / The search for new autoantigens in Sjögren’s syndrome

Objetivo: Determinar la existencia de nuevos autoantígenos en el síndrome de Sjögren (SS) así como estudiar la prevalencia de éstos en pacientes y población sana. Material y métodos: Se procedió a realizar un muestreo con el suero de una enferma afectada de SS mediante la utilización de una genoteca de cerebro humano (técnica SEREX). Se determinó que este suero expresaba autoantígenos ya conocidos y proteínas no descritas previamente, y también se confirmó la presencia de una proteína desconocida hasta ahora. De entre ellas, se seleccionó a esta última (hIscA) y a la proteína Tau (hallada en el muestreo) para ser transformadas en sendos plásmidos de expresión para conseguir su síntesis recombinante. Resultados: Mediante técnica de inmunotransferencia se testó la existencia de anticuerpos anti-Tau y anti-hIscA en 19 pacientes y 20 sujetos sanos. No se encontró diferencia estadísticamente significativa entre pacientes y controles en la expresión de anticuerpos anti-Tau y se halló que los pacientes expresaban, de forma significativa, valores inferiores de anticuerpos anti-hIscA. Conclusión: Se ha identificado a las proteínas Tau y hIscA como nuevos autoantígenos en el SS. Se ha hallado que los pacientes con SS expresan valores inferiores de anticuerpos anti-hIscA en comparación con controles y, aunque no se ha encontrado ninguna diferencia entre sanos y enfermos en relación con la presencia de anticuerpos anti-Tau, ésta es la primera vez en que anticuerpos contra esta proteína se han detectado en el SS(AU)

Objective: To identify new autoantigens related to Sjögren’s syndrome and to determine their prevalence in patients and healthy individuals. Material and methods: Serological sampling was performed in a patient with Sjögren’s syndrome through the use of a human brain expression genotec (SEREX technique) to determine expression of known autoantigens and previously undescribed proteins. The presence of a previously unknown protein was found. Several proteins were obtained and two were selected to be studied (a human protein called Tau and an unknown protein described by our group and named hlscA). Both Tau and hIscA cDNA were transformed into an expression plasmid to obtain their recombinant proteins. Results: Using a Western-blot technique we investigated the presence of anti-Tau and anti-hlscA autoantibodies in the sera of 19 patients with Sjögren’s syndrome and in the sera of 20 controls. No statistically significant differences were found in the expression of anti-Tau antibodies between patients with Sjögren’s syndrome and controls but values of anti-hlscA autoantibodies were significantly lower in patients with Sjögren’s syndrome. Conclusion: We identified Tau and hIscA proteins as new autoantigens in Sjögren’s syndrome. Anti-hlscA antibody values were significantly lower in patients with Sjögren’s syndrome than in healthy controls. Although no statistically significant differences in values of anti-Tau antibodies were found between Sjögren’s syndrome patients and controls, this is the first time antibodies against this protein have been detected in Sjögren’s síndrome(AU)

[The search for new autoantigens in Sjögren's syndrome]. / Búsqueda de nuevos autoantígenos en el síndrome de Sjögren.

OBJECTIVE: To identify new autoantigens related to Sjögren's syndrome and to determine their prevalence in patients and healthy individuals. MATERIAL AND METHODS: Serological sampling was performed in a patient with Sjögren's syndrome through the use of a human brain expression genotec (SEREX technique) to determine expression of known autoantigens and previously undescribed proteins. The presence of a previously unknown protein was found. Several proteins were obtained and two were selected to be studied (a human protein called Tau and an unknown protein described by our group and named hlscA). Both Tau and hIscA cDNA were transformed into an expression plasmid to obtain their recombinant proteins. RESULTS: Using a Western-blot technique we investigated the presence of anti-Tau and anti-hlscA autoantibodies in the sera of 19 patients with Sjögren's syndrome and in the sera of 20 controls. No statistically significant differences were found in the expression of anti-Tau antibodies between patients with Sjögren's syndrome and controls but values of anti-hlscA autoantibodies were significantly lower in patients with Sjögren's syndrome. CONCLUSION: We identified Tau and hIscA proteins as new autoantigens in Sjögren's syndrome. Anti-hlscA antibody values were significantly lower in patients with Sjögren's syndrome than in healthy controls. Although no statistically significant differences in values of anti-Tau antibodies were found between Sjögren's syndrome patients and controls, this is the first time antibodies against this protein have been detected in Sjögren's syndrome.

Expression and cellular localization of Na,K-ATPase isoforms in the rat ventral prostate.

OBJECTIVE: To determine the expression and plasma membrane domain location of isoforms of Na,K-ATPase in the rat ventral prostate. MATERIALS AND METHODS: Ventral prostate glands from adult male rats were dissected, cryosectioned (7 micro m) and attached to poly-l-lysine coated glass slides. The sections were then fixed in methanol and subjected to indirect immunofluorescence and immunoperoxidase procedures using a panel of well-characterized monoclonal and polyclonal antibodies raised against known Na,K-ATPase subunit isoforms. Immunofluorescence micrographs were digitally captured and analysed by image analysis software. RESULTS: There was expression of Na,K-ATPase alpha1, beta1, beta2 and beta3 subunit isoforms in the lateral and basolateral plasma membrane domains of prostatic epithelial cells. The alpha1 isoform was abundant but there was no evidence of alpha2, alpha3 or gamma isoform expression in epithelial cells. The alpha3 isoform was not detected, but there was a relatively low level of alpha2 isoform expression in the smooth muscle and stroma. CONCLUSION: Rat prostate Na,K-ATPase consists of alpha1/beta1, alpha1/beta2 and alpha1/beta3 isoenzymes. These isoform proteins were located in the lateral and basolateral plasma membrane domains of ventral prostatic epithelial cells. The distribution and subcellular localization of Na,K-ATPase is different in rodent and human prostate. Basolateral Na,K-ATPase probably contributes to the establishment of transepithelial ionic gradients that are a prerequisite for the uptake of metabolites by secondary active transport mechanisms and active citrate secretion.

Na+, K+-ATPase isozyme diversity; comparative biochemistry and physiological implications of novel functional interactions.

Na+, K+-ATPase is ubiquitously expressed in the plasma membrane of all animal cells where it serves as the principal regulator of intracellular ion homeostasis. Na+, K+-ATPase is responsible for generating and maintaining transmembrane ionic gradients that are of vital importance for cellular function and subservient activities such as volume regulation, pH maintenance, and generation of action potentials and secondary active transport. The diversity of Na+, K+-ATPase subunit isoforms and their complex spatial and temporal patterns of cellular expression suggest that Na+, K+-ATPase isozymes perform specialized physiological functions. Recent studies have shown that the alpha subunit isoforms possess considerably different kinetic properties and modes of regulation and the beta subunit isoforms modulate the activity, expression and plasma membrane targeting of Na+, K+-ATPase isozymes. This review focuses on recent developments in Na+, K+-ATPase research, and in particular reports of expression of isoforms in various tissues and experiments aimed at elucidating the intrinsic structural features of isoforms important for Na+, K+-ATPase function.