• Title/Summary/Keyword: related genes

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Analysis of flavonoids in double haploid population derived from microspore culture of F1 hybrid of Brassica rapa (배추 종간 잡종의 소포자배양에 의한 Double haploid 집단의 플라보노이드 함량 분석)

  • Seo, Mi-Suk;Won, So Youn;Kang, Sang-Ho;Kim, Jung Sun
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.35-41
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    • 2017
  • One of the most important species, Brassica rapa encompasses a variety of commercial vegetables, such as the Chinese cabbage, pak choi and oilseed crops. The LP08 of yellow sarson (Brassica rapa ssp, tricolaris) have a distinct morphology, with yellow seed color and a unique tetralocular ovary. LP21 of pak choi (Brassica rapa ssp, chinensis) have a dark brown seed color and bilocular ovary. In this study, we generated double haploid plants by crossing the LP08 (maternal variety) and LP21 (paternal variety), using microspore culture. A total of 66 accessions with various morphological characteristics were used for content analysis of flavonoids. The three flavonoids, quercetin, naringenin and kaempferol, showed differing contents in the two crossing parents. The Chinese cabbage type 'Chiifu' was used as the control. The highest accumulation of total flavonoids was observed in LP08. The lowest mean total flavonoids were found in 'Chiifu'. Among the 66 DH accessions, the quercetin contents of 18 accessions showed higher content than LP08. Kaempferol content was also high, and was found to be 79.7% of the total flavonoid content. Naringenin content was low at 2.8%, and was not detected in 22 accessions. Interestingly, the quercetin content positively correlated with the kaempferol content. These results can be used to identify genetic locus and genes related to useful traits. Phenotypic analysis of 66 DH accessions can further be used for natural selection of good breeding materials in B. rapa.

Evaluation of primary hepatocyte function using 2D or 3D culture method for primary rat hepatocytes (Rat Primary Hepatocyte의 2차원 배양과 3차원 배양에 따른 생리 활성능과 대사능에 관한 연구)

  • Lim, Malgum;Kim, Yeongji;Shin, Yurianna;Oh, Keon Bong;Hwang, Seongsoo;Kim, Youngim;Hur, Tai-Young;Ock, Sun A
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.169-177
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    • 2016
  • There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy 3.5×106 primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

Heat Shock Protein Augmentation of Angelica gigas Nakai Root Hot Water Extract on Adipogenic Differentiation in Murine 3T3-L1 Preadipocytes

  • Lumbera, Wenchie Marie L.;Cruz, Joseph dela;Yang, Seung-Hak;Hwang, Seong Gu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.419-427
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    • 2016
  • There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at 42C for one hour and then allowed to recover at normal incubation temperature of 37C for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to 400μg/mL) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

Expression of Superoxide Dismutase Isoenzyme Genes and Enzyme Activities in Rice Irradiated with a High-Dose Gamma Ray (고선량 감마선을 조사한 벼에서 SOD isoenzyme들의 유전자 발현 및 효소활성)

  • Chae Hyo-Seok;Kim Jin-Hong;Chung Byung-Yeoup;Kim Jae-Sung;Wi Seung-Gon;Baek Myung-Hwa;Cho Jae-Young
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.180-185
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    • 2006
  • We investigated relations between physiological damages and gene expression and enzyme activities of superoxide dismutase(SOD) isoenzymes in leaves of rice (Oriza sativa L. cv. Ilpoombyeo) plants irradiated with a high-dose gamma-ray. Gamma-irradiation with 500 Gy caused significant decreases in the contents of protein, chlorophyll and carotenoid in the rice leaves by 24 h, especially reducing the chlorophyll contents up to 26% relative to the control. In contrast, gene expressions of SOD isoenzymes were kept higher in the irradiated leaves until 24 h after the irradiation than in the control and they started to noticeably decrease at 48 h, finally being lower in the irradiated leaves at 72 h than in the control. In the case of enzyme activities of SOD isoenzymes, some CuZn-SOD isoenzymes showed slightly increased activities until 48 h after the irradiation but at 72 h, all isoenzyme activities markedly decreased in the irradiated leaves below the control levels. In conclusion, 500 Gy gamma-irradiation used in the current study caused decreases in the contents of protein, chlorophyll and carotenoid as symptoms for physiological damages. Although such physiological damages were not directly related to the gene expressions and enzyme activities of SOD isoenzymes until 24 h after the irradiation, the damages at 72 h were reasonably attributable to their reduction.

Identification of Amino Acids Involved in the Sensory Function of the PrrB Histidine Kinase by Site-directed Mutagenesis (Site-directed mutagenesis에 의한 PrrB histidine kinase의 신호인지 기능에 관련된 아미노산의 발굴)

  • Kim Yong-Jin;Ko In-Jeong;Oh Jeong-Il
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.485-492
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    • 2006
  • The PrrBA two-component system is one of the major regulatory systems that control expression of photosynthesis genes in response to changes in oxygen tension in the anoxygenic photosynthetic bacterium, Rhodobacter sphaeroides. The system consists of the PrrB histidine kinase and the PrrA response regulator. The N-terminal transmembrane domain of PrrB serves as a signal-sensing domain and comprises six transmembrane helices forming three periplasmic loops and two cytoplasmic loops. The 3rd and 4th transmembrane helices and the 2nd periplasmic loop were suggested to play a crucial role in redox-sensory function. In this study we demonstrated that mutations of Asp-90, Gln-93, Leu-94, Leu-98, and Asn-106 in the 2nd periplasmic loop and its neighboring region led to severe defects in PrrB sensory function, indicating that these amino acids might be related to the redox-sensing function of PrrB. The mutant forms (D90E, D90N, and D90A) of PrrB were heterologously overexpressed in Escherichia coli, purified by means of affinity chromatography and their autokinase activities were comparatively assessed. The D90N form of PrrB was shown to possess higher autokinase activity than the wild-type form of PrrB, whereas the D90E form of PrrB displayed lower autokinase activity than the wild-type form of PrrB. The D90A mutation led to the loss of PrrB autokinase activity.

Changes in expression of monocarboxylate transporters, heat shock proteins and meat quality of Large White Yorkshire and Ghungroo pigs during hot summer period

  • Parkunan, Thulasiraman;Das, Arun K.;Banerjee, Dipak;Mohanty, Niharika;Paul, Avishek;Nanda, P.K.;Biswas, TK;Naskar, Syamal;Bag, Sadhan;Sarkar, Mihir;Mohan, Narayana H.;Das, Bikash Chandra
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.246-253
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    • 2017
  • Objective: Present study explores the effect of hot summer period on the glycolytic rate of early post-mortem meat quality of Ghungroo and Large White Yorkshire (LWY) pig and comparative adaptability to high temperature between above breeds by shifting the expression of stress related genes like mono-carboxylate transporters (MCTs) and heat shock proteins (HSPs). Methods: Healthy pigs of two different breeds, viz., LYW and Ghungroo (20 from each) were maintained during hot summer period (May to June) with a mean temperature of about 38C. The pigs were slaughtered and meat samples from the longissimus dorsi (LD) muscles were analyzed for pH, glycogen and lactate content and mRNA expression. Following 24 h of chilling, LD muscle was also taken from the carcasses to evaluate protein solubility and different meat quality measurements. Results: LWY exhibited significantly (p<0.01) higher plasma cortisol and lactate dehydrogenase concentration than Ghungroo indicating their higher sensitivity to high temperature. LD muscle from LWY pigs revealed lower initial and ultimate pH values and higher drip loss compared to Ghungroo, indicating a faster rate of pH fall. LD muscle of Ghungroo had significantly lower lactate content at 45 min postmortem indicating normal postmortem glycolysis and much slower glycolytic rate at early postmortem. LD muscle of LWY showed rapid postmortem glycolysis, higher drip loss and higher degrees of protein denaturation. Ghungroo exhibited slightly better water holding capacity, lower cooking loss and higher protein solubility. All HSPs (HSP27, HSP70, and HSP90) and MCTs (MCT1, MCT2, and MCT4) in the LD muscle of pigs inclined to increase more in Ghungroo than LWY when exposed to high temperature. Conclusion: Effect of high temperature on the variation of HSPs and MCTs may play a crucial role in thermal tolerance and adaptation to different climatic conditions, pH regulation, muscle acidification, drip loss, protein denaturation and also in postmortem meat quality development.

Effects of Unripe Black Raspberry Extracts on Prostate Cancer Cell Line and Rat Model of Benign Prostatic Hyperplasia (복분자 미숙과 추출물이 전립선암 세포주와 전립선비대 백서모델에 미치는 영향)

  • Lee, Su Jung;Choi, Hye Ran;Lee, Jung-Hyun;Kwon, Ji Wung;Lee, Hee Kwon;Jeong, Jong Tae;Lee, Tae-Bum
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.507-515
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    • 2014
  • Benign prostatic hyperplasia (BPH), which is commonly found in aging men, is characterized by hyperplasia of prostatic stromal and epithelial cells beginning in the periurethral zone of the prostate. The prevalence of BPH increases in an age-dependent manner. Here, we investigated the protective effects of unripe Rubus occidentalis extracts (UROE) on BPH development using a prostate cancer cell line and testosterone-induced BPH rat model. Experiments using an established hormone-dependent prostate cancer cell line (LNCaP) showed that UROE treatment significantly decreased expression of androgen-related genes, including androgen receptor (AR), prostate specific antigen (PSA), and 5-alpha reductase 2, but not 5-alpha reductase 1, which was also observed in flutamide-treated cells. Further, AR and PSA gene expression was reduced by UROE treatment under androgen-stimulated conditions using dihydrotestosterone (DHT). BPH animals displayed elevated prostate weights. However, UROE as well as finasteride treatment significantly reduced prostate weights and DHT levels compared to testosterone-induced BPH animals. Histopathological analysis also showed that UROE treatment suppressed testosterone-induced prostatic hyperplasia. Taken together, the results suggest that UROE may effectively inhibit the development of BPH and thus may be a useful agent in BPH treatment.

Inhibitory effect of Mojaban (Sargassum fulvellum) extract on ultraviolet B-induced photoaging in hairless mouse skin (무모 마우스 피부에서 모자반 추출물의 자외선비(ultraviolet B)에 의해 유도된 광노화 억제 효과)

  • Rhim, Dong-Bin;Kim, Changhee;Hwang, Jae-Kwan
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.467-475
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    • 2020
  • Ultraviolet radiation is a major cause of photoaging, characterized by wrinkle formation and skin dehydration. Mojaban (Sargassum fulvellum), belonging to the brown algae of the Sargassaceae family, exerts anti-inflammatory, antioxidant, and anti-atopic activities. Particularly, Mojaban extract alleviated ultraviolet B (UVB)-induced inflammatory responses in human keratinocytes and BALB/c mice. Here, the dorsal skin of hairless mice was exposed to UVB for photoaging. The mice were administrated 300 mg/kg Mojaban extract (S. fulvellum extract; SFE) daily. Phenotypically, SFE reduced wrinkle formation and epidermal thickness and increased skin elasticity. SFE increased hydroxyproline content by upregulating the expression of collagen-related genes and downregulating matrix metalloproteinase expression. Furthermore, SFE increased skin moisture content and inhibited transepidermal water loss. SFE upregulated the expression of involucrin, loricrin, and transglutaminase for cornified envelope formation as well as the expression of filaggrin and caspase-14 for natural moisturizing factors. Collectively, SFE may be employed as a nutraceutical ingredient for anti-photoaging.

Endogenous Nitric Oxide Strengthens Doxorubicin-induced Apoptosis in Human Colorectal Cell Lines (Doxorubicin에 의한 내인성 산화질소가 인간 대장암 세포주에서의 세포사멸에 미치는 효과)

  • Im, Soon Jae;Kim, Ji Hye;Kim, Min Young
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1137-1143
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    • 2014
  • Doxorubicin is a general chemotherapy drug widely used for a number of cancers. However, the correlation between endogenous nitric oxide (NO) levels and chemoresistance to doxorubicin remains unclear. In this study, we investigated the effect of endogenous NO on the anticancer activity of doxorubicin in human colon cancer cell lines HCT116 and HT29 with different p53 status. The cells were treated with either doxorubicin alone or in combination with the NO synthase (NOS) inhibitor NG-monomethyl-L-arginine (NMA). Doxorubicin differentially inhibited the growth of both the HCT116 (p53-WT) and HT29 (p53-MUT) cells, which was mitigated by cotreatment with NMA. Further studies revealed that inhibition of endogenous NO mitigated doxorubicin-induced apoptosis in the HCT116 and HT29 cells, as evidenced by apoptotic DNA fragmentation and the sub-G1 peak of apoptotic markers. Apoptosis was delayed in the HT29 cells, and its magnitude was greatly reduced, underscoring the importance of the modulation of p53 in the response. RT-PCR analysis revealed that doxorubicin down-regulated levels of inhibitors of the apoptosis family (cellular IAP-1 and-2). Collectively, these data show that induction of apoptosis by doxorubicin in human colon cancer cells is possibly related to modulation of endogenous NO, the expression of the IAP family of genes, and the status of p53. The underlying mechanisms may represent potential targets for adjuvant strategies to improve the efficacy of chemotherapy for colon cancer.

Effects of Platycodon grandiflorum on the Induction of Autophagy and Apoptosis in HCT-116 Human Colon Cancer Cells (길경 추출물에 의한 HCT-116 대장암 세포주에서의 autophagy와 apoptosis 유발 효과)

  • Hong, Su Hyun;Park, Cheol;Han, Min Ho;Kim, Hong Jae;Lee, Moon Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1244-1251
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    • 2014
  • Platycodon grandiflorum (PG) has been known to possess many biological effects, including anti-inflammatory and anti-allergy activity and anti-obesity and hyperlipidemia effects. However, little research has been conducted regarding its anticancer effects, with the exception of its ability to stimulate apoptosis in skin cells. There has also been no study regarding PG-induced autophagy. The modulation of autophagy is recognized as one of the hallmarks of cancer cells. Depending on the type of cancer and the context, autophagy can suppress or help cancer cells to overcome metabolic stress and the cytotoxicity of chemotherapy. Therefore, the present study was designed to investigate whether or not extracts from PG-induced cell death were connected with autophagy and apoptosis in HCT-116 human colon cancer cells. PG stimulation decreased cell proliferation in a dose- and time-dependent manner and induced apoptosis, which was partially dependent on the activation of caspases. PG treatment also resulted in the formation of autophagic vacuoles simultaneously with regulation of autophagy-related genes. Interestingly, a PG-mediated apoptotic effect was further triggered by pretreatment with the autophagy inhibitors 3-methyladenin and bafilomycin A1. However, cell viability recovered quite well with bafilomycin A1 treatment. These findings show that PG treatment promotes both autophagy and apoptosis and that PG-induced autophagic response might play a role in the autophagic cell death of HCT-116 cells.