[Evaluation of the two variants of p63 [d4TAp63 and delta Np73L] expression as molecular markers in the diagnosis and prognosis of breast cancer]

The TP63 gene, as the oldest homologue of TP53, encodes two main N-terminal variants by different promoters: the trans-activating variant, TAp63 with tumor suppressor activity and the amino terminal truncated variant, delta Np63 with oncogenic activity. As the result of the deletion of exon 4 in each of the variants, two further N-terminal variants have been reported for p63 in the last decade: d4TAp63 and delta Np73L, but the exact function of these variants have not been determined in normal and tumoral cells. In this study we evaluated the expression pattern of p63 variants and usefulness of d4TAp63 and delta Np73L as potential diagnostic molecular markers in breast cancer. In this study 30 tumoral and 20 non tumoral samples of marginal tissues were studied by use of Semi-quantitative Reverse Transcriptase-nested PCR [RT-nPCR] method. SPSS16 software was used for data analysis. In all cases with expression of TAp63 and delta Np63 variants, d4TAp63 and delta Np73L variants were always expressed with their long variants simultaneously. In tumoral and marginal samples delta Np73L mean expression level was significantly higher than the mean expression level of d4TAP63 [P = 0.009 and P=0.008 respectively]. delta Np63 expression levels in tumoral and marginal samples were also higher than those of TAp63, which had a statistically significant difference in tumoral samples [P=0.03], but no significant difference was detected in marginal samples [P = 0.11]. The results indicated dominant negative inhibitory activity of delta Np63 and its potential role as an oncogene in breast cancer. delta Np73L variant compared to d4TAp63, in tumoral and non tumoral breast tissues can act as dominant negative variant. Both variants [delta Np73L and d4TAp63] with similar expression patterns to their respective longer variants [delta Np63 and TAp63] and simultaneous expression with their variants are likely to be involved in strengthening the tumor suppressor activity in normal and tumoral breast cells. On the other hand, according to the expression patterns of delta Np73L and d4TAp63 variants, they cannot be considered as molecular markers in the diagnosis of breast tumors

[Detection of P53 gene exons 5 and 6 mutations among east azerbaijani women with breast cancer]

Breast cancer [BC] is the most common invasive malignancy affecting women worldwide. The tumor-suppressor P53 gene [P53] is frequently mutated in breast tumors. To use P53 as a target for therapy, it is important to accurately assess p53 mutation status in tumor samples. A total of 102 tumor samples were collected from breast cancer patients referred to Tabriz hospitals between the 2007-2009 period. DNA was extracted by Proteinase K- Isopropanol method and then performed amplification and sequencing of P53 from exons 5 and 6. Mutations in the P53 gene were detected in 17.6% of the patients. Including 7 polymorphisms [6.68%] and 11 mutations [10.78%]. Overall, 18.2% of the mutations were found in codons 160 [ATG>AAG] and 163 [ATC>AAG] in exon 5. Also 81.8% of the mutations observed in exon 6: codon 193[CAT>AAT], codon195 [ATC>TTC], codon 195 [ATC>AAC], codon 198[GAA>TAA], codon 220 [TAT>TGT], codon 213 [CGA>CTA], and codon 214 [CAT>CG]. No alteration observed in intron5 and all of polymorphism detected in 13399A>G nucleotide of exon 6. The majority of detected mutations are missense that located on DNA-binding domain of P53. This type of mutation usually leads to the production of a mutant protein with a compromised structure and altered DNA-binding capacity. This is the first report of its kind from the East Azarbaijan region. Our results indicate a rather high frequency of exon 6 mutations in P53 among patients with breast cancer. Furthermore, the mutation pattern appears differs from other regions. However, further studies are needed to determine the role of P53 mutations in breast cancer development

[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 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

[Contamination of drinking water with arsenic and its various health effect in the village of Ghopuz]

Arsenicosis is a serious environmental disease caused by chronic exposure to arsenic- usually from drinking water. Signs and symptoms of chronic arsenic poisoning include hyperkeratosis, hyper- or hypopigmentation, and ulcers. Also, the incidence of cancer is increased in the exposed population. There is some evidence of high arsenic levels in drinking water in the village of Ghopuz, located in Hashtrud District, East Azerbaijan province. We evaluated the genetic and health effects of chronic arsenic exposure in the residents of Ghopuz. In this cross-sectional study we determined the prevalence of hyperkeratosis, hyperpigmentation and hypertension in Ghopuz village. The study involved 101 individuals in Ghopuz and 107 in the adjacent village of Mayan, who were all visited by a trained physician. A total of 46 blood samples were collected for kariotyping. The level of heavy metals in water was determined by the Inductively Coupled Plasma [ICP] method. We detected high arsenic levels in the drinking water at Ghopuz [mean concentration in water = 1.03 mg/L]. There were chromosomal defects in the exposed group. Mean systolic blood pressure at Ghopuz [137mmHg, 95% CI: 132-142] was significantly higher than in Mayan [107, 95% CI: 99.9-114]. Also, mean diastolic blood pressure at Ghopuz [82, 95% CI: 79-85] was significantly higher than in Mayan [71, 95% CI: 66-75]. Hyperkeratosis was 34 times more frequent in the exposed population [OR = 34, P< 0.001]. Also, hyperpigmentation was significantly more frequent in the exposed population [OR = 2.4, P < 0.007]. Water arsenic and nitrate levels at Ghopuz were higher than the maximum permissible levels. The prevalence of skin lesions and hypertension is increased at Ghopuz village due to arsenic exposure. There is also some evidence of chromosomal defects in the exposed group. Affected people need appropriate medical care, and safe drinking water should be provided to reduce arsenic exposure

[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

[Evaluation of the effects of sinusoidal vibrational electroshock [with low dose] in sertoli cells, leydig cells and gonadotropin hypophysis hormones [FSH, LH] and Testis tissue]

In 1337 Yokou Sirleti and Loosened bini cured the patients suffering from Schizophrenia using of electrical shock which showed more efficiency than drug. But Honk gonngian psychologists Found that such treatments have opposite chemical effects on patient's brain. In this research the effects of sinusoidal vibrational [with low dose] on Sertoli cells, Leyding cells, gonadotropin hypophysis Hormones [FSH, LH] and Testis tissue were studied. In this study 50 mature male mice categorized in to ten groups [each group five mouse]. One control and 9 groups were treated by 4, 8, 12 tolerable shocks in zero, 13 and 30 volts [Frequency of 35 kHz]. The duration of each shock was 2 minutes which was repeated every other day for 24 days. the specifications of the electrical sock system imput are 220 volts, 50 Hz and 200 mA and its output was tunable. the results of Sertoli and Leydig cells and hormone levels showed that there is a significant difference in the amount of cells and average blood hormones of LH, FSH between control and main groups [p< 0.05]. Our data suggests that electroshock is effective on higher centers of hyphophysis, therefore causes a direct effect on the metabolic activity of the hyphophysis cells and also causes decrease in metabolism and GnRH production. This task has effects on Hyphophysis and at last decrease amount of Sertoli cell, Leyding cells and FSH, LH gonadotropin hormones

Comparative analysis of first-degree relative's karyotype of mentally-retarded children with chromosome abnormalities and normal children

Background and Purpose: The first chromosomal abnormality was first reported in 1959 by Lujeune and his collogues and since then, studies have suggested the relationship between chromosomal abnormalities and retarded phenotype of the affected children. Nowadays, therefore, detecting chromosomal abnormalities is considered the primary diagnostic tool in congenital abnormalities including mental retardations. This study is, therefore, intended to compare the first-degree relatives' karyotoype of mentally-retarded children with such abnormalities with those of normal children

Materials and Methods: in this case - control study Blood samples were taken from 62 relatives of mentally-retarded children and 22 healthy volunteer subjects. After culturing and staining by Gimsa, the chromosomal expansions of blood lymphocytes were examined by a microscope

Results: It was revealed that 75.8% of the experimental subjects [47 cases] had a kind of chromosomal abnormality including 44.7% structural, 25.5% numerical, and 29.78% mixed abnormalities. Also, abnormal karyotype was observed in 82.54% of parents, and 73.4% of the siblings

Conclusion: The high percentage of abnormal karyotype in parent and siblings of the mentally-retarded subjects, in comparison with the control group, suggest their close relationship with abnormal phenotypes in the affected children. Also, the high percentage of children [73.4%] of [at least one of] parents with abnormalities may indicate the significant role of genetics and heredity in transmitting these defects. The consequence of which may be physical and mental disorders in the affected children