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Evaluation of Anti-cancer and Anti-proliferative Activity of Medicinal Plant Extracts (Saffron, Green Tea, Clove, Fenugreek) on Toll Like Receptors Pathway

  • Ajmal, Sidra;Shafqat, Mahwish;Ajmal, Laiba;Younas, Hooria;Tasadduq, Raazia;Mahmood, Nasir
    • Natural Product Sciences
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    • v.28 no.3
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    • pp.121-129
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    • 2022
  • Despite considerable efforts, cancer remains an aggressive killer worldwide. Chemotherapeutic drugs that are currently in use lead to destructive side effects and have not succeeded in fulfilling expectations. For centuries, medicinal plants are used for treating various diseases and are also known to have anticancer activity. The main aim of this research was to evaluate antiproliferative activity of saffron, clove, fenugreek, and green tea on Vero and MDA-MB-231 cell lines and to subsequently analyze the effect of these extracts on IRAK-4, TAK1, IKK-alpha, IKK-beta, NF-Kappa B, IRF3, IRF7 genes in Toll Like Receptors (TLRs) pathway. Antiproliferative assay was done by Neutral Red Dye uptake assay. Methanolic extract of green tea was found to be most effective against both cell lines as IC50 was achieved at least concentration of the extract. For molecular studies, MDAMB-231 cells were sensitized with methanolic extract of green tea at same IC50, and RT-PCR was performed to determine the relative expression of genes. Expression of IRAK-4, TAK1, IKK-beta, NF-Kappa B, IRF3 genes was down regulated and IRF7 and IKKalpha was upregulated. Green tea has a potential cytotoxic effect on both cell lines which was demonstrated by its effect on the expression of (TLRs) pathway genes.

Siamese Crocodile White Blood Cell Extract Inhibits Cell Proliferation and Promotes Autophagy in Multiple Cancer Cell Lines

  • Phosri, Santi;Jangpromma, Nisachon;Chang, Leng Chee;Tan, Ghee T.;Wongwiwatthananukit, Supakit;Maijaroen, Surachai;Anwised, Preeyanan;Payoungkiattikun, Wisarut;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.1007-1021
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    • 2018
  • Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.

A Growth-Stimulating Protein in Cow's Milk

  • Ki, Eun-Kyung;Park, Yong-Bok;Choi, Myung-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.4 no.3
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    • pp.59-65
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    • 1994
  • Growth-stimulating effects of cow's milk was examined using Vero cell cultre. Medium containing whole cow's milk stimulated cell growth about the same degree as that containing fetal bovine serum. The growth-stimulating factor in cow's milk was purified using hydrophobic (phenyl-sepharose) and gel filtration (Sephadex G-100) column chromatographies. It appeared that the factor is a highly hydrophobic protein, since the major growth-stimulating activity was found in the fractions eluted with 50% ethylene glycol from the phenyl-sepharose column during the purification. The purified factor showed a single band on the polyacrylamide gel electrophoresis in the presence of 1% (w/v) SDS. The factor has been found to have a relatively high molecular weight in the range of about Mr=100,000-150,000. In the presence of the purified factor (5%, w/v) in the culture medium, the incorporation of [3H]-thymidine into the cells was increased approximately 2,400-fold over that in the presence of 5% (w/v) fetal bovine serum. It seems that the growth-stimulating factor purified in this study is one of the major growth factors in the cow's milk.

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A Novel Mannose-binding Tuber Lectin from Typhonium divaricatum (L.) Decne (family Araceae) with Antiviral Activity Against HSV-II and Anti-proliferative Effect on Human Cancer Cell Lines

  • Luo, Yongting;Xu, Xiaochao;Liu, Jiwei;Li, Jian;Sun, Yisheng;Liu, Zhen;Liu, Jinzhi;Damme, Els Van;Balzarini, Jan;Bao, Jinku
    • BMB Reports
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    • v.40 no.3
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    • pp.358-367
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    • 2007
  • A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 $\mu$g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 $\pm$ 6.8), Bre-04 (41.5 $\pm$ 4.8), and Lu-04 (11.4 $\pm$ 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with $TC_50$ and $EC_50$ of 5.176 mg/ml and 3.054 $\mu$g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.

Antiherpetic Activities of Natural Quercetin Alone and in Combinations with Nucleoside Antiherpetic Agents (천연 Quercetin의 항허피스바이러스작용과 Nucleoside계 항허피스바이러스제와의 병용효과)

  • Kim, Young-So;Han, Seong-Sun
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.151-157
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    • 1999
  • In order to find less toxic antiherpetic agents, the effect of natural quercetin on the plaque formation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) was studied in vitro in cell culture monolayers employing the technique of viral plaque reduction assay. Quercetin caused a concentration-dependent reduction in the plaque formation of herpesviruses. It also exhibited more potent antiherpetic activity on HSV-1 with effective concentration $(EC_{50})$ of $18.7\;{\mu}g/ml$ than on HSV-2 with $EC_{50}$ of $24.5\;{\mu}g/ml$. The combined antiherpetic effects of quercetin with nucleoside antiherpetic agents, acyclovir and vidarabine, were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combinations of quercetin with acyclovir on HSV-1 and HSV-2 showed more potent synergism with CI values of $0.19{\sim}0.89$ for 50%, 70%, 90% effective levels than those with vidarabine with CI values of $0.43{\sim}1.46$.

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Inhibition of Herpes Simplex Viruses, Types 1 and 2, by Ginsenoside 20(S)-Rg3

  • Wright, Stephen M.;Altman, Elliot
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.101-108
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    • 2020
  • Infections by herpes simplex viruses have an immense impact on humans, ranging from self-limiting, benign illness to serious, life-threatening diseases. While nucleoside analog drugs are available, resistance has been increasing and currently no vaccine exists. Ginsenosides derived from Panax ginseng have been documented to inhibit several viruses and bolster immune defenses. This study evaluated 12 of the most relevant ginsenosides from P. ginseng for toxicities and inhibition of herpes simplex viruses types 1 and 2 in Vero cells. The effects of test compounds and virus infection were determined using a PrestoBlue cell viability assay. Time course studies were also conducted to better understand at what points the virus life cycle was affected. Non-toxic concentrations of the ginsenosides were determined and ranged from 12.5 μM to greater than 100 μM. Ginsenoside 20(S)-Rg3 demonstrated the greatest inhibitory effect and was active against both HSV-1 and HSV-2 with an IC50 of approximately 35 μM. The most dramatic inhibition-over 100% compared to controls-occurred when the virus was exposed to 20(S)-Rg3 for 4 h prior to being added to cells. 20(S)-Rg3 holds promise as a potential chemotherapeutic agent against herpes simplex viruses and, when used together with valacyclovir, may prevent increased resistance to drugs.

The Japanese Encephalitis Vaccine: Worldwide and Korean Status (일본뇌염백신: 국제적 현황과 우리나라 현황)

  • Hong, Young Jin
    • Pediatric Infection and Vaccine
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    • v.15 no.2
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    • pp.108-114
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    • 2008
  • Japanese encephalitis is the leading cause of viral encephalitis in Asia, where it accounts for up to 50,000 cases. Approximately 20% of affected patients die, and 30-50% of survivors have significant neurological sequelae. Inactivated mouse-brain derived Japanese encephalitis vaccines has been effectively implemented to control the disease effectively in Korea and several other Asian countries. However, the vaccine is expensive and difficult to produce, requires multiple doses, and has been associated with hypersensitivity reactions and rare adverse neurologicale events. The live-attenuated SA14-14-2 vaccine derived from primary hamster kidney (PHK) cells was developed in China and has been used there since 1988. Outside China, it has been licensed and used in Korea and several other Asian countries. This vaccine is effective and inexpensive. However, the lack of precedence for using a PHK cell substrate in a live-attenuated vaccine is a special issue of concern. The WHO working group has recommended additional safety studies in selected high-risk groups, as well as ongoing post-marketing studies to ensure long-term safety. Recently, a new inactivated vaccine and live-attenuated chimeric vaccine have been developed from vero cells. With this background, this article summarized the current status of Japanese encephalitis vaccination worldwide and in Korea.

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Hypoglycemic and Hypolipidemic Effects of Tectorigenin and Kaikasaponin III in the Streptozotocin-Induced Diabetic Rat and their Antioxidant Activity in vitro

  • Lee, Kyung-Tae;Sohn, Il-Cheol;Kim, Dong-Hyun;Choi, Jong-Won;Kwon, Sang-Hyuk;Park, Hee-Juhn
    • Archives of Pharmacal Research
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    • v.23 no.5
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    • pp.461-466
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    • 2000
  • Tectorigenin and kaikasaponin III from the flowers of Pueraria thunbergiana showed potent hypoglycemic and hypolipidemic effects in the streptozotocin-induced diabetic rats. Intraperitoneal administration of these two compounds with 5 and 10 mg/kg, respectively, for seven days to streptozotocin-induced rats significantly reduced the blood glucose, total cholesterol, LDL- and VLDI-cholesterol and triglyceride levels when compared with those of control group. Glycitein in which 5-OH is unlinked and tectoridin (7-O-glycoside of tectorigenin) isolated from the flowers of P. thunbergiana did not improve hyperglycemia and hyperlipidemia. In addition, tectorigenin showed in vitro antioxidant effects on 1,1-diphenyl-B-pirylhydrazyl (DPPH) radical, xanthine-xanthine oxidase superoxide anion radical, and lipid peroxidation in rat microsomes induced by enzymatic and non-enzymatic methods. We further found that tectorigenin and kaikasaponin III protected the Vero cell line(normal monkey kidney) from injury by hydrogen peroxide. From these findings, it seems likely that the antioxidant action of tectorigenin and kaikasaponin III may alleviate the streptozotocin-induced toxicity and contribute to hypoglycemic and hypolipidemic effects.

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Effects of Somatic Cell Conditioned Medium on the Chymotrypsin Resistance of Mouse Oocytes (체세포배양액이 생쥐 난자의 Chymotrypsin에 대한 내성에 미치는 영향)

  • Kim, Sung-Rye;Chung, Hye-Won;Kim, Seong-Im;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.207-216
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    • 1998
  • Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

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Cytotoxicity and Antimicrobial Effect of the Extract of Salvia plebeia (배암차즈기 추출물의 세포독성과 항균효과)

  • Shin, Min-Kyo;Kim, Seok-Keun;Lee, Sang-kon;Gang, Yeong-Seong;Kim, Seong-Su;Yang, Eun-Yeong;Lee, Hyun-Ok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.55-60
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    • 2001
  • The comparison of $IC_{50}$ values of Salvia plebeia R. Br. extracts on L1210, $P388D_1$ cancer and Vero normal cell lines showed that the n-hexane soluble extract of S. plebeia R. Br. retains the most growth-inhibitory activity against tumor cell lines. The minimum inhibitory concentration (MIC) of the extract against microorganisms were also examined. Antimicrobial activity of amocla and ketoconazole as references was compared to those of other solvent extracts such as $H_2O$, n-hexane, chloroform, ethyl acetate and methanol. The antimicrobial activity of extract had growth inhibition activity against gramnegative bacteria, gram-positive bacteria and fungi $(MIC\;>\;200\;{\mu}g/ml)$ except for n-hexane extract. Seven bacterial strains were tested for in vitro susceptibility to the extract of S. plebeia R. Br. However the n-hexane extract of S. plebeia R. Br. inhibited the growth of several bacterial strains (MIC values between 100 and $200\;{\mu}g/ml$ for gram positive bacteria, $25\;{\mu}g/ml$ for P. putida).

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