Renal lipotoxicity: Insights from experimental models.

In recent decades, there has been a progressive increase in the prevalence of obesity and chronic kidney disease. Renal lipotoxicity has been associated with obesity. Although lipids play fundamental physiological roles, the accumulation of lipids in kidney cells may cause dysfunction and/or renal fibrosis. Adipose tissue that exceeds their lipid storage capacity begins to release triglycerides into the bloodstream that can get stored in several organs, including the kidneys. The mechanisms underlying renal lipotoxicity involve intracellular lipid accumulation and organelle dysfunction, which trigger oxidative stress and inflammation that consequently result in insulin resistance and albuminuria. However, the specific pathways involved in renal lipotoxicity have not yet been fully understood. We aimed to summarize the current knowledge on the mechanisms by which lipotoxicity affects the renal morphology and function in experimental models of obesity. The accumulation of fatty acids in tubular cells has been described as the main mechanism of lipotoxicity; however, lipids and their metabolism also affect the function and the survival of podocytes. In this review, we presented indication of mitochondrial, lysosomal and endoplasmic reticulum alterations involved in kidney damage caused by obesity. The kidney is vulnerable to lipotoxicity, and studies of the mechanisms underlying renal injury caused by obesity can help identify therapeutic targets to control renal dysfunction.

Iron-based phosphorus chelator: Risk of iron deposition and action on bone metabolism in uremic rats.

Phosphate chelators are frequently used in patients with chronic kidney disease (CKD). New iron-based chelators remain understudied and offer a promising therapeutic option for the control of bone and mineral disorders of chronic kidney disease (BMD-CKD). We assessed the effect of the phosphorus chelator, chitosan-iron III (CH-FeCl), compared to calcium carbonate (CaCO3) in BMD-CKD and the potential iron overload in uremic rats. Thirty-two animals were divided into four groups, namely the control, CKD, CKD/CH-FeCl, and CKD/CaCO3 groups. CKD was induced by adding 0.75% (4 weeks) and 0.1% (3 weeks) adenine to the diet. The chelators were administered from week 3 through week 7. The renal function, BMD-CKD markers, and histomorphometry of the femur were assessed at week 7. The CKD group showed a significant increase in creatinine (83.9 ± 18.6 vs. 41.5 ± 22.1 µmol/L; P = 0.001), phosphate (3.5 ± 0.8 vs. 2.2 ± 0.2 mmol/L; P = 0.001), fractional excretion of phosphorus (FEP) (0.71 ± 0.2 vs. 0.2 ± 0.17; P = 0.0001), and FGF23 (81.36 ± 37.16 pg/mL vs. 7.42 ± 1.96; P = 0.011) compared to the control group. There was no accumulation of serum or bone iron after the use of CH-FeCl. The use of chelators reduced the FEP (control: 0.71 ± 0.20; CKD/CH-FeCl: 0.40 ± 0.16; CKD/CaCO3 0.34 ± 0.15; P = 0.001), without changes in the serum FGF23 and parathyroid hormone levels. Histomorphometry revealed the presence of bone disease with high remodeling in the uremic animals without changes with the use of chelators. The CH-FeCl chelator was efficient in reducing the FEP without iron accumulation, thereby paving the way for the use of this class of chelators in clinical settings in the future.

Management strategies for implementing a multicenter cross-sectional study: lessons from the ADHERE Brazil study.

BACKGROUND: Epidemiological studies involving large samples usually face financial and operational challenges. OBJECTIVES: To describe the planning and execution of ADHERE Brazil, an epidemiological study on 1,105 kidney transplant patients, and report on how the study was structured, difficulties faced and solutions found. DESIGN AND SETTING: Cross-sectional multicenter study in 20 Brazilian kidney transplantation centers. METHODS: Actions developed in each phase of implementation were described, with emphasis on innovations used within the logistics of this study, aimed at estimating the prevalence of nonadherence to treatment. RESULTS: Coordination of activities was divided into four areas: general, regulatory, data collection and statistics. Weekly meetings were held for action planning. The general coordination team was in charge of project elaboration, choice of participating centers, definition of publication policy and monitoring other coordination teams. The regulatory team provided support to centers for submitting the project to ethics committees. The data collection team prepared a manual on the electronic collection system, scheduled web meetings and was available to respond to queries. It also monitored the data quality and reported any inadequacies found. Communication with the centers was through monthly reports via e-mail and distribution of exclusive material. The statistical team acted in all phases of the study, especially in creating the data analysis plan and data bank, generation of randomization lists and data extraction. CONCLUSIONS: Through these logistics, we collected high-quality data and built a local research infrastructure for further studies. We present supporting alternatives for conducting similar studies. CLINICAL TRIAL ANNOTATION: http://clinicaltrials.gov/ on October 10, 2013; NCT02066935.

Public Knowledge and Attitudes Toward Organ Donation and Transplantation: A Survey From a Medium-Sized Brazilian City.

INTRODUCTION: In Brazil, where organ donation occurs after brain death (BD) and needs family consent, notification of potential donors increased in the last decade, although family refusal is still around 40%. Among modifiable factors, there is a lack of knowledge by the population regarding BD and the process of organ donation. Our goal was to evaluate the knowledge of the population about these topics in a medium-sized Brazilian city with a high family refusal rate. METHODS: In a cross-sectional study, we randomly recruited 868 individuals in public places in January through March 2017. We collected data about demographics and concepts related to organ donation by interview. Groups were compared by χ2 test. RESULTS: Most of the subjects were women (57.3%), 54.6% of whom were white, and the mean age was 37.1 ±15.8 years; 58.8% had at least 11 years of schooling, and 55.8% had a monthly familiar income of < 3 reference wages. Individuals who declared to be donors (67.5%) were younger (36.0 ± 14.9 vs 39.6 ± 17.4 years, P = .007) and had higher schooling (47.3 vs 28.7% with > 11 years of schooling, P < .001). Among donors, 41% did not inform their families, 38% believed that donation could occur regardless of the kind of death, and 26.8% believed in the reversibility of BD. At least 90% of individuals, however, independent from donation status, would authorize organ donation of a relative if they previously knew the donation status of that relative. CONCLUSION: We observed, despite a high prevalence of potential donors, conflictive concepts regarding BD. This information suggests directions for the design of educative measures.

Digital radiography as an alternative method in the evaluation of bone density in uremic rats.

INTRODUCTION: Digital radiography (DRx) may provide a suitable alternative to investigate mineral and bone disorder (MBD) and loss of bone density (BD) in rodent models of chronic kidney disease (CKD). The objective of this study was to use DRx to evaluate BD in CKD rats, and to evaluate the correlation between DRx findings and serum MBD markers and bone histomorphometry. METHODS: Uremia was induced by feeding Wistar rats an adenine-enriched diet (0.75% for 4 weeks/0.10% for 3 weeks); outcomes were compared to a control group at experimental weeks 3, 4, and 7. The following biochemical markers were measured: creatinine clearance (CrC), phosphate (P), calcium (Ca), fractional excretion of P (FeP), alkaline phosphatase (ALP), fibroblast growth factor-23 (FGF-23), and parathyroid hormone (PTH). DRx imaging was performed and histomorphometry analysis was conducted using the left femur. RESULTS: As expected, at week 7, uremic rats presented with reduced CrC and higher levels of P, FeP, and ALP compared to controls. DRx confirmed the lower BD in uremic animals (0.57±0.07 vs. 0.68 ± 0.06 a.u.; p = 0.016) compared to controls at the end of week 7, when MBD was more prominent. A severe form of high-turnover bone disease accompanied these biochemical changes. BD measured on DRx correlated to P (r=-0.81; p = 0.002), ALP (r = -0.69, p = 0.01), PTH (r = -0.83, p = 0.01), OS/BS (r = -0.70; p = 0.02), and ObS/BS (r = -0.70; p = 0.02). CONCLUSION: BD quantified by DRx was associated with the typical complications of MBD in CKD and showed to be viable in the evaluation of bone alterations in CKD.

Chitosan-Fe (III) Complex as a Phosphate Chelator in Uraemic Rats: A Novel Treatment Option.

Phosphate retention and hyperphosphataemia are associated with increased mortality in patients with chronic kidney disease (CKD). We tested the use of cross-linked iron chitosan III (CH-FeCl) as a potential phosphate chelator in rats with CKD. We evaluated 96 animals, divided equally into four groups (control, CKD, CH-FeCl and CKD/CH-FeCl), over 7 weeks. We induced CKD by feeding animals an adenine-enriched diet (0.75% in the first 4 weeks and 0.1% in the following 3 weeks). We administered 30 mg/kg daily of the test polymer, by gavage, from the third week until the end of the study. All animals received a diet supplemented with 1% phosphorus. Uraemia was confirmed by the increase in serum creatinine in week 4 (36.24 ± 18.56 versus 144.98 ± 22.1 µmol/L; p = 0.0001) and week 7 (41.55 ± 22.1 versus 83.98 ± 18.56 µmol/L; p = 0.001) in CKD animals. Rats from the CKD group treated with CH-FeCl had a 54.5% reduction in serum phosphate (6.10 ± 2.23 versus 2.78 ± 0.55 mmol/L) compared to a reduction of 25.6% in the untreated CKD group (4.75 ± 1.45 versus 3.52 ± 0.74 mmol/L, p = 0.021), between week 4 and week 7. At week 7, renal function in both CKD groups was similar (serum creatinine: 83.98 ± 18.56 versus 83.10 ± 23.87 µmol/L, p = 0.888); however, the CH-FeCl-treated rats had a reduction in phosphate overload measured by fractional phosphate excretion (FEPi) (0.71 ± 0.2 versus 0.4 ± 0.16, p = 0.006) compared to the untreated CKD group. Our study demonstrated that CH-FeCl had an efficient chelating action on phosphate.

Effect of cross-linked chitosan iron (III) on vascular calcification in uremic rats.

Cross-linked chitosan iron (III) is a chitin-derived polymer with a chelating effect on phosphorus, but it is untested in vascular calcification. We evaluated this compound's ability to reduce hyperphosphatemia and its effect on vascular calcification in uremic rats using an adenine-based, phosphorus-rich diet for seven weeks. We used a control group to characterize the uremia. Uremic rats were divided according the treatment into chronic kidney disease, CKD-Ch-Fe(III)CL (CKD-Ch), CKD-calcium carbonate, or CKD-sevelamer groups. We measured creatinine, phosphorus, calcium, alkaline phosphatase, phosphorus excretion fraction, parathyroid hormone, and fibroblast growth factor 23. Vascular calcification was assessed using the aortic calcium content, and a semi-quantitative analysis was performed using Von Kossa and hematoxylin-eosin staining. At week seven, rats in the chronic kidney disease group had higher creatinine, phosphorus, phosphorus excretion fraction, calcium, alkaline phosphatase, fibroblast growth factor 23, and aortic calcium content than those in the Control group. Treatments with cross-linked chitosan iron (III) and calcium carbonate prevented phosphorus increase (20%-30% reduction). The aortic calcium content was lowered by 88% and 85% in the CKD-Ch and CKD-sevelamer groups, respectively. The prevalence of vascular changes was higher in the chronic kidney disease and CKD-calcium carbonate (62.5%) groups than in the CKD-Ch group (37.5%). In conclusion, cross-linked chitosan iron (III) had a phosphorus chelating effect similar to calcium carbonate already available for clinical use, and prevented calcium accumulation in the aorta. Impact statement Vascular calcification (VC) is a common complication due to CKD-related bone and mineral disorder (BMD) and is characterized by deposition of calcium in vessels. Effective therapies are not yet available but new phosphorus chelators can prevent complications from CV. We tested the effect of chitosan, a new phosphorus chelator, on the VC of uremic animals. It has recently been proposed that chitosan treatment may be effective in the treatment of hyperphosphataemia. However, its action on vascular calcification has not been investigated yet. In this study, we demonstrated that chitosan reduced the calcium content in the aorta, suggesting that this may be a therapeutic approach in the treatment of hyperphosphatemia by preventing CV.

Management strategies for implementing a multicenter cross-sectional study: lessons from the ADHERE Brazil study

ABSTRACT BACKGROUND: Epidemiological studies involving large samples usually face financial and operational challenges. OBJECTIVES: To describe the planning and execution of ADHERE Brazil, an epidemiological study on 1,105 kidney transplant patients, and report on how the study was structured, difficulties faced and solutions found. DESIGN AND SETTING: Cross-sectional multicenter study in 20 Brazilian kidney transplantation centers. METHODS: Actions developed in each phase of implementation were described, with emphasis on innovations used within the logistics of this study, aimed at estimating the prevalence of nonadherence to treatment. RESULTS: Coordination of activities was divided into four areas: general, regulatory, data collection and statistics. Weekly meetings were held for action planning. The general coordination team was in charge of project elaboration, choice of participating centers, definition of publication policy and monitoring other coordination teams. The regulatory team provided support to centers for submitting the project to ethics committees. The data collection team prepared a manual on the electronic collection system, scheduled web meetings and was available to respond to queries. It also monitored the data quality and reported any inadequacies found. Communication with the centers was through monthly reports via e-mail and distribution of exclusive material. The statistical team acted in all phases of the study, especially in creating the data analysis plan and data bank, generation of randomization lists and data extraction. CONCLUSIONS: Through these logistics, we collected high-quality data and built a local research infrastructure for further studies. We present supporting alternatives for conducting similar studies. CLINICAL TRIAL ANNOTATION: http://clinicaltrials.gov/ on October 10, 2013; NCT02066935.

Standardization of renal function evaluation in Wistar rats (Rattus norvegicus) from the Federal University of Juiz de Fora's colony.

INTRODUCTION: There is great interest in the use of animal models in the study of renal pathophysiology requires standardization of parameters. OBJECTIVE: Standardize assessment of renal function in rats from in the Center for Reproductive Biology of Federal University of Juiz de Fora's colony. METHODS: Thirty Wistar rats were used and performed measurements of creatinine (serum and urine), serum urea and proteinuria. Were evaluated: the urine collection interval in metabolic cages (24 hours or 12 hours), the need for 12-hour fast, the need of urine and serum deproteinization for creatinine measurement, need of serum deproteinization in animals with acute kidney injury to a spectrophotometer and ELISA, and the comparison of 24-hour proteinuria (PT 24 hours) with the protein/creatinine ratio (rP/C). Means were compared by the Student's t test, Pearson correlation, Bland-Altman plot for agreement and linear regression model to estimate PT 24 hours from rP/C. RESULTS: The 24 hours urine output was greater than 12 hours, interfering with the creatinine clearance calculation. In the fasting group showed less water intake and lower urinary creatinine. There was great variability for the deproteinized whey and readings performed in the two devices were similar. There was a strong correlation between PT 24 hours and rP/C and the equation was generated: PT 24 hours = (8.6113 x rP/C) + 1.0869. CONCLUSION: Was standardized: 24-hour urine collection without fasting. The deproteinization showed no benefit. The measurements were performed with spectrophotometer reliability. It generated a practical formula for estimating PT 24 hours through rP/C.

Modelos experimentais de obesidade: análise crítica do perfil metabólico e da aplicabilidade / Experimental models of obesity: critical analysis of the metabolic profile and of aplicability

ntrodução: a prevalência da obesidade e de outras doenças relacionadas está aumentando em todo o mundo de forma preocupante. Caracterizada pelo aumento do peso corporal ou do acúmulo excessivo de gordura corporal, a obesidade tem sido associada ao aumento da mortalidade decorrente de maior incidência de hipertensão, diabetes e vários tipos de câncer. Os modelos animais fornecem dados fundamentais para a compreensão dos parâmetros básicos que regulam os componentes do nosso balanço energético. Objetivo: esta revisão selecionou artigos que utilizaram modelos animais (ratos e camundongos) de obesidade focando nas principais alterações metabólicas causadas pela obesidade com o objetivo de apresentar os principais modelos utilizados nos últimos 5 anos. Material e Métodos:Foram realizadas duas buscas na base de dados PubMed utilizando as expressões: "obesity" AND "metabolism" AND "animal model" AND "mice" e "obesity" AND "metabolism" AND "animal model" AND "rat", sendo selecionados os estudos considerados mais relevantes a partir dos critérios: descrição detalhada do modelo experimental e análise dos parâmetros metabólicos de interesse: peso, perfil lipídico e perfil glicêmico. Outras referências foram utilizadas para elucidar melhor os modelos encontrados e também aqueles que não foram citados, mas, que possuem importância no entendimento da evolução dos modelos animais de obesidade. Resultados: A espécie mais utilizada foi o camundongo, o sexo predominante foi o masculino, a faixa etária dos roedores variou de neonatos até 44 semanas e o período de acompanhamento chegou até 53 semanas. A obesidade foi confirmada pelo aumento significativo do peso e na maioria dos estudos foram encontradas alterações no metabolismo lipídico e glicêmico. Encontramos cinco grupos de mecanismos de indução da obesidade porém a maioria dos estudos utilizou dietas hiperlipídicas, modelo que mais se assemelha às alterações metabólicas encontradas em humanos. Conclusão: Investigar as causas e efeitos da obesidade induzida em modelos experimentais pode fornecer uma melhor compreensão da fisiopatologia da obesidade, e proporcionar novas opções de prevenção e tratamento.

Introduction: The prevalence of obesity and other related diseases is increasing worldwide worrisome. Characterized by increased body weight or excessive accumulation of body fat, obesity has been associated with increased mortality due to higher incidence of hypertension, diabetes and many types of cancer. Animal models provide key data for understanding the basic parameters that regulate the components of our energy balance. Objective: This review selected articles that used animal models (mice and mice) for obesity focusing on the main metabolic changes caused by obesity in order to present the main models used in the last 5 years. Material and Methods: Two searches were performed in the PubMed database using the expressions "obesity" AND "metabolism" AND "animal model" AND "mice" and "obesity" AND "metabolism" AND "animal model" AND "rat". We selected the papers considered more relevant from the criteria: detailed description of the experimental model and analysis of the metabolic parameters of interest: weight, lipid profile and glycemic profile. Other references were used to better elucidate the models found and also those that were not mentioned, but we considered important for the understanding of the evolution of animal models of obesity. Results: Mouse was the most used species, the predominant gender was male, the rodent age ranged from neonates up to 44 weeks and the follow-up period reached up to 53 weeks. Obesity was confirmed by a significant weight gain and most studies showed alterations in lipid and glycemic metabolism. We found five groups of induction of obesity, but the majority of studies applied hiperlipidic diets, model which better mimics metabolic chances found in humans. Conclusion: Investigating the causes and effects of induced obesity in experimental models may provide a better understanding of the pathophysiology of obesity, creating new options for prevention and treatment.

Padronização da avaliação da função renal de ratos (Rattus norvegicus) Wistar do biotério da Universidade Federal de Juiz de Fora / Standardization of renal function evaluation in Wistar rats (Rattus norvegicus) from the Federal University of Juiz de Fora's colony

Introdução: Há grande interesse na utilização de modelos animais na pesquisa da fisiopatologia renal, que requer padronização dos parâmetros analisados. Objetivo: Padronizar avaliação da função renal de ratos da colônia do biotério do Centro de Biologia da Reprodução da Universidade Federal de Juiz de Fora. Métodos: Foram utilizados 30 ratos Wistar e realizadas dosagens de creatinina (sérica e urinária), ureia sérica e proteinúria. Foram avaliados: o intervalo de coleta de urina nas gaiolas metabólicas (24 horas ou 12 horas); a necessidade de jejum de 12 horas; a necessidade de desproteinização das amostras de urina e soro para dosagens de creatinina; necessidade de desproteinização do soro de animais com injúria renal aguda (IRA) para leitura em espectrofotômetro e ELISA, além da comparação da proteinúria de 24 horas (PT 24 horas) com a relação proteína/creatinina (rP/C). Os resultados foram comparados pelos teste t de Student, correlação de Pearson, gráfico de Bland-Altman para concordância e modelo de regressão linear para estimar a PT 24 horas a partir da rP/C. Resultados: A diurese de 24 horas foi maior do que a de 12 horas, interferindo na depuração da creatinina. No grupo em jejum, houve menor ingestão hídrica e menor creatinina urinária. Houve grande variabilidade para o soro desproteinizado e as leituras realizadas nos dois equipamentos foram semelhantes. Houve forte correlação entre PT 24 horas e rP/C e foi gerada a equação: PT 24 horas = (8,6113 x rP/C) + 1,0869. Conclusão: Foi padronizada: coleta de urina em 24 horas sem jejum. A desproteinização não mostrou benefício. As dosagens foram realizadas com confiabilidade em espectrofotômetro. Foi ...

Introduction: There is great interest in the use of animal models in the study of renal pathophysiology requires standardization of parameters. Objective: Standardize assessment of renal function in rats from in the Center for Reproductive Biology of Federal University of Juiz de Fora's colony. Methods: Thirty Wistar rats were used and performed measurements of creatinine (serum and urine), serum urea and proteinuria. Were evaluated: the urine collection interval in metabolic cages (24 hours or 12 hours), the need for 12-hour fast, the need of urine and serum deproteinization for creatinine measurement, need of serum deproteinization in animals with acute kidney injury to a spectrophotometer and ELISA, and the comparison of 24-hour proteinuria (PT 24 hours) with the protein/creatinine ratio (rP/C). Means were compared by the Student's t test, Pearson correlation, Bland-Altman plot for agreement and linear regression model to estimate PT 24 hours from rP/C. Results: The 24 hours urine output was greater than 12 hours, interfering with the creatinine clearance calculation. In the fasting group showed less water intake and lower urinary creatinine. There was great variability for the deproteinized whey and readings performed in the two devices were similar. There was a strong correlation between PT 24 hours and rP/C and the equation was generated: PT 24 hours = (8.6113 x rP/C) + 1.0869. Conclusion: Was standardized: 24-hour urine collection without fasting. The deproteinization showed no benefit. The measurements were performed with spectrophotometer reliability. It generated a practical formula for estimating PT 24 hours through rP/C. .