Cytotoxic Effect of Gliotoxin, Hemolysin, Protease and Melanin purified from Aspergillus fumigates on REF Cell Line, in vitro Study

Aspergillus fumigates especially clinical isolate produces a series of toxic substances and proteinaceous hemolysin, protease and pigment like melanin which appear to act in an additive and synergic way on cells. In this study, gliotoxin, hemolysin, Protease, and Melanin were used in an experimental model to study their Cytotoxic activity by evaluating their effect on REF cell line (Rat embryonic fibroblast ), for exposure time of 24 hrs at three different concentrations of each compounds triplicate of each concentration were used, cytotoxicity of the purified compounds were active against REF cell line under study and a toxic effect was clear with a significant difference at the level of probability (p≤ 0.05) and this effect was contrasted among different concentrations for each purified compound growth inhibition of REF cell line was increase gradually with the increase of compound concentration

Using Random Amplified Polymorphic DNA (RAPD) analysis to investigation of genetic diversity, and relationships among a set of clinical Aspergillus fumigatus isolates

This study is an attempt to determine the genetic diversity and relationships among fourteen local isolate isolated from patients with Aspergillosis (Aspergillus fumigatus) by using the Random Amplified Polymorphic DNA (RAPD) technique. Twelve universal primers used in this study produced 94 bands across fourteen isolates. Of these bands, 67 bands or 71.2% were polymorphic. The size of the amplified bands ranged between 100-2000 bp. The genetic polymorphism value of each primer was determined and ranged between 33-100%. In terms of unique banding patterns, determine the finger print for six isolates the most characteristic banding pattern was for the (AFU1, AFU2, AFU3, AFU4, AFU8 and AFU14) with primer (OP F-16 , OP I-06, OP F-16, OP X-01, OP X-01and OP A-06). Genetic distances ranged from 0.12419 to 0.64404 among A. fumigatus isolates. Cluster analyses were performed to construct a dendrogram among studied A. fumigatus isolates. The cluster analysis places most of the A.fumigatus isolates isolated from patient come from yhe same area into a close relation (subcluster) showing a high level of genetic relatedness and were distinct from isolates from another area (the other subcluster). Interestingly, a number of isolates originating from the same sources did form well defined groups, indicating association between the RAPD patterns and the geographic origin of the isolates. The information generated from this study can be used in the future for controlling of Aspergillosis programs

Cytotoxic Effect of Gliotoxin, Hemolysin, Protease and Melanin purified from Aspergillus fumigates on REF Cell Line, in vitro Study

Aspergillus fumigates especially clinical isolate produces a series of toxic substances and proteinaceous hemolysin, protease and pigment like melanin which appear to act in an additive and synergic way on cells. In this study, gliotoxin, hemolysin, Protease, and Melanin were used in an experimental model to study their Cytotoxic activity by evaluating their effect on REF cell line (Rat embryonic fibroblast ), for exposure time of 24 hrs at three different concentrations of each compounds triplicate of each concentration were used, cytotoxicity of the purified compounds were active against REF cell line under study and a toxic effect was clear with a significant difference at the level of probability (p≤ 0.05) and this effect was contrasted among different concentrations for each purified compound growth inhibition of REF cell line was increase gradually with the increase of compound concentration.</jats:p

Using Random Amplified Polymorphic DNA (RAPD) analysis to investigation of genetic diversity, and relationships among a set of clinical Aspergillus fumigatus isolates

This study is an attempt to determine the genetic diversity and relationships among fourteen local isolate isolated from patients with Aspergillosis (Aspergillus fumigatus) by using the Random Amplified Polymorphic DNA (RAPD) technique. Twelve universal primers used in this study produced 94 bands across fourteen isolates. Of these bands, 67 bands or 71.2% were polymorphic. The size of the amplified bands ranged between 100-2000 bp. The genetic polymorphism value of each primer was determined and ranged between 33-100%. In terms of unique banding patterns, determine the finger print for six isolates the most characteristic banding pattern was for the (AFU1, AFU2, AFU3, AFU4, AFU8 and AFU14) with primer (OP F-16 , OP I-06, OP F-16, OP X-01, OP X-01and OP A-06). Genetic distances ranged from 0.12419 to 0.64404 among A. fumigatus isolates. Cluster analyses were performed to construct a dendrogram among studied A. fumigatus isolates. The cluster analysis places most of the A.fumigatus isolates isolated from patient come from yhe same area into a close relation (subcluster) showing a high level of genetic relatedness and were distinct from isolates from another area (the other subcluster). Interestingly, a number of isolates originating from the same sources did form well defined groups, indicating association between the RAPD patterns and the geographic origin of the isolates. The information generated from this study can be used in the future for controlling of Aspergillosis programs.</jats:p