Data-driven drug repositioning using olfactory omics profiles-challenges and perspectives in neurodegeneration:Neurodegenerative diseases are characterized by progressive degeneration and loss of neuronal function in ...Data-driven drug repositioning using olfactory omics profiles-challenges and perspectives in neurodegeneration:Neurodegenerative diseases are characterized by progressive degeneration and loss of neuronal function in the central nervous system.These diseases are often characterized as proteinopathies,which are disorders primarily driven by the aggregation or misfolding of specific amyloid proteins within cells,leading to their dysfunction and eventual death.Despite the gain-of-function hypothesis related to the aggregation of these proteins,recently,an alternative hypothesis regarding the loss-of-function of the soluble monomeric proteins during the process of aggregation into amyloids is gaining currency.This last event is called proteinopenia and refers to conditions characterized by a deficiency or decrease in the levels of specific soluble proteins in the body(Ezzat et al.,2023).It has been demonstrated that levels of soluble proteins involved in neurodegenerative diseases are decreased.展开更多
BACKGROUND Arachidyl amido cholanoic acid(Aramchol)is a potent downregulator of hepatic stearoyl-CoA desaturase 1(SCD1)protein expression that reduces liver triglycerides and fibrosis in animal models of steatohepatit...BACKGROUND Arachidyl amido cholanoic acid(Aramchol)is a potent downregulator of hepatic stearoyl-CoA desaturase 1(SCD1)protein expression that reduces liver triglycerides and fibrosis in animal models of steatohepatitis.In a phase IIb clinical trial in patients with nonalcoholic steatohepatitis(NASH),52 wk of treatment with Aramchol reduced blood levels of glycated hemoglobin A1c,an indicator of glycemic control.AIM To assess lipid and glucose metabolism in mouse hepatocytes and in a NASH mouse model[induced with a 0.1%methionine and choline deficient diet(0.1MCD)]after treatment with Aramchol.METHODS Isolated primary mouse hepatocytes were incubated with 20μmol/L Aramchol or vehicle for 48 h.Subsequently,analyses were performed including Western blot,proteomics by mass spectrometry,and fluxomic analysis with 13C-uniformly labeled glucose.For the in vivo part of the study,male C57BL/6J mice were randomly fed a control or 0.1MCD for 4 wk and received 1 or 5 mg/kg/d Aramchol or vehicle by intragastric gavage for the last 2 wk.Liver metabolomics were assessed using ultra-high-performance liquid chromatography-time of flight-MS for the determination of glucose metabolism-related metabolites.RESULTS Combination of proteomics and Western blot analyses showed increased AMPK activity while the activity of nutrient sensor mTORC1 was decreased by Aramchol in hepatocytes.This translated into changes in the content of their downstream targets including proteins involved in fatty acid(FA)synthesis and oxidation[PACCα/β(S79),SCD1,CPT1A/B,HADHA,and HADHB],oxidative phosphorylation(NDUFA9,NDUFB11,NDUFS1,NDUFV1,ETFDH,and UQCRC2),tricarboxylic acid(TCA)cycle(MDH2,SUCLA2,and SUCLG2),and ribosome(P-p70S6K[T389]and P-S6[S235/S236]).Flux experiments with 13Cuniformely labeled glucose showed that TCA cycle cataplerosis was reduced by Aramchol in hepatocytes,as indicated by the increase in the number of rounds that malate remained in the TCA cycle.Finally,liver metabolomic analysis showed that glucose homeostasis was improved by Aramchol in 0.1MCD fed mice in a dose-dependent manner,showing normalization of glucose,G6P,F6P,UDP-glucose,and Rbl5P/Xyl5P.CONCLUSION Aramchol exerts its effect on glucose and lipid metabolism in NASH through activation of AMPK and inhibition of mTORC1,which in turn activate FAβ-oxidation and oxidative phosphorylation.展开更多
Nowadays, at a time of growing concern for sustainable development and compliance with environmental standards and legislation, the detection of heavy metal contaminants in environmental matrices represents a difficul...Nowadays, at a time of growing concern for sustainable development and compliance with environmental standards and legislation, the detection of heavy metal contaminants in environmental matrices represents a difficult but important task. The current major limitation lies in the poor detection limits of the targeted pollutant's trace concentrations by the available conventional techniques. In order to elaborate a novel "living" self assembled electrochemical 3-D biosensor, the authors propose a new concept to overcome this shortcoming. The advantages of the properties of polyelectrolyte-functionalized NBs (nanobeads) are combined along with the use of non covalently strongly bound micro-organisms. The designed 3-D biosensor is all the more promising as it has showed a significantly improved sensitivity. In fact, the detection limits of the tested heavy metals (cadmium and mercury) were as low as 1.0 × 10^-12 mol.L-1 and six to seven orders of magnitude lower than those provided by conventional 2-D biosensors. Furthermore, it is potentially applicable to a wide range of bioreceptor-pollutant detection systems.展开更多
Climate change scenarios predict an increase in mean air temperatures and in the frequency,intensity,and length of extreme temperature events in many wine-growing regions worldwide.Because elevated temperature has det...Climate change scenarios predict an increase in mean air temperatures and in the frequency,intensity,and length of extreme temperature events in many wine-growing regions worldwide.Because elevated temperature has detrimental effects on berry growth and composition,it threatens the economic and environmental sustainability of wine production.Using Cabernet Sauvignon fruit-bearing cuttings,we investigated the effects of high temperature(HT)on grapevine berries through a label-free shotgun proteomic analysis coupled to a complementary metabolomic study.Among the 2,279 proteins identified,592 differentially abundant proteins were found in berries exposed to HT.The gene ontology categories“stress,”“protein,”“secondary metabolism,”and“cell wall”were predominantly altered under HT.High temperatures strongly impaired carbohydrate and energy metabolism,and the effects depended on the stage of development and duration of treatment.Transcript amounts correlated poorly with protein expression levels in HT berries,highlighting the value of proteomic studies in the context of heat stress.Furthermore,this work reveals that HT alters key proteins driving berry development and ripening.Finally,we provide a list of differentially abundant proteins that can be considered as potential markers for developing or selecting grape varieties that are better adapted to warmer climates or extreme heat waves.展开更多
Background:Human corneal stromal keratocytes propagated in culture media supplemented with human amnion extract(AME)can correct early corneal haze in an animal model.Clinical application of cultivated keratocytes is l...Background:Human corneal stromal keratocytes propagated in culture media supplemented with human amnion extract(AME)can correct early corneal haze in an animal model.Clinical application of cultivated keratocytes is limited by infectious disease screening before amnion products can be used in humans.It remains unclear if AME from cryopreserved versus fresh human amnion can support human keratocyte propagation,and which components of the extract promote keratocyte growth.Methods:Three placentas were collected for the preparation of fresh and cryopreserved amnion tissues followed by homogenization and protein extraction.AME protein profiles were studied using isobaric tagging for relative and absolute quantitation(iTRAQ)proteomics.Enriched gene ontology(GO)terms and functional classes were identified.Primary human keratocytes from 4 donor corneas were cultured in media supplemented with fresh AME(F-AME)or cryopreserved AME(C-AME).Cell viability,proliferation and keratocyte marker expression were examined by confocal immunofluorescence and flow cytometry.Results:AME proteomics revealed 1385 proteins with similar expression levels(between 0.5-and 2-fold)between Fand C-AME,while 286 proteins were reduced(less than 0.5-fold)in C-AME.Enriched GO term and biological pathway analysis showed that those proteins with comparable expression between F-AME and C-AME were involved in cell metabolism,epithelial-mesenchymal transition,focal adhesion,cell-extracellular matrix interaction,cell stress regulation and complement cascades.Human corneal stromal keratocytes cultured with F-AME or C-AME showed similar morphology and viability,while cell proliferation was mildly suppressed with C-AME(P>0.05).Expression of aldehyde dehydrogenase 3A1(ALDH3A1)and CD34 was similar in both cultures.Conclusion:AME from cryopreserved amnion had limited influence on keratocyte culture.It is feasible to use protein extract from cryopreserved amnion to propagate human keratocytes for potential translational applications.展开更多
P2X7 purinoceptor promotes survival or cytotoxicity depending on extraceUular adenosine triphosphate (ATP) stimulus Intensity controlling its ion channel or P2X7-dependent large pore (LP) functions. Mechanisms gov...P2X7 purinoceptor promotes survival or cytotoxicity depending on extraceUular adenosine triphosphate (ATP) stimulus Intensity controlling its ion channel or P2X7-dependent large pore (LP) functions. Mechanisms governing this operational divergence and functional idiosyncrasy are ill-understood. We have discovered a feedback loop where sustained activation of P2X7 triggers release of active matrix metalloproteinase 2 (MMP-2), which halts ion channel and LP responses via the MMP-2-dependent receptor cleavage. This mechanism operates in cells as diverse as macrophages, dystrophic myoblasts, P2X7-transfected HEK293, and human tumour cells. Given that serum-born MMP-2 activity also blocked receptor functions, P2X7 responses in vivo may decrease in organs with permeable capillaries. Therefore, this mechanism represents an Important fine-tuning of P2X7 functions, reliant on both cell-autonomous and extraneous factors. Indeed, it allowed evasion from the ATP-induced cvtotoxicity in macrophages and human cancer ceils with high P2X7 expression levels. Finally, we demonstrate that P2X7 ablation eliminated getatinase activity in inflamed dystrophic muscles in vivo. Thus, P2X7 antagonists could be used as an alternative to highly toxic MMP inhibitors in treatments of inflammatory diseases and cancers.展开更多
基金funded by grants from the Spanish Ministry of Science,Innovation and Universities(Ref.PID2019-110356RB-I00/AEI/10.13039/501100011033)to JFI and ESthe Department of Economic and Business Development from Government of Navarra(Ref.0011-1411-2023-000028 to ES)+2 种基金supported by a predoctoral fellowship from the Public University of Navarra(UPNA)supported by a postdoctoral fellowship from Miguel Servet Foundation-Navarrabiomedsupported by“Programa MRR Investigo 2023”in the framework of the European Union recovery and resilience facility。
文摘Data-driven drug repositioning using olfactory omics profiles-challenges and perspectives in neurodegeneration:Neurodegenerative diseases are characterized by progressive degeneration and loss of neuronal function in the central nervous system.These diseases are often characterized as proteinopathies,which are disorders primarily driven by the aggregation or misfolding of specific amyloid proteins within cells,leading to their dysfunction and eventual death.Despite the gain-of-function hypothesis related to the aggregation of these proteins,recently,an alternative hypothesis regarding the loss-of-function of the soluble monomeric proteins during the process of aggregation into amyloids is gaining currency.This last event is called proteinopenia and refers to conditions characterized by a deficiency or decrease in the levels of specific soluble proteins in the body(Ezzat et al.,2023).It has been demonstrated that levels of soluble proteins involved in neurodegenerative diseases are decreased.
基金National Institutes of Health Grant,No.R01CA172086Plan Nacional of I+D,No.SAF2017-88041-R+5 种基金Ministerio de Economía y Competitividad de España,No.SAF2017-87301-RAsociación Española contra el Cáncer,No.AECC17/302Ayudas Fundación BBVA a equipos de Investigación Científica 2018Fondo Europeo de Desarrollo Regional,Ministerio de Economia y Competitividad de España,No.PGC2018-099857-BI00Basque Government Grants,No.IT1264-19Ministerio de Economia y Competitividad de España for the Severo Ochoa Excellence Accreditation,No.SEV-2016-0644.The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘BACKGROUND Arachidyl amido cholanoic acid(Aramchol)is a potent downregulator of hepatic stearoyl-CoA desaturase 1(SCD1)protein expression that reduces liver triglycerides and fibrosis in animal models of steatohepatitis.In a phase IIb clinical trial in patients with nonalcoholic steatohepatitis(NASH),52 wk of treatment with Aramchol reduced blood levels of glycated hemoglobin A1c,an indicator of glycemic control.AIM To assess lipid and glucose metabolism in mouse hepatocytes and in a NASH mouse model[induced with a 0.1%methionine and choline deficient diet(0.1MCD)]after treatment with Aramchol.METHODS Isolated primary mouse hepatocytes were incubated with 20μmol/L Aramchol or vehicle for 48 h.Subsequently,analyses were performed including Western blot,proteomics by mass spectrometry,and fluxomic analysis with 13C-uniformly labeled glucose.For the in vivo part of the study,male C57BL/6J mice were randomly fed a control or 0.1MCD for 4 wk and received 1 or 5 mg/kg/d Aramchol or vehicle by intragastric gavage for the last 2 wk.Liver metabolomics were assessed using ultra-high-performance liquid chromatography-time of flight-MS for the determination of glucose metabolism-related metabolites.RESULTS Combination of proteomics and Western blot analyses showed increased AMPK activity while the activity of nutrient sensor mTORC1 was decreased by Aramchol in hepatocytes.This translated into changes in the content of their downstream targets including proteins involved in fatty acid(FA)synthesis and oxidation[PACCα/β(S79),SCD1,CPT1A/B,HADHA,and HADHB],oxidative phosphorylation(NDUFA9,NDUFB11,NDUFS1,NDUFV1,ETFDH,and UQCRC2),tricarboxylic acid(TCA)cycle(MDH2,SUCLA2,and SUCLG2),and ribosome(P-p70S6K[T389]and P-S6[S235/S236]).Flux experiments with 13Cuniformely labeled glucose showed that TCA cycle cataplerosis was reduced by Aramchol in hepatocytes,as indicated by the increase in the number of rounds that malate remained in the TCA cycle.Finally,liver metabolomic analysis showed that glucose homeostasis was improved by Aramchol in 0.1MCD fed mice in a dose-dependent manner,showing normalization of glucose,G6P,F6P,UDP-glucose,and Rbl5P/Xyl5P.CONCLUSION Aramchol exerts its effect on glucose and lipid metabolism in NASH through activation of AMPK and inhibition of mTORC1,which in turn activate FAβ-oxidation and oxidative phosphorylation.
文摘Nowadays, at a time of growing concern for sustainable development and compliance with environmental standards and legislation, the detection of heavy metal contaminants in environmental matrices represents a difficult but important task. The current major limitation lies in the poor detection limits of the targeted pollutant's trace concentrations by the available conventional techniques. In order to elaborate a novel "living" self assembled electrochemical 3-D biosensor, the authors propose a new concept to overcome this shortcoming. The advantages of the properties of polyelectrolyte-functionalized NBs (nanobeads) are combined along with the use of non covalently strongly bound micro-organisms. The designed 3-D biosensor is all the more promising as it has showed a significantly improved sensitivity. In fact, the detection limits of the tested heavy metals (cadmium and mercury) were as low as 1.0 × 10^-12 mol.L-1 and six to seven orders of magnitude lower than those provided by conventional 2-D biosensors. Furthermore, it is potentially applicable to a wide range of bioreceptor-pollutant detection systems.
基金This research received funding from the Agence Nationale de la Recherche for the project"DURAVITIS"(ANR-2010-GENM-004-01).
文摘Climate change scenarios predict an increase in mean air temperatures and in the frequency,intensity,and length of extreme temperature events in many wine-growing regions worldwide.Because elevated temperature has detrimental effects on berry growth and composition,it threatens the economic and environmental sustainability of wine production.Using Cabernet Sauvignon fruit-bearing cuttings,we investigated the effects of high temperature(HT)on grapevine berries through a label-free shotgun proteomic analysis coupled to a complementary metabolomic study.Among the 2,279 proteins identified,592 differentially abundant proteins were found in berries exposed to HT.The gene ontology categories“stress,”“protein,”“secondary metabolism,”and“cell wall”were predominantly altered under HT.High temperatures strongly impaired carbohydrate and energy metabolism,and the effects depended on the stage of development and duration of treatment.Transcript amounts correlated poorly with protein expression levels in HT berries,highlighting the value of proteomic studies in the context of heat stress.Furthermore,this work reveals that HT alters key proteins driving berry development and ripening.Finally,we provide a list of differentially abundant proteins that can be considered as potential markers for developing or selecting grape varieties that are better adapted to warmer climates or extreme heat waves.
基金Singapore National Eye Centre HREF 0618-2Singapore National Research Foundation under Clinician Scientist Award-Senior Investigator Category(JRNMRR163801)National Medical Research Council,Ministry of Health,Singapore.
文摘Background:Human corneal stromal keratocytes propagated in culture media supplemented with human amnion extract(AME)can correct early corneal haze in an animal model.Clinical application of cultivated keratocytes is limited by infectious disease screening before amnion products can be used in humans.It remains unclear if AME from cryopreserved versus fresh human amnion can support human keratocyte propagation,and which components of the extract promote keratocyte growth.Methods:Three placentas were collected for the preparation of fresh and cryopreserved amnion tissues followed by homogenization and protein extraction.AME protein profiles were studied using isobaric tagging for relative and absolute quantitation(iTRAQ)proteomics.Enriched gene ontology(GO)terms and functional classes were identified.Primary human keratocytes from 4 donor corneas were cultured in media supplemented with fresh AME(F-AME)or cryopreserved AME(C-AME).Cell viability,proliferation and keratocyte marker expression were examined by confocal immunofluorescence and flow cytometry.Results:AME proteomics revealed 1385 proteins with similar expression levels(between 0.5-and 2-fold)between Fand C-AME,while 286 proteins were reduced(less than 0.5-fold)in C-AME.Enriched GO term and biological pathway analysis showed that those proteins with comparable expression between F-AME and C-AME were involved in cell metabolism,epithelial-mesenchymal transition,focal adhesion,cell-extracellular matrix interaction,cell stress regulation and complement cascades.Human corneal stromal keratocytes cultured with F-AME or C-AME showed similar morphology and viability,while cell proliferation was mildly suppressed with C-AME(P>0.05).Expression of aldehyde dehydrogenase 3A1(ALDH3A1)and CD34 was similar in both cultures.Conclusion:AME from cryopreserved amnion had limited influence on keratocyte culture.It is feasible to use protein extract from cryopreserved amnion to propagate human keratocytes for potential translational applications.
文摘P2X7 purinoceptor promotes survival or cytotoxicity depending on extraceUular adenosine triphosphate (ATP) stimulus Intensity controlling its ion channel or P2X7-dependent large pore (LP) functions. Mechanisms governing this operational divergence and functional idiosyncrasy are ill-understood. We have discovered a feedback loop where sustained activation of P2X7 triggers release of active matrix metalloproteinase 2 (MMP-2), which halts ion channel and LP responses via the MMP-2-dependent receptor cleavage. This mechanism operates in cells as diverse as macrophages, dystrophic myoblasts, P2X7-transfected HEK293, and human tumour cells. Given that serum-born MMP-2 activity also blocked receptor functions, P2X7 responses in vivo may decrease in organs with permeable capillaries. Therefore, this mechanism represents an Important fine-tuning of P2X7 functions, reliant on both cell-autonomous and extraneous factors. Indeed, it allowed evasion from the ATP-induced cvtotoxicity in macrophages and human cancer ceils with high P2X7 expression levels. Finally, we demonstrate that P2X7 ablation eliminated getatinase activity in inflamed dystrophic muscles in vivo. Thus, P2X7 antagonists could be used as an alternative to highly toxic MMP inhibitors in treatments of inflammatory diseases and cancers.