• The Journal of Korean Medicine
• /
• v.32 no.3
• /
• pp.25-34
• /
• 2011

Objectives: We performed simultaneous determination of five constituents by HPLC in Samul-tang (SMT). Additionally, we investigated the immune-stimulatory potential of SMT on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 190-400 nm, were used for quantification of the five components of SMT. Mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. The extract of SMT (1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Results: Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD, %) for intra- and inter-day precision were both less than 3.5%. The recovery was in the range of 95.69-115.12%, with an RSD less than 6.0%. The contents of five components in SMT were 1.08-15.30 mg/g. SMT significantly enhanced Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). Also, SMT significantly enhanced OVAspecific IgG, IgG1 and total IgM levels in plasma compared with the OVA-immunized group. Conclusions: The established method will be applied for the quantification of major components and immunestimulating activity in OVA-immunized mouse model of SMT.

• Advances in nano research
• /
• v.4 no.3
• /
• pp.167-179
• /
• 2016

The detailed study of acute and sub-acute toxicity of the complex polyvinylpyrrolidon (PVP 20 kDa)-wrapped fullerene $C_{60}$ after intraperitoneal (ip) administration was carried out on adult male Wistar rats. The $LD_{50}$ value of $C_{60}/PVP$ complex was found to be 7, 8 g/kg. In sub-acute study which lasted for 30 days the rats were exposed to daily administration of the complex in the doses of 350 or 700 mg/kg. All animals survived during the study and had no significant changes in clinical signs, organ weight, hematological and biochemical parameters of blood. The electrophysiological properties of myocardium and the excretory function of kidneys remained normal. Histological analysis of liver, kidney and spleen at the end of the study also did not demonstrate toxic alterations. It was thus established that intraperitoneal administration of complex $C_{60}/PVP$ has no toxic effect. These results suggest that $C_{60}/PVP$ has no acute and sub-acute toxicity and is a perspective substance for potential application in biology and medicine.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.1
• /
• pp.9-18
• /
• 2017

Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity ($83.23mU/{\mu}l$) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The $K_m$ and $V_{max}$ values of recombinant AprE34 were $0.131{\pm}0.026mM$ and $16.551{\pm}0.316{\mu}M/l/min$, respectively, when measured using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.7
• /
• pp.1108-1114
• /
• 2007

Twenty eight non-lactating and non-pregnant adult Serra da Estrela ewes, ranging in body condition score (BCS) from 1 to 4 were used to study the relationships between BCS, live weight (LW), body composition and fat partition. Ewes were slaughtered and their kidney knob and channel fat (KKCF), sternal fat (STF) and omental plus mesenteric fat (OMF) were separated and weighed. Left sides of carcasses as well as the respective lumbar joints were then dissected into muscle, bone and subcutaneous (SCF) and intermuscular fat (IMF). The relationship between LW and BCS was studied using data from 1,396 observations on 63 ewes from the same flock and it was found to be linear. Regression analysis was also used to describe the relationships among BCS and/or LW and weights (kg) and percentages in empty body weight (EBW) of dissected tissues. The prediction of weights and percentages in EBW of total fat (TF) and of all fat depots afforded by BCS was better than that provided by LW. Only the weight of muscle and the percentage of bone in the EBW were more efficiently predicted by LW than by BCS. IMF represented the largest fat depot with a BCS of 1 and 2, whereas SCF was the most important site of fat deposition with a BCS of 3 and 4. Allometric coefficients for each fat depot in TF suggest that the fat deposition order in ewes from this breed is: IMF, OMF, SCF and KKCF. Results demonstrate that BCS is a better predictor than LW of body reserves in this breed and that LJ is a suitable anatomical region to evaluate BCS.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.3
• /
• pp.474-482
• /
• 2004

The $\alpha$-L-arabinofuranosidase (Arfase) gene of Bacillus stearothermophilus No. 236 was cloned and sequenced. The ORF of the gene, designated arfA, encoded a 507 -residue polypeptide with calculated molecular mass of 57 kDa. The Arfase produced by a recombinant Escherichia coli strain containing the arfA gene was purified to apparent homogeneity and characterized. The molecular mass of the Arfase determined by SDS-PAGE was 60 kDa. However, according to gel filtration, it was estimated to be approximately 190 kDa. These results indicated that the functional form of the Arfase is trimeric. The optimal pH and temperature for the enzyme activity were pH 6.5 and $55^{\circ}C$, respectively. The half-life of the enzyme at $60^{\circ}C$ was about 6 h. Kinetic experiments at $45^{\circ}C$ with pNPM (p-nitrophenyl $\alpha$-L-arabinofuranoside) as a substrate gave the $K_m and V_{max}$ values of 1.19 mM and 26.1 U/ mg, respectively. When the enzyme was combined with Bacillus stearothermophilus No. 236 endoxylanase and $\beta$-xylosidase, it hydrolyzed arabinoxylan into L-arabinose and xylose more efficiently than Arfase alone. This synergistic effect suggested that the complete hydrolysis of xylan with large amounts of arabinose side chains required Arfase as well as endoxylanase and $\beta$-xylosidase.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.148.2-149
• /
• 2003

Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

• Korean Journal of Veterinary Service
• /
• v.43 no.2
• /
• pp.67-77
• /
• 2020

Canine infectious respiratory disease (CIRD), also known as infectious tracheobronchitis or kennel cough occurs in a multiple-dog environment such as a shelter. In this study, we were collected 300 of nasal swab samples from dogs and 145 of environmental samples from a shelter to investigate respiratory pathogens of dogs in the Gwangju metropolitan city animal shelter from February to October, 2019. Bacteria cultures for isolation of Bordetella (B.) bronchiseptica and polymerase chain reaction (PCR) tests were performed for detection of eleven canine respiratory pathogens, namely Mycoplasma (M.) cynos, canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine respiratory coronavirus (CRCoV), alpha-coronavirus (CCoV), canine pneumovirus (CnPnV), canine hepacivirus (CHeV), canine adenovirus type 2 (CAdV-2), canine herpesvirus-1 (CHV-1) and canine bocavirus (CBoV). Among 300 nasal swab samples, 148 samples (49.3%) were positive for at least one pathogens. CHV-1 was the most common pathogen, found in 95/300 (31.7%) samples. Subsequently, M. cynos (22.0%), B. bronchiseptica (2.3%), CPIV (2.0%), CBoV (1.7%), CCoV (0.7%) were detected. The detection rates of M. cynos and CHV-1 according to the duration of stay in the shelter were statistically significant. Among environmental samples, M. cynos, CCoV, CBoV and CHV-1 were detected in 45/145 (31.0%). These results indicated the need for disease control and prevention systems in the shelter.

• Korean Journal of Microbiology
• /
• v.30 no.2
• /
• pp.101-107
• /
• 1992

Cytochrome c oxida5e from chemotrophically grown R p , geliitinosu was purified by cytochrome c affinity chromatography and DEAE-Sephacel ion exchange chromatography. The molecular weight of the cytochrome c oxidase was approximately 110.000 Da by sephacryl s-300 gel chromatography and approximately 52, 000 Da by SDS-gel electrophoresis, respectively. Therefore. cytochrolne c oxidase of Rps. gehtinosu seems to be dimer. The cytochrome c oxidasc was very sensitive to temperature. It's Km and Vmax were 20 pM and 44 unitlmg protein for horsc heart cytochrome c as a substrate. respectively, and its optimum pH and temperature were 6.4 and 25$^{\circ}$C. respectively. The absorption peaks of the reduced cytochrome c oxidase showed at 554 nm, 523 nm. and 422 nm. The activiiy of cytochrome c oxidase was inhibited by KCN, and NaN3, but not by CO, antimycir~ A. and myxothiazol. The cytochrome c-551 was produced either in phototrophically or chemotrophically grown Rps. gelaiinosci. The rcduced cytochrome c-551 was oxidized by b-type cytochrome c oxidase from Rp.v. gc.lrtino.sc~. Km and Vmax of cytochrome c oxidase was 26 pM and 31 unitlnlg protein For cytochrome c-551 as a substrate. respectively. Thercfore. thc electron transfer chain of chemotrophically grown Rps. glatinosa seems lo be ubiquinol cytochrome bc, complex -'cytochrome c-55lMb-type cytochrome c oxidase+02.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.46 no.10
• /
• pp.1164-1170
• /
• 2017

Protaetia brevitarsis larvae (PBL) has recently been registered as a temporary food in Korea, and this study evaluated the application potential of PBL proteins as health functional food materials. Protein hydrolysates were prepared from PBL powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and based on the results from the peptide content and SDS-PAGE analyses, PBL treated with alcalase or flavourzyme showed a high degree of hydrolysis (HD) value, whereas the HD value of those treated with neutrase, bromelain, or papain was minimal. The protein hydrolysates showing a high HD value were separated further into the fractions of >3 kDa and <3 kDa by a centrifugal filter system and then lyophilized, and according to the $RC_{50}$ values of the protein hydrolysates (<3 kDa) obtained from three different antioxidant analyses; the alcalase hydrolysates showed the highest antioxidant activity. Therefore, the alcalase hydrolysates were tested further for their inhibitory effects on the peroxidation of linoleic acid by measuring the thiobarbituric acid values. The results showed that the peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation, but a pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited the linoleic acid peroxidation in a dose-dependent manner for 6 days. Our current studies are focused on the identification of active peptide sequences from alcalase hydrolysates.

• Nuclear Engineering and Technology
• /
• v.55 no.7
• /
• pp.2349-2355
• /
• 2023

In the TRITIUM project, an on-site monitoring system is being developed to measure tritium (³H) levels in water near nuclear power plants. The quite low-energy betas emitted by ³H have a very short average path in water (5 ㎛ as shown by simulations for 18 keV electrons). This path would be further reduced by impurities present in the water, resulting in a significant reduction of the detection efficiency. Therefore, one of the essential requirements of the project is the elimination of these impurities through a filtration process and the removal of salts in solution. This paper describes a water treatment system developed for the project that meets the following requirements: the water produced should be of near-pure water quality according to ISO 3696 grade 3 standard (conductivity < 10 µS/cm); the system should operate autonomously and be remotely monitored.