• Animal cells and systems
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• v.9 no.4
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• pp.191-198
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• 2005

The effects of 6-aminonicotinamide (6-AN) on viability of IMR32 neuroblastoma cells in the presence of ATP or $NAD^+$ have been investigated. 6-AN caused marked reduction in cell viability and similar observations were also made with cells treated with 6-AN+ATP. However, cells treated with $6-AN+NAD^+$ showed cell viability similar to untreated cells. Morphologically, 6-AN and 6-AN+ATP treated cells showed loss of neurites, polyhedric shapes, shrinkage of cell bodies and formation of lysed cells, while $6-AN+NAD^+$ cells did not show any such changes. The flow cytometry analysis demonstrated that 6-AN increased cell population in $G_0/G_1$ phase and decreased cell population in Sand $G_2/M$ phase following a 72 h exposure. Western blot analysis showed that 6-AN stimulated a substantial increase in the level of the cdk inhibitor $p27^{kip1}$, but lowered the levels of cdk2, cyclin E and p-Rb. However, cdc25A and p53R2 were not significantly affected. Immunofluorscence staining of $p27^{kip1}$, cdk2, cyclin E and p-Rb revealed close correlation between the signal observed in the Western blot analysis. 6AN+ATP treated cells showed similar results obtained with 6-AN treated cells in expression of cdk2, cyclin E, p-Rb proteins and $p27^{kip1}$, $6-AN+NAD^+$ cells showed greater expression of cdk2, cyclin E and p-Rb than those in 6-AN and 6-AN+ATP treated cells. The results suggest that 6-AN induced the $G_0/G_1$ phase arrest in IMR32 neuroblastoma cell lines through the increase of $p27^{kip1}$ and the decrease of cdk2, cyclin E and p-Rb.

• Journal of Life Science
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• v.13 no.5
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• pp.642-650
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• 2003

To develop a new approach to the treatment of neuroblastoma cells we evaluated the effect of cAMP on the Ewing's sarcoma cell line CHP-100. We observed that the proliferation-inhibitory effect of cAMP analogs was due to cell cycle arrest and induction of apoptosis, which was confirmed by observing the morphological changes and DNA fragmentation. DNA flow cytometric analysis revealed that cAMP arrested the cell cycle progression at the G1 phase, which effects were associated with inhibition of phosphorylation of retinoblastoma protein (pRB) and enhanced binding of pRB and the transcription factor E2F-1. cAMP also suppressed the cyclin-dependent kinase (Cdk) 2 and cyclin E-associated kinase activity without changes of their expressions. Furthermore, cAMP induced the levels of Cdk inhibitor $p21^{WAF1/CIP1$ expression and p21 proteins induced by cAMP were associated with Cdk2. Overall, our results identify a combined mechanism involving the inhibition of pRB phosphorylation and induction of p21 as targets for cAMP, and this may explain some of its anti-cancer effects.

• Asian-Australasian Journal of Animal Sciences
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• v.1 no.4
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• pp.201-208
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• 1988

Fifty-six lactating Surti buffaloes, fed rice straw, were allocated to seven treatment groups as follows: 1. Straw supplemented with 2% urea (SS) + 1.5kg rice bran (RB) 2. Straw treated with 4% urea in an open stack (TS open) 3. TS open + 1.5 kg RB 4. TS open + 3.0 kg RB 5. TS open + 1.5 kg RB + 3.0 kg Gliricidia (Gl) 6. Straw treated with 4% urea in a closed pit (TS closed) 7. TS closed + 1.5 kg RB + 3.0 kg Gl Milk production, butterfat percentage and liveweight gain of cows and calves were measured and tested with analysis of variance. The results are: - The animals on urea treated straw (group 2) had a higher milk production (p<0.05), higher butterfat production (p<0.05) and less liveweight gain loss (p<0.05) than the animals on urea supplemented straw (group 1). Butterfat percentage also increased by treatment, although not significantly (p>0.05). - Increasing levels of rice bran (groups 3 and 4 compared to 2) increased total milk production and milked quantity of butterfat, while butterfat percentage decreased (p < 0.05). - Milk production increased (p <0.05) with extra rice bran added (group 4 compared to 3), but was not affected (p > 0.05) by Gliricidia addition (group 5 compared to 3). Butterfat percentage dropped with extra rice bran supplement (p <0.05). The lack of response to Gliricidia indicated that protein is not limiting in treated straw, or that Gliricidia protein is partly insoluble. - System of treatment had no effect on milk production (p >0.05), while supplementation with 1.5 kg RB and 3.0 kg Gliricidia increased production and caused a lower butterfat percentage (p <0.05) (groups 2, 5, 6 and 7 compared). A significant (p <0.05) interaction treatment system x supplementation was present. It was concluded, that both treatment and supplementation did affect milk production as well as milk composition. Gliricidia addition gave less effect than rice bran, indicating different requirements for starchy substances in the feed. Treatment of straw does not negatively affect butterfat production, it can increase butterfat production and even butterfat percentage.

• The Korean Journal of Food And Nutrition
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• v.23 no.2
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• pp.233-239
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• 2010

Phenolic acid concentrates of rice bran(RB-ex) and hydroxycinnamic acids were investigated for their anti-hyperglycemic activities through glucose uptake and glucokinase activity using HepG2 cells and stimulatory effects on insulin secretion using HIT-T15 cells. RB-ex was prepared as an ethylacetate extract after alkaline hydrolysis and hydroxycinnamic acids, found as major compositions of RB-ex, such as ferulic acid(FA), sinapic acid(SA) and p-coumaric acid(p-CA) were investigated to compare with the properties of RB-ex. The properties of glucose uptake in HepG2 cells were examined in the absence of insulin and two different glucose concentrations(5.5 mM and 25 mM). RB-ex and FA showed anti-hyperglycemic activities through the increase of glucose uptake and the stimulation of glucokinase activity in HepG2 cells. RB-ex exhibited higher glucose uptakes with higher glucose concentrations, whereas FA exhibited the same increasing effects on both concentrations of glucose. RB-ex and FA exhibited doubled glucokinase activities relative to control. In the presence of insulin in the 25 mM glucose-containing medium, the levels of glucose uptake were increased in all treatments compared with control. As stimulatory effects of samples on insulin secretion were estimated, RB-ex and FA stimulated insulin secretion at a concentration of 25 ${\mu}g/m{\ell}$ and in particular, FA showed the highest amount of insulin-release in HIT-T15 cells. Antioxidative effects on HIT-T15 cells, RB-ex and hydroxycinnamic acids, excluding p-CA, showed inhibitory activities of 78% to 80% at a concentration of 100 ${\mu}g/m{\ell}$. On the basis of these results, we conclude that RB-ex and FA could help decrease blood glucose levels and prevent the cell damages via antioxidant activity.

• Bulletin of the Korean Chemical Society
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• v.19 no.1
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• pp.98-103
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• 1998

The crystal structure of dehydrated $Rb^+$-exchanged zeolite X, stoichiometry $Rb_{71}Na_{21}-X\; (Rb_{71}Na_{21}Si_{100}Al_{92}O_{384})$ per unit cell, has been determined from single-crystal X-ray diffraction date gathered by counter methods. The structure was solved and refined in the cubic space group Fd3, a=25.007(3) Å at 21(1) ℃. The crystal was prepared by ion exchange in a flowing stream using a 0.05 M aqueous RbOH solution (pH=12.7). The crystal was then dehydrated at 360 ℃ and $2{\times}10^{-6}$ torr for two days. The structure was refined to the final error indices, $R_1=0.047$ and $R_2=0.040$ with 239 reflections for which I> 3σ(I). In this structure, 71 $Rb^+$ ions per unit cell are found at six different crystallographic sites and 21 $Na^+$ ions per unit cell are found at two different crystallographic sites. Four and a half $Rb^+$ ions are located at site Ⅰ, the center of the hexagonal prism. Nine $Rb^+$ ions are found at site Ⅰ' in the sodalite cavity (Rb-O=2.910(15) Å and O-Rb-O=78.1(4)°). Eighteen $Rb^+$ ions are found at site Ⅱ in the supercage (Rb-O=2.789(9) Å and O-Rb-O=92.1(4)°). Two and a half $Rb^+$ ions, which lie at site Ⅱ', are recessed ca. 2.07 Å into the sodalite cavity from their three O(2) oxygen planes (Rb-O=3.105(37) Å and O-Rb-O=80.6(5)°). Thirty-two $Rb^+$ ions are found at site Ⅲ deep in the supercage (Rb-O=2.918(12) Å and O-Rb-O=71.9(4)°), and five $Rb^+$ ions are found at site Ⅲ'. Seven $Na^+$ ions also lie at site Ⅰ. Fourteen $Na^+$ ions are found at site Ⅱ in the supercage (Na-O=2.350(19) Å and O-Na-O=117.5(6)°).

• BMB Reports
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• v.36 no.2
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• pp.223-229
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• 2003

The product of a tree (Tabebuia avellanedae) from South America, $\beta$-lapachone, is known to exhibit various pharmacological properties, the mechanisms of which are poorly understood. The aim of the present study was to further elucidate the possible mechanisms by which $\beta$-lapachone exerts its anti-proliferative action in cultured human prostate cancer cells. We observed that the proliferation-inhibitory effect of $\beta$-lapachone was due to the induction of apoptosis, which was confirmed by observing the morphological changes and cleavage of the poly(ADP-ribose) polymerase protein. A DNA flow cytometric analysis also revealed that $\beta$-lapachone arrested the cell cycle progression at the G1 phase. The effects were associated with the down-regulation of the phosphorylation of the retinoblastoma protein (pRB) as well as the enhanced binding of pRB and the transcription factor E2F-1. Also, $\beta$-lapachone suppressed the cyclindependent kinases (Cdks) and cyclin E-associated kinase activity without changing their expressions. Furthermore, this compound induced the levels of the Cdk inhibitor $p21^{WAF1/CIP1}$ expression in a p53-independent manner, and the p21 proteins that were induced by $\beta$-lapachone were associated with Cdk2. $\beta$-lapachone also activated the reporter construct of a p21 promoter. Overall, our results demonstrate a combined mechanism that involves the inhibition of pRB phosphorylation and induction of p21 as targets for $\beta$-lapachone. This may explain some of its anticancer effects.

• Molecules and Cells
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• v.45 no.4
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• pp.216-230
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• 2022

SERTA domain-containing protein 1 (Sertad1) is upregulated in the models of DNA damage and Alzheimer's disease, contributing to neuronal death. However, the role and mechanism of Sertad1 in ischemic/hypoxic neurological injury remain unclear. In the present study, our results showed that the expression of Sertad1 was upregulated in a mouse middle cerebral artery occlusion and reperfusion model and in HT22 cells after oxygen-glucose deprivation/reoxygenation (OGD/R). Sertad1 knockdown significantly ameliorated ischemia-induced brain infarct volume, neurological deficits and neuronal apoptosis. In addition, it significantly ameliorated the OGD/R-induced inhibition of cell viability and apoptotic cell death in HT22 cells. Sertad1 knockdown significantly inhibited the ischemic/hypoxic-induced expression of p-Rb, B-Myb, and Bim in vivo and in vitro. However, Sertad1 overexpression significantly exacerbated the OGD/R-induced inhibition of cell viability and apoptotic cell death and p-Rb, B-Myb, and Bim expression in HT22 cells. In further studies, we demonstrated that Sertad1 directly binds to CDK4 and the CDK4 inhibitor ON123300 restores the effects of Sertad1 overexpression on OGD/R-induced apoptotic cell death and p-Rb, B-Myb, and Bim expression in HT22 cells. These results suggested that Sertad1 contributed to ischemic/hypoxic neurological injury by activating the CDK4/p-Rb pathway.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.213-219
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• 2003

We investigated the effects of Insamsapye-tang (ISSPT) water extract on the cell proliferation of human lung carcinoma A549 cells. ISSPT treatment resulted in the inhibition of cell proliferation in a concentration-dependent manner. This anti-proliferative effect of A549 cells by ISSSPT treatment was associated with morphological changes such as membrane shrinking and cell rounding up. DNA flow cytometric histograms showed that population of G1 phase of the cell cycle was increased by ISSPT treatment in a concentration-dependent manner. ISSPT treatment induced the levels of tumor suppressor p53 protein and cyclin-dependent kinase (Cdk) inhibitor p27 without significant alteration of cyclins and Cdks expression. In addition, ISSPT treatment resulted in down-regulation of phosphorylated retinoblastoma protein (pRB). However, the levels of p130, the pRB family protein, and transcription factors. E2F-1 and E2F-4. were remained unchanged. The present results indicated that ISSPT-induced inhibition of lung cancer cell proliferation is associated with the blockage of G1/S progression and the induction of apoptosis, and we suggest that ISSPT will be an effective therapeutic agent on human lung cancer.

• Journal of Life Science
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• v.19 no.8
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• pp.1177-1182
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• 2009

In order to establish in vitro infection model for research of plant pathogen based on tissue softening disease in napa cabbage, eighty independent bacterial strains were isolated from the softened napa cabbage tissues. Eight bacterial isolates were primarily screened with the generation of reproducible tissue softening disease to fresh napa cabbages within 24${\sim}$48 hours after inoculation. Through various microbiological biochemical and morphological examinations, three Gram (-) isolates which harbor independent biological properties were finally chosen, and named as RBI, RB2 and RB6. Collective results obtained from API 20E test and analyses of VITEK 2 COMPACT and nucleotide sequences of 165 rRNA of each isolate proposed that isolates RBI and RB2 are close to the Erwinia carotovora subsp. odorifera, and RB6 is close to the Erwinia carotovora subsp. carotovora. These isolates grew optimally at $30^{\circ}C$ with neutral pH culture condition. The isolates caused softening tissue disease with dose-dependent manner regardless of pre-surface damages of napa cabbage. Minimum dose to cause soft rot disease for RBI, RB2 or RB6 were $8.0{\times}10^8$ CFU/mt $10^9$ CFU/ml or $4.7{\times}10^6$ CFU/ml respectively. These isolates caused tissue softening disease to eggplant, paprika and napa cabbage out of 14 different tested vegetables, indicating that these isolates damages specific plant tissues. The bacterial isolates obtained in this research and in vitro plant infection model will be adapted in the understanding of the mechanism of pathogenesis by plant pathogen.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.177-184
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• 2005

This study was conducted to investigate the effects of recombinant bovine somatotropin (rbST) injection on conception and parturition rates in normal or repeat breeding Hanwoo. We treated 462 cows containing 79 repeat-breeding cows of multiparous and allocating 5 treatment groups. Treatment 1 (T1) was injection of 2ml saline (for pseudo treatment), T2 was one injection of rbST 250mg into the tailhead region at the estrus, T3 was twice injection of rbST 250mg both at the time of insemination and again 10 to 14 day later, T4 was once injection of rbST 500mg at insemination and T5 was twice injection of 500mg rbST both at the time of insemination and again 10 to 14day later respectively. In rbST treated groups, timed artificial inseminations (TAI) were performed fellowing estrus synchronization. 100 us GnRH was injected into the scapula region on Day 0, 25mg $PGF_2{\alpha}$ was injected on Day 7 for degeneration of corpus luteum (CL) and 100ug GnRH was injected for inducing the synchronization. The results are as fellows; When normal Hanwoo were inseminated once with rbST administration, the pregnancy rate of T2 $(67.5\pm18.48\%)$ were higher than control $(52.4\pm9.72\%)$, while the pregnancy rate of T4 $(63.3\pm5.77\%)$ were significantly higher (p.<0.05) than control $(39.3\pm12.89\%)$ in repeat breeder Hanwoo. The parturition rates of normal Hanwoo were no differences among the treatments but were significant different in repeat breeder Hanwoo (p<0.05). When the estrous was induced by Ovsynch and inseminated once with rbST administration, the pregnancy rates of T2 was $12.5\%$ higher than control in normal Hanwoo, T4 $(80.0\%)$ was highest among the treatments (p<0.05) in repeat breeder Hanwoo. When normal Hanwoo were inseminated once with rbST administration, the pregnant period was $282.7\~284.8$ days and the body weight was $25.1\~25.9kg$, there were no difference among the treatments. The ratio of sex was almost same without T4 (male vs. female=18:9). In repeat breeder Hanwoo, pregnant period was 280.4~289.3 day and body weight was $23.0\~26.6kg$, it had no difference among the treatments. The sex ratio were similar to normal Hanwoo except T4 (M : F=2 : 8). In conclusion, the pregnancy and parturition rate by once insemination could be improved by the administration of rbST 250mg in normal Hanwoo or 500mg in repeat breeder Hawoo.