PROTEINACEOUS COMPOUNDS FROM FRAGARIA ANANASSA FRUIT ATTENUATES PARAQUAT INDUCED PARKINSON LIKE LOCOMOTOR AND MITOCHONDRIAL ALTERATIONS IN ZEBRAFISH

Objectives: To assess the Parkinson like locomotor and mitochondrial alterations, associated with the exposure of paraquat (PQ), in vivo preventive effect of proteinaceous compounds extracted from Fragaria ananassa fruit (FA-D) against mitochondrial dysfunction induced by paraquat in zebra fish using brain mitochondria.Methods: Parkinson like locomotor and mitochondrial alterations were resulted by intra peritoneal administration of 55 mM PQ alternatively for a period of 7 days. The water soluble proteinaceous compounds from Fragaria ananassa fruit were obtained by Ammonium sulphate fractionation. The molecular weight of FA-D fraction was determined by SDS-PAGE and we found three distinct bands at 20.0 kDa, 17.0 kDa and 14.4.0 kDa bands respectively. The in vitro antioxidant activity and the in vivo preventive effect of FA-D against PQ induced Parkinsonian symptoms was evaluated by different assay systems viz., in vitro: radical scavenging activity by DPPH reaction and in vivo: locomotion, dopamine levels, complex-I activity, mitochondrial ROS levels, cytochrome c release and mitochondrial morphology.Results: The results show that paraquat altered locomotor activity and increased dopamine levels. Mitochondria isolated from paraquat treated zebrafish showed a marked inhibition of complex-I activity, increase in mitochondrial reactive oxygen species (mt ROS) and cytochrome c release and disintegration of mitochondrial structure. Treatment of 0.25 mg/kg body weight of FA-D fraction once in alternative days, for 5 days subsequent to the administration of PQ alternatively for a period of 7days, substantially reduced mt ROS levels and markedly restored the complex-I activity, cytochrome c release and mitochondrial morphology.Conclusion: The results strongly suggest that proteinaceous compounds from Fragaria ananassa fruit recuperate paraquat induced Parkinsonian like symptoms by protecting the mitochondria.Â

Long-term Transplant Function After Thrombolytic Treatment Ex Vivo of Donated Kidneys Retrieved 4 to 5 Hours After Circulatory Death

Background.\ua0Using a novel thrombolytic technique, we present long-term transplant function, measured by creatinine and iohexol clearance, after utilizing kidneys from porcine donors with uncontrolled donation after circulatory deaths, with 4.5–5 h of warm ischemia.Methods.\ua0Pigs in the study group were subjected to simulated circulatory death. After 2 h, ice slush was inserted into the abdomen and 4.5 h after death, the kidneys were retrieved. Lys-plasminogen, antithrombin-III, and alteplase were injected through the renal arteries on the back table. Subsequent ex vivo perfusion was continued for 3 h at 15\ub0C, followed by 3 h with red blood cells at 32\ub0C, and then transplanted into pigs as an autologous graft as only renal support. Living-donor recipient pigs that did not receive ex vivo perfusion, and unilateral nephrectomized pigs served as the controls.Results.\ua0Pigs in the study group (n = 13), surviving 10 d or more were included, of which 7 survived for 3 mo. Four animals in the living-donor group (n = 6) and all 5 nephrectomized animals survived for 3 mo. Creatinine levels in the plasma and urine, neutrophil gelatinase-associated lipocalin levels, Kidney Injury Marker-1 expression, and iohexol clearance at 3 mo did not differ significantly between the study and living-donor groups. Histology and transmission electron microscopy after 3 mo showed negligible fibrosis and no other damage.Conclusions.\ua0The present method salvages kidneys from extended unontrolled donation after circulatory death using thrombolytic treatment while preserving histology and enabling transplantation after ex vivo reconditioning, with clinically acceptable late function after 3 mo, as measured by creatinine and iohexol clearance

Characterization of Decellularized Implants for Extracellular Matrix Integrity and Immune Response Elicitation

Biological scaffold is a popular choice for the preparation of tissue-engineered organs and has the potential to address donor shortages in clinics. However, biological scaffolds prepared by physical or chemical agents cause damage to the extracellular matrix (ECM) by potentially inducing immune responses after implantation. The current study explores the fate of the decellularized (DC) scaffolds using a cocktail of chemicals following implantation without using immunosuppressants. Using the syngeneic (Lewis male-Lewis female) and allogeneic (Brown Norway male-Lewis female) models and different tissue routes (subcutaneous vs. omentum) for implantation, we applied in-depth quantitative proteomics, genomics along with histology and quantitative image analysis tools to comprehensively describe and compare the proteins following DC and postimplantation. Our data helped to identify any alteration postdecullarization as well implantation. We could also monitor route-specific modulation of the ECM and regulation of the immune responses (macrophage and T cells) following implantation. The current approach opens up the possibility to monitor the fate of biological scaffolds in terms of the ECM and immune response against the implants. In addition, the identification of different routes helped us to identify differential immune responses against the implants. This study opens up the potential to identify the changes associated with chemical DC both pre- and postimplantation, which could further help to promote research in this direction. The development of a biological scaffold helps in the preparation of a functional organ in the clinics. In the current study, we develop a strategy for chemical decellularization and explored two different routes to understand the differential responses elicited postimplantation. The use of sensitive protein and genomic tools to study the changes creates a favorable environment for similar efforts to develop and characterize biological scaffolds before further trials in the clinics. The current study, which was carried out without any immunosuppressive agents, could help to establish (a) appropriate chemical strategies for preparing biological scaffolds as well as (b) identify putative implantable routes to circumvent any adverse immune reactions, which will ultimately decide the outcome for acceptance or rejection of the scaffold/implant

Proceedings Of The 23Rd Paediatric Rheumatology European Society Congress: Part Two

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