Background:Undifferentiated shock is recognized as a criticality state that is transitional in immune-mediated topology for casual risk of lethal microcirculatory dysfunction.This was a sensitivity analysis of a drug(...Background:Undifferentiated shock is recognized as a criticality state that is transitional in immune-mediated topology for casual risk of lethal microcirculatory dysfunction.This was a sensitivity analysis of a drug(tetracosactide;TCS10)targeting melanocortin receptors(MCRs)in a phase 3 randomized controlled trial to improve cardiovascular surgical rescue outcome by reversing mortality and hemostatic disorders.Methods:Sensitivity analysis was based on a randomized,two-arm,multicenter,double-blind,controlled trial.The Naïve Bayes classifier was performed by density-based sensitivity index for principal strata as proportional hazard model of 30-day surgical risk mortality according to European System for Cardiac Operative Risk Evaluation inputs outputs in 100 consecutive cases(from August to September 2013 from Emilia Romagna region,Italy).Patients included an agent-based TCS10 group(10 mg,single intravenous bolus before surgery;n=56)and control group(n=44)and the association with cytokines,lactate,and bleeding-blood transfusion episodes with the prior-risk log odds for mortality rate in time-to-event was analyzed.Results:Thirty-day mortality was significantly improved in the TCS10 group vs.control group(0 vs.8 deaths,P<0.0001).Baseline levels of interleukin(IL)-6,IL-10,and lactate were associated with bleeding episodes,independent of TCS10 treatment[odds ratio(OR)=1.90,95%confidence interval(CI)1.39-2.79;OR=1.53,95%CI 1.17-2.12;and OR=2.92,95%CI 1.40-6.66,respectively],while baseline level of Fms-like tyrosine kinase 3 ligand(Flt3L)was associated with lower bleeding rates in TCS10-treated patients(OR=0.31,95%CI 0.11-0.90,P=0.03).For every 8 TCS10-treated patients,1 bleeding case was avoided.Blood transfusion episodes were significantly reduced in the TCS10 group compared to the control group(OR=0.32,95%CI 0.14-0.73,P=0.01).For every 4 TCS10-treated patients,1 transfusion case was avoided.Conclusions:Sensitivity index underlines the quality target product profile of TCS10 in the runway of emergency casualty care.To introduce the technology readiness level in real-life critically ill patients,further large-scale studies are required.Trial registration:European Union Drug Regulating Authorities Clinical Trials Database(EudraCT Number:2007-006445-41).展开更多
AIM: To develop an in vitro model based on neural stem cells derived from transgenic animals, to be used in the study of pathological mechanisms of Alzheimer's disease and for testing new molecules.METHODS: Neural...AIM: To develop an in vitro model based on neural stem cells derived from transgenic animals, to be used in the study of pathological mechanisms of Alzheimer's disease and for testing new molecules.METHODS: Neural stem cells(NSCs) were isolated from the subventricular zone of Wild type(Wt) and Tg2576 mice. Primary and secondary neurosphere generation was studied, analysing population doubling and the cell yield per animal. Secondary neurospheres were dissociated and plated on MCM Gel Cultrex 2D and after 6 d in vitro(DIVs) in mitogen withdrawal conditions,spontaneous differentiation was studied using specific neural markers(MAP2 and TuJ-1 for neurons, GFAP forastroglial cells and CNPase for oligodendrocytes). Gene expression pathways were analysed in secondary neurospheres, using the QIAGEN PCR array for neurogenesis, comparing the Tg2576 derived cell expression with the Wt cells. Proteins encoded by the altered genes were clustered using STRING web software.RESULTS: As revealed by 6E10 positive staining, all Tg2576 derived cells retain the expression of the human transgenic Amyloid Precursor Protein. Tg2576 derived primary neurospheres show a decrease in population doubling. Morphological analysis of differentiated NSCs reveals a decrease in MAP2- and an increase in GFAP-positive cells in Tg2576 derived cells. Analysing the branching of TuJ-1 positive cells, a clear decrease in neurite number and length is observed in Tg2576 cells.The gene expression neurogenesis pathway revealed11 altered genes in Tg2576 NSCs compared to Wt.CONCLUSION: Tg2576 NSCs represent an appropriate AD in vitro model resembling some cellular alterations observed in vivo, both as stem and differentiated cells.展开更多
Myelination,remyelination and demyelination:modeling the in vitro drug discovery pipeline:Demyelination is a multifactorial event occurring in diseases primarily involving myelin forming cells(oligodendrocytes,OLs)and...Myelination,remyelination and demyelination:modeling the in vitro drug discovery pipeline:Demyelination is a multifactorial event occurring in diseases primarily involving myelin forming cells(oligodendrocytes,OLs)and their precursors(oligodendrocyte precursor cells,OPCs)such as multiple sclerosis,but is also involved in the pathology of other central nervous system(CNS)injuries and diseases,such as neonatal encephalopathy。展开更多
基金funded by the National Plan Military Research (EF a2011.188)
文摘Background:Undifferentiated shock is recognized as a criticality state that is transitional in immune-mediated topology for casual risk of lethal microcirculatory dysfunction.This was a sensitivity analysis of a drug(tetracosactide;TCS10)targeting melanocortin receptors(MCRs)in a phase 3 randomized controlled trial to improve cardiovascular surgical rescue outcome by reversing mortality and hemostatic disorders.Methods:Sensitivity analysis was based on a randomized,two-arm,multicenter,double-blind,controlled trial.The Naïve Bayes classifier was performed by density-based sensitivity index for principal strata as proportional hazard model of 30-day surgical risk mortality according to European System for Cardiac Operative Risk Evaluation inputs outputs in 100 consecutive cases(from August to September 2013 from Emilia Romagna region,Italy).Patients included an agent-based TCS10 group(10 mg,single intravenous bolus before surgery;n=56)and control group(n=44)and the association with cytokines,lactate,and bleeding-blood transfusion episodes with the prior-risk log odds for mortality rate in time-to-event was analyzed.Results:Thirty-day mortality was significantly improved in the TCS10 group vs.control group(0 vs.8 deaths,P<0.0001).Baseline levels of interleukin(IL)-6,IL-10,and lactate were associated with bleeding episodes,independent of TCS10 treatment[odds ratio(OR)=1.90,95%confidence interval(CI)1.39-2.79;OR=1.53,95%CI 1.17-2.12;and OR=2.92,95%CI 1.40-6.66,respectively],while baseline level of Fms-like tyrosine kinase 3 ligand(Flt3L)was associated with lower bleeding rates in TCS10-treated patients(OR=0.31,95%CI 0.11-0.90,P=0.03).For every 8 TCS10-treated patients,1 bleeding case was avoided.Blood transfusion episodes were significantly reduced in the TCS10 group compared to the control group(OR=0.32,95%CI 0.14-0.73,P=0.01).For every 4 TCS10-treated patients,1 transfusion case was avoided.Conclusions:Sensitivity index underlines the quality target product profile of TCS10 in the runway of emergency casualty care.To introduce the technology readiness level in real-life critically ill patients,further large-scale studies are required.Trial registration:European Union Drug Regulating Authorities Clinical Trials Database(EudraCT Number:2007-006445-41).
基金Supported by Regione Emilia Romagna,POR-FESR 2011-2014
文摘AIM: To develop an in vitro model based on neural stem cells derived from transgenic animals, to be used in the study of pathological mechanisms of Alzheimer's disease and for testing new molecules.METHODS: Neural stem cells(NSCs) were isolated from the subventricular zone of Wild type(Wt) and Tg2576 mice. Primary and secondary neurosphere generation was studied, analysing population doubling and the cell yield per animal. Secondary neurospheres were dissociated and plated on MCM Gel Cultrex 2D and after 6 d in vitro(DIVs) in mitogen withdrawal conditions,spontaneous differentiation was studied using specific neural markers(MAP2 and TuJ-1 for neurons, GFAP forastroglial cells and CNPase for oligodendrocytes). Gene expression pathways were analysed in secondary neurospheres, using the QIAGEN PCR array for neurogenesis, comparing the Tg2576 derived cell expression with the Wt cells. Proteins encoded by the altered genes were clustered using STRING web software.RESULTS: As revealed by 6E10 positive staining, all Tg2576 derived cells retain the expression of the human transgenic Amyloid Precursor Protein. Tg2576 derived primary neurospheres show a decrease in population doubling. Morphological analysis of differentiated NSCs reveals a decrease in MAP2- and an increase in GFAP-positive cells in Tg2576 derived cells. Analysing the branching of TuJ-1 positive cells, a clear decrease in neurite number and length is observed in Tg2576 cells.The gene expression neurogenesis pathway revealed11 altered genes in Tg2576 NSCs compared to Wt.CONCLUSION: Tg2576 NSCs represent an appropriate AD in vitro model resembling some cellular alterations observed in vivo, both as stem and differentiated cells.
基金the ARSEP Foundation(Fondation pour l’aideàla recherchésur la sclérose en plaques)for its support in developing the in NSC-derived OPC in vitro systems as part of the“Role of RXRγin T3-mediated oligodendrocyte differentiation and remyelination”project(to LC and VAB)and fellowship(to VAB)。
文摘Myelination,remyelination and demyelination:modeling the in vitro drug discovery pipeline:Demyelination is a multifactorial event occurring in diseases primarily involving myelin forming cells(oligodendrocytes,OLs)and their precursors(oligodendrocyte precursor cells,OPCs)such as multiple sclerosis,but is also involved in the pathology of other central nervous system(CNS)injuries and diseases,such as neonatal encephalopathy。
