Alpha deposit into blood: A new method to evaluate infertility of women by measuring the level of LH, FSH and HCG
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1 Alpha deposit into blood: A new method to evaluate infertility of women by measuring the level of LH, FSH and HCG Najeba F. Salih 1 *, Mohamad S. Jaafar 1 1. Medical Physics and Radiation Science Research Group, School of Physics, Universiti Sains Malaysia, USM, Penang, Malaysia. Abstract This study examines the effect of alpha particles radioactive exposure on pregnancy hormones such as luteinizing hormones (LH), follicle stimulating hormone (FSH), hormone chorionic gonadotropin (HCG), progesterone and estradiol, in order to investigate the prevalence of infertility. Done Hormonal analysis of the women blood samples and irradiated by radium source after then carried out the hormone testing. This disparity supports the hypothesis that the magnitude of changes in pregnancy hormones concentration is indicative of the effect of radiation on the human tissues. To ascertain the level of ovarian reserve, the FSH level was measured, since it correlates inversely with the number of eggs present in ovaries. High levels of FSH were observed in women with low ovarian reserve. Significant differences were found before and after radiation. For LH (p<0.001), HCG (p<0.001), and estradiol (p<0.001), the levels of these hormones significantly reduced after radiation, while elevation observed for progesterone (p<0.003), FSH (p<0.003). This study concluded that FSH is significantly increased by irradiation which subsequently leads to impaired fertility. Keyword: FSH, LH, pregnancy hormones concentration, Estradiol, women infertility, alpha particles 1- Introduction Infertility is defined as the inability to conceive after 1 year of unprotected intercourse (1). Infertility has become a significant problem in society, affecting 15% of reproductive-aged couples. Many cases of infertility appear to be the result of defects in germ cell development that led to reduced numbers or quality of gamete (2, 3). The ovaries produce hormones like Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) which control reproductive functions in the body (4). The deficiencies of these essential pregnancy hormones can result in the absence of menstruation and infertility in women (5). On the other hand, radiation is a phenomenon that increases risk of early puberty, alters menstruation, decreases fertility, and increases miscarriage rates and early menopause occurrences (6). After exposure to radiation, one-third of the deposited decay products from radon are transported * Corresponding Address: Medical Physics and Radiation Science Research Group, School of Physics, Universiti Sains Malaysia, USM, Penang, Malaysia. najebafarhad@yahoo.com, Tel: from the lungs into the blood or absorbed into the bloodstream through the stomach or intestinal walls, and then distributed throughout the body. The radon is capable of inducing significant chromosome damage at very low doses and dose-rates (7). Radiation cause pathological changes in chromosomes which ultimately results in conversion of normal cells to cancer cells. When alpha particles enter a cell, ionization can damage critical molecules or structures in the cell directly by hitting a particularly sensitive atom or molecule in the cell. The damage from this is irreparable, and the cell either dies or malfunctions. The effect of the ionization of incident alpha particles depends on its energy and time of exposure, and can cause an excitation of molecules in blood which is able to reach all body tissues, permeate through the blood circulation system and gonads, resulting in changes to hormone parameters that cause the weakness in fertility of women (8). Ovulation is the process of releasing a mature egg from the ovary for fertilization or pregnancy which a woman s body goes through for about every 24 to 32 days if she is not pregnant (9). There are a lot of hormones that 21
2 affects ovulation, and they include, shown in Figure 1(1). 1. Luteinizing hormones (LH): LH is found in both males and females, and is essential for reproduction. In females, LH levels are normally low during childhood and at the time of menstruation and become high after menopause. 2. Follicle stimulating hormone (FSH). This is the main hormone involved in producing mature eggs in the ovaries. Both high and low levels can reduce fertility. FSH can be measured easily from blood tests. The amount produced is an indicator of the number of eggs in the ovaries (10). 3. Hormone chorionic gonadotropin (HCG): This is similar to LH and causes the empty follicle to continuously produce progesterone, the hormone needed for pregnancy. Pregnancy tests usually measure the level of HCG because its presence indicates that a woman is likely pregnant. HCG is also the first hormonal factor to be present at high concentrations at the foetal maternal interface. During this time, Progesterone (P) acts in multiple ways to maintain pregnancy via actions on the000 endometrium (1). 4. Progesterone: This is the hormone that prepares the uterus for pregnancy. When an egg is fertilized, it sends out a hormone (HCG) to let the body know that it s present(11). The blood sample is taken on the 21st day of a regular 28 day cycle (12). The primary action of progesterone is to maintain pregnancy. This hormone facilitates the LH surge, transforms the endometrium from a proliferative to a subordinal state, together with estradiol (13). Hormone testing of progesterone is helpful in the investigation of infertility. A progesterone concentration above nmol/l confirms ovulation occurred in that cycle. Lower values indicate disruptive ovulation (8). Estradiol: These are hormones produced by the ovaries. They control body functions, and are sometimes referred to as female hormones because women need higher amounts than men do for normal health. Recently, endocrine assays have been used in epidemiologic studies to monitor ovulation and early pregnancy loss in populations exposed to environmental hazards. Measurements of HCG in blood and urine samples have been used to confirm pregnancies (14) Figure 1: The relation between hormones of pregnancy and ovulation 2. Hormone tests Hormone testing is important in the investigation of infertility. Laboratory blood studies are done to measure certain hormones that play a role in fertility. Tests on luteinizing hormone (LH) and folliclestimulating hormone (FSH) are carried out at the start of the menstrual cycle. Measurement of blood progesterone level is performed at day 21 of the menstrual cycle to confirm that ovulation has 0000 occurred. Other hormones measured include estradiol. These tests were conducted to assess the various hormone levels that contribute to the reproductive process. The objective of this study is to investigate the effect of alpha particles on the pregnancy hormones such as LH, FSH, HCG, progesterone, estrogenal in the blood of women that had infertility problem. 22
3 3. Materials and Method Hormonal analysis and in vitro study of the human blood samples were conceptualized to explain the effects of radiation on pregnancy hormones (FSH, LH, HCG, Progesterone, and Estradiol) via the evaluation of alpha particle density. The following steps were taken during the tests: 1. Blood samples (V = 3ml) were aseptically obtained and analysed for 11 women from Iraq Kurdistan region who have decreased fertility and infertility. The age of women ranged from 20 years to 42 years. Blood samples were collected from each subject using disposable syringe, and immediately transferred into a tube. The tests were further conducted in PROLAB - Pulau Pinang, Malaysia to analyze fertility hormones; Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) Progesterone (P), Estradiol (E) and Human chorionic gonadotropin (HCG), using 2 automated machines (Mini Vadis, Machine model). The first one was ELISA Plate Analyzer (INDIA PVT. LTD) while the other was ROBONIK W2002 ELISA Plate Washer and Analyzer (INDIA PVT.LTD). This analytical procedure was the control test for the blood samples, which preceded the irradiation of the samples, 2- The irradiation of samples was then carried out in the biophysics Lab for 30 minutes using 226Ra (5µCi) radon source. All blood samples were exposed to the source at the same distance. After irradiation, the samples were taken back to hormone analysis Lab (PROLAB) for a repeat of all the tests, and the results were afterwards displayed as report number two. The automated hormone analyzer machine was used to measure chemical properties of irradiated and non irradiated blood samples. This machine has many advantages which include; speed, simplified usage and its ability to read irradiated and non-irradiated blood samples or otherwise solely if necessary, therefore easier to test a large number of samples. 4. Statistical analysis SPSS for data All statistical calculations were performed using SPSS for Windows, Standard version The hormone and blood data were analyzed for irradiation. Paired t test was used for LH, HCG, progesterone and blood ph because their results were parametric values. Inversely observed for FSH, Estradiol and urine ph where Wicoxon Signed Rank test used because their results were non-parametric. Comparisons between different group hormones were were performed with Friedman test. 5. Result and discussion This study evaluated the relation between the pregnancy hormones and alpha particles deposited in the blood of women. Blood samples containing LH, FSH, HCG, Progesterone and estradiol were analysed and irradiated. (Radium source: activity 5 µci). The results showed a relationship between the blood control samples before irradiation and postirradiation, as the levels of FSH were decreased from 6.75 IU/L to 2.94 IU/L, and LH levels from 10.1 IU/L to 6.3 IU/L, HCG from 0.70 miu/ml to 0.23 miu/ml, Progesterone from 0.66 ng/ml to 0.27 ng/ml, and Estradiol from 46.7 pg/ml to 25.8 pg/ml. This relationship supports the hypothesis that the magnitude of changes in these markers is in fact indicative of radiation reaching the tissues. LH leads to ovarian production. Disruption of LH production in the ovaries results in both sterilization and loss of hormone production, because ovarian hormonal production is closely related to the presence of ova the follicle stimulating hormone (FSH) level is the basal maturation of the primary follicle. A high ovarian reserve usually indicates a good number of viable eggs present in ovaries while low ovarian reserve may indicate fewer available eggs. FSH level correlates inversely with the number of eggs present in ovaries. In this study, FSH was decreased by irradiation, thereby proving the effects of alpha particles on women fertility (novelty in this study). Moreover, the most common causes of impaired fertility among females are low ovulation and follicle stimulating hormone (FSH) secretion which in turn causes low estrogen secretion and decreased ovarian reserve simultaneously. An excess or deficiency of any hormone can cause the ovulation that leads to the infertility. Deficiencies of luteinizing hormone (LH) and follicle-stimulating hormone can lead to loss of menstruation accompanied by infertility in women (3), as shown in all tables of this study. HCG is the first hormonal factor with high concentrations during pregnancy as shown in table 3. However, HCG is decreases after irradiation, an inferred consequence of the impact of radiation on women fertility. Furthermore, ph in the blood changed after irradiation from 7.56 to 7.80, while the evaluated conductivity gave a mean value of 3.47 µs/cm. 1. Hormones and ph characteristics There were eleven subjects involved in this study. Hormones, ph of blood and urine were measured before and after radiation, as shown in table1. 23
4 2. Differences in hormone levels before and after radiation Significant differences were found before and after radiation. For LH, HCG, and Estradiol, the levels of these hormones significantly reduced after radiation, while elevation was observed for progesterone, FSH, blood and urine ph after radiation, as shown in table 2. Table 1: Hormone and ph characteristics of eleven subjects before and after irradiation Hormone Number of subjects Mean ± SD Median LH Before ± After ± FSH Before ± After ± HCG Before ± After ± Progesterone Before ± After ± Estradiol Before ± After ± ph Blood Before ± After ± Urine Before ± After ± Table 2: Difference in hormones and ph before and after irradiation Laboratory measurements Mean difference ± SD p value LH Before After ± < 0.001* Before After ± < 0.001* Progesterone Before After ± < 0.001* Laboratory measurements Ranks Number Z value p value FSH Before After Negative ranks ** Positive ranks 11 Ties Total Estradiol Before After Negative ranks ** Positive ranks 0 Ties Total
5 Table 3 Differences in changed hormones Changes in hormones Mean Rank Chi-square df p value LH < FSH 3.41 HCG 1.86 progesterone 1.14 estradiol 5.00 Conclusion This research evaluated the relation between women fertility and hormone markers of ovarian reserve. The study observed disparity in pregnancy hormone levels after exposure to radiation (alpha particles). All the pregnancy hormones showed significant differences (p< 0.001) before and after irradiation. Thus proving the negative effects of alpha particles on women infertility. Exposure to radiation, like alpha particles results in hormonal imbalance and changes in the hormone parameters, clearly indicating that all the hormones tested were affected by radiation and impact the fertility in women, resulting in abnormal ovulation, impaired fertility and infertility in women in Iraqi Kurdistan region. Therefore, a new concept and technique were presented in this research to study the effects of alpha particles on the pregnancy hormones. References 1- Robert S. M., M., Thomas L. T. (2002). The Evaluation of Infertility. American Journal Clinical Pathology. Review.117, 95-S Sonya M. Schuh-H., Nicholas A. J., Mitchell P. R.,Barbara S., Marcelle I. C., & Renee A. R. P. (2011) Genetic variants and environmental factors associated with hormonal markers of ovarian reserve in Caucasian and African American women. Human Reproduction, Vol.0, No.0 pp Anna N. Y. et al. (2012). Prevalence and Sociodemographic determinants of Infertility, Success of Infertility Treatments and Health of Treated Women. School of Medicine of the University of Tampere. Medisiinarinkatu 4- Fady I. S. David B. S., Jodi M.D. & Flaws. A. (1998). Environmental toxicants and female reproduction fertility and sterility 70, NO. 4, American Society for Reproductive Medicine Published by Elsevier Science Inc. U.S.A. 5- Martin G. B., et al., (1983). Effects of oestradiol, progesterone and rostenedione on the pustule secretion of luteinizing hormone in ovariectomized ewes during spring and autumn J Endocrinal February 96: U. S. (2008). Beyond Infertility Polycystic Ovary Syndrome (PCOS) National Institutes of Health Eunice Kennedy Shriver National Institute of Child Health and Human Development NIH Pub. No UNSCEAR. (1993). Sources and effects of ionizing radiation report to the general assembly with scientific annexes, 8- Emma Derbyshire. (2011). Nutrition and Fertility. Nutrition in the Childbearing Years, First. Edition. Emma Derbyshire. Published by Blackwell Publishing, 9- Jessie Z.-J. C., Penelope M. S., Shaun P. B., & Rosemary J. K., (2012). Vessel remodeling, pregnancy hormones and extra villous trophoblast function. Molecular and Cellular Endocrinology 349, pp Richard L. et al. (1999). Fertility testing. Tadiobiology and physics of brachytherapy. Radiation oncology, the cleveland clinic foundation, cleveland, 11- Robert J. N. (2002). Fertility testing. Australian Prescriber. Vol. 25 No.2, Hongxia L. Steven T. N., Jiangang C., Heather E. T., James W. O., & Bill L. L. (2001). Differences in hormonal characteristics of conceptive versus non conceptive menstrual cycles fertility and sterility. American Society for Reproductive Medicine Published by Elsevier Vol. 75, No Mamdoh A. E. et al., (2011). Early maternal serum β-human chorionic gonadotropin measurements after ICSI in the prediction of longterm pregnancy outcomes: A retrospective cohort analysis Original Article J Clin Med Res;3(1):30-35Press. 25
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