[Evaluation of nucleostemin gene expression as a new molecular marker in breast tumors]

Nucleostemin is one of the stem cell enriched proteins which encodes a novel nucleolar GTP-binding protein found at high levels in the adult and embryonic stem [ES] cells but not in terminally differentiated cells. It is also expressed in tumor cell lines as well as in the several types of human cancers. Due to the increasing rate of breast cancer in recent years, in the present study we evaluate the usefulness of Nucleostemin as a potential diagnostic and therapeutic molecular marker in breast tumors. A total of 41 tumoral and 20 non-tumoral adjacent tissues were studied by Semiquantitative Reverse Transciptase-Polymerase Chain Reaction [RT-PCR]. Beta 2m was used as an internal control. Data were analyzed through SPSS software. According to the obtained results, nucleostemin is a proliferation marker with higher eapression in breast tumors rather than in adjacent normal tissues. Nucleostemin expression level was significantly correlated with profilertion potential of breast benign tumors [p<0.05]. The expression of Nucleostemin was significantly correlated with the advanced stages of breast tumors [p<0.05]. Nucleostemin expression level may be used in estimating tumor size and as a potential prognostic marker for determinig breast tumors stage and future metastases. Moreover, nucleostemin inhibition can be an effective sterategy in decreasing the proliferation of breast tumor cell lines

[Evaluation of the expression of survivin-3b and survivin-3alpha ,the novel splice variants, as diagnostic tumor markers in thyroid cancer]

Survivin is a new member of inhibitor of apoptosis protein family [IAP] that plays an important role in the regulation of cell cycle and inhibition of apoptosis. Distinct expression of this gene in tumoral cells versus normal cells introduces it as the fourth major transcriptome in cancers. Thyroid carcinoma is the most common endocrine malignancy. Considering the highly heterogeneous nature of tumoral and non-tumoral thyroid nodules from the pathological viewpoint and also in regard to the absence of appropriate molecular markers, extensive efforts have been made to find a specific molecular tumor marker for diagnosis of thyroid tumors. Studies have been demonstrated that the expression pattern of survivin and its splice variants was different in cancerous tissues compared to normal tissues. In this study we evaluated expression of survivin-3b and survivin-3alpha, the novel survivin splice variants, as diagnostic markers for thyroid cancer. This was a descriptive study. 77 thyroid specimens; including 49 tumoral, 14 non-tumoral and 14 tumor margin samples were collected and expression of survivin-3b ands-3alpha was investigated by hemi-nested RT-PCR method. Tumoral samples showed the highest expression of survivin-3b and survivin-3alpha and the lowest expression was detected in the specimens of tumor margins. In this study we demonstrated the expression of survivin-3b and survivin-3alpha in thyroid tumors for the first time. In conclusion significant expression of survivin-3b and survivin-3alpha splice variants in tumoral cells shows their roles in thyroid cancer progression and their efficiency as molecular markers for detection and classification of tumoral and nontumoral thyroid nodules

[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

[Evaluating the expression of survivin and its splice variants; 2B and AEx3 as new diagnostic molecular markers in thyroid cancer]

Thyroid cancer is the most common endocrine malignancy. Because of the highly heterogeneous nature of tumoral and non-tumoral thyroid nodules and lack of suitable clinico-pathological criteria and absence of appropriate molecular markers, scientists have been trying to find a molecular tumor marker for specific diagnosis of thyroid tumors. Recent attention has been paid to Survivin, a novel member of the Inhibitor of Apoptosis Protein Family [IAP], as a new molecular marker in cancer. Studies have demonstrated that Survivin and its splice variants have different expression in cancerous tissues compared to normal tissues. In this study the expression of Survivin and its splice variants; 2B and [delta] Ex3 were evaluated as new diagnostic molecular markers in thyroid cancer. Tissue samples were collected from 61 thyroid specimens including 14 tumor margins, 11 non-tumoral and 36 tumoral samples. Expression levels of Surviving and its variants were measured by semi quntitative RT-PCR. Expression level of Survivin in tumor samples was significantly higher compared with surgical margins and non tumural tissues. There was also a significant increase in expression level of Survivin-[delta]Ex3 in tumoral tissues compared with surgical margins. The expression of Survivin 2B in tumors was lower than the non-tumoral tissues. Our data indicated the important role of Survivin in production of thyroid tumors and also revealed that high expression of [delta]Ex3 variant is correlated with nature of thyroid tumors. Therefore, evaluating Survivin gene expression and its recently introduced splice variants may be used in diagnosis and classification of thyroid tumors from non-tumoral lesions

[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

[Evaluation of the effects of cobalt-60 gamma radiation on budding of Triticum aestivum seeds]

Previous studies have reported a significant difference between the effects of low and high dose rate gamma rays. The goal of current study was to determine the dose for enhancing the rate of budding of Triticum aestivum cv Arvand seeds. 5355 seeds with similar phenotype were provided and exposed to gamma rays produced by a CO-60 machine installed in Tabriz Imam Khomeyni Hospital [Theraton-1000, Field Size=10x10cm, SSD=80cm and Dose Rate=155cGy/min]. Then, seeds were exposed in 6 groups [one control group and 5 groups with daily doses of 0, 100, 250, 500 and 750 cGy respectively for 9 days]. 75 seeds from each group were daily counted, sterilized and then transferred to petry dishes. Budding seeds were daily counted and coleoptile length, number of roots and length of roots were measured for each seed in each petry dish after 5 days. Statistical analysis was performed using MSTATC in Random Complete Blocks Design. Our data showed that the optimum dose for maximum budding was 500 cGy per 4 days. Also, we observed that using 500cGy in 4 days was useful for geographical places with short-time growth seasons. Regarding our results, we recommend using 250 cGy per 4 days that is more economical than 500cGy per 4 days. Meanwhile, all examined doses showed a significant decrease in the number of budding seeds after 7[th] day. This can be due to the destructive effects of gamma rays on the budding seeds. We hope our findings help farmers produce crops with high yields