Effects of 35 Hz, 2mT magnetic field on peripheral blood lymphocytes of human in vitro and rat in vivo

In this research we have studied the effects of extremely low frequency magnetic field on the peripheral blood lymphocytes chromosomes of human in vitro and rat in vivo. By this means, we used 60 blood samples of 10 men as well as 40 rats. Of 60 human blood samples, 40 samples (In 2 group of 20) were selected as test groups and placed into the 2mT and 35Hz magnetic field. The blood samples, then, with 20 samples of control groups cultured for chromosomal analysis of lymphocytes cells. Of 40 rats, 30 rats, in 2 groups of 15, were also placed into the same magnetic field, and cultured with 10 control rats for chromosomal analysis of lymphocytes cells. The analysis results of 5373 metaphasic cells from human bloods (3573 test cells and 1800 control cells) showed that there were no significant differences among the percentages of chromosomal aberrations of control (0.16%), 30 and 60 minutes test (0.224%, 0.296%) groups respectively (P30>0.28, P60>0.15). There were no chromosomal aberrations in 3089 metaphasic lymphocytes of rats (2389 test cells and 700 control cells). Keywords: Magnetic field, Extremely low frequency field, Lymphocytes, Chromosom

Evaluation of Mutations in Exons 7 and 8 of TP53 Gene in Breast Cancer Patients from Azarbaijan

BACKGROUND AND OBJECTIVE: Breast cancer is the most common type of malignancy in women, and TP53 tumor-suppressor gene is one of the most commonly transformed genes in human cancers. Accurate assessment of TP53 gene mutations in cancer patients can play an important role in diagnosis, prognosis, or treatment. This study aimed to identify mutations of this gene in breast cancer patients. METHODS: In this descriptive study, 102 tumor samples were obtained from female breast cancer patients from Azarbaijan, Iran. All the participants were referred to hospitals of Tabriz during 2007-2009, and their DNA was extracted by Proteinase K. TP53 gene mutations in exons 7 and 8 and intron 7 were investigated using polymerase chain reaction technique and direct sequencing. FINDINGS: Seven (6.86%) cases of mutation and 14 (13.72%) cases of polymorphisms were identified. Mutations (CGG → CAG) at codon 248 (in two cases) and (CTG → CCG) at codon 257 in exon 7 and G>T mutation in the first nucleotide of intron 7 were observed. In exon 8, GTG>ATG mutation at codon 272, CCT>TCT mutation at codon 278, and three nucleotide deletion at codon 262 were identified. CONCLUSION: The results of this study showed a different pattern of TP53 gene mutation in female breast cancer patients. Further studies could specify the role of TP53 mutations in the progression of breast cance

Evaluation of Nucleostemin Gene Expression as a New Molecular Marker in Breast Tumors

Abstract: Background & Aims: 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. Methods: A total of 41 tumoral and 20 non-tumoral adjacent tissues were studied by Semiquantitative Reverse Transciptase-Polymerase Chain Reaction (RT-PCR). β2m was used as an internal control. Data were analyzed through SPSS software. Results: 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). Conclusion: 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. Keywords: Nucleostemin, Breast neoplasms, Reverse Transcriptace Polimeras Chain Reaction (RT-PCR

Clinicopathological significance of plasminogen activator inhibitor-1 gene polymorphism in breast cancer patients from North West of Iran

Introduction: A common polymorphism 4G/5G in the promoter region of the Plasminogen activator inhibitor-1 (PAI-1) gene has been reported to influence the expression levels of PAI-1. According to the evidence, progression of breast cancer can be associated with elevated levels of PAI-1, it seems that evaluation of a possible correlation between the polymorphism and clinical status of breast cancer patients is reasonable. Methods: This descriptive-analytical study included 160 unrelated patients from North West of Iran. According to established clinical criteria, these paitients were diagnosed with breast cancer. Based on previous study, PAI-1 4G/5G had been determined. In order to investigate the association of this polymorphism with clinicopathological features Fisher&rsquo;s exact tests and SPSS software was used with a significance level of 0.05. Results: All declared features of breast cancer regarding PAI-1 4G/5G polymorphism were investigated. Results indicated that PAI-1 4G/5G polymorphism positive correlation with several traditional prognostic factors, including tumor size, lymph node metastases and tumor stage. Conclusion: Data showed that the patients with 5G/5G genotype are more susceptible to the development of breast cancer, while the paitients with 4G/4G and 4G/5G genotypes show lower sensitivity to the breast cancer. Therefore, the 4G allele likely has a protective role against the development of breast cancer in this cohort

The effect of Teucrium Orientale on Oxidative Stress marker and Liver Function Enzymes in Streptozotocin-Induced Diabetic Rats

Abstract Background & aim: Diabetes is a metabolic disorder characterized by hyperglycemia resulting from perturbation in insulin secretion, action or both. The aim of this study was to investigate the effects of methanol extract of Teucrium orientale on blood glucose and liver damage markers in streptozotocin diabetic rats. Methods: In this experimental research 32 male albino Wistar rats, with body weights of 200 – 240 g were randomly allocated into four groups with 8 rats per each. Control group (normal rats), diabetic rats (received STZ in single dose 60 mg/kg bw, intraperitoneal way and without receive extract), treated normal rats (received T. orientale 200 mg/kg bw, oral gavage) diabetic treated rats (received STZ in single dose 60 mg/kg bw, intraperitoneal way and received T. orientale 200 mg/kg bw, oral gavage). Afterwards 21 days, blood glucose, malondialdehyde (MDA) in liver and the activity of the liver enzymes (ALT ،AST،ALP( were evaluated. The collected data were analyzed using SPSS software via the one-way ANOVA. Resulths: This study demonstrated that sSerum levels of glucose in T. orientale treated diabetic group (222 ± 9.8) were significantly lowerdecreased than in comparison with diabetic ratsgroup (572 ± 8) (P < 0.01).The MDA level in T. orientale treated diabetic group (1.01 ± 0.04) were significantly decreased in comparison with diabetic rats (1.25± 0.54) (P < 0.05). The ALT, AST and ALP levels in T. orientale treated diabetic group were also significantly decreased compared with diabetic rats (P < 0.01). Conclusion: Methanol extract of T. orientale had antidiabetic effects and consequently might alleviate the liver damage caused by streptozotocin-induced diabetes mellitus. Keywords: Diabetes, Teucrium orientale, ALT, AST, ALPLiver enzymes, streptozotoci

Calprotectin (S100A8/S100A9)-induced cytotoxicity and apoptosis in human gastric cancer AGS cells: Alteration in expression levels of Bax, Bcl-2, and ERK2

Calprotectin is a heterodimeric EF-hand Ca2+ binding protein that is typically released by infiltrating polymorphonuclear leukocytes and macrophages. This protein is a key player linking inflammation and cancer. Due to the increased levels of calprotectin in different inflammatory diseases and cancer, it is considered as a marker for diagnostic purposes. In this study, we evaluated the mechanism of cell viability and apoptotic-inducing effects of recombinant human calprotectin (rhS100A8/S100A9) on the gastric adenocarcinoma (AGS), the most common type of gastric cancer cell line. AGS cells were exposed to the different concentrations (5–100 μg/ml) of calprotectin for 24, 48, and 72 h, and cell viability was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptotic-inducing effects of calprotectin were evaluated by sub-G1 cell cycle assay and Annexin V/propidium iodide double staining. Furthermore, real-time polymerase chain reaction and Western blot analysis were performed to evaluate the mechanism of action of calprotectin. Our findings indicated that calprotectin inhibits growth and viability of AGS cells in a time- and dose-dependent manner. The half-maximal inhibitory concentration values were measured as 85.77, 79.14, and 65.39 μg/ml for 24, 48, and 72 h, respectively. Additionally, we found that calprotectin downregulated the expression of antiapoptotic protein Bcl-2 and upregulated proapoptotic protein Bax in a time- and concentration-dependent fashion. Calprotectin also slightly upregulated the expression of extracellular signal-regulated protein kinase 2 (ERK2), while it significantly decreased the levels of phospho-ERK in a timedependent manner. Overall, these findings indicated that calprotectin has cytotoxicity and apoptosis-inducing effects on AGS cell lines in high concentration by modulating Bax/Bcl-2 expression ratio accompanied by inhibition of ERK activation

Study of the Expression of Survivin & Its Splice Variants; ΔEx3, 2b and 3b as Diagnostic Molecular Markers in Breast Cancer

Introduction: Survivin is a new member of the Inhibitor Apotosis Protein family (IAP) which plays an important role in the regulation of both cell cycle and apoptosis. Its distinct expression in tumor cells as compared to normal adult cells introduces Survivin as the fourth transcriptom demonstrated in tumors. Breast cancer is the most common malignancy among women and scientist`s efforts to classify it has lead to various molecular subtypes and controversial results. Because of the high prevalence of these tumors and lack of suitable molecular markers for diagnosis and prognosis, there are ongoing efforts to find molecular markers which can distinguish nontumoral from tumor tissues. In this study we evaluate the potential usefulness of Survivin and its splice variants ΔEx3, 2b and 3b as molecular markers in breast cancer. Methods: We studied 18 tumor and 17 non tumor adjacent tissues. Transcription levels were measured by Semiquantitative Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and normalized by ß2m as an internal control. Results: 1)Survivin and its splice variants; Δex3, 2b and 3b showed differentially higher expression levels in tumors than adjacent normal tissues. 2) The expression levels of Survivin, Survivin-ΔEx3 and Survivin-3b were significantly correlated with the type of tumors. 3) Survivin-2b was expressed in a few samples. 4) Survivin-3b was detected only in tumor samples. Also, our results showed that ΔEx3 variant can be introduced as a dominant expressed variant in breast cancer. Conclusion: Our data indicated that the expression of Survivin, Survivin ∆Ex3 and especially, Survivin-3b were correlated with cancerous nature of tumors and Survivin-∆Ex3 was the most common expressed variant in breast carcinomas. These results besides confirming the potential usefulness of Survivin and its splice variants as molecular markers in breast cancer, demonstrated the role of the gene and its splice variants, especially 3b in these lesions that enables distinguishing normal from tumor tissues. Therefore, evaluation of the expression of Survivin and its splice variants might be used as markers to stratify breast cancer patients for more optimal treatment modalities or it could be a promising new target for therapy

Investigating the KLF4 Gene Expression as a New Molecular Marker in Breast Tumors

Introduction: Krupple-like factor4 is a transcription factor which is involved in many cancers including Breast cancer. Breast cancer is the most common malignancy among women. Due to the high prevalence of these tumors, there are ongoing efforts to find molecular markers which can recognize nontumoral from tumoral tissues. Therefore, the aim of this study was to evaluate the potential usefulness of KLF4 as a potential diagnostic and therapeutic molecular marker in breast cancer. Methods: In the current study, 31 tumoral and 21 non tumoral adjacent tissues were evaluated. Transcription levels were measured by Semiquantitative Reverse Transciptase-Polymerase Chain Reaction and were normalized by the ß2m as an endogenous PCR control. Data was analyzed by spss software, one-way ANOVA and T-test. Results: The results showed that: 1) KLF4 is over expressed in Breast tumors rather than adjacent normal tissues. 2) KLF4 is an oncogene in breast tumors (at least in IDC type). 3) The KLF4 expression levels are related significantly with nature of malignant breast tumors. Conclusion: Findings do not confirm KLF4 as a diagnostic marker in classification and identification of tumoral tissues from non-tumoral ones in breast, but we can use this marker to identify at least 50% of invasive Ductal Carcinoma in breast and utilize it as a potential predictive factor to demonstrate severity degree in various tumors

Comparison between the Plasma Levels of Long Noncoding RNA BDNF-AS in Patients with Alzheimer\'s disease and Healthy Subjects

BACKGROUND AND OBJECTIVE: Diagnosis of Alzheimer's disease usually occurs when serious damages have occurred in the brain and common treatments are ineffective in preventing it. One of the RNAs involved in Alzheimer's disease is a long noncoding RNA, called BDNF antisense (BDNF-AS). The aim of this study is to determine the presence and compare the BDNF-AS levels in plasma of Alzheimer's patients and healthy subjects, and to evaluate its potential as a plasma marker for Alzheimer's disease. METHODS: In this case-control study, 30 patients with late-stage Alzheimer's disease and 30 healthy subjects without neurological disease who matched the patients in terms of age were selected by a specialist according to the criteria for clinical diagnosis of Alzheimer's disease and their intravenous blood samples were collected. The plasma of the blood samples was isolated and total plasma RNA was extracted. After cDNA synthesis, the presence of BDNF-AS in plasma was examined by PCR. Finally, the relative level of BDNF-AS transcripts in plasma samples of patients with Alzheimer's disease and healthy subjects was evaluated using Real Time PCR. FINDINGS: The results of this study showed that long noncoding RNA BDNF-AS was present in the plasma of patients and controls. Comparison of Real Time PCR data showed that BDNF-AS levels in the plasma of patients (0.107±0.021) showed significant increase compared to healthy subjects (0.039 ± 0.006). CONCLUSION: The results of this preliminary study indicate that the levels of long noncoding RNA BDNF-AS in plasma can be used as a blood/plasma marker for the diagnosis of Alzheimer's disease