The Radiofrequency Radiation Effect of Mobile Phone on the neurons and glial cells of the murine hippocampus

Dhiraj Maskey,Myeung Ju Kim 대한체질인류학회 2011 대한체질인류학회 학술대회 연제 초록 Vol.54 No.-

Alteration of glycine receptor immunoreactivity in the auditory brainstem of mice following three months of exposure to radiofrequency radiation at SAR 4.0 W/kg

MASKEY, DHIRAJ,KIM, HYUNG GUN,SUH, MYUNG-WHAN,ROH, GU SEOB,KIM, MYEUNG JU D.A. Spandidos 2014 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.34 No.2

<P>The increasing use of mobile communication has triggered an interest in its possible effects on the regulation of neurotransmitter signals. Due to the close proximity of mobile phones to hearing-related brain regions during usage, its use may lead to a decrease in the ability to segregate sounds, leading to serious auditory dysfunction caused by the prolonged exposure to radiofrequency (RF) radiation. The interplay among auditory processing, excitation and inhibitory molecule interactions plays a major role in auditory function. In particular, inhibitory molecules, such a glycine, are predominantly localized in the auditory brainstem. However, the effects of exposure to RF radiation on auditory function have not been reported to date. Thus, the aim of the present study was to investigate the effects of exposure to RF radiation on glycine receptor (GlyR) immunoreactivity (IR) in the auditory brainstem region at 835 MHz with a specific absorption rate of 4.0 W/kg for three months using free-floating immunohistochemistry. Compared with the sham control (SC) group, a significant loss of staining intensity of neuropils and cells in the different subdivisions of the auditory brainstem regions was observed in the mice exposed to RF radiation (E4 group). A decrease in the number of GlyR immunoreactive cells was also noted in the cochlear nuclear complex [anteroventral cochlear nucleus (AVCN), 31.09%; dorsal cochlear nucleus (DCN), 14.08%; posteroventral cochlear nucleus (PVCN), 32.79%] and the superior olivary complex (SOC) [lateral superior olivary nucleus (LSO), 36.85%; superior paraolivary nucleus (SPN), 24.33%, medial superior olivary nucleus (MSO), 23.23%; medial nucleus of the trapezoid body (MNTB), 10.15%] of the mice in the E4 group. Auditory brainstem response (ABR) analysis also revealed a significant threshold elevation of in the exposed (E4) group, which may be associated with auditory dysfunction. The present study suggests that the auditory brainstem region is susceptible to chronic exposure to RF radiation, which may affect the function of the central auditory system.</P>

The Effect of 835 ㎒ Radiofrequency Radiation Exposure on the Immunohistochemical Distribution of Calbindin D28k and Calretinin in the Mouse Cerebellar Cortex

Dhiraj Maskey(마스키 디라즈),In-young Choi(최인영),Ki-Sup Park(박기섭),Jonu Pradhan(프라단 조누),Young Bok Yoo(유영복) 대한체질인류학회 2009 해부·생물인류학 (Anat Biol Anthropol) Vol.22 No.4

현대 널리 보급된 무선 휴대폰의 사용은 이로 인해 발생하는 전자기파가 뇌와 신경계통에 미칠 생물학적 영향은 중요 관심사이지만, 아직까지 많은 연구가 진행되지 않았다. 전자기파의 방출로 인해 신경계통에 미치는 영향으로 여러 연구에서 뇌의 여러 부분에서의 신경세포수의 감소, 신경세포의 손상 등을 보고하였다. 그러나, 아직까지 행동을 관장하는 소뇌부분에 관한 어떤 연구도 진행되지 않아 본 연구에서 생체 항상성을 유지하는 데 중요한 이온인 칼슘의 물질대사에 중요한 역할을 하는 calbindin D28k (CB)와 calretinin (CR)의 항체를 이용하여 면역 염색학적인 방법으로 생쥐의 소뇌에서 835 ㎒전자기파 방출이 미치는 영향을 조사하였다. CB와 CR 특이 항체에 대한 면역염색성은 835 ㎒의 전자기장에 서로 다른 시간 동안 노출시킨 후 시행하였는데, 사용한 실험조건은 1.6W/㎏ 에서 하루 1시간씩 5일 동안 노출된 군, 4.0W/㎏에서 하루 1시간씩 5일 동안 노출된 군, 1.6W/㎏에서 하루 5시간씩 하루 동안 노출된 군, 4.0W/㎏에서 하루 5시간씩 하루 동안 노출된 군, 그리고 1.6 W/㎏ 에서 한달 동안 8시간씩 매일 노출된 군을 이용했다. CB의 면역반응성은 조롱박세포에서 현저하게 관찰되었으며, 가지돌기와 먼쪽 축삭들에서도 관찰되었다. 1.6W/㎏에서 5일 동안 전자기파에 노출된 경우 CB의 면역반응성이 감소하였고, CR의 면역반응성은 주로 과립층의 작은세포들에 국한되었으며, 면역반응성은 한 달간 노출된 군에서 염색성이 증가되었으며, 조롱박세포층에서는 어떤 세포도 염색된 것을 관찰할 수 없었다. 따라서, 본 연구는 835 ㎒ 전자기파에 장기간 지속적으로 생체가 노출되면 신경계통에서 물질대사의 항상성을 유지하는 칼슘농도의 변화를 초래해, 소뇌에서의 칼슘대사 이상으로 인한 병적인 행동을 유발할 수 있는 가능성을 제시하였다. Widespread use of mobile phones and subsequent electromagnetic field (EMF) exposure have raised crucial question of their possible biological effects on the nervous system. The study on the effect of radiofrequency (RF) radiation on the nervous system, however, did not precede enough to determine the biological hazard to brain. Until now, several studies have reported decreases in neuron number and neuronal damage in the cortex, hippocampus, and basal ganglia in the brains of animals exposed to RF radiation. However, there were few reports about the cerebellum, the main voluntary motor control center. In this regard, by using immunohistochemisty, current study intended to investigate the changes in the calbindin D28k (CB) and calretinin (CR)-immunoreactivity (IR) in the mouse cerebellar cortex after EMF exposure at 835 ㎒ for different exposure times and absorption rates, 1h/day for 5 days at 1.6 W /㎏, 1 h/day for 5 days at 4.0 W /㎏, 5 h/day for 1 day at 1.6 W /㎏, 5 h/day for 1 day at 4.0 W /㎏, daily exposure for one month at 1.6 W /㎏. Among groups, most prominent CB IR was observed in the Purkinje cell layer followed by molecular and granular layer. The highest CB IR was noted in 5 h/day for 1 day at 1.6 W /㎏ in the entire three layers while the lowest was noted in one month at 1.6 W /㎏. Similarly CR IR was maximum in one month at 1.6 W /㎏ whilst the lowest was observed in 1 h/day for 5 days at 4.0W/㎏. EMF exposure for 5 days at 1.6 W/㎏ reduced CB-IR. The CR-IR was mainly localized in small cells in the granular layer, with maximum IR observed after one month exposure. Therefore, the present study suggest the possibility of alterations of calcium ion concentration, which playa role in maintaining metabolic homeostasis, in the cerebellum after long-term exposure to 835 ㎒ of RF radiation, which might lead to the disruption of normal trait.

Effect of ginseng extracts on 835 ㎒ radiofrequency radiation exposure on calcium binding proteins in the mouse brain

Dhiraj Maskey,Bijay Aryal,Jae Won Yang,Myeung Ju Kim,Hyung Gun Kim(김형건) 고려인삼학회 2009 고려인삼학회 학술대회 Vol.2009 No.12

Preventive Effect of Ginseng Saponin on Hearing Loss Against Short Term Radiofrequency Exposure in the Superior Olivary Complex of Mice

Dhiraj Maskey,Hyung-Gun Kim 고려인삼학회 2011 고려인삼학회 학술대회 Vol.2011 No.4

한국인 태아 두피 털주머니 발생과정에서 bcl-2의 발현

마스키 디라즈(Dhiraj Maskey),최인영(In-young Choi),박기섭(Ki-Sup Park),프라단 조누(Jonu Pradhan),유영복(YoungBok Yoo) 대한해부학회 2009 Anatomy & Cell Biology Vol.42 No.4

발생 14-24주의 한국인 태아 두피조직을 대상으로 아포프토시스를 억제하는 유전자인 bcl-2가 털주머니(hair follicle) 발생에 미치는 영향을 파악하고자 고집적 (high-throughput) 연구방법인 tissue-array 기법으로 면역조직화학염색을 이용하여 관찰하였다, 발생 14주에서 24주까지 표피의 바닥세포에서 bcl-2가 발현되었고 bcl-2가 발현된 멜라닌세포는 표피의 바닥층, 바닥위층, 털바탕질세포 사이와 바깥상피뿌리집에서 관찰되었다. 발생 초기나 세포가 계속 생성되는 부위에서 미성숙한 세포가 bcl-2를 발현하여 세포 사망이 억제되나 분화가 진행되면서 bcl-2 발현의 감소로 세포 사망이 증가하여 조직의 형태형성이나 기능 완성에 기여하는 것이 사람의 태아 발생기 털주머니에서도 확인할 수 있었다. This study was intended to understand the role of the apoptosis-suppressing gene, bcl-2, in the hair follicle development. Immunohistochemistry for bcl-2 was performed using a high-throughput tissue-array technique, on Korean fetal scalp tissues at the 14 weeks, 16 weeks, 19 weeks & 24 weeks of the development. The results showed that the basal cells of epidermis were stained from 14 weeks to 24 weeks and the immunoreactive melanocytes were observed in the basal layer and suprabasal layer of epidermis as well as in the hair matrix cells and the external root sheath. At 19 weeks, the follicles at all stages of morphogenesis were observed. In the early stages, the epithelial cells of hair germ and hair peg, the mesenchymal cells surrounding them were stained. In the more advanced stage, the bcl-2 expression of follicular epithelial cells diminished to allow keratinization, hair canal formation and holocrins secretion to take place. In the bulbous hair peg stage, the hair papilla cells, the hair matrix cells, the basal cells of the primitive sebaceous gland, the primitive arrector pilli and the basal cells of the external root sheath were stained. We confirmed in the developing hair follicle that in the early stage or in the place where the cells continued to proliferate, the immature cells expressed bcl-2 to suppress cell death to overcome the susceptibility to cell death and when the differentiation was being achieved, the reduction of bcl-2 expression increased cell death to perform the tissue morphogenesis or the organ functions.

Decreased Immunoreactivities and Functions of the Chloride Transporters, KCC2 and NKCC1, in the Lateral Superior Olive Neurons of Circling Mice

Jonu Pradhan,Dhiraj Maskey,박기섭,김명주,안승철 대한이비인후과학회 2011 Clinical and Experimental Otorhinolaryngology Vol.4 No.1

Objectives. We tested the possibility of differential expression and function of the potassium-chloride (KCC2) and sodiumpotassium-2 chloride (NKCC1) co-transporters in the lateral superior olive (LSO) of heterozygous (+/cir) or homozygous (cir/cir) mice. Methods. Mice pups aged from postnatal (P) day 9 to 16 were used. Tails from mice were cut for DNA typing. For Immunohistochemical analysis, rabbit polyclonal anti-KCC2 or rabbit polyclonal anti-NKCC1 was used and the density of immunolabelings was evaluated using the NIH image program. For functional analysis, whole cell voltage clamp technique was used in brain stem slices and the changes of reversal potentials were evaluated at various membrane potentials. Results. Immunohistochemical analysis revealed both KCC2 and NKCC1 immunoreactivities were more prominent in heterozygous (+/cir) than homozygous (cir/cir) mice on P day 16. In P9–P12 heterozygous (+/cir) mice, the reversal potential (Egly) of glycine-induced currents was shifted to a more negative potential by 50 μM bumetanide, a known NKCC1 blocker, and the negatively shifted Egly was restored by additional application of 1 mM furosemide, a KCC2blocker (-58.9±2.6 mV to -66.0±1.5 mV [bumetanide], -66.0±1.5 mV to -59.8±2.8 mV [furosemide+bumetanide],n=11). However, only bumetanide was weakly, but significantly effective (-60.1±2.9 mV to -62.7±2.6 mV [bumetanide],-62.7±2.6 mV to -62.1±2.5 mV [furosemide+bumetanide], n=7) in P9–P12 homozygous (cir/cir) mice. Conclusion. The less prominent immunoreactivities and weak or absent responses to bumetanide or furosemide suggest impaired function or delayed development of both transporters in homozygous (cir/cir) mice.

Effect of Ginseng on Calretinin Expression in Mouse Hippocampus Following Exposure to 835 MHz Radiofrequency

Aryal, Bijay,Maskey, Dhiraj,Kim, Myeung-Ju,Yang, Jae-Won,Kim, Hyung-Gun The Korean Society of Ginseng 2011 Journal of Ginseng Research Vol.35 No.2

Exponential rise in the use of mobile communication devices has generated health concerns due to radiofrequency (RF) exposure due to its close proximity to the head. Calcium binding proteins like calretinin regulate the levels of calcium ($Ca^{2+}$) which plays an important role in biological systems. Ginseng is known for maintaining equilibrium in the human body and may play a beneficial radioprotectant role against electromagnetic field (EMF) exposure. In the present study, we evaluated the radioprotective effects of red ginseng (RG) extract in a mouse model. Calretinin (CR) expression was measured using a free-floating immunohistochemical method in the hippocampus of mice after 835 MHz EMF exposure for 5 h/d for 5 d at specific absorption rate=1.6 W/kg for the different experimental groups. The control animals were treated with NaCl while the experimental animals received 10 mg/kg ginseng, or 30 mg/kg; EMF exposed mice were also treated with NaCl, 10 mg/kg ginseng (E10), or 30 mg/kg (E30). Decreases in CR immunoreactivity (IR) along with loss of CA1 and CA3 interneurons and infragranular cells were observed in the ENaCl group while such losses were not observed in the E10 and E30 groups. CR IR significantly increased in the RG-treated group compared to control and EMF-exposed groups treated with NaCl. The study demonstrates that RG extract can serve as a radioprotective agent that maintains $Ca^{2+}$ homeostasis and prevents neuronal loss in the brain hippocampal region caused by RF exposure.

Effect of Ginseng on Calretinin Expression in Mouse Hippocampus Following Exposure to 835 ㎒ Radiofrequency

Bijay Aryal,Dhiraj Maskey,Myeung-Ju Kim,Jae-Won Yang,Hyung-Gun Kim 고려인삼학회 2011 Journal of Ginseng Research Vol.35 No.2

Exponential rise in the use of mobile communication devices has generated health concerns due to radiofrequency (RF) exposure due to its close proximity to the head. Calcium binding proteins like calretinin regulate the levels of calcium (Ca²?) which plays an important role in biological systems. Ginseng is known for maintaining equilibrium in the human body and may play a beneficial radioprotectant role against electromagnetic field (EMF) exposure. In the present study, we evaluated the radioprotective effects of red ginseng (RG) extract in a mouse model. Calretinin (CR) expression was measured using a free-floating immunohistochemical method in the hippocampus of mice after 835 ㎒ EMF exposure for 5 h/d for 5 d at specific absorption rate=1.6 W/㎏ for the different experimental groups. The control animals were treated with NaCl while the experimental animals received 10 ㎎/㎏ ginseng, or 30 ㎎/㎏; EMF exposed mice were also treated with NaCl, 10 ㎎/㎏ ginseng (E10), or 30 ㎎/㎏ (E30). Decreases in CR immunoreactivity (IR) along with loss of CA1 and CA3 interneurons and infragranular cells were observed in the ENaCl group while such losses were not observed in the E10 and E30 groups. CR IR significantly increased in the RG-treated group compared to control and EMF-exposed groups treated with NaCl. The study demonstrates that RG extract can serve as a radioprotective agent that maintains Ca²? homeostasis and prevents neuronal loss in the brain hippocampal region caused by RF exposure.

Effect of Ginseng on Calretinin Expression in Mouse Hippocampus Following Exposure to 835 MHz Radiofrequency

Bijay Aryal,Dhiraj Maskey,김명주,양재원,김형건 고려인삼학회 2011 Journal of Ginseng Research Vol.35 No.2

Exponential rise in the use of mobile communication devices has generated health concerns due to radiofrequency (RF) exposure due to its close proximity to the head. Calcium binding proteins like calretinin regulate the levels of calcium (Ca^(2+)) which plays an important role in biological systems. Ginseng is known for maintaining equilibrium in the human body and may play a beneficial radioprotectant role against electromagnetic field (EMF) exposure. In the present study, we evaluated the radioprotective effects of red ginseng (RG) extract in a mouse model. Calretinin (CR) expression was measured using a free-floating immunohistochemical method in the hippocampus of mice after 835 MHz EMF exposure for 5 h/d for 5 d at specific absorption rate=1.6 W/kg for the different experimental groups. The control animals were treated with NaCl while the experimental animals received 10 mg/kg ginseng, or 30 mg/kg; EMF exposed mice were also treated with NaCl, 10 mg/kg ginseng (E10), or 30 mg/kg (E30). Decreases in CR immunoreactivity (IR) along with loss of CA1 and CA3 interneurons and infragranular cells were observed in the ENaCl group while such losses were not observed in the E10 and E30 groups. CR IR significantly increased in the RG-treated group compared to control and EMF-exposed groups treated with NaCl. The study demonstrates that RG extract can serve as a radioprotective agent that maintains Ca^(2+) homeostasis and prevents neuronal loss in the brain hippocampal region caused by RF exposure.