• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8411-8418
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• 2016

The effect of Eruca sativa seed extract (SE) on nuclear factor kappa B (NF-${\kappa}B$), cyclooxygenase-2 (COX-2) and B-cell lymphoma-2 (Bcl-2) gene expression levels was investigated in rat mammary gland carcinogenesis induced by 7,12 dimethylbenz(${\alpha}$)anthracene (DMBA). DMBA increased NF-${\kappa}B$, COX-2 and Bcl-2 gene expression levels and lipid peroxidation (LP), while, decreased glutathione-S-transferase (GST) and superoxide dismutase (SOD) activities and total antioxidant concentration (TAC) compared to the control group. After DMBA administration, SE treatment reduced NF-${\kappa}B$, COX-2 and Bcl-2 gene expression levels and LP. Hence, SE treatment reduced inflammation and cell proliferation, while increasing apoptosis, GST and SOD activities and TAC. Analysis revealed that SE has high concentrations of total flavonoids, triterpenoids, alkaloids and polyphenolic compounds such as gallic, chlorogenic, caffeic, 3,4-dicaffeoyl quinic, 3,5-dicaffeoyl quinic, tannic, cinnamic acids, catechin and phloridzin. These findings indicate that SE may be considered a promising natural product from cruciferous vegetables against breast cancer, especially given its high antioxidant properties.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.269-278
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• 2009

Purpose: The aim of this study was to investigate matrix metalloproteinase 1 (MMP-1) gene polymorphism (1G/2G at -1607 and A/G at -519) in Korean subject and to assess the association between polymorphism and periodontal status. Methods: Forty nine generalized aggressive periodontitis (GAP) patients and 57 periodontally healthy children were recruited and genomic DNA was extracted from buccal swab. The polymorphisms of MMP-1 promoter genes were determined by polymerase chain reaction and restriction fragment length product (PCR-RFLP) method. The distribution of genotype and allele frequency was compared between 2 groups by ${\chi}^2$ test. Results: There was a significant difference in the distribution of genotypes and frequency of alleles between the GAP and reference groups at the position - 519 of MMP-1 gene promoter (P<0.05). Allele G carrier rate was significantly lower in GAP group than that of the reference group (P< 0.001). At the position -1607 of MMP-1 gene promoter, genotype distribution and allele frequency showed no statistically significant difference between the groups. However, in the female group, a significant difference was observed between the groups for the genotype distribution, allele frequency and allele 1G carrier rate (P< 0.05). Conclusions: The DNA polymorphism at the MMP-1 gene promoter might be associated with GAP in Korean.

• KSBB Journal
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• v.29 no.3
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• pp.155-164
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• 2014

Succinic acid, a representative biomass-derived platform chemical, is a major fermentation product of Actinobacillus succinogenes. It is well known that carbon dioxide is consumed during the succinate fermentation, but the biochemical mechanism behind this phenomenon is not yet understood well. In this study, it was found that the addition of carbonic anhydrase (CA)s into media significantly enhances the succinic acid production by A. succinogenes during the fermentation supplied with carbon dioxide. It is likely that the (bi) carbonate produced by the CA activity from gaseous carbon dioxide is favoured by A. succinogenes for consumption and utilization. Therefore, the $MgCO_3$ requirement could be significantly reduced without compromising the succinate productivity. Furthermore, because of too high price of the commercial carbonic anhydrase, it was undertaken to economically overproduce a cyanobacterial carbonic anhydrase by the use of a recombinant Pichia pastoris. An expression vector system was constructed with the carbonic anhydrase gene PCR-cloned from Cyanobacterium Synechocystis sp., and introduced into P. pastoris for fermentation studies. About 95.9 g/L of succinic acid was produced in the production medium with 30 ppm of carbonic anhydrase, approximately 2 fold higher productivity compared to the parallel process with no supplementation of the enzyme. It is expected that this method can provide a valuable way of overcoming inefficiencies inherent in gas supply during $CO_2$-based bioprocesses like succinic acid fermentation.

• Parasites, Hosts and Diseases
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• v.42 no.2
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• pp.85-89
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• 2004

Cryptosporidium parvum oocysts were isolated from a child suffering from acute gastroenteritis and successfully passaged in a calf and mice (designated hereafter SNU-H1) in the Republic of Korea; its molecular genotype has been analyzed. The GAG microsatellite region was amplified by a polymerase chain reaction (PCR), with a 238 base pair product, which is commonly displayed in C. parvum. The isolate was shown to be a mixture of the genotypes 1 (anthroponotic) and 2 (zoonotic). To study its infectivity in animals, 2 calves and 3 strains of mice were infected with the SNU-H1; in these animals, the propagation of both genotypes was successful. In immunosup-pressed (ImSP) BALB/c and C57BU6 mice the number of oocysts decreased after day 10 post-infection (PI); but in ImSP ICR mice, they remained constant until day 27 PI. The results show that both the C. parvum genotypes 1 and 2 can be propagated in calves and ImSP mice.

• Natural Product Sciences
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• v.19 no.4
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• pp.347-354
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• 2013

Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3'-deoxyadenosine). Cordycepin has many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. However, the therapeutic mechanism has not yet been elucidated. In this study, we examined the effects of cordycepin on the antigen-presenting function of antigen-presenting cells (APCs). Dendritic cells (DCs) were cultured in the presence of cordycepin and then allowed to phagocytose microspheres containing ovalbumin (OVA). After washing and fixing, the efficacy of OVA peptide presentation by DCs was evaluated using CD8 and CD4 T cells. Also, we confirmed the protein levels of proinflammatory cytokines through RT-PCR and Western blot analysis. Cordycepin decreased both MHC class I and class II-restricted presentation of OVA and suppressed the expression of both MHC molecules and the phagocytic activity toward exogenous OVA. The class II-restricted OVA presentation-regulating activity of cordycepin was also confirmed using mice that had been injected with cordycepin followed by soluble OVA. Furthermore, cordycepin suppressed the mRNA and protein levels of iNOS, COX-2, pro-inflammatory cytokines in a concentration-dependent manner. These results provide an understanding of the mechanism of the T cell response-regulating activity of cordycepin through the inhibition of MHC-restricted antigen presentation in relation to its actions on APCs.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.17
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• pp.7935-7941
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• 2015

The aim of this study was to implement massive parallel sequencing (MPS) technology in clinical genetics testing. We developed and tested an amplicon-based method for resequencing the BRCA1 and BRCA2 genes on an Illumina MiSeq to identify disease-causing mutations in patients with hereditary breast or ovarian cancer (HBOC). The coding regions of BRCA1 and BRCA2 were resequenced in 96 HBOC patient DNA samples obtained from different sample types: peripheral blood leukocytes, whole blood drops dried on paper, and buccal wash epithelia. A total of 16 random DNA samples were characterized using standard Sanger sequencing and applied to optimize the variant calling process and evaluate the accuracy of the MPS-method. The best bioinformatics workflow included the filtration of variants using GATK with the following cut-offs: variant frequency >14%, coverage ($>25{\times}$) and presence in both the forward and reverse reads. The MPS method had 100% sensitivity and 94.4% specificity. Similar accuracy levels were achieved for DNA obtained from the different sample types. The workflow presented herein requires low amounts of DNA samples (170 ng) and is cost-effective due to the elimination of DNA and PCR product normalization steps.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.17
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• pp.7641-7651
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• 2015

It has been shown previously that nutritional supplements rich in polyphenolic compounds play a significant role in prostate cancer chemoprevention. Propolis is a natural, resinous hive product that has several pharmacological activities including antimicrobial, antioxidant, anti-inflammatory, and antitumoral activities. The aim of this study was to compare the cytotoxic, antioxidant and antitumoral activities of an ethanolic extract of Egyptian propolis (EEP) in vitro with an established chemotherapeutic drug such as doxorubicin (DOX), and the effects of their combination against the PC3 human prostate cancer cell line. Cellular viability and $IC_{50}$ levels with EEP, DOX and their (v/v) combination were detected by sulphorhodamine-B (SRB) assay after incubation of PC3 cells for 72h with different doses (0, 0.01, 0.1, 1, 10 and $100{\mu}g/ml$). Two selected doses of $IC_{50}$ and $IC_{25}$ were applied to cells for 24h for antitumor evaluation assay of treatment compounds. EEP and its (v/v) combination with DOX showed significant antitumor potential besides high antioxidant properties of superoxide dismutase (SOD), total antioxidant capacity (TAC), catalase (CAT), nitric oxide (NO) and reduced glutathione (GSH) levels when compared with the control untreated cells. DNA fragmentation assay and semi quantitative RT-PCR analyses for p53 and Bax genes showed that EEP activated cellular apoptosis and increased the mRNA expression levels more than other treatment. In conclusion, EEP alone or in combination with DOX at both doses used here showed greater antioxidant, antiproliferative and apoptotic effects against the PC3 cell lines as compared to treatment with DOX alone. Therefore, EEP could be considered as a promising candidate for prostate cancer chemotherapy.

• Journal of Dairy Science and Biotechnology
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• v.33 no.1
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• pp.83-91
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• 2015

Yogurt is a product of the acidic fermentation of milk, which affects the survival of lactic acid bacteria (LAB). The aim of this present study was to examine the survival and acid stress response of Lactobacillus rhamnosus GG to low pH environment. The survival of LAB in commercial yogurt was measured during long-term storage. The enumeration of viable cells of LAB was determined at 15-day intervals over 52-weeks at $5^{\circ}C$. L. acidophilus, L. casei, and Bifidobacterium spp. showed low viability. However, L. rhamnosus GG exhibited excellent survival throughout the refrigerated storage period. At the end of 52-weeks, L. rhamnosus GG survived 7.0 log10 CFU/mL. $F_0F_1$ ATPase activity in L. rhamnosus GG at pH 4.5 was also evaluated. The ATPase activities of the membranes were higher when exposed at pH 4.5 for 24 h. The survival of L. rhamnosus GG was attributable to the induction in $F_0F_1$ ATPase activity. In addition, the mRNA expression levels of acid stress-inducible genes at low pH were investigated by qRT-PCR. clpC and clpE genes were up-regulated after 1 h, and atpA and dnaK genes were up-regulated after 24 h of incubation at pH 4.5. These genes could enhance the survival of L. rhamnosus GG in the acidic condition. Thus, the modulation of the enzymes or genes to assist the viability of LAB in the low pH environment is thought to be important.

• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.2
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• pp.155-162
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• 2002

Cytochrome P45O (CYP) gene has been known to play one of the most important roles in metabolizing the exogenous materials. In insect, CYP is particularly known to detoxify toxic materials by adding oxygen molecule to the hydrophobic region of the materials. Thus, CYP-dependent metabolism is associated with the adaptation of insect to host plant chemicals. This in turn is known to be one of the driving forces for CYP diversification. In the present study, we cloned seven gene fragments of CYP 4 (CYP4) family from the midgut of the beet armyworm, Spodoptera exigua, through RT.PCT, Sequence analysis of the product showed the gene fragment to contain an open reading frame of ~150 amino acids, consisted of ~450 bp. The cloned gene fragments contained typical, conserved regions found in CYP4 family. Pairwise comparison of the deduced amino acid sequences among seven clones ranged in divergence from 0% to 52.86% and resulted in five distinct clones. The other two clones were identical or differ by one amino acid respectively to the corresponding clone, although each differed by ten nucleotides. Analysis of correlation between GenBank-registered, full length CYP4 and the cloned fragments resulted in statistically significant relationship ($r^{2}$ = 0.96085; p < 0.001), suggesting utility of the partial sequences as such full-length sequences. Phylogenetic analysis of the clones with GenBank-registered insect and mammal CYP4 family sequences by parsimony and several distance methods subdivided the clones into two groups: tones belonging to CYP4S and the others to CYP4M families.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.272-278
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• 2010

In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human $y^{+}$LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $y^{+}$LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat $y^{+}$LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest $y^{+}$LAT1 mRNA abundance and the lung had the lowest $y^{+}$LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine $y^{+}$LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.