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.
It is well-known that Korean mistletoe (Viscum album) extract has an immune activity and anticancer effect. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to examine whether this extract might activate human peripheral monocyte to produce tumor necrosis $factor-\alpha$$(TNF-\alpha)$. To examine the effect of M11C on the production of $TNF-\alpha$ from monocyte, the monocyte were stimulated by the M11C, and then collected the supernatant (M11C stimulated monocyte-conditioned media; MCM). MCM was treated to the $TNF-\alpha$ sensitive L929 cells, and then L929 cytotoxicity was measured by means of MTT. MCM had cytotoxic effect on L929. And the cytotoxic effect of MCM on L929 was almost abolished by $anti-TNF-\alpha$ antibody. These data indicated that MCM contained $TNF-\alpha$, suggesting the $TNF-\alpha$ generation from M11C-stimulated monocyte. This suggestion was confirmed from the data that $TNF-\alpha$ was highly detected in MCM by immunoblotting technique. M11C effect on $TNF-\alpha$ production from monocyte was in the dose and stimulating time dependent manners. Also the effect of M11C on the expression of $TNF-\alpha$ mRNA from monocyte was shown in the dose and stimulating time dependent manners. As a result, Korean mistletoe extract, M11C, could be used for an immunostimulator.
This study was conducted to study the seasonal changes of relative water relations parameters by pressure-volume curves of Korean mistletoe (Viscum album var. coloratum) leaves for understanding ecological characteristics. In growing season, the osmotic potentials at full turgor (${\Psi}_o^{sat}$) and at incipient plasmolysis (${\Psi}_o^{tlp}$) decreased, while increased the maximum bulk elastic modulus of the cell wall ($E_{max}$) and relavive water content ($RWC^{tlp}$). Korean mistletoe in Quercus variabilis and Korean mistletoe in Quercus mongolica in November showed best maximum perssure potential (${\Psi}_{P,\;max}$). Pressure potential (${\Psi}_P$) and water potential (${\Psi}_L$) in Korean mistletoe in Quercus variabilis were rapidly decreased with decreasing of relative water content. The values of $RWC^{tlp}$ in November were all above 84% showing that the function of osmoregulation is somewhat better, and symplastic water content (Vo/DW) and maximum water content (Vt/DW) were variable seasonally. Thus, responses to water relations of Korean mistletoe in Quercus variabilis and Korean mistletoe in Quercus mongolica such as ${\Psi}_o^{sat},\;{\Psi}_o^{tlp},\;E_{max},\;{\Psi}_{P,\;max},\;RWC^{tlp}$ showed increaing drought tolerance with increasing of leaf aging.
Effects of Korean mistletoe extracts (KM-110), Viscum album coloratum on the specific and non-specific immune responses of Japanese eel Anguilla japonica were examined. The optimal concentration not showing toxicity of KM-110 was determined to $30-40{\mu}g/ml$ in vitro and $100{\mu}g$/100 g of fish in vivo. Even $1000{\mu}g$ of KM-110/100 g of fish did not show any clinical problem in fish though the levels of toxic parameters were slightly increased. In terms of antibody production, KM-110 significantly elicited more antibody production than FCA or $\beta$-glucan. $\beta$-glucan plus KM-110 group synergistically enhanced antibody production. There was no significant difference between KM-110 and KM-110 plus $\beta$-glucan group. The ROI production by head kidney (HK) leucocytes of eel injected with 500 or $1000{\mu}g$ KM-110 was significantly (P<0.05) enhanced than the control and FCA-treated group. Maximum increase in the NBT reduction value was observed in $1000{\mu}g$ KM-110 group but no significant difference was found between 500 and $1000{\mu}g$ KM group. The level of serum lysozyme activity was significantly (P<0.05) higher in the 500 and $1000{\mu}g$ KM-110- or FCA-treated group than in the control and $200{\mu}g$ KM-110 group. The phagocytic activities of HK leucocytes isolated from eel injected with 500 and $1000{\mu}g$ KM-110 were significantly (P<0.05) higher than $200{\mu}g$ KM-110 and PBS-injected control group. Korean mistletoe appeared to be a good activator of the specific and non-specific immune responses of Japanese eel.
Park, Jong-Heum;Ji, Seung-Taek;Hyun, Chang-Kee;Chin, Koo-Bok;Shin, Heuyn-Kil
Korean Journal of Food Science and Technology
/
v.32
no.2
/
pp.461-468
/
2000
To investigate the antigenotoxicity of Korean mistletoe using Comet assay, the crude extract was divided into 4 fractions, i.e. fraction I (MW range over 14,000), fraction II $(8,000{\sim}14,000)$, fraction III $(3,500{\sim}8,000)$, and fraction IV (below 3,500) by molecular weight fractionation. In the non-tumoral 3T3 cells, fraction IV could reduce DNA damage induced by MNNG in a dose dependent pattern while fraction I and III which were known to contain lectins and viscotoxins, respectively, did not show the activity. By heat treatment, the antigenotoxic activity of faction IV, though was gradually diminished according to heating time, was found to be maintained significantly. From the Sephadex G-25 gel filtration chromatography, a more purified fraction responsible for the activity of faction IV was obtained from the latter part of total elute. Therefore, it was concluded that the antigenotoxic components of Korean mistletoe were water soluble substances of MW below 1,000 and there is a possibility of utilization as a material of functional foods for chemoprevention.
Park, Jong-Heum;Kim, Su-Min;Sung, Nak-Yun;Song, Du-Sup;Byun, Eui-Baek;Kim, Jae-Kyung;Song, Beom-Seok;Lee, Ju-Woon;Kim, Jae-Hun
Journal of Radiation Industry
/
v.7
no.2_3
/
pp.183-190
/
2013
Mistletoe (Viscum album) has been widely used as a functional food material for various therapeutic purposes from ancient time. In this study, we examined anti-diabetic and cytotoxic activities of heated-treated mistletoe and the effects of gamma-irradiation on its activities. Heat-treated mistletoe extract was prepared by heating during different time (3, 6, 9 and 12 h) and gamma-irradiated with different doses of 0, 10, 30, 50, 70 and 100 kGy. Heat-treated mistletoe extracts showed a concentration-dependent cytotoxicity on rat insulinoma RINm5F cells and the effect was gradually decreased as heating time increased up to 12 h. 12 h heat-treated extract was no cytotoxic. Gamma-irradiation enhanced the reduction of heat-treated mistletoe-induced cytotoxicity and the decreasing effect was an irradiating dose-dependent. In particular, all of 70 kGy irradiated and heat-treated mistletoe extracts did not showed the cytotoxicity and the effect was comparable to 12 h heat-treated mistletoe extract. Among those extracts, 3 h heat-treated mistletoe extract gradually increased the insulin secreting activity by gamma-irradiation and the effect was the best at 70 kGy, whereas 12 heat-treated extract was no effect. On the test of ${\alpha}$-glucosidase inhibitory activity, 3 h heat-treated mistletoe extract showed the concentration dependent effects and gamma-irradiation induced more activity at 70 kGy, compared to non-irradiated 3 h and 12 h heated mistletoe extracts. These results suggest that the combination of heat treatment and gamma-irradiation might be more effective than only heat-treatment for improving the anti-diabetic activity of mistletoe extract and reducing its cytotoxicity.
Global warming is a key challenge subjecting animals to heat stress conditions resulting in multiple physiological alterations in tropical climate. Dietary approach seems to be the more friendly approach to curb the adverse effects of heat stress in rabbits. Some herbs have been categorized to have high potential for promotion of immune responses for amelioration of heat stress. Thus, this research aims to evaluate the potential of Mistletoe (Viscum album), Moringa (Moringa oleifera) and Phyllanthus (Phyllanthus amarus) leaf meal as herbal supplements for the alleviation of heat stress in female rabbits by measuring improvement in sex and stress hormonal responses in serum biochemistry. 80 Rabbit does were exposed to 4 dietary groups supplemented with each of Mistletoe, Moringa, Phyllanthus and a control in an 84-day trial at the summit of thermal stress in South west Nigeria. Growth indices were monitored throughout the study, blood samples were compiled at the end of the trial to assess serum biochemistry, stress and sex hormonal responses of the Does using standard protocols. The results revealed that final weight and weight gain of Does fed on Phyllanthus were significantly (p < 0.05) higher (11.46% and 14.25%, respectively) than Does on control. The herbal supplements enhance glucose, protein, albumin and globulin, reduced cholesterol, and creatinine of Does under heat stress conditions. Among the herbal treatment groups, mistletoe, moringa and phyllanthus had 12.42%, 18.39% and 16.90%, respectively, lower corticosterone than control groups which had 39.76ng/ml. Triiodothyronine of Does fed control were significantly (p < 0.05) lower than Does on Moringa oleifera and Phyllanthus amarus supplements. Estradiol and Follicle stimulating hormone of rabbit Does fed on moringa supplement were significantly (p < 0.05) higher than other treatments. In conclusion, the herbal supplements tend to mitigate the detrimental outcome of thermal stress on Does by suppressing stress hormones. Moringa oleifera and Phyllanthus amarus enhanced sex hormones while Phyllanthus amarus confered growth promoting effects on the Does.
The lectin was purified through 0.15 M NaCl extraction, ammonium sulfate precipitation, sepharose 4B affinity chromatography and gel filtration using sephadex G-150 from the leaves of Visum album collected in Mt. Duk Yu. The final gel filtration step resulted in 11.64 folds purification with 0.14% of recovery yield. We also performed biochemical characterization of the purified Visum album lectin. HPLC analysis of lectin purified by gel filtration revealed a singel peak. The analysis of the purified lectin by SDS-PAGE showed a tetramer composed of two identical subunits with molecular weights of 32 and 30 kDa. The lectin was a glycoprotein containing 14.4% carbohydrate, which consist of glucose, fructose, arabinose and xylose, and the amino acids such as phenylalanine, lysine and tyrosine. The purified lectin agglutinated human red blood cell types with similar potency, but when tested against red blood cells from mouse, bovine, rabbit, chicken and porcine, significant difference in potency were observed. Hemaggluting activity was inhibited by D-galactose, D-mannose, D-lactose and D-raffinose, but not by D-glucose, D-glucosamine, D-mannosamine, L-fructose, D-xylose, D-arabinose, D-galacturonic acid, D-fructose, L-rhamnose and N-acetyl-D-galactosamine. The optimal pH and thermal stability of the purified lectin were pH 4.0-7.0 and 20-5$0^{\circ}C$, respectively.
The objective of this study was to clarify the distribution, host preference and habitat characterstics of the endangered mistletoe species, Loranthus tanakae in Korea. Seven sites were selected for sampling in Gangwon Province, located in middle of Korea. Parameters including DBH, attached height, twig death, and the number of L. tanakae and Viscum album var. coloratum per tree for all infected individuals were investigated. One thousand, two hundred and seventy L. tanakae plants were found on the 420 host trees with 665 V. album var. coloratum on 227 of the trees. The infested host trees belonged to eight species and one variety species from six genera and five families. The highest percentage (86.9%) of individuals of L. tanakae were growing on Fagaceae. The infected trees occurred between altitudes of 420 m and 1,250 m above the sea level, but most of them were between 600 m and 900 m. The mean DBH and heights of parasitism were 38.6 cm and 13.0 m, respectively. Twig death was also observed on 300 host trees (71.4%) and 1.5 twig per tree. Most of the populations and the distribution of L. tanakae were highly affected by DBH out of some individual tree characteristics.
A triterpene was isolated as a cytotoxic principle from the dichloromethane extract of Korean mistletoe (KM; Viscum album colora turn) by repeated silica gel chromatography and recrystalli-zation. In in vitro analysis of cytotoxic activity using various human and murine tumor cell lines, the dichloromethane extract of KM was highly cytotoxic against these cells. We isolated the most active compound, referred to VD-3, from the dichloromethane extract of KM. The VD-3 was shown to be less cytotoxic to normal cells (murine splenocytes). From the identification of the chemical structure of VD-3 by spectral data and chemical synthesis, the compound was proven to be epi-oleanolic acid. Tumor cells treated with VD-3 showed a typical pattern of apo-ptotic cell death, such as apparent morphological changes and DNA fragmentation. These results indicate that epi-oleanolic acid is an important compound responsible for antitumor activity of KM.
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