Bi technology IranianJournal of

Background: The High Resolution Melting (HRM) method is a new scanning method for detecting unknown changes in DNA and its advantages have persuaded researchers to recruit it as a screening method. Objectives: Here, we developed a HRM method to screen R188H SNP (rs3218536) of XRCC2 and compared the results with a well known PCR-RFLP technique. Materials and Methods: Genomic DNA samples from 350 healthy individuals were obtained. PCR-HRM analysis and PCR-RFLP method were performed simultaneously. Results: Three different melting profiles corresponding to three different genotypes recognized by HRM analysis. The results of PCR-RFLP showed no discrepancy. Conclusions: We concluded that the HRM technique can be used as a screening method for rapid discrimination of R188H genotypes in XRCC2 gene

Exogenous coenzyme Q10 modulates MMP-2 activity in MCF-7 cell line as a breast cancer cellular model

<p>Abstract</p> <p>Background/Aims</p> <p>Matrix Metalloproteinases 2 is a key molecule in cellular invasion and metastasis. Mitochondrial ROS has been established as a mediator of MMP activity. Coenzyme Q₁₀contributes to intracellular ROS regulation. Coenzyme Q₁₀beneficial effects on cancer are still in controversy but there are indications of Coenzyme Q₁₀complementing effect on tamoxifen receiving breast cancer patients.</p> <p>Methods</p> <p>In this study we aimed to investigate the correlation of the effects of co-incubation of coenzyme Q10 and N-acetyl-L-cysteine (NAC) on intracellular H2O2 content and Matrix Metalloproteinase 2 (MMP-2) activity in MCF-7 cell line.</p> <p>Results and Discussion</p> <p>Our experiment was designed to assess the effect in a time and dose related manner. Gelatin zymography and Flowcytometric measurement of H2O2 by 2'7',-dichlorofluorescin-diacetate probe were employed. The results showed that both coenzyme Q10 and N-acetyl-L-cysteine reduce MMP-2 activity along with the pro-oxidant capacity of the MCF-7 cell in a dose proportionate manner.</p> <p>Conclusions</p> <p>Collectively, the present study highlights the significance of Coenzyme Q₁₀effect on the cell invasion/metastasis effecter molecules.</p

Effect of Progesterone-only Contraception on Vitamin D in Human Milk

Objective: Nowadays progesterone contraceptives are being used by lactating women. In the last century elements effective in bone growth in infants, such as calcium, phosphorus and vitamin D have been evaluated. Vitamin D has been mentioned mainly in relation to rickets prevention in infants. This study was initiated in order to investigate the effect of Progestin-only contraceptives on breast milk vitamin D. Material & Methods: In this cohort study 138 lactating womenwere assigned to either the hormonal (52 participants) or non-hormonal (86 participants) groups according to their chosen method of contraception. The characteristics and goals of the study were explained in detail and informed consent was obtained from all subjects before inclusion in the study. Research units started their method of contraception 45 (7) days postpartum and continued at least for 6 months afterward. 10 ml milk samples were obtained before the baby nursed at 6 mos postpartum, and kept at -20 °C until analysis. Vitamin D levels were measured with RIA method. Findings: Vitamin D levels were found to be 11.2 (7.2) nmol/L in the hormonal and 10.67±6.6 nmol/L in the non-hormonal groups, which was lower than other cultures. There was not a significant difference between the vitamin D levels of the two groups statistically (P>0.05). Conclusion: According the results, the consumption of progesterone-only contraceptives had no effect on the vitamin D levels of mothers' milk, although with regard to low levels of vitamin D in the milk of lactating mothers, it is necessary to enrich foods with vitamin D and for mothers to use vitamin D supplements

RELATIONSHIP OF BREAST MILK LEPTIN WITH MATERNAL AND INFANT ADIPOSITY

AbstractINTRODUCTION: Leptin, the product of the obese gene (ob), is synthesized by adiposetissue and contributes to the regulation of energy homeostasis and food intake. Recently,immunoreactive leptin was reported to be present in human milk. The aim of this studywas to determine any possible link between breast milk leptin concentrations andadiposity in exclusively breast-fed infants.METHODS: 130 healthy, exclusively breast-fed infants beyond neonatal period andtheir mothers were included in the study. Infants whose weight for age was above the75th, and between 25th and 75th percentiles were defined as obese (n=65) and non-obese(n=65), respectively. Anthropometric measur ements of infants and mothers were alsomade and breast milk samples were analyzed for leptin.RESULTS: There was no significant difference between breast milk leptinconcentrations of mothers of obese and non-obese infants. Breast milk leptinconcentrations significantly correlated with mothers' body mass index (r=0.54, P&lt;0.001)and weight (r=0.46, P&lt;0.001). There was no significant correlation between breast milkleptin concentrations and weight of infants.CONCLUSIONS: Leptin concentrations of human milk are not different in the mothers ofobese and non-obese infants. Our findings suggest that milk-borne leptin has nosignificant effect on adiposity during infancy.Key Words: Breast milk, leptin, infant, adiposity

The Effects of Maternal Passive Smoking on Maternal Milk Lipid

Passive smoking was long overlooked by those in the medical and legal professions as being harmful to one's health, but in recent years the negative effect of passive smoking has come to the fore in the media and laws have been changed so that less people are obliged to unwillingly suffer from passive smoking, particularly in the workplace and in indoor settings. To study the effects of environmental tobacco smoking exposure during the breast-feeding period on maternal milk lipids. This cohort study was conducted on 45 mothers environmental tobacco smoking exposure and 40 non-exposed post-partum mothers referred to the Shahid Ayat health center, Tehran, Iran. Socioeconomic conditions and the demographic characteristics of exposed and non-exposed groups were recorded. Milk samples were collected twice--at baseline (5-7 days after delivery) and four months after delivery. The samples were reserved at -20°C until assay. Milk lipids including cholesterol, triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) were evaluated. Dietary intake assessment was performed by means of the 24-hour dietary recall questionnaire both times. Maternal occupation status and education levels were significantly different between the two groups. Lipids profiles of milk were significantly higher 5-7 days after delivery in the non-exposed group and four months after delivery. Dietary intake was not significantly different between the two groups. Maternal environmental tobacco smoking exposure affects milk lipids which are essential for infant growth

Zinc and Copper Concentrations in Human Milk and Infant Formulas

Objective: Available accurate data on the concentrations of copper (Cu) and zinc (Zn) in human milk throughout lactation and infant formulas is important both for formulating nutritional requirements for substances and to provide a base line for the understanding the physiology of their secretion. The objective of this study was to analyze the concentrations of zinc and copper in infant formulas and human milk during prolonged lactation. Levels of these metals were examined in relation to selected parameters such as age, weight, height, education and occupation of mothers. Methods: Thirty mothers referred to the selected clinics in Tehran entered the study. Human milk samples were collected at 2 months postpartum. Zinc and copper concentrations were determined by atomic absorption spectrophotometer. Findings: The mean values of Zn and Cu in human milk were 2.95±0.77mg/L and 0.36±0.11 mg/L. The mean values of Zn and Cu in infant formulas were 3.98±0.25 mg/L and 0.53±0.17mg/L. Conclusion: No significant relationship was found between levels of trace elements in human milk and evaluated parameters such as age, weight, height, education and occupation of mothers. The concentrations of zinc and copper in breast milk were lower than those reported in the literature

Assessment of genetic mutations in the XRCC2 coding region by high resolution melting curve analysis and the risk of differentiated thyroid carcinoma in Iran

Homologous recombination (HR) is the major pathway for repairing double strand breaks (DSBs) in eukaryotes and XRCC2 is an essential component of the HR repair machinery. To evaluate the potential role of mutations in gene repair by HR in individuals susceptible to differentiated thyroid carcinoma (DTC) we used high resolution melting (HRM) analysis, a recently introduced method for detecting mutations, to examine the entire XRCC2 coding region in an Iranian population. HRM analysis was used to screen for mutations in three XRCC2 coding regions in 50 patients and 50 controls. There was no variation in the HRM curves obtained from the analysis of exons 1 and 2 in the case and control groups. In exon 3, an Arg188His polymorphism (rs3218536) was detected as a new melting curve group (OR: 1.46; 95%CI: 0.432-4.969; p = 0.38) compared with the normal melting curve. We also found a new Ser150Arg polymorphism in exon 3 of the control group. These findings suggest that genetic variations in the XRCC2 coding region have no potential effects on susceptibility to DTC. However, further studies with larger populations are required to confirm this conclusion

Enterolactone Reduces Telomerase Activity and The Level of Its Catalytic Subunit in Breast Cancer Cells

Objective There is a positive correlation between higher serum phytoestrogen concentrations and lower risk of breast cancer. The activation of telomerase is crucial for the growth of cancer cells; therefore, the aim of this study was to examine the effects of enterolactone (ENL) and enterodiol (END) on this enzyme. Materials and Methods In this experimental study, we performed the viability assay to determine the effects of different concentrations of ENL and END on cell viability, and the effective concentrations of these two compounds on cell growth. We used western blot analysis to evaluate human telomerase reverse transcriptase catalytic subunit (hTERT) expression and polymerase chain reaction (PCR)-ELISA based on the telomeric repeat amplification protocol (TRAP) assay for telomerase activity. Results Both ENL and END, at 100 μM concentrations, significantly (P<0.05) reduced cell viability. However, only the 100 μM concentration of ENL significantly (P<0.05) decreased hTERT protein levels and telomerase activity. Lower concentrations of ENL did not have any significant effects on telomerase activity and hTERT protein levels. Conclusion High concentration of ENL decreased the viability of MCF-7 breast cancer cells and inhibited the expression and activity of telomerase in these cells. Although END could reduce breast cancer cell viability, it did not have any effect on telomerase expression and activity