Subchronic Oral Dose Toxicity Study of Enterococcus Faecalis 2001 (EF 2001) in Mice

Yeun-Hwa Gu,Takenori Yamasita,Ki-Mun Kang 한국독성학회 2018 Toxicological Research Vol.34 No.1

As a part of general toxicity studies of Enterococcus Faecalis 2001 (EF 2001) prepared using heat-treatment bacillus mort body EF 2001 in mice, this study examined the toxicity of EF 2001 in single and repeated administrations following the previous report in order to apply this product to preventive medicine. The safety of oral ingestion of EF 2001 was examined in 6-week-old male and female ICR mice with 1,000 mg/kg, 3,000 mg/kg and 5,000 mg/kg body weight/day administrated by gavage of the maximum acceptable dose of EF 2001. The study was conducted using distilled water as a control following the methods for general toxicity studies described in the “Guidelines for Non-clinical Studies of Pharmaceutical Products 2002”. As a control, 1) observation of general conditions, 2) measurement of body weight, 3) determination of food consumption, 4) determination of water consumption, 5) blood test and urinalysis and 6) pathological examination were performed for the administration of EF 2001. Mice received EF 2001 for 13 weeks and results were compared with those of the control group that received distilled water. The results of the above examinations revealed no significant differences between control and EF 2001 groups for both males and females. Thus, no notable toxicity was confirmed with single and repeated oral administrations of EF 2001. Oral administration in the above doses did not result in abnormal symptoms or death during the observation period. No abnormalities in blood cell count or organ weights were seen. Without any evidence of toxicity to cells and organs, EF 2001 is speculated to not adversely affect living organisms. The 50% lethal dose of EF 2001 with oral administration in mice is estimated to be greater than 5,000 mg/kg body weight/day for both male and female mice. Therefore, LD50 value for animals was 5,000 mg/kg or more.

Effect of Lepidium Meyenii Walp and Jatropha Macrantha for Estradiol-17β, Progesterone and Testosterone in Serum Concentration, and For Conception Rate and Number of Fetuses in Mice

Gu, Yeun-Hwa,Oshima, Masami,Tsukada, Sekihito,Suzuki, Ikukatsu,Baba, Hiroyuki,Hasegawa, Takeo INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2002 JOURNAL OF ORIENTAL MEDICINE Vol.7 No.1

To determine the effect of Pervian herb medicines which were called Lepidium meyenii Walp and Jatropha macrantha for blood concentration Estradiol-17β, Progesterone and Testosterone and for conception rate and number of fetuses in mice. Forty mice were keeping ten pieces of each cages and four groups and so Lepidium meyenii Walp and Jatropha macrantha were giving instead of water to mice for thirty days. The blood samples were collected at the final day and these samples were measured Estradiol-17β, Progesterone and Testosterone by using Radio Immuno Assay (RIA). The conception rate and number of fetuses studies were mating at the final day for three hours. Then female mice were confirmed the vaginal plug and number of fetuses fifteen days after. Results for Estradiol-17β concentration, J group was shown higher than C group. As for L group, Progesterone concentration showed clearly higher than C group with in comparison (p<0.05). For Testosterone concentration, L group and LJ group were shown clearly higher than C group (p<0.05). Results for conception rate and number of fetuses were as same as Progesterone concentration results. Lepidium meyenii Walp and Jatropha macrantha had the effect to increase for Estradiol-17β, Progesterone and Testosterone of blood concentration and for conception rate and number of fetuses.

A Study of the radioprotection effect of macro-glucan (β-1-3glucan) on thefetuses of ICR mice

Gu, Yeun-hwa,Hasegawa, Takeo,Suzuki, Ikukatsu,Hayashi, Ikuo,Ahn, Kyoo-Seok,Tawaraya, Hitoshi INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2000 JOURNAL OF ORIENTAL MEDICINE Vol.5 No.1

In this study abundant malformations of ICR mice in the organogenesis stages were observed and the radioprotection effect of Macro-Glucan(β-1-3glucan) was shown. This research examined the radioprotection effect of Macro-Glucan(β-1-3glucan) and its teratogenic properties. The reason it has a preventive effect on a tall fetus was examined in the research. We examined the determination stage (organogenesis), and pointed to malformation toward the radiation in the enviornmental agent to the malformaiton. Moreover, we examined the cellularis level of the embryos toward the radiation and analyzed the mechanism of the protection effect of the Macro-Glucan(-1-3glucan) from malformation induction by the radiation. Therefore, the purpose of this research is to get a foundation of data of the radioprotection effect of Macro-Glucan(-1-3glucan). This study produced the following results : It was made clear that Macro-Glucan(-1-3glucan) had a radioprotection effect against embryonic death. Radioprotection effect was recognized as the teratogenesis rate by Macro-Glucan(-1-3glucan)'s administering it before the exposure. Therefore, the subject faced radiation exposure by the Macro-Glucan(-1-3glucan) administration, and radiological protecting effect was made clear.

Radioprotection Effects to the Radiation Side Effect in EEM (Extracts of Edible Mushrooms)

Gu, Yeun-Hwa,Masaki, Matsumori,Suzuki, Ikukatu,Takeo, Hasegawa,Yuuichi, Ukawa,Hitoshi, Tawaraya,Ikuo, Hayashi,Ahn, Kyoo-Seok INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2003 JOURNAL OF ORIENTAL MEDICINE Vol.6 No.1

Presently, raditherapy is often used alone or in combination with anti-cancer medicine as a treatment for cancer. Chemosynthesis has been used as a radiation protection medicine. But, its use has been restricted due to serious side effects. ICR mice have often been used for malformation experiments as they were used in this research. Here, EEM (extracts of edible mushrooms (and extraction from Flammulina velutires and lyuophyllum ulmarium)), a natural product, was used as a radioprotective agent and its effects were examined. A decline in the fetus malformation rate was recognized when giveing EEM before the radiation irradiation. Moreover, a repression in the decrease of fetus weight was recognized, and the effect on radiation protection of EEM was explained at the individual level. And, I had the effect on protection that faced a corpuscle cell by the radiation by giving EEM was explained in the same way too. As for the cell level as well, it was proved that there was an effect on protection against radiation irradiation in the same way as the individual level. Furthermore, an examination was done about the effects on the external malformation to the radiation, the embryonic death rate and the sensitivity of the lymphocyte at the organogenesis stage. Furtheromore, the study was done about the effects on the malformation to the radiation, the embryonic death rate and the sensitivity of the lymphocyte at the fetus of the organogenesis stage.

The Effect of Rice Roast Against the Immunity Precipitation in the Radiation

Gu, Yeun-Hwa,Suzuki, Ikukatsu,Park, Sang-Rae,Hasegawa, Takeo,Tatehana, Tieko INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2001 JOURNAL OF ORIENTAL MEDICINE Vol.6 No.1

Recently, it was cleared that the injury that it oxidized by super oxygen, free radical in the various diseases and aging was deeply concerned. Moreover, it pays attention to the prevention by the anti-oxide, and research on the anti-oxide quality is composed of is formed in the living body, and it oxidizes in lipids, protein, nuclear acid radiation irradiation damage. Therefore, it knows that various organization difficulties are pulled up. Actually, there is a report of controlling the lipids of over-oxidization when an X-rays and a gamma ray were irradiated, and DNA damage by the treatment of the vitamin E. It considers that RICE ROAST plays a part as radical scavenger that super oxygen that occurred in the body is erased, too, from the result of this research as well. And, there is MEA (cysteamine hydrochloride) as one of the radiation protection medicine. Watanabes report that the DNA one chain cutting caused by a gamma ray, and chromosome aberration were decreased remarkably after the radiation irradiation as a result of examining the effect of the vitamin E by using the embryonic cells of the ham star against the cell damage. This effect is thought to be effect on prevention of OH radical. Therefore, radical scavenger prevention action due to the radiation chemical action and effect on anti-oxidization were explained in this research in the same way as the number of leucocyte. It faced the number of the occurrences of lymphocyte, effect on radiation protection by the RICE ROAST medication that a radiation irradiates it was explained. The result of the blood cell level study were explained Therefore, it thinks about damage by the radiation by radical of super oxygen with the thing due to the repression action of the vitamin E due to the oxide target damage of nuclear acid, the lipids, the protein in the living body. The radical scavenger prevention action due to the radiation che.

Radiation Protection Effect on Propolis and Agaricus Combination

Gu, Yeun-hwa,Suzuki, Ikukatsu,Park, Sangrae,Hasegawa, Takeo,Koike, Mtsuo INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2001 JOURNAL OF ORIENTAL MEDICINE Vol.6 No.1

Damage is given to the marrow which is a place to make the cell for which to administer immunity such as a leukocyte, lymphocyte, monocyte when a X-ray is irradiated, and it knows that that cellular number decreases. It develops into leukocyte decrease syndrome, the symptoms such as immunity in severity case. This time, the effect that controlled a decrease such as a leukocyte due to the X-ray irradiation, a lymphocyte, monocyte was admitted by the medication of Propolis and agaricus. It thinks with the thing that the effect is shown for the effect on immunity recovery in the radiotherapy and the prevention of a side effect of the radiation from this result. Furthermore, increase in the effect on immunity power recovery at the time of the immunity power decline by the anti-cancer medicine medication and the effect on an anti-tumor of the anticancer medicine, prevention of a side effect of the anticancer medicine, and so on can be expected. Damage is given to the marrow which is a place to make the cell for which to administer innumity such as a leukocyte, lymphocyte, monocyte when an X-ray is irradiated, and it knows that that cellular number decreases. It develops into leukocyte decrease syndrome, the symptoms such as immunity in severity case. This time, the effect that controlled a decrease such as a leukocyte due to the X-ray irradiation, a lymphocyte, monocyte was admitted by the medication of propolis and agaricus. It thinks with the thing that the effect is shown for the effect on immunity recovery in the radiotherapy and the prevention of a side effect of the radiation from this result. Futhermore, increase in the effect on immunity power recovery at the time of the immunity power decline by the anti-tumor of the anticancer medicine, prevention of a side effect of the anticancer medicine, and so on can be expected.

The Developmental Effects of Radiation on ICR Mouse Embryos in Preimplantation Stage

Gu,Yeun Hwa 대한방사선 방어학회 1996 방사선방어학회지 Vol.21 No.4

着床前期의 胎兒는 放射線을 비롯한 많은 環境要因에 대하여 感受性이 높은 個體임에도 불구하고 특히 이 시기는 姙娠婦가 自覺적으로 임신을 감지할 수 없는 시기이기에 이러한 여러 환경유해요인으로 부터 의도적으로 피할 수가 없다. 그러므로 착상전기의 영향을 충분히 검토한 후에 의료행위를 취할 것이며 이에 대한 防禦대책도 검토할 필요가 있다. 종래 까지 방사선에 대한 태아영향에 많은 연구결과에 의하면 방사선 및 그 외의 유해요인에 대한 착상전기의 영향은 胚死亡(流産)만이 일어나며 奇形은 誘發하지 않는다고 하여 發生學등 여러 교과서에서 기형은 器官形成期만이 局限해서 일어나는 영향이라고 단정되어 왔었다. 그러나 이 연구결과 착상전기에 있어서도 기형이 유발하여 오히려 器官形成期보다도 감수성이 높다는 것이다. 또한 착상전기에서도 기형유발의 시기특이성을 가지며 여러 종류의 기형이 발생한다는 것이 본 연구로부터 밝혀졌다. 실험동물은 ICR Mouse를 사용했다. ICR Mouse는 일반적으로 태아사망 및 기형실험에 널리 사용되는 것이다. 사육조건은 Conventional 한 조건하에서 사육했으며 Mating 方法은 Female 마우스의 발정기(Sexual Excitement period)에 있는 mouse 질(膣)을 육안 적으로 관찰하여 AM. 6:00∼9:00시까지 3시간만 mate시켰다. AM 9:00시에 Vaginal Plug를 관찰하여 임신을 확인했다. Plug가 확인된 마우스는 AM 8:00시에 수정(Conception)된 것으로 가정하고 이 시점을 임신 0링 0시로 수정 난의 태아연령을 산정했다. 방사선조사는 137 Cs Υ-선을 사용하였으며 임신 마우스의 전신조사를 실시하고 조사한 시기는 각 2, 48, 72, 96hpc이며 조사한 방사선 선량 군은 0.1∼2.5Gy이다. 태아영향 관찰지표는 태아 연령은 mate일 오전 8:00시를 임신 0일 0시로 환산하여 태아연령 18일에 임신마우스를 Cervical vertebral dislocation에 의해 도살했다. 도살후 해부하여 각 임신 마우스별로 관찰했다. 착상 율을 관찰하기 위하여 황체수를 세었고, 태아사망과 생존태아를 구별했다. 자궁내 사망의 분류는 태아사망을 ① preimplantation death ② Embryonic death ③ Fetal death로 분류했다. 착상전사망은 수정후 0∼4.5日(1세포기∼배반포후기 부화까지)까지의 사망으로써 난소수 황체수(배란 수)와 착상태아(생존태아, 착상흔, 태반유잔, 흡수태아, 침연태아의 합계)로 부터 구할 수 있다. Embryonic death는 수정후 4.5日∼13日까지의 사망으로써 Implantation sites, Placental remnants, Resorption of fetus로 관찰된 것이다. Fetal death는 수정후 14日∼18日까지의 사망으로써 Maceration of fetus로 관찰되는 것이다. 통계학적 분석은 각 Group의 착상 율과 자궁내사망율을 산출할 때에는 각 임신마우스에 따라 발생빈도가 크게 다르기 때문에 통계처리에는 Non parametric 검정인 Kluskal Wallis 검정을 사용하여 분석하였다. 또한 개체 Level 영향인 착상율, 태아사망, 기형의 threshold dose의 산정에 대해서는 SAS-Logistic 검정에 따라 통계분석을 하여 5%(Ld5, ED5) 및 10%×2/3점을 threshold dose로 판단했다. 태아체중에 대해서는 parametric검정인 t-test검정에 의하여 분석했다. 그 결과 착상전기에서도 기형이 유발하며 특히 시기에 따라 일어나는 때와 일어나지 않는 때가 있음을 본 연구로부터 밝혀졌다. 또한 착상전기의 영향으로써 유발되는 기형은 여러 종류의 기형이 발생함이 밝혀졌다. 특히 이시기는 착상전사망 및 胚사망은 방사선 선량에 따라 크게 일어나나 태아사망(Fetal death) 및 태아체중은 有意差가 없었다. Embryos and fetuses are more sensitive to various environmental agents than are adults or children. The biological effects such as intrauterine death and malformation are closely connected with prenatal exposure very various agents. The sensitivity of these embryonic/fetal effects depends on the stage of pregnancy. From the viewpoint of fetal development, embryonic and fetal stages can be divided into three stages : Preimplantation, organogenetic and fetal. Each stage corresponds to 0 to 4.5days, 4.5 to 13.5days, and 13.5days of gestation in mice, respectively. Many studies on the biologcal effects of mice irradiated by γ-rays at various stages during organogenesis and fetal period have been performed. Based on these results, the dose-effect and dose-response relationships in malformations, intrauterine death, or retardation of the physical growth have been practically modeled by the ICRP(International Commission on Radiological Protection) and other international bodies for radiation protection. Many experimental studies in mice have made it clear that mice embryos in the preimplantation period have a higher sensitivity to radiation for lethal effects than the embryos/fetuses on other prenatal periods. However, no teratogenic effects of radiation at preimplantation stages of mice have been described in many textbooks. It has been believed that "all or none action results" for radiation of mice during the preimplantation period were applied. The teratogenic and lethal effects during the preimplantation stage are one of the most important problems from the viewpoint of radiological protection, since the preimplantation stage is the period when the pregnancy itself is not noticed by a pregnant woman. There are many physical or chemical agents which affect embryos/fetuses in the environment. It is assumed that each agents indirectly effects a human. Then, a safety criterion on each agent is determined independently. The pregnant ICR mice on 2, 48, 72 or 96 hours post-conception (hpc), at which are preimplantation stage of embryos, were irradiated whole body Cesium-gamma radiation at doses of 0.1, 0.25, 0.5, 1.5 and 2.5 Gy with dose rate of 0.2 Gy/min. In the embryos from the fetuses from the mice irradiated at various period in preimplantation, embryonic/fetal mortalities, incidence of external gross malformation, fetal body weight and sex ratio were observed at day 18 of gestation. The sensitivity of embryonic mortalities in the mice irradiated at the stage of preimplantation were higher than those in the mice irradiated at the stage of organogenesis. And the more sensitive periods of preimplantation stage for embryonic death were 2 and 48 hpc, at which embryos were one cell and 4 to 7 cell stage, respectively. Many types of the external gross malformations such as exencephaly, cleft palate and anophthalmia were observed in the fetuses from the mice irradiated at 2, 72 and 96 hpc. However, no malformations were observed in the mice irradiated at 48 hpc, at which stage the embryos were about 6 cell stage precompacted embryos. So far, it is believed that the embryos on preimplantation stage are not susceptible to teratogens such as radiation and chemical agents. In this study, the sensitivity for external malformations in the fetuses from the mice irradiated at preimplantation were higher than those in the fetuses on stage of organogenesis.

The Development Effects of Radiation on ICR Mouse Embryos in Preimplantation Stage

Yeun Hwa Gu 대한방사선방어학회 1996 방사선방어학회지 Vol.21 No.4

Radioprotection Effects and the Anti-Cancer Effects of the Species (Miroku-Tea)

Gu, Yeun-Hwa,Suzuki, Ikukatu,Hasegawa, Takeo,Hayashi, Ikuo,Hitoshi, Tawaraya,Ahn, Kyoo-Seok INSTITUTE OF ORIENTAL MEDICINE KYUNG-HEE UNIVERSIT 2001 JOURNAL OF ORIENTAL MEDICINE Vol.6 No.1

In recent years the effectiveness of species (Miroku-Tea) as an anti-cancer, anti0biotic effect and anti-virus effect has become more widely known. Furthermore the beneficial effects of species (Miroku-Tea) leaves have also been studied. In this study, we investigated measurement of blood cells, the NK cell activity, the radio-protective and anti-cancer effects of species (Miroku-Tea) extract. The leucocytes and the lymphocyte were transietnly increased after injection of species (Miroku-Tea). Anti-tumor effect was occurred after the injuection of species (Miroku-Tea). NK cells activity increased 2 times after the injection of species (Miroku-Tea) compared with the control group. Radiation protection occurred from the inhibition of body weight loss and also a prolonged the surviving fraction after the injection of species (Miroku-Tea).

A study of the radioprotection effect of guarana (Paullinia cupana) on the fetuses of ICR mice THE RADIATION PROTECTION EFFECTS OF GUARANA

Gu, Yeun-Hwa,Hasegawa, Takeo,Suzuki, Ikukatsu,Yamamoto, Youichi,Yoon, Yeog-Byung,Rhee, Soo-Yong Korean Association for Radiation Protection 2001 방사선방어학회지 Vol.26 No.4

Guarana, a tropical plant is found in powdered for in health food and is very popular soft drink in Brazil as an energy feaster with its high caffeine contents. We examined its radioprotection effects during organogenesis stages of ICR mice by malformations rate and cellular lead 8 the embryo by radiation and analyzed the mechanism of the radioprotection effects in the fetal of ICR mice. The results of this study showed that Guarana reduced clearly the embryonic death rate and teratogenesis rate by radiation. Its radioprotection effect inject be related with its radioprotection effect might be related with its antioxidant effect or free radical scavenger. We need to exposure the Guarana as a potential radioprotection agent. Therefore, we investigated about radiation effects by Guarana using to mice experiments in this paper.