• Title/Summary/Keyword: Antiserum

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Comparative Investigation of Glutathione S-Transferases, Glyoxalase-I and Alliinase Activities in Different Vegetable Crops

  • Hossain, Md Daud;Rohman, Md Motiar;Fujita, Masayuki
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.19-26
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    • 2007
  • Glutathione S-transferases(GSTs, EC 2.5.1.18), glyoxalase-I(EC 4.4.1.5) and alliin lyase(alliinase, EC 4.4.1.4) are important enzyme systems in plant bodies. The first two are mainly detoxifying enzymes that utilize glutathione(GSH) in the defense mechanism, and the last one is mainly involved in secondary metabolism and relevant to sulfur compounds derived from GSH. The activities of the three enzymes have been investigated in soluble extracts of vegetable crops, including pumpkin, cabbage, broccoli, radish, carrot, potato, sweet potato, mungbean, and onion. GST activities were detected in all of the vegetables, and the extract of onion bulb exhibited the highest specific activity(648 nmol/min/mgP). The putative GSTs of most of the vegetables were found to be induced by ethanol. The activities of GSTs in onion bulb were found to be markedly inhibited by S-hexyl glutathione and were also inhibited by S-butyl glutathione and S-propyl glutathione. The anti-CmGSTF1 antiserum recognized a thick band for putative onion GST. The estimated glyoxalase-I activity level was also high in onion bulb(4540 nmol/min/mgP), indicating that the thick band detected by Western blot analysis might result from partial recognition of glyoxalase-I by the antiserum. The specific activities for glyoxalase-I were moderate in radish and carrot, and the extracts of other vegetables had rather low levels of activities. The extract of onion also showed the highest specific activity level for alliinase(2069nmol pyruvate/mgP). The extracts of other vegetables also had alliinase activities, although the estimated values were much lower than that of onion.

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Study on the Distribution of Vibrio parahaemolyticus from Various Kinds of Shells in Kunsan Bay (군산만에서 서식하는 패류의 장염 비브리오에 관한 분포연구)

  • 윤한식;안병용
    • Journal of Food Hygiene and Safety
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    • v.7 no.4
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    • pp.137-142
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    • 1992
  • The distribution of V. parahaemolyticus was surveyed from various kinds of shells in Kunsan Bay from July to September, 1987. 1be morphological, biochemical and serological characteristics of the isolated strains were studied. The results were as follows: 1. 41 strains were isolated from 1,350 specimens of shells (Crassostrea gigas, Tapes philippinarum, Meretrix Iusoria) 2. The isolation rates of V. parahaemolytieus were 3% in July, 3.8% in August, and 2.2% in September, respectively. 3. V. parahaemolytkus was more frequently isolated from Kunsan (20 strains) than Bideukgi (12 strains) and Gae Hwa-do (9 strains). 4. V. parahaemolyticus was more frequently isolated from C gigas (23 strains) than other shells. 5. Kanagawa hemolysis reactions were all negative. 6. 6 Strains positive to K pooled antiserum included K IV, K V, K VI and K vn type.

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Reproductive fecundity of Iraqi Awassi ewes immunized against synthetic inhibin-α subunit or steroid-free bovine follicular fluid

  • Al-Sa'aidi, Jabbar Abbas Ahmed;Khudair, Khalisa Khadim;Khafaji, Sura Safi
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1169-1175
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    • 2018
  • Objective: The present study was conducted to investigate the impacts of active and passive immunization against synthetic inhibin and steroid-free bovine follicular fluid, respectively, on reproductive fecundity out of breeding season in Iraqi Awassi ewes. Methods: Follicular fluid was aspired from mature bovine follicles, treated with activated charcoal, and used for immunization of male rabbits for obtaining steroid free bovine follicular fluid (SFBFF) antiserum. Forty non-pregnant Awassi ewes were allocated into 4 groups (n = 10 each). At day 38 of experiment, ewes were treated with intra-vaginal MPA sponge (60 mg for 12 days). At days 0, 28, and 50, ewes were treated with 4, 2, and 2 mL of normal saline (control; C-ve), 400, 200, and $200{\mu}g$ of ovalbumin (C+ve), 400, 200 and $200{\mu}g$ of inhibin (SI group), respectively, and 4 mL of normal saline at day 0, and 4 and 2 mL of SFBFF antiserum at days 28 and 50, respectively, (AI group). After mating with Awassi rams, pregnancy and embryo number were diagnosed, at day 38 of pregnancy, using ultrasonography. Blood samples were collected at days 30, 60, 90, and 120 of pregnancy, for assessment of estradiol-$17{\beta}$ (E2) and progesterone (P4) levels. After parturition, numbers of delivered lambs were recorded. Results: The results revealed significant increase of P4 and significant decrease of E2 levels in SI and AI pregnant ewes than controls at days 30, 60, 90, and 120. Newborn number increased significantly in SI and AI treated than control ewes. Conclusion: Active or passive immunization against endogenous inhibin could augment reproductive fecundity out of breeding season in Iraqi Awassi ewes.

Investigation of useful components in soybean seeds: Purification and characterization of soybean ferritin (콩 유용성분 탐색에 관한 연구: 콩 Ferritin의 정제 및 특성)

  • Seo, Kyung-Won;Oh, Suk-Heung
    • Applied Biological Chemistry
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    • v.41 no.7
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    • pp.522-526
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    • 1998
  • Ferritin from germinated soybean seeds was purified by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, gel filtration chromatography on Sephacryl S-300, and HPLC with Bio-Scale Q2 column. SDS-PAGE analysis showed that the purified ferritin is composed of subunit with an apparent M, 21,000. The molecular mass of the native soybean ferritin estimated by gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be $510{\sim}560\;kDa$. Soybean ferritin contained 833 mol Fe/mol protein, which is 31-fold more iron than pumpkin ferritin and stained positive for iron on non-denaturing gel. Soybean ferritin cross-reacted with anti-soybean rabbit ferritin antiserum.

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Characterization of Melon necrotic spot virus Occurring on Watermelon in Korea

  • Kwak, Hae-Ryun;Kim, Jeong-Soo;Cho, Jeom-Deog;Lee, Joong-Hwan;Kim, Tae-sung;Kim, Mi-Kyeong;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.379-387
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    • 2015
  • Melon necrotic spot virus (MNSV) was recently identified on watermelon (Citrullus vulgaris) in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30-65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW) obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10-40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28-30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were $1{\mu}l/ml$ and a 1:8,000-1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb) as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.

Antigenicity of Whey Protein Hydrolysates against Rabbit Anti ${\beta}-Lactoglobulin$ Antiserum (토끼 항 ${\beta}-Lactoglobulin$ 항혈청에 대한 유청단백질 가수분해물의 항원성)

  • Lee, Soo-Won;Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Shon, Dong-Hwa;Lee, Jae-Young
    • Korean Journal of Food Science and Technology
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    • v.26 no.5
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    • pp.532-538
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    • 1994
  • In order to investigate the lowering effects of in vitro enzymatic hydrolysis by the treatment of chymotrypsin, trypsin, pancreatin, or protease from Aspergillus oryzae on the antigenicity of whey protein(WPI) against rabbit anti ${\beta}-LG$ antiserum, competitive inhibition ELISA(cELISA) and passive cutaneous anaphylaxis(PCA) test using guinea pig were performed. The results of cELISA showed that the monovalent antigenicity of the whey protein hydrolysates(WPH) to the antiserum was decreased to $10^{-1.7}{\sim}10^{-4.1}$ and less by the hydrolysis. Especially, the antigenicity of OUP(hydrolysate by protease from Asp. oryzae with preteatment of pepsin) was found almost to be removed. By the heterologous PCA the polyvalent antigenicity of the WPH was decreased to $1/2{\sim}1/128$ and less. Especially, the polyvalent antigenicity of OUN(hydrolysate by protease from Asp. oryzae without preteatments) was found almost to be removed, although OUN did not have so high degree of hydrolysis(DH) or so low monovalent antigenicity (reduced to $10^{-3.2}$). Therefore, this result was assumed to come from effective destruction of antigenic determinants on ${\beta}-LG$ in WPI, not to produce polyvalent antigenic peptides that are closely associated with induction of allergy. This finding suggested that WPH prepared by the treatment of microorganic protease from Asp. oryzae would be a material for hypoallergenic infant formula due to the removal of the polyvalent antigenicity of ${\beta}-LG$, the major milk allergen in WPI.

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Antigenicity of Whey Protein Hydrolysates Against Rabbit Anti ${\alpha}-Lactalbumin$ Antiserum (토끼 항 ${\alpha}-Lactalbumin$ 항혈청에 대한 유청단백질 가수분해물의 항원성)

  • Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Lee, Soo-Won;Lee, Jae-Young;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.436-441
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    • 1994
  • To investigate the lowering effects of in vitro enzymatic hydrolysis by the treatment of chymotrypsin, trypsin, pancreatin, or protease from Aspergillus oryzae on the antigenicity of whey protein isolate (WPI) against rabbit anti ${\alpha}-LA$ antiserum, competitive inhibition ELISA (cELISA) and passive cutaneous anaphylaxis (PCA) test using guinea pig were performed. The results of cELISA showed that the monovalent antigenicity of the whey protein hydrolysates (WPH) to the antiserum was decreased to $10^{-2.5}-10^{-5.5}$ and less by the hydrolysis. The monovalent antigenicity of the WPH hydrolyzed by trypsin, or protease from Asp. nryzae was much lowered by the pretreatment of heat denaturation. The antigenicity of the WPH hydrolyzed by chymotrypsin, trypsin, or pancreatin was much lowered by the pretreatment of pepsin. Especially, the antigenicity of TDP (trypic hydrolysate with pretreatment of heat and pepsin) was found almost to be removed. However, there was not consistency between degree of hydrolysis(DH) and the monovalent antigenicity of the WPH. By the heterologous PCA it was found that all of the PGPH lost the polyvalent antigenicity regardless of the pretreatments although WPI and ${\alpha}-LA$ had the positive high antigenicity. The results suggested that the peptides derived from ${\alpha}-LA$ in WPH could bind specific antibodies but they could not induce allergy. Therefore, it was elucidated that the allergenicity of ${\alpha}-LA$ in whey protein could be destroyed easily by the enzymatic hydrolysis.

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Cloning of Steroid $\Delta^1$-dehydrogenase Gene of Arthrobacter simplex IAM 1660

  • Bae, Moo;Bae, Song-Mee;Lee, Mi-Kyung;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.142-144
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    • 1996
  • To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.

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Characteristics of Tobacco Mosaic Virus Isolated from Wasabi (Eutrema wasabi) in Korea

  • Kim, Hyung-Moo;Lee, Kui-Jae
    • The Plant Pathology Journal
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    • v.15 no.4
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    • pp.247-250
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    • 1999
  • Wasabies showing mosaic symptoms were collected and extracted for virus purification. Tobacco mosaic virus (TMV) was identified as causal agent by electron microscopy and nucleic acid and coat protein analyses. TMV strains were determined by enzyme-linked immunosorbent assay (ELISA). TMV was identified as W and C strain in wasabi. The results of host reaction indicated that this virus induced local lesions on Nicotiana tabacum cv. Bright Yellow and N. glutinosa, leaf spots on Chenopodium amaranticolor and mosaic symptoms on wasabi. Rot shape virus particles were observed and was about 300 nm in length. About 6.5 kb single RNA molecule was observed from extracted viral RNA sample and 26 KDa coat protein was detected in denatured acrylamide gel. Infection ratio of TMV was 8% for the first cultivation year, but was 22% for the second year when TMV-W antiserum was used. The results of this experiment showed that infection ratios of both TMV-W and TMV-C strains were higher compared to that of TMV-P strain.

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The Effects of Nalbuphine on the Spontaneous locomotor activity and Primary Humoral Immune response in mice.

  • Yun, Hee-Eun;Kwak, Young-Hee;Pyo, Myoung-Yun
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.108-108
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    • 1997
  • The effects of nalbuphine.HCI on the spontaneous locomotor activity and primary humoral immune response were investigated in ICR mice. Nalbuphine was intraperitoneally administered with the dose of 130, 260, 360 mg/kg in mice. The locomotor activity such as distance traveled was observed for 90min at 10min intervals. Nalbuphine showed the biphasic dose-response relationship on the spontaneous locomotor activity. IgM plaque forming cells(PFC) in splenocytes and IgM level in antiserum were significantly decreased depending on the dose of nalbuphine when nalbuphine was administered after the immunization, but slightly increased only at the low dose in the case of nabuphine administration after the immunization(SRBC).

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