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Braz. j. med. biol. res ; 53(2): e8616, 2020. graf
Article in English | LILACS | ID: biblio-1055497


Previous research has shown that suppression of miR-383 can prevent inflammation of the endothelium, as well as postpone the development of atherosclerosis. However, the role of miR-383 in endothelial cell apoptosis in diabetes remains unclear. The aim of this study was to investigate the function of miR-383 in high glucose-induced apoptosis and oxidative stress in endothelial cells. A series of experiments involving qualitative polymerase chain reaction, cell transfection, luciferase assay, assessment of cell death, detection of catalase and superoxide dismutase concentrations, detection of intracellular reactive oxygen species (ROS), and western blot analysis were performed in this study. We found that miR-383 expression was promoted, while NAD+-dependent deacetylase and sirtuin 1 (SIRT1) expressions were suppressed in the endothelium of the aorta in db/db mice as well as in human umbilical vein endothelial cells, which were treated with high glucose (HG). Increased expression of miR-383 decreased expression of SIRT1, while suppression of miR-383 promoted expression of SIRT1 in human umbilical vein endothelial cells (HUVECs). Furthermore, suppression of miR-383 following transfection with miR-383 suppressor repressed cell death and generation of ROS in HUVECs. SIRT1 knockdown by siRNA-SIRT1 reversed the suppressive effect of miR-383 inhibition on ROS production and cell apoptosis induced by HG treatment. Overall, the findings of our research suggested that suppression of miR-383 repressed oxidative stress and reinforced the activity of endothelial cells by upregulation of SIRT1 in db/db mice, and targeting miR-383 might be promising for effective treatment of diabetes.

Animals , Male , Rabbits , Endothelium, Vascular/drug effects , Apoptosis/drug effects , Oxidative Stress/drug effects , MicroRNAs/antagonists & inhibitors , Sirtuin 1/metabolism , Glucose/pharmacology , Endothelium, Vascular/cytology , Endothelium, Vascular/physiology , Signal Transduction , Cells, Cultured , Mice, Inbred C57BL
Acta cir. bras ; 34(4): e201900402, 2019. tab, graf
Article in English | LILACS | ID: biblio-1001091


Abstract Purpose: To evaluate the effect of amniotic fluid in liver preservation in organ transplantation, and compare it with standard preservation solutions. Methods: The groups consisted of Group 1: Ringer Lactate (RL) group, Group 2: HTK group, Group 3: UW group, Group 4: AF group. The livers of rats from Group 1, 2, 3, and 4 were perfused and placed into falcon tubes containing RL, HTK, UW, and AF solutions at +4‎°C, respectively. The tubes were stored for 12 hours in the refrigerator at +4°C. Tissue samples were taken at the 6th and 12th hours for histopathological examinations of the perfused livers, and storage solutions for biochemical analyzes at 6th and 12th hours. Results: AF was shown to maintain organ viability by reducing the number of cells undergoing apoptosis. Histopathological changes such as sinusoidal dilatation, hydropic degeneration, and focal necrosis were found to be similar to the groups in which the standard organ preservation solutions were used. Additionally, the results of INOS, IL-10, and TNF-α,which were evaluated immunohistochemically, have been shown to be similar to the UW and HTK groups. Conclusions: AF provided conservation similar to UW and HTK in the 12-hour liver SCS process. The fact that apoptosis values are comparable to standard preservation solutions supports the success of AF in the cold storage of the liver.

Animals , Male , Cryopreservation/methods , Organ Preservation Solutions/pharmacology , Amniotic Fluid , Liver/blood supply , Liver/pathology , Organ Preservation/methods , Potassium Chloride/pharmacology , Procaine/pharmacology , Reference Values , Time Factors , Tissue Survival , Immunohistochemistry , Reperfusion Injury/prevention & control , Random Allocation , Reproducibility of Results , Tumor Necrosis Factor-alpha/analysis , Interleukin-10/analysis , Rats, Wistar , In Situ Nick-End Labeling , Nitric Oxide Synthase Type II/analysis , Ringer's Solution/pharmacology , Glucose/pharmacology , Mannitol/pharmacology
Rev. Soc. Bras. Med. Trop ; 52: e20180182, 2019. tab, graf
Article in English | LILACS | ID: biblio-1041508


Abstract INTRODUCTION: Administration of total parenteral nutrition (TPN) via catheters increases the risk for candidemia from Candida parapsilosis. METHODS: C. parapsilosis sensu stricto blood isolates were evaluated for ability total biomass biofilm formation and morphogenesis in presence of glucose at TPN equivalent concentrations. RESULTS: Biofilms were increased at high glucose concentrations (25-30%) compared to the control medium. Significant increase in filamentous forms was observed in cultures with 30% glucose. CONCLUSIONS: Biofilm formation by C. parapsilosis sensu stricto in hyperglycidic medium may contribute to its pathogenic potential for fungemia related to TPN catheters.

Humans , Biofilms/growth & development , Candida parapsilosis/physiology , Glucose/pharmacology , Colony Count, Microbial , Parenteral Nutrition, Home Total , Biofilms/drug effects , Culture Media/chemistry
J. bras. pneumol ; 45(4): e20170288, 2019. graf
Article in English | LILACS | ID: biblio-1040273


ABSTRACT Liquid perfluorocarbon (PFC) instillation has been studied experimentally as an adjuvant therapy in the preservation of lung grafts during cold ischemia. The objective of this study was to evaluate whether vaporized PFC is also protective of lung grafts at different cold ischemia times. We performed histological analysis of and measured oxidative stress in the lungs of animals that received only preservation solution with low-potassium dextran (LPD) or vaporized PFC together with LPD. We conclude that vaporized PFC reduces the production of free radicals and the number of pulmonary structural changes resulting from cold ischemia.

RESUMO O perfluorocarbono (PFC) líquido tem sido estudado experimentalmente como uma substância adjuvante na preservação de enxertos pulmonares durante o período de isquemia fria. O objetivo deste estudo foi avaliar se o PFC vaporizado (e não instilado) também atuaria como protetor de enxertos pulmonares em diferentes tempos de isquemia fria. Realizamos análise histológica e dosamos o estresse oxidativo em pulmões de animais que receberam somente uma solução de preservação com low-potassium dextran (LPD, dextrana com baixa concentração de potássio) ou PFC vaporizado associado a LPD. Concluímos que o PFC vaporizado reduziu a produção de radicais livres e provocou menor número de alterações estruturais pulmonares decorrentes do período de isquemia fria que o uso de LPD isoladamente.

Humans , Organ Preservation/methods , Lung Transplantation/methods , Oxidative Stress/drug effects , Cold Ischemia/methods , Fluorocarbons/pharmacology , Lung/drug effects , Reference Values , Time Factors , Reproducibility of Results , Dextrans/pharmacology , Organ Preservation Solutions , Glucose/pharmacology , Lung/pathology
Rev. bras. cir. cardiovasc ; 33(3): 211-216, May-June 2018. tab, graf
Article in English | LILACS | ID: biblio-958403


Abstract Objective: The present study aimed the functional recovery evaluation after long term of cardiac arrest induced by Custodiol (crystalloid-based) versus del Nido (blood-based) solutions, both added lidocaine and pinacidil as cardioplegic agents. Experiments were performed in isolated rat heart perfusion models. Methods: Male rat heart perfusions, according to Langendorff technique, were induced to cause 3 hours of cardiac arrest with a single dose. The hearts were assigned to one of the following three groups: (I) control; (II) Custodiol-LP; and (III) del Nido-LP. They were evaluated after ischemia throughout 90 minutes of reperfusion. Left ventricular contractility function was reported as percentage of recovery, expressed by developed pressure, maximum dP/dt, minimum dP/dt, and rate pressure product variables. In addition, coronary resistance and myocardial injury marker by alpha-fodrin degradation were also evaluated. Results: At 90 minutes of reperfusion, both solutions had superior left ventricular contractile recovery function than the control group. Del Nido-LP was superior to Custodiol-LP in maximum dP/dt (46%±8 vs. 67%±7, P<0.05) and minimum dP/dt (31%±4 vs. 51%±9, P<0.05) variables. Coronary resistance was lower in del Nido-LP group than in Custodiol-LP (395%±50 vs. 307%±13, P<0.05), as well as alpha-fodrin degradation, with lower levels in del Nido-LP group (P<0.05). Conclusion: Del Nido-LP cardioplegia showed higher functional recovery after 3 hours of ischemia. The analysis of alpha-fodrin degradation showed del Nido-LP solution provided greater protection against myocardial ischemia and reperfusion (IR) in this experimental model.

Animals , Male , Cardioplegic Solutions/pharmacology , Myocardial Reperfusion/methods , Potassium Compounds/pharmacology , Pinacidil/pharmacology , Heart Arrest, Induced/methods , Lidocaine/pharmacology , Time Factors , Vascular Resistance/physiology , Cardioplegic Solutions/chemistry , Carrier Proteins/analysis , Blotting, Western , Rats, Wistar , Coronary Vessels/physiopathology , Glucose/pharmacology , Glucose/chemistry , Heart/drug effects , Mannitol/pharmacology , Mannitol/chemistry , Microfilament Proteins/analysis
Braz. j. med. biol. res ; 51(1): e6378, 2018. tab
Article in English | LILACS | ID: biblio-889003


The aim of this study was to discuss the safety and efficacy of regional citrate anticoagulation (RCA) on continuous blood purification (CBP) during the treatment of multiple organ dysfunction syndrome (MODS). Thirty-five patients with MODS were divided into two groups: the local citrate anticoagulation (RCA) group, and the heparin-free blood purification (hfBP) group. The MODS severity was assessed according to Marshall's MODS score criteria. Blood coagulation indicators, blood pressure, filter lifespan, filter replacement frequency, anticoagulation indicators, and main metabolic and electrolyte indicators were analyzed and compared between RCA and hfBP groups. RCA resulted in lower blood pressure than hfBP. The filter efficacy in RCA treatment was longer than in the hfBP group. The blood clearance of creatine, blood urea nitrogen and uric acid was better in the RCA group. RCA also led to higher pH than hfBP. Neither treatment resulted in severe bleeding events. In addition, MODS score was positively correlated with prothrombin time and activated partial thromboplastin time but negatively correlated with platelet concentration. RCA is a safer and more effective method in CBP treatment; however, it could also lead to low blood pressure and blood alkalosis.

Humans , Male , Female , Adult , Middle Aged , Aged , Hemofiltration/methods , Citrates/pharmacology , Citric Acid/pharmacology , Glucose/pharmacology , Multiple Organ Failure/therapy , Anticoagulants/pharmacology , Reference Values , Severity of Illness Index , Blood Coagulation/drug effects , Heparin/pharmacology , Reproducibility of Results , Treatment Outcome , Anticoagulants/therapeutic use
São Paulo; s.n; 2018. 108 p.
Thesis in Portuguese | LILACS | ID: biblio-916041


Introdução: O diabetes mellitus (DM) está associado a complicações que comprometem a qualidade de vida e a sobrevida dos indivíduos. Além disso, acarreta elevados custos para o controle metabólico e o tratamento de suas complicações, sendo assim caracterizado como um problema de saúde pública. A regulação da digestão e da absorção intestinal dos carboidratos, com vista a manter a homeostase da glicose plasmática, constituem importantes estratégias de proteção em condições clínicas como o diabetes tipo 2 (DM2), obesidade e síndrome metabólica. Os compostos fenólicos compreendem um grupo complexo de fitoquímicos bioativos presentes nos vegetais. Estudos in vitro e in vivo têm demonstrado que os compostos fenólicos inibem a atividade de carbohidrases (α-amilase e α-glicosidase) e o transporte intestinal de glicose mediado pelos transportadores SGLT1 e GLUT2. O cerrado brasileiro compreende uma larga biodiversidade, porém, apesar de muitas espécies terem sido identificadas, o seu potencial nutritivo e funcional ainda é pouco conhecido. Dentre estas espécies nativas é destacado o jatobá-do-cerrado. O jatobá-do-cerrado é uma leguminosa nativa brasileira, cuja a polpa farinácea que envolve suas sementes apresenta quantidades significativas de compostos fenólicos, podendo ter um potencial efeito sobre o metabolismo da glicose. Objetivos: Verificar os efeitos dos compostos fenólicos da farinha de jatobá-do-cerrado na digestão de carboidratos e na captação de glicose em células intestinais Caco-2. Metodologia: Os compostos fenólicos da farinha de jatobá foram obtidos por extração sequencial com as soluções de etanol (60%) e acetona (70%). Em seguida, o extrato foi digerido utilizando enzimas (α-amilase, pepsina e pancreatina) em pH fisiológico. Os compostos fenólicos presentes no extrato antes e após a digestão foram identificados por cromatografia líquida de ultra performance - espectrômetro de massas (UPLC-MS/MS). Foi avaliada a capacidade de inibição dos extratos de jatobá digeridos em relação à atividade das enzimas α-amilase e α-glicosidase. Células intestinais Caco-2 foram incubadas com diferentes concentrações (0,05 mg/mL - 0,1 mg/mL) de extratos de farinha de jatobá digeridos em diferentes tempos (30 min, 2h e 12 h) para a avaliação da captação de glicose e da expressão gênica dos transportadores de glicose SGLT1 e GLUT2. Resultados: 44 compostos fenólicos foram identificados, dentre eles, a principal classe presente são os flavonoides. Compostos como o ácido cafeico, o kaempferol, quercetina-3- rutinosideo e a quercetrina estavam presentes no extrato antes da digestão. O conteúdo de compostos fenólicos do extrato foi reduzido após a digestão, entretanto o mesmo ainda apresentou compostos de relevância biológica como o ácido p-cumárico, ácido 3-o-feruloilquinico, theaflavina, crisina e grandinina que já apresentaram efeito positivo sobre o metabolismo da glicose in vitro em outros trabalhos. Os extratos fenólicos de jatobá após a digestão in vitro inibiram significativamente a atividade das enzimas α-amilase (76 e 91%) e α- glicosidase (53 e 77%). Os extratos também demonstraram inibir significativamente tanto a captação de glicose independente de sódio quanto a expressão gênica dos transportadores de glicose SGLT1 e GLUT2 de maneira dose-dependente. Conclusão: Este é o primeiro trabalho que identificou os compostos fenólicos presentes na farinha de jatobá. A partir do exposto, podemos concluir que a farinha de jatobá apresenta potencial benefício a saúde devido ao seu conteúdo de compostos fenólicos e a capacidade destes compostos de regular a digestão e a absorção de carboidratos in vitro

Introduction: Diabetes mellitus (DM) is associated with complications that decrease the quality of life and survival of individuals. In addition, it entails high costs for metabolic control and treatment of its complications, thus being characterized as a public health problem. The regulation of digestion and intestinal absorption of carbohydrates to maintain plasma glucose homeostasis are important strategies for protection in chronic diseases such as type 2 diabetes (DM2), obesity and metabolic syndrome. Phenolic compounds are a complex group of chemical substances present in plants. In vitro and in vivo studies have shown that phenolic compounds are able to inhibit the activity of carbohydrases (α-amylase and α-glycosidase) and the intestinal transport of glucose mediated by SGLT1 and GLUT2 transporters. Brazilian Cerrado present a large biodiversity, but although many species have been identified, its nutritional and functional potential is still little known. Among these native species is the jatobá-docerrado. Jatobá-do-cerrado is a brazilian native legume, whose farinaceous pulp that surrounds its seeds presents significant amounts of phenolic compounds and may have a potential effect on glucose metabolism. Objectives: To verify the effects of phenolic compounds from jatobá-do-cerrado flour in the digestion of carbohydrates and uptake of glucose in Caco-2 intestinal cells. Methods: Phenolic compounds of jatobá flour were obtained by sequential extraction with olutions of ethanol (60%) and acetone (70%). The extract was digested using enzymes (α-amylase, pepsin and pancreatin) at physiological pH. The phenolic compounds present in the extract before and after the digestion were identified by liquid chromatography of ultra-performance - mass spectrometer (UPLCMS / MS). The ability of inhibition of the extracts of jatobá digested in relation to the activity of α-amylase and α-glycosidase enzymes was evaluated. Caco-2 intestinal cells were incubated with different concentrations of jatobá flour extracts (0.1 mg / mL - 0.05 mg / mL) for different time (30 min, 2 h and 12 h) to the evaluation of facilitated uptake (sodium-free buffer) and gene expression of SGLT1 and GLUT2 glucose transporters. Results: 44 phenolic compounds have been identified, among them a major class present are flavonoids. Compounds such as caffeic acid, quercetin-3-rutinoside and quercetrine were present in the extract before in vitro digestion. The content of phenolic compounds of the extract after digestion was reduced. However, the extract presents compounds with biological activity such as p-coumaric acid, 3-o-feruloylquinic acid , theaflavin, chrysin and grandinine, which already presented positive effects on glucose metabolism in vitro in other studies. Phenolic extracts of jatobá after in vitro digestion inhibited the activity of α-amylase (76 and 91%) and α-glycosidase (53 and 77%). The extracts also shown to inhibit both glucose uptake and gene expression of glucose transporters SGLT1 and GLUT2 in a dose-dependent manner. Conclusion: This is the first work that identified the phenolic compounds present in jatobá flour. Thus, we can conclude that the jatobá flour presents potential health benefit by modulate digestion and the absorption of carbohydrates in vitro

Absorption, Physiological/genetics , Caco-2 Cells , Diabetes Mellitus , Fabaceae , Glucose/pharmacology , Phenolic Compounds , Biological Availability , Biological Transport , Digestion , Flour
Biol. Res ; 51: 22, 2018. graf
Article in English | LILACS | ID: biblio-950906


BACKGROUND: Our study aimed to investigate the roles of autophagy against high glucose induced response in retinal pigment epithelium (ARPE-19 cells). METHODS: The morphological changes and reactive oxygen species (ROS) generation in ARPE-19 cells under high glucose treatment were respectively detected using the transmission electron microscopy and flow cytometry. The expression levels of Parkin, PINK1, BNIP3L, LC3-I and LC3-II in ARPE-19 cells received high glucose treatment were measured by western blot after pretreatment of carbonyl cyanide m-chlorophenylhydrazone (CCCP), 3-methyladenine (3-MA), N-acetyl cysteine (NAC) or cyclosporin A (CsA) followed by high glucose treatment. RESULTS: ARPE-19 cells subjected to high glucose stress showed an obvious reduction in the LC3-I expression and significant increase in the number of autophagosomes, in the intracellular ROS level, and in the expression levels of Parkin, PINK1, BNIP3L and LC3-II (p < 0.05). Pretreatment with CCCP significantly reduced the LC3-I expression and increased the expression levels of Parkin, PINK1, BNIP3L and LC3-II (p < 0.05). ARPE-19 cells pretreated with CsA under high glucose stress showed markedly down-regulated expressions of Parkin, PINK1 and BNIP3L compared with the cells treated with high glucose (p < 0.05). Pretreatment of ARPE-19 cells with NAC or 3-MA under high glucose stress resulted in a marked reduction in the expression levels of PINK1, BNIP3L and LC3-II (p < 0.05). Meanwhile, the expression level of Parkin in the ARPE-19 cells pretreated with NAC under high glucose stress was comparable with that in the control cells. CONCLUSION: Autophagy might have protective roles against high glucose induced injury in ARPE19 cells via regulating PINK1/Parkin pathway and BNIP3L.

Humans , Protein Kinases/drug effects , Autophagy/drug effects , Proto-Oncogene Proteins/drug effects , Tumor Suppressor Proteins/drug effects , Ubiquitin-Protein Ligases/drug effects , Retinal Pigment Epithelium/drug effects , Glucose/pharmacology , Membrane Proteins/drug effects , Protein Kinases/metabolism , Autophagy/physiology , Signal Transduction/physiology , Cell Line , Proto-Oncogene Proteins/metabolism , Reactive Oxygen Species/metabolism , Tumor Suppressor Proteins/metabolism , Ubiquitin-Protein Ligases/metabolism , Microscopy, Electron, Transmission , Retinal Pigment Epithelium/cytology , Flow Cytometry , Membrane Proteins/metabolism
Mem. Inst. Oswaldo Cruz ; 112(9): 587-592, Sept. 2017. graf
Article in English | LILACS | ID: biblio-894875


BACKGROUND The link between Candida albicans and diabetes mellitus is well-acknowledged, but incompletely elucidated. OBJECTIVES The purpose of this study is to assess the growth rate of C. albicans (CA) in the presence of different concentrations of glucose and fructose, two of the main pathophysiologic and nutritionally relevant sugars in diabetic patients, in order to obtain a better understanding of the nutrient acquisition strategy and its possible relation to the hyperglycemic status of diabetic patients. METHODS The effects of different concentrations of glucose and fructose (1000 mg%, 500 mg%, 250 mg% and 100 mg% w/v) on the growth rate of CA have been studied by flow-cytometry. FINDINGS We found that glucose concentration is directly related to CA growth, which may be linked to the frequent yeast infections that occur in non-controlled diabetic patients; we also show that fructose inhibits CA growth rate. MAIN CONCLUSIONS As a consequence of our hypothesis, the study demonstrates that fructose-containing food may prevent the development of candidiasis, at least in oral sites.

Humans , Candida albicans/growth & development , Candida albicans/drug effects , Diabetes Mellitus/microbiology , Fructose/pharmacology , Glucose/pharmacology , Time Factors , In Vitro Techniques , Flow Cytometry
Rev. bras. enferm ; 70(2): 317-325, Mar.-Apr. 2017. tab
Article in English | LILACS, BDENF | ID: biblio-843639


ABSTRACT Objective: to assess pain in preterm newborns and to compare the neonatal and therapeutic variables with the total scores of the Neonatal Facial Coding System of preterm newborns submitted to arterial puncture exposed to music and 25% oral glucose. Method: a comparative study with 48 recordings of preterm newborns - Group 1, music (26); Group 2, glucose 25% (22) - individually analyzed by three trained nurses, after Kappa of at least 80%. Results: the variables and the pain scores of the groups did not present statistical significance (p < 0.05) according to the Neonatal Facial Coding System. 80.8% of the preterm infants in Group 1 had a higher quantitative score ≥ 3 in the neonatal variables (gender, type of delivery), and therapeutic variables (type of oxygen therapy, place of hospitalization, type of puncture). Conclusion: There was no difference when comparing the music and glucose 25% groups and the variables studied.

RESUMEN Objetivo: evaluar el dolor en recién nacidos prematuros y comparar las variables neonatales y terapéuticas con las puntuaciones totales del Neonatal Facial Coding System de los recién nacidos prematuros sometidos a una punción arterial expuestos a la música y glucosa al 25% por vía oral. Método: estudio comparativo con 48 fi lmaciones de los recién nacidos prematuros divididos en el Grupo 1 - música (26) y el Grupo 2 - glucosa al 25% (22). Las fi lmaciones fueron analizadas individualmente por tres enfermeras capacitadas después de coefi ciente Kappa de al menos 80%. Resultados: las variables y puntuaciones de dolor de los grupos no fueron estadísticamente signifi cativas (p<0,05) de acuerdo con el Neonatal Facial Coding System. En el Grupo 1, 80,8% de los recién nacidos prematuros mostraron mayores cantidades de puntuaciones ≥ 3 en las variables neonatales (sexo, tipo de parto) y las variables terapéuticas (tipo de la terapia de oxígeno, lugar de internación, tipo de punción). Conclusión: No hubo diferencias cuando se comparan los grupos de música y de glucosa al 25% y las variables estudiadas.

RESUMO Objetivo: avaliar a dor em recém-nascidos pré-termo e comparar as variáveis neonatais e terapêuticas com os escores totais da Neonatal Facial Coding System de recém-nascidos pré-termo submetidos à punção arterial exposto à música e glicose 25% oral. Método: estudo comparativo com 48 fi lmagens de recém-nascidos pré-termo - Grupo 1, música (26); Grupo 2, glicose 25% (22) - analisadas individualmente por três enfermeiras treinadas, após Kappa de no mínimo 80%. Resultados: as variáveis e os escores de dor dos grupos não apresentaram signifi cância estatística (p < 0,05) de acordo com o Neonatal Facial Coding System. 80,8% dos prematuros do Grupo 1 apresentaram um maior quantitativo de escores ≥ 3 nas variáveis neonatais (sexo, tipo de parto) e, variáveis terapêuticas (tipo de oxigenoterapia, local de internação, tipo de punção). Conclusão: Não houve diferença ao se comparar os grupos da música e da glicose 25% e as variáveis estudadas.

Humans , Male , Female , Infant, Newborn , Pain Measurement/methods , Needlestick Injuries/complications , Pain Management/standards , Pain/nursing , Acetylglucosamine/therapeutic use , Infant, Premature/psychology , Infant, Premature/blood , Pain Management/methods , Glucose/pharmacology , Music Therapy/methods , Music Therapy/standards
Braz. j. med. biol. res ; 49(4): e4324, 2016. tab, graf
Article in English | LILACS | ID: biblio-951663


The effects of interleukin-10 (IL-10) and glucose on mRNA and protein expression of osteoprotegerin (OPG), and its ligand, receptor activator of nuclear factor-κB ligand (RANKL), were investigated in human periodontal ligament fibroblasts (HPDLFs). Primary HPDLFs were treated with different concentrations of IL-10 (0, 1, 10, 25, 50, and 100 ng/mL) or glucose (0, 5.5, 10, 20, 30, and 40 mmol/L). Changes in mRNA and protein expression were examined using the reverse-transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. After IL-10 treatment, mRNA and protein levels of OPG were increased, while mRNA and protein levels of RANKL were decreased (P<0.05), both in a concentration-dependent manner. Glucose stimulation had the opposite concentration-dependent effect to that of IL-10 on OPG and RANKL expression. IL-10 upregulated OPG expression and downregulated RANKL expression, whereas high glucose upregulated RANKL and downregulated OPG in HDPLFs. Abnormal levels of IL-10 and glucose may contribute to the pathogenesis of periodontal disease.

Humans , Periodontal Ligament/drug effects , Interleukin-10/pharmacology , RANK Ligand/metabolism , Osteoprotegerin/metabolism , Fibroblasts/drug effects , Glucose/pharmacology , Periodontal Ligament/cytology , Time Factors , RNA, Messenger/analysis , Down-Regulation , Up-Regulation , Cells, Cultured , Blotting, Western , Analysis of Variance , Reverse Transcriptase Polymerase Chain Reaction , Fibroblasts/metabolism
Yonsei Medical Journal ; : 1252-1259, 2016.
Article in English | WPRIM | ID: wpr-79766


PURPOSE: Diabetic nephropathy (DN) is a prevalent chronic microvascular complication of diabetes mellitus involving disturbances in electrolytes and the acid-base balance caused by a disorder of glucose metabolism. NHE1 is a Na+/H+ exchanger responsible for keeping intracellular pH (pHi) balance and cell growth. Our study aimed to investigate roles of NHE1 in high glucose (HG)-induced apoptosis in renal tubular epithelial cells. MATERIALS AND METHODS: Renal epithelial tubular cell line HK-2 was cultured in medium containing 5 mM or 30 mM glucose. Then, cell apoptosis, oxidative stress, NHE1 expression, and pHi were evaluated. NHE1 siRNA and inhibitor were used to evaluate its role in cell apoptosis. RESULTS: HG significantly increased cell apoptosis and the production of reactive oxygen species (ROS) and 8-OHdG (p<0.05). Meanwhile, we found that HG induced the expression of NHE1 and increased the pHi from 7.0 to 7.6 after 48 h of incubation. However, inhibiting NHE1 using its specific siRNA or antagonist DMA markedly reduced cell apoptosis stimulated by HG. In addition, suppressing cellular oxidative stress using antioxidants, such as glutathione and N-acetyl cysteine, significantly reduced the production of ROS, accompanied by a decrease in NHE1. We also found that activated cyclic GMP-Dependent Protein Kinase Type I (PKG) signaling promoted the production of ROS, which contributed to the regulation of NHE1 functions. CONCLUSION: Our study indicated that HG activates PKG signaling and elevates the production of ROS, which was responsible for the induction of NHE1 expression and dysfunction, as well as subsequent cell apoptosis, in renal tubular epithelial cells.

Antioxidants/metabolism , Apoptosis/drug effects , Cation Transport Proteins/metabolism , Cell Cycle/drug effects , Cell Line , Dose-Response Relationship, Drug , Epithelial Cells/cytology , Glucose/pharmacology , Glutathione/metabolism , Humans , Kidney Tubules/cytology , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects , Sodium-Hydrogen Exchangers/metabolism
Acta cir. bras ; 30(1): 34-45, 01/2015. tab, graf
Article in English | LILACS | ID: lil-735704


PURPOSE: To evaluate the protective effects of chilling the bile ducts with cold (5°C) 5% glucose solution (GS) during radiofrequency (RF) administration. METHODS: Twenty male pigs (3 mos. old; 25-30 kg) were subjected to RF delivery with chilling (experimental group, N=10) or without chilling (control group, N=10). Half of the animals in each group were euthanized immediately after the operation, and half were euthanized one week later. The following histological variables in relation to the bile ducts were evaluated by a pathologist (blind examiner): degenerative changes to the epithelium; epithelial necrosis; ulceration, regenerative changes of the epithelium; polymorphonuclear neutrophil infiltration; and thermal effects. RESULTS: The experimental group (88 bile ducts examined) showed reduced thermal damage relative to the control group (86 bile ducts examined) as demonstrated by significant differences in the following histopathological parameters: epithelial detachment of biliary epithelium (84.1% vs. 59.3%; p<0.006); elongation/palisade arrangement of nuclei (65.1% vs. 87.5%; p<0.001); pseudo-goblet cells (32.9% vs. 56.8%; p<0.001). CONCLUSION: Infusion of 5% glucose solution (5°C) has a protective effect on bile ducts subjected to heat (95-110°C, 12 min) from radiofrequency thermal ablation device. .

Animals , Male , Bile Ducts/injuries , Burns/prevention & control , Catheter Ablation/adverse effects , Cryotherapy/methods , Glucose/pharmacology , Liver/surgery , Bile Duct Diseases/prevention & control , Bile Ducts/pathology , Burns/etiology , Catheter Ablation/methods , Hot Temperature/adverse effects , Perfusion , Protective Agents/pharmacology , Reproducibility of Results , Swine , Time Factors , Treatment Outcome
Rev. bras. enferm ; 67(6): 936-941, Nov-Dec/2014.
Article in Portuguese | LILACS, BDENF | ID: lil-732811


Pesquisa qualitativa, exploratória descritiva, que objetivou conhecer a percepção dos receptores sanguíneos quanto ao processo transfusional. A pesquisa foi realizada em uma unidade de hemoterapia de um município da região sul do Brasil e os dados foram analisados por meio do Discurso do Sujeito Coletivo. Foram entrevistados, por meio de instrumento semiestruturado, onze pacientes, homens e mulheres entre 30 e 95 anos, em recuperação pós-cirúrgica de cirurgia cardíaca, submetidos à transfusão sanguínea. Emergiram quatro Ideias Centrais: Perda e reposição sanguínea; Preservação da vida; Reconhecimento do processo transfusional e Segurança transfusional. A percepção sobre a mudança que os pós-transfundidos começam a vivenciar a partir do processo transfusional traz à tona uma ressignificação da própria vida. Este estudo mostrou que os pacientes transfundidos percebem o processo transfusional como uma alternativa de sobrevivência e, mesmo tendo conhecimento sobre o processo e seus significados, permanecem receios e angústias que podem ser minimizados pela equipe multiprofissional.

Qualitative research, descriptive exploratory, aimed to know the perception of blood transfusion recipients as to the process. The research was carried out at a blood bank in a city in southern Brazil, and the data were analyzed using the Collective Subject Discourse. Were interviewed using a semistructured instrument, eleven patients, men and women between 30 and 95 years, post-surgical recovery of cardiac surgery, underwent blood transfusion. Four central ideas emerged: loss and blood replacement; Preservation of life; Recognition of the transfusion process; and transfusion safety. The perception about the change that post-transfusion begin to live from the transfusion process raises a reframing of life itself. This study showed that transfused patients perceive the transfusion process as a means of survival, and even having knowledge about the process and their meanings, there is the permanence of fears and anxieties that can be minimized by the multidisciplinary team.

Investigación cualitativa, de tipo exploratorio descriptivo, con el objetivo de conocer la percepción de los receptores de transfusiones de sangre en cuanto al proceso. La investigación se llevó a cabo en un banco de sangre en una ciudad en el sur de Brasil, y los datos fueron analizados utilizando el Discurso del Sujeto Colectivo. Fueron entrevistados mediante un instrumento semi-estructurado, once pacientes, hombres y mujeres de entre 30 y 95 años de recuperación post-quirúrgica de la cirugía cardíaca, se sometió a una transfusión de sangre. Cuatro ideas centrales surgieron: la pérdida y reemplazo de sangre; La preservación de la vida; Reconocimiento del proceso de transfusión; y seguridad de las transfusiones. La percepción sobre el cambio que después de la transfusión comenzar a vivir desde el proceso de transfusión plantea una reformulación de la vida misma. Este estudio mostró que los pacientes transfundidos perciben el proceso de transfusión como un medio de supervivencia, e incluso tener conocimiento sobre el proceso y sus significados, no es la permanencia de los temores y ansiedades que pueden minimizarse por el equipo multidisciplinario.

Coenzyme A Ligases/biosynthesis , Phenylacetates/pharmacology , Pseudomonas/enzymology , Aerobiosis , Carbohydrate Metabolism , Chloramphenicol/pharmacology , Enzyme Induction , Glucose/metabolism , Glucose/pharmacology , Kinetics , Pseudomonas/drug effects , Pseudomonas/growth & development
Rev. bras. cir. cardiovasc ; 29(2): 156-162, Apr-Jun/2014. tab, graf
Article in Portuguese | LILACS | ID: lil-719409


Introdução: As soluções que provocam parada cardíaca eletiva estão em constante evolução, porém, o composto ideal ainda não foi encontrado. Os autores comparam uma nova solução cardioplégica com histidina-triptofano-glutamato (Grupo 2) com histidina-triptofano-cetoglutarato (Grupo 1) em modelo de coração isolado de rato. Objetivo: Quantificar a dimensão fractal e entropia de Shannon em miócitos de rato submetidos à cardioplegia utilizando solução histidina-triptofano com glutamato em modelo experimental, considerando-se os marcadores caspase, IL-8 e Ki-67. Métodos: Vinte ratos machos de raça Wistar foram anestesiados e heparinizados. O tórax foi aberto, realizado cardiectomia e infundido 40 ml/Kg de solução cardioplégica apropriada. Os corações foram mantidos por 2 horas na mesma solução a 4ºC e, após esse período, colocados em aparato de Langendorff por 30 minutos com solução de Ringer Locke. Foram feitas análises imunohistoquímicas para caspase, IL-8 e KI-67. Resultados: A dimensão fractal e a entropia de Shannon dos corações submetidos à parada cardíaca eletiva nos grupos 1 e 2 não foram diferentes. Conclusão: A quantidade de informações avaliada pela entropia de Shannon e a distribuição das mesmas (dada pela dimensão fractal) nas lâminas de coração de rato submetidas à cardioplegia com solução histidina-triptofano-acetoglutarato ou histidina-triptofano-glutamato não foram diferentes, o que mostra que a solução de histidina-triptofano-glutamato é tão boa quanto a histidina-triptofano-cetoglutarato na preservação dos miócitos em modelo de coração isolado de rato. .

Introduction: Solutions that cause elective cardiac arrest are constantly evolving, but the ideal compound has not yet been found. The authors compare a new cardioplegic solution with histidine-tryptophan-glutamate (Group 2) and other one with histidine-tryptophan-cetoglutarate (Group 1) in a model of isolated rat heart. Objective: To quantify the fractal dimension and Shannon entropy in rat myocytes subjected to cardioplegia solution using histidine-tryptophan with glutamate in an experimental model, considering the caspase markers, IL-8 and KI-67. Methods: Twenty male Wistar rats were anesthetized and heparinized. The chest was opened, the heart was withdrawn and 40 ml/kg of cardioplegia (with histidine-tryptophan-cetoglutarate or histidine-tryptophan-glutamate solution) was infused. The hearts were kept for 2 hours at 4ºC in the same solution, and thereafter placed in the Langendorff apparatus for 30 min with Ringer-Locke solution. Analyzes were performed for immunohistochemical caspase, IL-8 and KI-67. Results: The fractal dimension and Shannon entropy were not different between groups histidine-tryptophan-glutamate and histidine-tryptophan-acetoglutarate. Conclusion: The amount of information measured by Shannon entropy and the distribution thereof (given by fractal dimension) of the slices treated with histidine-tryptophan-cetoglutarate and histidine-tryptophan-glutamate were not different, showing that the histidine-tryptophan-glutamate solution is as good as histidine-tryptophan-acetoglutarate to preserve myocytes in isolated rat heart. .

Animals , Male , Cardioplegic Solutions/pharmacology , Glutamic Acid/pharmacology , Heart Arrest, Induced/methods , Myocytes, Cardiac/drug effects , Caspases/analysis , Disease Models, Animal , Entropy , Fractals , Glucose/pharmacology , Heart/drug effects , Immunohistochemistry , /analysis , /analysis , Mannitol/pharmacology , Potassium Chloride/pharmacology , Procaine/pharmacology , Rats, Wistar , Reproducibility of Results , Time Factors
Rev. bras. cir. cardiovasc ; 28(4): 524-530, out.-dez. 2013. ilus, tab
Article in English | LILACS | ID: lil-703122


INTRODUCTION: Myocardial preservation during open heart surgeries and harvesting for transplant are of great importance. The heart at the end of procedure has to resume its functions as soon as possible. All cardioplegic solutions are based on potassium for induction of cardioplegic arrest. OBJECTIVE: To assess a cardioplegic solution with no potassium addition to the formula with two other commercially available cardioplegic solutions. The comparative assessment was based on cytotoxicity, adenosine triphosphate myocardial preservation, and caspase 3 activity. The tested solution (LIRM) uses low doses of sodium channel blocker (lidocaine), potassium channel opener (cromakalin), and actin/myosin cross bridge inhibitor (2,3-butanedione monoxime). METHODS: Wistar rats underwent thoracotomy under mechanical ventilation and three different solutions were used for "in situ" perfusion for cardioplegic arrest induction: Custodiol (HTK), Braile (G/A), and LIRM solutions. After cardiac arrest, the hearts were excised and kept in cold storage for 4 hours. After this period, the hearts were assessed with optical light microscopy, myocardial ATP content and caspase 3 activity. All three solutions were evaluated for direct cytotoxicity with L929 and WEHI-164 cells. RESULTS: The ATP content was higher in the Custodiol group compared to two other solutions (P<0.05). The caspase activity was lower in the HTK group compared to LIRM and G/A solutions (P<0.01). The LIRM solution showed lower caspase activity compared to Braile solution (P<0.01). All solutions showed no cytotoxicity effect after 24 hours of cells exposure to cardioplegic solutions. CONCLUSION: Cardioplegia solutions without potassium are promised and aminoacid addition might be an interesting strategy. More evaluation is necessary for an optimal cardioplegic solution development.

INTRODUÇÃO: Preservação do miocárdio durante cirurgias cardíacas abertas e de colheita para transplante são de grande importância. O coração ao final do processo tem de retomar as suas funções, logo que possível. Todas as soluções cardioplégicas são baseadas em potássio, para indução de parada cardioplégica. OBJETIVO: Comparar a uma solução cardioplégica sem adição de potássio à sua fórmula com duas outras soluções cardioplégicas disponíveis comercialmente. A avaliação comparativa foi baseada na citotoxicidade, preservação miocárdica (adenosina trifosfato, ATP) e atividade da caspase 3. A solução testada (LIRM) utiliza baixas doses de bloqueador de canal de sódio (lidocaína), abridor do canal de potássio (cromacalina) e inibidor da ponte actina/miosina (2,3-butanodiona monoxima). MÉTODOS: Ratos Wistar foram submetidos à toracotomia sob ventilação mecânica e três soluções diferentes foram utilizadas para perfusão in situ para a indução de parada cardioplégica: soluções Custodiol (HTK) Braile (G/A) e LIRM. Após parada cardíaca, os corações foram retirados e mantidos em câmara fria por 4 horas. Após esse período, o coração foi avaliado com microscopia de luz ótica, o conteúdo de ATP miocárdico e atividade da caspase 3. Todas as três soluções foram avaliadas quanto à citotoxicidade direta com células L929 e WEHI-164. RESULTADOS: A quantidade de ATP foi maior no grupo Custodiol em comparação às com outras duas soluções (P<0,05). A atividade de caspase foi menor no grupo HTK quando comparado às soluções LIRM e G/A (P<0,01). A solução LIRM demonstrou menor atividade da caspase em comparação à solução Braile (P<0,01). Todas as soluções não mostraram qualquer efeito de citotoxicidade após 24 horas de exposição das células às soluções cardioplégicas. CONCLUSÃO: Soluções cardioplégicas sem potássio são uma perspectiva e a adição de aminoácido pode ser uma estratégia interessante. Mais avaliações são necessárias para o desenvolvimento ideal da solução cardioplégica.

Animals , Rats , Cardioplegic Solutions/pharmacology , Heart Arrest, Induced/methods , Heart/drug effects , Organ Preservation/methods , Adenosine Triphosphate/analysis , Cardioplegic Solutions/chemistry , /analysis , Cell Survival/drug effects , Glucose/chemistry , Glucose/pharmacology , Models, Animal , Mannitol/chemistry , Mannitol/pharmacology , Myocardial Reperfusion Injury/prevention & control , Potassium Chloride/chemistry , Potassium Chloride/pharmacology , Potassium/chemistry , Potassium/pharmacology , Procaine/chemistry , Procaine/pharmacology , Rats, Wistar , Reproducibility of Results , Sodium Channel Blockers/chemistry , Time Factors
Indian J Exp Biol ; 2013 Aug; 51(8): 615-622
Article in English | IMSEAR | ID: sea-149364


Oxazaphosphorines belong to a group of alkylating agents. Mafosfamide cyclohexylamine salt (D-17272), 4-hydro-peroxy-cyclophosphamide (D-18864) and glufosfamide (D-19575, β-D-glucose-isophosphoramide mustard) are new generation oxazaphosphorines. The objective of the present study was to compare the cytotoxic action of these oxazaphosphorine compounds against human histiocytic lymphoma U937 cells. The chemical structures of the oxazaphosphorines were responsible for the different responses of U937 cells. The cytotoxic effects of D-17272, D-18864, and D-19575 on U937 cells depended on the agent tested, its dose, and the time intervals after the oxazaphosphorine application. Among the oxazaphosphorine agents, D-18864 appeared to be the most cytotoxic, and D-19575 was characterized by the lowest cytotoxicity. The in vitro cytotoxic activities of the oxazaphosphorines were strongly associated with their cell death inducing potential.

Antineoplastic Agents, Alkylating/pharmacology , Apoptosis/drug effects , Cell Proliferation/drug effects , Cyclophosphamide/analogs & derivatives , Cyclophosphamide/pharmacology , Flow Cytometry , Glucose/analogs & derivatives , Glucose/pharmacology , Humans , Ifosfamide/analogs & derivatives , Ifosfamide/pharmacology , Lymphoma, Large B-Cell, Diffuse/drug therapy , Lymphoma, Large B-Cell, Diffuse/pathology , Membrane Potential, Mitochondrial/drug effects , Necrosis , Phosphoramide Mustards/pharmacology , Tumor Cells, Cultured
Clinics ; 68(2): 199-204, 2013. ilus, tab
Article in English | LILACS | ID: lil-668807


OBJECTIVE: Ophthalmologic examination for retinopathy of prematurity is a painful procedure. Pharmacological and non-pharmacological interventions have been proposed to reduce pain during eye examinations. This study aims to evaluate the analgesic effect of 25% glucose using a validated pain scale during the first eye examination for retinopathy of prematurity in preterm infants with birth weight <1,500 g and/or gestational age <32 weeks. METHODS: A masked, randomized clinical trial for one dose of 1 ml of oral 25% glucose solution 2 minutes before the first ophthalmologic examination for retinopathy of prematurity was conducted between March 2008 and April 2010. The results were compared to those of a control group that did not receive oral glucose solution. Pain was evaluated using a Neonatal Infant Pain Scale immediately before and immediately after the ophthalmologic examination in both groups. NCT00648687 RESULTS: One hundred and twenty-four patients who were examined for the first time for retinopathy of prematurity were included. Seventy were included in the intervention group and 54 in the control group. The number of patients with pain immediately before the procedure was similar in both groups. The number of patients with pain after ophthalmologic examination was 15.7% in the intervention group and 68.5% in the control group (p<0.001). CONCLUSIONS: One ml of oral 25% glucose solution given 2 minutes before an ophthalmologic examination for retinopathy of prematurity was an effective measure for pain relief.

Female , Humans , Infant, Newborn , Analgesics/administration & dosage , Eye Pain/prevention & control , Eye/drug effects , Glucose/administration & dosage , Retinopathy of Prematurity/diagnosis , Administration, Oral , Analysis of Variance , Analgesics/pharmacology , Glucose/pharmacology , Pain Measurement , Time Factors , Treatment Outcome
Article in English | WPRIM | ID: wpr-35842


The maintenance of whole-body glucose homeostasis is critical for survival, and is controlled by the coordination of multiple organs and endocrine systems. Pancreatic islet beta cells secrete insulin in response to nutrient stimuli, and insulin then travels through the circulation promoting glucose uptake into insulin-responsive tissues such as liver, skeletal muscle and adipose. Many of the genes identified in human genome-wide association studies of diabetic individuals are directly associated with beta cell survival and function, giving credence to the idea that beta-cell dysfunction is central to the development of type 2 diabetes. As such, investigations into the mechanisms by which beta cells sense glucose and secrete insulin in a regulated manner are a major focus of current diabetes research. In particular, recent discoveries of the detailed role and requirements for reorganization/remodeling of filamentous actin (F-actin) in the regulation of insulin release from the beta cell have appeared at the forefront of islet function research, having lapsed in prior years due to technical limitations. Recent advances in live-cell imaging and specialized reagents have revealed localized F-actin remodeling to be a requisite for the normal biphasic pattern of nutrient-stimulated insulin secretion. This review will provide an historical look at the emergent focus on the role of the actin cytoskeleton and its regulation of insulin secretion, leading up to the cutting-edge research in progress in the field today.

Actins/metabolism , Animals , Exocytosis/drug effects , Glucose/pharmacology , Humans , Insulin/metabolism , Insulin-Secreting Cells/drug effects , Signal Transduction/drug effects
J. bras. pneumol ; 38(4): 461-469, jul.-ago. 2012. ilus, tab
Article in Portuguese | LILACS | ID: lil-647812


OBJETIVO: Comparar os achados histopatológicos e de apoptose em pulmões de ratos preservados em soluções low-potassium dextran (LPD, baixo potássio dextrana), histidine-tryptophan-ketoglutarate (HTK, histidina-triptofano-cetoglutarato) ou salina normal (SN) em 6 h e 12 h de isquemia pela utilização de um modelo experimental de perfusão pulmonar ex vivo. MÉTODOS: Sessenta ratos Wistar foram anestesiados, randomizados e submetidos à perfusão anterógrada pela artéria pulmonar com uma das soluções preservadoras. Após a extração, os blocos cardiopulmonares foram preservados por 6 ou 12 h a 4ºC, sendo então reperfundidos com sangue homólogo em um sistema de perfusão ex vivo durante 60 min. Ao final da reperfusão, fragmentos do lobo médio foram extraídos e processados para histopatologia, sendo avaliados os seguintes parâmetros: congestão, edema alveolar, hemorragia alveolar, hemorragia, infiltrado inflamatório e infiltrado intersticial. O grau de apoptose foi avaliado pelo método TdT-mediated dUTP nick end labeling. RESULTADOS: A histopatologia demonstrou que todos os pulmões preservados com SN apresentaram edema alveolar após 12 h de isquemia. Não houve diferenças em relação ao grau de apoptose nos grupos estudados. CONCLUSÕES: No presente estudo, os achados histopatológicos e de apoptose foram semelhantes com o uso das soluções LPD e HTK, enquanto a presença de edema foi significativamente maior com o uso de SN.

OBJECTIVE: To compare histopathological findings and the degree of apoptosis among rat lungs preserved with low-potassium dextran (LPD) solution, histidine-tryptophan-ketoglutarate (HTK) solution, or normal saline (NS) at two ischemia periods (6 h and 12 h) using an experimental rat model of ex vivo lung perfusion. METHODS: Sixty Wistar rats were anesthetized, randomized, and submitted to antegrade perfusion via pulmonary artery with one of the preservation solutions. Following en bloc extraction, the heart-lung blocks were preserved for 6 h or 12 h at 4ºC and then reperfused with homologous blood for 60 min in an ex vivo lung perfusion system. At the end of the reperfusion, fragments of the middle lobe were extracted and processed for histopathological examination. The parameters evaluated were congestion, alveolar edema, alveolar hemorrhage, inflammatory infiltrate, and interstitial infiltrate. The degree of apoptosis was assessed using the TdT-mediated dUTP nick end labeling method. RESULTS: The histopathological examination showed that all of the lungs preserved with NS presented alveolar edema after 12 h of ischemia. There were no statistically significant differences among the groups in terms of the degree of apoptosis. CONCLUSIONS: In this study, the histopathological and apoptosis findings were similar with the use of either LPD or HTK solutions, whereas the occurrence of edema was significantly more common with the use of NS.

Animals , Male , Rats , Apoptosis , Lung , Liver/pathology , Organ Preservation Solutions/pharmacology , Organ Preservation/methods , Potassium Chloride/pharmacology , Glucose/pharmacology , Liver/drug effects , Mannitol/pharmacology , Pulmonary Edema , Perfusion/methods , Procaine/pharmacology , Random Allocation , Rats, Wistar