Zinc Supplementation and Ischemia Pre-conditioning in Renal Ischemia/ Reperfusion Injury

@#Backgrounds: Renal ischemia/reperfusion (RIR) is a major cause of kidney dysfunction in clinic. The main objective of this study was to investigate the effect of pre-conditioning ischemia (IPC) and zinc (Zn) supplementation on renal RIR injury. Methods: A total of 63 unilateral nephrectomised male and female Wistar rats were divided into five groups. Group 1 (ShOPR): Rats as sham-operated group were subjected to surgical procedure without RIR. Group 2 (Isch): Rats underwent RIR (left kidney ischemia for 30 min followed by 48 h reperfusion). Group 3 (Zn+Isch): Rats were treated as group 2 but they received Zn sulphate (30 mg/kg) 1 h before induction of RIR. Group 4 (IPC+Isch): Rats were treated as group 2 but they underwent 1 min of ischemia followed by 3 min reperfusion as IPC, which was repeated for three times before induction of RIR. Group 5 (Zn+IPC+Isch): Rats were subjected to receive both Zn sulphate and IPC before induction of RIR. Urine samples were collected in the last 6 h of reperfusion, and finally biochemical and histological measurements were performed. Results: The serum level of creatinine (Cr), normalised kidney weight (KW) and kidney tissue damage score (KTDS) increased by RIR alone significantly (P < 0.05). These parameters were attenuated statistically by Zn supplementation (P < 0.05). However, IPC alone or cotreatment of Zn and IPC did not improve the biochemical and histological markers altered by RIR injury. Conclusion: Zn supplementation had a protective role against RIR while such protective effect was not observed by IPC alone or by co-treatment of Zn and IPC.

Regular transient limb ischemia prevents atherosclerosis progression in hypercholesterolemic rabbits / 中华医学杂志(英文版)

Background:@#Endothelial dysfunction, the initial pathogenic factor in atherosclerosis, can be alleviated via transient limb ischemia. We observed the effects of regular transient limb ischemia (RTLI) on atherosclerosis in hypercholesterolemic rabbits.@*Methods:@#Twenty-eight rabbits were randomized to control, cholesterol, sham, ischemia groups (n=7 each) between October 2010 and March 2011. They were fed a normal diet in the control group and hypercholesterolemic diet in other groups for 12 weeks. Six cycles of RTLI were performed once per day on the ischemia group. Serum samples were prepared to measure the total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) before the experiment (W0), at the end of weeks 4, 8, 12 (W4, W8, W12). The whole aorta was harvested at W12 and stained using Sudan IV to identify the plaque. The plaque area was measured using Image J. Results were analyzed by analysis of variance or rank sum test.@*Results:@#Concentrations of TC in the cholesterol group were higher than those in the control group at W4 (29.60 [23.75, 39.30] vs. 1.00 [0.80, 1.55], Z = –2.745, P = 0.006), W8 (41.78 [28.08, 47.37] vs. 0.35 [0.10, 0.68], Z = –2.739, P = 0.006), W12 (48.32 [40.04, 48.95] vs. 0.61 [0.50, 0.86], Z = –2.739, P = 0.006). Similar results were obtained for HDL-C and LDL-C. Serum concentrations of TC, HDL-C, and LDL-C in the hypercholesterolemic groups had no differences (all P > 0.05). The percentage of plaque area in the cholesterol group was higher than that in the control group (47.22 ± 23.89% vs. 0, Z = –2.986, P = 0.003). Square root of the percentage of plaque area was smaller in the ischemia group than that in the cholesterol (0.44 ± 0.13 vs. 0.67 ± 0.18, P = 0.014) or sham groups (0.44 ± 0.13 vs. 0.61 ± 0.12, P = 0.049).@*Conclusion:@#In hypercholesterolemic rabbits, RTLI might prevent atherosclerosis progression by reducing the percentage of plaque area.

Cardioprotection of Rosuvastatin Pre-conditioning on Myocardial Ischemia / Reperfusion Injury in a Rat Model

Abstract This study aimed to investigate the cardioprotection of rosuvastatin pre-conditioning (R-Pre) in a rat model of myocardial ischemia / reperfusion (I/R). Male SD rats were assigned into three groups: sham group, I/R group and R-Pre group. Rats in I/R group and R-Pre group received ischemia for 30 min and reperfusion for 2 h. In R-Pre group, rats received intragastrical administration with rosuvastatin at 5 mg/kg once daily for 1 week. After 2-h reperfusion, the cardiac function was detected by ultrasonography; the blood was collected for biochemical analysis; the heart was collected for the TUNEL staining and immunohistochemistry for Bcl-2 and Bax. Our results showed rosuvastatin pre-conditioning for 1 week could significantly reduce the infarct ratio and improve the cardiac function after myocardial I/R injury, in which attenuation of oxidative stress and cell apoptosis played an important role. Our study provides evidence on the cardioprotection of rosuvastatin pre-conditioning and highlight the use of rosuvastatin before cardiopulmonary bypass.

Dexmedetomidina genera óxido nítrico mediante un mecanismo independiente de la óxido nítrico sintasa inducible / Dexmedetomidine protects the heart against ischemia-reperfusion injury by an endothelial eNOS/NO dependent mechanism

El infarto del miocardio es una de las principales causas de muerte a nivel mundial y se produce a consecuencia de procesos de isquemia-reperfusión (IR). El daño miocárdico generado por IR puede ser atenuado a través del pre-condicionamiento isquémico (PI) temprano, mediado por la vía RISK o PI tardío, que se asocia a una respuesta genómica en la que se activan proteínas como óxido nítrico sintasa inducible (iNOS). Las vías de señalización que median el PI también pueden ser activadas farmacológicamente. Dexmedetomi-dina (Dex) es un agonista alfa2-adrenérgico, que se ha descrito como un potente agente cardioprotector frente a IR. Recientemente, nuestro grupo describió que Dex requiere el endotelio y la activación de la vía óxido nítrico sintasa endotelial (eNOS)-óxido nítrico (NO) para pre-condicionar el miocardio. Sin embargo, no existen estudios que muestren la posible participación de iNOS en la protección conferida por Dex. La presente adenda tiene por objetivo evaluar si Dex activa iNOS en el corazón y en cardiomiocitos. Para esto, corazones de rata adulta fueron estimulados con Dex 10 nM y se observó que el fármaco aumentó la producción de NO medida por cuantificación de nitritos, mas no estimuló la activación de iNOS medida por Western blot. Además, Dex tampoco indujo el aumento de mRNA de iNOS en cardiomiocitos adultos. Por lo tanto, Dex genera NO independiente a iNOS durante su efecto pre-condicionante agudo. Sin embargo, se requieren más estudios que clarifiquen su papel en una posible protección a largo plazo frente a IR generada por Dex.

Myocardial infarction is one of the leading causes of death worldwide and is generated as a consequence of ischemia-reperfusion (IR). Myocardial damage inflicted by IR can be attenuate by early ische-mic pre-conditioning (IP), which is mediated by the RISK pathway or late IP, which is associated to a genomic response involving the activation of proteins such as inducible nitric oxide synthase (iNOS). The signaling pathways mediating IP can also be pharmacologically activated. Dexmedetomidine (Dex) is an alpha2-adrenergic receptor agonist, which has been described as a strong cardio protective agent against IR. Recently, our group reported that Dex requires the endothelium and the activation of the endothelial nitric oxide synthase (eNOS)-ni-tric oxide (NO) pathway to precondition the myocardium. However, there are no studies showing the involvement of iNOS in the protection elicited by Dex. The aim of this Addendum is to evaluate if Dex activates iNOS in the heart and cardiomyocytes. To test this, adult rat hearts were stimulated with Dex 10 nM and we observed that NO production measured by quantification of nitrites was increased, but Dex did not activate iNOS measured by Western blot. Moreover, Dex did not induce an increase in the mRNA levels of iNOS in adult cardiomyocytes. Therefore, Dex generates NO independent of iNOS during its early pre-conditioning effect. Nevertheless, more studies are required to clarify its role in a possible long term protection against IR generated by Dex.

Dupla filtração modificada utilizando filtro de pressão a montante de filtro ascendente no tratamento de águas eutrofizadas / Modified double filtration using pressured pre-filter before upflow filter in eutrophic reservoirs

A implantação de uma unidade de pré-tratamento a montante de um filtro ascendente (FA) existente objetiva condicionar a água bruta eutrofizada à capacidade de tratamento do FA. Essa solução esbarra na necessidade de se instalar essa unidade de pré-tratamento com cota mais elevada que o filtro a jusante. Nesta pesquisa, a instalação de um filtro de pressão de fluxo descendente possibilitou a eliminação do problema do desnível geométrico e a adequação da água bruta com elevada presença de fitoplâncton às limitações da filtração direta ascendente (FDA). O pré-filtro de pressão com melhor eficiência, o PFP1, apresentou uma remoção de 50% de cor aparente e 63% de turbidez. A eficiência global do sistema PFP/FA foi satisfatória, apresentando turbidez final inferior a 0,5 uT, cor aparente em torno de 5 uH, além de reduzir as perdas da água tratada de 11,7 para 4,5%.

The implementation of a pretreatment unit before existing upward filter, to condition the eutrophic raw waters to its capacity, is an option which runs into a problem of technical nature, the need of a pretreatment unit with enough hydraulic load to properly feed the existing filter. Typically, the available studies on double filtration present upward filters followed by descendant ones. In this study, the installation of a pressure filter downflow adapted the raw water with high algae concentration to the upward direct filtration system limitations, improving the quality of water produced and reducing losses by washing. The pressure filter with better efficiency was the PFP1 filter showed a removal of 50% of apparent color and 63% of turbidity. The overall efficiency of the PFP/FA system was satisfactory, presenting final turbidity of less than 0.5 uT, apparent color around 5 uH and reducing losses from 11.7 to 4.5% of the total treated water.

Screening and bioinformatics analysis of differentially expressed miRNAs induced by hypoxia preconditioning in rat cardiomyocytes / 中国药理学通报

Aim To screen the differentially expressed microRNAs ( miRNAs ) induced by hypoxia precondi-tioning ( HPC ) in adult rat cardiomyocytes, and pre-dict miRNAs-regulated target genes and their func-tions. Methods Cardiomyocytes were isolated from a-dult rat ventricular myocardium and cultured ( in vitro) . The cells were divided into 2 groups: control group ( CON ) and hypoxia preconditioning group ( HPC) . The cardiomyocytes in HPC group were sub-jected to 10 min hypoxia followed by 30 min reoxygen-ation, while the cells in CON group were cultured un-der normal condition. After that, total RNA was ex-tracted and then subjected to miRNA microarray to screen differentially expressed miRNAs. The microar-ray results were further validated by quantitative RT-PCR ( qRT-PCR ) . Bioinformatics analysis was per-formed to predict the miRNAs-regulated target genes and analyze the enriched gene ontology ( GO) and sig-naling pathway ( Pathway) . Results HPC caused sig-nificant changes in miRNAs expression in cardiomyo-cytes as compared to CON group. A total of 12 miR-NAs were up-regulated and 14 miRNAs were down-reg-ulated ( P 500 were selected for further bioinformatics analysis. The expression of miR-133b-5p, miR-664-1-5p and miR-6216 detected by qRT-PCR exhibited the similar patterns of up or down regulation to those shown in mi-croarray results. Bioinformatics analysis revealed that miRNAs-regulated target genes were significantly en-riched in 27 GOs and 6 signal pathways. Conclusion The expression profile of miRNAs in rat cardiomyo-cytes is significantly affected by HPC. These differenti-ally expressed miRNAs might participate in HPC-in-duced cardioprotection by regulating their target genes in rat cardiomyocytes.

The protective effect of bone marrow mesenchymal stem cells by anoxic preconditioning on diabetic cardiomyopathy / 中华急诊医学杂志

Objective To explore the protective effect of bone marrow mesenchymal stem cells (MSC) on diabetic myocardium and anoxic pre-conditioning (AP).Methods Eight-week-- old male Sprague-Dawley rats were given with a single intraperitoneal injection of streptozotocin (STZ) (60 mg/kg)to induce diabetes mellitus (DM).Donor rats were 8-week-old male Sprague - Dawley rats.Before transplantation,MSC were incubated in CM-DiI at a concentration of 2 μg/mL for 20 min.AP-MSC were exposed to 3 hours of anoxia.At 4 months after STZ injection,diabetic rats were randomly given with an intramyocardial injection of one of the followings:150 μL of Dulbecco's modified Eagle's medium ( DMEM),5 ×106 MSC/150 μL,or 5 × 106 AP - MSC/150 μL (n =10 for each group).Three months after STZ injection and 2 weeks after transplantation,we evaluated the cardiac function by echocardiography,and also evaluated the cardiac conditions by alkaline phosphatase staining,western blot analysis for apoptosis related proteins and signal pathways.Results MSC,especially AP- MSC increased fractional shortening (FS) of diabetic heart (P ＜0.01 vs DMEM respectively).AP-MSC greatly increased the capillary density of diabetic myocardium (P ＜0.01 vs DMEM and MSC group respectively).AP-MSC are anti-apoptotic in the rat DCM model,possibly mediated through cardiac upregulation of Bcl-2/Bax ratio ( P ＜ 0.05 ) and inhibiting the expression and activation of caspase - 3 ( P ＜ 0.01 ).Conclusions Intramyocardial transplantation of APMSC has a protective effect on diabetic cardiomyopathy.

Gentamicin-induced preconditioning of proximal tubular LLC-PK1 cells stimulates nitric oxide production but not the synthesis of heat shock protein

Nephrotoxicity is the main side effect of antibiotics such as gentamicin. Preconditioning has been reported to protect against injuries as ischemia/reperfusion. The objective of the present study was to determine the effect of preconditioning with gentamicin on LLC-PK1 cells. Preconditioning was induced in LLC-PK1 cells by 24-h exposure to 2.0 mM gentamicin (G/IU). After 4 or 15 days of preconditioning, cells were again exposed to gentamicin (2.0 mM) and compared to untreated control or G/IU cells. Necrosis and apoptosis were assessed by acridine orange and HOESCHT 33346. Nitric oxide (NO) and endothelin-1 were assessed by the Griess method and available kit. Heat shock proteins were analyzed by Western blotting. After 15 days of preconditioning, LLC-PK1 cells exhibited a significant decrease in necrosis (23.5 ± 4.3 to 6.5 ± 0.3%) and apoptosis (23.5 ± 4.3 to 6.5 ± 2.1%) and an increase in cell proliferation compared to G/IU. NO (0.177 ± 0.05 to 0.368 ± 0.073 ìg/mg protein) and endothelin-1 (1.88 ± 0.47 to 2.75 ± 0.53 pg/mL) production significantly increased after 15 days of preconditioning compared toG/IU. No difference in inducible HSP 70, constitutive HSC 70 or HSP 90 synthesis in tubular cells was observed afterpreconditioning with gentamicin. The present data suggest that preconditioning with gentamicin has protective effects on proximal tubular cells, that involved NO synthesis but not reduction of endothelin-1 or production of HSP 70, HSC 70, or HSP 90. We conclude that preconditioning could be a useful tool to prevent the nephrotoxicity induced by gentamicin.

Pérdida del precondicionamiento isquémico: ¿un fenómeno relacionado con el envejecimiento? / Loss of Ischemic Preconditioning: An Aging Related Phenomenon

Introducción La edad es un predictor independiente de riesgo en pacientes con enfermedad coronaria. Esto podría explicarse por la falta de adaptación a la isquemia miocárdica aguda. El precondicionamiento es un mecanismo por el cual episodios repetitivos de isquemia inducen en el miocardio una tolerancia mayor a episodios subsiguientes. Objetivo Evaluar el desarrollo de precondicionamiento isquémico en pacientes añosos. Material y métodos Se incluyeron 65 pacientes sometidos a angioplastia coronaria electiva (< 70 años [n = 47] y ≥ 70 años [n = 18]). Se evaluó el desarrollo de precondicionamiento durante tres períodos de oclusión coronaria. Por ECG intracoronario se midió la elevación del ST al final de cada dilatación y se registró el porcentaje de resolución del ST a la tercera dilatación respecto del máximo valor registrado. Los datos se presentan como mediana e intervalo intercuartil 25/75%. Resultados No hubo diferencias significativas en las características clínicas basales. El máximo ST registrado, el ST a la tercera dilatación y el porcentaje de resolución del ST fueron 14 (9/24) mm, 8 (4/14) mm y 23,8% (0/55,5) para los pacientes jóvenes y 9,5 (5/18) mm (p = ns), 6,5 (4/ 16) mm (p = ns) y 5,5% (0/20) (p = 0,04) para los añosos. Al estratificar por grupos etarios, la proporción de pacientes que alcanzaron una resolución del ST ≥ 50% mostró una distribución lineal por chi cuadrado de tendencia (p = 0,025). Conclusiones Nuestro estudio sugiere que el precondicionamiento isquémico se encontraría disminuido en pacientes añosos. La funcionalidad de este mecanismo menguaría en forma progresiva con el envejecimiento.

Introduction Age is an independent risk predictor in patients with coronary disease. This could be explained by the lack of adaptation to acute myocardial ischemia. Preconditioning is a mechanism whereby repeated ischemia episodes induce in the myocardium an increased tolerance to further episodes. Objective To assess the development of ischemic preconditioning in elderly patients. Material and methods Sixty five patients who underwent elective coronary angioplasty were enrolled (<70 years [n=47] and ³70 years [n=18]). Preconditioning development was assessed during three periods of coronary occlusion. ST elevation at the end of each dilation was measured by intra coronary EKG, and ST resolution percentage after the third dilation was recorded and compared to the maximum recorded value. Data are presented as mean and inter-quartile interval 25/75%. Results There were no significant differences in the clinical baseline characteristics. The maximum ST recorded, ST after the third dilation and ST resolution percentage were 14 (9/24) mm, 8 (4/14) mm y 23.8% (0/55.5) for the young patients and 9.5 (5/ 18) mm (p = ns), 6.5 (4/16) mm (p=ns) and 5.5% (0/20) (p=0.04) for the elderly. Upon stratification per age group, the ration of patients that reached an ST resolution ≥ 50% showed a linear distribution by chi square for trend (p=0.025). Conclusions Our study suggests that ischemic preconditioning would be decreased in elderly patients. The functionality of this mechanism would progressively decrease with aging.

Teste de tetrazólio para avaliação da qualidade fisiológica de sementes de melancia / Tetrazolium test in the evaluation of watermelon physiological seed quality

O teste de tetrazólio é um dos métodos mais promissores para estimar, de forma rápida, a viabilidade e ovigor das sementes. Sua utilização em sementes de melancia requer cuidados na embebição e manuseiopela presença de camada mucilaginosa aderente ao tegumento. O objetivo do trabalho foi definir o tempode embebição e do método de escarificação para retirada da mucilagem, no pré-condicionamento doteste de tetrazólio, sementes de diferentes cultivares de melancia. As sementes foram imersas em água a30oC por 12 e 18 horas, e escarificadas em cal, areia fina e areia grossa. Após corte longitudinal na porçãodistal ao eixo embrionário, os tegumentos foram retirados e os embriões permaneceram em água por mais2 horas a 30oC para remoção manual da membrana remanescente. Após definição do método ideal de précondicionamento,os embriões foram imersos em soluções de tetrazólio nas concentrações de 0,075%;0,5% e 1% por três e quatro horas a 30oC. A imersão das sementes de melancia em água a 30oC por 12horas, retirada da mucilagem com areia fina, corte longitudinal na porção distal ao eixo embrionário eremoção do tegumento, seguido da permanência dos embriões em água por 2 horas a 30oC para retiradamanual da membrana interna e imersão na solução de tetrazólio a 0,075% por 4h a 30oC são procedimentosadequados para avaliação da qualidade de sementes de melancia.

The tetrazolium test is one of the most promising methods to estimate in a fast way viability and vigor of seeds. Its usage in watermelon seeds requires some care with the imbibition and handling due to presence of mucilaginous layer adherent to the tegument. The objective of this research work was to define the imbibition time and the method of scarification for removing the mucilage, during pre – conditioning of the tetrazolium test in watermelon. Seeds from different watermellon cultivars were immersed in water at 30°C from 12 to 18 hours, and sloughed in cal, fine and thick sand. After longitudinal cut in the distal portion to the embryonic axis, the tegument was removed and embryos were kept in water for additonal 2 hours at 30°C for hand removal of the remaining membrane. After definition of the ideal pre – conditioning method, the embryos were immersed in tetrazolium solution in concentrations of 0,075%; 0, 5% and 1% for 3 and 4 hours at 30°C. The immersion of watermelon seeds in water at 30°C for 12 hours, mucilage removal with fine sand, longitudinal cut in the distal portion to the embryonic axis, and removal of tegument, followed by the permanence of embryos in water for 2 hours at 30°C for the hand removal of the internal membrane and immersion in the tetrazolium solution at 0,075% for 4 hours at 30°C are adequate procedures for the evaluation of watermelon seed quality.

Nonmyeloablative allogeneic hematopoietic stem-cell transplantation a new immunotherapy for solid tumor / 医学研究生学报

Based on the practive of traditional allogeneic hematopoietic stem-cell transplantation therapy allo-HSCT and the development of nonmyeloablative conditioning, a new immunotherapy for solid tumor nonmyeloablative allo-HSCT, has been initiated. The nonmyeloablative,preparative regimens are associated with the use of lower dose of drugs,and less treatment-related toxicities. Instead of achieving maximal tumor reduction, the regimens are disigned to induce adequate immunosuppression to permit the engraftment of donor hematopoietic stem cells and serve as the platform for the administration of donor T cell in adoptive cell therapy. Donor's T-cell mediates a graft-versus-tumor(GVT)effect. This response is effective for the eradication of acceptor's tumor cell. This article reviews the concept, rationale, early clinical results, and limitation of nonmyeloablative allo-HSCT as a novel immunotherapy in solid tumors.