In vitro and in vivo evaluation of AFB1 and OTA-toxicity through immunofluorescence and flow cytometry techniques : A systematic review

Due to the globalization, mycotoxins have been considered a major risk to human health being the main con- taminants of foodstuffs. Among them, AFB1 and OTA are the most toxic and studied. Therefore, the goal of this review is to deepen the knowledge about the toxicological effects that AFB1 and OTA can induce on human health by using flow cytometry and immunofluorescence techniques in vitro and in vivo models. The examination of the selected reports shows that the majority of them are focused on immunotoxicity while the rest are con- cerned about nephrotoxicity, hepatotoxicity, gastrointestinal toxicity, neurotoxicity, embryotoxicity, reproduc- tive system, breast, esophageal and lung toxicity. In relation to immunofluorescence analysis, biological processes related to AFB1- and OTA-toxicity were evaluated such as inflammation, neuronal differentiation, DNA damage, oxidative stress and cell death. In flow cytometry analysis, a wide range of assays have been performed across the reviewed studies being apoptosis assay, cell cycle analysis and intracellular ROS measurement the most employed. Although, the toxic effects of AFB1 and OTA have been reported, further research is needed to clarify AFB1 and OTA-mechanism of action on human health

Transcriptional changes after enniatins A, A1, B and B1 ingestion in rat stomach, liver, kidney and lower intestine

Enniatins (ENs) are depsipeptide mycotoxins produced by Fusarium fungi. They are known for their capacity to modulate cell membrane permeability and disruption of ionic gradients, affecting cell homeostasis and initiating oxidative stress mechanisms. The effect of the acute toxicity of ENs A, A1, B and B1 at two different concentrations after 8 h of exposure was analysed in Wistar rats by a transcriptional approach. The following key mitochondrial and nuclear codified genes related to the electron transport chain were considered for gene expression analysis in stomach, liver, kidney and lower intestine by quantitative Real-Time PCR: mitochondrially encoded NADH dehydrogenase 1 (MT-ND1), mitochondrially encoded cytochrome c oxidase 1 (MT-COX1), succinate dehydrogenase flavoprotein subunit A and ATP synthase F1 subunit alpha, respectively. Moreover, the expression of markers involved in oxidative stresssuperoxide dismutase 1 (SOD1), glutathione peroxidase 1 (Gpx1), heme oxygenase 1, apoptosis B-cell lymphoma 2, Bcl2 Associated protein X (Bax), tumor suppressor protein (p53), inhibition of apoptosis nuclear factor kappa of activated B cells, immune system interleukin 1β and intestinal tight junction Occludin merely in lower intestine tissues have been investigated. For mitochondrial genes, the main differences were observed for MT-ND1 and MT-COX1, showing its deficiency in all selected organs. With regard to the intestinal barrier's cellular response to oxidative stress, the activity of the antioxidant gene SOD1 was decreased in a dose-dependent manner. Similarly, the catalytic enzyme GPx1 was also downregulated though merely at medium dose employed. On the contrary, the pro-apoptotic Bax and p53 regulators were activated after ENs exposure, reporting a significant increase in their expression. Furthermore, the alteration of intestinal permeability was assessed by the abnormal activity of the tight junction protein occludin. In summary, ENs may generate mitochondrial disorders and induce oxidative stress in intestinal barrier function

Transcriptional profiling reveals functional links between RasGrf1 and Pttg1 in pancreatic beta cells

Background: Our prior characterization of RasGrf1 deficient mice uncovered significant defects in pancreatic islet count and size as well as beta cell development and signaling function, raising question about the mechanisms linking RasGrf1 to the generation of those"pancreatic" phenotypes. Results: Here, we compared the transcriptional profile of highly purified pancreatic islets from RasGrf1 KO mice to that of WT control animals using commercial oligonucleotide microarrays. RasGrf1 elimination resulted in differential gene expression of numerous components of MAPK- and Calcium-signaling pathways,suggesting a relevant contribution of this GEF to modulation of cellular signaling in the cell lineages integrating the pancreatic islets. Whereas the overall transcriptional profile of pancreatic islets was highly specific in comparison to other organs of the same KO mice, a significant specific repression of Pttg1 was a common transcriptional alteration shared with other tissues of neuroectodermal origin. This observation,together with the remarkable pancreatic phenotypic similarities between RasGrf1 KO and Pttg1 KO mice suggested the possibility of proximal functional regulatory links between RasGrf1 and Pttg1 in pancreatic cell lineages expressing these proteins. Analysis of the mPttg1 promoter region identified specific recognition sites for numerous transcription factors which were also found to be differentially expressed in RasGrf1 KO pancreatic islets and are known to be relevant for Ras-ERK signaling as well as beta cell function. Reporter luciferase assays in BT3 insulinoma cells demonstrated the ability of RasGrf1 to modulate mPttg1 promoter activity through ERK-mediated signals. Analysis of the phenotypic interplay between RasGrf1 and Pttg1 in double knockout RasGrf1/Pttg1 mice showed that combined elimination of the two loci resulted in dramatically reduced values of islet and beta cell count and glucose homeostasis function which neared those measured in single Pttg1 KO mice and were significantly lower than those observed in individual RasGrf1 KO mice. Conclusions: The specific transcriptional profile and signaling behavior of RasgGrf1 KO pancreatic islets, together with the dominance of Pttg1 over RasGrf1 with regards to the generation of these phenotypes in mouse pancreas, suggest that RasGrf1 is an important upstream component of signal transduction pathways regulating Pttg1 expression and controlling beta cell development and physiological response

Transcriptional profiling reveals functional links between RasGrf1 and Pttg1 in pancreatic beta cells

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License .[Background]: Our prior characterization of RasGrf1 deficient mice uncovered significant defects in pancreatic islet count and size as well as beta cell development and signaling function, raising question about the mechanisms linking RasGrf1 to the generation of those >pancreatic> phenotypes. [Results]: Here, we compared the transcriptional profile of highly purified pancreatic islets from RasGrf1 KO mice to that of WT control animals using commercial oligonucleotide microarrays. RasGrf1 elimination resulted in differential gene expression of numerous components of MAPK- and Calcium-signaling pathways, suggesting a relevant contribution of this GEF to modulation of cellular signaling in the cell lineages integrating the pancreatic islets. Whereas the overall transcriptional profile of pancreatic islets was highly specific in comparison to other organs of the same KO mice, a significant specific repression of Pttg1 was a common transcriptional alteration shared with other tissues of neuroectodermal origin. This observation, together with the remarkable pancreatic phenotypic similarities between RasGrf1 KO and Pttg1 KO mice suggested the possibility of proximal functional regulatory links between RasGrf1 and Pttg1 in pancreatic cell lineages expressing these proteins. [Conclusions]: The specific transcriptional profile and signaling behavior of RasgGrf1 KO pancreatic islets, together with the dominance of Pttg1 over RasGrf1 with regards to the generation of these phenotypes in mouse pancreas, suggest that RasGrf1 is an important upstream component of signal transduction pathways regulating Pttg1 expression and controlling beta cell development and physiological responses.Work supported by grants FIS PI13/02846 and RTICC RD12/0036/0001 from Instituto de Salud Carlos III (ISCIII), and grant SA181U13 from JCyL, Spain. We are grateful to Dr. Douglas Lowy (National Cancer Institute, Bethesda, MD) for providing plasmid pBK-CMV RasGrf1 and to Dr. Shlomo Melmed (Cedars-Sinai Medical Center, Los Angeles, CA) for providing reporter plasmid pGL3-Pttg1 and the single PTTG1 KO mouse strain used in these studies.Peer Reviewe

Estudios clínicos sobre la enfermedad celíaca (2014-2019): revisión sistemática de la prevalencia de la presentación clínica y enfermedades asociadas por edades

Introduction: Celiac disease (CD) is characterized by a wide variety of signs, symptoms and associated diseases in its presentation, it can even be asymptomatic. Recent studies show the variation of the clinical spectrum according to age. In young children, classic celiac disease predominates with symptoms such as abdominal distension, decreased appetite, diarrhea and weight loss. Frequent atypical manifestations in older children are abdominal pain, constipation, reflux, vomiting, fatigue and short stature. In adults there is a reduction in the classical presentation and an increase in the non-classical.Material and methods: A systematic review was conducted in which articles on signs and symptoms of classical and atypical CD in children and adults have been included, in addition to the diseases that are frequently associated with this pathology. The databases used to search the articles were PubMed, Web of Science and Scopus, from the last six years (2014-2019). A total of 164 articles have been evaluated following the selection criteria, of which the 20 most relevant articles have been included in the study.Results: CD is diagnosed more frequently in women and at earlier ages than men. In children under two years old, classical CD predominates although more intensely than in young children. In older children, teenagers and adults, the form of presentation changes towards atypical or non-classical and asymptomatic.Conclusions: The increased prevalence in the last years, great heterogeneity of symptoms, associated diseases and the variation of the clinical spectrum towards an atypical form with extraintestinal symptoms causes delay in its diagnosis so they must be recognized to be detected earlier.Introducción: La enfermedad celíaca (EC) se caracteriza por una gran variedad de signos, síntomas y enfermedades asociadas en su forma de presentación, incluso puede cursar de forma asintomática. Recientes estudios muestran la variación del espectro clínico según la edad. En niños y niñas pequeñas predomina la forma clásica con síntomas como distensión abdominal, disminución del apetito, diarrea y pérdida de peso. Las manifestaciones atípicas frecuentes en niños mayores son dolor abdominal, estreñimiento, reflujo, vómitos, fatiga, talla baja. En adultos se observa una reducción de la forma de presentación clásica e incremento de la no clásica.Material y métodos: Se realizó una revisión sistemática en la que se han incluido los artículos sobre signos y síntomas de la EC clásica y atípica en niños y adultos, además de las enfermedades que se asocian de forma frecuente a esta patología. Las bases de datos utilizadas para la búsqueda de los artículos fueron PubMed, Web of Science y Scopus, de los últimos seis años (2014-2019). Se han evaluado un total de 164 artículos tras los criterios de selección, de los cuales se han incluido en el estudio los 20 artículos más relevantes.Resultados: La EC se diagnostica con mayor frecuencia en mujeres y a edades más tempranas que a los hombres. En niños menores de dos años predomina la EC clásica, aunque de forma más intensa que en niños y niñas pequeñas. En niños y niñas mayores, adolescentes y edad adulta varía la forma de presentación hacia la atípica o no clásica y asintomática.Conclusiones: El incremento en la prevalencia en los últimos años, gran heterogeneidad de síntomas, enfermedades asociadas y la variación del espectro clínico hacia una forma atípica con síntomas extraintestinales causa retraso en su diagnóstico por lo que deben reconocerse para que se detecte de manera más precoz

Bioaccessibility study of aflatoxin B1 and ochratoxin A in bread enriched with fermented milk whey and/or pumpkin

The presence of mycotoxins in cereals and cereal products remains a significant issue. The use of natural ingredients such as pumpkin and whey, which contain bioactive compounds, could be a strategy to reduce the use of conventional chemical preservatives. The aim of the present work was to study the bioaccessibility of aflatoxin B1 (AFB1) and ochratoxin (OTA) in bread, as well as to evaluate the effect of milk whey (with and without lactic acid bacteria fermentation) and pumpkin on reducing mycotoxins bioaccessibility. Different bread typologies were prepared and subjected to an in vitro digestion model. Gastric and intestinal extracts were analyzed by HPLC-MS/qTOF and mycotoxins bioaccessibility was calculated. All the tested ingredients but one significantly reduced mycotoxin intestinal bioaccessibility. Pumpkin powder demonstrated to be the most effective ingredient showing significant reductions of AFB1 and OTA bioaccessibility up to 74% and 34%, respectively. Whey, fermented whey, and the combination of pumpkin-fermented whey showed intestinal bioaccessibility reductions between 57-68% for AFB1, and between 11-20% for OTA. These results pointed to pumpkin and milk whey as potential bioactive ingredients that may have promising applications in the bakery industry

Multi-occurrence of twenty mycotoxinsin pasta and a risk assessment in the moroccan population

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified quick, easy, cheap effective, rugged, and safe method was validated. The mycotoxins studied were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS). The validated method was applied to one hundred and six (n = 106) pasta samples purchased from several areas in the country. The analytical results showed that 99 out of 106 total samples (93.4%) were contaminated with at least one mycotoxin. Nine mycotoxins (Aflatoxin B1, Enniatin B, Enniatin B1, Enniatin A1, Zearalenone, Deoxynivalenol, 3-Acetyl-Deoxynivalenol, T-2, and HT-2 toxins) were present in the pasta samples. Enniatin B and Enniatin B1 were the predominant mycotoxins. The Zearalenone, Deoxynivalenol, HT-2, and T-2 toxins were present in 51.8%, 43.5%, 34.9%, and 16% of samples, respectively. Aflatoxin B1 was detected in only 2 samples. Risk exposure assessment concluded that mycotoxin levels found in pasta do not pose a significant human health risk for the Moroccan population. This is the first paper drafted on the multi-occurrence of mycotoxins in pasta from this countr

Uloga karotenoida iz ekstrakta mesa bundeve u zaštiti od oštećenja krvno-moždane barijere uzrokovane mikotoksinima in vitro

Some mycotoxins such as beauvericin (BEA), ochratoxin A (OTA), and zearalenone (ZEA) can cross the blood brain barrier, which is why we tested the anti-inflammatory action of a pumpkin carotenoid extract (from the pulp) against these mycotoxins and their combinations (OTA+ZEA and OTA+ZEA+BEA) on a blood brain barrier model with co-cultured ECV304 and C6 cells using an untargeted metabolomic approach. The cells were added with mycotoxins at a concentration of 100 nmol/L per mycotoxin and pumpkin carotenoid extract at 500 nmol/L. For control we used only vehicle solvent (cell control) or vehicle solvent with pumpkin extract (extract control). After two hours of exposure, samples were analysed with HPLC-ESI-QTOF-MS. Metabolites were identified against the Metlin database. The proinflammatory arachidonic acid metabolite eoxin (14,15-LTE4) showed lower abundance in ZEA and BEA+OTA+ZEA-treated cultures that also received the pumpkin extract than in cultures that were not treated with the extract. Another marker of inflammation, prostaglandin D2-glycerol ester, was only found in cultures treated with OTA+ZEA and BEA+OTA+ZEA but not in the ones that were also treated with the pumpkin extract. Furthermore, the concentration of the pumpkin extract metabolite dihydromorelloflavone significantly decreased in the presence of mycotoxins. In conclusion, the pumpkin extract showed protective activity against cellular inflammation triggered by mycotoxins thanks to the properties pertinent to flavonoids contained in the pulp.Pojedini mikotoksini poput bovericina (BEA), okratoksina A (OTA) i zearalenona (ZEA) prelaze krvno-moždanu barijeru, a to je i razlog zbog kojega smo istražili djelovanje ekstrakta karotenoida iz mesa bundeve protiv upalnih procesa izazvanih ovim mikotoksinima i njihovim kombinacijama (OTA+ZEA i OTA+ZEA+BEA) na modelu krvno-moždane barijere koji se sastojao od kultura stanica ECV304 i C6, oslanjajući se pritom na neciljani metabolomički pristup. Stanice su tretirane mikotoksinima u koncentraciji od 100 nmol/L po mikotoksinu odnosno ekstraktom karotenoida u koncentraciji od 500 nmol/L. Za kontrolu smo upotrijebili samo otapalo (stanična kontrola) odnosno otapalo s bundevinim ekstraktom (ekstraktna kontrola). Nakon dva sata tretmana uzorci su analizirani metodom tekućinske kromatografije / masene spektrometrije (HPLC-ESI-QTOF-MS), a dobiveni metaboliti identificirani su usporedbom s bazom podataka Metlin. Primjena ekstrakta značajno je smanjila količinu metabolita proupalne arahidonske kiseline eoksina (14,15-LTE4) u kulturama tretiranima samo zearalenonom odnosno kombinacijom BEA+OTA+ZEA. Drugi upalni biljeg, prostaglandin D2-glicerol ester, otkriven je samo u kulturama tretiranima kombinacijama OTA+ZEA odnosno BEA+OTA+ZEA, ali ne i u onima koje su usto tretirane bundevinim ekstraktom. Osim toga, u prisutnosti mikotoksina značajno je porasla koncentracija metabolita dihidromoreloflavona iz bundevina ekstrakta. Time je taj ekstrakt iskazao zaštitno djelovanje protiv stanične upale uzrokovane mikotoksinima zahvaljujući svojstvima flavonoida koji se nalaze u njezinu mesu

Implicación de PTTG1 en procesos celulares y fisiológicos controlados por RASGRF1

Memoria presentada por la licenciada Lara Manyes i Font para optar al grado de Doctora por la Universidad de Salamanca y realizada en Instituto de Biología Molecular y Celular del Cancer de Salamanca.[EN]: In the laboratory of Eugene Santos, using profile analysis dependent transcriptional Rasgrf1 expression in mouse retina, was that among the genes most affected by deficiency in Rasgrf1 was PTTG1. Another analysis of the same kind in this laboratory, both in mouse islets and in cerebral cortex and olfactory bulb, resubmitted to PTTG1 as affected by gene Rasgrf1 deficiency. The main objective of this thesis was to study whether controlled Rasgrf1 PTTG1 expression. The second was to analyze whether the in vivo functions of PTTG1 Rasgrf1 and overlapped. The generation of stable cell clones by shRNA PC12 allowed us to study the proliferative rate, and cell cycle status with reduced intracellular signaling Rasgrf1 and PTTG1 expression. We conducted luciferase assays to examine the promoter activity PTTG1. To study the in vivo function of these proteins generated RasGrf1-/--Pttg1-/- mice. We employed the Barnes maze to analyze its role in hippocampus, where both are expressed PTTG1 as Rasgrf1 mostly. Also described that both mice PTTG1 Rasgrf1 KO ​​KO as look decreased body mass relative to control mice suffer insulinopenia and have a mass reduced beta cell, so do the test glucose tolerance double KO mice. In summary, these results suggest a role for the control Rasgrf1 expression PTTG1, via the signaling path of Mek. Rasgrf1 and PTTG1 could share overlapping functions pancreatic level, but not at the level of the central nervous system, where only PTTG1 seems to have a role in learning and hippocampal-dependent memory.[ES]: En el laboratorio de Eugenio Santos, mediante análisis del perfil transcripcional dependiente de la expresión de RasGrf1 en retina de ratón, se vio que entre los genes mas afectados por la deficiencia en RasGrf1 estaba Pttg1. Otro análisis del mismo tipo realizado en este laboratorio, tanto en islotes de ratón como en córtex cerebral y bulbo olfatorio, volvió a presentar a Pttg1 como gen afectado por la deficiencia de RasGrf1. El objetivo principal de esta tesis fue estudiar si RasGrf1 controlaba la expresión de Pttg1. El segundo fue analizar si las funciones in vivo de RasGrf1 y Pttg1 se solapaban. La generación de clones estables de células PC12 mediante shRNA nos permitió estudiar la tasa proliferativa, el ciclo celular y el estado de la señalización intracelular con una menor expresión de RasGrf1 y Pttg1. Llevamos a cabo ensayos de luciferasa para examinar la actividad del promotor de Pttg1. Para el estudio de la función in vivo de estas proteínas generamos ratones RasGrf1-/--Pttg1-/-. Hemos empleado el laberinto de Barnes para el análisis de su función en hipocampo, donde tanto Pttg1 como RasGrf1 se expresan mayoritariamente. Asimismo, se ha descrito que tanto los ratones RasGrf1 KO como los Pttg1 KO ven disminuida su masa corporal con respecto a los ratones control, sufren insulinopenia y tienen una masa de células ß reducida, por lo que realizamos el test de tolerancia a la glucosa a los ratones doble KO. En resumen, estos resultados sugieren un papel para RasGrf1 en el control de la expresión de Pttg1, a través de la ruta de señalización de Mek. RasGrf1 y Pttg1 podrían compartir funciones solapadas a nivel pancreático, pero no a nivel del sistema nervioso central, dónde sólo Pttg1 parece tener una función en el aprendizaje y memoria dependiente de hipocampo.Esta memoria ha sido realizada siendo LARA MANYES I FONT beneficiaria de una ayuda predoctoral FPU del Ministerio de Educación para la elaboración de la tesis doctoral (2008‐2012). La investigación en el laboratorio ha sido financiada por los siguientes proyectos: ‐ Red Temática de Investigación Cooperativa en Cáncer, Centro de Investigación del Cáncer de Salamanca‐IBMCC (CSIC‐USAL). Fundación de Investigación del Cáncer (Cod. 1556). Ministerio de Sanidad. Fondo de Investigación Sanitaria (2007‐2009). IP, E. Santos. ‐ Mecanismos de activación de oncoproteínas Ras: Análisis de especificidad funcional de diana Ras y sus activadores GEF en procesos fisiológicos y patológicos. Ministerio de Sanidad. Fondo de Investigación Sanitaria (2007‐ 2009). IP, E. Santos. ‐ Estudios de especificidad funcional de oncoproteínas Ras y sus activadores celulares. Junta de Castilla y León (SA044A08) (2008‐2010). IP, E. Santos. ‐ Especificidad funcional de proteínas Ras y sus activadores GEF en procesos fisiológicos y patológicos. Junta de Castilla y León GR93, Grupos de excelencia de Castilla y León (2008‐2010). IP, E. Santos. ‐ Mecanismos de especificidad funcional de proteínas Ras y sus activadores celulares específicos GEF en procesos fisiológicos y patológicos. Fondo de Investigaciones Sanitarias. (Proyecto Intrasalud cPS09/01979) (2010‐2013). IP, E. Santos.Peer Reviewe

Implicación de PTTG1 en procesos celulares y fisiológicos controlados por RASGRF1

[ES] En el laboratorio de Eugenio Santos, mediante análisis del perfil transcripcional dependiente de la expresión de RasGrf1 en retina de ratón, se vio que entre los genes mas afectados por la deficiencia en RasGrf1 estaba Pttg1. Otro análisis del mismo tipo realizado en este laboratorio, tanto en islotes de ratón como en córtex cerebral y bulbo olfatorio, volvió a presentar a Pttg1 como gen afectado por la deficiencia de RasGrf1. El objetivo principal de esta tesis fue estudiar si RasGrf1 controlaba la expresión de Pttg1. El segundo fue analizar si las funciones in vivo de RasGrf1 y Pttg1 se solapaban. La generación de clones estables de células PC12 mediante shRNA nos permitió estudiar la tasa proliferativa, el ciclo celular y el estado de la señalización intracelular con una menor expresión de RasGrf1 y Pttg1. Llevamos a cabo ensayos de luciferasa para examinar la actividad del promotor de Pttg1. Para el estudio de la función in vivo de estas proteínas generamos ratones RasGrf1-/--Pttg1-/-. Hemos empleado el laberinto de Barnes para el análisis de su función en hipocampo, donde tanto Pttg1 como RasGrf1 se expresan mayoritariamente. Asimismo, se ha descrito que tanto los ratones RasGrf1 KO como los Pttg1 KO ven disminuida su masa corporal con respecto a los ratones control, sufren insulinopenia y tienen una masa de células ß reducida, por lo que realizamos el test de tolerancia a la glucosa a los ratones doble KO. En resumen, estos resultados sugieren un papel para RasGrf1 en el control de la expresión de Pttg1, a través de la ruta de señalización de Mek. RasGrf1 y Pttg1 podrían compartir funciones solapadas a nivel pancreático, pero no a nivel del sistema nervioso central, dónde sólo Pttg1 parece tener una función en el aprendizaje y memoria dependiente de hipocampo.[EN] In the laboratory of Eugene Santos, using profile analysis dependent transcriptional Rasgrf1 expression in mouse retina, was that among the genes most affected by deficiency in Rasgrf1 was PTTG1. Another analysis of the same kind in this laboratory, both in mouse islets and in cerebral cortex and olfactory bulb, resubmitted to PTTG1 as affected by gene Rasgrf1 deficiency. The main objective of this thesis was to study whether controlled Rasgrf1 PTTG1 expression. The second was to analyze whether the in vivo functions of PTTG1 Rasgrf1 and overlapped. The generation of stable cell clones by shRNA PC12 allowed us to study the proliferative rate, and cell cycle status with reduced intracellular signaling Rasgrf1 and PTTG1 expression. We conducted luciferase assays to examine the promoter activity PTTG1. To study the in vivo function of these proteins generated RasGrf1-/--Pttg1-/- mice. We employed the Barnes maze to analyze its role in hippocampus, where both are expressed PTTG1 as Rasgrf1 mostly. Also described that both mice PTTG1 Rasgrf1 KO ​​KO as look decreased body mass relative to control mice suffer insulinopenia and have a mass reduced beta cell, so do the test glucose tolerance double KO mice. In summary, these results suggest a role for the control Rasgrf1 expression PTTG1, via the signaling path of Mek. Rasgrf1 and PTTG1 could share overlapping functions pancreatic level, but not at the level of the central nervous system, where only PTTG1 seems to have a role in learning and hippocampal-dependent memory