Hyperglycemia and hyperlipidemia can induce morphophysiological changes in rat cardiac cell line.

H9c2 cardiac cells were incubated under the control condition and at different hyperglycemic and hyperlipidemic media, and the following parameters were determined and quantified: a) cell death, b) type of cell death, and c) changes in cell length, width and height. Of all the proven media, the one that showed the greatest differences compared to the control was the medium glucose (G) 33 mM + 500 µM palmitic acid. This condition was called the hyperglycemic and hyperlipidemic condition (HHC). Incubation of H9c2 cells in HHC promoted 5.2 times greater total cell death when compared to the control. Of the total death ofthe HHC cells, 38.6% was late apoptotic and 8.3% early apoptotic. HHC also changes cell morphology. The reordering of the actin cytoskeleton and cell stiffness was also studied in control and HHC cells. The actin cytoskeleton was quantified and the number and distance of actin bundles were not the same in the control as under HHC. Young's modulus images show a map of cell stiffness. Cells incubated in HHC with the reordered actin cytoskeleton were stiffer than those incubated in control. The region of greatest stiffness was the peripheral zone of HHC cells (where the number of actin bundles was higher and the distance between them smaller). Our results suggest a correlation between the reordering of the actin cytoskeleton and cell stiffness. Thus, our study showed that HHC can promote morphophysiological changes in rat cardiac cells confirming that gluco-and lipotoxicity may play a central role in the development of diabetic cardiomyopathy.

High CO2 Levels Impair Lung Wound Healing.

Delayed lung repair leads to alveolopleural fistulae, which are a major cause of morbidity after lung resections. We have reported that intrapleural hypercapnia is associated with delayed lung repair after lung resection. Here, we provide new evidence that hypercapnia delays wound closure of both large airway and alveolar epithelial cell monolayers because of inhibition of epithelial cell migration. Cell migration and airway epithelial wound closure were dependent on Rac1-GTPase activation, which was suppressed by hypercapnia directly through the upregulation of AMP kinase and indirectly through inhibition of injury-induced NF-κB-mediated CXCL12 (pleural CXC motif chemokine 12) release, respectively. Both these pathways were independently suppressed, because dominant negative AMP kinase rescued the effects of hypercapnia on Rac1-GTPase in uninjured resting cells, whereas proteasomal inhibition reversed the NF-κB-mediated CXCL12 release during injury. Constitutive overexpression of Rac1-GTPase rescued the effects of hypercapnia on both pathways as well as on wound healing. Similarly, exogenous recombinant CXCL12 reversed the effects of hypercapnia through Rac1-GTPase activation by its receptor, CXCR4. Moreover, CXCL12 transgenic murine recipients of orthotopic tracheal transplantation were protected from hypercapnia-induced inhibition of tracheal epithelial cell migration and wound repair. In patients undergoing lobectomy, we found inverse correlation between intrapleural carbon dioxide and pleural CXCL12 levels as well as between CXCL12 levels and alveolopleural leak. Accordingly, we provide first evidence that high carbon dioxide levels impair lung repair by inhibiting epithelial cell migration through two distinct pathways, which can be restored by recombinant CXCL12.

17,20 beta-P and cortisol are the main in vitro metabolites of 17-hydroxy-progesterone produced by spermiating testes of Micropogonias furnieri (Desmarest, 1823) (Perciformes Sciaenidae)

The aim was to investigate the major C21 steroids produced by spermiating white croaker Micropogonias furnieri (Sciaenidae) in order to establish the potential mediator of gamete maturation in males of this species. The testes steroid production at the spawning season was identified incubating the 3H-17-hydroxy-4-pregnene-3,20-dione precursor through thin layer chromatography, high pressure liquid chromatography, enzymatic oxydation, acetylation and immunochemistry analyses. 17,20β-Dihydroxy-4-pregnen-3-one (17,20β-P) and 11β,17,21-Trihydroxy-4-pregnene-3,20-dione (cortisol) were the main metabolites produced. Contrary to what we expected, 17,20β,21-Trihydroxy-4-pregnen-3-one was not detected. Circulating levels of 17,20β-P were undetectable in immature testes and in those at the first spermatogenesis stages, while a clear increase was observed during the whole spermatogenesis and spermiation phases (from undetectable to 1047 pg mL-1). In vitro studies together with plasma detection suggest that 17,20β-P is a good steroid candidate involved in M. furnieri testes maturation. The role of cortisol during late phases of testes development needs further studies.

El objetivo fue investigar cuales eran los esteroides C21 más importantes producidos por los testículos en espermiación de la corvina blanca Micropogonias furnieri (Sciaenidae) para poder identificar los potenciales mediadores de la maduración gamética de los machos de esta especie. Los esteroides producidos por los testículos en espermiación fueron identificados después de su incubación con el precursor 3H-17-hidroxi-4-pregnene-3,20-diona a través de cromatografía de capa fina y cromatografía líquida de alta presión y posteriormente por oxidación enzimática, acetilación e inmunoquímica. Los principales metabolitos producidos por los testículos en espermiación fueron la 17,20β-Dihidroxi-4-pregnen-3-ona (17,20β-P) y la 11β,17,21-Trihidroxi-4-pregnene-3,20-diona (cortisol). Contrariamente a lo esperado, no se encontró el derivado tri-hidroxilado de la progesterona llamado 17,20β,21-Trihidroxi-4-pregnen-3-ona. Los niveles circulantes de 17,20β-P fueron no detectable en los testículos inmaduros y en aquellos en inicios de espermatogénesis, mientras que un aumento claro en las concentraciones circulantes fue detectada en corvinas en plena espermatogénesis y en espermiación (desde no detectable a 1047 pg mL-1). Los resultados obtenidos in vitrojunto a los cambios a nivel plasmático sugieren que la 17,20β-P es un buen candidato a proponer como esteroide involucrado en la regulación del proceso de maduración testicular de la corvina. La función del cortisol a nivel testicular debería ser mejor estudiada en las etapas finales del desarrollo testicular de esta especie.