Wildlife DNAForensic isthe application of regular DNAforensic methodsfor proper identification of wildlife parts and their products.Recent advances in molecular genetic studies have generated a new and exciting range ...Wildlife DNAForensic isthe application of regular DNAforensic methodsfor proper identification of wildlife parts and their products.Recent advances in molecular genetic studies have generated a new and exciting range of possible applications of genetic methods to wildlife research,conservation,and management.These advances have led to an explosion in genetic research on wildlife for their identification at molecular level and have increased interest among researchers working in other scientific disciplines for application of genetic technology in wildlife DNAforensic field.Different molecular markers have been developed and being routinely used for analysis,such as nuclear markers(variable number of tandem repeats,single-nucleotide polymorphisms),mitochondrial markers(cytochrome b,cytochrome c oxidase subunit I,16S rRNA,12S rRNA,and D‑Loop)and microsatellites.As soon as,a case is reported under Wildlife Protection Act(1972)the case exhibits are sent to forensic laboratories for proper analysis of species for appropriate application of law.展开更多
Introduction:Storage of specimens sampled from human remains for pathological testing,embalming for burial purposes,and for human identification often requires formalin fixation and/or paraffin embedding.Current knowl...Introduction:Storage of specimens sampled from human remains for pathological testing,embalming for burial purposes,and for human identification often requires formalin fixation and/or paraffin embedding.Current knowledge in molecular biology techniques and forensic DNA analysis makes it possible to optimize the extraction of amplifiable DNA from formalin-fixed tissues by improving the pre-treatment,optimizing the digestion condition of proteinase K,simplifying the extraction protocol and purifying the extracted DNA with optimized volumes of alcohol.Aim:This research sought to extract amplifiable DNA from thirteen brain,bone marrow and cartilage samples from four formalin embalmed human cadavers.Materials and Methods:Brain,cartilage and bone marrow samples were taken from four different cadavers at autopsy at the Ghana Police Hospital mortuary in Accra,Ghana sixty-two days after embalming.An optimized preparation and DNA extraction protocol was carried out on all the samples.Brain samples were also taken from a non-formalin treated fifth cadaver of known STR profile,and standard DNA extraction performed to serve as positive control.Results:Our optimized protocol yielded detectable quantities of DNA from the samples when quantified with the 7500 Real-Time PCR equipment.The extracted DNA also yielded full STR profiles with varying peak heights for forensic identification purposes.The measured degradation indexes of the DNA samples were greater than 1.0,with peak heights of generated STR profiles above the limits of detection of the 3500 genetic analyzer.Conclusion:Our current study demonstrated an optimized method of DNA extraction from tissues(brain,cartilage and bone marrow)sampled from formalin embalmed human cadavers.The optimized protocol reduced the concentration of formalin fixation residues in extracted DNA from formalin-fixed tissues,thereby improving the amplification efficiency for STR profiling.Brain,bone marrow and cartilages can be a good source of DNA from embalmed and degraded human remains,though for skeletonized human remains together with teeth and long bones.展开更多
Plant extracts intended for application as natural food antioxidants are largely touted as safe antioxidants without mechanistic interpretation of their in vivo efficacy or tangible proof of their toxicological effect...Plant extracts intended for application as natural food antioxidants are largely touted as safe antioxidants without mechanistic interpretation of their in vivo efficacy or tangible proof of their toxicological effects.The study used RNA-Seq analysis to evaluate the transcriptomic effects of Harpephyllum caffrum Berhn.Fruit peel extracts(HCP)at 250,450 and 650 ppm in Saccharomyces cerevisiae and their ability to attenuate in vivo H_(2)O_(2)-induced oxidative stress in comparison with sodium metabisulphite(SMB)at 450 ppm.The HCP were influenced ribosome biogenesis and spliceosome relative to SMB.Additionally,HCP treatments primed cells for oxidative stress response and recovery with HCP-450 application being optimal,while SMB induced differential upregulation of thioredoxin,peroxiredoxin,glutathione peroxidase and catalase oxidative stress response genes.There were signs of mitochondrial damage in stressed HCP-650,while the proteasomal system and pro-apoptotic genes were induced in the stressed SMB treatment.Overall,the study demonstrates the antioxidant efficacy of HCP,especially at 450 ppm,and its potential to induce epitranscriptomic modifications beneficial for a healthful diet as opposed to SMB.展开更多
文摘Wildlife DNAForensic isthe application of regular DNAforensic methodsfor proper identification of wildlife parts and their products.Recent advances in molecular genetic studies have generated a new and exciting range of possible applications of genetic methods to wildlife research,conservation,and management.These advances have led to an explosion in genetic research on wildlife for their identification at molecular level and have increased interest among researchers working in other scientific disciplines for application of genetic technology in wildlife DNAforensic field.Different molecular markers have been developed and being routinely used for analysis,such as nuclear markers(variable number of tandem repeats,single-nucleotide polymorphisms),mitochondrial markers(cytochrome b,cytochrome c oxidase subunit I,16S rRNA,12S rRNA,and D‑Loop)and microsatellites.As soon as,a case is reported under Wildlife Protection Act(1972)the case exhibits are sent to forensic laboratories for proper analysis of species for appropriate application of law.
文摘Introduction:Storage of specimens sampled from human remains for pathological testing,embalming for burial purposes,and for human identification often requires formalin fixation and/or paraffin embedding.Current knowledge in molecular biology techniques and forensic DNA analysis makes it possible to optimize the extraction of amplifiable DNA from formalin-fixed tissues by improving the pre-treatment,optimizing the digestion condition of proteinase K,simplifying the extraction protocol and purifying the extracted DNA with optimized volumes of alcohol.Aim:This research sought to extract amplifiable DNA from thirteen brain,bone marrow and cartilage samples from four formalin embalmed human cadavers.Materials and Methods:Brain,cartilage and bone marrow samples were taken from four different cadavers at autopsy at the Ghana Police Hospital mortuary in Accra,Ghana sixty-two days after embalming.An optimized preparation and DNA extraction protocol was carried out on all the samples.Brain samples were also taken from a non-formalin treated fifth cadaver of known STR profile,and standard DNA extraction performed to serve as positive control.Results:Our optimized protocol yielded detectable quantities of DNA from the samples when quantified with the 7500 Real-Time PCR equipment.The extracted DNA also yielded full STR profiles with varying peak heights for forensic identification purposes.The measured degradation indexes of the DNA samples were greater than 1.0,with peak heights of generated STR profiles above the limits of detection of the 3500 genetic analyzer.Conclusion:Our current study demonstrated an optimized method of DNA extraction from tissues(brain,cartilage and bone marrow)sampled from formalin embalmed human cadavers.The optimized protocol reduced the concentration of formalin fixation residues in extracted DNA from formalin-fixed tissues,thereby improving the amplification efficiency for STR profiling.Brain,bone marrow and cartilages can be a good source of DNA from embalmed and degraded human remains,though for skeletonized human remains together with teeth and long bones.
基金supported by the South African Research Chairs Initia-tive(SARChI)in Meat Science:Genomics to Nutrinomics and partly funded by the South African Department of Science and Innovation(UID:84633),as directed by the NRF of South AfricaTMP is thankful to the SARChI for bursary support+1 种基金TMP would also like to acknowledge Stellenbosch University Central Analytical Facility(CAF)and Alvera Vorster for performing the RNA-seq workflow and DIPLOMICS for the financial support via the Quality Control training related subsidyThe authors appreciate Dr J.K.Marima for her assistance with analysis and interpretation training.
文摘Plant extracts intended for application as natural food antioxidants are largely touted as safe antioxidants without mechanistic interpretation of their in vivo efficacy or tangible proof of their toxicological effects.The study used RNA-Seq analysis to evaluate the transcriptomic effects of Harpephyllum caffrum Berhn.Fruit peel extracts(HCP)at 250,450 and 650 ppm in Saccharomyces cerevisiae and their ability to attenuate in vivo H_(2)O_(2)-induced oxidative stress in comparison with sodium metabisulphite(SMB)at 450 ppm.The HCP were influenced ribosome biogenesis and spliceosome relative to SMB.Additionally,HCP treatments primed cells for oxidative stress response and recovery with HCP-450 application being optimal,while SMB induced differential upregulation of thioredoxin,peroxiredoxin,glutathione peroxidase and catalase oxidative stress response genes.There were signs of mitochondrial damage in stressed HCP-650,while the proteasomal system and pro-apoptotic genes were induced in the stressed SMB treatment.Overall,the study demonstrates the antioxidant efficacy of HCP,especially at 450 ppm,and its potential to induce epitranscriptomic modifications beneficial for a healthful diet as opposed to SMB.
