• Journal of the Korean Wood Science and Technology
• /
• v.46 no.6
• /
• pp.670-680
• /
• 2018

Acanthophysium sp. KMF001, a wood rotting fungus, produces a strong crude enzyme complex that efficiently produces simple sugars from wood. The transcriptomic analysis of Acanthophysium sp. KMF001 identified 14 genes for putative glycoside hydrolases. Among them, isotig01043 was expressed heterogeneously in Escherichia coli BL21(DE3), and the expressed protein exhibited an endo-${\beta}$-1,4-xylanase activity which showed the optimum reaction at pH 5.0 and $30^{\circ}C$. The enzyme kinetic values of $K_m$ and $V_{max}$ were 25.92 mg/ml and $0.628{\mu}mole/mg/ml$, respectively. The enzymatic characteristics of the expressed xylanase showed a typical fungal xylanase. However, the bioinformatics analysis suggested that the protein encoded by isotig01043 was a novel xylanase based on a low identity when it was compared with the closest protein in the NCBI database and a similar protein domain with GH16_fungal_Lam16A_glucanase, which had not been earlier suggested as a xylanase.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.15
• /
• pp.6663-6668
• /
• 2015

Background: Treatment failure in leukemia is due to either pharmacokinetic resistance or cell resistance to drugs. Materials and Methods: Gene expression of multiple drug resistance protein (MDR-1), multidrug resistance-related protein (MRP) and low resistance protein (LRP) was assessed in 45 pediatric ALL cases and 7 healthy controls by real time PCR. The expression was scored as negative, weak, moderate and strong. Results: The male female ratio of cases was 2.75:1 and the mean age was 5.2 years. Some 26/45 (58%) were in standard risk, 17/45(38%) intermediate and 2/45 (4%) in high risk categorie, 42/45 (93%) being B-ALL and recurrent translocations being noted in 5/45 (11.0%). Rapid early response (RER) at day 14 was seen in 37/45 (82.3%) and slow early response (SER) in 8/45 (17.7%) cases. Positive expression of MDR-1, LRP and MRP was noted in 14/45 (31%), 15/45 (33%) and 27/45 (60%) cases and strong expression in 3/14 (21%), 11/27 (40.7%) and 8/15 (53.3%) cases respectively. Dual or more gene positivity was noted in 17/45 (38%) cases. 46.5 % (7/15) of LRP positive cases at day 14 were in RER as compared to 100% (30/30) of LRP negative cases (p<0.05). All 8 (100%) LRP positive cases in SER had strong LRP expression (p=<0.05). Moreover, only 53.3% of LRP positive cases were in haematological remission at day 30 as compared to 100% of LRP negative cases (p=<0.05). Conclusions: Our study indicated that increased LRP expression at diagnosis in pediatric ALL predicts poor response to early treatment and hence can be used as a prognostic marker. However, larger prospective studies with longer follow up are needed, to understand the clinical relevance of drug resistance proteins.

• KSBB Journal
• /
• v.28 no.2
• /
• pp.137-145
• /
• 2013

In order to develop new skin whitening agents, we prepared the $CH_2Cl_2$ layer (NGC) and BuOH layer (NGB) of 75% EtOH extract of the Nelumbinis nucifera Gaertner. We measured their tyrosinase inhibitory activity in vitro and melanin synthesis inhibitory activity in B16-F1 melanoma cells. They did not show inhibitory activity against mushroom tyrosinase but showed melanin synthesis inhibitory activity in a dose-dependent manner. In a melanin synthesis inhibition assay, NGC and NGB suppressed melanin production up to 52% and 46% at a concentration of $100{\mu}g/mL$, respectively. To elucidate the mechanism of the inhibitory effects of NGC and NGB on melanogenesis, we measured the expression of melanogenesis-related proteins by western blot assay. As a result, NGC suppressed the expression of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), phosphorylated cAMP responsive element binding (p-CREB) protein, and microphthalmia associated transcription factor (MITF). And NGB inhibited the protein expression of tyrosinase and MITF, but had no significant effect on TRP-1, TRP-2, and p-CREB expression. Moreover, NGB increased the expression of phosphorylated extracellular signal-regulated kinase (p-ERK). In addition, we examined the inhibitory effect on the glycosylation of tyrosinase. As a result, NGC and NGB inhibited the activity of ${\alpha}$-glucosidase in vitro and the glycosylation of tyrosinase in B16-F1 melanoma cells. From these results, we concluded that NGC and NGB could be used as active ingredients for skin whitening.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.16 no.2
• /
• pp.87-92
• /
• 2008

Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

• Journal of Yeungnam Medical Science
• /
• v.16 no.1
• /
• pp.43-51
• /
• 1999

The aim of present study was to elucidate whether the expression of nm23 protein might be of clinical value as a prognostic factor in gastric cancer. The expression of nm23 protein was analyzed using an immunohistochemical method with formalin-fixed and paraffin embedded tissue samples from 76 gastric carcinoma patients. The cytoplasmic immunoreactivity of nm23 protein were detected in 53.9% of the sample tissues(41/76). When the immunoreactivity of nm23 protein with TNM status and other histopathologic findings were compared by using Chi-Square test, nm23 was found to have correlations with lymph node metastasis(p=0.04), a number of metastatic lymph node, and the invasion of lymphatic vessels(p=0.007); however, it had no correlation with TNM status. The conventional prognostic factors such as the depth of invasion, the degree of lymph node metastasis and the presence of distant metastasis, a Borrmann type, size of tumor, and the curability with operation were found to have a strong correlation with the survival time(p<0.003). However, the expression of nm23 protein was not significantly correlated with survival time in survival analysis. These results showed that the expression of nm23 protein is not a useful prognostic indicator in gastric cancer.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.5
• /
• pp.598-606
• /
• 2010

This study investigated the nutritional quality of soybean meal (SBM) fermented by Aspergillus ($FSBM_A$) and/or followed by Lactobacillus fermentation ($FSBM_{A+L}$). Both fermented products significantly improved protein utilization of SBM with higher trichloroacetic acid (TCA) soluble true protein content, in vitro protein digestibility and available lysine content, especially in $FSBM_{A+L}$. Moreover, $FSBM_{A+L}$ produced a huge amount of lactic acid resulting in lower pH as compared to the unfermented SBM or soybean protein concentrate (SPC) (p<0.05). $FSBM_A$ and $FSBM_{A+L}$ raised 4.14% and 9.04% of essential amino acids and 5.38% and 9.37% of non-essential amino acids content, respectively. The ${\alpha}$-galactoside linkage oligosaccharides such as raffinose and stachyose content in $FSBM_A$ and $FSBM_{A+L}$ decreased significantly. The results of soluble protein fractions and distribution showed that the ratio of small protein fractions (<16 kDa) were 42.6% and 63.5% for $FSBM_A$ and $FSBM_{A+L}$, respectively, as compared to 7.2% for SBM, where the ratio of large size fractions (>55 kDa, mainly ${\beta}$-conglycinin) decreased to 9.4%, 5.4% and increased to 38.8%, respectively. There were no significant differences in ileal protein digestibility regardless of treatment groups. SPC inclusion in the diet showed a better protein digestibility than the SBM diet. In summary, soybean meal fermented by Aspergillus, especially through the consequent Lactobacillus fermentation, could increase the nutritional value as compared with unfermented SBM and is compatible with SPC.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.10
• /
• pp.1443-1450
• /
• 2003

Crossbred (Bos taurus${\times}$Bos indicus) calves were used from birth till 14 weeks of age to evaluate three sources of protein that differed in ruminal degradability viz. groundnut cake alone (HD) or in combination with cottonseed meal (MD) and meat and bone meal (LD), when fed along with two sources of non-structural carbohydrates viz. raw (R) and thermally processed (P) maize. Twenty four new born calves were arranged in six groups in a $3{\times}2$ factorial design and fed on whole milk up to 56 d of age. All the different calves received calf startes along with green oats (Avena sativa) from 14 d of age onwards free-choice. A metabolism trial of 6d starters duration, conducted after 90 d of experimental feeding, revealed greater (p<0.05) digestibility of DM, OM, total carbohydrates, NDF and ADF in calves fed on the P diets than on the R diets promoting greater (p<0.05) metabolizable energy intake. The digestibility of NDF was higher (p<0.01) on LD diets where as calves on MD diets exhibited significantly lower digestibility of ADF (p<0.01). The retention of nitrogen per unit metabolic body size was significantly (p<0.05) higher on the LD-P diet than on the diet HD-P which, in turn, was higher (p<0.05) than that of HD-R. Nitrogen retention as percentage of intake was significantly greater (p<0.05) on LD-P than on LD-R diets (52.2 vs. 36.4%). Also, P fed calves utilized nitrogen more efficiently than the R fed as shown by retention of significantly greater proportions of intake (47.4 vs. 40.9%) and absorbed (65.8 vs. 59.5%) nitrogen. Calorimetric evaluation of the diets through open-circuit respiration chamber revealed that the dietary treatments had no impact on methane production by calves. The intake of DE and ME was improved (p<0.01) because of maize processing resulting in greater (p<0.01) retention of energy. The protein degradability exerted no influence on the partitioning or retention of energy. A significant interaction between cereal and protein types was evident with respect to retention of both nitrogen (p<0.01) and energy (p<0.05). In conclusion, no discernible trend in the influence of cereal processing was apparent on the dietary protein degradability, but the positive effect of cereal processing on energy retention diminished with the increase in dietary undegradability.

• Tuberculosis and Respiratory Diseases
• /
• v.73 no.1
• /
• pp.11-21
• /
• 2012

Background: While qualitative analysis of methylation has been reviewed, the quantitative analysis of methylation has rarely been studied. We evaluated the methylation status of CDKN2A, $RAR{\beta}$, and RASSF1A promoter regions in non-small cell lung carcinomas (NSCLCs) by using pyrosequencing. Then, we evaluated the association between methylation at the promoter regions of these tumor suppressor genes and the clinicopathological parameters of the NSCLCs. Methods: We collected tumor tissues from a total of 53 patients with NSCLCs and analyzed the methylation level of the CDKN2A, $RAR{\beta}$, and RASSF1A promoter regions by using pyrosequencing. In addition, we investigated the correlation between the hypermethylation of CDKN2A and the loss of $p16^{INK4A}$ immunoexpression. Results: Hypermethylation of CDKN2A, $RAR{\beta}$, and RASSF1A promoter regions were 16 (30.2%), 22 (41.5%), and 21 tumors (39.6%), respectively. The incidence of hypermethylation at the CDKN2A promoter in the tumors was higher in undifferentiated large cell carcinomas than in other subtypes (p=0.002). Hyperrmethylation of CDKN2A was significantly associated with $p16^{INK4A}$ immunoexpression loss (p=0.045). With regard to the clinicopathological characteristics of NSCLC, certain histopathological subtypes were found to be strongly associated with the loss of $p16^{INK4A}$ immunoexpression (p=0.016). Squamous cell carcinoma and undifferentiated large cell carcinoma showed $p16^{INK4A}$ immunoexpression loss more frequently. The Kaplan-Meier survival curves analysis showed that methylation level and patient survival were barely related to one another. Conclusion: We quantitatively analyzed the promoter methylation status by using pyrosequencing. We showed a significant correlation between CDKN2A hypermethylation and $p16^{INK4A}$ immunoexpression loss.

• The Korean Journal of Physiology and Pharmacology
• /
• v.16 no.3
• /
• pp.199-204
• /
• 2012

We evaluated the role of Tat-mediated p66shc transduction on the activation of endothelial nitric oxide synthase in cultured mouse endothelial cells. To construct the Tat-p66shc fusion protein, human full length p66shc cDNA was fused with the Tat-protein transduction domain. Transduction of TAT-p66shc showed a concentration- and time-dependent manner in endothelial cells. Tat-mediated p66shc transduction showed increased hydrogen peroxide and superoxide production, compared with Tat-p66shc (S/A), serine 36 residue mutant of p66shc. Tat-mediated p66shc transduction decreased endothelial nitric oxide synthase phosphorylation in endothelial cells. Furthermore, Tat-mediated p66shc transduction augmented TNF-${\alpha}$-induced p38 MAPK phosphorylation in endothelial cells. These results suggest that Tat-mediated p66shc transduction efficiently inhibited endothelial nitric oxide synthase phosphorylation in endothelial cells.

• Journal of the East Asian Society of Dietary Life
• /
• v.9 no.2
• /
• pp.169-176
• /
• 1999

The purpose of this study was to investigate the relationships of dietary nutrients and serum heavy metals in college women living Choong-Nam area of Korea. The mean age, height, weight, and BMI were 22.9years, 158.74cm, 53.39kg, and 21.71kg/$m^2$ respectively. The mean daily energy intake was 85.9% of RDA for Koreans. The ratio of energy from carbohydrate, fat, and protein was 61:23:16. And the daily vitamin A, B$_2$, Ca were 90%, 78%, 60% of RDA for Korea, respectively. The mean serum levels of Pb, Cd, Cr were 0.190, 0.005, 0.025$\mu\textrm{g}$/$m\ell$, respectively. The serum Cd was significantly different with dietary carbohydrate(p＜0.05). And the serum Cr was significantly different with dietary protein intake(p＜0.05), phosphorus(p＜0.01), potassium(p＜0.05). respectively.