Efecto sobre el neurodesarrollo y neuroprotección en pez cebra de un extracto polifenólico de huesos de aceituna

Objetivo: Determinar el efecto de un extracto polifenólico de hueso de oliva en el desarrollo del sistema nervioso y frente al daño inducido mediante la neurotoxina ácido kaínico, utilizando como modelo animal el pez cebra. Material y métodos: Se analiza el efecto del extracto a la máxima dosis tolerada (100 mg/ml de polifenoles) sobre la actividad colinèrgica en larvas de pez cebra (72 horas post-fertilización). Se utilizan únicamente huevos fecundados sin anomalías. Se incuban 6 huevos/pocillo en microplaca de 24 pocillos en 2 ml de agua con DMSO (0,1%) a 26 ± 1º C: a) neurodesarrollo: agua (control) y con 100 mg/ml de extracto, como ensayo; b) neuroprotección: agua y ácido kaínico (100 μM) (control) y con 100 mg/ml de extracto (ensayo). Todas las incubaciones por triplicado. A las 72 h se examinan y verifica ausencia de anomalías. Las larvas se homogeneizan y en los sobrenadantes se cuantifica actividad acetilnolinesterasa y concentración proteínas. Resultados: La cantidad de proteína y apreciación morfológica es análoga en todos los ensayos, indicando mismo desarrollo. La acetilcolinesterasa en las larvas de pez, con el extracto polifenólico es del 162,2%(SD 44,2) respecto a controles (100% de actividad) (p < 0,01). Las larvas de pez tratadas con ácido kaínico y extracto polifenólico presentan el 140,1% (SD 22,0) de actividad, respecto a las incubadas únicamente con la neurotoxina (100%) (p < 0,05). Conclusión: Los polifenoles extraídos de los huesos de aceituna producen incremento de actividad colinèrgica durante el neurodesarrollo larvario en el pez cebra y protección frente a la neurotoxina ácido kaínico.

Accuracy of ASGE criteria for the prediction of choledocholithiasis

Background/aims: Few studies have validated the performance of guidelines for the prediction of choledocholithiasis (CL). Our objective was to prospectively assess the accuracy of the American Society for Gastrointestinal Endoscopy (ASGE) guidelines for the identification of CL. Methods: A two-year prospective evaluation of patients with suspected CL was performed. We evaluated the ASGE guidelines and its component variables in predicting CL. Results: A total of 256 patients with suspected CL were analyzed. Of the 208 patients with high-probability criteria for CL, 124 (59.6%) were found to have a stone/sludge at endoscopic retrograde cholangiopancreatography (ERCP). Among 48 patients with intermediate-probability criteria, 21 (43.8%) had a stone/sludge. The performance of ASGE high- and intermediate-probability criteria in our population had an accuracy of 59.0% (85.5% sensitivity, 24.3% specificity) and 41.0% (14.4% sensitivity, 75.6% specificity), respectively. The mean ERCP delay time was 6.1 days in the CL group and 6.4 days in the group without CL, p = 0.638. The presence of a common bile duct (CBD) > 6 mm (OR 2.21; 95% CI, 1.20-4.10), ascending cholangitis (OR 2.37; 95% CI, 1.01-5.55) and a CBD stone visualized on transabdominal US (OR 3.33; 95% CI, 1.48-7.52) were stronger predictors of CL. The occurrence of biliary pancreatitis was a strong protective factor for the presence of a retained CBD stone (OR 0.30; 95% CI, 0.17-0.55). Conclusions: Irrespective of a patient's ASGE probability for CL, the application of current guidelines in our population led to unnecessary performance of ERCPs in nearly half of cases.

Protection by polyphenol extract from olive stones against apoptosis produced by oxidative stress in human neuroblastoma cells

Objective: We evaluated the protective activity of an extract from a by-product such as olive stones, through its ability to inhibit H2O2 induced apoptosis in the SH-SY5Y human neuroblastoma cell line. Material and methods: To such end, 20,000 cells/well were cultivated and differentiation with retinoic acid was initiated. Once the cells were differentiated, apoptosis was induced with and without H2O2 extract. Finally, cDNA extraction was performed, and pro-apoptotic genes Bax and anti-apoptotic genes Bcl-2 were analyzed. Quantification of the gene expression was performed using the GAPDH gene marker. Results: Cell viability with the extract is 97.6% (SD 5.7) with 10 mg/l and 62.8% (SD 1.2) to 50 mg/l, using 10 mg/l for the biomarker assay. The retinoic acid differentiated SH-S cell line (10 µM) shows a clear apoptosis when treated with H2O2 150 µM, with a Bax/Bcl-2 ratio of 3.75 (SD 0.80) in contrast to the differentiated control cells subjected to H2O2 and with extract, which have the same ratio of 1.02 (SD 0.01-0.03). Conclusion: The olive stone extract shows anti-apoptotic activity in the provoked cell death of SH-SY5Y human neuroblastoma cells in their normal state, defending them from oxidative stress which produces a significant increase in the apoptotic gene ratio in contrast to anti-apoptotic genes (Bax/Bcl-2).