• Title/Summary/Keyword: in vitro effect

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A Tuber Lectin from Arisaema jacquemontii Blume with Anti-insect and Anti-proliferative Properties

  • Kaur, Manpreet;Singh, Kuljinder;Rup, Pushpinder Jai;Kamboj, Sukhdev Singh;Saxena, Ajit Kumar;Sharma, Madhunika;Bhagat, Madhulika;Sood, Sarvesh Kumar;Singh, Jatinder
    • BMB Reports
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    • v.39 no.4
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    • pp.432-440
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    • 2006
  • A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

Protein Tyrosine Kinases, $p56^{lck}\;and\;p59^{fyn}$, MAP Kinase JNK1 Provide an Early Signal Required for Upregulation of Fas Ligand Expression in Aburatubolactam C-Induced Apoptosis of Human Jurkat T Cells

  • BAE MYUNG AE;JUN DO YOUN;KIM KYUNG MIN;KIM SANG KOOK;CHUN JANG SOO;TAUB DENNIS;PARK WAN;MOON BYUNG-JO;KIM YOUNG HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.756-766
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    • 2005
  • The signaling mechanism underlying aburatubolactam C-induced FasL upregulation was investigated in human Jurkat T cells. After treatment with aburatubolactam C, the src-family PTKs $p56^{lck}\;and\;p59^{fyn}$, and MAP kinases ERK2 and JNK1, were activated prior to FasL upregulation; Both $p56^{lck}\;and\;p59^{fyn}$ were directly activated 2.4- and 2.2-fold, respectively, in vitro by aburatubolactam C. The aburatubolactam C-induced cellular changes, including the activation of ERK2 and INK1, and FasL upregulation, were completely prevented by the PTK inhibitor genistein. The activation of protein kinase C (PKC$\delta,\;\epsilon\;and\;\mu$ was also induced following aburatubolactam C treatment. Although the activation of $p56^{lck}$ and tyrosine phosphorylation of the cellular proteins were not blocked by the PKC inhibitor GFl09203X, the activation of ERK2 was completely abrogated, along with a detectably enhanced JNK1 activation; FasL upregulation, and apoptosis. However, the FasL upregulation and apoptosis were significantly inhibited by the PKC activator PMA, with a remarkable increase in the ERK2 activation. The cytotoxic effect of aburatubolactam C was reduced in the presence of the anti-Fas neutralizing antibody ZB-4. Although ectopic expression of Bcl-2 failed to completely block the cytotoxicity of aburatubolactam C, it was clearly suppressed. The c-Fos mRNA expression was upregulated in a biphasic manner, where the second phasic expression overlapped with the FasL upregulation. Accordingly, these results demonstrate that aburatubolactam C-induced apoptosis is exerted, at least in part, by FasL upregulation dictated by activation of the PTK ($p56^{lck}\;and\;p59^{fyn}$) /JNKI pathway, which is negatively affected by the concurrent activation of the PKC/ERK2 pathway proximal to PTK activation.

Effect of Storage Times on Sperm Function, Sperm Chromatin Structure Assay (SCSA) and Correlations Between Fertility and SCSA in Boars (액상 정액의 보존 기간이 정자 기능 및 정자염색질 구조 분석에 미치는 영향과 인공수정 분만율과의 상관관계)

  • 유재원;이주형;김인철;이일주;강 권;민동수;윤희진;윤종택;방명걸;류범용;정영채;김창근
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.785-796
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    • 2006
  • This study was designed to evaluate the changes in sperm motility, viability, HOST(hypo-osmotic swelling test), IVP(in vitro penetration), SCSA(sperm chromatin structure assay) during storage of liquid semen collected from boars with different farrowing rates using AI, and to find the relationship between boar fertility through AI and sperm diagnostic parameters during semen storage. The results of HOST were significantly decreased according to the increasing of in semen storage days and the results of IVP were significantly decreased at 3 days of semen storage (P<0.05). The %Red was significantly different among the >80%, 70󰠏80% and <70% farrowing rate group at semen storage day 6(P<0.05). The correlation coefficients between the %Red and farrowing rate were increased according to the semen storage. In conclusion, these results suggest that the sperm parameters evaluated in these studies may be useful indicators to predict the fertility of AI and evaluate the semen quality in boars.

Radioprotective effect of naringin and naringenin against cellular damage and oxidative stress of γ-irradiated mice (감마선을 조사한 마우스의 세포 손상과 산화적 스트레스에 대한 나린진과 나린제닌의 방사선방호 효과)

  • Kang, Jung Ae;Kim, Hye Rim;Yoon, Seon Hye;Jang, Beom-Su;Choi, Dae Seong;Park, Sang Hyun
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.659-667
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    • 2017
  • The present study was designed to evaluate the antioxidant activity and radioprotective effects of Naringin and Naringenin in ${\gamma}$-irradiated mice. The antioxidant activity of Naringin and Naringenin was evaluated by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) assays. Healthy female BALB/c mice were administered Naringin and Naringenin orally ($90{\mu}M/dose$ and $180{\mu}M/dose$) for 7 consecutive days prior to ${\gamma}$-irradiation (6 Gy). Naringenin displayed a much higher antioxidant activity in ABTS and FRAP than naringin. ${\gamma}$-irradiation resulted in cellular damage with decreased spleen and thymus indices and white blood cells (WBC) count. Additionally, ${\gamma}$-irradiation significantly increased lipid peroxidation and decreased the levels of antioxidant enzymes and glutathione (GSH) in the liver tissue. Strikingly, prior administration of Naringenin resulted in considerable prevention of these symptoms. Protection against ${\gamma}$-irradiation-induced cellular damage by Naringenin is likely due to its higher its antioxidant activity. Together, these results confirm that Naringenin is a potent antioxidant and radioprotector.

Effect of Bacteria Eliminated Sperm by Percoll Method on Sperm Quality and Embryo Cleavage in Miniature Pig (Percoll에 의한 미니돼지 정액내 세균 제거가 정자 성상과 수정란 분할에 미치는 영향)

  • Yoo, Han-Jun;Jeon, Jun-Myeong;Lee, Yong-Seung;Cheong, Hee-Tae;Yang, Boo-Keun;Kim, Dae-Young;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.35-40
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    • 2009
  • The objectives of this study was to evaluate the efficiency of the bacteria eliminated sperm by percoll gradient method on sperm quality and embryo cleavage in vitro in pig. The semen of miniature pig collected by gloved-hand method pre-warmed ($37^{\circ}C$) in thermos bottle, and separated by 65% percoll. Analysis of sperm ability was estimated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) and the abnormality. Also, fertility of sperm was monitored with cleavage rate of embryo after IVF using separated and un-separated sperm by percoll. The result, viability of separated sperm was significantly(p<0.05) higher($83.6{\pm}$2.0 vs $59.0{\pm}4.4%$) than un-separated sperm. The results of CTC analysis showed the percentage of F- and B-patterned separated sperm was higher in separated that than un-separated sperm. On the contrary, the percentage of AR-patterned form unseparaed sperm was significantly(p<0.05) higher($13.6{\pm}0.8$ vs $8.1{\pm}0.6%$) than separated sperm. Also, abnormality of un-separated sperm was significantly(p<0.05) higher($2.2{\pm}0.4$ vs $16.8{\pm}2.8%$) than separated sperm. However, the cleavage rates of embryo using separated sperm by percoll and un-separated sperm had not significantly difference on 2 cell stage(9.25 vs 11.88%), 4 cell stage(26.76 vs 24.51%) and >4 cell stage(63.99 vs 63.61%) at 48h of IVF. Therefore, the sperm separated by percoll method showed improvement in sperm quality than un-separated sperm in miniature pig.

Biochemical Effect on Potato Tubers Irradiated by Gamma-Ray at Sprout-Inhibition Dose (방사선 조사에 의한 감자 발아 억제시 생화학적 효과)

  • Jeon, J.H.;Byun, S.M.;Chang, Y.S.P.;Chung, K.H.;Cho, H.O.
    • Applied Biological Chemistry
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    • v.28 no.1
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    • pp.28-35
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    • 1985
  • Potato tubers treated at $4^{\circ}C$ for 4 weeks were irradiated with a dose of 0.12 kGy from $^{60}Co$ source and stored at $20^{\circ}C,\;70{\sim}90%$ humidity for 5 weeks. Changes of ${\alpha}-amylase$, peroxidase, indole acetic acid oxidase, indole acetic acid synthesizing enzyme activities were determined. In addition, treatment of gibberellin or indole acetic acid to tubers irradiated were carried out to examine reversal of sprout-inhibition of tubers irradiated. Results are as follows; 1. Irradiation by ${\gamma}-ray$ at 0. 12 kGy dose inactivated easily the enzyme activities in vitro. $D_{37}$ values obtained were 0.94, 0.36 kGy for ${\alpha}-amylase$ and peroxidase, respectively 2. Complete inhibition of the toter sprouting was resulted by the irradiation of tubers with a dose of 0.12 kGy. 3. The indole acetic acid oxidase activity increased 2 times immediately after irradiation. Meanwhile, indole acetic acid synthesizing activity decreased about $50{\sim}75%$ for 5-week storage in irradiated potatoes, whereas the activity increased about 3.5 times along with sprouting in non-irradiated tubers. 4. In morphological aspects, deformed buds with necrosis in the meristmatic tissue were developed in irradiated tubers. Treatment of gibberellin or indole acetic acid at the concentration of 100 or 20 ppm to the irradiated tubers reversed the sprout-inhibition partially. Nevertheless, the deformed buds remained without change.

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Hypoglycemic and hypolipidemic effects of Saururus chinensis Baill in streptozotocin-induced diabetic rats

  • Hwang, Ji-Yeon;Zhang, Jian;Kang, Min-Jung;Lee, Soo-Kyung;Kim, Hyun-A;Kim, Jong-Jin;Kim, Jung-In
    • Nutrition Research and Practice
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    • v.1 no.2
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    • pp.100-104
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    • 2007
  • Saururus chinensis Baill was reported to inhibit ${\alpha}-glucosidase$ in vitro and flatten postprandial increase in blood glucose in streptozotocin (STZ)-induced diabetic rats. We studied the effect of chronic consumption of S. chinensis Baill on blood glucose and lipid profile in STZ-induced diabetic male rats fed high fat diet. Male rats weighing 100-120 g were fed 30% fat diet with and without 10% freeze-dried leaves of S. chinensis Baill for 7 weeks after 1 week of adaptation. The rats were rendered diabetic by intravenous injection of STZ (60 mg/kg) after 6-week feeding of the assigned diets. At 1 week after the injection, the rats were sacrificed after an overnight fast. Plasma glucose ($380.2{\pm}14.4mg/dL$), total cholesterol ($93.9{\pm}7.9mg/dL$) and triglyceride levels ($123.6{\pm}7.5mg/dL$) of the S. chinensis Baill group were significantly lower than those of the control group ($418.1{\pm}12.0mg/dL,\;119.9{\pm}9.4mg/dL,\;152.0{\pm}10.3mg/dL$, respectively, p<0.05). Chronic consumption of S. chinesis Baill significantly decreased maltase activity of the small intestinal mucosa ($120.1{\pm}8.7U/g$) protein compared with the control group ($96.8{\pm}7.0U/g protein, p<0.05). These results suggest that S. chinensis Baill have hypoglycemic and hypolipidemic effects by inhibiting ${\alpha}-glucosidase$ activity in the animal model of diabetes mellitus.

Effect of acidic electrolyzed water on the quality improvement of boiled-dried anchovy (자건 멸치의 품질 개선에 대한 산성 전해수의 영향)

  • Lee, Soo-Jung;Sung, Nak-Ju;Kang, Shin-Kwon
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.357-364
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    • 2014
  • The quality improvement of the boiled-dried anchovy treated with acidic electrolyzed water (AEW) was investigated. The L value was higher in the anchovy treated with AEW than in the control. Moreover, its ${\Delta}E$ value was 5.08 higher than that of the control. The lipophilic browning degree was also higher than the hydrophilic degree. The degrees of calcium absorption of the control and anchovy treated with AEW were 10.77% and 12.61%, respectively. During their storage in the different conditions (five weeks in $20^{\circ}C$, ten weeks in $4^{\circ}C$, and five months in $-20^{\circ}C$), the lipophilic browning degree of the anchovy treated with AEW was significantly lower than that of the control at the $20^{\circ}C$ storage. The volatile basic nitrogen (VBN) content gradually increased as the storage period increased. During the $20^{\circ}C$ storage, the content of the anchovy treated with AEW was lower than that of the control. The lipid peroxide contents increased with the storage periods, regardless of the temperature, but the content of the anchovy treated with AEW was lower than that of the control. These results indicate that AEW treatment can improve the color quality of boiled-dried anchovy and reduce its VBN and lipid peroxide contents.

Survival of isolated human preantral follicles after vitrification: Analyses of morphology and Fas ligand and caspase-3 mRNA expression

  • Wiweko, Budi;Soebijanto, Soegiharto;Boediono, Arief;Mansyur, Muchtaruddin;Siregar, Nuryati C;Suryandari, Dwi Anita;Aulia, Ahmad;Djuwantono, Tono;Affandi, Biran
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.4
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    • pp.152-165
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    • 2019
  • Objective: This study aimed to examine the effect of vitrification on apoptosis and survival in human preantral follicles after thawing. Methods: This experimental study was conducted at an acute tertiary care hospital from March 2012 to April 2013. Ovaries were sliced into 5 × 5 × 1-mm pieces and divided into the following three groups: preantral follicle isolation, ovarian tissue vitrification-warming followed by follicle isolation, and immunohistochemistry of fresh ovarian tissue. For statistical analyses, the Student t-test, chi-square test, Kruskal-Wallis test, and Kaplan-Meier survival analysis were used. Results: A total of 161 preantral follicles (70% secondary) were collected from ovarian cortex tissue of six women between 30 and 37 years of age who underwent oophorectomy due to cervical cancer or breast cancer. There were no significant differences in the follicular morphology of fresh preantral follicles and vitrified follicles after thawing. The mean Fas ligand (FasL) mRNA expression level was 0.43 ± 0.20 (relative to β-actin) in fresh preantral follicles versus 0.51 ± 0.20 in vitrified follicles (p= 0.22). The mean caspase-3 mRNA expression level in fresh preantral follicles was 0.56 ± 0.49 vs. 0.27 ± 0.21 in vitrified follicles (p= 0.233). One vitrified-thawed secondary follicle grew and developed to an antral follicle within 6 days of culture. Conclusion: Vitrification did not affect preantral follicle morphology or mRNA expression of the apoptosis markers FasL and caspase-3. Further studies are required to establish whether vitrification affects the outcomes of in vitro culture and the maturation of preantral follicles.

A STUDY ON THE ENAMEL EROSION BY CARBONATED BEVERAGE (탄산음료의 법랑질 침식효과에 대한 연구)

  • Lee, Kyung-Ho;Park, Soo-Jin;Jung, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.144-151
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    • 2005
  • Erosion is believed to be the predominant cause of teeth wear in children and young adults, although there will at ways be a contribution from attrition and abrasion. The pH of cola is known to be low and have, therefore, been implicated in the increasing incidence of erosion. The aim of present study was to evaluate the effect of cola on the progression of erosive demineralization in human enamel using demineralization model in vitro. Six groups of human enamel slap were immersed(5 min each bath) in fresh cola, with immersions taking place with or without agitation, and under 3 regimes of frequency intake(low intake, 1 immersion/day; medium, 5/day; high, 10/day). Quantitative assessments of surface erosion were done over an 8-day interval using surface microhardness testing. 1. The average pH of cola was 2.5, which was acidic enough to cause tooth erosion. 2. All the enamel specimen exposed to cola showed erosion like lesions and surface hardness decreased in proportion to the length of immersion (p<0.05). 3. The surface hardness of enamel decreased in proportion to the frequency of immersion (p<0.05). 4. Increased degassing from the drink by gitation accelerated the enamel softening compared with those without agitation.

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