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Roles of p53 and Caspases in Induction of Apoptosis in MCF-7 Breast Cancer Cells Treated with a Methanolic Extract of Nigella Sativa Seeds

  • Alhazmi, Mohammed I.;Hasan, Tarique N.;Shafi, Gowhar;Al-Assaf, Abdullah H.;Alfawaz, Mohammed A.;Alshatwi, Ali A.
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9655-9660
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    • 2014
  • Background: Nigella Sativa (NS) is an herb from the Ranunculaceae family that exhibits numerous medicinal properties and has been used as important constituent of many complementary and alternative medicines (CAMs). The ability of NS to kill cancer cells such as PC3, HeLa and hepatoma cells is well established. However, our understanding of the mode of death caused by NS remains nebulous. The objective of this study was to gain further insight into the mode and mechanism of death caused by NS in breast cancer MCF-7 cells. Materials and Methods: Human breast cancer cells (MCF-7) were treated with a methanolic extract of NS, and a dose- and time-dependent study was performed. The $IC_{50}$ was calculated using a Cell Titer $Blue^{(R)}$ viability assay assay, and evidence for DNA fragmentation was obtained by fluorescence microscopy TUNEL assay. Gene expression was also profiled for a number of apoptosis-related genes (Caspase-3, -8, -9 and p53 genes) through qPCR. Results: The $IC_{50}$ of MCF-7 cells was $62.8{\mu}L/mL$. When MCF-7 cells were exposed to $50{\mu}L/mL$ and $100{\mu}L/mL$ NS for 24h, 48h and 72h, microscopic examination (TUNEL assay) revealed a dose- and time-dependent increase in apoptosis. Similarly, the expression of the Caspase-3, -8, -9 and p53 genes increased significantly according to the dose and time. Conclusions: NS induced apoptosis in MCF-7 cells through both the p53 and caspase pathways. NS could potentially represent an alternative source of medicine for breast cancer therapy.

Single and 28-day repeated dose toxicity studies of botulinum toxin type A in mice and rats (마우스 및 랫드에서 botulinum toxin type A의 단회 및 28일 반복투여 독성시험)

  • Jeon, Tae-Won;Kim, Ji-Young;Hyun, Sun-Hee;Kim, Nam-Hee;Lee, Sang-Kyu;Kim, Chun-Hwa;Woo, Hee-Dong;Yang, Gi-Hyeok;Jung, Hyun-Ho;Jeong, Tae-Cheon
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.57-66
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    • 2003
  • Single and 28-day repeated dose toxicity studies of botulimnn toxin type A were carried out in ICR mice and Sprague-Dawley rats, respectively. In the single dose toxicity study, botulinwn toxin was injected intraperitoneally to male and female mice at a single dose of 40, 59, 89 133 and 200 ng/10 ml saline/kg. All animals died from 59 ng/kg group. Some clinical signs, such as decrease in locomotor activity, dyspnea, prone position and ptosis, were observed in most of both sexes from 59 ng/kg group, but no signs were seen in all animals at 40 ng/kg group. The results showed that the median lethal dose of botulinum toxin might be in the range of 40-59 ng/kg in both sexes. In the repeated dose toxicity study, the test material was administered intradermally for 28 days at doses of 0 (vehicle-treated control), 1.25, 2.5, 5.0 and $10.0ng/head/50{\mu}{\ell}$ saline in male and female rats. No test material-related changes were noted in survivals, clinical signs, food and water consumptions and gross finding in any group. Botulinum toxin treatment significantly decreased the body weight gain rate in male of 5.0 ng/head group and over and in female of 10.0 ng/head group compared to vehicle-treated control. One or more relative organ weights (i.e., spleen, thymus, liver and kidney) were increased significantly from 5.0 ng/head group compared to vehicle-treated control in both sexes. Serum biochemistry revealed increases in aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine phosphokinase, total protein and albumin in male, and increases in AST and ALT and decreases in $K^+$ and $Cl^-$ in female without dose-pendent manners. In the histopathological study, physical stimulation by needle caused slight inflammations of dennis. In addition, botulinum toxin treatment induced denervation of nerve cell and disuse of muscle, resulting in atrophy of skeletal muscle in both sexes from 2.5 ng/head group. When the antibodies to toxin were determined in all animals, a significant increase in serum antibodies was observed from 5.0 ng/head group. The results showed that the NOAEL of botulinum toxin might be 1.25 ng/head for 28-day repeated dose toxicity in rats.

Inhibitory effect of by Ojeok-san lipid accumulation in high fat diet-induced obesity mice and 3T3-L1 adipocytes (오적산 (五積散)의 고지방식이 마우스 지방축적억제 및 3T3-L1지방세포에서의 비만 조절 기전에 관한 연구)

  • Choi, Hye-Min;Moon, Sung-Ok;Lee, Hee-Hyun;이, 화동
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.121-128
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    • 2015
  • Objectives : Ojeok-san (OJS), an oriental herbal formula, has been used in Asian countries including Korea, China and Japan to treat the common cold and illnesses including fatigue and gastrointestinal disorders. The purpose of this study was to examine the anti-obesity effect and molecular mechanism of OJS, on adipogenesis in 3T3-L1 cells. Also, the effects of OJS in obese mice fed a high-fat diet on adiposity were examined.Methods : Preferentially, we analyzed the component of OJS and measured the stability of its component in OJS according to study periods using high performance liquid chromatography (HPLC). In vitro, 3T3-L1 cells were treated with OJS (50 to 200 μg/mL) during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining. The expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. For anti-obesty effect in vivo, we experimented for 8 weeks with four group (normal diet (CON), high-fat diet (HF), high-fat diet with OJS (HF+OJS) and high-fat diet with Bang-pung-tong-sung-san (HF+BTS) in comparison group HF+OJS).Results : OJS showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, OJS significantly reduced the expression levels of several adipocyte marker genes including proliferator activated receptor-γ(PPAR-γ) and CCAAT/enhancer-binding protein-α(C/EBP-α). Also OJS-administered mice showed significant inhibitory of body weights and abdominal adipose tissue weights.Conclusions : This study showed that traditional medicine OJS has an anti-obesity effect in vitro and in vivo. Thus, OJS could be developed as a supplement for reduction of body weight gain induced by an obesity.

EFFECT OF METHANOL EXTRACT OF CNIDII RHIZOMA ON THE FUNCTION OF RECEPTORS FOR GABA AND GLYCINE (천궁(Cnidii Rhizoma)의 메탄올 추출물이 GABA 및 Glycine 수용체에 미치는 영향)

  • Lee, Jong-Tae;Lee, Keung-Ho;Park, Jae-Hong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.55-66
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    • 2005
  • Cnidii Rhizoma (CR) was subjected to extraction with 70% methanol and tested to determine whether it has anxiolytic activity in mouse by employing staircase and rotarod tests. In addition, to understand the mechanism of anxiolytic action, CR, picrotoxin, yohimbine, isoniazid and strychnine were utilized to deliniate the potential involvement of GABA and glycine receptors in the action of Cnidii Rhizoma. To gain insights into the safety of Cnidii Rhizoma extract, behavioral and MTT tests were carried out. The results were obtained as follows: 1. CR extract had little effect on climbing numbers in the stair case test. 2. CR extract had considerable anti-anxiety effects as evidenced by the reduction of rearing numbers in the stair case test. 3. CR extract had little effect on muscle relaxation. 4. Anxiolytic actions of CR extract appeared to be mediated by glycine receptor activation. 5. Cytotoxicity in the neuronal cell was not observed and no strange behaviors were found. In short, these results indicate that CR extract has the ability to exert anxiolytic activity, possibly by activating glycine receptor with little side effects in mouse.

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Effect of Various Fat and Oil Types on Weight Gain and Liver Metabolism in Fasting or Normal Diet Rats (불포화도가 다른 유지가 단식과 정상식이 흰쥐의 체중 및 간대사에 미치는 영향)

  • 서화중;박현희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.1
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    • pp.19-26
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    • 1993
  • In this study rats in fasting or fed protein free restricted diet including only fat showed much lowered level of serum cholesterol and triglyceride accompanied by utmost weight loss and high level of blood urea nitrogen indicated the tissue degradation, especially in liver with signs of damage or necrosis of hepatic parenchymal cell leading to elevated glutamic pyruvate transaminase value and to death. Rats fed only perilla oil in starvation or as fat source in normal diet dropped down the level of serum cholesterol and triglyceride compared to beef tallow fed rat. But with evidence of glutamic pyruvate transaminase values which was significantly elevated long term ingestion of perilla oil is likely to cause the lesion or any damage of hepatic function.

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The Microstructure of Magnetite Coated on Honeycomb and Characteristics of CO2 Decomposition (허니컴에 코팅한 마그네타이트의 미세구조 및 CO2 분해특성)

  • 윤용운;김은배;이병하;고태경;오재희
    • Journal of the Korean Ceramic Society
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    • v.41 no.5
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    • pp.410-416
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    • 2004
  • In this study, we fabricated magnetite coated on a cordierite honeycomb which has complex shape by ultrasound-enhanced ferrite plating. The effects of the plating condition on the formation of the magnetite and its microstructure were investigated. The magnetite coated on the honeycomb became an oxygen-deficient ferrite by H$_2$ gas reduction, then the effects of the molar concentrations of ammonium acetate for $CO_2$ gas decomposition have been studied. As the molar concentration of a pH buffer($CH_3$COONH$_4$, 0.1946∼0.3892 M) solution increased, the average particle size increased about 200∼250 nm. The magnetite coated on the honeycomb was reduced by H$_2$ gas for 2 h at 30$0^{\circ}C$. The inner pressure change in the cell began to occur at 315∼34$0^{\circ}C$. The H$_2$-Reduced magnetite coated on the honeycomb at 35$0^{\circ}C$ contained an oxygen deficient magnetite and $\alpha$-Fe phase. The thermogravimetric analysis with H$_2$ reduction and $CO_2$ decomposition were carried out with the magnetite coated on the honeycomb. A weight loss in process of H$_2$ reduction occurred between 32$0^{\circ}C$ and 34$0^{\circ}C$, while a weight gain was observed during the $CO_2$ decomposition.

Characterization of Pv92, a Novel Merozoite Surface Protein of Plasmodium vivax

  • Lee, Seong-Kyun;Wang, Bo;Han, Jin-Hee;Nyunt, Myat Htut;Muh, Fauzi;Chootong, Patchanee;Ha, Kwon-Soo;Park, Won Sun;Hong, Seok-Ho;Park, Jeong-Hyun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.385-391
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    • 2016
  • The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.

A Study of Dewatering Phenomena of Potato Slice Cytorrhysed by High Molecules (고분자 용액의 세포 압착 현상에 의한 감자 절편의 탈수 현상에 관한 연구)

  • Choi, Dong-Won;Shin, Hae-Hun;Kim, Jong-Geu
    • The Korean Journal of Food And Nutrition
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    • v.19 no.4
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    • pp.358-365
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    • 2006
  • To study simultaneous water and solute transport kinetics during soaking in concentrated solution, the influence of the concentration and molecular weight of the solute(polyethylene glycol(PEG) and NaCl) in the soaking solution and the temperature on the water loss and solute gain rates were observed by using a model vegetable tissue(potato). When potato slices$(4cm{\times}4cm{\times}0.1cm)$ soaked in 60% PEG solutions, the water loss rate of the early phase decreased with increasing of the molecular weight of PEG from 200 to 6,000, while the final water loss increased with increasing the molecular weight of PEG and it reached to 80%. The cell wall of potato tissue was permeable to NaCl and PEGs of which average molecular weight is smaller than 400 but it was not permeable to PEG 600 and larger molecules. PEG which has average molecular weight below 600 induced plasmolysis and those above 600 induced cytorrhysis. The water loss rate of potato sample soaked in smaller molecular weight PEG solution was faster than those soaked in higher molecular weight PEG solution before cytorrhysis happened. The water loss rate was reversed after cytorrhysis happened. The volume change of potato within the first 60 minutes was larger in low molecular PEG solution but the final ratio of decreasing volume was larger in high molecular PEG solutions. In PEG 200 solution, the potato tissue was slightly shrinked without shape change. However, in PEG 4,000 solution, volume of potato was reduced significantly and potato tissue was twisted.

Hypoglycemic Effect of the Giant Embryonic Rice Supplementation on Streptozotocin-induced Diabetic Rats (거대배아미 식이에 의한 Streptozotocin 유발 당뇨 흰쥐의 혈당강하 효과)

  • Lee, Youn-Ri;Nam, Seok-Hyun;Kang, Mi-Young
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.427-431
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    • 2006
  • This study was performed to investigate the influence of giant embryo feeding on hypoglycemia in streptozotocin-induced diabetic rats. Streptozotocin -induced diabetic rats were fed four kinds of experimental diets such as corn starch diet as a control, polished rice diet, brown rice diet and giant embeyonic rice diet for 6 weeks. The body weight gain diabetic of control was signaficantly suppressed compared to that of the normal. The supplementation of giant embryo increased the surivival rate. Feeding of giant embryo appeared to have ameliorating effects on diabetic symptoms including features such as polyphasia, polyuria. Giant embryo shortened gastrointestinal transit time and increased total fecal weight, total fecal dry weight and fecal water content compared with diabetic control. Giant embryo showed fasting blood glucose lowering effect compared with diabetic control. The disaccharidase activities in proximal part of intestine such as maltase sucrase and lactase in giant embryo feeding groups were lower than diabetic control. The results of this study show that giant embryo supplementation may have a beneficial Veffect on the hypoglycemia may be useful in the diet therapy for diabetic.

System Performance Improvement of IEEE 802.15.3a By Using Time Slot Synchronization In MAC Layer (UWB MAC의 Time Slot 동기를 통한 시스템 성능 개선)

  • Oh Dae-Gun;Chong Jong-Wha
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.43 no.3 s.345
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    • pp.84-94
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    • 2006
  • In this paper, we propose the algorithm to reduce guard time of UWB MAC time slot for throughput gain. In the proposed draft by multiband ofdm alliance (MBOA), Guard time of each medium access slot (MAS) is composed of shortest inter-frame space (SIFS) and MaxDrift which is the time caused by maximum frequency offset among devices. In this paper, to reduceguard time means that we nearly eliminate MaxDrift term from guard time. Each device of a piconet computes relative frequency offset from the device initiating piconet using periodically consecutive transferred beacon frames. Each device add or subtract the calculated relative frequency offset to the estimated each MAS starting point in order to synchronize with calculated MAS starting point of the device initiating piconet. According to verification of simulations, if the frequency offset estimator is implemented with 8 decimal bit, the ratio of the wasted time to Superframe is always less than 0.0001.