• Journal of the korean society of oceanography
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• v.34 no.3
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• pp.167-171
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• 1999

Since toxic Alexandrium catenella and non-toxic A. fraterculus are morphologically similar, they are difficult to discriminate under the light microscope. However, a novel technology, such as fluorescein isothiocyanate (FITC)-conjugated lectin probes enables easy and rapid differentiation. Toxic A. catenella bound seven different lectins, whereas the non-toxic A. fratercuzus did not bind Arachis hypogaea (PNA) lectin. In addition, Pseudo-nitrschia species in this study were also difficult to identify to species level with light microscope techniques, but it was possible to classify them using fluorescent lectins. Pseudo-nitzschia multistriata, P. subfraudulenta and P. pungens bound Canavalia ensiformis (ConA), whereas P. subpaclfica did not, and P. pungens also bound Ricinus communis (RCA). These results imply that lectin could be used as a critical tool in the differentiation of P. multistriata, P. subfraudulenta and P. pungens. However, P. subpacifica was not differentiated by the lectins tested. Therefore, it isconcluded that lectin probes are useful for discriminating toxic A. catenella from non-toxic A. fraterculus, and for the identification of some Pseudo-nitzschia species. In addition, this method has a great potential to speed and detection between non-toxic and toxic harmful algal blooms (HABs) in Korean biotoxin monitoring systems.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.381-381
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• 2000

Lectin binding assay was conducted on 3 A. tamarense isolates (AT-A, AT-2 and AT-6). Fatty acid composition of all 3 isolates was analyzed, and total carotenoid content and $\beta$-carotene were also determined. AT-A and AT-2 treated with different lectins in this study showed the positive response, whereas potentially toxic AT-6 did not bind DBA lectin, regardless of different growth phase, but conjugated ConA, PNA, RCA, SBA, UEA and WGA. It is possible that DBA is a desirable method for rapid and easy discrimination of highly toxic A. tamarense. AT-A, AT-2 and AT-6 comprised saturated fatty acids (49.0-61.9%), monounsaturated fatty acids (8.0-20.5%) and polyunsaturated fatty acids (23.2-30.5%). In particular, 22:6 (n-3) polyunsaturated fatty acid in AT-6 had a high abundance, compared with AT-A and AT-2. However, carotenoid content and $\beta$-carotene were not contributed to discriminate each isolate. Due to variability in biochemical composition at different isolates, possibly DBA and 22:6 (n-3) polyunsaturate fatty acid provide a good information for discrimination of AT-6.

• Journal of Life Science
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• v.14 no.2
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• pp.301-308
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• 2004

The morphology of the apical pore complex, the first apical plate and the posterior sulcal plates in a new isolate of Alexandrium tamarense (Lebour) Balech from the Bay of Chinhae was compared with other that of toxic strains of A. tamarense previously isolated from Korean waters. Although this isolate was morphologically identical to these toxic strains, high performance liquid chromatography and mouse bioassay showed no evidence of toxin production. The nontoxic A. tamarense strain showed a strong positive binding activity with PNA lectin, indicating a high density of lactose and galactose residues on the cell surface, and in SDS-PAGE and Western blot analysis a unique protein of about 21-kDa molecular sizes was observed. These findings demonstrate that the use of PNA and immunobioassay could be used to discriminate between toxic and nontoxic strains of A. tamarense.

• Toxicological Research
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• v.11 no.2
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• pp.193-197
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• 1995

Antibody-toxin conjugates, termed immunotoxins, are currently being evaluated as potential new anticancer agents and one of the most extensively studied toxins for construction of immunotoxin is ricin which exists in the seeds of castor bean, Ricinus communis. Another toxic lectin from castor bean is RCA (Ricinus communis agglutinin). Both toxins are very homologous. We reported the puriffcation procedure and biological properties of ricin from the Korean castor bean in another place and here we report those of RCA. The purified RCA shows three bands on denatured SDS PAGE while ricin shows two bands. On cultured $K_{562}$ cells ricin and RCA both inhibit the multiplication of cells extensively. $30{\mu}g/ml$ of ricin shows 73% of inhibition rate at day 4 compared to 68% in same condition of RCA. The inhibition of multiplication of cells are directly proportional to the concentration of toxins and the incubation period. In every case ricin was more toxic than RCA. The $LD_{50}$ dose of ricin on ICR mice was 60 ng at day 3 but that of RCA was $10{\mu}g$.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.250-254
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• 1998

Harmful micro algae isolated from Korean coastal waters, were tested with FITC-conjugated lectins and observed by epifluorescent microscopy to distinguish each other. Strain-specific sugar composition at the cell surface was suggested by the affinity of lectins to different microalgae. The micro algae Cochlodinium polykrikoides (CP-1) and Gymnodinium $A_3\;(GA_{3-1}\;1)$, are morphologically similar, but exhibited different binding activity with the lectins ECA, HPA and WGA. In Peridiniales, the micro alga Alexandrium tamarense (AT) bound HPA and WGA, but Scrippsiella trochoidea (ST-1) did not bind those lectins. Three species of Prorocentrum also exhibited different binding specificity with HPA, PHA and SBA. A non­toxic Korean isolate of Heterosigma akashiwo (HA-2) bound ConA, PEA and UEA. These results suggest that lectins are useful in discriminating morphologically similar species, as well as different species or strains within the same genus.

• Korean Journal of Pharmacognosy
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• v.42 no.3
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• pp.246-252
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• 2011

Korean mistletoe Lectin (KML-C) is composed of A and B sub-chain. B chain binds to carbohydrates on cell surface and A chain hinders translation and induces an apoptosis as a RIP (ribosome inactivating protein). KML-C has very strong biological activities, it has seriously limits to use as a cancer therapy or adjuvant because of its toxicity to normal cells. This study is therefore conducted to see if B chain of KML-C might have immunological activity, especially adjuvant activities with less toxicity. We isolated B chain from KML-C using the lactose affinity chromatography, and examined their immunoadjuvant activity. The isolated B-chain did not show any cytotoxicity against tumor cell, RAW264.7, and P388D1 while KML-C had a very strong toxicity. This non-toxic effect was observed also by in-vivo study. Both humoral and cellular immunities were observed ; the antibody titer was increased when the mice were immunized with B-chain used as adjuvant like Freund's adjuvant, indicating that B chain of mistletoe lectin alone might be used for adjuvant; it also increased DTH in cellular immunity. These results suggest that B-chain of KML-C might be used for adjuvant used for the production of antibody or vaccine with less toxicity.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.225-234
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• 2006

In the present study toxicity and immunostimulating activity of the lectin(KML-C), which was extracted from Korean mistletoe(Viscum album coloratum) were investigated in swine. To determine the toxicity, lectin was injected into thigh or cervical muscles of 4-week-old piglets(Landrace) and observed clinically and pathologically. For determination of the immnunostimulating activity, lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine of Aujeszky's disease virus(ADV)(NYJ1-87) which was inactivated by 0.2% formalin was injected into the cervical muscle of antibody-negative piglets in the same age group. Subpopulation of the immune cells and serum neutralizing(SN) antibodies in the piglets were examined after vaccination, and resistance of the piglets against challenge by virulent NYJ1-87 was further examined. The results were also compared with those from piglets injected with aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccine of inactivated NYJ1-87 and NYJ1-87 vaccine without adjuvant, and the results are as follows. By injection of lectin with $30{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets died after signs such as dyspnea, fever, systemic erythema and subcutaneous hemorrhages, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with $7{\mu}g/kg$ of body weight to the thigh muscle, all of 12 piglets showed signs such as edema and cutaneous hemorrhage in the injected area, lameness and depression, and lesions pertaining to poisonous hepatitis and dysfunction of kidney were observed. By injection of lectin with 1, 3 and $5{\mu}g/kg$ of body weight to the thigh muscle of each one piglet, signs such as congestion, induration and grayish coloration in the injected area, depression and inappetence were observed in all piglets. Toxic changes were also observed in the liver and kidney of piglets by lectin of 3 and $5{\mu}g$. By injection of lectin with 0.5 and $0.7{\mu}g/kg$ of body weight to the cervical muscle of each 9 piglets, all piglets were clinically normal and there were no significant changes in blood counts and chemistry values. Whereas, epithelial swelling and vacuolation of convoluted tubules were observed from one piglet injected with lectin of $0.7{\mu}g$, and necrosis and fibrosis of muscular fiber were observed in the muscle of one piglet injected with lectin of $0.5{\mu}g$. Only population of sIgM+ B lymphocytes increased among immune cells in all of 15 piglets immunized with lectin($0.7{\mu}g/kg$ of body weight)-adjuvanted vaccine, while compared to those in $Al(OH)_3$-adjuvanted vaccine and vaccine without adjuvant. No additional stimulation to the immune cells was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. In piglets immunized with lectin-adjuvanted vaccine, SN titers in reciprocal values for loge were 1.3-4.0 at 1-4 weeks after vaccination, which was similar to those with 1.0-3.3 by vaccine without adjuvant but lower than those with 2.0-5.7 by $Al(OH)_3$-adjuvanted vaccine. Also, no additional increase in the SN titers was recognized when lectin was added to $Al(OH)_3$-adjuvanted vaccine. Piglets immunized with lectin-adjuvanted vaccine were resistant to challenge by the virulent NYJ1-87 at 4 weeks after vaccination, and the SN titers reached to 5.0 one week after challenge, which was higher than those with 4.0 by vaccine without adjuvant but somewhat lower than those with 7.7 by $Al(OH)_3$-adjuvanted vaccine.

• BMB Reports
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• v.36 no.2
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

• Korean Journal of Pharmacognosy
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• v.24 no.2
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• pp.177-182
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• 1993

The pericarp of Cornus officinalis is well known famous medicinal drug in oriental countries. In this work, we have tried to evaluate the toxicity and also to find the lectin components from this seed. The lyophilized seed extract was lethal to experimental mouse at $250{\sim}300mg/kg$ and this toxic components were related to proteins. The lectins components were partially purified from the extract by ion exchange column chromatography. These lectins were relatively stable at temperature variations and also stable at pH $4{\sim}7$. The activity of these lectins did not inhibit by common carbohydrates molecules.

• YAKHAK HOEJI
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• v.35 no.5
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• pp.444-455
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• 1991

Two kinds of new lectin fractions (LOA-I, LOA-II) were obtained from loach (Misgurnus spp.) meat by 0.15 M NaCl extraction, salt fractionation, ion exchange and hydroxyapatite column chromatographies. On polyacrylamide gel electrophoresis, LOA-I exhibited one major and a few minor bands, but LOA-II exhibited three minor bands. The partially purified loach lectins agglutinated not only erythrocytes of human B and AB type, rabbit, dog, but also murine splenic lymphocytes. Agglutinability was relatively labile at various pH and stable at increasing temperature, but was not affected by tested several metal ions. By the sugar specificity test, D-glucosamine and metyl-$\beta$-galactopyranose inhibited agglutinating activity at a final concentration of 3 mM. The lectins contained relatively high amounts of aspartic acid, valine and leucine, but sulfur containing amino acids, cystein, methionine and isoleucine were not determined. LOA-I, LOA-II lectins were nonmitogenic toward murine lymphocytes.