Connexin-43 prevents hematopoietic stem cell senescence through transfer of reactive oxygen species to bone marrow stromal cells

Hematopoietic stem cell (HSC) aging has become a concern in chemotherapy of older patients. Humoral and paracrine signals from the bone marrow (BM) hematopoietic microenvironment (HM) control HSC activity during regenerative hematopoiesis. Connexin-43 (Cx43), a connexin constituent of gap junctions (GJs) is expressed in HSCs, down-regulated during differentiation, and postulated to be a self-renewal gene. Our studies, however, reveal that hematopoietic-specific Cx43 deficiency does not result in significant long-term competitive repopulation deficiency. Instead, hematopoietic Cx43 (H-Cx43) deficiency delays hematopoietic recovery after myeloablation with 5-fluorouracil (5-FU). 5-FU-treated H-Cx43-deficient HSC and progenitors (HSC/P) cells display decreased survival and fail to enter the cell cycle to proliferate. Cell cycle quiescence is associated with down-regulation of cyclin D1, up-regulation of the cyclin-dependent kinase inhibitors, p21cip1. and p16INK4a, and Forkhead transcriptional factor 1 (Foxo1), and activation of p38 mitogen-activated protein kinase (MAPK), indicating that H-Cx43-deficient HSCs are prone to senescence. The mechanism of increased senescence in H-Cx43-deficient HSC/P cells depends on their inability to transfer reactive oxygen species (ROS) to the HM, leading to accumulation of ROS within HSCs. In vivo antioxidant administration prevents the defective hematopoietic regeneration, as well as exogenous expression of Cx43 in HSC/P cells. Furthermore, ROS transfer from HSC/P cells to BM stromal cells is also rescued by reexpression of Cx43 in HSC/P. Finally, the deficiency of Cx43 in the HM phenocopies the hematopoietic defect in vivo. These results indicate that Cx43 exerts a protective role and regulates the HSC/P ROS content through ROS transfer to the HM, resulting in HSC protection during stress hematopoietic regeneration.

Connexin-43 in the osteogenic BM niche regulates its cellular composition and the bidirectional traffic of hematopoietic stem cells and progenitors

Connexin-43 (Cx43), a gap junction protein involved in control of cell proliferation, differentiation and migration, has been suggested to have a role in hematopoiesis. Cx43 is highly expressed in osteoblasts and osteogenic progenitors (OB/P). To elucidate the biologic function of Cx43 in the hematopoietic microenvironment (HM) and its influence in hematopoietic stem cell (HSC) activity, we studied the hematopoietic function in an in vivo model of constitutive deficiency of Cx43 in OB/P. The deficiency of Cx43 in OB/P cells does not impair the steady state hematopoiesis, but disrupts the directional trafficking of HSC/progenitors (Ps) between the bone marrow (BM) and peripheral blood (PB). OB/P Cx43 is a crucial positive regulator of transstromal migration and homing of both HSCs and progenitors in an irradiated microenvironment. However, OB/P Cx43 deficiency in nonmyeloablated animals does not result in a homing defect but induces increased endosteal lodging and decreased mobilization of HSC/Ps associated with proliferation and expansion of Cxcl12-secreting mesenchymal/osteolineage cells in the BM HM in vivo. Cx43 controls the cellular content of the BM osteogenic microenvironment and is required for homing of HSC/Ps in myeloablated animals

Estudio del comportamiento del hormigón de alta resistencia reforzado con fibras de acero frente al impacto de proyectiles

En la presente investigación se buscó estudiar el efecto de la adición de fibras metálicas como refuerzo en hormigones de alta resistencia, y en especial su comportamiento frente al impacto de proyectiles. Se efectuó el estudio sobre un hormigón de alta resistencia (HAR), analizando los aspectos mecánicos, durabilidad y trabajabilidad para su colocación en obra. Las pruebas de laboratorio se llevaron a cabo en el Laboratorio de Materiales de Construcción de la Escuela Técnica Superior de Caminos Canales y Puertos de la UPM y los ensayos balísticos en la galería de tiro cubierta del Polígono de Experiencia de Carabanchel, adscrito a la Dirección General de Infraestructura del Ministerio de la Defensa. La caracterización del HAR empleado en el estudio se centró en los aspectos de resistencias mecánicas a compresión, tracción, flexotracción, tenacidad a flexotracción, punzonamiento, retracción, fluencia, temperatura interna y resistencia al impacto de proyectiles, siempre buscando de manera primordial analizar el efecto de la adición de fibras en el hormigón de alta resistencia. El programa de ensayos balísticos comprendió la fabricación de 47 placas de hormigón de diferentes espesores, desde 5 a 40 cm., 26 de dichas placas eran de HAR con una adición de fibras metálicas de 80 kg/m3, 11 de ellas eran de HAR sin fibras y 10 de un hormigón de resistencia convencional con y sin fibras; sobre dichas placas se efectuaron diversos impactos con proyectiles de los cuatro calibres siguientes: 7.62 AP, 12.70 M8, 20 mm APDS y 25 mm APDS. Las pruebas mostraron que el HAR presenta una mayor resistencia a los impactos de proyectiles, aunque sin la adición de fibras su fragilidad es un serio inconveniente para su utilización como barrera protectora, la adición de fibras reduce considerablemente la fragmentación en la cara posterior “scabbing” y en menor medida en la cara anterior “spalling”. También se incrementa la capacidad del hormigón a la resistencia de múltiples impactos. Se efectuó un estudio de las diferentes formulas y modelos, en especial el modelo desarrollado por Moreno [60], que se vienen utilizando para el diseño de barreras protectoras de hormigón contra impacto de proyectiles, analizando su viabilidad en el caso del hormigón de alta resistencia, hormigón para el cual no fueron desarrolladas y para el que no existen bases de cálculo específicas. In this research we have tried to study the effect of adding metallic fibres as a means of reinforcing high strength concrete, and especially its behaviour when impacted upon by projectiles. The study was carried out using high strength concrete (HSC), analysing its mechanical facets, durability and malleability when used in construction. The laboratory tests took place in the Laboratorio de Materiales de Construcción of the Escuela Técnica Superior de Caminos Canales y Puertos of the Universidad Politécnica de Madrid, and the ballistic tests were carried out in the covered shooting gallery of the Polígono de Experiencias in Carabanchel (Madrid), belongs to the Departamento de Infraestructura of the Ministerio de Defensa. The aspects of the HSC studied are its mechanical strength to compression, traction, flexotraction, resilience to flexo-traction, shear strength, creep, shrinkage, internal temperature and strength to the impact of projectiles, always looking to analyse the effect of adding fibres to HSC. The ballistic testing process required the construction of 47 concrete plates of different thicknesses, from 5 to 40 cm, 26 made which HSC containing of 80 kg/m3 metallic fibres of, 11 made of HSC without fibres, and 10 made with concrete of normal strength with and without fibres. These plates were subjected to a variety of impacts by four projectile, 7.62 AP, 12.70 M8, 20 mm APDS and 25 mm APDS. The results showed that HSC has a greater resistance to the impact of projectiles, although without the addition of fibres, its fragility makes it much less suitable for use as a protective barrier. The addition of fibres reduces considerably frontal fragmentation, known as “scabbing”, and to a lesser extent causes fragmentation of the reverse side, known as “spalling”. In addition, the concrete’s capacity to resist multiple impacts is improved by its letter ductility. A study was carried out on the various formulae and models used to design protective concrete barriers impacted on by projectiles, analysing their viability in the case of HSC for which they were not developed and for which no specific calculations exist.