Impact of Radio Frequency Radiation (RFR) emitted from Cell Phone on Human Semen Quality

Impact of Radio Frequency Radiation (RFR) emitted from Cell Phone on Human Semen Quality | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Impact of Radio Frequency Radiation (RFR) emitted from Cell Phone on Human Semen Quality Ajeet -, Rakhi Singh, Shrawan K Trivedi, Rajeev Singh, Abhimanyu Kumar Jha, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4162568/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract A prospective study has been conducted to find out the impact of cell phone on the semen quality and hormonal profile of the men. After fulfilling the exclusion and inclusion criteria 421 healthy male volunteer’s aged 18–45 years were divided in to three groups depending on duration of cell phone use. Group-1 highly exposed group, Group-2 moderately exposed group, Group-3 control group. Subjects were asked to visit clinic every year for semen and blood sample collection up to three years. Semen physical characteristics and blood testosterone, T4 and TSH levels were estimated. Semen volume, rapid forward linear progression motility, moderate linear progression motility, sperm concentration, sperm density, percentage of normal and live sperm decreased in Group-1 in comparison to Groups-2 & 3 and trend continued till 3rd year. Sperm head abnormalities were higher in Group-1 than the Groups − 2 & 3 and trend continued till 3rd year. Slight decrease in levels of testosterone, significant increase in T4 and decrease in TSH were observed in Group-1 and similar trend was noticed till 3rd year. Present data indicate that the increased duration of cell phone use is adversely influencing the semen quality and hormone levels in men which may lead to infertility. EMF Semen Quality RFR Infertility Reproduction Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction The male infertility has been considered serious problem around the globe. The scientists have been working from many decades to determine the possible causes of male infertility. According to studies conducted both in-vitro and in-vivo, numerous factors are responsible for infertility in humans [14,18,42,35,3]. One of the important factors that has recently been reported to have an adverse impact is Radio Frequency Radiation (RFR) emitted from cell phones.RFR is subset of electromagnetic field and its frequency ranges from 3 KHz to 300 GHz. The electromagnetic radiations have been widely used in defense as well as in civilian sectors around the world. Radar surveillance defense communication links and terminals, high voltage power lines, large telecommunication facilities, commercial satellite communication, television broadcasting, geosynchronous satellite ground stations and microwave terrestrial link system are only a few examples of major development projects. Under civilian sector, cellular phones are one of the common sources of electromagnetic radiation. In India, the usages of cellular phones were initiated in 1994, and in 2001there were over 3.58 million subscribers using cell phones nationwide. Afterwards the number of subscribers dramatically expanded, reaching 584.32 million in 2010 and 1142.02 million by March 2022 in the country [49]. This technology was introduced without a thorough knowledge of the nature of these radiations or prior detailed discussion among the scientific community. The fixed broadcast facilities often located in residential areas, schools etc., additionally increased the potential exposure without paying any attention to the possible harm to the residents of the area. Due to increasing usage of cell phones in the country and decreasing cost of mobile phone, the levels of exposures to RFR have increased. Life was unimaginable without a cell phone during the duringthe COVID-19 pandemic. However, the majority of the people were unaware of the adverse impact of RFR emitted from cellular phones. It has been reported that RFR emitted from cell phone decrease the sperm counts, sperm motility, sperm viability and sperm morphology [3]. Agarwal et al 2008 reporteddecrease quality of semen in cell phone users due to the impact of RFR on the sperm count, viability, motility and normal morphology [5]. This group has also noticed that RFR has negative impact on oxidative stress in semen of male subjects [34]. It was noticed that human semen samples exposed to EMR showed a significant decrease in sperm motility and viability [51]. Gutschi T et al 2011 reported negative impact on sperm quality [20]. The chronic exposure to RFR emitted from cell phone causes a significant decrease in protein kinase C and total sperm count along with increase apoptosis in male wistar rats. The significant reduction has been noticed in testicular size, weight and in sperm counts in male wistar rats [25,26]. EMF radiation has been reported to significantly decrease sperm count and sperm viability along with the decrease in seminiferous tubule diameter and degeneration of seminiferous tubules in mice [43]. The studies conducted so far were unable to establish conclusive information about the safety or risk of RFR emitted by cell phones. These investigations did however highlight RFR’s possible biological impacts. The majorities of the relevant studies were conducted in isolation and were not community based. Therefore, in this community-based study, efforts have been made to better understand the adverse effects of radio frequency radiation emitted from cell phones on semen quality of North Indian population. Materials and Methods Sample Size The sample size for the study has been calculated as 385 with the confidence level of 95% that the real value is within ± 5% of the population proportion (50%). Keeping dropout rate in the mind, the 600 male subjects have been enrolled after fulfilling exclusion and inclusion criteria’s under various study groups. Total 600 healthy male volunteers age from 18–45 years age fulfilling the exclusion and inclusion criteria residing minimum 300 meter away from the cell phone tower in Delhi-NCR have been enrolled and followed during 2019 to 2023 under the study. These subjects were divided in following three groups depending on duration of their cell phone use. Group-1: Heavy Exposure Male Group (HEMG) which included the subjects who were exposed to EMF/RFR for more than four hours per day. Group-2: Moderate Exposure Male Group (MEMG) which included the subjects who were exposed to EMF/RFR less than 4 hour/day but not less than two hours/day. Group-3: Control Male Group (CMG) which included the subjects who were not exposed to any EMF/RFR or less than two hours/day. The demographic profiles of all the subjects of all three groups were recorded through a prescribed proforma. The subjects were then asked to report at the Abalone Clinic Maternity & Fertility Center, Noida, Uttar Pradesh, India for semen and blood collection. An informed written consent was obtained from all the participants. All subjects were advised to maintain three days abstinence before coming to the clinic for semen collection. Semen samples were collected by masturbation from all the subjects in a sterile plastic container in a room adjacent to the laboratory and kept at 37 0 C for 30 min for liquefaction. Liquefaction time of semen and viscosity were measured immediately after the collection of semen. The volume and pH of semen, sperm motility, sperm count, morphology, vitality and agglutination were assessed in liquefied semen samples, according to the methods described in the WHO laboratory Manual (WHO, 2010–5th edition). All experimental protocols were approved by the Institutional Ethics Committee on Human Research vide their letter No. IEC-HR/2019/01 dated 19/08/2019 and all research was performed in accordance with relevant guidelines/regulation. The levels of testosterone, T4 and TSH in the blood samples of all the subjects were estimated using ABBOTT Ireland kits for Chemiluminescent Microparticle Immunoassay (CMIA). All the enrolled subjects were asked to visit at the clinic after every year for semen collection till 3rd year. Out of total 600 enrolled subjects, 179 subjects were lost to follow-up and hence 421 enrolled subjects have been followed and also their semen samples were collected from 1st to 3rd year. The status of annual follow-up of these subjects’ group-wise, is given below under Table–1. Table-1: Group-wise Status of Annual Follow-up Groups Visit-1 Visit-2 Visit-3 G-1 (HEMG) 130 130 130 G-2 ( MEMG) 188 188 188 G-3 (CMG) 103 103 103 Total 421 421 421 Statistics and Data Science Window-based software was designed and developed using dot net platform and data collected was entered in this window-based software. The entered data was imported in the excel sheet as well as SQL 2008 R2 software. To avoid duplicate entries, missing values, errors, etc. the data mining was carried out using the different types of methods and quarries of SQL 2008 R2 before analyzing the data using exploratory data analysis. Analysis has been performed by using Rstudio version 4.1.2 (01-11-2021) of the R Foundation for Statistical Computing. Tidyverse, dplyr packages were used for exploratory analysis of all parameters between the Highly Exposed, Moderate Exposed and Control subjects. Exploratory Data Analysis (EDA) has been used to help to identify the above said errors and also help to understand the trend within the variables. The exploratory factor analysis was done to find out the correlation of variables using different methods of R studio package which help to short the data to a small set of summary variables. It was also helped in identifying the structure and the relationship between the variables. After that, statistical model was used to extract the meaningful information from different variables for conclusion. MANOVA has been used for subsequent analysis. Microsoft Word and Excel have been used to generate tables, etc. Results Comparison of all semen physical parameters among all the three groups was done by using descriptive analysis. Summary of descriptive analysis including Mean ± SD is given under Supplementary Table-1. No difference was observed in appearance, pH of the semen, viscosity and agglutination of the semen among all the three groups. Similarly, no significant variation was noticed in these parameters from 1st visit to 3rd visit in all three respective groups. But the semen volume decreased in highly exposed group in comparison to moderate and control group which is statistically significant. Similarly, semen volume is also decreasing in the subjects from 1st to 3rd visit in all the study groups (see at Fig: − 1). In highly exposed group, both rapid forward linear progression motility and moderate linear progression motility were decreasing in comparison to moderate and control groups which is statistically significant. Similarly, both rapid forward and moderate linear progression motility were decreasing from 1st to 3rd visit in all the study groups which is also statistically significant (see at Fig: − 2). The percentage of non-progression and immotile sperm is increasing in highly exposed group in comparison to moderate and control groups. Similarly, the percentage of non-progression and immotile were increasing from 1st to 3rd visit in all the study groups which is also statistically significant (see at Fig: − 3). Both sperm concentration (million/ml) and sperm density (million/ejaculate) were decreasing in highly exposed group in comparison to moderate and control group and trend of decreasing was notice in subsequent follow visits i.e. 1st to 3rd in all the study groups. The decreasing trend in sperm concentration (million/ml) was statistically significant but decreasing trend in sperm density (million/ejaculate) was not statistically significant (see at Fig: − 4). The percentage of normal sperm and percentage of live sperm both are decreasing in highly exposed group in comparison to moderate and control group and decreasing trend was notice from 1st to 3rd visit in all the study groups (see at Fig: − 5). The percentage of normal sperm has shown statistically significant. On the other hand, percentage of abnormal sperm was increasing in highly exposed group in comparison to moderate and control group and similar trend was notice during the follow up visits in all the study groups which is also statistically significant (see at Fig: − 6). All type of abnormalities in the head region of sperm (Normal Head (%), Pyriform Head (%), Amorphos Head (%), Tapering Head (%), Pin Head (%), Round Head (%), Small Head (%), Double Head (%), Large Head (%) were noticed in highly exposed group which were higher than the moderate and control group and similar trend was noticed during the subsequent visits 1st to 3rd in all the study groups which is also statistically significant. Among the various types of head in this study the percentage of amorphous head were higher other than the other type of head in highly exposed group in comparison to moderate and control groups and similar trend was noticed from 1st to 3rd visit in all the study groups (see at Supplementary Fig: − 1 to 5). The percentage of sperm having midpiece, principal piece defect and coil tail defect was higher in highly exposed group in comparison to the moderate and control groups and similar trend was noticed during the subsequent visits from 1st to 3rd in all the study groups which is also statistically significant (see at Fig: − 7). The slight decrease in serum testosterone was observed in highly exposed group in comparison to the moderate and control groups and similar trend was noticed during the subsequent visits 1st to 3rd in all the study groups but levels of testosterone vary within the normal range which is statistically not significant (see at Supplementary Fig: − 6). The level of T4 in highly exposed group was showing increasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups which is statistically significant (see at Supplementary Fig: − 7). The level of TSH in highly exposed group was showing decreasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups (see at Supplementary Fig: − 8). The datasets used and/or analysed during the current study available from the corresponding author on reasonable request. Discussion In the present community-based study, we analyzed impact of RFR emitted from cell phone on adult human semen quality. Results clearly indicated that there was a decrease in semen volume, linear progressive motility, sperm concentration, sperm density and percentage of normal sperm in highly exposed group in comparison to moderate and control groups. This decrease trend in all these parameters was very transpicuous from 1st year to 3rd year and it was statistically significant. On the other hand, percentage of abnormal sperm, non-motile sperm, abnormal head, midpiece and tail defects significantly increased in highly exposed group in comparison to moderately exposed and control groups with an increasing trend from 1st to 3rd year. These observations are in compliance with earlier reports [14,18,3,4,5,15,24,2,51]. Similar observations have also been reported in animal studies [27,30]. But Malyapa et al 1997 and Dasdag et al 2003 did not find adverse effect of cell phone radiation on mouse fibroblasts and sperm count, morphology, histology of rat’s testes respectively [31,13]. But it is not correct to compare a rat model to humans. This is because of the facts that rat testes are smaller in size, nonpendulous scrotum and it can migrate between the abdomen and scrotum in the inguinal canal [10]. In the present study, slight decrease in serum testosterone was noticed in highly exposed group in comparison to control and moderate groups which is statistically not significant and this trend continued during the subsequent visits 1st to 3rd in all the study groups. On the other hand, exposure to microwave have been reported to disrupts the seminiferous tubules and reduces the Leydig cell population and testosterone levels in rats [24]. It has been reported that testosterone is required for maintenance of structural morphology and physiology of seminiferous tubules, spermatogenesis and formation of spermatozoa [44,48]. Hence any changes in the concentration of testosterone will result in adverse effect on semen quality [24]. In the present study the levels of T4 in highly exposed group were showing increasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups which is statistically significant. On the other side the level of TSH were decreasing in highly exposed group in comparison to moderately exposed and control group. Delay in spermatogenesis with maturation arrest, no pachytene spermatocytes, a decrease in seminiferous tubule diameters, an impairment of the mitochondrial activity, and a reduction in lipid concentration have been reported in hyperthyroid rats [38]. Choudhury et al ( 2003 ) reported an alteration of antioxidant systems as catalase is upregulated while GPx is downregulated in hypothyroid rats [12]. Whereas levothyroxine administration in ram have been reported to cause reduction in sperm motility and testis weight [11]. Rijntjes et al ( 2008 ) concluded that hyperthyroidism delays Leydig cell development and adversely affects spermatogenesis in rats [40]. In thyrotoxicosis patients, the high level of circulating thyroid hormones has been reported to results in asthenozoospermia in more than half of the patients, oligozoospermia and teratozoospermia in about 40% of thyrotoxic patients [1]. These abnormalities are frequently associate with a reduced semen volume (hypoposia) [1]. Hence, reduced sperm concentration &motility, and increase in abnormal morphology along with an overall decrease in semen volume have been reported to be the main semen alterations of thyrotoxic male patients [47].RW Hudson et al (1992) reported lower progressive forward motility in adult men with thyrotoxicosis due to Graves’ disease compared with euthyroid controls [41]. Krassaset al ( 2002 ) observed that hyperthyroid patients have a lower sperm motility compared to euthyroid controls [29]. As it is known that in normal man, 2% of testosterone is free, 44% is bound to sex hormone-binding globulin (SHBG), and the remaining is bound to albumin and other proteins. This is free- and albumin-bound portions testosterone which is called as bio available testosterone [8,19,53]. It has been reported that in hyperthyroidism, total testosterone and SHBG levels are increased [23,37] and free Testosterone is transiently reduced or may be within normal range [22] but mean basal bio-available Testosterone level is decreased [50]. In few thyrotoxic male subjects elevated circulating estrogen level have been reported which is because of enhanced production rate of the estrogen and increment of peripheral conversion of androgen to estrogen [39]. These reports clearly indicate that if it is confirmed hyperthyroidism then despite increased or normal total testosterone level, thyrotoxic men may have androgen deficiency due to reduced free and bioavailable testosterone and increase estrogen level. Alterations in these hormones can cause gynecomastia (24%) [1], decreased libido (70%) [52], erectile dysfunction (56%) [39], spider angiomas, spermatogenetic dysfunction [28] and reduced sperm count and/or motility. The elevated levels of thyroxin may be provoking oxidative stress on sperm including hyperthyroidism associated hyperestroginemia, enhanced nitric oxide synthase gene expression with nitric oxide overproduction following increase of cytokines stimulating ROS generation, increase turnover of mitochondrial protein, mitoptosis and increased rate of thyroid hormone induced lipolysis [36]. Several studies have been conducted to investigate the effect of RFR emitted from GSM mobile phone on levels of thyroid hormones. Baby et al ( 2017 ) conducted a cross sectional study in South India and reported a significant correlation between the total radiation and the TSH values among both individuals with or without a family history of thyroid dysfunction [7]. In a systematic review (Asl JF et al 2019 ) three studies reported decrease in TSH level in the subjects exposed to radiation whereas in one study an increase in TSH level was noticed [6]. Asl JF et al ( 2019 ) concluded that exposure to microwave radiations could cause cellular stress and increased thyroid gland temperature which can adversely affect the iodine uptake by the thyroid gland leading to thyroid gland disfunction [6]. In another cohort study that prolonged exposure to RFR resulted in a significant (P < 0.01) decrease in the T3 and T4 levels of the exposed subjects, indicating a possible thyroid dysfunction [16]. Mortavazi et al ( 2009 ) reported high TSH, low T4 and normal T3 concentrations in excessive mobile phone users. The authors concluded a possible hypothyroid state and thyroid dysfunction which seems to be directly proportional to the degree of duration and intensity of RFR exposure [32]. The study also concluded a possible harmful effect of cell phone RFR on the hypothalamic-pituitary-thyroid (HPT) axis [32]. To find out the effects on cell phone radiation on the thyroid gland function Bergamaschi et al ( 2004 ) conducted a study therein he divided the subjects into various groups based on their frequency of cell phone use [9]. It was reported that the subjects who had conversations on their mobile phones for > 33 hours per month had a 9.9% prevalence of low TSH values compared to a prevalence of 6% in the group who talked on their mobile phones for < 19 hours per month (P < 0.05). Based on an exploration study on this subject, Muacevic et al (2021) reported that the mobile phone radiation could negatively affect thyroid gland function and hormone production directly by disrupting the thyroid gland tissue or indirectly by disrupting the Hypothalamic-Pituitary-Thyroid (HPT) axis [33]. Current literature has established that mobile phones can exert their deleterious effects on the thyroid gland through thermal and non-thermal effects by stimulating cellular receptors and causing severance in the microtubules between cells [45]. The non-thermal effect of RFR leads to Reactive Oxygen Species (ROS) formation and accumulation of heavy metals within the cells [45,46]. It has also been reported that RFR may stimulate NADPH oxidase present on the plasma membrane of the cells which leads to ROS formation and apoptosis induction [17]. This oxidative stress may also disrupt the cellular Ca2 + ion pumps, transporters, and binding proteins [17]. In addition, RFR itself may also act on Ca2 + pumps that lead to Ca2 + efflux from cells and cellular apoptosis [17]. Other cellular stressors may be responsible for this disruption include increased cortisol levels and enzymes involved in thyroid homeostases, such as anti-thyroperoxidase, liver deiodinase, and ornithine carboxylase [21]. Conclusion Our data and available literature are clearly indicating that heavy use of cell phone adversely influencing the Hypothalamic-pituitary-thyroid axis and testes resulting in poor quality of semen in the form of decrease in semen volume, sperm concentration, motility, percentage of live and normal sperm and increase in percentage of abnormal sperm, abnormal sperm head, trunk & tail. Such adverse effects have been reported to be responsible for infertility due to the factors described under discussion. Declarations Acknowledgement: We thankfully acknowledge the support and assistance provided by the technical staff of Abalone Clinic Maternity & Fertility Centre, Noida, UP, India and financial support for undertaking all the investigations at the clinic. Funding: N/A Conflicts of interest/Competing interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Data availability: Data is available and will be provided on the request. Code availability: Code is available and will be provided on the request. Authors' contributions: Ajeet was collected the data and entered the data. Ravi Datta Sharma, Abhimanyu Kumar Jha, Rakhi Singh, Rajeev Singh and Radhey Shyam Sharma were designed the manuscript. Ajeet, Shrawan K Trivedi, Ravi Datta Sharma and Radhey Shyam Sharma were analyzed the data and wrote the manuscript. 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Plasma estrone, prolactin, neurophysin, and sex steroid-binding globulin in chronic alcoholic men. 24(9):1015-1019 (1975). Veerachari S B & Vasan SS. Mobile phone electromagnetive waves and its effect on human ejaculated human semen. Int. J. Infertility Fetal Med. 3:15-21 (2012). Velázquez E M & Bellabarba G A. Effects of thyroid status on pituitary gonadotropin and testicular reserve in men. Arch Androl. 38(1):85-92 (1997). Veldhuis J D. Male hypothalamic-pituitary-gonadal axis. In: Lipshultz LI, Howards SS, Ed. Infertility in the male, 3rd ed. Mooby-year Book St. Louis Mo. 23-58 (1997). Additional Declarations No competing interests reported. Supplementary Files Supplementary1.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4162568","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":291376890,"identity":"c5256809-15b5-4a27-8004-29777abfab4d","order_by":0,"name":"Ajeet -","email":"","orcid":"","institution":"Indian Council of Medical Research","correspondingAuthor":false,"prefix":"","firstName":"Ajeet","middleName":"","lastName":"-","suffix":""},{"id":291376891,"identity":"2b2a436b-5384-4b07-a8d2-603089d49f68","order_by":1,"name":"Rakhi Singh","email":"","orcid":"","institution":"Abalone Clinic Maternity \u0026 Fertility Center","correspondingAuthor":false,"prefix":"","firstName":"Rakhi","middleName":"","lastName":"Singh","suffix":""},{"id":291376892,"identity":"c2da808d-cbc1-4fc2-845e-6cfb204861dd","order_by":2,"name":"Shrawan K Trivedi","email":"","orcid":"","institution":"Rajiv Gandhi Institute of Petroleum Technology","correspondingAuthor":false,"prefix":"","firstName":"Shrawan","middleName":"K","lastName":"Trivedi","suffix":""},{"id":291376893,"identity":"f9f77261-5879-4bde-a422-ad84b4e1e00e","order_by":3,"name":"Rajeev Singh","email":"","orcid":"","institution":"Jamia Millia Islamia (Central University)","correspondingAuthor":false,"prefix":"","firstName":"Rajeev","middleName":"","lastName":"Singh","suffix":""},{"id":291376894,"identity":"cd79c5c2-5294-4087-af4e-f337cdac9f83","order_by":4,"name":"Abhimanyu Kumar Jha","email":"","orcid":"","institution":"Sharda University","correspondingAuthor":false,"prefix":"","firstName":"Abhimanyu","middleName":"Kumar","lastName":"Jha","suffix":""},{"id":291376895,"identity":"ed6f468e-9e8a-4023-b8eb-fbd46d2e503c","order_by":5,"name":"Ravi Datta Sharma","email":"","orcid":"","institution":"Amity University","correspondingAuthor":false,"prefix":"","firstName":"Ravi","middleName":"Datta","lastName":"Sharma","suffix":""},{"id":291376896,"identity":"d2e6bf3f-6713-4b66-bc47-81123785eaf4","order_by":6,"name":"Radhey Shyam Sharma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzUlEQVRIiWNgGAWjYHACMzDJDyISCkjRItkA0mJAihaDA2CSCPXm7Ye3PfjYVmdvfH514ocHBgzy/GIH8GuROZNWbjiz7XDithtvN0sAHWY4c3YCfi0SDDlm0rxtBxLMbpzdANKSYHCbkBb+N2bSf0EOm3F28w/itEgAbWFsY2bcwN+7jUhbJJ6VSfacO5w44wbvNosEAwki/MKfvE3iR1mdPX//2c03f1TYyPNLE9ACBoxsIM1glRJEKAeDP0DMf4BY1aNgFIyCUTDSAAD1D0LdtnAl1AAAAABJRU5ErkJggg==","orcid":"","institution":"Sharda University","correspondingAuthor":true,"prefix":"","firstName":"Radhey","middleName":"Shyam","lastName":"Sharma","suffix":""}],"badges":[],"createdAt":"2024-03-25 10:27:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4162568/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4162568/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":54977281,"identity":"df034168-e88e-4cdc-84fc-9f4b25a8cb1f","added_by":"auto","created_at":"2024-04-19 13:05:48","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":238465,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSemen Volume (ml)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/026ce5d991f75d00fe43c66e.jpg"},{"id":54977734,"identity":"014117ac-8f63-486a-ab12-cc8dad94d41a","added_by":"auto","created_at":"2024-04-19 13:13:48","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":290217,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRapid Forward and Moderate Linear Progression (%)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/831e1e7e05ea9e12b1e5a839.jpg"},{"id":54977277,"identity":"56ceff4d-5f54-4bee-9783-c38ae3af993a","added_by":"auto","created_at":"2024-04-19 13:05:48","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":286737,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePercentage of Non-progression and Immotile Sperm\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/ce4a764c9955468e0ea43bf0.jpg"},{"id":54977278,"identity":"11711474-b984-49d2-8a3a-4df1655a1afe","added_by":"auto","created_at":"2024-04-19 13:05:48","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":281019,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSperm Concentration (million/ml) and Sperm Density (million/ejaculate)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/91fbaebffec11eb38df016a1.jpg"},{"id":54977735,"identity":"c63fd78d-6c06-4f80-9edd-97d3349a39b9","added_by":"auto","created_at":"2024-04-19 13:13:48","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":339795,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePercentage of Normal Sperm and Sperm Vitality (% of Live Sperm)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/93bca95a4276d23c85062e05.jpg"},{"id":54977283,"identity":"c7108a83-3357-4509-821d-b3028f8428a7","added_by":"auto","created_at":"2024-04-19 13:05:48","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":192578,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePercentage of Abnormal Sperm\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/f35c9c9f761073aba473350b.jpg"},{"id":54977284,"identity":"be8c83ea-d25e-4fd5-92ab-bbc1512eed7c","added_by":"auto","created_at":"2024-04-19 13:05:49","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":313177,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePercentage of Sperm having Midpiece, Principal Piece Defect and Coil Tail Defect\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/dc52b15273f109fa1823c0c1.jpg"},{"id":72267271,"identity":"e7a97ca9-a2f6-440a-84c2-203066d2c93c","added_by":"auto","created_at":"2024-12-24 12:31:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2387331,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/938d09c5-8566-4cc0-b678-bece4451ec95.pdf"},{"id":54978326,"identity":"03a44b79-ccbc-4712-8131-feea34178222","added_by":"auto","created_at":"2024-04-19 13:21:48","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":85556,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementary1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4162568/v1/099f21fac69ca5e06be247b7.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Impact of Radio Frequency Radiation (RFR) emitted from Cell Phone on Human Semen Quality","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe male infertility has been considered serious problem around the globe. The scientists have been working from many decades to determine the possible causes of male infertility. According to studies conducted both in-vitro and in-vivo, numerous factors are responsible for infertility in humans [14,18,42,35,3]. One of the important factors that has recently been reported to have an adverse impact is Radio Frequency Radiation (RFR) emitted from cell phones.RFR is subset of electromagnetic field and its frequency ranges from 3 KHz to 300 GHz. The electromagnetic radiations have been widely used in defense as well as in civilian sectors around the world. Radar surveillance defense communication links and terminals, high voltage power lines, large telecommunication facilities, commercial satellite communication, television broadcasting, geosynchronous satellite ground stations and microwave terrestrial link system are only a few examples of major development projects.\u003c/p\u003e \u003cp\u003eUnder civilian sector, cellular phones are one of the common sources of electromagnetic radiation. In India, the usages of cellular phones were initiated in 1994, and in 2001there were over 3.58\u0026nbsp;million subscribers using cell phones nationwide. Afterwards the number of subscribers dramatically expanded, reaching 584.32\u0026nbsp;million in 2010 and 1142.02\u0026nbsp;million by March 2022 in the country [49]. This technology was introduced without a thorough knowledge of the nature of these radiations or prior detailed discussion among the scientific community. The fixed broadcast facilities often located in residential areas, schools etc., additionally increased the potential exposure without paying any attention to the possible harm to the residents of the area.\u003c/p\u003e \u003cp\u003eDue to increasing usage of cell phones in the country and decreasing cost of mobile phone, the levels of exposures to RFR have increased. Life was unimaginable without a cell phone during the duringthe COVID-19 pandemic. However, the majority of the people were unaware of the adverse impact of RFR emitted from cellular phones. It has been reported that RFR emitted from cell phone decrease the sperm counts, sperm motility, sperm viability and sperm morphology [3]. Agarwal et al \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2008\u003c/span\u003ereporteddecrease quality of semen in cell phone users due to the impact of RFR on the sperm count, viability, motility and normal morphology [5]. This group has also noticed that RFR has negative impact on oxidative stress in semen of male subjects [34]. It was noticed that human semen samples exposed to EMR showed a significant decrease in sperm motility and viability [51]. Gutschi T et al \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2011\u003c/span\u003e reported negative impact on sperm quality [20]. The chronic exposure to RFR emitted from cell phone causes a significant decrease in protein kinase C and total sperm count along with increase apoptosis in male wistar rats. The significant reduction has been noticed in testicular size, weight and in sperm counts in male wistar rats [25,26]. EMF radiation has been reported to significantly decrease sperm count and sperm viability along with the decrease in seminiferous tubule diameter and degeneration of seminiferous tubules in mice [43].\u003c/p\u003e \u003cp\u003eThe studies conducted so far were unable to establish conclusive information about the safety or risk of RFR emitted by cell phones. These investigations did however highlight RFR\u0026rsquo;s possible biological impacts. The majorities of the relevant studies were conducted in isolation and were not community based. Therefore, in this community-based study, efforts have been made to better understand the adverse effects of radio frequency radiation emitted from cell phones on semen quality of North Indian population.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003eSample Size\u003c/h2\u003e\n\u003cp\u003eThe sample size for the study has been calculated as 385 with the confidence level of 95% that the real value is within \u0026plusmn;\u0026thinsp;5% of the population proportion (50%). Keeping dropout rate in the mind, the 600 male subjects have been enrolled after fulfilling exclusion and inclusion criteria\u0026rsquo;s under various study groups. Total 600 healthy male volunteers age from 18\u0026ndash;45 years age fulfilling the exclusion and inclusion criteria residing minimum 300 meter away from the cell phone tower in Delhi-NCR have been enrolled and followed during 2019 to 2023 under the study. These subjects were divided in following three groups depending on duration of their cell phone use.\u003c/p\u003e\n\u003cp\u003eGroup-1: Heavy Exposure Male Group (HEMG) which included the subjects who were exposed to EMF/RFR for more than four hours per day.\u003c/p\u003e\n\u003cp\u003eGroup-2: Moderate Exposure Male Group (MEMG) which included the subjects who were exposed to EMF/RFR less than 4 hour/day but not less than two hours/day.\u003c/p\u003e\n\u003cp\u003eGroup-3: Control Male Group (CMG) which included the subjects who were not exposed to any EMF/RFR or less than two hours/day.\u003c/p\u003e\n\u003cp\u003eThe demographic profiles of all the subjects of all three groups were recorded through a prescribed proforma. The subjects were then asked to report at the Abalone Clinic Maternity \u0026amp; Fertility Center, Noida, Uttar Pradesh, India for semen and blood collection. An informed written consent was obtained from all the participants. All subjects were advised to maintain three days abstinence before coming to the clinic for semen collection. Semen samples were collected by masturbation from all the subjects in a sterile plastic container in a room adjacent to the laboratory and kept at 37\u003csup\u003e0\u003c/sup\u003e C for 30 min for liquefaction. Liquefaction time of semen and viscosity were measured immediately after the collection of semen. The volume and pH of semen, sperm motility, sperm count, morphology, vitality and agglutination were assessed in liquefied semen samples, according to the methods described in the WHO laboratory Manual (WHO, 2010\u0026ndash;5th edition). All experimental protocols were approved by the Institutional Ethics Committee on Human Research vide their letter No. IEC-HR/2019/01 dated 19/08/2019 and all research was performed in accordance with relevant guidelines/regulation. The levels of testosterone, T4 and TSH in the blood samples of all the subjects were estimated using ABBOTT Ireland kits for Chemiluminescent Microparticle Immunoassay (CMIA). All the enrolled subjects were asked to visit at the clinic after every year for semen collection till 3rd year.\u003c/p\u003e\n\u003cp\u003eOut of total 600 enrolled subjects, 179 subjects were lost to follow-up and hence 421 enrolled subjects have been followed and also their semen samples were collected from 1st to 3rd year. The status of annual follow-up of these subjects\u0026rsquo; group-wise, is given below under Table\u0026ndash;1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable-1: Group-wise Status of Annual Follow-up\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Taba\" border=\"1\"\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGroups\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eVisit-1\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eVisit-2\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eVisit-3\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eG-1 (HEMG)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e130\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e130\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e130\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eG-2\u003c/strong\u003e (\u003cstrong\u003eMEMG)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e188\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e188\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e188\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eG-3 (CMG)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e103\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e103\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e103\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e421\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e421\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e421\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003eStatistics and Data Science\u003c/h2\u003e\n\u003cp\u003eWindow-based software was designed and developed using dot net platform and data collected was entered in this window-based software. The entered data was imported in the excel sheet as well as SQL 2008 R2 software. To avoid duplicate entries, missing values, errors, etc. the data mining was carried out using the different types of methods and quarries of SQL 2008 R2 before analyzing the data using exploratory data analysis.\u003c/p\u003e\n\u003cp\u003eAnalysis has been performed by using Rstudio version 4.1.2 (01-11-2021) of the R Foundation for Statistical Computing. Tidyverse, dplyr packages were used for exploratory analysis of all parameters between the Highly Exposed, Moderate Exposed and Control subjects. Exploratory Data Analysis (EDA) has been used to help to identify the above said errors and also help to understand the trend within the variables. The exploratory factor analysis was done to find out the correlation of variables using different methods of R studio package which help to short the data to a small set of summary variables. It was also helped in identifying the structure and the relationship between the variables. After that, statistical model was used to extract the meaningful information from different variables for conclusion. MANOVA has been used for subsequent analysis. Microsoft Word and Excel have been used to generate tables, etc.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eComparison of all semen physical parameters among all the three groups was done by using descriptive analysis. Summary of descriptive analysis including Mean\u0026thinsp;\u003cspan class=\"Underline\"\u003e\u0026plusmn;\u003c/span\u003e\u0026thinsp;SD is given under Supplementary Table-1. No difference was observed in appearance, pH of the semen, viscosity and agglutination of the semen among all the three groups. Similarly, no significant variation was noticed in these parameters from 1st visit to 3rd visit in all three respective groups. But the semen volume decreased in highly exposed group in comparison to moderate and control group which is statistically significant. Similarly, semen volume is also decreasing in the subjects from 1st to 3rd visit in all the study groups (see at Fig: \u0026minus;\u0026thinsp;1).\u003c/p\u003e\n\u003cp\u003eIn highly exposed group, both rapid forward linear progression motility and moderate linear progression motility were decreasing in comparison to moderate and control groups which is statistically significant. Similarly, both rapid forward and moderate linear progression motility were decreasing from 1st to 3rd visit in all the study groups which is also statistically significant (see at Fig: \u0026minus;\u0026thinsp;2).\u003c/p\u003e\n\u003cp\u003eThe percentage of non-progression and immotile sperm is increasing in highly exposed group in comparison to moderate and control groups. Similarly, the percentage of non-progression and immotile were increasing from 1st to 3rd visit in all the study groups which is also statistically significant (see at Fig: \u0026minus;\u0026thinsp;3).\u003c/p\u003e\n\u003cp\u003eBoth sperm concentration (million/ml) and sperm density (million/ejaculate) were decreasing in highly exposed group in comparison to moderate and control group and trend of decreasing was notice in subsequent follow visits i.e. 1st to 3rd in all the study groups. The decreasing trend in sperm concentration (million/ml) was statistically significant but decreasing trend in sperm density (million/ejaculate) was not statistically significant (see at Fig: \u0026minus;\u0026thinsp;4).\u003c/p\u003e\n\u003cp\u003eThe percentage of normal sperm and percentage of live sperm both are decreasing in highly exposed group in comparison to moderate and control group and decreasing trend was notice from 1st to 3rd visit in all the study groups (see at Fig: \u0026minus;\u0026thinsp;5). The percentage of normal sperm has shown statistically significant. On the other hand, percentage of abnormal sperm was increasing in highly exposed group in comparison to moderate and control group and similar trend was notice during the follow up visits in all the study groups which is also statistically significant (see at Fig: \u0026minus;\u0026thinsp;6).\u003c/p\u003e\n\u003cp\u003eAll type of abnormalities in the head region of sperm (Normal Head (%), Pyriform Head (%), Amorphos Head (%), Tapering Head (%), Pin Head (%), Round Head (%), Small Head (%), Double Head (%), Large Head (%) were noticed in highly exposed group which were higher than the moderate and control group and similar trend was noticed during the subsequent visits 1st to 3rd in all the study groups which is also statistically significant. Among the various types of head in this study the percentage of amorphous head were higher other than the other type of head in highly exposed group in comparison to moderate and control groups and similar trend was noticed from 1st to 3rd visit in all the study groups (see at Supplementary Fig: \u0026minus;\u0026thinsp;1 to 5).\u003c/p\u003e\n\u003cp\u003eThe percentage of sperm having midpiece, principal piece defect and coil tail defect was higher in highly exposed group in comparison to the moderate and control groups and similar trend was noticed during the subsequent visits from 1st to 3rd in all the study groups which is also statistically significant (see at Fig: \u0026minus;\u0026thinsp;7).\u003c/p\u003e\n\u003cp\u003eThe slight decrease in serum testosterone was observed in highly exposed group in comparison to the moderate and control groups and similar trend was noticed during the subsequent visits 1st to 3rd in all the study groups but levels of testosterone vary within the normal range which is statistically not significant (see at Supplementary Fig: \u0026minus;\u0026thinsp;6). The level of T4 in highly exposed group was showing increasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups which is statistically significant (see at Supplementary Fig: \u0026minus;\u0026thinsp;7). The level of TSH in highly exposed group was showing decreasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups (see at Supplementary Fig: \u0026minus;\u0026thinsp;8).\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analysed during the current study available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn the present community-based study, we analyzed impact of RFR emitted from cell phone on adult human semen quality. Results clearly indicated that there was a decrease in semen volume, linear progressive motility, sperm concentration, sperm density and percentage of normal sperm in highly exposed group in comparison to moderate and control groups. This decrease trend in all these parameters was very transpicuous from 1st year to 3rd year and it was statistically significant. On the other hand, percentage of abnormal sperm, non-motile sperm, abnormal head, midpiece and tail defects significantly increased in highly exposed group in comparison to moderately exposed and control groups with an increasing trend from 1st to 3rd year. These observations are in compliance with earlier reports [14,18,3,4,5,15,24,2,51]. Similar observations have also been reported in animal studies [27,30]. But Malyapa et al \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1997\u003c/span\u003e and Dasdag et al \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2003\u003c/span\u003e did not find adverse effect of cell phone radiation on mouse fibroblasts and sperm count, morphology, histology of rat\u0026rsquo;s testes respectively [31,13]. But it is not correct to compare a rat model to humans. This is because of the facts that rat testes are smaller in size, nonpendulous scrotum and it can migrate between the abdomen and scrotum in the inguinal canal [10].\u003c/p\u003e \u003cp\u003eIn the present study, slight decrease in serum testosterone was noticed in highly exposed group in comparison to control and moderate groups which is statistically not significant and this trend continued during the subsequent visits 1st to 3rd in all the study groups. On the other hand, exposure to microwave have been reported to disrupts the seminiferous tubules and reduces the Leydig cell population and testosterone levels in rats [24]. It has been reported that testosterone is required for maintenance of structural morphology and physiology of seminiferous tubules, spermatogenesis and formation of spermatozoa [44,48]. Hence any changes in the concentration of testosterone will result in adverse effect on semen quality [24].\u003c/p\u003e \u003cp\u003eIn the present study the levels of T4 in highly exposed group were showing increasing trend during the follow up visit from 1st to 3rd in highly exposed group in comparison to the moderate and control groups which is statistically significant. On the other side the level of TSH were decreasing in highly exposed group in comparison to moderately exposed and control group. Delay in spermatogenesis with maturation arrest, no pachytene spermatocytes, a decrease in seminiferous tubule diameters, an impairment of the mitochondrial activity, and a reduction in lipid concentration have been reported in hyperthyroid rats [38]. Choudhury et al (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) reported an alteration of antioxidant systems as catalase is upregulated while GPx is downregulated in hypothyroid rats [12]. Whereas levothyroxine administration in ram have been reported to cause reduction in sperm motility and testis weight [11]. Rijntjes et al (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) concluded that hyperthyroidism delays Leydig cell development and adversely affects spermatogenesis in rats [40].\u003c/p\u003e \u003cp\u003eIn thyrotoxicosis patients, the high level of circulating thyroid hormones has been reported to results in asthenozoospermia in more than half of the patients, oligozoospermia and teratozoospermia in about 40% of thyrotoxic patients [1]. These abnormalities are frequently associate with a reduced semen volume (hypoposia) [1]. Hence, reduced sperm concentration \u0026amp;motility, and increase in abnormal morphology along with an overall decrease in semen volume have been reported to be the main semen alterations of thyrotoxic male patients [47].RW Hudson et al (1992) reported lower progressive forward motility in adult men with thyrotoxicosis due to Graves\u0026rsquo; disease compared with euthyroid controls [41]. Krassaset al (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e) observed that hyperthyroid patients have a lower sperm motility compared to euthyroid controls [29].\u003c/p\u003e \u003cp\u003eAs it is known that in normal man, 2% of testosterone is free, 44% is bound to sex hormone-binding globulin (SHBG), and the remaining is bound to albumin and other proteins. This is free- and albumin-bound portions testosterone which is called as bio available testosterone [8,19,53]. It has been reported that in hyperthyroidism, total testosterone and SHBG levels are increased [23,37] and free Testosterone is transiently reduced or may be within normal range [22] but mean basal bio-available Testosterone level is decreased [50]. In few thyrotoxic male subjects elevated circulating estrogen level have been reported which is because of enhanced production rate of the estrogen and increment of peripheral conversion of androgen to estrogen [39]. These reports clearly indicate that if it is confirmed hyperthyroidism then despite increased or normal total testosterone level, thyrotoxic men may have androgen deficiency due to reduced free and bioavailable testosterone and increase estrogen level. Alterations in these hormones can cause gynecomastia (24%) [1], decreased libido (70%) [52], erectile dysfunction (56%) [39], spider angiomas, spermatogenetic dysfunction [28] and reduced sperm count and/or motility. The elevated levels of thyroxin may be provoking oxidative stress on sperm including hyperthyroidism associated hyperestroginemia, enhanced nitric oxide synthase gene expression with nitric oxide overproduction following increase of cytokines stimulating ROS generation, increase turnover of mitochondrial protein, mitoptosis and increased rate of thyroid hormone induced lipolysis [36].\u003c/p\u003e \u003cp\u003eSeveral studies have been conducted to investigate the effect of RFR emitted from GSM mobile phone on levels of thyroid hormones. Baby et al (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) conducted a cross sectional study in South India and reported a significant correlation between the total radiation and the TSH values among both individuals with or without a family history of thyroid dysfunction [7]. In a systematic review (Asl JF et al \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) three studies reported decrease in TSH level in the subjects exposed to radiation whereas in one study an increase in TSH level was noticed [6]. Asl JF et al (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) concluded that exposure to microwave radiations could cause cellular stress and increased thyroid gland temperature which can adversely affect the iodine uptake by the thyroid gland leading to thyroid gland disfunction [6]. In another cohort study that prolonged exposure to RFR resulted in a significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) decrease in the T3 and T4 levels of the exposed subjects, indicating a possible thyroid dysfunction [16]. Mortavazi et al (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) reported high TSH, low T4 and normal T3 concentrations in excessive mobile phone users. The authors concluded a possible hypothyroid state and thyroid dysfunction which seems to be directly proportional to the degree of duration and intensity of RFR exposure [32]. The study also concluded a possible harmful effect of cell phone RFR on the hypothalamic-pituitary-thyroid (HPT) axis [32].\u003c/p\u003e \u003cp\u003eTo find out the effects on cell phone radiation on the thyroid gland function Bergamaschi et al (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) conducted a study therein he divided the subjects into various groups based on their frequency of cell phone use [9]. It was reported that the subjects who had conversations on their mobile phones for \u0026gt;\u0026thinsp;33 hours per month had a 9.9% prevalence of low TSH values compared to a prevalence of 6% in the group who talked on their mobile phones for \u0026lt;\u0026thinsp;19 hours per month (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Based on an exploration study on this subject, Muacevic et al (2021) reported that the mobile phone radiation could negatively affect thyroid gland function and hormone production directly by disrupting the thyroid gland tissue or indirectly by disrupting the Hypothalamic-Pituitary-Thyroid (HPT) axis [33].\u003c/p\u003e \u003cp\u003eCurrent literature has established that mobile phones can exert their deleterious effects on the thyroid gland through thermal and non-thermal effects by stimulating cellular receptors and causing severance in the microtubules between cells [45]. The non-thermal effect of RFR leads to Reactive Oxygen Species (ROS) formation and accumulation of heavy metals within the cells [45,46]. It has also been reported that RFR may stimulate NADPH oxidase present on the plasma membrane of the cells which leads to ROS formation and apoptosis induction [17]. This oxidative stress may also disrupt the cellular Ca2\u0026thinsp;+\u0026thinsp;ion pumps, transporters, and binding proteins [17]. In addition, RFR itself may also act on Ca2\u0026thinsp;+\u0026thinsp;pumps that lead to Ca2\u0026thinsp;+\u0026thinsp;efflux from cells and cellular apoptosis [17]. Other cellular stressors may be responsible for this disruption include increased cortisol levels and enzymes involved in thyroid homeostases, such as anti-thyroperoxidase, liver deiodinase, and ornithine carboxylase [21].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eOur data and available literature are clearly indicating that heavy use of cell phone adversely influencing the Hypothalamic-pituitary-thyroid axis and testes resulting in poor quality of semen in the form of decrease in semen volume, sperm concentration, motility, percentage of live and normal sperm and increase in percentage of abnormal sperm, abnormal sperm head, trunk \u0026amp; tail. Such adverse effects have been reported to be responsible for infertility due to the factors described under discussion.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement:\u0026nbsp;\u003c/strong\u003eWe thankfully acknowledge the support and assistance provided by the technical staff of Abalone Clinic Maternity \u0026amp; Fertility Centre, Noida, UP, India and financial support for undertaking all the investigations at the clinic.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e N/A\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest/Competing interests:\u003c/strong\u003e The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability:\u003c/strong\u003e Data is available and will be provided on the request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCode availability:\u003c/strong\u003e Code is available and will be provided on the request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions:\u003c/strong\u003e Ajeet was collected the data and entered the data. Ravi Datta Sharma, Abhimanyu Kumar Jha, Rakhi Singh, Rajeev Singh and Radhey Shyam Sharma were designed the manuscript. Ajeet, Shrawan K Trivedi, Ravi Datta Sharma and Radhey Shyam Sharma were analyzed the data and wrote the manuscript. Radhey Shyam Sharma, Ravi Datta Sharma, Rakhi Singh and Rajeev Singh were editing and supervised the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbalovich M et al. Hypothalamic-pituitary-testicular axis and seminal parameters in hyperthyroid males. Thyroid. 9(9):857-863 (1999).\u003c/li\u003e\n\u003cli\u003eAdah A S, Adah D I, Biobaku K T \u0026amp; Adeyemi A B. Effects of electromagnetic radiations on the male reproductive system. Anatomy Journal of Africa. Vol 7 (1): 1152 \u0026ndash; 1161 (2018). \u003c/li\u003e\n\u003cli\u003eAgarwal A, Deepinder F, Sharma R K, Ranga G \u0026amp; Li J. Effect of cell phone usage on semen analysis in men attending infertility clinic: an observational study. 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Infertility Fetal Med. 3:15-21 (2012).\u003c/li\u003e\n\u003cli\u003eVel\u0026aacute;zquez E M \u0026amp; Bellabarba G A. Effects of thyroid status on pituitary gonadotropin and testicular reserve in men. Arch Androl. 38(1):85-92 (1997).\u003c/li\u003e\n\u003cli\u003eVeldhuis J D. Male hypothalamic-pituitary-gonadal axis. In: Lipshultz LI, Howards SS, Ed. Infertility in the male, 3rd ed. Mooby-year Book St. Louis Mo. 23-58 (1997).\u003cstrong\u003e\u003c/strong\u003e\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"EMF, Semen Quality, RFR, Infertility, Reproduction","lastPublishedDoi":"10.21203/rs.3.rs-4162568/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4162568/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA prospective study has been conducted to find out the impact of cell phone on the semen quality and hormonal profile of the men. After fulfilling the exclusion and inclusion criteria 421 healthy male volunteer\u0026rsquo;s aged 18\u0026ndash;45 years were divided in to three groups depending on duration of cell phone use. Group-1 highly exposed group, Group-2 moderately exposed group, Group-3 control group. Subjects were asked to visit clinic every year for semen and blood sample collection up to three years. Semen physical characteristics and blood testosterone, T4 and TSH levels were estimated. Semen volume, rapid forward linear progression motility, moderate linear progression motility, sperm concentration, sperm density, percentage of normal and live sperm decreased in Group-1 in comparison to Groups-2 \u0026amp; 3 and trend continued till 3rd year. Sperm head abnormalities were higher in Group-1 than the Groups \u0026minus;\u0026thinsp;2 \u0026amp; 3 and trend continued till 3rd year. Slight decrease in levels of testosterone, significant increase in T4 and decrease in TSH were observed in Group-1 and similar trend was noticed till 3rd year. Present data indicate that the increased duration of cell phone use is adversely influencing the semen quality and hormone levels in men which may lead to infertility.\u003c/p\u003e","manuscriptTitle":"Impact of Radio Frequency Radiation (RFR) emitted from Cell Phone on Human Semen Quality","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-19 13:05:44","doi":"10.21203/rs.3.rs-4162568/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"3046c12d-43d7-41f9-b519-b8821a24e044","owner":[],"postedDate":"April 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-24T12:23:47+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-19 13:05:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4162568","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4162568","identity":"rs-4162568","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}