Expresión del factor de crecimiento del endotelio vascular (VEGF) en el riñón de ratones adultos

Bajo condiciones normales menos del 1% de las células tubulares del riñón proliferan aunque sin embargo, en respuesta a una injuria, células normalmente quiescentes entran en el ciclo celular. En el modelo del riñón remanente en roedores, el número de nefronas es repentinamente reducido por ablación quirúrgica, lo que dispara eventos moleculares y celulares que promueven el crecimiento compensatorio. Históricamente han surgido controversias acerca de si dicho crecimiento renal resulta de hipertrofia o hiperplasia. Por otro lado, además de los cambios en las células epiteliales e intersticiales después de la reducción de la masa renal, la reparación capilar es un evento crucial en la recuperación del daño renal y el factor de crecimiento del endotelio vascular (VEGF) juega un rol importante en la proliferación endotelial. En el riñón normal, el VEGF se expresa en los podocitos glomerulares y en las células tubulares, especialmente en la médula externa y rayos medulares, pero también ha sido demostrado que juega un rol mayor en la respuesta compensatoria renal después de la uninefrectomía. Si bien este factor es esencial para la normal nefrogénesis y la glomerulogénesis, también ha sido implicado en la patogénesis de la disfunción renal temprana y en la hipertrofia glomerular en la diabetes experimental. Además, los cambios en los capilares peritubulares después de la reducción renal son modulados por distintas causas como, la especie y la edad de los animales bajo estudio, la extensión y el origen de la reducción néfrica, el tiempo posterior a la injuria y el grado de fibrosis y/o de proliferación tubular. Existen indicaciones de que las hormonas sexuales tienen distintos efectos según las regiones del riñón y posiblemente también durante el crecimiento compensatorio después de la uninefrectomía. Conjuntamente, durante algunos procesos regenerativos en ratas y ratones, la expresión de ARNm VEGF está temporal y espacialmente relacionada a la proliferación.Facultad de Ciencias Médica

Expression of vascular endothelial growth factor in normal and tumoral liver mouse cells

In previous studies have been reported a controversy between the reactivity of mono and polyclonal antibodies in different cells populations of the mouse. In this study we focus on the immunoreactivity of the monoclonal antibody VEGF (C- 1) in order to analyze its expression in two cell populations of regenerating mouse liver (hepatocytes and endothelial cells) after partial hepatectomy, and two transplanted hepatocarcinomas (ES2 and SS1K). C3H/S male mice were divided into two groups and kept under standard conditions for circadian periodicity analysis, one group were transplanted with the tumors into the subcutaneous tissue of the animal flank and the other group were subjected to a partial hepatectomy (70 %). The animals were killed by decapitation and exsanguination. Samples of regeneration liver, ES2 and SS1K tumors were processed for histology and immunostained with VEGF (C-1) mouse monoclonal IgG2a antibody. The results showed strongly immunopositivity reaction for VEGF either in hepatocytes or endothelial cells of mouse regenerating liver. We also demonstrated positive immunoreaction in both malignant tumors ES2 and SS1K endothelial and parenchymal cells. We conclude that the use of VEGF (C-1) mouse monoclonal IgG2a antibody can be applied successfully in experimental design leading to study processes involved in mice’s angiogenesi

Cochleates derived from Vibrio cholerae O1 proteoliposomes : The impact of structure transformation on mucosal immunisation

Cochleates are phospholipid-calcium precipitates derived from the interaction of anionic lipid vesicles with divalent cations. Proteoliposomes from bacteria may also be used as a source of negatively charged components, to induce calcium-cochleate formation. In this study, proteoliposomes from V. cholerae O1 (PLc) (sized 160.7±1.6 nm) were transformed into larger (16.3±4.6 µm) cochleate-like structures (named Adjuvant Finlay Cochleate 2, AFCo2) and evaluated by electron microscopy (EM). Measurements from transmission EM (TEM) showed the structures had a similar size to that previously reported using light microscopy, while observations from scanning electron microscopy (SEM) indicated that the structures were multilayered and of cochleate-like formation. The edges of the AFCo2 structures appeared to have spaces that allowed penetration of negative stain or Ovalbumin labeled with Texas Red (OVA-TR) observed by epi-fluorescence microscopy. In addition, freeze fracture electron microscopy confirmed that the AFCo2 structures consisted of multiple overlapping layers, which corresponds to previous descriptions of cochleates. TEM also showed that small vesicles co-existed with the larger cochleate structures, and in vitro treatment with a calcium chelator caused the AFCo2 to unfold and reassemble into small proteoliposome-like structures. Using OVA as a model antigen, we demonstrated the potential loading capacity of a heterologous antigen and in vivo studies showed that with simple admixing and administration via intragastric and intranasal routes AFCo2 provided enhanced adjuvant properties compared with PLc

Estudio comparativo de la síntesis de ADN en hígado y riñón de ratones lactantes y adultos

In previous papers we have observed that the proliferative activity of numerous cell populations of suckling and young mice was different than the observed in adults. In the present work we analized the DNA synthesis of suckling and adult mice, by immunohistochemistry (Bromodioxiuridine) in both hepatocytes and renocytes, at two different time points. The object of the present experiment was to establish the correlation in cellular proliferation between both groups. We used C3HS suckling male mice of 21 days old and adult mice of 90 days old. The results showed that DNA synthesis of renocytes and hepatocytes in suckling mice were significantly higher than the adults, and we didn't find statistically differences between the values of both analysed time points. We can conclude that in suckling mice we could observe maximum and minimum values but we coudn't find a circadian rhythm like those established in adults.En trabajos previos hemos observado que la actividad proliferativa de numerosas poblaciones celulares del ratón lactante y joven difiere de la encontrada en el adulto. En el presente trabajo experimental, se estudia la síntesis de ADN del ratón lactante y adulto, mediante el método inmunohistoquímico de la Brdu (Bromodeoxiuridina), en las poblaciones celulares de hepatocitos y renocitos, a las OO:O0 h y 16:00 h, con el objeto de establecer la correspondencia en la proliferación celular entre ambos grupos etarios. Para ello se utilizaron machos lactantes de 21 días de edad y adultos de 90 días, de la cepa C3H/S. Los resultados muestran que los valores de síntesisde ADN de los renocitos y hepatodtos en los ratones lactantes son significativamente mayores (p<O.001) a los observados en los adultos, no encontrándose diferencias estadísticamente significativas entre los valores de ambos puntos horarios en los órganos estudiados. Podemos concluir que estos hallazgos demuestran que, si bien en los ratones lactantes aparecen valores máximos y mínimos en la actividad proliferativa de distintas poblaciones celulares, estas aún no presentan la ritmicidad establecida para los adultos

Expresión del factor de crecimiento del endotelio vascular (VEGF) durante el crecimiento compensatorio del riñón

El objetivo de este trabajo es estudiar la expresión del VEGF en los túbulos de la corteza y de la médula externa del riñón de ratones hembra después de la nefrectomía izquierda.Facultad de Ciencias Médica

Expression of vascular endothelial growth factor in normal and tumoral liver mouse cells

In previous studies have been reported a controversy between the reactivity of mono and polyclonal antibodies in different cells populations of the mouse. In this study we focus on the immunoreactivity of the monoclonal antibody VEGF (C-1) in order to analyze its expression in two cell populations of regenerating mouse liver (hepatocytes and endothelial cells) after partial hepatectomy, and two transplanted hepatocarcinomas (ES2 and SS1K). C3H/S male mice were divided into two groups and kept under standard conditions for circadian periodicity analysis, one group were transplanted with the tumors into the subcutaneous tissue of the animal flank and the other group were subjected to a partial hepatectomy (70 %). The animals were killed by decapitation and exsanguination. Samples of regeneration liver, ES2 and SS1K tumors were processed for histology and immunostained with VEGF (C-1) mouse monoclonal IgG2a antibody. The results showed strongly immunopositivity reaction for VEGF either in hepatocytes or endothelial cells of mouse regenerating liver. We also demonstrated positive immunoreaction in both malignant tumors ES2 and SS1K endothelial and parenchymal cells. We conclude that the use of VEGF (C-1) mouse monoclonal IgG2a antibody can be applied successfully in experimental design leading to study processes involved in mice’s angiogenesisFacultad de Ciencias Médica

Women becoming social justice leaders with an inclusive view in Costa Rica, Mexico and Spain

This study looks at three female school directors in Costa Rica, Mexico, and Spain who worked under challenging conditions to establish social justice. We were particularly interest in how they learned to become social justice leaders. Qualitative interviews were used to hear directly from the school directors about their experiences. Transcripts were analyzed for common themes. The commitment of these directors to social justice came from early family experiences that gave them strength and core values. They met adversity in young adulthood which reinforced their commitment to inclusive leadership

Evaluation of angiogenesis with the expression of VEGF and CD34 in human non-small cell lung cancer

Angiogenesis is an essential process in the progression of malignant tumors and the most potent angiogenic factor is the vascular endothelial growth factor (VEGF). On the other hand, the CD34 is an endothelial antigen that has been used to highlight the microvasculature vessel density (MVD) as a direct marker of the degree of neoangiogenesis. In the present study we report the VEGF expression and its relationship with MVD, measured by CD34, in two lineages of non-small cell lung cancer (NSCL): low differentiated adenocarcinomas and epidermoid carcinomas, in order to consider the possibility of using the correlation between both antibodies as a prognostic factor. Tumor sections were stained by immunohistochemistry for CD34 and VEGF. The results showed that the mean value of VEGF for adenocarcinoma was significantly higher than the one for epidermoid carcinoma (p < 0.001). However, the mean of MVD did not show significant differences between both types of tumors. The conventional factors taken into consideration (age over 60, sex, and presence of lymph nodes) was not significantly related to the angiogenic factors examined. In conclusion, we could affirm that CD34 is a better prognostic marker of neoangiogenesis in NSCLC, because both types of tumors have the same clinical prognosis, and so we expected the same behaviour from both markers.Facultad de Ciencias Médica

Evaluation of angiogenesis with the expression of VEGF and CD34 in human non-small cell lung cancer

Angiogenesis is an essential process in the progression of malignant tumors and the most potent angiogenic factor is the vascular endothelial growth factor (VEGF). On the other hand, the CD34 is an endothelial antigen that has been used to highlight the microvasculature vessel density (MVD) as a direct marker of the degree of neoangiogenesis. In the present study we report the VEGF expression and its relationship with MVD, measured by CD34, in two lineages of non-small cell lung cancer (NSCL): low differentiated adenocarcinomas and epidermoid carcinomas, in order to consider the possibility of using the correlation between both antibodies as a prognostic factor. Tumor sections were stained by immunohistochemistry for CD34 and VEGF. The results showed that the mean value of VEGF for adenocarcinoma was significantly higher than the one for epidermoid carcinoma (p < 0.001). However, the mean of MVD did not show significant differences between both types of tumors. The conventional factors taken into consideration (age over 60, sex, and presence of lymph nodes) was not significantly related to the angiogenic factors examined. In conclusion, we could affirm that CD34 is a better prognostic marker of neoangiogenesis in NSCLC, because both types of tumors have the same clinical prognosis, and so we expected the same behaviour from both markers.Facultad de Ciencias Médica

Barley-ß-glucans reduce systemic inflammation, renal injury and aortic calcification through ADAM17 and neutral-sphingomyelinase2 inhibition

In chronic kidney disease (CKD), hyperphosphatemia-induced inflammation aggravates vascular calcification (VC) by increasing vascular smooth muscle cell (VSMC) osteogenic differentiation, ADAM17-induced renal and vascular injury, and TNFα-induction of neutral-sphingomyelinase2 (nSMase2) to release pro-calcifying exosomes. This study examined anti-inflammatory β-glucans efficacy at attenuating systemic inflammation in health, and renal and vascular injury favoring VC in hyperphosphatemic CKD. In healthy adults, dietary barley β-glucans (Bβglucans) reduced leukocyte superoxide production, inflammatory ADAM17, TNFα, nSMase2, and pro-aging/pro-inflammatory STING (Stimulator of interferon genes) gene expression without decreasing circulating inflammatory cytokines, except for γ-interferon. In hyperphosphatemic rat CKD, dietary Bβglucans reduced renal and aortic ADAM17-driven inflammation attenuating CKD-progression (higher GFR and lower serum creatinine, proteinuria, kidney inflammatory infiltration and nSMase2), and TNFα-driven increases in aortic nSMase2 and calcium deposition without improving mineral homeostasis. In VSMC, Bβglucans prevented LPS- or uremic serum-induced rapid increases in ADAM17, TNFα and nSMase2, and reduced the 13-fold higher calcium deposition induced by prolonged calcifying conditions by inhibiting osteogenic differentiation and increases in nSMase2 through Dectin1-independent actions involving Bβglucans internalization. Thus, dietary Bβglucans inhibit leukocyte superoxide production and leukocyte, renal and aortic ADAM17- and nSMase2 gene expression attenuating systemic inflammation in health, and renal injury and aortic calcification despite hyperphosphatemia in CKD.A grant to A.S.D. and M.J.M. from IRBLleida and Agrotecnio Research collaborative projects from the Consell Social at Lleida University supported initial work, Instituto de Salud Carlos III and co-funded by European Union (ERDF/FEDER) (FIS PI11/00259, PI14/01452, PI17/02181), Plan de Ciencia, Tecnología e Innovación 2013–2017 y 2018–2022 del Principado de Asturias (GRUPIN14-028, IDI-2018-000152), RedInRen from ISCIII (ISCIII-RETIC REDINREN RD16/0009). Investigator support included: NC-L by GRUPIN14-028 and IDI-2018-000152, LM-A by GRUPIN14-028, SP by FICYT; MVA and PV by Educational Grant 2 A/2015 from ERA-EDTA CKD-MBD Working Group; PV and AC by ERA-EDTA fellowships 2011 and 2012; JR-C by MINECO (“Juan de la Cierva” program, FJCI-2015-23849); A.S.D. by Asociación Investigación de Fisiología Aplicada. A.S.D. and M.J.M. are members of the Campus Iberus (Ebro Valley Campus of International Excellence)