{"id":61254,"date":"2024-09-30T11:18:25","date_gmt":"2024-09-30T11:18:25","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=61254"},"modified":"2024-10-11T16:12:27","modified_gmt":"2024-10-11T16:12:27","slug":"the-impact-of-mobile-phone-electromagnetic-waves-on-the-neurons-and-blood-brain-barrier-integrity-in-the-chick-embryo","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no3\/the-impact-of-mobile-phone-electromagnetic-waves-on-the-neurons-and-blood-brain-barrier-integrity-in-the-chick-embryo\/","title":{"rendered":"The Impact of Mobile Phone Electromagnetic Waves on the Neurons and Blood Brain Barrier Integrity in the Chick Embryo"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Mobile phones utilize non-ionizing low\nelectromagnetic waves (EW), causing rapid environmental pollution due to\nexposure to radio waves at frequencies of 800-900 Mega Hz. Excessive exposure\nto these EW may increase the production of Radical Oxygen Species (ROS). Excessive\nuse of mobile phones by teenagers and pregnant mothers raises concerns due to\ntheir increased radiosensitivity.  Teenagers\ndeveloping bodies and the rapid division of embryonic cells in fetuses\npotentially heighten their vulnerability to electromagnetic waves. Cell phone\nexposure to prenatal and postnatal children resulted in behavioral problems 1.<\/sup>\nThese electromagnetic waves affect the brain and memory 2<\/sup>. Xu\ndemonstrated mt DNA and levels of mitochondrial RNA in the neuron was affected\nby the electromagnetic fields 3-4<\/sup>. High levels of electromagnetic\nwaves in mother\u2019s bedroom during pregnancy may cause autism in the baby 5<\/sup>.  <\/p>\n\n\n\n

The effect\nof radio waves in the adult brain was also a point of interest for\ninvestigators due to its proximity to the brain and ears while talking on\nmobile phones. Reports mention increased headache, fatigue, irritability,\nstress, sleepiness, concentration difficulties, nausea, lack of appetite,\nblurred vision, depression 6-12<\/sup>. Disturbances in sleep and\nalteration of synaptic plasticity, neurotransmitter release and life cycle of\nnerve cells because of the use of cellular phone 13-14<\/sup>.\nNeuropsychiatric changes such as depression, somatization, obsessive\ncompulsivity, phobic anxiety, paranoid ideation, sleeping disturbances were\nalso reported in literature 14-18<\/sup>. The increasing incidence of\ndepression and increasing use of mobile phones seems to directly relate to each\nother suggests memory disfunction, attention dysfunction, decreased motor\nfunction, decelerated reaction time and lowered neuromuscular strength 19<\/sup>.  Asad reported a positive relationship between\nanxiety and depression with smartphone addiction in South Korea 20<\/sup>.\nIn another study, Siddiqi also mentioned the excessive use of smartphones by\nmedical students 21<\/sup>.  <\/p>\n\n\n\n

Salford\nreported that prenatal and postnatal exposure of radio waves stimulate neural\ncell death and prevents stem cells to change into adult neurons 4<\/sup>.\nIt is now understood that many neurodegenerative diseases such as Alzheimer\u2019s\ndisease, Parkinson\u2019s disease and amyotrophic lateral sclerosis may be due to damage\nto mitochondria of the neurons 22-28<\/sup>. Furthermore, Down syndrome 29-30<\/sup>,\nHuntington\u2019s disease 31<\/sup>, Friedreich ataxia; oxidative damage of\nmitochondria is most likely playing a major role in the pathogenesis of these\ndiseases 32-34<\/sup>.<\/p>\n\n\n\n

Not much research has been done on the negative side\neffects of EW on the nervous system. The study revealed more profound knowledge\non the effects of EW exposure on the neurons, neuroglia, blood brain barrier\nand brain intercellular matric. <\/p>\n\n\n\n

Hypothesis<\/strong><\/p>\n\n\n\n

The radio waves affect the developing neurons and the neuroglia in the developing chick embryo.   <\/p>\n\n\n\n

Objectives<\/strong><\/p>\n\n\n\n

To expose the developing chick embryo with EW for 10 and 15 days starting from day zero.<\/p>\n\n\n\n

To examine the morphological changes in the cerebral cortex and cerebellum at days 10 and 15 of embryo development in the control and exposed groups.<\/p>\n\n\n\n

Observe the blood brain barrier in the exposed and control groups. <\/p>\n\n\n\n

Materials and Methods <\/strong><\/p>\n\n\n\n

Chicken fertilized eggs (\u2018Cobb\u2019 Gallus gallus\ndomesticus) <\/em>were received from Sohar poultry. This animal model\nhas been used extensively for research purposes 35<\/sup>.<\/strong><\/p>\n\n\n\n

An incubator\n(Model EH-35, Sino-PFE Company, China)\nwith a capacity of holding 30 eggs at a time and with computerized control of temperature,\nhumidity and air ventilation was utilized (Fig.1). Eggs were rotated ten times per\nday.  Forty\nzero-day fertilized eggs were divided equally at random into control and exposed groups. In each group, 20 eggs were\nplaced inside the incubator and the temperature was set to 37 degrees and humidity\n50-60% by the control panel.  <\/p>\n\n\n\n

In the exposed groups, the eggs were subjected to EW released by a cell phone placed inside the incubator, while in the control group the eggs were unexposed to EW. A specific branded cell phone with a common cellular service provider was used, operating at 1800 MHz, with a power output of 0.47 W\/kg and SAR of 1.10 W\/kg (head). The intensity of the EW during the experiment was measured using a Tri Field Meter, model 100XE. The cell phone was activated for 5 minutes at a time, ten times daily, by calling from another cell phone outside the incubator, with no exposure periods in between. Calls were not made during the night. This resulted in a total daily exposure duration of 50 minutes, starting from day zero. The chick embryos were sacrificed on day 10 (500 minutes of exposure) and day 15 (750 minutes of exposure). The egg fully develops in 21 days, day 10 and 15 of development is a reasonably good interval to observe the effects on cells and had been used in other studies35<\/sup>. The eggshell was carefully served by scissors, the membranes were removed around the embryo, the chest wall then opened with sharp micro scissors, and the heartbeat was observed. The brain of the embryo was dissected and removed after opening the bones of the skull. The cerebral cortex and cerebellum of the brain were dissected, and specimens were removed for fixed for EM studies (Fig.2).  <\/p>\n\n\n\n\n\n
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Figure 1: A 30-egg incubator<\/strong><\/p>\n

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Figure 2: Day 10 a) Removal of the embryo on day 10 b) brain at day 10. c) Opening of the skull of chick embryo on day 15 d) brain at 15.<\/strong><\/p>\n

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Control Group<\/strong><\/p>\n\n\n\n

20 eggs were incubated under the same environment except that the cell phone was placed without a battery to make sure that there is no EW emitted. The embryos were dissected at day 10 and 15, brain was removed and specimens of cerebral cortex and the cerebellum were removed and fixed for EM study. <\/p>\n\n\n\n

Electron microscopy (EM)<\/strong><\/p>\n\n\n\n

Brain specimens were fixed in glutaraldehyde for 5 hours, followed by washing in buffer solution. Further processing for the EM was done at EM Laboratory at Sultan Qaboos University, Oman. Same number of specimens of cerebral cortex and cerebellum in both the groups were taken to the EM lab for processing. Processing of the samples were done based on a protocol by Bozzola and Russell with a few changes. All samples were immediately placed on vials containing EM fixative (2.5% Glutaraldehyde) and left for 2 hours. Tissues were then washed twice using cacodylate buffer (PH 7.2-7.4) 10 minutes each, post fixation was carried using osmium tetroxide for one hour and dehydration was done using series of acetone starting from washing with distilled water, 25% acetone, 75% acetone, 95% acetone and 99.9% acetone. Every step was carried for 10 minutes. Tissues then were placed in 1:1 acetone to pure epoxy resin for one hour then in 1:3 acetone to pure epoxy resin for half an hour. Next it was placed in pure epoxy resin for one hour followed by fresh pure resin for half an hour. Embedding was carried out in size 00 embedding beam capsules (Agar) and polymerized in 70\u00b0C oven for 12 hours. <\/p>\n\n\n\n

All tissue blocks were cut using Lieca\nultramicrotome for semi thin sectioning (0.5 um) with glass knives to allocate\nthe areas of interest using light microscopy then were again cut to have\nultrathin sections (60-90 nm thick) with diamond knives and picked up on 300\nmesh cupper grids. All sections were stained using uranyl acetate and lead\ncitrate. Sections were then observed using JEOL JEM-1230 EX Japan transmission\nelectron microscope and EM photos were subsequently analyzed.<\/p>\n\n\n\n

Results<\/strong><\/p>\n\n\n\n

Brain: 1) TEM of Cerebral cortex: Control group<\/strong><\/p>\n\n\n\n

Day 10: Cerebral cortex of the chick embryo showing large well-developed neurons (pyramidal neurons and oligodendrocytes). The nerve fibers can be seen occupying the space in between the neurons; however, many empty spaces could be seen in between the neurons (Fig.3a.b).   <\/p>\n\n\n\n\n\n
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Figure 3: Control group at Day 10: a) Transmission electron microscope (TEM) analysis shows multiple well developed healthy large pyramidal neurons (black arrow), neuroglial cells (red arrow) and thin extracellular matric (ECM).<\/strong><\/p>\n

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Day 15: Cerebral cortex of the chick embryo shows large well-developed neurons (pyramidal neurons) and oligodendrocytes and clear nucleus. The nerve fibers are well developed and organized regularly with small gaps in between. (Fig.4 a, b). <\/p>\n\n\n\n\n\n
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Figure 4: Control group on Day 15: 4a) Transmission electron microscope (TEM) analysis shows many well-developed pyramidal neurons (arrow), nerve fibers and neuroglia and in the cerebral cortex.<\/strong><\/p>\n

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Brain: 2) TEM of Cerebral cortex: Exposed group<\/strong> <\/p>\n\n\n\n

Day 10: The neurons showing changes of apoptosis, chromatin condensation, cellular membrane not visible as compared to the control group. Neurons become shrunken and appear smaller than healthy neurons, with condensed cytoplasm and reduced organelle density. The loss of cytoplasmic volume is often accompanied by a distorted cellular shape and irregular contours. The extracellular matrix (ECM) becomes dense and dark and much fewer white gaps visible thus filling most of the matrix. Axons in the matrix are not clearly seen (Fig.5a).<\/p>\n\n\n\n

Day 15: Shrunken and atrophied neurons with chromatin condensation and degenerative alteration were observed in the pyramidal neurons; nucleus membrane blebbing was also apparent. The extracellular matrix (ECM) became very dense and disorganized when compared to the control group.  (Fig 5b,c).  <\/p>\n\n\n\n\n\n
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Figure 5: Exposed day 10 and day 15: a) Transmission electron microscope (TEM) analysis on day 10 shows multiple cortical neurons whose nucleus are not clearly visible and dense extracellular matrix as compared to the control.<\/strong><\/p>\n

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Brain: 3) TEM of Cerebellum: Control<\/strong><\/p>\n\n\n\n

Day 10: cerebellum shows multiple large neurons and patchy extracellular matrix with nerve fibres. These large neurons are most likely Purkinje cells. The extracellular matrix is less dense with empty spaces in between the neurons. (Fig.6a). <\/p>\n\n\n\n

Day 15: Multiple large neurons with clear margins and prominent nucleoli can be seen. The extracellular matrix density is increased than day 10 (Fig 6b). The mitochondria in the neurons were oval or slender in shape (Fig.8a). A normal blood brain barrier can be seen with tight junctions between the capillary endothelia (Fig.9a.).<\/p>\n\n\n\n\n\n
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Figure 6: Cerebellum: Control: a) Transmission electron microscope (TEM) analysis of control group on
day 10 shows few large cerebellar (Purkinje) neurons (arrow) and light extracellular matrix containing
nerve fibers (yellow arrow) in the less dense extracellular matrix.<\/strong><\/p>\n

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Figure 7: Cerebellum: Exposed: a) Transmission electron microscope (TEM) analysis on day 10 shows an increased number of Purkinje neurons (arrow) with rounded nucleus, dense extracellular matrix (yellow arrow) with the nerve fibers and as compared to control.<\/strong><\/p>\n

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Brain: 4) TEM of Cerebellum: exposed group<\/strong><\/p>\n\n\n\n

Day 10: Multiple large neurons were seen and nucleus showing chromatin condensation. The extracellular matrix is much denser as compared to the control and many black apoptotic granules in the extracellular matrix were observed (Fig 7a).  <\/p>\n\n\n\n

Day 15: The neurons were shrunk, and chromatin condensation was apparent. The neurons show multiple rounded swollen mitochondria. (Fig.7b, 8b) At higher magnification the mitochondria can be seen swollen and irregular in shape and rupture of the inner and outer membranes was noted (Fig 8c).  A blood brain barrier shows disruption of the tight junctions and damage of capillary endothelium (Fig.9b.).<\/p>\n\n\n\n\n\n
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Figure 8: Cerebellum: Exposed group Day 15: a) Transmission electron microscope (TEM) analysis in the control group on day 15 shows multiple elongated or oval shaped mitochondria (arrow) with cisterns present in the neurons (arrow) TEM original magnification X 40000.<\/strong><\/p>\n

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Figure 9: a) The capillary shows well developed endothelial cell (green arrow) with tight junction (black arrow) and pericyte (yellow arrow) forming the blood brain barrier. The foot process of astrocytes can also be seen surrounding the capillary (orange arrow).<\/strong><\/p>\n

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Figure 10: The diagram showing the mechanism of how the ORS can lead to mitochondrial swelling and cell death.<\/strong><\/p>\n

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Discussion<\/strong><\/p>\n\n\n\n

EW adverse effects were studied in the\nchick embryo brain tissues on days 10 and 15 of its fetal development. The\neffects of the electromagnetic waves are classified as thermal and non-thermal.\nThermal effects are now controlled by strict regulation on Specific Absorption\nRate (SAR); this is a measure of the rate at which energy is absorbed per unit\nmass by the body when exposed to EW. Regulatory bodies like the FCC set limits\non SAR to ensure safety (e.g., 1.6 W\/kg in the U.S.). Non-thermal effects such\nas oxidative stress, damage to mitochondria, cell wall, DNA etc. need further\nresearch. This study explores the effects of EM on the neurons, neuroglia and extracellular\nmatrix in a developing chick embryo.  In\nthe control groups, neurons were well developed surrounded by oligodendroglia\ncells and the nerve fibers were showing increasing density from days 10 to 15.\nIn the exposed groups, the neurons showed shrunken morphology with degenerative\nalteration, nucleus not clearly visible, extracellular matrix became very dense\nand disorganized as compared to the control. Oligodendrocytes were also not\nclearly visible. <\/p>\n\n\n\n

Sadequl\nIslam reported darkly stained nuclei in degenerated neurons and decrease in healthy\nneurons when the chick embryo was exposed with electromagnetic waves 36<\/sup>.\nOn day 14 of chick development, the number of degenerated neurons with\nhyperchromatic deeply stained nucleus were significantly more in the group exposed\nthan the healthy neurons in the control group (p=0.05). <\/p>\n\n\n\n

Hasan 37<\/sup>\nobserved similar results and found fewer neurons in the exposed embryo.\nApoptosis in a few neurons were also observed in the exposed group 38-41<\/sup>.\nRadiation causes an increase in superoxide dismutase, catalase and\nmalondialdehyde levels in the embryo.   <\/p>\n\n\n\n

Eser\nreported that radio waves caused morphological alterations in the cerebral cortex,\nbrain stem and cerebellum in albino male rats 42<\/sup>. They observed marked\ndegenerative changes, decrease in cytoplasm and dark pyknotic nuclei and a\nsmaller number of neurons in the EM exposed group versus the control group. <\/p>\n\n\n\n

Further\nreported in literature that radio waves of 900 MHz have caused pyknotic neurons\nin the hippocampus, deeply stained granules in cerebellum 43-45<\/sup>.  This study also observed deeply stained\ngranules in the cerebellum which are stress granules (SRs). These stress\ngranules inhibit stress-induced apoptosis and help the cell to repair. <\/p>\n\n\n\n

Romeo did a\nmeta-analysis on the fact that electromagnetic waves exposure causes apoptosis\nand found a mixed result; however, many studied reported apoptosis as a result\nof electromagnetic exposure 46<\/sup>.<\/p>\n\n\n\n

Islam also\nreported an increase of VEGF-A expression in the exposed group which mean\ninflammation and hypoxia leading to oxidative stress induced by the radio waves\n36<\/sup>. Vahid  reported the\nelectromagnetic radiation induced the entry of calcium via TEPV1 channel in\nhippocampus and dorsal root ganglia in rats resulting in apoptosis 47<\/sup>.  Absence of mitochondrial function, rise in\namyloid beta expression and stimulation of apoptotic factors e.g. caspase-9 and\n-3 in the hippocampus after EMR-2450 MHz exposure was reported 48<\/sup>.<\/p>\n\n\n\n

Blood brain\nbarrier is another structure which can be affected by electromagnetic waves.\nPersson  reported that radiation from\ncell phones can damage the blood brain barrier in rats allowing albumin to\nenter the brain 49<\/sup>.  This\nstudy observed very dense extracellular matrix due to which the nerve fibers\ncannot be identified. It seems that rupture of the blood brain barrier in the\nexposed group on day 15, resulted in leakage of albumin in the extracellular\nmatrix giving it a dense matrix appearance. (Fig.7a, b). <\/p>\n\n\n\n

Salford, L.\nG investigated the impact of microwave exposure from GSM mobile phones on rat brains,\nspecifically looking at the blood-brain barrier and neuron damage and reported\nsignificant damage of the neurons of the cortex, hippocampus and basal ganglia\nbecause of albumin leakage through blood-brain barrier 50<\/sup>. Stam\nreported damage of blood brain barrier by radio waves exposure in animal model\nhowever stated that in humans the evidence of damage is still not reported 51<\/sup>.\nLeszczynski   in his research focused on\nhow mobile phone radiation can non-thermally activate hsp27 stress pathways in\nhuman endothelial cells, thereby increasing permeability of blood-brain barrier\n52<\/sup>. Nittby also reported the permeability changes in blood-brain\nbarrier of rats after exposure to GSM-900 mobile phone radiation and found\nsignificant correlation of albumin excavation and exposure level 53<\/sup>.\nEberhardt, J. L. in his research provides an overview of how microwave\nradiation affects blood-brain barrier permeability, leakage of albumin into the\nextracellular matrix and resulting in neuron damage in rats54<\/sup>. Shabani\nalso reported increased permeability of the blood-brain barrier in rats55<\/sup>.\nGao examined albumin immunohistochemistry and Evans blue staining after\nexposure of rat\u2019s brain to electromagnetic pulses. Zonula occludens were\nevaluated using western blotting, results revealed increased permeability of\nblood brain barrier 56<\/sup>. This research revealed that the blood brain\nbarrier in the cerebellum was damaged, and leakage of albumin was evident in\nthe extracellular matrix of the exposed group. <\/p>\n\n\n\n

This study\nalso reported the degenerative changes in the cerebral cortical and cerebellum\nneurons after exposure to radio waves. This exposure also affected the\nmitochondria in the cerebellum neurons and observed deformity of the mitochondria\nwhich became rounded instead of oval or elongated. This agrees with other\nreports mentioned in literature. Mitochondrial DNA damage due to oxidative stress\nin primary cultured neurons were observed 3<\/sup>. Increased permeability to\ncalcium due to ROS production leads to mitochondrial swelling. Vicious cycle causing\nfurther increase in ROS triggers apoptosis (Fig.10). First sign of mitochondria\ncell membrane damage is its swelling, which increase with further increase\nproduction of ROS, and the mitochondria will become round. Oxidative\nstress cause mitochondrial DNA mutation, damage mitochondrial respiratory\nchain, alter membrane permeability and effect ca+ homeostasis and mitochondrial\ndefense system 57<\/sup>. Mitochondria became swollen and vacuolated in\nmotor neurons in mice 58<\/sup>. In this study, mitochondria were observed\nto be swollen in the cerebellum in the exposed group versus the control where\nthey were oval and elongated. Permeability of mitochondrial membrane depends on\ncommunication between ca2+<\/sup> and ROS system. A major cause of\nEM-induced calcium influx into the mitochondria is increased sympathetic\nactivity 59<\/sup>. ROS when stimulated will produce free O +2 on the mitochondrial\ninner surface. This free O+ 2 will invade thiol protein to open up the transition\npores, increases membrane permeability and causes mitochondria to swell 60<\/sup>.\nMitochondria is the most sensitive organelles to oxidative stress and swells under\nsuch conditions 61<\/sup>. <\/p>\n\n\n\n

To carry\nout highly specialized functions the neurons require more energy as compared to\nother cells. Mitochondria generate energy, and its dysfunction thus plays a\nmajor role in different neurodegenerative diseases. It is now understood that abnormal\nmitochondrial dynamics pertinent to neuronal synaptic loss and cell death may\nbe a cause of Alzheimer’s disease, Parkinson’s disease, and Huntington’s\ndisease 48<\/sup>. Abnormal mitochondrial morphology was reported in mutants\nof Parkinson’s disease-related genes.<\/p>\n\n\n\n

Mitochondria\nare unique organelles within cells, distinct from the nucleus, as they contain\ntheir own DNA known as mitochondrial DNA. They are capable of synthesizing\ntheir own RNA and proteins, and has the mitochondrial respiratory chain on its\ninner membrane 62<\/sup>. This chain comprises five complexes\u2014namely I, II,\nIII, IV, and V. The first four complexes are accountable for converting\nadenosine diphosphate (ADP) into adenosine triphosphate (ATP)63<\/sup>.<\/p>\n\n\n\n

1-5% oxygen is\nconverted into ROS under normal conditions; mitochondria is primary source of\nintracellular ROS 64<\/sup>. Complex III is the major site of ROS\nproduction under typical metabolic conditions 65<\/sup>. Elevated ROS\nlevels can damage these complexes and other mitochondrial macromolecules,\nincluding lipids, proteins, and DNA 66<\/sup>. Damage to mitochondrial DNA\nexacerbates oxidative stress, creating a harmful cycle of ROS generation and\neventually leading to apoptosis, which can disrupt mitochondrial energy\nproduction 67,68<\/sup>.<\/p>\n\n\n\n

Increased\npermeability of the mitochondrial inner membrane disrupts mitochondrial calcium\nhomeostasis 60<\/sup>. Elevated calcium levels enhance the production of\nsuperoxide radicals, propagating a cycle of damage. Excessive calcium\naccumulation leads to osmotic swelling and damage of the outer mitochondrial\nmembrane 69<\/sup>. Increased ROS production further compromises membrane integrity,\nboosts calcium uptake, and triggers apoptosis and cell death 46, 70<\/sup>.<\/p>\n\n\n\n

Calcium ions\nmust enter the neuron’s cytosol for neurotransmitter release 5<\/sup>.\nNeuronal signaling relies on the release of neurotransmitters, which is\ninitiated by a brief influx of calcium into the cytosol. Exposure to radio\nwaves can cause membrane leaks, elevating intracellular calcium concentrations.\nThis heightened calcium level causes the cell to be overly responsive,\nreleasing more neurotransmitters and leading to increased brain activity 71-74<\/sup>.\nConsequently, the brain can become overloaded with excessive signals,\npotentially resulting in concentration issues and attention deficit\nhyperactivity disorder (ADHD).<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

We concluded that electromagnetic waves have produced damage to the cerebral and cerebellar neurons and shown increased density in the extracellular matrix of the developing chick embryo because of damage to the blood-brain barrier. Brain blood-barrier damage and leakage of albumin are the most probable causes of the increased density in the extracellular matrix and neuronal damage. Swelling of the mitochondria and damage to its membrane might be due to an increase in oxidative stress caused by electromagnetic waves. To bridge our findings with broader health implications and an increase in the neurodegenerative disease in old age, further research is needed to understand how the observed cellular changes found in this study due to exposure of electromagnetic waves could translate into the development of neuro degenerative diseases.<\/p>\n\n\n\n

Acknowledgements<\/strong><\/p>\n\n\n\n

Deeply acknowledge the Department of Pathology at SQU for processing the specimens for EM and allowing the use of the electron microscope. I am also thankful to our Dean, Prof. Mohammed Al Shafaee, whose tremendous encouragement is always stimulating us to continue research and allow us to visit SQU.  11, 16<\/p>\n\n\n\n

Conflicts of Interest<\/strong><\/p>\n\n\n\n

The author(s) do not have any conflict of interest <\/p>\n\n\n\n

Funding<\/strong> Sources<\/strong><\/p>\n\n\n\n

The study was supported by Oman Ministry of Higher Education (RG) grant no: MOHERI\/BFP\/RGP\/18\/161 <\/p>\n\n\n\n

Data Availability<\/strong> Statement<\/strong> <\/p>\n\n\n\n

<\/strong>The authors confirm that the data supporting the findings of this study are available within the article and with the main author. The raw data that support this study is available at the EM laboratory at Sultan Qaboos University, Oman, which are available from the 1st<\/sup> author upon reasonable request.   <\/p>\n\n\n\n

Ethics Statement<\/strong><\/p>\n\n\n\n

This article was approved by the research committee of the college of medicine and health sciences, National university of science and technology<\/p>\n\n\n\n

Informed Consent Statement<\/strong><\/p>\n\n\n\n

This study did not involve human participants, and therefore, informed consent was not required<\/p>\n\n\n\n

Author\u2019s contribution<\/strong><\/p>\n\n\n\n

Prof. Najam Siddiqi: Writing the proposal, doing the experiment, collecting the data, analyzing the data, writing the paper<\/p>\n\n\n\n

Dr Faisal Moin: Analyzing the data, writing the paper, collection the references, and putting them in order<\/p>\n\n\n\n

Mr. Mohammed Al Kindi: Processing the EM mesh for EM, Taking the pictures and analyzing the data<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

  1. Divan HA, Kheifets L, Obel C, Olsen J. Prenatal\nand postnatal exposure to cell phone use and behavioral problems in children.\nEpidemiology. 2008 1;19(6):S94-5.<\/li>
  2. Chaturvedi CM, Singh VP, Singh P, Basu P,\nSingaravel M, Shukla RK, Dhawan A, Pati AK, Gangwar RK, Singh S. 2.45 GHz (CW)\nmicrowave irradiation alters circadian organization, spatial memory, DNA\nstructure in the brain cells and blood cell counts of male mice, mus musculus.\nProgress In Electromagnetics Research B. 2011; 29:23-42.<\/li>
  3. Xu S, Zhou Z, Zhang L, Yu Z, Zhang W, Wang Y,\nWang X, Li M, Chen Y, Chen C, He M. Exposure to 1800 MHz radiofrequency\nradiation induces oxidative damage to mitochondrial DNA in primary cultured\nneurons. Brain research. 2010 22; 1311: 189-96.<\/li>
  4. Salford LG, Brun AE, Eberhardt JL, Malmgren L,\nPersson BR. Nerve cell damage in mammalian brain after exposure to microwaves\nfrom GSM mobile phones. Environmental health perspectives. 2003;111(7):881-3.<\/li>
  5. Klinghardt,\nDietrich, MD. Pilot Study. Does high radiation exposure during pregnancy cause\nautism. 2018:\nhttps:\/\/www.dqindia.com\/high-radiation-exposure-pregnancy-cause-autism\/ <\/li>
  6. Frey AH. Headaches from cellular telephones:\nare they real and what are the implications? Environmental health perspectives.\n1998;106(3):101-3.<\/li>
  7. Huber R, Treyer V, Borbely AA, Schuderer J,\nGottselig JM, Landolt HP, Werth E, Berthold T, Kuster N, Buck A, Achermann P.\nElectromagnetic fields, such as those from mobile phones, alter regional\ncerebral blood flow and sleep and waking EEG. Journal of sleep research.\n2002;11(4):289-95.<\/li>
  8. Bortkiewicz A, Zmy\u015blony M, Szyjkowska A,\nGadzicka E. Subjective symptoms reported by people living in the vicinity of\ncellular phone base stations: review. Medycyna pracy. 2004  1;55(4):345-51.<\/li>
  9. Chu MK, Song HG, Kim C, Lee BC. Clinical\nfeatures of headache associated with mobile phone use: a cross-sectional study\nin university students. BMC neurology. 2011;11:1-7.<\/li>
  10. Dasdag S, Akdag MZ, Ulukaya E, Uzunlar AK, Ocak\nAR. Effect of mobile phone exposure on apoptotic glial cells and status of\noxidative stress in rat brain. Electromagnetic biology and medicine. 2009 1;28(4):342-54.<\/li>
  11. Dwyer MJ, Leeper DB. A Current Literature\nReport on the Carcinogenic Properties of Ionizing and Non-: Ionizing Radiation.\n1978.<\/li>
  12. Salama OE, Abou El Naga RM. Cellular phones:\nare they detrimental? The Journal of the Egyptian Public Health Association.\n2004 1;79(3-4):197-223.<\/li>
  13. Wagner P, R\u00f6schke J, Mann K, Hiller W, Frank C.\nHuman sleep under the influence of pulsed radiofrequency electromagnetic\nfields: a polysomnographic study using standardized conditions.\nBioelectromagnetics: Journal of the Bioelectromagnetics Society, The Society\nfor Physical Regulation in Biology and Medicine, The European\nBioelectromagnetics Association. 1998;19(3):199-202.<\/li>
  14. Lemola S, Perkinson-Gloor N, Brand S,\nDewald-Kaufmann JF, Grob A. Adolescents\u2019 electronic media use at night, sleep\ndisturbance, and depressive symptoms in the smartphone age. Journal of youth\nand adolescence. 2015 ;44(2):405-18.<\/li>
  15. Oto R, Akda\u01e7 Z, Da\u015fda\u01e7 S, Celik Y. Evaluation\nof psychologic parameters in people occupationally exposed to radiofrequencies\nand microwave. Biotechnology & Biotechnological Equipment. 1994 J\n1;8(4):71-4.<\/li>
  16. Eger H, Jahn M. Specific symptoms and radiation\nfrom mobile basis stations in Selbitz, Bavaria, Germany: evidence for a\ndose-effect relationship (original article in German). Umwelt Med. Ges. 2010; 23:130-9.<\/li>
  17. Abdel-Rassoul G, Abou El-Fateh O, Abou Salem M,\nMichael A, Farahat F, El-Batanouny M, Salem E. Neurobehavioral effects among\ninhabitants around mobile phone base stations. Neurotoxicology. 2007\n1;28(2):434-40.<\/li>
  18. Conrad\nRH. Smart Meter Health Effects Survey and Report 2013.  https:\/\/www.mainecoalitiontostopsmartmeters.org\/wp-content\/uploads\/2013\/01\/Exhibit-10-Smart-Meter-Health-Effects-Report-Survey2.pdf\n<\/li>
  19. Kolodynski AA, Kolodynska VV. Motor and\npsychological functions of school children living in the area of the Skrunda\nRadio Location Station in Latvia. Science of the Total Environment. 1996\n2;180(1):87-93.<\/li>
  20. Shahjehan A, Afridi SA, Haider M, Iqbal A, Aziz\nS, Mansehra P. Smartphone addiction and behavioral outcomes in South Korea: A\nsystematic review and meta-analysis. Humanities & Social Sciences Reviews. 2021; 9 (2): 358-369<\/em><\/li>
  21. Siddiqi N, Jahan F, Moin F, Al-Shehhi F,\nAl-Balushi F. Excessive use of mobile phones by medical students: should\nprecautions be taken? Biomedical and Pharmacology Journal. 2017\n21;10(4):1631-8.<\/li>
  22. Lin MT, Beal MF. Mitochondrial dysfunction and\noxidative stress in neurodegenerative diseases. Nature. 2006\n19;443(7113):787-95.<\/li>
  23. Sch\u00fcz J, Waldemar G, Olsen JH, Johansen C.\nRisks for central nervous system diseases among mobile phone subscribers: a\nDanish retrospective cohort study. PLoS One. 2009 5;4(2): e4389.<\/li>
  24. Chinnery PF, Taylor GA, Howell N, Brown DT,\nParsons TJ, Turnbull DM. Point mutations of the mtDNA control region in normal\nand neurodegenerative human brains. The American Journal of Human Genetics.\n2001 1;68(2):529-32.<\/li>
  25. Gao J, Wang L, Liu J, Xie F, Su B, Wang X.\nAbnormalities of mitochondrial dynamics in neurodegenerative diseases.\nAntioxidants. 2017  5;6(2):25.<\/li>
  26. Nunomura A, Perry G, Aliev G, Hirai K, Takeda\nA, Balraj EK, Jones PK, Ghanbari H, Wataya T, Shimohama S, Chiba S. Oxidative\ndamage is the earliest event in Alzheimer disease. Journal of Neuropathology\n& Experimental Neurology. 2001 \n1;60(8):759-67.<\/li>
  27. Bender A, Krishnan KJ, Morris CM, Taylor GA,\nReeve AK, Perry RH, Jaros E, Hersheson JS, Betts J, Klopstock T, Taylor RW.\nHigh levels of mitochondrial DNA deletions in substantia nigra neurons in aging\nand Parkinson disease. Nature genetics. 2006 \n1;38(5):515-7.<\/li>
  28. Alqahtani T, Deore SL, Kide AA, Shende BA,\nSharma R, Chakole RD, Nemade LS, Kale NK, Borah S, Deokar SS, Behera A.\nMitochondrial dysfunction and oxidative stress in Alzheimer\u2019s disease, and\nParkinson\u2019s disease, Huntington\u2019s disease and amyotrophic lateral sclerosis-an\nupdated review. Mitochondrion. 2023 \n1;71:83-92.<\/li>
  29. Valenti D, de Bari L, De Filippis B, Henrion-Caude\nA, Vacca RA. Mitochondrial dysfunction as a central actor in intellectual\ndisability-related diseases: an overview of Down syndrome, autism, Fragile X\nand Rett syndrome. Neuroscience & Biobehavioral Reviews. 2014 1;46:202-17.<\/li>
  30. Helguera P, Seiglie J, Rodriguez J, Hanna M,\nHelguera G, Busciglio J. Adaptive downregulation of mitochondrial function in\ndown syndrome. Cell metabolism. 2013 \n8;17(1):132-40.<\/li>
  31. Siddiqui A, Rivera-S\u00e1nchez S, Castro MD,\nAcevedo-Torres K, Rane A, Torres-Ramos CA, Nicholls DG, Andersen JK,\nAyala-Torres S. Mitochondrial DNA damage is associated with reduced\nmitochondrial bioenergetics in Huntington’s disease. Free Radical Biology and\nMedicine. 2012  1;53(7):1478-88.<\/li>
  32. Marobbio CM, Pisano I, Porcelli V, Lasorsa FM,\nPalmieri L. Rapamycin reduces oxidative stress in frataxin-deficient yeast\ncells. Mitochondrion. 2012 \n1;12(1):156-61.<\/li>
  33. Xia H, Cao Y, Dai X, Marelja Z, Zhou D, Mo R,\nAl-Mahdawi S, Pook MA, Leimk\u00fchler S, Rouault TA, Li K. Novel frataxin isoforms\nmay contribute to the pathological mechanism of Friedreich ataxia. PLoS One. 2012;7(10):e47847.<\/li>
  34. Calabrese V, Lodi R, Tonon C, D’Agata V,\nSapienza M, Scapagnini G, Mangiameli A, Pennisi G, Stella AG, Butterfield DA.\nOxidative stress, mitochondrial dysfunction and cellular stress response in\nFriedreich’s ataxia. Journal of the neurological sciences. 2005\n15;233(1-2):145-62.<\/li>
  35. Siddiqi N, Norrish M, Heming T. Growth\nRetardation of Chick Embryo Exposed To A Low Dose Of Electromagnetic Waves.\nJournal of Ayub Medical College Abbottabad. 2016  1;28(2):224-8.<\/li>
  36. Islam MS, Islam MM, Rahman MM, Islam K. 4G\nmobile phone radiation alters some immunogenic and vascular gene expressions,\nand gross and microscopic and biochemical parameters in the chick embryo model.\nVeterinary Medicine and Science. 2023 ;9(6):2648-59.<\/li>
  37. Hasan I, Jahan MR, Islam MN, Islam MR. Effect\nof 2400 MHz mobile phone radiation exposure on the behavior and hippocampus\nmorphology in Swiss mouse model. Saudi Journal of Biological Sciences.\n2022  1;29(1):102-10.<\/li>
  38. Dasdag S\u00dc, Bilgin HM, Akdag MZ, Celik H, Aksen\nF. Effect of long-term mobile phone exposure on oxidative-antioxidative\nprocesses and nitric oxide in rats. Biotechnology & Biotechnological\nEquipment. 2008  1;22(4):992-7.<\/li>
  39. Kesari KK, Behari J, Kumar S. Mutagenic\nresponse of 2.45 GHz radiation exposure on rat brain. International Journal of\nRadiation Biology. 2010  1;86(4):334-43.<\/li>
  40. Misa-Agusti\u00f1o MJ, Leiro-Vidal JM, Gomez-Amoza\nJL, Jorge-Mora MT, Jorge-Barreiro FJ, Salas-S\u00e1nchez AA, Ares-Pena FJ,\nL\u00f3pez-Mart\u00edn E. EMF radiation at 2450 MHz triggers changes in the morphology\nand expression of heat shock proteins and glucocorticoid receptors in rat\nthymus. Life sciences. 2015  15;127:1-1.<\/li>
  41. Ozguner F, Altinbas A, Ozaydin M, Dogan A,\nVural H, Kisioglu AN, Cesur G, Yildirim NG. Mobile phone-induced myocardial\noxidative stress: protection by a novel antioxidant agent caffeic acid\nphenethyl ester. Toxicology and industrial health. 2005 ;21(7-8):223-30.<\/li>
  42. Eser O, Songur A, Akta\u015f C, Karavelio\u011flu E,\n\u00c7a\u011flar V, Aylak F, Kanter M. The effect of electromagnetic radiation on the rat\nbrain: an experimental study. Turkish neurosurgery. 2013.<\/li>
  43. Bas O, S\u00d6NMEZ O, Aslan A, Ikinci A, Hanci H,\nYildirim M, Kaya H, Ak\u00e7a M, Odaci E. Pyramidal cell loss in the cornu ammonis\nof 32-day-old female rats following exposure to a 900 Megahertz electromagnetic\nfield during prenatal days 13-21. NeuroQuantology. 2013;11(4).<\/li>
  44. Odaci E, Bas O, Kaplan S. Effects of prenatal\nexposure to a 900 MHz electromagnetic field on the dentate gyrus of rats: a\nstereological and histopathological study. Brain research. 2008  31;1238:224-9.<\/li>
  45. \u015eahin A, Aslan A, Ba\u015f O, \u0130kinci A, \u00d6zy\u0131lmaz C,\nS\u00f6nmez OF, \u00c7olako\u011flu S, Odac\u0131 E. Deleterious impacts of a 900-MHz\nelectromagnetic field on hippocampal pyramidal neurons of 8-week-old Sprague\nDawley male rats. Brain Research. 2015 \n22; 1624:232-8.<\/li>
  46. Romeo S, Zeni O, Scarf\u00ec MR, Poeta L, Lioi MB,\nSannino A. Radiofrequency electromagnetic field exposure and apoptosis: A\nscoping review of in vitro studies on mammalian cells. International Journal of\nMolecular Sciences. 2022  19;23(4):2322.<\/li>
  47. Ghazizadeh V, Naz\u0131ro\u011flu M. Electromagnetic\nradiation (Wi-Fi) and epilepsy induce calcium entry and apoptosis through\nactivation of TRPV1 channel in hippocampus and dorsal root ganglion of rats.\nMetabolic brain disease. 2014 ; 29:787-99.<\/li>
  48. Gupta SK, Mesharam MK, Krishnamurthy S.\nElectromagnetic radiation 2450 MHz exposure causes cognition deficit with\nmitochondrial dysfunction and activation of intrinsic pathway of apoptosis in\nrats. Journal of biosciences. 2018 ; 43:263-76.<\/li>
  49. Persson BR, Salford LG, Brun A. Blood\u2010brain\nbarrier permeability in rats exposed to electromagnetic fields used in wireless\ncommunication. Wireless Networks. 1997 ; 3:455-61.<\/li>
  50. Salford LG, Brun AE, Eberhardt JL, Malmgren L,\nPersson BR. Nerve cell damage in mammalian brain after exposure to microwaves\nfrom GSM mobile phones. Environmental health perspectives. 2003 ;111(7):881-3.<\/li>
  51. Stam R. Electromagnetic fields and the\nblood\u2013brain barrier. Brain research reviews. 2010 5;65(1):80-97.<\/li>
  52. Leszczynski D, Joenv\u00e4\u00e4r\u00e4 S, Reivinen J, Kuokka\nR. Non-thermal activation of the hsp27\/p38MAPK stress pathway by mobile phone\nradiation in human endothelial cells: molecular mechanism for cancer-and\nblood-brain barrier-related effects. Differentiation. 2002  1;70(2-3):120-9.<\/li>
  53. Nittby H, Brun A, Eberhardt J, Malmgren L,\nPersson BR, Salford LG. Increased blood\u2013brain barrier permeability in mammalian\nbrain 7 days after exposure to the radiation from a GSM-900 mobile phone.\nPathophysiology. 2009  1;16(2-3):103-12.<\/li>
  54. Eberhardt JL, Persson BR, Brun AE, Salford LG,\nMalmgren LO. Blood-brain barrier permeability and nerve cell damage in rat\nbrain 14 and 28 days after exposure to microwaves from GSM mobile phones.\nElectromagnetic biology and medicine. 2008 \n1;27(3):215-29.<\/li>
  55. Shabani F , Jadidi M, Esmaili M H , Sameni H R, Nazari\nH . Detrimental Effect of Mobile Phone Electromagnetic Field on Permeability of\nBlood-Brain Barrier. Middle East J Rehabil Health Stud.\n2020;7(3):e103714. <\/li>
  56. Gao P, Chen Q, Hu J, Lin Y, Lin J, Guo Q, Yue H, Zhou\nY, Zeng L, Li J, Li J, et al: Effect of ultra\u2011wide\u2011band electromagnetic pulses\non blood\u2011brain barrier permeability in rats. Mol Med Rep 2020; 22: 2775-2782.<\/li>
  57. Cho DH, Nakamura T, Lipton SA. Mitochondrial\ndynamics in cell death and neurodegeneration. Cellular and Molecular Life\nSciences. 2010 ; 67:3435-47.<\/li>
  58. Guo C, Sun L, Chen X, Zhang D. Oxidative\nstress, mitochondrial damage and neurodegenerative diseases. Neural\nregeneration research. 2013 \n25;8(21):2003-14.<\/li>
  59. Sasaki S, Warita H, Murakami T, Shibata N,\nKomori T, Abe K, Kobayashi M, Iwata M. Ultrastructural study of aggregates in\nthe spinal cord of transgenic mice with a G93A mutant SOD1 gene. Acta\nneuropathologica. 2005 ; 109:247-55.<\/li>
  60. Lahijani MS, Tehrani DM, Sabouri E.\nHistopathological and ultrastructural studies on the effects of electromagnetic\nfields on the liver of preincubated white Leghorn chicken embryo.\nElectromagnetic biology and medicine. 2009 \n1;28(4):391-413.<\/li>
  61. Siddiqi N, Al Nazwani N. Role of\nradio-frequency electromagnetic waves in causing oxidative stress. In Fundamental\nPrinciples of Oxidative Stress in Metabolism and Reproduction 2024  1 (pp. 77-91). Academic Press.<\/li>
  62. Douarre C, Sourbier C, Dalla Rosa I, Brata Das\nB, Redon CE, Zhang H, Neckers L, Pommier Y. Mitochondrial topoisomerase I is\ncritical for mitochondrial integrity and cellular energy metabolism. PloS one.\n2012  20;7(7): e41094.<\/li>
  63. Sas K, Robotka H, Toldi J, V\u00e9csei L.\nMitochondria, metabolic disturbances, oxidative stress and the kynurenine\nsystem, with focus on neurodegenerative disorders. Journal of the neurological\nsciences. 2007  15;257(1-2):221-39.<\/li>
  64. Chen JQ, Yager JD, Russo J. Regulation of\nmitochondrial respiratory chain structure and function by estrogens\/estrogen\nreceptors and potential physiological\/pathophysiological implications.\nBiochimica et Biophysica Acta (BBA)-Molecular Cell Research. 2005  30;1746(1):1-7.<\/li>
  65. Wei YH, Lu CY, Wei CY, Ma YS, Lee HC. Oxidative\nstress in human aging and mitochondrial disease-consequences of defective\nmitochondrial respiration and impaired antioxidant enzyme system. Chinese\nJournal of Physiology. 2001 \n31;44(1):1-2.<\/li>
  66. Finkel T, Holbrook NJ. Oxidants, oxidative\nstress and the biology of ageing. nature. 2000 \n9;408(6809):239-47.<\/li>
  67. Andreazza AC, Shao L, Wang JF, Young LT.\nMitochondrial complex I activity and oxidative damage to mitochondrial proteins\nin the prefrontal cortex of patients with bipolar disorder. Archives of general\npsychiatry. 2010  1;67(4):360-8.<\/li>
  68. Van Houten B, Woshner V, Santos JH. Role of\nmitochondrial DNA in toxic responses to oxidative stress. DNA repair. 2006  3;5(2):145-52.<\/li>
  69. Joshi G, Sultana R, Perluigi M, Butterfield DA.\nIn vivo protection of synaptosomes from oxidative stress mediated by Fe2+\/H2O2\nor 2, 2-azobis-(2-amidinopropane) dihydrochloride by the glutathione mimetic\ntricyclodecan-9-yl-xanthogenate. Free Radical Biology and Medicine. 2005  15;38(8):1023-31.<\/li>
  70. Ghezzi D, Zeviani M. Assembly factors of human\nmitochondrial respiratory chain complexes: physiology and pathophysiology.\nMitochondrial Oxidative Phosphorylation: Nuclear-Encoded Genes, Enzyme\nRegulation, and Pathophysiology. 2012 \n11:65-106.<\/li>
  71. Ross WN. Understanding calcium waves and sparks\nin central neurons. Nature Reviews Neuroscience. 2012 ;13(3):157-68.<\/li>
  72. Beason RC, Semm P. Responses of neurons to an\namplitude modulated microwave stimulus. Neuroscience Letters. 2002  29;333(3):175-8.<\/li>
  73. Krey JF, Dolmetsch RE. Molecular mechanisms of\nautism: a possible role for Ca2+ signaling. Current opinion in neurobiology.\n2007  1;17(1):112-9.<\/li>
  74. Volkow ND, Wang GJ, Fowler JS, Tomasi D, Telang\nF. Addiction: beyond dopamine reward circuitry. Proceedings of the National\nAcademy of Sciences. 2011 \n13;108(37):15037-42.<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

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