[Streptokinase: extraction, cloning and overexpression of recombinant protein with simplified purification technique]

Streptokinase has been widely prescribed as a fibrinolytic agent for myocardial infarction. Simple structural characteristics of this protein have provided techniques for production of different recombinant types of this protein. The present study was designed to prepare equisimilisH46A subtype of streptokinase in our country. Having extracted the DNA, the streptokinase gene was replicated and cloned in pGEX-4T-2. The recombinant product was transformed in BL21 [DE3] plysS'. Then the recombinant streptokinase expression and performance was assessed by lasic densitometry and special test for S2251 substrate. Use of a restriction enzyme for both sides of a gene may facilitate its cloning in different expressive carriers. Expression rate of recombinant protein [45%] confirmed successful cloning. Use of pGEX-4T-2 carrier was not only associated with active recombinant streptokinase production, added GST to its amine ending that could facilitate purification process

Testis specific gene 10 expression in the testes of patients with non-obstructive azoospermia

Disorders in the expression of any gene effective in spermatogenic pathway is known as a probable cause of non-obstructive azoospermia and male infertility. The way responsible genes for sperm motility are expressed can considerably affect male fertility. Recent studies show that TSGA10 gene is effective in the natural process of spermatogenesis as protein produced by this gene in mouse results in the production of the main structure of sperm tail. Up to now, no comprehensive studies have been done on the way this gene is expressed in the infertile's testical tissue. In this study, TSGA10 mRNA expression in testicular samples of 84 patients with non-obstructive azoospermia was investigated by semi-quantitative nested RT-PCR in Avesina Infertility Clinic during 2005-6. Moreover, expression levels of TSGA10 during spermatogenesis were evaluated using Johnsen's method for histopathological scoring of the samples. For statistical analysis, SPSS software [Version 11.2] was used. The difference between gene expressions was done based on quantitative variables by the use of t-test and covariance analysis and alpha<0.05 was regarded as a statistically significant value. Testicular TSGA10 mRNA expression was observed in 31 patients, [36.9%], with non-obstructive azoospermia which it had a statistically significant correlation with spermatogenesis progress [p<000.0]. Histopathologically, the gene had been expressed in patients with higher Johnsen's score of spermatogenesis while a lack of expression was seen in all of those with Johnsen's score less than 4.5. The findings indicate that TSGA10 is expressed in human testis and it is restricted to germ cells. It seems that lack of TSGA10 expression may have negative effects on spermatogenesis and on male fertility. On the other hand, determination of the timing of gene expression in a certain level of spermatogenesis may also be used to determine levels of spermatogenesis in azoospermic patients alongside histopathological findings

case report of recurrent hydrops fetalis due to maternal alloimmunization with Kell antigen

During pregnancy, irregular blood group antibodies originating either from earlier pregnancies or from blood transfusions may severely affect child health. In this report, a case of maternal alloimmunization to Kell antigen is described.The mother had a history of partial mole and four repeated intrauterine fetal death due to hydrops fetalis.Screening of irregular blood group antibodies revealed that she has anti-Kell with the titer of 1:4096. Also in genetic analysis, a C677T homozygous mutation of MTHFR gene was found, which could potentially enhance destructive effects of anti-Kell antibody. The described case emphasizes the importance of being informed about the presence of irregular blood group antibodies during pregnancy which may cause recurrent hydrops