RESUMEN
Copaifera lucens n-butanolic fraction (BF) was used as a source of galloylquinic acids, and aerobically incubated with Aspergillus alliaceus ATCC10060, Aspergillus brasiliensis ATCC 16404, and Cunninghamella elegans ATCC 10028b cultures for 60 and 120 h. Out of the three studied filamentous fungi, A. alliaceus ATCC10060 was able to degrade galloylquinic acids into one major metabolite, 3-O-methylgallic acid (M1). The product was identified by 1H-NMR, UPLC-MS/MS and its potential effect on calcium oxalate monohydrate (COM) crystal binding to Madin-Darby canine kidney cells type I surface was studied. Renal cells pretreatment with BF and M1 for 3 h significantly decreased calcium oxalate monohydrate crystal-adherence at 50 µg/mL and 5 µM, respectively. Both M1 and BF significantly reduced surface expression of COM-binding proteins annexin A1 and heat shock protein 90, respectively as evidenced by Western blot analysis of membrane, cytosolic, and whole cell lysate fractions. The compounds also showed antioxidant activities in DPPH assay.
RESUMEN
The plant metabolite 3,4,5-tri-O-galloylquinic acid methyl ester (TGAME, compound 6) was synthesized, and its potential effect on calcium oxalate monohydrate (COM) crystal binding to the surface of Madin-Darby canine kidney cells type I (MDCKI) and crystal growth in a Drosophila melanogaster Malpighian tubule (MT) model were investigated. Membrane, cytosolic, and total annexin A1 (AxA1), α-enolase, and heat shock protein 90 (HSP90) amounts were examined by Western blot analysis after subcellular fractionation, then confirmed by immunofluorescence staining of cultured cells. Pretreatment of MDCKI cells with TGAME for up to 6 h significantly diminished COM crystal binding in a concentration-dependent manner. TGAME significantly inhibited AxA1 surface expression by immunofluorescence microscopy, whereas intracellular AxA1 increased. Western blot analysis confirmed AxA1 expression changes in the membrane and cytosolic fractions of compound-treated cells, whereas whole cell AxA1 remained unchanged. TGAME also significantly decreased the size, number, and growth of calcium oxalate (CaOx) crystals induced in a Drosophila melanogaster MT model and possessed a potent antioxidant activity in a DPPH assay.
Asunto(s)
Anexina A1/efectos de los fármacos , Oxalato de Calcio/química , Adhesión Celular/efectos de los fármacos , Ácido Gálico/análogos & derivados , Ácido Quínico/análogos & derivados , Animales , Anexina A1/metabolismo , Antioxidantes , Línea Celular , Cristalización , Perros , Drosophila melanogaster , Ácido Gálico/síntesis química , Ácido Gálico/química , Ácido Gálico/farmacología , Células de Riñón Canino Madin Darby/efectos de los fármacos , Células de Riñón Canino Madin Darby/metabolismo , Túbulos de Malpighi/química , Ácido Quínico/síntesis química , Ácido Quínico/química , Ácido Quínico/farmacología , Fracciones Subcelulares/química , Fracciones Subcelulares/metabolismoRESUMEN
Antecedentes: La guía Kidney Disease Improving Global Outcomes (KDIGO) recomienda el uso de un filtrado glomerular estimado (IFGe) basado en cistatina-C para confirmar un IFGe basado en creatinina entre 45 y 59 mL ∙ min-1 ∙ (1,73 m2)-1. Estudios anteriores han demostrado que comorbilidades tales como trasplantes de órganos sólidos influyen fuertemente en la relación entre el índice de filtrado glomerular (IFG) medido, creatinina y cistatina-C. Nuestro objetivo fue evaluar el desempeño de ecuaciones de IFGe basadas en cistatina-C en comparación con IFG medido e IFGe basada en creatinina en diferentes situaciones clínicas. Métodos: Comparamos el rendimiento de la ecuación CKD-EPI 2009, ecuación de IFGe basada en creatinina (IFGeCr), y las nuevas ecuaciones CKD-EPI 2012 basadas en cistatina-C (IFGeCys y IFGeCr-Cys), con el IFG medido (depuración renal de iotalamato) en poblaciones de pacientes totalmente definidas. Los pacientes (n=1.652) fueron clasificados como receptores de trasplantes (de riñón, n=568 u otro órgano, n=319), pacientes con enfermedad renal crónica (ERC) conocida (n=618), o potenciales donantes de riñón (n=147). Resultados: IFGeCr-Cys mostró el desempeño más consistente a través de diferentes poblaciones clínicas. Entre los potenciales donantes de riñón sin ERC [estadio 2 o superior; IFGe> 60 mL ∙ min-1 ∙ (1,73 m2)-1], IFGeCys y IFGeCr‑Cys demostraron significativamente menor sesgo que IFGeCr; sin embargo, las 3 ecuaciones subestimaron substancialmente IFG cuando IFGe fue <60 mL ∙ min-1 ∙ (1,73 m2)-1. Entre los receptores de trasplante con ERC estadio 3B o mayor [IFGe <45 mL ∙ min-1 ∙ (1,73 m2)-1], IFGeCys mostró un sesgo significativamente mayor que IFGeCr. No se observaron diferencias claras en el sesgo de IFGe entre las ecuaciones dentro del grupo de pacientes con ERC conocida independientemente del rango de IFGe o en cualquier grupo de pacientes con una IFG entre 45 y 59 mL ∙ min-1 ∙ (1,73 m2)-1. Conclusiones: El desempeño de las ecuaciones de IFGe depende de las características del paciente las cuales son evidentes en su presentación. Entre las 3 ecuaciones CKD-EPI, IFGeCr-Cys tuvo un desempeño más coherente en las poblaciones de pacientes estudiados.
Background: The Kidney Disease Improving Global Outcomes (KDIGO) guideline recommends use of a cystatin C-based estimated glomerular filtration rate (eGFR) to confirm creatinine-based eGFR between 45 and 59 mL ∙ min-1 ∙ (1.73 m2)-1. Prior studies have demonstrated that comorbidities such as solid-organ transplant strongly influence the relationship between measured GFR, creatinine, and cystatin C. Our objective was to evaluate the performance of cystatin C-based eGFR equations compared with creatinine-based eGFR and measured GFR across different clinical presentations. Methods: We compared the performance of the CKD-EPI 2009 creatinine-based estimated GFR equation (eGFRCr) and the newer CKD-EPI 2012 cystatin C-based equations (eGFRCys and eGFRCr-Cys) with measured GFR (iothalamate renal clearance) across defined patient populations. Patients (n = 1652) were categorized as transplant recipients (n = 568 kidney; n = 319 other organ), known chronic kidney disease (CKD) patients (n = 618), or potential kidney donors (n = 147). Results: eGFRCr-Cys showed the most consistent performance across different clinical populations. Among potential kidney donors without CKD [stage 2 or higher; eGFR >60 mL ∙ min-1 ∙ (1.73 m2)-1], eGFRCys and eGFRCr-Cys demonstrated significantly less bias than eGFRCr; however, all 3 equations substantially underestimated GFR when eGFR was <60 mL ∙ min-1 ∙ (1.73 m2)-1. Among transplant recipients with CKD stage 3B or greater [eGFR <45 mL ∙ min-1 ∙ (1.73 m2)-1], eGFRCys was significantly more biased than eGFRCr. No clear differences in eGFR bias between equations were observed among known CKD patients regardless of eGFR range or in any patient group with a GFR between 45 and 59 mL ∙ min-1 ∙ (1.73 m2)-1. Conclusions: The performance of eGFR equations depends on patient characteristics that are readily apparent on presentation. Among the 3 CKD-EPI equations, eGFRCr-Cys performed most consistently across the studied patient populations.
Asunto(s)
Humanos , Creatina , Cistatina C , Tasa de Filtración Glomerular , Creatinina , Insuficiencia Renal CrónicaRESUMEN
ANTECEDENTES: A excreção urinária de albumina indica lesão nos rins e é reconhecida como fator de risco para a progressão das doenças renal e cardiovascular. A dosagem da albumina urinária chama a atenção sobre a necessidade clínica de relatos de resultados precisos e claramente descritos. O National Kidney Disease Education Program e a Federação Internacional de Química Clínica e Medicina Laboratorial (IFCC) reuniram-se para avaliar o estado atual das questões pré-analíticas, analíticas e pós-analíticas que afetam as dosagens da albumina na urina e para identificar as áreas que necessitam de melhorias. CONTEÚDO: A química da albumina na urina não é completamente compreendida. Diretrizes atuais recomendam a utilização da relação albumina/creatinina (RAC) como substituta para a coleta de amostras cronometradas de urina, frequentemente inadequadas. Os resultados da RAC são afetados pela preparação do paciente, pela hora do dia da coleta das amostras e não é padronizada. Foram relatadas consideráveis diferenças intermétodos para a dosagem tanto de albumina quanto de creatinina, mas a verdade é desconhecida, porque não existem procedimentos de referência para a dosagem de albumina e não há materiais de referência para qualquer um desses analitos na urina. Os intervalos de referência recomendados para a RAC não consideram as grandes diferenças intergrupos na excreção da creatinina (por exemplo, relacionadas com diferenças em idade, sexo e etnia), nem o aumento contínuo no risco relacionado com a excreção de albumina. DISCUSSÃO: Necessidades clínicas foram identificadas para a padronização de (a) métodos de coleta da urina, (b) dosagens de albumina e de creatinina na urina com base em um sistema de referência completo, (c) relatórios dos resultados dos testes e (d) intervalos de referência para a RAC.
BACKGROUND: Urinary excretion of albumin indicates kidney damage and is recognized as a risk factor for progression of kidney disease and cardiovascular disease. The role of urinary albumin measurements has focused attention on the clinical need for accurate and clearly reported results. The National Kidney Disease Education Program and the IFCC convened a conference to assess the current state of preanalytical, analytical, and postanalytical issues affecting urine albumin measurements and to identify areas needing improvement. CONTENT: The chemistry of albumin in urine is incompletely understood. Current guidelines recommend the use of the albumin/creatinine ratio (ACR) as a surrogate for the error-prone collection of timed urine samples. Although ACR results are affected by patient preparation and time of day of sample collection, neither is standardized. Considerable intermethod differences have been reported for both albumin and creatinine measurement, but trueness is unknown because there are no reference measurement procedures for albumin and no reference materials for either analyte in urine. The recommended reference intervals for the ACR do not take into account the large intergroup differences in creatinine excretion (e.g., related to differences in age, sex, and ethnicity) nor the continuous increase in risk related to albumin excretion. DISCUSSION: Clinical needs have been identified for standardization of (a) urine collection methods, (b) urine albumin and creatinine measurements based on a complete reference system, (c) reporting of test results, and (d) reference intervals for the ACR.