• Title/Summary/Keyword: In vivo

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Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

The Efficacy Evaluation of Tourmaline-Ionized Water in Animal Study (투어마린이온활성수의 효능 평가)

  • Yoon, Yang-Suk;Kim, Dong-Heui;Qi, Xu-Feng;Song, Soon-Bong;Jung, Jong-Ho;Joo, Kyung-Bok;Teng, Yung-Chien;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.39 no.4
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    • pp.311-317
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    • 2009
  • This study was performed using animals to confirm the effect of tourmaline-ionized water (TIW) the properties of which were changed by tourmaline energy and electric discharge. In the ICR mice fed high-fat diet, body weight increasing rate of the TIW-treated group (Exp) was generally decreased and moreover exhibited significance at 11th week (P<0.05) compared with the control (Con) group fed distilled water, although water intake of the Exp group was lower than that of the Con group. In the ICR mice with $CCl_4$-induced hepatotoxicity, AST and ALT activities of the Exp group were not significant but showed some decreasing trend, and histological damage of liver was less compared with thatof the Con group. On the study of ethanol-induced hangovers in Sprague-Dawley rat, blood alcohol concentration was significantly decreased (P<0.01), activity of GST, antioxidant enzyme related to the alcohol metabolism, was increased in liver tissue (P<0.05), and AST and ALT show a tendency to be decreasedin the Exp group. These results suggest that drinking TIWhas not only some obesity preventing effect but also an alcohol detoxification effect and liver protecting effect in vivo. It is supposed due to a structural change of water cluster and a property which maintains the changed structure through tourmaline energy and electric discharge. Therefore, TIW has a potentiality to be developed as functional water with several beneficial effects as well as for daily drinking, but further study on the mechanism related with efficacy will be necessary.

Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.576-587
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    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.

Sow Transfer of Cultured Freezing Embryos by Open Pulled Straw(OPS) Methods : Preliminary Results (Open Pulled Straw(OPS) 방법에 의한 체외배양 동결수정란의 경산돈 이식 : 예비실험 결과)

  • Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.155-163
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    • 2004
  • The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.

The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model (백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향)

  • Nam, Jeong-Hun;Noh, Kyung-Lok;Pang, Eun-O;Yu, Woo-Geun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Seong-Gon;Park, Young-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.366-374
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    • 2010
  • Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

Comparison of Flavonoid Contents and Antioxidant Activity of Yuzu (Citrus junos Sieb. ex Tanaka) Based on Harvest Time (산지별 유자의 수확시기에 따른 플라보노이드 함량 및 항산화활성 비교)

  • Moon, So Hyun;Assefa, Awraris Derbie;Ko, Eun Young;Park, Se Won
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.283-291
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    • 2015
  • The aim of this study was to evaluate the changes of fruit quality, flavonoid contents and antioxidant activity of Yuzu (Citrus junos Sieb. ex Tanaka) from Go-heung and Jeju according to harvest time. Samples were harvested from August to December on the $1^{st}$ of every month. August and September samples were green colored, whereas November and December samples were yellow. The fruit shape index decreased, changing from globular to elliptical, whereas the $^{\circ}birx$ increased with ripening stage. The yuzu from Jeju was larger than that from Go-heung in each month of cultivation. August samples exhibited the highest amounts of phenolic compounds. In addition, samples from Jeju had higher total phenolic content than those from Go-heung. The content of phenolic compounds decreased with ripening until October and then increased subsequently. Antioxidant activity of the yuzu was evaluated by FRAP and DPPH methods. The antioxidant activity showed a similar trend as total phenolic content. Immature yuzu fruit was found to exhibit the highest amount of flavonoids such as naringin and hesperidin. November and December samples showed almost the same contents of flavonoids. The flavonoid content of yuzu fruit harvested from Jeju was higher than that from Go-heung. Overall, the samples harvested at the early stage, in the month of August, exhibited the highest flavonoid content, phenolic content and antioxidant activity. As the health benefits of these compounds has been demonstrated in various studies, the immature yuzu appears to be preferable for use as a raw material for formulation of pharmaceutical products as well as for functional food production after a proper in-vivo and in-vitro medical tests.

Application to the Biscuits Manufacture of Processed Amaranth Seeds (아마란스(Amaranth) 종실의 가공에 따른 비스킷 제품에의 적용)

  • 김진수;유희중
    • The Korean Journal of Food And Nutrition
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    • v.15 no.4
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    • pp.321-325
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    • 2002
  • For examination, amaranth was first dried its surface after sufficient soaking in water. Dried amaranth was roasted when the moisture contents reached 30∼50%, its nasty smell disappeared and its color turned to yellow and it was puffed. Pretreated amaranth was added to biscuit for the improvement of apparence and merchandise value. The maximum expansion was reported at the moisture percentage of 130∼160% according to the examination of expansion and moisture percentage. In order to compare gelatinization of different processing, three different amaranth were prepared as follows ; Raw amaranth, steamed/dried amaranth, and roasted amaranth. The degree of gelatinization was increased as the percentage of moisture was increased and the degree of gelatinization of roasted amaranth was higher than the one of steamed/dried amaranth, their moisture contents were 62.10% and 57.59%, respectively In addition, the hardness($\times$10$\^$5/dyn/㎠) of roasted amaranth was showed lower values than that of steamed amaranth and raw amaranth were showed the large values. After examining biscuits containing each amaranth, raw amaranth had problem with nasty smell and bad texture, and streamed/dried amaranth were able to remove viscosity but turned brown. Biscuits containing roasted amaranth had good smell and texture, and besides the color of biscuits became bright because roasted amaranth turned white. Biscuits tasted best when it contained 5% of roasted amaranth. The rancidity of biscuits with 5% roasted amaranth were proceeded slowly while roasted amaranth itself had high acid value and peroxide value. For the safety from oxidation, it was fairly safe for about 6 months.

The Roles of Excitatory Amino Acid System in the Organophosphate-induced Brain Damage (유기인제에 의한 뇌 손상에 있어서 흥분성 아미노산의 역할)

  • Ko, Bong-Woo;Park, Eun-Hae;Kim, Dong-Sik;Bang, Sung-Hyun;Jin, Joo-Yeon;Kim, Dae-Sung;Ju, Chang-Wan;Lee, Kyung-Kap;Cho, Moon-Jae;Kimcho, So-Mi;Lee, Bong-Hee;Riu, Key-Zung;Park, Min-Kyoung;Lee, Young-Jae
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.148-152
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    • 2001
  • This study investigated the role of excitatory amino acid systems in the initiation of organophosphate-induced seizures and brain damages in rats through quantitative in vivo microdialysis. Microdialysates were collected from the hippocampus of rat brain, treated with diisopropylfluorophosphate (DFP; 2.67 mg/kg, s.c.) alone, and/or atropine sulfate (15 mg/kg, i.m.) and procyclidine (30 mg/kg, i.m.). The protective effects of atropine, a muscarinic blocker, and/or procyclidine, a N-methyl-D-aspartate and cholinergic antagonist, against DFP were examined. DFP treatment increased the levels of aspartate (Asp) and glutamate (Glu) significantly in the hippocampal persuate with the induction of seizures. Treatment of procyclidine could effectively block the increase of Asp and Glu levels. Atropine treatment showed no significant anticonvulsive effects against DFP-induced seizures. The increases of Asp and Glu levels by DFP were also completely blocked through the combined treatment of atropine and procyclidine. Histopathological findings on the hippocampus confirmed the above results. More effective protection was observed through the treatments of procyclidine alone or of both procyclidine and atropine than atropine alone against DFP-induced brain damage. Procyclidine was shown to be effective in DFP-induced seizures.

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Hemopoietic and Radioprotective Effects of Shenrong Fuzheng Tang(S.F.T.;蔘茸扶正湯) (삼용부정탕(蔘茸扶正湯)의 방사선(放射線) 조사(照射)로 손상(損傷)된 조직(組織) 회부(恢復) 및 조혈촉진(造血促進) 효과(效果))

  • Kim, Jeong-Su;Choe, Seung-Hun;An, Gyu-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.129-147
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    • 1997
  • Radiotherapy is an irreplaceable method of cancer treatment. But it has several side effects, especially damages to the hemopoietic and Immune system. Therefore radioprotectors are required to treat cancer successfully. A lot of Herbs and Herbal prescriptions are reported to have radioprotective effects. Above all, those to support the healthy energy and strengthen the body resistance are found more effective. This study was performed to evaluate the radioprotective effects of prescription Shenrong Fuzheng Tang(S.F.T.), which consists of 16 kinds of herbs. We investigated proliferation of murine splenocytes, secretion of colony-stimulating-factors(CSFs), immunocompetence after irradiation in-vitro, and Endogenous spleen colony assay, survival assay in-vivo. When splenocytes were cultured with Shenrong Fuzheng $Tang(S.F.T.)(500{\mu}g/ml)$, proliferation was enhanced 5.7 times compared to control cultured with medium alone(p<0.05) and, showed highest proliferation at 4th day after incubation. In order to evaluate stimulation of hemopoiesis of Shenrong Fuzheng Tang(S.F.T.), the supernatant of splenocytes cultured with optimal concentration of Shenrong Fuzheng Tang(S.F.T.) was used to measure CSFs secretion. The result showed enhanced secretion of colony-stimulating-factors (CSFs) compared to control(p<0.05). To evaluate the protective effect of lymphocytes from irradiation, proliferation of splenocytes stimulated by LPS and ConA after incubation with Shenrong Fuzheng Tang(S.F.T.) for 24h Prior to Irradiation$(1{\sim}3\;Gy)$ was measured. The results showed higher proliferation of Shenrong Fuzheng Tang(S.F.T.) treated cells than that of non-treated cells. And percentage increases of irradiated splenocytes per non-irradiated splenocytes were also higher in Shenrong Fuzheng Tang (S.F.T.)-treated cells than control. Endogenous spleen colony assay. to evaluate the protection of hemopoietic cells from irradiation, showed increased number of colonies(p=0.03) in Shenrong Fuzheng Tang(S.F.T.) treated murine spleen$(10.3{\pm}1.9)$ compared to non-treated murine spleen$(3.4{\pm}0.8)$. Survival time of mice irradiated with lethal dose of ${\gamma}-ray(9Gy)$ was prolonged in Shenrong Fuzheng Tang(S.F.T.) treated group prior to irradiation as compared to non-treated group. According to these results we can suggest that prescription Shenrong Fuzheng Tang(S.F.T.) has radioprotective effects and can be used to protect the hemopoietic and immune system from damages of anti-cancer radiotherapy.

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Inhibitory Substance on the Snake Venoms Produced by Penicillium sp. (사독의 조해물질에 관한 연구)

  • Seu, Jung-Hwn;Yi, Dong-Heui
    • Microbiology and Biotechnology Letters
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    • v.7 no.2
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    • pp.75-89
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    • 1979
  • One strain of Penicillium sp. (175-66-B), isolated from soil, was able to produce a substance that has a strong inibition activity against the Agkistrodon and Trimeresurus venoms. In this experiment, the chemical and biological properties of the sample were investigated. As an inhibitory substance, it was effective to the proteinase, hemorrhagic and lethal factors of Agkistrodon and Trimeresurus venoms, and also effective to several fractions of the proteinases and hemorrhagic factors of Agkistrodon halys blomhoffi venom. Moreover, in the addition of prednisotone, it was more effective for the cure of the mouse envenomated with the venom amount of two fold of MLD$_{100}$. This substance was very stable to the acid, alkali and heat. Its melting point was high enough to sublime at 222$^{\circ}C$ without any decomposition. This sample was easily dissolved only in hot water, but not in several organic solvents except for a little dissolution in elate. It did not have the chelating activity. It had very strong specificity to the snake venoms. but its activity was depressed by the addition of zinc or cupric salts. This sample had no acute toxicity to the mouse. Its chemical formula was $C_{16}$ $H_{12}$$N_2$ $O_{10}$ with the molecular weight of about 392. It has two epoxy groups and four carboxyl radicals, but amino, nitrite and nitrate radicals, unsaturated bonds and aromatic ring were not detected. Theuchemical configuration of this sample was suggested to be;

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