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Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity

  • Oh, Im-Kyung;Yoo, Sang-Ho;Bae, In-Young;Cha, Jae-Ho;Lee, Hyeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.985-990
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    • 2004
  • Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

Single-dose Oral Toxicity Study of β-glucosidase 1 (AtBG1) Protein Introduced into Genetically Modified Rapeseed (Brassica napus L.) (GM 유채에 도입된 β-glucosidase 1 (AtBG1)의 단회투여독성시험)

  • Lee, Soonbong;Jeong, Kwangju;Jang, Kyung-Min;Kim, Sung-Gun;Park, Jung-Ho;Kim, Shinje
    • Journal of Life Science
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    • v.27 no.2
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    • pp.194-201
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    • 2017
  • Rapeseed (Brassica napus L.) is an oil crop classified as Brassicaceae, and it is widely grown worldwide. To develop a drought-resistant rapeseed, the ${\beta}$-glucosidase 1 (AtBG1) gene was introduced into rapeseed because drought- and salt-resistance phenotypes were observed when the AtBG1 gene was overexpressed in arabidopsis. Newly developed genetically modified crop must be proved to be safe. Safety assessments are based on the historical usage and scientific reports of a crop. In this study, we examined the potential acute oral toxicity of AtBG1 protein expressed in genetically modified (GM) rapeseed and calculated the minimum lethal dose at 6 weeks in both male and female ICR mice. AtBG1 protein was fed at a dose of 2,000 mg/kg body weight in five male and five female mice according to the marginal capacity concentration of OECD, 2,000 mg/15 ml/kg. Mortalities, clinical findings, and body weight changes were monitored for 14 days after dosing, and postmortem necropsy was performed on day 14. This study showed that no deaths occurred in the test group, and AtBG1 protein did not result in variations in common symptoms, body weight, and postmortem findings between the two groups. This showed that the minimum lethal dose of AtBG1 protein expressed in transgenic rapeseed exceed 2,000 mg/kg body weight in both sexes.

Studies on the Improvements of Functional Properties of Sardine Protein by Plastein Reaction -2. General Properties of Plasteins- (Plastein반응을 이용한 정어리 단백질의 기능성 개선에 관한 연구 -2. Plastein의 일반적 성장-)

  • Kim, Se-Kwon;Kwak, Dong-Chae;Cho, Duck-Jae;Lee, Eung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.3
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    • pp.242-248
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    • 1988
  • Plasteins were synthesized from a peptic sardine protein hydrolysate by pepsin, ${\alpha}-chymotrypsin$ pretense(from Aspergillus saitoi) and papain under the optimum conditions of previous paper. L -glutamic acid diethylester and L-leucine ethylester also were incorporated into plastein during the plastein reaction by papain. General composition, yield, molecular weight and amino acid composition were measured. The protein, ash and lipid rontent of plasteins were $81.1{\sim}88.2%$, $1.9{\sim}7.6%$ and $0.3{\sim}0.8%$, respectively. The yield of plasteins were pretense plastein 52.3%, papain plastein 44.2%, pepsin plnstein 43.6%, ${\alpha}-chymotrypsin$ plastein 43.2%. Leu -papain plastein 33. 2% and Glu - papain plastein 29.0%. The glutamic acid and leucine content in Glu -papain plastein and Leu -papain plastein were 39.0%, 37.5%, respectively. While the contents in the papain plastein were 14.3%, 7.1%, respectively. The amino acid composition of plasteins were similar to that of peptic sardine protein hydrolysate. The major molecular weight of the peptic hydrolysnte estimated by gelfilteration were 1,800 and 285, and those of plasteins were 26,000 and 9,100 for ${\alpha}-chymotrypsin$, 23,000, 10,000 and 4,300 for pepsin, 18,000 for pretense, 13,000 for papain, 29,000 for Leu -papain plastein and 19,000 for Glu -papain plastein.

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The Severity of the Pediatric Patients admitted at NICU using Therapeutic Intervention Scoring System (환자분류체계를 이용한 NICU 입원 환아의 중증도)

  • Kim, Moon-Sil;Moon, Sun-Young;Lee, Kyoung-Sook;Jeong, Yu-Kyoung;Kim, Shin-Jeong
    • Journal of Korean Academy of Nursing Administration
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    • v.8 no.1
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    • pp.5-15
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    • 2002
  • This study was attempted to help in explore new direction about classification of the severity of the pediatric patients admitted at NICU. Data were collected from 230 patients who admitted at Neonatal Intensive Care Unit of 3 University hospitals and 1 General hospital during 7 months period from september 1, 2000 to April 30, 2001. The results were as follows: 1. The degree of severity of the pediatric patients admitted at NICU shown ranged 1-102 and averaged 17.7. 2. With the respect to the severity of the pediatric patients admitted at NICU, there were statistically significant relation in passing day(s) to admission(r=-.153, p=.020), hospital day(s)(r-.501, p=.000), gestational age(r=-.354, p=.000), birth weight(r=-.280, p=.000), Apgar score at 1 min and at 5 min(4=-.340, p=.000; r=-.322, p=.000), present body weight(r=-.151, p=.023). 3. The severity of the pediatric patients according to general characteristics, there were significant difference in admitting day of the patients(t=2.339, p=.020), Apgar score at 1min and 5min(F=7.893, p=.000; t=3.568, p=.001).

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Subchronic Oral Toxicity Study of Acanthopanax divaricatus var. albeofructus in Rats

  • Kim, Myoung Jun;Lee, Mi Ju;Lee, Yong-Hoon;Park, Sun Hee;Kim, Duyeol;Park, Cheol Beom;Kang, Jin Seok;Kang, Jong-Koo
    • Toxicological Research
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    • v.33 no.1
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    • pp.15-23
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    • 2017
  • Acanthopanax divaricatus (Siebold & Zucc.) Seem. var. albeofructus (ADA), a traditional medical herb, has been used to treat arthritis and muscular injury, to strengthen muscle and bone, and to get vital energy. However, information regarding its toxicity is limited. ADA was administered by oral gavage to groups of rats at doses of 0 (control), 1,000, 1,500, 2,000, 2,500, and 3,000 mg/kg five times per week for 13 weeks. Mortality, clinical signs, body weights, food consumption, hematology, serum chemistry, urinalysis, organ weights, necropsy, histopathological finding, vaginal cytology, and sperm morphology were compared between control and ADA-treated groups. Salivation was intermittently observed in both sexes receiving 2,500 and 3,000 mg/kg directly after dosing. Absolute liver weights increased in females receiving 2,000, 2,500, and 3,000 mg/kg ADA (P < 0.05, P < 0.01, and P < 0.01, respectively) and so did the relative liver weights (P < 0.001). Salivation and increased liver weight were ADA-related changes but not considered to be adverse effects. Salivation was intermittent and transient, and the liver weight increase was minor and not accompanied by other changes such as hepatic morphological or functional alterations. The no-observed-adverse-effect-level was determined to be at least 3,000 mg/kg in both sexes of rats.

Mouse Single Oral Dose Toxicity Test of Lonicerae Flos Aqueous Extracts (금은화(金銀花)추출물의 마우스 단회 경구투여 독성실험)

  • Yoo, Hyo-Jeong;Park, Mee-Yeon;Choi, Hae-Yun;Kim, Jong-Dae
    • The Journal of Internal Korean Medicine
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    • v.31 no.3
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    • pp.539-553
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    • 2010
  • Objectives : The object of this study was to obtain accurate information (single oral dose toxicity) of Lonicerae Flos (LF; Dried flower bud parts of Lonicera japonica Thunb (Caprifoliaceae)), which has traditionally been used in Korean medicine for treating various inflammatory diseases. Methods : In order to observe the 50% lethal dose (LD 50), approximate lethal dosage (ALD) and target organs, test articles were once orally administered to female and male ICR mice at dose levels of 2,000, 1,000, 500 and 0 (control) mg/kg (body weight.). The mortality and changes on body weight, clinical signs and gross observation were monitored for 14 days after single oral treatment of LF aqueous extracts with organ weights and histopathological observations of 12 types of principle organs. Results : 1. After single oral treatment of LF aqueous extracts, we could not find any mortality and toxicological evidences up to 2,000 mg/kg treated group, the limited dosages in rodents at body and organ weights, clinical signs, gross and histopathological observations. 2. Slight diarrhea was detected in most mice treated with 2,000 mg/kg of LF aqueous extracts and male mice of LF aqueous extracts 1,000 mg/kg within 2 days after end of treatment, respectively. Conclusion : The results obtained in this study suggest that the LD 50 and ALD of LF aqueous extracts in both female and male mice after single oral treatment were considered as over 2,000 mg/kg because no mortalities were detected up to 2000 mg/kg, the highest dose recommended by KFDA and OECD. However, we also observed the possibility of digestive disorders like diarrhea when over 1,000 mg/kg of LF aqueous extracts were administered in the present study.

The effects of vitamin A supplementation during late-stage pregnancy on longissimus dorsi muscle tissue development, birth traits, and growth performance in postnatal Korean native calves

  • Jo, Yong Ho;Peng, Dong Qiao;Kim, Won Seob;Kim, Seong Jin;Kim, Na Yeon;Kim, Sung Hak;Nejad, Jalil Ghassemi;Lee, Jae Sung;Lee, Hong Gu
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.5
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    • pp.742-752
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    • 2020
  • Objective: This study investigated the effects of vitamin A (VA) supplementation during late-stage pregnancy on longissimus dorsi muscle tissue development, birth traits, and growth performance of postnatal Korean native calves. Methods: In the preliminary experiment, twenty-six pregnant cattle (initial body weight [BW] = 319 kg (standard deviation [SD] = 30.1; 1st parity) were randomly assigned to the control and treatment groups. The treatment group received VA supplementation at 24,000 IU/d from gestational day 225 until delivery. In the main experiment, twelve pregnant cattle (initial BW = 317 kg [SD = 31.3]; 1st parity) were treated with VA supplementation at 24,000 IU/d (gestational days 150 to 225) and at 78,000 IU/d (gestational day 225 until delivery). Serum VA levels were analyzed in pregnant cattle, and the growth performance, gene expression, and serum VA levels were analyzed in the offspring. Results: Serum VA levels in pregnant cattle decreased the late gestation in both experiments (p<0.001). In the main experiment, pregnant cattle at parturition and offspring at birth in the treatment group had higher serum VA levels than those in the control group (p<0.05). In the treatment groups, an increased birth weight was observed in the main experimental group (p = 0.022), and a tendency (p = 0.088) toward an increased birth weight was observed in the preliminary experimental group. However, no differences were observed in the feed intake, average daily gain, gain-to-feed ratio, or BW of 31-day-old calves. Gene expression was analyzed in longissimus dorsi muscles of 31-day-old calves. VA supplementation in pregnant cattle stimulated postnatal muscle development in offspring by elevating myogenic factor 5 (MYF5), MYF6, and myoblast determination levels (p<0.05). Moreover, preadipocyte-related marker genes such as extracellular signal-regulated kinase 2 and krüppel-like factor 2 were higher in the treatment group than in the control group (p<0.05). Conclusion: VA supplementation (78,000 IU/d) in late-stage pregnant cattle maintained serum VA levels. In addition, 78,000 IU/d VA supplementation increased the birth weight and expression of genes related to muscle and preadipocyte development in offspring. Overall, 78,000 IU/d VA supplementation in pregnant cattle is beneficial to newborn calves.

Studies on hydrophobic drug-soluble carrier coprecitates 2

  • Shin, Sang-Chul
    • Archives of Pharmacal Research
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    • v.2 no.1
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    • pp.49-64
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    • 1979
  • In an atempt to elucidate further physicochemical properties of furosemide-PVP coprecipitates, extensive investigations such as TLC, UV,IR, NMR, X-ray diffraction, TGA and DTA studies were carried out for the furosemide test systems. X-ray diffraction studies revealed that the pure furosemide and the furosemide contained within a physical mixture were crystalline in nature. However, there was no crystallinity evident in the 1:5 furosemide-PVP 40,000 coprecipitate system, even after standing for two years. The various ratio furosemide-PVP 40,000 coprecipitate systems revealed that the coprecipitate containing a greater amount of PVP 40,000 than that of furosemide showed a crystalline state of furosemide and that the minimum amounts of PVP to make amorphous form of furosemide was 1:1 ratio of furosemide to PVP. From the furosemide-PVP coprecipitate systems with PVP of different molecular weights of 10,000, 40,000 and 360,000, all the 1:1 ratio coprecipitates did not exhibit any crystallinity of furosemide, whereas all the 2:1 ratio coprecipitates showed a presence of crystalline furosemide. All the coprecipitated preparations with PEG 4,000 and with PEG 6,000 showed the diffraction peaks indicating the presence of crystalline furosemide. The comparison of infrared spectra of the physical mixture and the coprecipitate showed an interaction such as association between the functional groups of furosemide and PVP in the molecular level, whereas the studies by TLC, UV and NMR showed its dissociation in methanol solution. The weight losses in TGA curves showed all the same patterns. However, a little different transition form in DTA thermograms was shown between the physical mixture and the coprecipitate, indicating the different thermal property.

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Four-Week Repeated Oral Toxicity Study of Clean Natural in Sprague-Dawley Rats (Clean Natural의 Sprague-Dawley 랫드를 이용한 4주 반복경구투여 독성시험)

  • Kim, Eui-Kyung;Kim, Jun-Young;Lee, Hu-Jang
    • Toxicological Research
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    • v.21 no.3
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    • pp.263-269
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    • 2005
  • This study was performed to investigate repeated-dose toxicities of Clean natural, a new disinfectant, in Sprague-Dawley(SD) rats. In the 4-week repeated oral toxicity study, Clean Natural was orally administered once daily via gavage to male and female rats at dose levels of 0, 500, 1,000 and 2,000 mg/kg body weight for 4-weeks. There were no deaths and clinical signs during the dosing period. In both sexes, there were no statistically significant differences between the administered and control groups in urinalysis indicators and hematological parameters. In serum biochemistry, aspartate aminotransferase(AST) was significantly decreased and sodium content was increased in the 2,000 mg/kg male group, while chlorine was significantly decreased in the 2,000 mg/kg female group. Also, albumin, total cholesterol and total bilirubin were significantly increased in the 2,000 mg/kg male and female group. In histopathological examinations, centrilobular hepatocellular hypertrophy in the liver was observed in the 2,000 mg/kg male and female groups. And pigmentation in the spleen was observed in the 2,000 mg/kg male group. In conclusion, four-week repeated oral dose of Clean Natural to rats did not cause apparent toxicological change at the dose less than 2,000 mg/kg body weight. Thus it is suggested that no-observed adverse-effect level(NOAEL) for Clean Natural in rats was considered to be 1,000 mg/kg/day.

Changes in the Pectic Substance during Ripening of Salted Cucumber Pickle (염지 오이피클의 숙성중 펙틴질의 변화)

  • 오영애;이만정;김순동
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.2
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    • pp.143-150
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    • 1990
  • The changesin hardness activities of pectinestrase and polygaacturonase and amounts of pectic substances of cucumber during salting at 1$0^{\circ}C$ were investigated, The hardness of the cucumber was decreased dramatically after 3 weeks whereas activities of pectinestrase and polygalacturonase were increased until 3 weeks and 2 weeks respectively and then decrea-sed. The level of alcohol insoluble solid and protopectin in the cucumber were decreased but those of pectic acid and water soluble pectin were increased during the whole salting periods, Protopectin fractionated from alcohol insoluble solid during salting of cucumber was separated using Sephacryl S-500 It showed that high average molecular weight(AMW) of 100,000 was decreased however lower molecular weight compounds was increased. Pectic acid was observed to be decomposed from AMW 200,000 to AMW 500,000 Water soluble pectin from fresh cucumber contained higher level of pentose with peak I of AMW 2,000,000, however after 6 weeks of saltinf peak II which represented AMW 100,000 was separated. From the changes of sugar composition, the phenomena of softness during the salting was probably caused by solubilization of hemicellulose associated with pectin and decomposition of pectic substances.

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