Untargeted metabolomics analysis of serum and urine unveils the protective effect of cilastatin on altered metabolic pathways during cisplatin-induced acute kidney injury.

Acute kidney injury (AKI) is one of the most serious complications of cisplatin anticancer therapies. Cilastatin is a highly promising nephroprotective agent to eventually enter clinical use, but its biochemical mechanism is still not fully understood. We have employed an untargeted metabolomics approach based on capillary electrophoresis mass spectrometry (CE-MS) analysis of serum and urine from an in vivo rat model, to explore the metabolic pathways involved in cisplatin-induced AKI and cilastatin nephroprotection. A total of 155 and 76 identified metabolites were found to be significantly altered during cisplatin treatment in urine and serum, respectively. Most of these altered metabolites were either partially or totally recovered by cilastatin and cisplatin co-treatment. The main metabolic pathways disturbed by cisplatin during AKI involved diverse amino acids metabolism and biosynthesis, tricarboxylic acids (TCA) cycle, nicotinate and nicotinamide metabolism, among others. Cilastatin was proved to protect diverse cisplatin-altered pathways involving metabolites related to immunomodulation, inflammation, oxidative stress and amino acid metabolism in proximal tubules. However, cisplatin-altered mitochondrial metabolism (especially, the energy-producing TCA cycle) remained largely unprotected by cilastatin, suggesting an unresolved mitochondrial direct damage. Multivariate analysis allowed effective discrimination of cisplatin-induced AKI and cilastatin renoprotection based on metabolic features. A number of potential serum and urine biomarkers could also be foreseen for cisplatin-induced AKI detection and cilastatin nephroprotection.

Tubular biomarkers and histology in lupus nephritis.

INTRODUCTION: The relevance of tubulo-interstitial involvement for kidney prognosis has recently been emphasized, but validated biomarkers for predicting histology are still lacking. The aim of our study was to evaluate different serum and urinary markers of tubular damage in patients with lupus nephritis (LN) and to correlate them with kidney histopathology. METHODS: A single-center retrospective study was conducted from January 2016 to December 2021. Serum and urine samples were collected on the same day of kidney biopsy and correlated with histologic data from a cohort of 15 LN patients. We analyzed the following urinary markers, adjusted for urine creatinine: beta 2-microglobulin, alpha 1-microglobulin, NGAL, uKIM-1, MCP-1, uDKK-3, and uUMOD. The serum markers sKIM-1 and sUMOD were also analyzed. RESULTS: A positive and strong correlation was observed between the degree of interstitial fibrosis (rho = 0.785, p = .001) and tubular atrophy (rho = 0.781, p = .001) and the levels of uDKK3. uUMOD also showed an inverse and moderate correlation with interstitial fibrosis (rho = -0.562, p = .037) and tubular atrophy (rho = -0.694, p = .006). Patients with >10% cortical interstitial inflammation had higher levels of uKIM-1 [4.9 (3.9, 5.5) vs. 0.8 (0.6, 1.5) mcg/mg, p = .001], MCP-1 [3.8 (2. 3, 4.2) vs. 0.7 (0.3, 1.2) mcg/mg, p = .001], sKIM-1 [9.2 (5.9, 32.7) vs. 1.4 (0, 3.5) pg/mL, p = .001], and lower sUMOD [8.7 (0, 39.7) vs. 46.1 (35.7, 53) ng/mL, p = .028]. CONCLUSION: The use of specific urinary and serum biomarkers of tubular dysfunction or injury may help to predict certain histologic parameters in LN patients.

Tubular biomarkers in proteinuric kidney disease: histology correlation and kidney prognosis of tubular biomarkers.

Background: Proteinuria is not only a biomarker of chronic kidney disease (CKD) but also a driver of CKD progression. The aim of this study was to evaluate serum and urinary tubular biomarkers in patients with biopsied proteinuric kidney disease and to correlate them with histology and kidney outcomes. Methods: A single-center retrospective study was conducted on a cohort of 156 patients from January 2016 to December 2021. The following urinary and serum biomarkers were analyzed on the day of kidney biopsy: beta 2 microglobulin (ß2-mcg), alpha 1 microglobulin (α1-mcg), neutrophil gelatinase-associated lipocalin (NGAL), urinary kidney injury molecule-1 (uKIM-1), monocyte chemoattractant protein-1 (MCP-1), urinary Dickkopf-3 (uDKK3), uromodulin (urinary uUMOD), serum kidney injury molecule-1 (sKIM-1) and serum uromodulin (sUMOD). A composite outcome of kidney progression or death was recorded during a median follow-up period of 26 months. Results: Multivariate regression analysis identified sUMOD (ß-0.357, P < .001) and uDKK3 (ß 0.483, P < .001) as independent predictors of interstitial fibrosis, adjusted for age, estimated glomerular filtration rate (eGFR) and log proteinuria. Elevated levels of MCP-1 [odds ratio 15.61, 95% confidence interval (CI) 3.52-69.20] were associated with a higher risk of cortical interstitial inflammation >10% adjusted for eGFR, log proteinuria and microhematuria. Upper tertiles of uDKK3 were associated with greater eGFR decline during follow-up. Although not a predictor of the composite outcome, doubling of uDKK3 was a predictor of kidney events (hazard ratio 2.26, 95% CI 1.04-4.94) after adjustment for interstitial fibrosis, eGFR and proteinuria. Conclusions: Tubular markers may have prognostic value in proteinuric kidney disease, correlating with specific histologic parameters and identifying cases at higher risk of CKD progression.

Cilastatin as a Potential Anti-Inflammatory and Neuroprotective Treatment in the Management of Glaucoma.

Glaucoma is a neurodegenerative disease that causes blindness. In this study, we aimed to evaluate the protective role of cilastatin (CIL), generally used in the treatment of nephropathologies associated with inflammation, in an experimental mouse model based on unilateral (left) laser-induced ocular hypertension (OHT). Male Swiss mice were administered CIL daily (300 mg/kg, i.p.) two days before OHT surgery until sacrifice 3 or 7 days later. Intraocular Pressure (IOP), as well as retinal ganglion cell (RGC) survival, was registered, and the inflammatory responses of macroglial and microglial cells were studied via immunohistochemical techniques. Results from OHT eyes were compared to normotensive contralateral (CONTRA) and naïve control eyes considering nine retinal areas and all retinal layers. OHT successfully increased IOP values in OHT eyes but not in CONTRA eyes; CIL did not affect IOP values. Surgery induced a higher loss of RGCs in OHT eyes than in CONTRA eyes, while CIL attenuated this loss. Similarly, surgery increased macroglial and microglial activation in OHT eyes and to a lesser extent in CONTRA eyes; CIL prevented both macroglial and microglial activation in OHT and CONTRA eyes. Therefore, CIL arises as a potential effective strategy to reduce OHT-associated damage in the retina of experimental mice.

Novel Experimental Mouse Model to Study Malaria-Associated Acute Kidney Injury.

The impact of malaria-associated acute kidney injury (MAKI), one of the strongest predictors of death in children with severe malaria (SM), has been largely underestimated and research in this area has been neglected. Consequently, a standard experimental mouse model to research this pathology is still lacking. The purpose of this study was to develop an in vivo model that resembles the pathology in MAKI patients. In this study, unilateral nephrectomies were performed on wild-type mice prior to infection with Plasmodium berghei NK65. The removal of one kidney has shown to be an effective approach to replicating the most common findings in humans with MAKI. Infection of nephrectomized mice, compared to their non-nephrectomized counterparts, resulted in the development of kidney injury, evident by histopathological analysis and elevated levels of acute kidney injury (AKI) biomarkers, including urinary neutrophil gelatinase-associated lipocalin, serum Cystatin C, and blood urea nitrogen. Establishment of this in vivo model of MAKI is critical to the scientific community, as it can be used to elucidate the molecular pathways implicated in MAKI, delineate the development of the disease, identify biomarkers for early diagnosis and prognosis, and test potential adjunctive therapies.

Trends in hospitalization for urinary tract infection in the paediatric age group in the 2000-2015 period in Spain.

OBJECTIVE: To analyse the trends in hospital admissions related to urinary tract infection among children aged 0-14 years in Spain in the 2000-2015 period. METHODS: We conducted a retrospective observational study using the minimum basic hospital discharge dataset system of Spain, which applies the International Classification of Diseases, version 9 (ICD-9) coding system. We included every hospitalisation due to cystitis, pyelonephritis and unspecified UTI among children aged less than 15 years. We collected data on patient sex and age, type of discharge, main diagnosis, comorbidities, length of stay and overall cost. We calculated crude hospitalization rates per 1000 inhabitants aged less than 15 years and performed a joinpoint regression analysis to identify temporal trends. RESULTS: In the 2000-2015 period, there were 124 696 hospitalizations in children under 15 years. Of these patients, 72.97% were aged 0-1 year and 60.12% had a diagnosis of unspecified UTI, 39.27% of pyelonephritis, and 0.52% of cystitis. The crude rate of hospitalization due to UTI ranged from 1.24 in year 2000 to 0.98 in 2015. The rate of hospitalization was higher in female versus male patients. The joinpoint analysis found a decreasing trend in the rate of hospitalization due to UTI, with an average annual percent change (AAPC) of -1.5% (95% confidence interval [CI], -2.4 a -0.6). The largest decreases occurred in female patients (AAPC, -1.8; 95% CI, -2.5 a -1.0) and children aged 7-10 years (AAPC - 5.9; 95% CI, -6.7 a -5.2). CONCLUSIONS: The rate of hospitalization related to UTI in Spain in patients aged up to 14 years decreased during the 2000-2015 period. The highest hospitalization rates occurred in female patients and in the 0-to-1 year age group.

FKBP51, AmotL2 and IQGAP1 Involvement in Cilastatin Prevention of Cisplatin-Induced Tubular Nephrotoxicity in Rats.

The immunophilin FKBP51, the angiomotin AmotL2, and the scaffoldin IQGAP1 are overexpressed in many types of cancer, with the highest increase in leucocytes from patients undergoing oxaliplatin chemotherapy. Inflammation is involved in the pathogenesis of nephrotoxicity induced by platinum analogs. Cilastatin prevents renal damage caused by cisplatin. This functional and confocal microscopy study shows the renal focal-segmental expression of TNFα after cisplatin administration in rats, predominantly of tubular localization and mostly prevented by co-administration of cilastatin. FKBP51, AmotL2 and IQGAP1 protein expression increases slightly with cilastatin administration and to a much higher extent with cisplatin, in a cellular- and subcellular-specific manner. Kidney tubule cells expressing FKBP51 show either very low or no expression of TNFα, while cells expressing TNFα have low levels of FKBP51. AmotL2 and TNFα seem to colocalize and their expression is increased in tubular cells. IQGAP1 fluorescence increases with cilastatin, cisplatin and joint cilastatin-cisplatin treatment, and does not correlate with TNFα expression or localization. These data suggest a role for FKBP51, AmotL2 and IQGAP1 in cisplatin toxicity in kidney tubules and in the protective effect of cilastatin through inhibition of dehydropeptidase-I.

Role of Glucocorticoid Signaling and HDAC4 Activation in Diaphragm and Gastrocnemius Proteolytic Activity in Septic Rats.

Sepsis increases glucocorticoid and decreases IGF-1, leading to skeletal muscle wasting and cachexia. Muscle atrophy mainly takes place in locomotor muscles rather than in respiratory ones. Our study aimed to elucidate the mechanism responsible for this difference in muscle proteolysis, focusing on local inflammation and IGF-1 as well as on their glucocorticoid response and HDAC4-myogenin activation. Sepsis was induced in adult male rats by lipopolysaccharide (LPS) injection (10 mg/kg), and 24 h afterwards, rats were euthanized. LPS increased TNFα and IL-10 expression in both muscles studied, the diaphragm and gastrocnemius, whereas IL-6 and SOCS3 mRNA increased only in diaphragm. In comparison with gastrocnemius, diaphragm showed a lower increase in proteolytic marker expression (atrogin-1 and LC3b) and in LC3b protein lipidation after LPS administration. LPS increased the expression of glucocorticoid induced factors, KLF15 and REDD1, and decreased that of IGF-1 in gastrocnemius but not in the diaphragm. In addition, an increase in HDAC4 and myogenin expression was induced by LPS in gastrocnemius, but not in the diaphragm. In conclusion, the lower activation of both glucocorticoid signaling and HDAC4-myogenin pathways by sepsis can be one of the causes of lower sepsis-induced proteolysis in the diaphragm compared to gastrocnemius.

Ejercicios para mejorar la efectividad del Tokui-Waza en el Tachi Waza / Exercícios para melhorar a eficácia do Tokui-Waza em Tachi Waza / Exercises to improve the effectiveness of Tokui-Waza in Tachi Waza

RESUMEN En el judo actual, el dominio del Tokui-Waza o técnica favorita es imprescindible para vencer al contrario y alzarse con la victoria; no obstante, ello requiere de una adecuada preparación técnico táctica y su sistematización en el proceso de entrenamiento. A partir de la existencia de una serie de insuficiencias relacionadas con el proceder didáctico de los entrenadores para la enseñanza-aprendizaje de las acciones técnico tácticas del Tokui Waza y en aras de posibilitarle a las atletas, una adecuada formación en el proceso de preparación técnica, fundamentado en nuevas y mejores formas de afrontar y perfeccionar este proceso, la presente investigación tuvo como objetivo implementar un conjunto de ejercicios para el mejoramiento de la efectividad del Tokui Waza en el Tachi Waza del equipo femenino de judo, categoría 15-16 años de la Escuela Integral Deportiva de Pinar del Río, en correspondencia con las diferentes manifestaciones dentro del combate. Para ello, se trabajó con una muestra de 12 atletas integrantes de este equipo. Se emplearon los métodos y técnicas de investigación científica como: la observación, entrevista estandarizada e individual y el análisis de documentos. Como resultado se aporta un conjunto de ejercicios estructurados, que incluyen situaciones simplificadas y complejas que actúan didácticamente sobre los componentes de la preparación técnico-táctica además que constituyen una herramienta de utilidad para los entrenadores en el perfeccionamiento de su labor en la preparación deportiva de los atletas.

RESUMO No judô actual, o domínio do Tokui-Waza ou técnica favorita é essencial para derrotar o adversário e ganhar a vitória; contudo, isto requer uma preparação técnico-táctica adequada e a sua sistematização no processo de treino. Da existência de uma série de insuficiências relacionadas com o procedimento didáctico dos treinadores para o ensino-aprendizagem das acções técnico-tácticas de Tokui Waza e a fim de permitir aos atletas, um treino adequado no processo de preparação técnica, baseado em novas e melhores formas de abordar e melhorar este processo, o objectivo desta investigação era implementar um conjunto de exercícios para melhorar a eficácia de Tokui Waza no Tachi Waza da equipa feminina de judô, categoria 15-16 anos da Escuela Integral Deportiva de Pinar del Río, em correspondência com as diferentes manifestações no âmbito do combate. Para este efeito, trabalhou se com uma amostra de 12 atletas membros desta equipa. Foram utilizados métodos e técnicas de investigação científica, tais como: observação, entrevista padronizada e individualizada e análise de documentos. Como resultado, é fornecido um conjunto de exercícios estruturados, que incluem situações simplificadas e complexas que actuam didaticamente sobre os componentes da preparação técnico-táctica e que constituem uma ferramenta útil para os treinadores na melhoria do seu trabalho na preparação desportiva dos atletas.

ABSTRACT In the current sport of Judo, the mastery of the Tokui-Waza or favorite technique is essential to defeat the opponent and win the victory, however this requires an adequate technical-tactical preparation and its systematization in the training process. From the existence of a series of inadequacies related to the didactic procedure of the coaches for the teaching-learning of the technical-tactical actions of Tokui Waza, in order to enable the athletes an adequate training in the process of technical preparation, based on new and better ways to address and improve this process, the objective of this research is to implement a set of exercises to improve the effectiveness of Tokui Waza in the Tachi Waza of the women's Judo team, category 15-16 years old, of the Integral Sports School in Pinar del Río, in correspondence with the different manifestations within the combat. For this purpose, a sample of 12 athletes, members of this team, was used. Scientific research methods and techniques were used, such as: observation, standardized and individual interview, as well as document analysis. As a result, a set of structured exercises is provided, which include simplified and complex situations, which act didactically on the components of the technical-tactical preparation, constituting a useful tool for coaches in the improvement of their work in the sports preparation of athletes.

Effect of Cilastatin on Cisplatin-Induced Nephrotoxicity in Patients Undergoing Hyperthermic Intraperitoneal Chemotherapy.

Cisplatin is one of the most widely used chemotherapeutic agents in oncology, although its nephrotoxicity limits application and dosage. We present the results of a clinical study on prophylaxis of cisplatin-induced nephrotoxicity in patients with peritoneal carcinomatosis undergoing cytoreduction and hyperthermic intraperitoneal intraoperative chemotherapy (HIPEC-cisplatin). Prophylaxis was with imipenem/cilastatin. Cilastatin is a selective inhibitor of renal dehydropeptidase I in the proximal renal tubule cells that can reduce the nephrotoxicity of cisplatin. Unfortunately, cilastatin is not currently marketed alone, and can only be administered in combination with imipenem. The study has a retrospective part that serves as a control (n = 99 patients receiving standard surgical prophylaxis) and a prospective part with imipenem/cilastatin prophylaxis corresponding to the study group (n = 85 patients). In both groups, we collected specific data on preoperative risk factors of renal damage, fluid management, hemodynamic control, and urine volume during surgery (including the hyperthermic chemotherapy perfusion), as well as data on hemodynamic and renal function during the first seven days after surgery. The main finding of the study is that cilastatin may exert a nephroprotective effect in patients with peritoneal carcinomatosis undergoing cytoreduction and hyperthermic intraperitoneal cisplatin perfusion. Creatinine values remained lower than in the control group (ANOVA test, p = 0.037). This translates into easier management of these patients in the postoperative period, with significantly shorter intensive care unit (ICU) and hospital stay.

Ejercicios especiales para mejorar la ejecución de técnicas de piernas en judocas escolares / Exercícios especiais para melhorar a execução de técnicas de pernas em judocas escolares / Special exercises to improve the execution of leg techniques in school judo

Resumen Insuficiencias detectadas en la realización de técnicas de piernas de los judocas escolares de Pinar del Río motivaron la realización de esta investigación que tiene como objetivo: elaborar un conjunto de ejercicios especiales que contribuyan al desarrollo del trabajo de los entrenadores con este grupo de técnicas para mejorar la ejecución de los 14 atletas de esta categoría. Al inicio de este estudio, se realizaron observaciones a sesiones de entrenamiento, topes de control y competencias preparatorias para obtener información directa sobre el modo en que se manifiestan estas acciones. También se utilizó una entrevista a los entrenadores para conocer sus conocimientos, criterios y desempeño en el trabajo con estas técnicas y la encuesta para valorar el conocimiento de los atletas acerca de la importancia de la preparación técnica en estas acciones. Además, se practicó una medición mediante una prueba técnica para conocer el comportamiento de la ejecución de estas en los atletas motivo de investigación, en la cual las variables analizadas fueron: puntuación obtenida, total de acciones y forma de realización (directa, combinación o como contraataque). Este estudio permitió el conocimiento de las deficiencias presentadas por estos atletas en la ejecución de las técnicas de piernas, las cuales influyen de forma negativa en los resultados competitivos ya que estas se realizan con gran frecuencia durante el combate por su vinculación con otras técnicas en la realización de combinaciones y contraataques. Este trabajo es una herramienta valiosa para la labor de los entrenadores, dirigida a la preparación técnica del judoca.

Resumo As insuficiências detectadas no desempenho das técnicas de perna das judocas escolares de Pinar del Río motivaram a realização desta investigação que tem como objetivo: elaborar um conjunto de exercícios especiais que contribuam para o desenvolvimento do trabalho dos treinadores com este grupo de técnicas para melhorar o desempenho dos 14 atletas desta categoria. No início deste estudo, foram feitas observações a sessões de formação, paragens de controlo e concursos preparatórios para obter informações diretas sobre a forma como estas ações se manifestam. Uma entrevista com os treinadores foi também utilizada para descobrir os seus conhecimentos, critérios e desempenho no trabalho com estas técnicas e o inquérito foi utilizado para avaliar os conhecimentos dos atletas sobre a importância da preparação técnica nestas ações. Além disso, foi feita uma medição através de um teste técnico para conhecer o comportamento da execução destes nos atletas sob investigação, no qual as variáveis analisadas foram: pontuação obtida, total de ações e forma de execução (direta, combinada ou como contra-ataque). Este estudo permitiu o conhecimento das deficiências apresentadas por estes atletas na execução das técnicas de pernas, que influenciam de forma negativa nos resultados competitivos, uma vez que estes são feitos com grande frequência durante o combate pela sua implicação com outras técnicas na realização de combinações e contra-ataques. Este trabalho é um instrumento valioso para o trabalho dos treinadores, destinado à preparação técnica do judoca.

Abstract Insufficiencies detected in the performance of leg techniques of the school judokas of Pinar del Río motivated the realization of this research, which aims to develop a set of special exercises that contribute to the development of the work of coaches with this group of techniques to improve the performance of the 14 athletes in this category. At the beginning of this study, observations were made of training sessions, control stops and preparatory competitions to obtain direct information on how these actions are manifested. An interview with the coaches was also used to find out their knowledge, criteria, and performance at work with these techniques, and the survey was used to assess the athletes' knowledge about the importance of technical preparation in these actions. In addition, a measurement was carried out by means of a technical test, to know the behavior of the execution of these in the athletes reason for research, in which the variables analyzed were: score obtained, total of actions and form of performance (direct, combination or as a counterattack). This study found the knowledge of the deficiencies presented by these athletes, in the execution of leg techniques, which negatively influence competitive results since these are performed with great frequency during combat due to their link with other techniques in performing combinations and counterattacks. This work is a valuable tool for the work of the coaches aimed at the technical preparation of the judoka.

Urinary excretion of parathyroid hormone-related protein correlates with renal function in control rats and rats with cisplatin nephrotoxicity.

Parathyroid hormone-related protein (PTHrP) and its receptor are abundantly expressed throughout the renal parenchyma, where PTHrP exerts a modulatory action on renal function. PTHrP upregulation is a common event associated with the mechanism of renal injury and repair. However, no study has yet explored the putative excretion of PTHrP in urine, including its potential relationship with renal function. In the present study, we tested this hypothesis by studying the well-known rat model of acute renal injury induced by the chemotherapeutic agent cisplatin. Using Western blot analysis, we could detect a single protein band, corresponding to intact PTHrP, in the urine of both control and cisplatin-injected rats, whose levels were significantly higher in the latter group. PTHrP was detected in rat urine by dot blot, and its quantification with two specific ELISA kits showed that, compared with control rats, those treated with cisplatin displayed a significant increase in urinary PTHrP (expressed as the PTHrP-to-creatinine ratio or 24-h excretion). In addition, a positive correlation between urinary PTHrP excretion and serum creatinine was found in these animals. In conclusion, our data demonstrate that PTHrP is excreted in rat urine and that this excretion is higher with the decrease of renal function. This suggests that urinary PTHrP levels might be a renal function marker.

La interconexión de las cargas en la planificación del entrenamiento, en el deporte de boxeo / The interconnection of the loads in the planning of the training, in the sport of boxing

Resumen El presente trabajo se enmarca en el contexto de las alternativas relacionadas con la planificación del entrenamiento deportivo, a partir de los modelos ATR (acumulación- transformación-realización). En correspondencia con las concepciones que hoy se utilizan en materia de planificación del entrenamiento, se evidencia la importancia de la interconexión de los contenidos en la preparación de los boxeadores, a partir de la conjugación de cargas cuyas rutas metabólicas son fundamentalmente la fosfagénica, anaerobia aláctica y la glicolitica, anaerobia láctica. El objetivo de la investigación radica en valorar el estado de la planificación del entrenamiento, en función de las tendencias más actuales, relacionadas con la interconexión de las cargas y el modelo ATR en la preparación de los boxeadores de la Academia Provincial de Pinar del Río. Para el estudio, se utilizó una muestra conformada por 13 atletas y siete entrenadores de la Academia Provincial de boxeo de Pinar del Río. Se emplearon diferentes técnicas y métodos como la entrevista a entrenadores, la observación y la medición. No obstante, los documentos y orientaciones de la Comisión Nacional de boxeo aún no cubren todas las expectativas de los colectivos pedagógicos de entrenadores de la academia, lo que deja un margen a la interpretación, por parte de las Comisiones Técnicas de los territorios en cuanto a las alternativas, a la planificación del entrenamiento y la definición de las direcciones e interconexión de estas. Resulta necesario encontrar un mecanismo que, en el plano didáctico, oriente mejor al entrenador en este sentido.

Abstract The present work is framed in the context of the alternatives related to the planning of the sport training, from the ATR models (accumulation-transformation-realization). In correspondence with the conceptions used today in training planning, the importance of the interconnection of the contents in the preparation of boxers is evident, from the conjugation of loads whose metabolic routes are fundamentally phosphagenic, alactic anaerobic and lactic anaerobic glycolithic. The objective of the research is to assess the situation of the training planning, based on the most current trends related to the interconnection of the loads and the ATR model in the preparation of the boxers of the Provincial Academy of Pinar del Río. For the study it was selected a sample, consisting on 13 athletes and 7 coaches from the Provincial Boxing Academy of Pinar del Río. Different methods and techniques such as the interview, the observation and the measurement were used. This study was carried outin correspondence with the most advanced conceptions regarding training planning. It reveals the importance of the interconnection of the contents in the preparation of the boxers from the combination of lactic anaerobic loads and lactic ones. However, the documents and guidelines of the National Boxing Commission still do not meet all the expectations of the pedagogical groups of trainers of the academy, which leaves a margin for interpretation by the Technical Commissions of the territories as regards alternatives, the planning of training and the definition of the directions and interconnection of these. It is necessary to find a mechanism that, at the didactic level, better orients the trainer in this sense.

Lipid imaging for visualizing cilastatin amelioration of cisplatin-induced nephrotoxicity.

Nephrotoxicity is a major limitation to cisplatin antitumor therapies. Cilastatin, an inhibitor of renal dehydropeptidase-I, was recently proposed as a promising nephroprotector against cisplatin toxicity, preventing apoptotic cell death. In this work, cilastatin nephroprotection was further investigated in a rat model, with a focus on its effect on 76 renal lipids altered by cisplatin, including 13 new cisplatin-altered mitochondrial cardiolipin species. Lipid imaging was performed with MALDI mass spectrometry imaging (MALDI-MSI) in kidney sections from treated rats. Cilastatin was proved to significantly diminish the lipid distribution alterations caused by cisplatin, lipid levels being almost completely recovered to those of control samples. The extent of recovery of cisplatin-altered lipids by cilastatin turned out to be relevant for discriminating direct or secondary lipid alterations driven by cisplatin. Lipid peroxidation induced by cisplatin was also shown to be reduced when cilastatin was administered. Importantly, significant groups separation was achieved during multivariate analysis of cortex and outer-medullary lipids, indicating that damaged kidney can be discerned from the nephroprotected and healthy groups and classified according to lipid distribution. Therefore, we propose MALDI-MSI as a powerful potential tool offering multimolecule detection possibilities to visualize and evaluate nephrotoxicity and nephroprotection based on lipid analysis.

Cisplatin-induced renal inflammation is ameliorated by cilastatin nephroprotection.

BACKGROUND: Cisplatin is a potent chemotherapeutic drug whose nephrotoxic effect is a major complication and a dose-limiting factor for antitumoral therapy. There is much evidence that inflammation contributes to the pathogenesis of cisplatin-induced nephrotoxicity. We found that cilastatin, a renal dehydropeptidase-I inhibitor, has protective effects in vitro and in vivo against cisplatin-induced renal damage by inhibiting apoptosis and oxidation. Here, we investigated the potential use of cilastatin to protect against cisplatin-induced kidney injury and inflammation in rats. METHODS: Male Wistar rats were divided into four groups: control, cilastatin-control, cisplatin and cilastatin-cisplatin. Nephrotoxicity was assessed 5 days after administration of cisplatin based on blood urea nitrogen, creatinine, glomerular filtration rate (GFR), kidney injury molecule (KIM)-1 and renal morphology. Inflammation was measured using the electrophoretic mobility shift assay, immunohistochemical studies and evaluation of inflammatory mediators. RESULTS: Compared with the control rats, cisplatin-administered rats were affected by significant proximal tubule damage, decreased GFR, increased production of inflammatory mediators and elevations in urea, creatinine and tissue KIM-1 levels. Cilastatin prevented these changes in renal function and ameliorated histological damage in cisplatin-administered animals. Cilastatin also reduced pro-inflammatory cytokine levels, activation of nuclear factor-κB and CD68-positive cell concentrations. CONCLUSIONS: Cilastatin reduces cisplatin-induced nephrotoxicity, which is associated with decreased inflammation in vivo. Although the exact role of decreased inflammation in nephroprotection has not been fully elucidated, treatment with cilastatin could be a novel strategy for the prevention of cisplatin-induced acute kidney injury.

MALDI-LTQ-Orbitrap mass spectrometry imaging for lipidomic analysis in kidney under cisplatin chemotherapy.

Imaging techniques for mapping molecular distributions in tissue sections can reveal valuable information on biomolecules involved in relevant biochemical processes. A method has been developed for comprehensive, reproducible and sensitive lipid imaging by matrix-assisted laser/desorption ionization-LTQ-Orbitrap mass spectrometry in kidney sections, showing the benefits of exact mass determination. Matrix deposition parameters for positive and negative lipid ion imaging using different matrices such as 2,5-dihydroxybenzoic acid (DHB), 9-aminoacridine (9-AA) or α-cyano-4-hydroxycinnamic acid (CHCA) have been optimized for the broadest detection and identification of renal lipids. The combination of 9-AA and DHB was found as the most suitable for negative and positive ion mode lipid imaging, respectively. Lipid mapping and related identification strategies and limitations have also been discussed. Production of 100-µm resolution images was proved to be enough for discerning lipid distribution in kidney substructures. Imaging reproducibility was assessed on parallel kidney slices with time. This method has been applied to the lipidomics analysis on kidney sections from rats treated with the antitumor drug cisplatin and compared to healthy control rats. Up to 66 different renal lipids out of 450 extracted ion images (mainly phospholipid species, in addition to sulfatides and cholesterol sulfate) have been found and identified showing a modified distribution pattern due to cisplatin-induced nephrotoxicity. These lipid species reflect either topographic, signaling or structural processes in damaged kidney and could potentially be used for nephrotoxicity assessment or as therapeutic targets. This is, to our knowledge, the first imaging lipidomics study for nephrotoxicity assessment of cisplatin chemotherapy.

Cisplatin-Induced Non-Oliguric Acute Kidney Injury in a Pediatric Experimental Animal Model in Piglets.

OBJECTIVE: To design an experimental pediatric animal model of acute kidney injury induced by cisplatin. METHODS: Prospective comparative observational animal study in two different phases. Acute kidney injury was induced using three different doses of cisplatin (2, 3 and 5 mg/kg). The development of nephrotoxicity was assessed 2 to 4 days after cisplatin administration by estimating biochemical parameters, diuresis and renal morphology. Analytical values and renal morphology were compared between 15 piglets treated with cisplatin 3 mg/kg and 15 control piglets in the second phase of the study. RESULTS: 41 piglets were studied. The dose of 3 mg/kg administered 48 hours before the experience induced a significant increase in serum creatinine and urea without an increase in potassium levels. Piglets treated with cisplatin 3 mg/kg had significantly higher values of creatinine, urea, phosphate and amylase, less diuresis and lower values of potassium, sodium and bicarbonate than control piglets. Histological findings showed evidence of a dose-dependent increase in renal damage. CONCLUSIONS: a dose of 3 mg/kg of cisplatin induces a significant alteration in renal function 48 hours after its administration, so it can be used as a pediatric animal model of non-oliguric acute kidney injury.

Protective Effects of Cilastatin against Vancomycin-Induced Nephrotoxicity.

Vancomycin is a very effective antibiotic for treatment of severe infections. However, its use in clinical practice is limited by nephrotoxicity. Cilastatin is a dehydropeptidase I inhibitor that acts on the brush border membrane of the proximal tubule to prevent accumulation of imipenem and toxicity. The aim of this study was to investigate the potential protective effect of cilastatin on vancomycin-induced apoptosis and toxicity in cultured renal proximal tubular epithelial cells (RPTECs). Porcine RPTECs were cultured in the presence of vancomycin with and without cilastatin. Vancomycin induced dose-dependent apoptosis in cultured RPTECs, with DNA fragmentation, cell detachment, and a significant decrease in mitochondrial activity. Cilastatin prevented apoptotic events and diminished the antiproliferative effect and severe morphological changes induced by vancomycin. Cilastatin also improved the long-term recovery and survival of RPTECs exposed to vancomycin and partially attenuated vancomycin uptake by RPTECs. On the other hand, cilastatin had no effects on vancomycin-induced necrosis or the bactericidal effect of the antibiotic. This study indicates that cilastatin protects against vancomycin-induced proximal tubule apoptosis and increases cell viability, without compromising the antimicrobial effect of vancomycin. The beneficial effect could be attributed, at least in part, to decreased accumulation of vancomycin in RPTECs.

Cilastatin protects against cisplatin-induced nephrotoxicity without compromising its anticancer efficiency in rats.

Cisplatin is an anticancer agent marred by nephrotoxicity; however, limiting this adverse effect may allow the use of higher doses to improve its efficacy. Cilastatin, a small molecule inhibitor of renal dehydropeptidase I, prevents proximal tubular cells from undergoing cisplatin-induced apoptosis in vitro. Here, we explored the in vivo relevance of these findings and the specificity of protection for kidney cells in cisplatin-treated rats. Cisplatin increased serum blood urea nitrogen and creatinine levels, and the fractional excretion of sodium. Cisplatin decreased the glomerular filtration rate, promoted histological renal injury and the expression of many pro-apoptotic proteins in the renal cortex, increased the Bax/Bcl2 ratio, and oxidative stress in kidney tissue and urine. All these features were decreased by cilastatin, which preserved renal function but did not modify the pharmacokinetics of cisplatin area under the curve. The cisplatin-induced death of cervical, colon, breast, and bladder-derived cancer cell lines was not prevented by cilastatin. Thus, cilastatin has the potential to prevent cisplatin nephrotoxicity without compromising its anticancer efficacy.