• Title/Summary/Keyword: LD$^{50}$

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Mucosal Administration of Lactobacillus casei Surface-Displayed HA1 Induces Protective Immune Responses against Avian Influenza A Virus in Mice

  • Dung T. Huynh;W.A. Gayan Chathuranga;Kiramage Chathuranga;Jong-Soo Lee;Chul-Joong Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.735-745
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    • 2024
  • Avian influenza is a serious threat to both public health and the poultry industry worldwide. This respiratory virus can be combated by eliciting robust immune responses at the site of infection through mucosal immunization. Recombinant probiotics, specifically lactic acid bacteria, are safe and effective carriers for mucosal vaccines. In this study, we engineered recombinant fusion protein by fusing the hemagglutinin 1 (HA1) subunit of the A/Aquatic bird/Korea/W81/2005 (H5N2) with the Bacillus subtilis poly γ-glutamic acid synthetase A (pgsA) at the surface of Lactobacillus casei (pgsA-HA1/L. casei). Using subcellular fractionation and flow cytometry we confirmed the surface localization of this fusion protein. Mucosal administration of pgsA-HA1/L. casei in mice resulted in significant levels of HA1-specific serum IgG, mucosal IgA and neutralizing antibodies against the H5N2 virus. Additionally, pgsA-HA1/L. casei-induced systemic and local cell-mediated immune responses specific to HA1, as evidenced by an increased number of IFN-γ and IL-4 secreting cells in the spleens and higher levels of IL-4 in the local lymphocyte supernatants. Finally, mice inoculated with pgsA-HA1/L. casei were protected against a 10LD50 dose of the homologous mouse-adapted H5N2 virus. These results suggest that mucosal immunization with L. casei displaying HA1 on its surface could be a potential strategy for developing a mucosal vaccine against other H5 subtype viruses.

Repellent and Acaricidal Activities against Leptotrombidium pallidum Larvae of Eucalyptus Oil (털진드기 유충에 대한 유칼립투스 오일의 기피 및 살비활성)

  • Jo, Hyeong-Chan;Kim, Kwang-Ho;Lee, Sang-Guei;Na, Young-Eun;Park, Hyung-Man
    • Korean journal of applied entomology
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    • v.47 no.3
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    • pp.287-292
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    • 2008
  • Repellent and acaricidal activities of eucalyptus oil, permethrin, and DEET against Leptotrombidium pallidum larvae, which are a vector transmitting tsutsugamushi disease, were evaluated under laboratory conditions using a filter paper impregnated method. The $LD_{50}$ values of eucalyptus oil and DEET were 0.025 and 0.018 $mg/cm^2$, respectively while that of permethrin was higher than 0.2 $mg/cm^2$. In the repellency test of these materials at 6.14 $mg/cm^2$, eucalyptus oil gave complete repellency, and the larvae crossed the treated zone killed. But permethrin showed 60% repellency at 9.20 $mg/cm^2$ and the mites croosed the zone were not killed. The percent repellency of DEET at 0.53 $mg/cm^2$ was 8.3 and 2.8 times higher than that of permethrin and eucalyptus oil, respectively. The acaricidal activities of emulsifiable concentrates-pump sprayers containing the eucalyptus oil as an active ingredient were assayed. The emulsifiable concentrates containing 1% and 3% eucalyptus oil showed weak mortality at 1 hour after treatment, while all ones containing more than 6% oil produced 100% activity against L. pallidum larvae. The mortality also increased as exposure time to the concentrates increase. These results suggest that the potential of eucalyptus oil highly expected to be used as a control or repellent agent against L. pallidum larvae may be very high.

Single- and Repeated-Dose Oral Toxicity in Rats and Bacterial Reverse Mutation Test of Morus alba L. Extracts (상지추출물의 단회/반복투여 독성 및 복귀돌연변이능 평가)

  • Han, Taewon;Um, Min Young;Lim, Young Hee;Kim, Jeong-Keun;Kim, In-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.10
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    • pp.1406-1413
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    • 2016
  • This study was carried out to evaluate the toxicity of ethanolic extracts of Morus alba L. branch (ME). In the reverse mutation test, Salmonella Typhimurium TA98, TA100, TA1535, TA1357, and Escherichia coli WP2uvrA were used to estimate the mutagenic potential of ME. Sprague-Dawley rats were orally administered ME at levels of 1,250, 2,500, and 5,000 mg/kg for the single-dose toxicity test and 500, 1,000, and 2,000 mg/kg/d for the repeated-dose toxicity test for 28 consecutive days. As expected, reverse mutation was not detected at any concentration of ME, regardless of application of the metabolic activation system with or without S9 mix. In the single-dose toxicity test, ME caused neither significant visible signs of toxicity nor mortality in rats, and $LD_{50}$ was estimated to be over 5,000 mg/kg. In the repeated-dose toxicity test, ME administration at 500, 1,000, and 2,000 mg/kg for 28 days to male or female rats did not result in mortality. Similarly, no toxicologically significant treatment-related changes in body weight, food intake, or organ weights were noted. Several hematological and biochemical parameters in both genders showed significant differences, but these were within normal ranges. These results support the safe use of ME.

Effect of phenobarbital sodium and 3-methylcholanthrene on metabolism in vitro and toxicity of $^{14}C$-carbofuran in rat (쥐에서 phenobarbital sodium 및 3-methylcholanthrene이 $^{14}C$-carbofuran의 독성과 in vitro 대사에 미치는 영향)

  • Han, Seong-Soo;Rim, Yo-Sup
    • The Korean Journal of Pesticide Science
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    • v.2 no.2
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    • pp.29-38
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    • 1998
  • In order to elucidate the effect of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on metabolism in vitro and toxicity of $^{14}C$-carbofuran in rat, they were administered by the chemicals, alone or in combination, and their survival ratios and metabolites were investigated. The $LD_{50}$(96 hrs) value of carbofuran to rats was 6.9 mg/kg. The toxicities of the major metabolites were in the decreasing order of 3-hydroxycarbofuran, 3-ketocarbofuran, 3-hydroxycarbofuran phenol and were much lower than that of the parent compound. When the rats were orally administered by the dose of carbofuran alone, 8.4 mg/kg, the survival ratio was 0%, whereas that was raised up to $60{\sim}80%$ with 20 mg/kg of PB or 3-MC, and 100% with 60 mg/kg of PB or 3-MC. Their metabolism in vitro occurred in the microsomal fraction. In case of carbofuran alone, the major metabolite was 3-hydroxycarbofuran. When carbofuran with PB or 3-MC, on the other hand, was treated, it was 3-ketocarbofuran. In addition, when the co-factor(NADP+G-6-P+G-6-P-DG) was added to the microsomal fraction(phase I system), and a mixture of NADPH+GSH to the 105,000g supernatant(phase II system) taken by carbofuran alone, each metabolites were produced by the maximum levels, respectively. In case of the carbofuran treatment with PB or 3-MC, the microsomal fraction of phase I system produced the maximum levels of metabolites, as in the treatment of carbofuran alone, whereas the 105,000g supernatant supplemented with the co-factor NADPH+FAD(phase II system) was brought about the maximum production of metabolites. The ratio of the formation of metabolites was 2 to 3 times higher in the combined treatment of carbofuran with PB or 3-MC than in the treatment of carbofuran alone.

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Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae) (담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성)

  • Kim, Da-A;Kim, Jin-Su;Kil, Mi-Ra;Paek, Seung-Kyoung;Choi, Su-Yeon;Jin, Da-Yong;Youn, Young-Nam;Hwang, In-Cheon;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.47 no.1
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    • pp.87-93
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    • 2008
  • Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.

Study of Kidney Toxicity of Azadirachta Indica Extract for Oral Administration in Rats (님추출물의 경구투여에 따른 랫드의 신장독성 연구)

  • Yoon, Hyunjoo;Choe, Miseon;Cho, Hyeon-Jo;Han, Beom Seok;Park, Kyung-Hun;Oh, Jin-Ah;Cho, Namjun;Paik, Min-Kyoung
    • Korean Journal of Environmental Agriculture
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    • v.33 no.2
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    • pp.103-110
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    • 2014
  • BACKGROUND: Azadirachta indica has been widely used as environment-friendly organic materials because of its insecticidal properties. This study was carried out to investigate the acute toxicity and the subacute toxicity of Azadirachta indica extract(AIE) in rats. METHODS AND RESULTS: For the oral acute toxicity test, Sprague-Dawley rats were gavaged with 2.0 g/Kg bw of AIE. The $LD_{50}$ value was greater than 2.0 g/Kg bw for both male and female rats. For the subacute toxicity study, rats were treated with AIE at doses of 0.5, 1.0, 2.0 mg/Kg bw once a day for 4 weeks(n=10 animals per each group). There were no significant changes in body weight, food intake and water consumption observed during the experimental duration. In addition, no difference of relative kidney weight was observed among all treated groups. Serum creatinine level in the AIE 2.0 g/Kg group increased significantly compared with that of control group in male rats, but serum blood urea nitrogen was significantly decreased in a dose-dependent manner (p<0.05). Significant increase of serum cholesterol levels were observed in all AIE groups, compared with the control group, in the female rats (p<0.05). However, histopathological examination of the kidney did not reveal any significant lesions in all groups. CONCLUSION: On the basis of results, it could be concluded that oral administration AIE didn't cause any toxic response in kidney, except the increased serum cholesterol.

Acute Oral Toxicity and Anti-obesity Effect of Diglyceride Preparation Containing Conjugated Linoleic Acid in Rat (공액리놀레산 함유 디글리세라이드 식용유지 조성물의 rat에 대한 단회 경구투여독성 및 항비만 효과)

  • Hong, Soon-Gi;Park, Chae-Kyu;Lee, Mi-Ja;Chung, Shin-Gyo;Lee, Young-Ho;Hyun, Sun-Hee
    • Korean Journal of Food Science and Technology
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    • v.41 no.3
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    • pp.320-325
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    • 2009
  • The present study was carried out to investigate the acute oral toxicity and anti-obesity effects of a diglyceride preparation containing conjugated linoleic acid (DG+CLA). To test its acute oral toxicity, the DG+CLA was injected into 30 rats (15 males and 15 females) at dosage of 2,000 mg/kg and 5,000 mg/kg. Mortality rates, clinical signs, and body weight changes were monitored for 14 days following administration. According to the results, the lethal dose ($LD_50$) of DG+CLA was determined as >5,000 mg/kg in both sexes. There were no significant changes in general conditions, clinical signs, body weight, and gross lesions between the vehicle control and DG+CLA groups. For the anti-obesity studies, obese Zucker rats were randomly divided into 4 groups and fed saline, soybean oil, diglyceride, and DG+CLA, respectively, for 8 weeks. The DG+CLA groups presented significant differences in body weight, food efficiency ratio, serum lipid levels, and fat weight. Overall, the results showed that the DG+CLA did not have acute oral toxicity and reduced body weight, serum lipid levels, and fat gain.

Single-Dose Oral Toxicity in Rat and Bacterial Reverse Mutation Assay of Psoralea corylifolia L. Extracts (파고지 추출물의 렛트에 대한 단회 경구 투여 독성 및 복귀돌연변이능 평가)

  • Kim, Sun-A;Lim, Sun-Hye;Ahn, Ji-Yun;Kim, Sung-Ran;Ha, Tae-Youl
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.960-964
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    • 2007
  • This study was performed to examine the toxicity of Psoralea corylifolia L. by the single-dose oral toxicity tests in rat and bacterial reverse mutation assay. In single-dose oral toxicity tests, 5 mL ethanol extract of P. corylifolia L. were directly injected into 10 rats (5 males and 5 females) at a dosage of 2 g/kg. Death practice was not detected during breeding periods (14 days), and $LD_{50}$ was calculated over 2 g/kg. No difference were observed with control group in the growth rate and histological observations. In bacterial reverse mutation assay, his(-) Salmonella Typhimurium TA98, TA100, TA1535, TA1537 and trp(-) Escherichia coli WP2uvrA (pKM101) were used for assessing the toxicity of ethanol extracts of P. corylifolia L.. No significant difference in formation of the colonies and no dose-dependent increase was observed regardless of the addition of S9 mix. The results showed that ethanol extracts of P. corylifolia L. did not have single-dose oral toxicity and mutagenic toxicity.

Bioactive Characterization of Bacillus thuriniensis subsp. kurstaki CAB133 Isolated from Domestic Soil (국내 토양으로 분리된 Bacillus thuriniensis subsp. kurstaki CAB133균주의 생물학적 특성)

  • Choi, Su-Yeon;Cho, Min-Su;Kim, Tae-Hwan;Kim, Jin-Su;Pack, Seung-Kyung;Youn, Young-Nam;Hong, Soon-Sung;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.47 no.2
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    • pp.175-184
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    • 2008
  • To screen highly active Bacillus thuringiensis isolates against Spodoptera litura (Lepidoptera, Noctuidae), 46 B. thuringiensis was isolated from 115 samples obtained from several crop soils. Especially, B. thuringiensis subsp. kurstaki CAB133 and CAB162 isolates showed 100% mortality against S. litura. $LD_{50}$ values of CAB 133, CAB162 and HD-1 strains of B. thuringiensis subsp. kurstaki were 0.089, 3.144 and $0.513{\mu}g/ml$ against 2nd larva of S. litura, respectively. The weight of 3rd larva of S. litura which were fed crystal inclusion protein $(1.267{\mu}g/ml)$ with B. thuringiensis subsp. kurstaki CAB133 was about 30 times lass than control group. CAB133 and CAB 162 strains of B. thuringiensis subsp. kurstaki which were taken a highly toxity against S. litura were analyzed by SDS-PAGE, and estimated the molecular weight of the Cry proteins. Their serological identification by H serotypes were showed B. thuringiensis subsp. kurstaki (3abc) type.

Fine Structural Changes of the Renal Corpuscle of the Mice following the Administration of 5-Fluorouracil or Mitomycin C (5-Fluorouracil 및 Mitomycin C 투여후 생쥐 콩팥소체의 미세구조 변화)

  • Ko, Jeong-Sik;Oh, Won-Young;Kim, Jin-Gook;Park, Kyung-Ho;Ahn, E-Tay
    • Applied Microscopy
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    • v.29 no.1
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    • pp.25-41
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    • 1999
  • The experiment was performed to study the morphological responses of the renal glomeruli of the mice after administration of 5-fluorouracil or mitomycin C. 5-fluorouracil (60 mg/kg) or mitomycin-C $(400{\mu}g/kg)$ were injected subcutaneously to the animals every other day, and animals were sacrificed at 4 days or 7 days following the first injections. Pieces of tissues were observed with a JEM 100CX-II electron microscope. The observed results were as follows: 1. In the fourth day following the first injection of 5-fluorouracil or mitomycin C, components of the renal glomeruli of the mice are looked compact since they were filled with the widened the mesangium, and showed narrowing lumen of glomerular capillaries and of urinary spaces. The changes were more significant in the mitomycin C treated mice. 2. In the 5-fluorouracil treated mice, morphological changes of glomeruli were generally recovered in the seventh day, whereas the glomeruli of the mitomycin C treated mice have not shown general recovery. 3. In the fourth and seventh days following the first injection of mitomycin C, in the renal glomeruli of the mice, swollen endothelial cells, and protruded mesangeal cells into the capillary lumen are frequently observed. 4. In the fourth day following the first injection of mitomycin C, in the glomerular basal lamina of the mice, the electron densities of the lamina rara interna and the lamina rara externa were similar to the density of the lamina densa and the expanded lamina rara interna were often seen. From the above results, it is suggested that the cytotoxic effects of the mitomycin C on renal glomeruli are more severe as compared with those of 5-fluorouracil.

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