doi 10.15296/ijwhr.2017.20 http://www.ijwhr.net doi 10.15296/ijwhr.2015.27 OpenOpen Access Original Review Article

InternationalInternational Journal Journal of Women’s of Women’s Health Health and Reproduction and Reproduction Sciences Sciences Vol.Vol. 3, No.5, No. 3, July 2, April 2015, 2017, 126–131 107–111 ISSNISSN 2330- 4456 2330- 4456

InvestigatingWomen on the theOther Success Side of Rate War of and Sex Poverty: Selection Its Effectin Cycles ofon Intrauterine the Health of InseminationReproduction of Sperm Using an Albumin GradientAyse Cevirme1, Yasemin Method Hamlaci2* , inKevser Infertile Ozdemir2 Couples Referring to the OmidAbstract Persian Gulf Center of Bushehr, Iran War and poverty are ‘extraordinary conditions created by human intervention’ and ‘preventable public health problems.’ War and Shahnazpoverty have Ahmadi many negative1, Elham effects Rahmani on human2*, Niloofarhealth, especially Motamed women’s3,4, health.Fatemeh Health Sadeghi problems5 arising due to war and poverty are being observed as sexual abuse and rape, all kinds of violence and subsequent gynecologic and obstetrics problems with physiological and psychological courses, and pregnancies as the result of undesired but forced or obliged marriages and even rapes. Certainly, Abstractunjust treatment such as being unable to gain footing on the land it is lived (asylum seeker, refugee, etc.) and being deprived of Objectives:social security, Sex citizenship selection rightsis an importantand human concernrights brings for someabout couples.the deprivation One of of the access relatively to health more services simple and and of inexpensiveprovision of methods, whichservice does intended not needfor gynecology to use toxic and chemicals obstetrics. to The separate purpose the of Y this and article X sperms, is to address is the effectsEricsson of warmethod. and poverty Therefore, on the the health aim of this study isreproduction to evaluate ofthe women results and of sexto offer selection scientific in infertilecontribution couples and bysolutions. using the albumin gradient method. MaterialsKeywords: and Poverty, Methods: Reproductive The present health, study War is a quasi-experimental clinical trial study, conducted on 80 infertile couples. After the induction of and a vaginal sonography, sperm samples were prepared using the albumin gradient method, after which intrauterine insemination (IUI) was conducted. The rate of successful sex selection was then calculated. IntroductionResults: Among 80 cases of performed IUI, 22 cases (27.5%) werethought successful, that amongsevere whichmilitary 3 cases conflicts (8.3%) in suffered Africa miscarriagesshorten and Throughout19 cases (23.7%) the historywere associated of the world, with athe successful ones who pregnancy. had Thethe success expected rate lifetime of sex selection for more was than 59.1% 2 years. in successful In general, pregnancies. confrontedIn general, the sexbitterest selection face success of poverty rate wasand higher war had for althe- maleWHO gender; had as acalculated result, the that success 269 ratesthousand in 68 peoplerequests had for diedmale sex and ways12 requests been the for women. sexAs totaledknown 12poverty cases (17.6and war %) andaffects 1 case (3.8%),in 1999 respectively. due to the effect of wars and that loss of 8.44 mil- humanConclusion: health Albumin either directly gradient or isindirectly, an appropriate the effects sex determination of lion healthy method years in IUI of lifecycles had due occurred to a 23.7% (2,3). chance of pregnancy in thisinfertile condition women, on whilehealth the and success status rate of women is 59.1% in in the sex so determination.- Wars negatively affect the provision of health services. cietyKeywords: should Infertility, not be ignored. Insemination, This study Sex preselection, intends to Spermatozoacast Health institutions such as hospitals, laboratories and light on the effects of war and poverty on the reproductive health centers are direct targets of war. Moreover, the wars health of women. For this purpose, the face of war affect- cause the migration of qualified health employees, and ing the women, the problem of immigration, inequalities thus the health services hitches. Assessments made indi- Introductionin distribution of income based on gender and the effects cate thatdefects the areeffect present of destruction in one gender in the infrastructureof their children, of so they Failureof all these of a on woman the reproductive to become health pregnant of women during will one be year healthhave continues no option for 5-10 but toyears seek even gender after determination the finalization methods. ofaddressed. intercourse without the use of contraceptive methods of conflictsMoreover, (3). Duesometimes to resource the interestrequirements of some in the fertile re- couples is called infertility (1). Researchers believe that infertility structuringin a specific investments gender after has war,led them the share to determine allocated theirto child’s hasWar become and Women’s one Healthof the major social problems in the healthgender. has decreased In the (1).older days, when a male workforce was society,Famine, today. synonymous However, with infertility war and poverty,is one of is the clearer areas for in the more important than the female workforce, if a family medicalwomen; fieldwar means that can deep help disadvantages cure this problem such as fullusing de -data Mortalitieshad no and son, Morbidities the number of the family members could be fromstruction, relevant loss ofresearches future and (2,3). uncertainty for women. Wars The onesincreased who are beyond most affected eight fromin order wars toare achieve women andthe blessing areInfertility conflicts is that considered destroy families, as one societiesof the mostand cultures bitter and children.of a Whileson. That deaths is whydepending the statistics on direct of violencechildren af of- a family stressfulthat negatively life experiences affect the health for ofyoung community couples and because cause of fect thewere male high. population, Also, many the indirect families deaths suffered kill children, severely because theviolation many of psychiatric human rights. symptoms According that to the occur data of during World the womenof andthis eldersissue. more.There In is Iraq also between often 1990-1994,disagreement in- between Health Organization (WHO) and World Bank, in 2002 fant deaths had shown this reality in its more bare form diagnosis and treatment process. These are in addition spouses in newly established families over a child’s gender; wars had been among the first ten reasons which killed with an increase of 600% (4). The war taking five years tothe the most physical and caused problems, disabilities. which Civil disrupt losses aretheir at theway rate of life increasessome the prefer child sonsdeaths and under some age prefer of 5 by daughters. 13%. Also 47% The proven/ andof 90% the within consequent all losses problems (1). they experience in their of all establishedthe refugees methodsin the world of anddetermining 50% of asylum a child’s seekers gender is a relationshipsWar has many with negative their effects spouses, on humanfamily members,health. One friends of and displacedvery complicated people are phenomenon women and girlsbeyond and our44% imaginations. ref- andthese colleagues is its effect (4-12). of shortening Sometimes, the averagepsychological human impactslife. ugees andToday, asylum by using seekers medical, are children biology under and nutritional the age of sciences, ofAccording infertility to themay data also of WHO,affect thethe average cooperation human lifebetween is 18 (5).it has been has shown that parents can select their child’s patients,68.1 years the for medical males and team 72.7 and years medical for . staff. It is being As thegender. result Itof should wars and be armednoted thatconflicts, all of women these methodsare are On the other hand, an increased infertility rate relative and that only a laboratory method is definite (14- sometimesReceived 12 December leads 2014,couples Accepted to 25select April 2015,a specific Available genderonline 1 July (13) 2015. 16). Of course, this method is used more in couples with Sometimes, couples are not always infertile, but certain disabilities in one type of gender of children. 1Department of Nursing, Sakarya University, Sakarya, Turkey. 2Department of Midwifery, Sakarya University, Sakarya, Turkey. *Corresponding author: Yasemin Hamlaci, Department of Midwifery, Sakarya University, Sakarya, Turkey. Tel: +905556080628, Email: [email protected] Received 14 July 2016, Accepted 5 November 2016, Available online 16 December 2016

1Associate Professor, Department of Gynecologic Obstetric. Iran University of Medical Sciences, Tehran. Iran. 2Associate Professor, Department of Gynecologic Obstetric, Bushehr University of Medical Sciences, Bushehr, Iran. 3Department of Community Medicine, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran. 4The Persian Gulf Nuclear Medicine Research Center, The Persian Gulf Biomedical Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran. 5Student Research Committee, Bushehr University of Medical Sciences, Bushehr, Iran. *Corresponding Author: Elham Rahmani, Tel: 07733455409, Email:[email protected] Ahmadi et al

Materials and Methods 2. 0.5 ml of sediment was gently removed from each This study was a semi-experimental clinical trial, which tube. Due to the presence of albumin 10% and 37°C was conducted on infertile couples who wanted to for 30 minutes, motile sperms were deposited and a determine the gender of their child and who were referred sample was taken from the deposited part. to the Bushehr Omid Persian Gulf Infertility Center and 3. Four small Falcon tubes were used and 0.5 mL 20% Abolfazle from October 2015 to January 2016. All albumin and then 1 m of 12.5 albumin was added patients signed an informed written consent. The study was to each tube. Finally, 0.5 of sperm solution from approved by the ethical committee of Bushehr University the previous step was added and maintained for 45 of Medical Science. The inclusion criteria included a minutes at 37°C. patient being aged between 20 and 40 years, the opening Thirty-six hours after the HCG injection and preparation of the fallopian tubes on the HSG or laparoscopy, normal of seminal fluid on the day of intrauterine insemination semen analysis test, a request to determine the gender (IUI), the child’s sex was selected for each of the couples. and the desire to enter the study. The exclusion criteria Finally, the success rate of the sex selection by using the included medical illnesses such as thyroid disorders, albumin gradient technique for sperm separation in IUI increased prolactin and other endocrine diseases and cycles was studied. All the information, including the uterine problems, such as myoma and polyps. Primary couples’ ages, sperm motility, sperm morphology, number or secondary infertility and years of infertility were not of sperm, the expected and actual gender of the fetus and considered effective in this study; although those patients the number of cycles, was collected in the data form and who were eligible for in vitro fertilization (IVF) or intra- the analysis was carried out. cytoplasmic sperm injection (ICSI) were not included in this study. Results After the initial sonography and from the third to the Eighteen couples were included in the study. The minimum seventh day of menstruation, the patient was instructed and maximum age for the women ranged between 20 and to take 2 tablets of letrozole (Femara, Novartis, ) 40 years and their average age was 30.95 ± 5.02. There was orally, every day. On the eighth and ninth days, 75-150 at least one previous pregnancy in 63.3% of cases and a units of Gonal F (Gonal f, Serono, Aubnne, Switzerland) primary pregnancy only accounted for 36.7% of cases were subcutaneously injected. On the tenth day, a vaginal (Table 1). ultrasonography was done and then ultrasonographies A total of 85% (68 couples) desired to have a son, while were repeated every other day, depending on the growth 15% (12 couples) desired to have a daughter, respectively. of the follicles. When 1-3 follicles of 19-24 mm in size Among the 80 couples, 58 cases (72.5%) experienced an were seen during the transvaginal sonography, 10 000 unsuccessful IUI and 22 cases (27.5%) had a successful units of HCG (Pregnyl, Organon, Oss, Netherlands) were IUI, among which 3 cases (8.3%) suffered and 19 administered through an intramuscular injection (IM). cases (23.7%) experienced successful pregnancy (Table 2). Semen samples were taken 36 hours after this and the Considering the 22 successful IUI cases (including 3 following additional steps were carried out on the samples. cases of abortion), the success rate of sex selection was Semen samples were collected by using the self-excited 59.1%. Among the 19 cases of successful pregnancy, method in disposable sterile containers and then kept sex selection was successful in 13 cases (68.4%, 95% CI: at a temperature of 37°C.The status of the seminal fluid 43.44-87.42) and was unsuccessful in 6 cases (31.6%). macroscopic parameters, including volume, agglutination and consistency became clear by accessing the World Table 1. Demographic Of Infertile Couples Referring to Omid Persian Health Organization (WHO). Afterwards, concentration, Gulf Infertility Center for Sex Selection type and percentage of motility of sperms were determined Variable Mean ± SD Min Max using a Makler Chamber. Giemsa staining was used to investigate the sperm morphology and concentration of Age 30.95 ± 5.02 19 40 round cells and 100 sperms, along with non-sperm cells Number of previous pregnancies 1.05 ± 0.95 0 4 that were fragmented among them. Number of IUI cycles 1.18 ± 0.41 1 3 Two to three milliliters of seminal fluid was poured into a Fanconi tube and albumin-free (pre-heated) Hams Table 2. The Results of IUI for Sex Selection in Infertile Couples F10 medium was added in the same value. The mixture Referring to Omid Persian Gulf Infertility Center for Sex Selection was later centrifuged at 2000 rpm for 10 minutes. The Variable Number Percent supernatant was discarded and 2 mL of Hams was added into it. Sperms containing X and Y chromosomes were Expected boy 68 85 separated using albumin gradient in the following manner: Expected girl 12 15 1. Four milliliters of heated human albumin 10% (at the Obtained boy 15 18.75 laboratory temperature) was equally added into four Obtained girl 4 5 small Falcon tubes, into each of which 0.5 mL sperm IUI success 22 27.5 solution prepared beforehand was added and placed for 30 minutes at 37°C. Miscarriage 3 3.8

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However, the overall success rate of 13 out of 80 couples, an abortion; the sample size has been decreased and, in considering both the success rate in conducting IUI and turn, this affects the results obtained. In another study, pregnancy success rate, was 16.25% (95% CI: 8.24-26.18). Nasresfahani et al (18) studied 45 couples and, in another In all 80 cases, the success rate was higher for the male study by Khalili et al (19), the sample size was equal to 32 gender. As such, in 68 requests for a male gender and subjects. Thus, according to the results of various studies, 12 requests for a female gender, 12 cases (17.6 %) and 1 a smaller sample size is effective in achieving the desired case (3.8%) were successfully determined. Also, among results. 19 successful pregnancies, out of the 15 male-requested In the present study, the letrozole tablet, which is effective cases, 12 (80%) were successful, whereas out of the 4 in sex determination, has been used for patients on days 3 female-requested cases, 1 (25%) was successful. There was to 7 of their menstruation period. In his study, Silverman no significant difference between the two successful and et al (20) used clomiphene citrate during ovulation, which unsuccessful IUI groups in terms of the mean number of increases the chance of delivering female infants. Lighter previous pregnancies (P = 0.68), the percentage of sperm sperms (Y sperm) in the albumin gradient technique with normal morphology (P = 0.50) and the number of produces much faster cross-human albumin layers than cycles of IUI (P = 0.98; Table 3). However, the sperm count X sperms (HAS) and are deposited in test tubes (21). in successful IUI group was significantly higher than the Researchers believe that the albumin gradient technique unsuccessful group (P = 0.035). is a good method because it does not use chemicals to Considering the level of normal sperm by as much as 40 separate sperms and the fetus is not manipulated. On million, the percentage of sperms counted over 40 million the other hand, many researchers including (17,21,22) was not significantly different between the two groups confirmed the effectiveness of this method, especially (P = 1). Also, the two groups in terms of sperm motility in determining the male gender. Meanwhile, other (P = 0.34) and the number sperms, which moved forward researchers believed that this method was ineffective (P = 1), was not significantly different (Table 4). (17,23). However, Fugger’s study suggests that the health of the baby after delivery has been confirmed by this Discussion method (24). In subjects aged between 20 to 40 years, the In the present study, which was conducted to investigate results showed that there was no significant difference the success rate of the albumin gradient method in between successful and unsuccessful groups, which selecting the sex of the fetus, 80 infertile couples who was not referred to in other studies. Other findings that wanted to determine the sex of their unborn child were showed no significant differences between the successful entered into the study. Finally, 58 failed cases and 3 cases and unsuccessful groups included the percentage of sperm of abortion from the remaining 22 cases occurred. The with normal morphology. The number of cycles and sample size in this study was higher than that of other sperm count was the only factor that was different in the studies, which, in turn, creates a broader and more accurate two groups in this study and was not mentioned in other scope of information. Also, the possibility of errors in this studies. sample size is lower, as it is easier to achieve the desired The above-mentioned factors suggest the accuracy and results. However, in the study conducted by the Center of extent of our study, which are indicative of the superiority (17) using the Ericsson method, 18 patients of our study compared with other studies. Another were studied, regardless of the 4 cases that resulted in factor, which was investigated in the present study, was

Table 3. The Comparison of 2 Groups of Patients in Infertile Couples Referring to Omid Persian Gulf Infertility Center for Sex Selection IUI Successful IUI Unsuccessful Variable t/z P Value (Mean ± SD) (Mean ± SD) Age 31.31±3.35 30.88±5.29 -0.27 0.78 Number of previous pregnancy 0.92± 0.86 1.07±0.97 -0.41 0.68 Number of IUI cycles 1.23±0.59 1.16±0.37 -0.02 0.98

Table 4. The Comparison Of the 2 Groups of Patients Regarding Quality Indicators of Sperm Analysis Infertile Couples Referring to Omid Persian Gulf Infertility Center for Sex Selection

Variable Successful Group Unsuccessful Group χ2 P Valuea Normal 12 (16.2%) 62 (83. 8%) Sperm count - 1 Abnormal 1 (16.7%) 5 (83.3%) Normal 13 (18.1%) 59 (81.9%) Sperm motility - 0.34 Abnormal 0 8 Normal 1 (16.5%) 66(83.5%) Forward movement - 1 Abnormal 0 1 a Fisher exact test.

International Journal of Women’s Health and Reproduction Sciences, Vol. 5, No. 2, April 2017 109 Ahmadi et al the history and number of pregnancies, which was not support and Bushehr Omid Persian Gulf infertility clinic referred to in other studies, except in the study of Khalili staff (Mrs. Yazdanikhah, Miss Kiamarci, Mrs. Bakhtiari et al (19), and this factor is also important in achieving the for data collection). This data was collected from Miss desired result. The reproductive ability was another factor Sadeghi’s thesis. that was referred to only in the study by Nasresfahani et al (18), who showed that 80% and 54.4% of infertile and References fertile women could achieve their desired results. Among 1. Ott MA, Sucato GS, Braverman PK, et al. Contraception the factors that were studied include the rate of successful for adolescents. 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