[Construction of a phage M13-based gene carrier and examination of its capacity for transgene delivery and expression in eukaryotic AGS cell line]

Due to some drawbacks associated with gene delivery vehicles including viral and non-viral vectors, scientists have continued their efforts to find an ideal gene delivery vehicle. Bacteriophages have been proposed as an attractive alternative gene delivery vehicle in view of their advantages such as protection of transgene, lack of immunogenicity and stability in different conditions. This study has been conducted with the aim of obtaining a construct based on M13 phage and evaluating its capability for delivering transgene to eukaryotic cells. pCMV-Script EX phagemid was constructed by intracellular excision of Lambda-Zap CMV XR vector bearing GFP gene. Packaging of the construct using helper phage resulted in M13-GFP phage particles which used for transfection of human AGS cell line. Finally internalization of the phage particles into the AGS cell line was evaluated by PCR and florescence microscopy. Examination of the treated cells with florescence microscopy indicated that M13-GFP phage particles were able to internalize and express the transgene in eukaryotic cells, but the efficiency of this trasfer was very low. PCR analysis showed that internalization of the M13-GFP gene vehicle to eukaryotic cells was dose dependent. The results indicated that M13 phage could be an appropriate gene delivery vehicle, because it had trivial tropism for eukaryotic cells. This means that after displaying or coupling of appropriate targeting molecules on the surface of phage particles, transgene can effectively be delivered to the target cells

[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

comparison between zinc sulfate and chlorhexidine gluconate mouthwashes in the prevention of chemotherapy-induced oral mucositis

Patients undergoing high-dose chemotherapy for hematological malignancies are susceptible to development of oral mucositis, and no effective modality has been reported for its prophylaxis and treatment. The aim of this study was to evaluate the effectiveness of zinc mouthwash on chemotherapy-induced oral mucositis lesions. In this double-blind randomized trial, patients under chemotherapy for acute leukemia were divided into two test and control groups of 15 patients each. The groups were homogeneous with respect to medical history, tumor characteristics, and therapeutic details. The test group received 10ml 0.2% zinc sulfate mouthwash, and the control group received 10ml 0.2% chlorhexidine gluconate mouthwash, twice a day for a period of two weeks. Spijkervet scale was used to grade the severity of mucositis at every other week during eight weeks. The severity scores were analyzed with repeated measure ANOVA using SPSS 13.0 computer software. Mean severity scores were generally lower in the test group compared to the controls at all four time intervals evaluated; but only, the differences in weeks of 2 and 3 were statistically significant [P=0.025]. Zinc mouthwash used in conjunction with chemotherapy may reduce the severity of oral mucositis lesions in patients with leukaemia

[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 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

comparative study about effectiveness of an exercise therapy program on improvement of gait and balance in left and right hemiplegia

Impairment in the function of the brain caused by deficiency in the brain circulation is called stroke. Stroke is the most important factor of disability in adults. One of the most prevalent consequences of stroke is disability of walking or so-called [gait disorder] so that more than 50 percent of the patients who are in the acute phase of the disease are not capable of walking, and need to a rehabilitation period to reach for walking. In order to compare the effect of exercise therapy on the function of walking and balance and Improvement of depression in left and right hemiplegics patients, a Non-randomized clinical study was done. This study consisted of 13 right- hemiplegics patients with an average age of [60.46 +/- 1.47] and 13 left hemiplegics patients with an average age of [61.92 +/- 2.01] who were matched in age, height and weight. To measure the function of walking the Gait Dynamic Index, function of balance the Berg Balance Scale and Depression Beck Depression Questionnaire were used. The data were analyzed by SPSS software. To compare the effect of exercise therapy on the improvement of walking and balance before and after the treatment in each group t-paired -test and for more comparison of the two groups an independent T-test was used. With analyzing the received data the following results were derived: From the viewpoint of walking, a significant difference in the right hemiplegics group [p<0.001] and the left hemiplegics one [P=0.0] before and after the treatment is derived. From the viewpoint of balance, a significant difference in the right hemiplegics group [p<0.001] and the left hemiplegics group [P=0.0] before and after the treatment was noticed. About the improvement of depression, a significant difference in the right hemiplegics group [p<0.001] and the left one [p<0.00l] before and after the treatment was noticed. There was no significant difference between two groups before [p=0.85] and after [0.36] the treatment from the point of view of walking. There was no significant difference between two groups before [p=0.056] and after [p=0.067] the treatment in balance. There was no significant difference between two groups before [p=0.067] and after [p=0.054] treatment from the viewpoint of depression. This study showed that performing the exercise therapy protocol improves the function of walking, balance and depression in herniplegic patients; never the less, no differences were observed between the two right and left hemiplegics groups

[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