Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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On Sep 2018




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Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
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Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
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Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




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Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




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Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2022 | Month : October | Volume : 16 | Issue : 10 | Page : AC01 - AC05 Full Version

Histomorphometric Analysis of Chick Embryo Kidneys on Exposure to 1800 MHz and 2100 MHz Radiofrequency Radiation Emitted from Cell Phone


Published: October 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/58391.16862
Mary Hydrina Dsilva, Rijied Thompson Swer, Jayaraman Anbalagan

1. Associate Professor, Department of Anatomy, Zoram Medical College, Aizawl, Mizoram, India. 2. Professor and Head, Department of Anatomy, Zoram Medical College, Aizawl, Mizoram, India. 3. Ex-professor (Retired), Department of Anatomy, Mahatma Gandhi Medical College and Research Institute, Puducherry, India.

Correspondence Address :
Mary Hydrina Dsilva,
Associate Professor, Department of Anatomy, Zoram Medical College, Falkawn, Aizawl, Mizoram-796005, India.
E-mail: hydriswer@gmail.com

Abstract

Introduction: With its sophisticated and multifunctional features, the cell phone has become an integral part of human life. Scientific reports are still inconclusive, regarding the possible ill effects of Radiofrequency Radiation (RFR) emitted from cell phones on biological tissues.

Aim: To evaluate the possible tissue damage in developing kidneys of chick embryos, following exposure to 1800 MHz and 2100 MHz radiofrequency radiation emitted from 2G and 3G cell phones.

Materials and Methods: This experimental study was conducted in Department of Anatomy at Mahatma Gandhi Medical College and Research Institute, Puducherry, India, from August 2011 to June 2015. Fertilised chick embryos (144±20 eggs) were divided into three groups with a sample size of 48 eggs per group. Group A was exposed to 2G radiation (1800 MHz), group B was exposed to 3G radiation (2100 MHz) and group C was a sham exposed control group. The embryos were sacrificed from the 5th-12th day, and processed for routine histological procedures, to check structural and morphometric changes in the kidney. The standard epithelial height and nuclear diameter of both proximal convoluted tubule and distal convoluted tubule, karyorrhexis changes and diameter of urinary space were analysed using an ocular micrometer and square reticle. The results were statistically analysed using one-way Analysis of Variance (ANOVA).

Results: The results showed cytoplasmic changes (vacuolations) and nuclear changes (nucleomegaly, karyorrhexis) in proximal convoluted tubule and distal convoluted tubule, vascular changes (haemorrhage and infiltrations) in the interstitium and increased urinary space in the glomerulus of chick embryo kidneys.

Conclusion: Based on the study findings, it was concluded that RFR exposure from cell phones causes histopathological changes in the developing kidneys of chick embryos.

Keywords

Glomerulus, Interstitial oedema, Karyorrhexis, Oncosis, Standard epithelial height, Urinary space

Cell/mobile phones, a boon to the telecommunication industry, were first introduced in the form of analog cell phones in the 1980s. Since then, the cell phone industry has undergone tremendous growth with the introduction of 2G (900-1800 MHz), 3G (1900-2100 MHz), 4G (2300-2500 MHz) and recently rolled out 5G services (24 GHz-54 GHz) in as many as forty countries (1). With the enforcement of various online activities like telework, online classes, online payment, online shopping, online gaming, to curb the spread of Coronavirus Disease-2019 (COVID-19) pandemic by countries, the number of cell phone users has increased exponentially in the past two years. At present, there are 7.26 billion cell phone users in the world, which translates to 91.69% of the world’s population (current world population: 7.91 billion) (2).

Cell phones, while in use, emit non ionising Radiofrequency Radiations (RFR) which are absorbed into the user’s body causing both thermal and non thermal stress (3),(4). Initially, RFRs considered being harmless, but with the increasing scientific evidence of the ill effects of RFRs on human health and various animal models, it has become a global concern in the past decades. Studies have shown an increased mortality rate and various congenital anomalies in chick embryos on RFR exposure in the frequency range of 900-1800 MHz emitted from Global System Mobile communication (GSM/2G) cell phones (5),(6),(7). Exposure to a similar frequency also increased the rate of necrotic cells in blastocysts of mouse preimplantation embryos (8). Various authors have reported histopathological changes in the liver, kidneys, eyes, and brain of different animal models on exposure to RFR emitted from both second generation wireless telephony technology (2G) and third generation wireless telephony technology (3G) cell phones (9),(10),(11),(12),(13),(14). The role of RFR exposure ranging from 50-900 MHz causing altered antioxidant enzyme activities {Superoxide Dismutase (SOD), Catalase (CAT) and Glutathione Peroxidase (GPX)} in biological tissues of different animal models resulting in oxidative stress is well established (15),(16). Various in-vitro and in-vivo studies have also shown increased Deoxyribonucleic Acid (DNA) damage in the form of DNA strand breaks and rearrangement of DNA segments on biological tissues of both animal models and human beings on exposure to RFR ranging from 900-1800 MHz (17),(18).

Nevertheless, contradictory reports without any significant changes in the lens and retina of primate eyes on exposure to pulsed 60-1.25 GHz radiation and in the liver of mice models on exposure to 50 MHz electromagnetic fields, no effect on the level of antioxidants and nil DNA damage are also available (19),(20),(21),(22),(23),(24).

The controversies in the medical literature regarding the possible negative impacts of RFR emitted from cell phones on both human beings and animal models coupled with growing concerns about RFR exposure on health among the general public and various ethical issues involved in human research, have prompted us to take up the present study. Furthermore, the existing literature shows no comparative studies on histopathological changes in kidneys to RFR exposure of different frequency ranges from cell phones. Hence, the present study was conducted, to determine the effect of RFR exposure from 2G (1800 MHz) and 3G (2100 MHz) cell phone on chick embryo kidneys and to compare and conclude which frequency range is more detrimental to developing kidneys.

Material and Methods

This experimental study was conducted in Department of Anatomy at Mahatma Gandhi Medical College and Research Institute, Puducherry, India, from August 2011 to June 2015. The study followed all the procedures as per Ethical Guidelines for the care and use of experimental animals and was approved by Institutional Animal Ethical Committee (IAEC). Total (144±20) fresh fertile hen eggs Gallus gallus domesticus having similar weight (65-70±5 gms) were procured from Rajiv Gandhi College of Veterinary and Animal Sciences, Puducherry, India.

Inclusion and Exclusion criteria: Embryos that were healthy looking with a visible heartbeat and normal curvature were included in the study. Dead embryos, embryos showing congenital anomalies, and embryos without normal curvature were discarded.

The eggs were incubated in 12 batches of 12 eggs each, in a standard egg incubator (Standard digital incubator, Technico, India) at 37±0.5°C and 50-55% of humidity and ventilation. The fertilised hen eggs were grouped as A, B, and C with a sample size of 48 eggs per group.

• Group A was exposed to 2G cell phone radiation (900-1800 MHz)
• Group B was exposed to 3G cell phone radiation (1900-2100 MHz)
• Group C was treated as a sham exposed control group.

Study Procedure

The incubation of the eggs was carried out in a chamber fitted with a popular brand of cell phone {Specific Absorption Rate (SAR) of 0.310 watts/kilogram, power-2 watts}. The cell phone was hung at a distance of 5 cm from the eggs. For group C, the cell phone was kept in switched-off mode, whereas, for group A and B, the cell phone was kept in silent operative mode with the headphone plugged in, for its automatic activation while receiving a call. The service provider network was the same for both 2G and 3G exposure and the intensity of radiofrequency waves was measured using a radiofrequency meter (RF meter, Less EMF Inc. USA) that was kept inside the incubator.

The RFR exposure was initiated at the 12th hour of incubation by ringing from another cell phone. This was repeated every half an hour with the duration of the call being three minutes each. Thus, the total exposure for 12 hours was 72 minutes followed by 12 hours of exposure free period. The embryos were thus exposed to RFR up to the 12th day of incubation and six embryos per day were terminated from the 5th-12th day (9).

The chick embryos were fixed in 10% formaline and processed for routine histological study. A 5 μm thick sections were cut using a rotary microtome (Radical Senior Precision Rotary microtome, model: RMT- 30, India) from prepared paraffin blocks in the sagittal plane, coronal plane, and transverse plane and stained with Haematoxylin and Eosin (H&E) and Periodic Acid Schiff (PAS) stain (Merck, Germany).

Histomorphometric analysis: The following indices were observed of both Proximal Convoluted Tubule (PCT) and the Distal Convoluted Tubule (DCT)

• Standard Epithelial Height (SEH): The standard epithelial height of both PCT and DCT was measured from the basement membrane to the tubular lumen under 400x magnifications using a Trinocular research microscope fitted with microphotographic attachment (Olympus, model CH20i, Japan).
• Nuclear diameter: For the measurement of the nuclear diameter, healthy looking nuclei showing prominent nucleoli were measured under oil immersion.
• Diameter of urinary space of glomerulus: The outer diameter (A) and inner diameter (B) of the glomerulus were measured and the urinary space was calculated as A-B. All the measurement was taken from every third section of each slide using a calibrated ocular micrometer (10 mm scale: Erma, Model OM-010, India). Twenty random fields were selected from each embryo.
• Nuclear fragmentation (karyorrhexis): The number of nuclei showing karyorrhexis was counted in a randomly selected field using an eyepiece grid reticle of 0.5 mm squares containing 400 squares (Radical, India) under oil immersion from every third section of each slide (25). Randomly selected 50 fields were analysed for each embryo in control and experimental groups.

Statistical Analysis

The observed data were subjected to one-way Analysis of Variance (ANOVA) and the significance was determined using a Turkey’s post-hoc with p-value <0.05 for statistical significance. The statistical tests were done using the software Graph Pad Instat 3. All the data were expressed as Mean±SEM (Standard Error of the Mean).

Results

Histopathological changes: The kidneys of the control group embryos showed well-developed mesonephric tubules with normal structural features. The lining epithelial cells of both PCT and DCT showed intact basement membrane and brush border in PCT. The cells showed, fewer cytoplasmic vacuolations and relatively fewer nuclear changes in the form of nuclear fragmentation (karyorrhexis) and pyknosis (Table/Fig 1)a,(Table/Fig 2)a. The glomerulus appeared normal with intact cells lining the parietal and visceral layer, with a well-developed glomerular capillary network (Table/Fig 1)b,(Table/Fig 2)b.

However, 2G and 3G exposed embryos showed moderate to intense histopathological changes, in the form of increased vacuolations in the cytoplasm of lining cells, disruption of the luminal border and basement membrane of PCT. Cellular debris appeared in the lumen, due to the sloughing of lining cells (Table/Fig 1)c,e,(Table/Fig 2)c,e. The nuclei showed increased pyknosis and karyorrhexis in comparison with the control. The intensity of these changes increased as the age advanced. The cuboidal cells lining the DCT and collecting ducts appeared normal during the initial days of incubation. However, similar histopathological changes as that of PCT were observed in DCT in advanced age groups. Moreover, inflammatory changes in the interstitial spaces, diffused interstitial oedema with lymphocytes, plasma cells, monocytes, macrophages and haemorrhagic changes were also observed (Table/Fig 1)c,e,(Table/Fig 2)c,e. The glomerulus of both exposed groups showed degenerative changes with distorted glomerular capillary network and increased urinary space (Table/Fig 1)d,f,(Table/Fig 2)d,f.

Histomorphometric changes: The SEH of lining columnar cells of PCT showed a gradual increase as the age advanced in all three groups. The epithelial height was found to be significantly increased for both 2G group embryos (p-value <0.001) and 3G group embryos (p-value <0.001) when compared with control group embryos. On comparing 2G and 3G embryos, the 3G embryos showed increased height (p-value <0.01) (Table/Fig 3).

The SEH of lining cuboidal cells of DCT showed a gradual increase in height as the age advanced in all three groups. The epithelial height was found to be significantly increased in 2G (p-value <0.001) and 3G group embryos in comparison with the control group (p-value <0.001). On comparing 2G and 3G embryos, the 2G embryos showed increased height that was significant on the 5th, 6th, and 7th day (p-value <0.05, p-value <0.05, and p-value <0.001 respectively) and the 3G group embryos showed significantly increased height on the 12th day of incubation (p-value <0.001) (Table/Fig 4).

The nuclear diameter of PCT increased gradually in all three groups with advancing age. The diameter was significantly increased in both the 2G and 3G exposed groups than in the control group (p-value <0.001). In the 3G group, embryos showed an increased diameter that was more significant (p-value <0.05) than in the 2G group (Table/Fig 5).

The nuclear diameter of DCT of all three groups showed a gradual increase in diameter as the age advanced. The 2G embryos showed an increased diameter than the control group embryos (p-value <0.05). The 3G group also showed an increased nuclear diameter on comparing with the control group and the increase was significant only on the 5th and 6th days (p-value <0.05). On comparing 2G and 3G groups, the 2G group embryos, showed a significant increase in diameter (p-value <0.05) (Table/Fig 6).

The urinary space showed a gradual increase in diameter as age advanced in all three groups. Both the 2G and 3G groups showed a statistically significant increase in the urinary space on all days as compared with the control group embryos (p-value <0.001 and p-value <0.001 respectively). On comparing 2G and 3G groups, 3G group embryos showed an increased diameter of urinary space that was significant on all days (p-value <0.05) (Table/Fig 7).

In both 2G and 3G group, embryos showed increased karyorrhexis than the control group embryos (p-value <0.05 and p-value <0.01 respectively). On comparing between 2G and 3G group, the 3G group showed increased karyorrhexis than the 2G group, however, the increase was significant only on the 5th, 6th, and 12th day (p-value <0.001) (Table/Fig 8).

Discussion

Kidneys are moderately sensitive to radiation. The effects of radiations were observed in convoluted tubules and glomerulus in the form of cytoplasmic changes, nuclear changes, and vascular changes (26).

The histopathological changes observed in the PCT and DCT of the present study were similar to the observations made by researchers in mice exposed to RFR (900 MHz) from the mobile phone for variable exposure duration (27),(28),(29). Al-Glaib B et al., (2008), reported glomerular atrophy, interstitial infiltrations, and vacuolations in renal tubules by exposing mice to 900 MHz RFR for a duration of one hour per day for 10 days (27). A similar study by Hanafy LK et al., also reported glomerular atrophy and extravasated blood cells between renal tubules on RFR exposure of one hour per day for four weeks in rat models (28). Khayyat LI exposed mice for a longer duration of eight hours for three days and 12 days and their findings showed atrophied glomeruli, cytoplasmic vacuolations and pyknosis of lining epithelial cells of the renal tubules, renal vein congestion and inflammatory changes between the renal tubules (29). Ingole IV and Ghosh SK, reported similar changes in PCT with narrowed bowman’s space of renal corpuscles in chick embryos on exposure to RFR of 900 MHz for four hours, five hours, and six hours for six, eight and 10 days (11).

In the present study, DCT showed isolated vacuolations; in PCT, the vacuolations were present in almost entire cells lining the tubules indicating necrotic changes. The vacuolations in the cytoplasm might be due to fatty metamorphosis or steatosis as they were PAS negative (Table/Fig 2)c,e (30).

The oedema and presence of RBCs in the interstitium of kidney tissue observed might be due to vascular damage caused by RFR. Exposure to RFR causes vascular dilatation, and vacuolations of the endothelium of peritubular capillaries with focal necrosis, resulting in haemorrhage and exudation of blood plasma containing variable infiltrates of lymphocytes, plasma cells, monocytes and macrophages. These changes were similar to the changes observed in acute interstitial nephritis (26),(31). Interstitial oedema in the kidney tissue after RFR exposure was also reported by Al-Glaib B et al., Hanafy LK et al., and Khayyat LI (27),(28),(29).

The radiations are known to cause nuclear swelling (nucleomegaly), mild to moderate pyknosis, karyorrhexis (nuclear fragmentation), and karyolysis (dissolution of chromatin) (26). In the present study, karyorrhexis and changes in nuclear diameter were significantly increased in 3G group embryos due to the lethality of 3G radiation. Another observation was the increase in SEH of lining cells in PCT and DCT of both 2G and 3G group embryos as compared with control group embryos. RFR emitted from cell phones caused degeneration or weakening of plasma membrane due to non thermal effects (32),(33). This would have resulted in an increased influx of water (hydropic changes/oncosis) resulting in cellular swelling (cytomegaly) (26),(30). The 3G group embryos showed significantly increased SEH in PCT and on the 12th day in DCT. This probably might be due to the increased lethality of RFR emitted from the 3G cell phone.

The increased glomerular diameter observed, might be due to cytoplasmic changes caused by RFR. Exposure to RFR would have resulted in hydropic changes causing cytomegaly of cells, lining the parietal and visceral layer of glomerulus and endothelial cells lining the glomerular capillaries, which in turn resulted in an increase in glomerular diameter [26,30]. Moreover, the endothelial cells lining the blood vessels are highly sensitive to radiation effects and it manifests as vasodilatation and increased vascular permeability. In present study, these changes in glomerular capillaries, caused the plasma exudate to escape into the urinary space resulting in increased urinary space [26,31]. This observation was similar to the findings of Mugunthan N et al., who reported dilated glomeruli and urinary space in mice kidneys exposed to RFR of 900-1900 MHz for 48 minutes per day for a period of 30-180 days (12). However, few studies reported glomerular atrophy on RFR exposure in rat and mice models as the duration of exposure was 3-12 days (27),(28),(29),(34).

Hence, the effect of RFR exposure manifests differently, depending on the duration of exposure, frequency, and intensity of transmission, the number of exposures, the shape and size of the exposed organism, the water and mineral content of the organism, and the distance from the radiation source (35),(36).

Limitation(s)

Direct extrapolation of results of the present study on chick embryos to the human population may be limited, as the volume and size of kidneys, the architecture of renal tissue, tissue electromagnetic properties, penetration and interaction of RFR into the body might vary from one species to another. The present study was done only from 5th-12th day (organogenesis period) of incubation. Further study on prolonged exposure (growth and development period) is recommended.

Conclusion

Based on our experimental outcome, we conclude that the chronic exposure of chick embryo kidneys to RFR emitted from 2G and 3G cell phones resulted in histopathological and histomorphometric changes in kidney structures. Both the exposed groups showed increased cytoplasmic vacuolations, disrupted luminal border and the basement membrane, increased epithelial height and nuclear diameter, increased pyknosis and karyorrhexis in PCT and DCT as well as interstitial oedema with infiltrations, and increased urinary space of glomerulus. The 3G group embryos showed significantly increased changes because of the lethality of 3G radiations.

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DOI and Others

DOI: 10.7860/JCDR/2022/58391.16862

Date of Submission: Jun 11, 2022
Date of Peer Review: Jul 21, 2022
Date of Acceptance: Aug 27, 2022
Date of Publishing: Oct 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? NA
• For any images presented appropriate consent has been obtained from the subjects. NA

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