[Analysis of beta thalassemia mutations using the single strand conformation polymorphism [SSCP] technique]

beta-thalassemia [beta-thal] is one of the most prevalent hereditary diseases in Iran. There are more than two million carriers of beta-thal in Iran. Detection of the beta globin gene mutations is necessary for a definitive diagnostic and management plan such as prenatal diagnosis of beta-thalassemia. In our country, the PCR-Amplification Refractory Mutation System [PCR-ARMS] has been frequently used for detection of beta globin gene mutations. Here, we used the PCR-single strand conformation polymorphism [PCR-SSCP] assay for detection of mutations of beta globin gene. In the patients with confirmed mutations, we amplified 281base pairs containing exon of one of a beta globin gene by PCR. Based on SSCP technique 2.5 micro l of the reaction products appeared in polyacryamide gel electrophoresis and the bands were visualized by silver staining. Seven mutations and one polymorphism were evaluated by PCR SSCP assay. The results of this study demonstrated that the patterns of mobility of single strands were different from each other and those of control sample. Our study showed the PCR-SSCP technique can meet the need for direct genomic sequencing of DNA and could be applied in the developing countries where financial resources are limited but genetic hemoglobin disorders are highly prevalent

[Detection of beta-thalassemia mutations in Kurd patients of Kurdistan and West Azerbaijan provinces]

Beta-thalassemia is the most common autosomal recessive disorder. More than 200 different mutations in the beta-globin gene have been detected which can lead to decreased or absent beta-globin chain synthesis. Since the Iranian Population is a mixture of different ethnic groups, it is necessary to determine the frequency and distribution of these mutations in the different ethnic groups of our county. Therefore, in this study we determined the Spectrum and the frequency of beta-thalassemia mutations in the patients with beta-thalassemia major in the Kurd population of Kurdistan and West Azerbaijan provinces of Iran. To detect mutations, extracted DNA of 110 chromosomes from 55 unrelated patients, were studied by PCR-ARMS [Polymerase Chain Reaction-Amplification Refractory Mutation System] SSCP [Single Strand Conformation Polymorphism] and direct sequencing methods. The results of this study showed that IVS-II-1 [G-A] was the most common mutation with a frequency of 31%; FSC 8/9[+G] with a frequency of 19% was the second most prevalent mutation among all chromosomes. Other mutations were IVS-I-1[G-A] FSC8 [-AA] IVS-I-110[G-A] FSC36/37[-T] IVS-I-5[G-C], IVS-I-128[T-G] FSC44 [-C], FSC 5[-CT] and +22UTR [G-A] These mutations comprised 79% of beta-thalassemia mutations in this region and 21% of the mutations still remains to be explored. The results of this study showed that, there are similarities and differences between this region and other parts of Iran and also neighboring countries. Therefore, determination of beta-thalassemia mutations in this region seems to be necessary and beneficial for designing prenatal diagnosis programs

[Detection of rare beta globin gene mutations in north western Iran]

Recent molecular studies on Iranian -thalassemia genes revealed the presence of eight common mutations associated with beta-thalassemia. Although these mutations are frequent, there are other rare and unknown mutations that can create large problems in designing preventive programs. We detected and explained the common mutations in north-western Iran previously and detection of the rare and unknown mutations could be useful in diagnosis and design of future preventive programs. In this study. 5ml peripheral blood from 20 Azari- beta-thalassemia patients whose mutation was not revealed in the previous study was collected and DNA extraction was done by isopropanol and proteinase k method. initially, samples were examined for the rare mutations: Codon6, Codon16, Codon4l/42, Codon36/37, - 88 and Codon22 by ARMS - PCR techniques and then the unknown cases were directly sequenced. According to our results, Codon15[TGG-TGA], Codon16[-C], Codon36/37[-T], lVSII-848[C-A], IVSII-745[C-G], -28[A-C] and Codon25/26[+T] were recognized and added to the spectrom of beta globin gene mutations in Azerbaijan and Iran. Also, we detected four SNP sites: 5'UTR+20[C-T], Codon2 [CAC-CAT] IVSII-16[C-G] and IVSII-666[T-C] in beta-thalassemia genes. Our results could be useful for developing molecular screening plans and help prenatal diagnosis of beta thalassemia in Azerbaijan, Iran and other neighboring countries

[Survey of susceptibility of cockroaches to pyrethroids insecticides in Sari hospital in 1383]

The medical importance of cockroaches is much greater than what has generally been realized. They are known to carry pathogenic viruses and bacteria such as escherichia coli, Staphylococcus aureus and salmonella species including S.typhi and S.typhimurium. As a result, their eradication is very important. One of the best and effective ways of controlling cockoaches is thought to be the chemical method, which is performed by insecticides. It should be stated that the chemical treatment must be combined with enviromental sanitation, otherwise, the results will not be satisfactory. This study was carried out by using four different insecticides like: Permethrin 0.92% Deltamethrin 0.98% and Supermithrin 0.92% to evaluate their susceptibility and resistance. In this study, the cockroaches were collected from Imam Khomeini and Boo Ali Hospitals in Sari and were tested by different cocentrations of above -mentioned insecticides. Susecptibility of cockroaches was performed according to the standards of World Health Organization [W.H.O]. In this study, all the strains of cockroaches were exposed to the concentration of the above-mentioned insecticides through surface contact method. Imam Khomeini strain showed the LT50 of 20/24, 19/30, 19/64 minutes and Boo Ali hospitals 19/87, 17/6, 18/66 minutes. For susceptible strain, 8/89, 8/8, 8/64 minutes were obtained to Permethrin, Deltamethrin and Sumithrin respectively. On the other hand, LT90 for Imam Khomeini strain were 42/91, 41/21, 37/38 minutes and Boo Ali hospitals 42/59, 40/88, 34/05 minutes. For susceptible strain, 17/58, 18/43, 17/28 minutes were obtained to Permethrin, Deltamethrin and Supermithrin respectively. In surface contact, the resistance ratio [RR] to pyrethroids insecticides [Permethrin, Deltamethrin and Supermithrin] of BoAli hospitals strain was 2/23, 2 and 2/15 respectively, while this ratio for Imam Khomeini strain was 2/2, 2/19 and 2/27. Direct use of permethrin, Deltamethrin and supermithrin has developed the resistance of German cockroaches significantly

Etiology and antibacterial resistance of bacterial urinary tract infections in children's medical center, Tehran, Iran

Urinary tract infection [UTI] is a common bacterial illness in children. Knowledge of the antimicrobial resistance patterns of common uropathogens in children according to local epidemiology is essential for providing clinically appropriate, cost effective therapy for UTI. The aim of this study was to determine the distribution of urinary tract infections in a referral hospital, Children's Medical Center, and determination of in vitro susceptibility of these organisms to antimicrobial agents. Of the 1231 bacterial isolates the most frequent isolates were Escherichia coll [38.66%], Klebsiella spp. [22.25%], Coagulase-negative staphylococci [10.1%], Pseudomonas spp. [8.7%], enterococci [8.28%], Enterobacter spp. [4.1%], staphylococcus aureus [3.24%], and proteus mirabilis [2.9%]. Among Enterobacteriaceae, 79.80% of E. coli were amikacin-sensitive. Of Gram-positive cocci, 66.66% of staphylococcus aureus were vancomycin-sensitive. Our data show the original distribution of uropathogens from UTIs in children referred to Children's Medical Center in Tehran and the emergence of multidrug resistant strains