Systemic chemotherapy induces microsatellite instability in the peripheral blood mononuclear cells of breast cancer patients

INTRODUCTION: Systemic chemotherapy is an important part of treatment for breast cancer. We conducted the present study to evaluate whether systemic chemotherapy could produce microsatellite instability (MSI) in the peripheral blood mononuclear cell fraction of breast cancer patients. METHODS: We studied 119 sequential blood samples from 30 previously untreated breast cancer patients before, during and after chemotherapy. For comparison, we also evaluated 20 women who had no relevant medical history (control group). RESULTS: In 27 out of 30 patients we observed MSI in at least one sample, and six patients had loss of heterozygosity. We found a significant correlation between the number of MSI events per sample and chemotherapy with alkylating agents (P < 0.0001). We also observed an inverse correlation between the percentage of cells positive for hMSH2 and the number of MSI events per sample (P = 0.00019) and use of alkylating agents (P = 0.019). CONCLUSION: We conclude that systemic chemotherapy may induce MSI and loss of heterozygosity in peripheral blood mononuclear cells from breast cancer patients receiving alkylating agents, possibly mediated by a chemotherapy-induced decrease in the expression of hMSH2. These effects may be related to the generation of secondary leukaemia in some patients, and may also intensify the genetic instability of tumours and increase resistance to treatment

Functional Characterization of FLT3 Receptor Signaling Deregulation in Acute Myeloid Leukemia by Single Cell Network Profiling (SCNP)

Molecular characterization of the FMS-like tyrosine kinase 3 receptor (FLT3) in cytogenetically normal acute myeloid leukemia (AML) has recently been incorporated into clinical guidelines based on correlations between FLT3 internal tandem duplications (FLT3-ITD) and decreased disease-free and overall survival. These mutations result in constitutive activation of FLT3, and FLT3 inhibitors are currently undergoing trials in AML patients selected on FLT3 molecular status. However, the transient and partial responses observed suggest that FLT3 mutational status alone does not provide complete information on FLT3 biological activity at the individual patient level. Examination of variation in cellular responsiveness to signaling modulation may be more informative.Using single cell network profiling (SCNP), cells were treated with extracellular modulators and their functional responses were quantified by multiparametric flow cytometry. Intracellular signaling responses were compared between healthy bone marrow myeloblasts (BMMb) and AML leukemic blasts characterized as FLT3 wild type (FLT3-WT) or FLT3-ITD. Compared to healthy BMMb, FLT3-WT leukemic blasts demonstrated a wide range of signaling responses to FLT3 ligand (FLT3L), including elevated and sustained PI3K and Ras/Raf/Erk signaling. Distinct signaling and apoptosis profiles were observed in FLT3-WT and FLT3-ITD AML samples, with more uniform signaling observed in FLT3-ITD AML samples. Specifically, increased basal p-Stat5 levels, decreased FLT3L induced activation of the PI3K and Ras/Raf/Erk pathways, decreased IL-27 induced activation of the Jak/Stat pathway, and heightened apoptotic responses to agents inducing DNA damage were observed in FLT3-ITD AML samples. Preliminary analysis correlating these findings with clinical outcomes suggests that classification of patient samples based on signaling profiles may more accurately reflect FLT3 signaling deregulation and provide additional information for disease characterization and management.These studies show the feasibility of SCNP to assess modulated intracellular signaling pathways and characterize the biology of individual AML samples in the context of genetic alterations

The Effects of Coping Therapy on General Health of Pregnant Women with High Risk of Genetics Abnormalities in their Fetus

Introduction: The physiological changes in pregnancy lead to the psychological pressures. Therefore, there is a need for applying cognitive behavioral and emotional confronting to encounter these problems. The present research investigated the effect of coping therapy on general health of pregnant women with high risk of genetic abnormalities in their fetus. Methods: The present study was a semi experimental research. Pre and post tests were used to investigate coping therapy between 30 pregnant women who were referred to Khatomolanbia Genetic Clinic, Yazd, Iran. All the women had pregnancy screening test with high risk of genetics abnormalities in their fetus. They were divided randomly into two groups of case and controls. The test of GHQ was performed in both groups, then the case groups went under 8 sessions of teaching coping therapy each lasting 120 min. After finishing the sessions, post test was performed and analyzing the data using descriptive statistical index and covariance analysis test. Results: Teaching coping therapy to case group caused improvement in their GHQ mark, and this change was significantly different from the change in the GHQ mark of control group. In addition, there was a significant decrease in anxiety, depression and physical signs and an improvement of social function in case group compare to the control group. Discussion: Teaching coping therapy can improve the general health of pregnant women with high risk of genetic abnormalities in their fetus. Therefore, presenting educational courses to the women can improve their general health indices in addition to preventing the probable effects of stress on fetus

Examination and Comparison of Various Methods of DNA Extraction from Samples of Blood, Bone Marrow Slides and Peripheral Blood

Introduction: DNA extraction methods are very important for genetics research and diagnostic tests. In These methods are also important for detecting genetic diseases or in cancers for investigating genetic changes during cancer progression or treatment. Therefore, selection of the best method for DNA extraction from different samples such as bone marrow (BM) or peripheral blood (PB) and their slides is very important. Methods: In the present research, DNA was extracted from 5 different samples including; 1-PB, 2-BM slides stained by Gimsa method, 3-Gimsa stained PB slides from archives, 4-new Gimsa stained PB slides and 5-non stained new PB slides by 3 different methods; salting-out, boiling and phonal chloroform method. In all of the groups, three DNA parameters were investigated; 1-OD (Optimal Density), 2-DNA concentration and 3-Outcome (PCR results). Results: The best DNA quality was achieved by salting-out method (OD=1.74), while the worst quality was by the boiling method (OD<1.0). The DNA quality of all the samples was similar in the salting-out and phenol chloroform methods. Regarding DNA quantity, the best result was from boiling method (6.7µg / ml). The least amount of DNA was obtained by the phenol chloroform method and salting-out method also resulted in the least quantity of extracted DNA. Regarding the outcome of DNA extraction or the PCR results, all the three methods showed 100% positive results for peripheral blood samples, while boiling method had the best outcome for BM slides, archive stained PB slides, new stained PB slides and non-stained PB slides (100%, 88%, 84% and 72% respectively). Discussion: The present research indicated that except non stained PB slides, the DNA extraction from all other samples showed very good results. In addition, the research showed that there is no difference in DNA extraction of new or archive slides

Investigating Frequency of GSTT1 and GSTM1 Genes Null Genotype in Men with Varicocele and Its Association with the Sperm Parameters

Introduction: GSTM and GSTT are subclasses of glutathione s-transferase that is present on human sperm surface and plays an important role against oxidative stress. This study aimed to investigate the polymorphisms of GSTT1 and GSTM1 in regard to sperm parameters. Methods: This case-control study involved 46 men with varicocele and 48 men without varicocele. Semen analyses were carried out according to WHO guidelines. Blood DNA was extracted using salting out procedures. Polymorphism of GSTT1 and GSTM1 genes was determined trough multiplex-PCR respectively. Results: Frequencies of GSTM1 null genotype in men with varicocelc and men without varicocele groups were 60.9 and 41.7 respectively. There were no statistically significant differences between Gstm1 null and positive genotype in two groups (p>0.05). Frequencies of gstt1 null genotype in case and control groups were 47.8 and 50 respectively. There were no statistically significant relationship between gstt1 null and positive genotype in two groups (p>0.05). Conclusions: Deficiency of enzyme activity in gstm1 null genotype did not affect morphology as well as slow and quick progressive of sperm but caused the significant decrease in count of sperm between gstm1 null and positive genotype. In the case of gstt1, gstt1 null genotype did not affect sperm parameters that may be related to compensate activity of other genes in this super family

Inhibitory Effect of Eicosapentaenoic Acid (EPA) on sFlt-1 (Soluble VEGF Receptor-1) Expression in Trophoblast Tumor Cell Line JEG-3

Introduction: Previous studies have shown altered levels of n-3LCPUFA in the pathophysiologcal conditions such as preeclampsia .Also elevated expression of sFlt-1 in preeclampsia plays a major role in the pathogenesis of this serious disorder especially in reduced placental oxygenation. The present study examines the hypothesis that Eicosapentaenoic acid (EPA; 20:5), an omega-3 long-chain polyunsaturated fatty acids (n-3LCPUFAs), may attenuate sFlt-1 gene and protein expression in JEG-3 cells treated with induced hypoxia-like conditions by (DMOG) -induced hypoxia-like conditions. Methods: JEG-3 cells were pretreated with DMOG incubated with EPA. Protein expression of sFlt-1 was measured by enzyme-linked immunosorbent assay (ELISA). Messenger RNA expressions of sFlt-1 was determined by and RT Real Time-PCR. Results: Our results showed that incubation of JEG-3 cells with DMOG cause a significant elevation in mRNA levels and protein secretion of sFlt-1(P < 0.05). In contrast, EPA decreased the mRNA expression and protein secretion of sFlt-1(P < 0.05). Also mRNA expression and protein secretion of sFlt-1 inhibited cells treated by both EPA and DMOG (P=0.261, P =0.077 respectively). Conclusion: These findings confirm previous studies that hypoxia caused elevation in sFlt-1 gene expression and protein secretion. Also our studies reveal that effects of n-3 fatty acids in restraining preeclampsia complications may be mediated by suppressing the gene expression and protein secretion of sFlt-1 under hypoxia conditions. This data provide evidence that n-3 LCPUFA can exert its effects through inhibition of the HIF pathwa

Bunium Persicum(Black Caraway) of Yazd province: Chemical assessment and Evaluation of its Antioxidant Effects

Introduction: Medicinal plants are valuable natural resources that are nowadays known as safe pre-materials for drug industries in the advanced countries. Black caraway is one of the most important and economical medicinal plants that can be grown naturally in some regions of Yazd province. The essence of the plant are used in treatment of diarrhea, dyspepsia, fever, flatulence, stomachache, hemorrhoids, hiccoughs, as an antihistaminic and more numerous uses in medicine industries. This plant shows several therapeutic effects on digestive and urinary tract disorders and is well known as an anti-convulsant, anthelmintic, anti-asthma and antidyspnic drug in Iranian traditional medicine. Bunium Persicum oil is capable of suppressing the initial stage of an inflammatory process. Also, this plant is used for culinary purposes and for flavoring foods and beverages. Methods: In this research, the fractionization of seed essence was done by GC method and the compounds were recognized. The DPPH test was used for estimating anti oxidant effects and the Follin-Ciocalteu method was used for estimating quantity of phenol compounds. Results: The results revealed that- γTerpinene had the highest percentage in the essence of the native black caraway of Yazd province. Also, the antioxidant test showed the high antioxidant effect of the native black caraway of Yazd province with an IC50 of 2.85 µg.mg-1 and the phenolic component percentage of 117.09mg.g1. Conclusion: This result was much higher than previous similar studies about black caraway

New Approach for the Synthesis, Entrapment of Hydrophilic Drugs and Evaluation of Physico-Chemical Characteristics of PH-sensitive Nano-Liposome: Improving Therapeutic Efficacy of Doxorubicin in Order to Treatment Bone Tumor and Reducing the Side Effects of Doxorubicin

Introduction: Cancer is one of the most harmful disease throughout the world. Doxorubicin is an anti-cancer agent, used in the treatment of various types of the cancer such as bone cancer. There are several adverse effects related to clinical usage of Doxorubicin for long time. The present study aimed to investigate the reducing side effects and enhancing the therapeutic effect by liposomal carrier. Methods: Liposomes containing DPPG and cholesterol with the molar ratio of 70:30 with the Doxorubicin were synthesized by pH- gradient method. The average diameter of nanoparticles and surface charge was determined by Zeta-Sizer instrument. The amount of drug loaded and drug-released was determined using dialysis. The surface morphology and internal lamella was evaluated by TEM and SEM. Results: The average size of liposomal Doxorubicin obtained using Zeta-Sizer was 126 nm. The encapsulation efficacy of liposomal Doxorubicin was 89%. The total amount of drug release during 48 hours in acidic medium studied by dialysis technique was 46%. Conclusion: In this study, investigation of loading Doxorubicin into nano-liposome with the slow- released kinetic was carried out to improve the solubility and bioavailability of Doxorubicin in order to delivery to osteosarcoma cell line