Estudio de la actividad de la lipoproteina lipasa en pacientes con mutaciones APOA5 con hiperlipoproteinemia tipo I

Nuestro objetivo fue estudiar la actividad de la lipoproteína lipasa en pacientes con mutaciones en APOA5 con presencia clara de hiperlipoproteinemia tipo I (HPLI). Material y métodos: Se realizó un estudio de actividad LPL en 2 pacientes con mutaciones en APOA5 (1: homocigoto c.758T>C; 2: doble heterocigoto c.758T>C & c.326_327insC) sin variantes patogénicas en LPL, GPIHBP1, LMF1 y APOC2. Para cada muestra y un control sano, se efectuó un ensayo convencional de actividad de LPL postheparina utilizando como activador un plasma preheparina inactivado de sujetos normolipémicos. En un segundo ensayo, para cada paciente y un control sano, se utilizó plasma preheparina inactivado de los dos sujetos con mutación de APOA5. La presencia de HPLI fue analizada mediante ultracentrifugación secuencial y cálculo del cociente de triglicéridos en quilomicrones entre triglicéridos en VLDL. Resultados: En nuestro laboratorio, la actividad de LPL postheparina en sujetos normolipémicos es de 56±23mU/mL (deficitarioC mostró actividad LPL normal (94±13mU/mL), cuando fue ensayada con plasma preheparina control. Sin embargo, su plasma preheparina fue incapaz de activar la LPL postheparina propia (16±20mU/mL) y la del control (34±17mU/mL). En el paciente heterocigoto c.758T>C & c.326_327insC, al contrario que el anterior, mostró una actividad LPL casi nula (4±7) cuando fue ensayada con plasma preheparina control. El plasma preheparina de este paciente tampoco fue capaz de activar la LPL postheparina propia (0 mU/mL) y del sujeto control sano (22±7mU/mL). Conclusiones: Mutaciones de APOA5 son responsables de HLP tipo I. Estas mutaciones muestran valores dispares de actividad in vitro de LPL postheparina. En ambos casos su suero preheparina no es capaz de activar la LPL propia ni la de sujetos sanosUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

In vivo and In vitro cartilage differentiation from embryonic epicardial progenitor cells

The presence of cartilage tissue in embryonic and adult hearts of various vertebrate species is a well-recorded fact. However, while the embryonic neural crest has been historically considered as the main source of cardiac cartilage, recently reported results on the wide connective potential of epicardial lineage cells suggest they could also differentiate into chondrocytes. During heart embryogenesis, the epicardial epithelium forms over the originally bare myocardial surface from epicardial progenitor (proepicardial) cells to then give rise to a large population of mesenchymal Epicardial-Derived Cells (EPDCs) that will crucially contribute to the building, growth, and maturation of the ventricle and atrioventricular cardiac structures. In this work, we describe the formation of cardiac cartilage clusters from proepicardial cells, both in vivo and in vitro. Our findings report, for the first time, cartilage formation from epicardial progenitor cells in the embryonic heart, and strongly support the concept of proepicardial cells as multipotent connective progenitors. These results are relevant to our understanding of cardiac cell complexity and responses to pathologic stimuli.Universidad de Málaga (UMA18-FEDERJA-146). Campus de Excelencia Internacional Andalucía Tech; Ministerio de Educación (FPU18/05219); Ministerio de Ciencias (RTI2018-095410-B-I00); ISCIII-RETICs (RD16/0011/0030); Consejería de Salud y Familias, Junta de Andalucía (PIER-0084-2019) Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Efecto de 3,4-dihidroxifenilglicol, polifenol del aceite de oliva virgen extra, en la nefropatía diabética experimental.

Objetivo: El objetivo de este estudio fue analizar el efecto nefroprotector de 3',4'-dihidroxifenilglicol (DHPG), un compuesto polifenólico del aceite de oliva virgen extra (AOVE), sobre las lesiones renales en un modelo experimental de diabetes tipo 1. Material y Métodos: Estudio ex vivo mediante modelo experimental de Diabetes Mellitus tipo 1 en ratas Wistar. Grupos: ratas normoglucémicas (NDR), ratas diabéticas (DR) tratadas con solución salina, y DR tratadas con DHPG (0,5 mg/kg/día o 1 mg/kg/día). Valoración del estrés oxidativo: determinación de malondialdehído (MDA), capacidad antioxidante total, glutatión, 3-nitrotirosina, 8-isoprostano, 8-hidroxi-2-deoxiguanosina, 11-dehidrotromboxano B2 y 6-keto-prostaglandina F1, en suero, orina y tejido renal. Determinación del perfil renal (aclaramiento de creatinina) y las concentraciones de glucosa. Análisis morfométrico de secciones de riñón teñidas con hematoxilina-eosina y ácido peryódico de Schiff (PAS): volumen glomerular y glomeruloesclerosis. Resultados: Las DR mostraron un perfil de estrés oxidativo y nitrosativo sérico y renal significativamente mayor que las NDR, así como una menor producción de prostaciclina y un daño renal evidente (definido como excreción urinaria de proteínas, menor aclaramiento de creatinina, mayor volumen glomerular y mayor índice de glomeruloesclerosis). DHPG redujo el estrés oxidativo, nitrosativo (una reducción del 59,2% y del 34,7%, respectivamente, respecto a las DR no tratadas) y aumentó la producción de prostaciclina (7,8% en las ratas tratadas con DHPG). Así como, una reducción del 38-56% en la excreción urinaria de proteínas y del 22-46% en los parámetros morfológicos glomerulares (tras el tratamiento con 0,5 o 1 mg/kg/día, respectivamente).Conclusiones: La administración de DHPG a ratas diabéticas tipo 1 ejerce un efecto nefroprotector, probablemente debido a la suma de sus propiedades antioxidantes, antinitrosativas y regulador de la producción de prostaciclina.Universidad de Málaga. Campus de Excelencia

Plasma inorganic pyrophosphate and alkaline phosphatase in patients with pseudoxanthoma elasticum

Background: Inorganic pyrophosphate (PPi) plays a major role inhibiting dystrophic calcification. The aim was to analyze levels of PPi in patients having pseudoxanthoma elasticum (PXE), and controls as well as the enzymes who regulate the PPi plasma concentration. Methods: We collected fasting blood samples from PXE patients and age- and sex-matched controls in ethylenediamine tetraacetic acid (EDTA) and citrate-theophylline-adenosine-dipyridamole (CTAD) containing tubes. We measured PPi, ENPP1 mass and activity, alkaline phosphatase (AP) and tissue non-specific alkaline phosphatase (TNAP), CD73 and Human Platelet Factor-4 (CXCL4). Results: PPi in EDTA and CTAD samples were lower in PXE subjects than in controls (1.11±0.26 vs. 1.43±0.41 µM/L and 0.35±0.15 vs. 0.61±0.18 µM/L respectively, P<0.05). TNAP and liver TNAP activities were also higher in PXE than in controls (80.3±27.0 vs. 63.3±16.4 UI/L and 25.6±14.9 vs. 12.9±9.2 UI/L respectively, P<0.05). ENPP1 mass and activity as well as CD73 were almost identical. There was a weak but significant inverse correlation between TNAP activity and PPi levels (Pearson correlation −0.379, P<0.05) in both groups. Conclusions: High TNAP activity seems to contribute to low plasma levels of PPi in subjects with PXE, reinforcing the idea that pharmacological reduction of TNAP activity may help to reduce dystrophic calcification in PXE patients

Sporulation is dispensable for the vegetable-associated life cycle of the human pathogen Bacillus&nbsp;cereus.

Bacillus cereus is a common food-borne pathogen that is responsible for important outbreaks of food poisoning in humans. Diseases caused by B. cereus usually exhibit two major symptoms, emetic or diarrheic, depending on the toxins produced. It is assumed that after the ingestion of contaminated vegetables or processed food, spores of enterotoxigenic B. cereus reach the intestine, where they germinate and produce the enterotoxins that are responsible for food poisoning. In our study, we observed that sporulation is required for the survival of B. cereus in leaves but is dispensable in ready-to-eat vegetables, such as endives. We demonstrate that vegetative cells of B. cereus that are originally impaired in sporulation but not biofilm formation are able to reach the intestine and cause severe disorders in a murine model. Furthermore, our findings emphasise that the number of food poisoning cases associated with B. cereus is underestimated and suggest the need to revise the detection protocols, which are based primarily on spores and toxins

Sporulation is dispensable for the vegetable-associated life cycle of the human pathogen Bacillus&nbsp;cereus.

Bacillus cereus is a common food-borne pathogen that is responsible for important outbreaks of food poisoning in humans. Diseases caused by B. cereus usually exhibit two major symptoms, emetic or diarrheic, depending on the toxins produced. It is assumed that after the ingestion of contaminated vegetables or processed food, spores of enterotoxigenic B. cereus reach the intestine, where they germinate and produce the enterotoxins that are responsible for food poisoning. In our study, we observed that sporulation is required for the survival of B. cereus in leaves but is dispensable in ready-to-eat vegetables, such as endives. We demonstrate that vegetative cells of B. cereus that are originally impaired in sporulation but not biofilm formation are able to reach the intestine and cause severe disorders in a murine model. Furthermore, our findings emphasise that the number of food poisoning cases associated with B. cereus is underestimated and suggest the need to revise the detection protocols, which are based primarily on spores and toxins

Sporulation is dispensable for the vegetable-associated life cycle of the human pathogen Bacillus cereus

Bacillus cereus is a common food-borne pathogen that is responsible for important outbreaks of food poisoning in humans. Diseases caused by B. cereus usually exhibit two major symptoms, emetic or diarrheic, depending on the toxins produced. It is assumed that after the ingestion of contaminated vegetables or processed food, spores of enterotoxigenic B. cereus reach the intestine, where they germinate and produce the enterotoxins that are responsible for food poisoning. In our study, we observed that sporulation is required for the survival of B. cereus in leaves but is dispensable in ready-to-eat vegetables, such as endives. We demonstrate that vegetative cells of B. cereus that are originally impaired in sporulation but not biofilm formation are able to reach the intestine and cause severe disorders in a murine model. Furthermore, our findings emphasise that the number of food poisoning cases associated with B. cereus is underestimated and suggest the need to revise the detection protocols, which are based primarily on spores and toxins.This work was supported by grants from European Research Council Starting Grant (BacBio 637971), and Plan Nacional de I + D+i of Ministerio de Economía y Competitividad, Ministerio de Ciencia e Innovacion (AGL2016-78662-R and PID2019-107724GB-I00). Johan HJ Leveau was financed by USDA-NIFA grant number 2014-67017-21695. Juan Antonio Guadix acknowledges financial support by grant (PIER-0084-2019) from Proyectos de Investigacion en Salud de Junta de AndalucíaYe