• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.187-194
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• 2022

Two α-ʟ-arabinofuranosidases (BfdABF1 and BfdABF3) and a β-ᴅ-xylosidase (BfdXYL2) genes were cloned from Bifidobacterium dentium ATCC 27679, and functionally expressed in E. coli BL21(DE3). BfdABF1 showed the highest activity in 50 mM sodium acetate buffer at pH 5.0 and 25℃. This exo-enzyme could hydrolyze p-nitrophenyl arabinofuranoside, arabino-oligosaccharides (AOS), arabinoxylo-oligosaccharides (AXOS) such as 3²-α-ʟ-arabinofuranosyl-xylobiose (A³X), and 2³-α-ʟ-arabinofuranosyl-xylotriose (A²XX), whereas hardly hydrolyzed polymeric substrates such as debranched arabinan and arabinoxylans. BfdABF1 is a typical exo-ABF with the higher specific activity on the oligomeric substrates than the polymers. It prefers to α-(1,2)-ʟ-arabinofuranosidic linkages compared to α-(1,3)-linkages. Especially, BfdABF1 could slowly hydrolyze 2³,3³-di-α-ʟ-arabinofuranosyl-xylotriose (A²⁺³XX). Meanwhile, BfdABF3 showed the highest activity in sodium acetate at pH 6.0 and 50℃, and it has the exclusively high activities on AXOS such as A³X and A²XX. BfdABF3 mainly catalyzes the removal of ʟ-arabinose side chains from various AXOS. BfdXYL2 exhibited the highest activity in sodium citrate at pH 5.0 and 55℃, and it specifically hydrolyzed p-nitrophenyl xylopyranoside and xylo-oligosaccharides (XOS). Also, BfdXYL2 could slowly hydrolyze AOS and AXOS such as A³X. Based on the detailed hydrolytic modes of action of three exo-hydrolases (BfdABF1, BfdABF3, and BfdXYL2) from Bf. dentium, their probable roles in the hemiceullose-utilization system of Bf. dentium are proposed in the present study. These intracellular exo-hydrolases can synergistically produce ʟ-arabinose and ᴅ-xylose from various AOS, XOS, and AXOS.

• Fisheries and Aquatic Sciences
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• v.26 no.3
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• pp.204-215
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• 2023

The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10^-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

• Nutrition Research and Practice
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• v.18 no.1
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• pp.19-32
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• 2024

BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D₃ (1,25[OH]₂D₃) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlr^tm1Her/J (Ldlr^-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)₂D₃ for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)₂D₃ treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)₂D₃ treatment. The 1,25(OH)₂D₃ treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

• IMMUNE NETWORK
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• v.20 no.5
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• pp.40.1-40.15
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• 2020

The protein encoded by the Gene Associated with Retinoid-Interferon-Induced Mortality-19 (GRIM-19) is located in the mitochondrial inner membrane and is homologous to the NADH dehydrogenase 1-alpha subcomplex subunit 13 of the electron transport chain. Multiple sclerosis (MS) is a demyelinating disease that damages the brain and spinal cord. Although both the cause and mechanism of MS progression remain unclear, it is accepted that an immune disorder is involved. We explored whether GRIM-19 ameliorated MS by increasing the levels of inflammatory cytokines and immune cells; we used a mouse model of experimental autoimmune encephalomyelitis (EAE) to this end. Six-to-eight-week-old male C57BL/6, IFNγ-knockout (KO), and GRIM-19 transgenic mice were used; EAE was induced in all strains. A GRIM-19 overexpression vector (GRIM19 OVN) was electrophoretically injected intravenously. The levels of Th1 and Th17 cells were measured via flow cytometry, immunofluorescence, and immunohistochemical analysis. IL-17A and IFNγ expression levels were assessed via ELISA and quantitative PCR. IL-17A expression decreased and IFNγ expression increased in EAE mice that received injections of the GRIM-19 OVN. GRIM19 transgenic mice expressed more IFNγ than did wild-type mice; this inhibited EAE development. However, the effect of GRIM-19 overexpression on the EAE of IFNγ-KO mice did not differ from that of the empty vector. GRIM-19 expression was therapeutic for EAE mice, elevating the IFNγ level. GRIM-19 regulated the Th17/Treg cell balance.

• Microbiology and Biotechnology Letters
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• v.29 no.3
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• pp.181-185
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• 2001

The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.

• Korean Journal of Environmental Biology
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• v.35 no.3
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• pp.389-397
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• 2017

The purpose of this study was to investigate the characteristics of species by comparing the external shape traits and skull size of Tscherskia triton in Jeju Island from May 2014 to November 2016. Body weight, head-body length, ear length, and hind-foot length were not statistically significant (p>0.05), but tail length was statistically significant in females compared to males (p=0.003). To examine the secondary sexual dimorphism characteristics of the collected T. triton, B_HR, T_HR, E_HR, and H_HR were calculated based on the head-body length, and only T_HR was statistically significant (p=0.003). Because of comparing male and female averages, among the 21 traits, the greatest length of skull (GLS; p=0.020) and occipitonasal length (ONL; p=0.014) were statistically significant and length of incisive foramen (LIF) was statistically significant, when the significance level was 90%. However, the other 18 traits did not show statistically significant differences between males and females (p>0.05). Other than GLS, ONL, interorbital breadth (IB), and breadth of occipital foramen (BOF), there was female-biased SSD. The results of this study are the analysis of external morphology and skull characteristics of T. triton in Jeju Island and will be used as basic data to understand the morphological characteristics of T. triton. In addition, the data can be used as a basic information to compare characteristics of T. triton, which is known as a single species, inhabited in China and Russia. The data will play an important role in biogeography and ecology research of T. triton, based on geographical conditions, climate effects, and habitat environment.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.7
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• pp.255-261
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• 2020

This study was undertaken to evaluate the growth, body shape and carcass cutting yield traits of Duroc (D) and crossbred (Duroc×Pietrain×Pietrain; DPP) pigs. A total of 147 D and 101 DPP pigs were used for analyzing the growth trait, whereas 16 D and 16 DPP pigs were evaluated for carcass yields. Backfat thickness (BF) and average daily feed intake (ADFI) were significantly higher in D (14.07±0.24 mm, 2,101 g) than in DPP (12.69±3.25 mm, 1,909 g) (p <0.001). Moreover, D exhibited significantly higher body shape traits including body height (BH), chest depth (CD) and chest width (CW), as compared to DPP pigs (p <0.001). No differences were observed for body length (BL) between the two strains. Analysis of the carcass cutting yield traits determined for D and DPP were in the order: ham (HM; 31.17% and 33.43%), belly (BY; 23.40% and 19.55%), and picnic shoulder (PS; 16.54% and 16.87%), respectively. Then, HM showed a difference of 2.26% P with D(31.17%) and DPP(33.43%), while BY showed a difference of 3.85% P with D(23.40%) and DPP(19.55%). Taken together, our results indicate that DPP has a better feed efficiency than D, and therefore has the potential to increase the production of low-fat pork, targeting consumers having a high preference who have opted for a healthy lifestyle. These results can be used as basic data for developing an ideal pig breed.

• Journal of Life Science
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• v.21 no.8
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• pp.1163-1167
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• 2011

Excessive alcohol consumption causes various degenerative brain diseases including Alzheimer's disease and Parkinson's disease. Absorbed ethanol is metabolized to acetaldehyde and acetic acid by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Acetaldehyde is well known as a toxicant through generation of reactive oxygen species (ROS). Therefore, ALDH2 activity may play important roles in the pathogenesis of alcohol-induced brain diseases. In this study, we demonstrated the effects of ALDH2 enzyme activity on lipid peroxidation in brain tissues and urine of mice exposed to ethanol for 8 weeks. Five male, 8-week old Aldh2 (+/+) and Aldh2 (-/-) mice (C57BL/6J strain) in each group were exposed to ethanol for 8 weeks (2 g/kg wt./day) using gavage, and those in the control group received 0.9% saline alone. Thiobarbituric acid reactive substances (TBARS) level, a marker for lipid peroxidation, was measured in whole brain tissue and urine by high performance liquid chromatography. As a result, chronic ethanol treatment did not show any statistical change on the TBARS level of brain tissue in both Aldh2 (+/+) mice and in Aldh2 (-/-) mice. However, following ethanol exposure for 8 weeks in Aldh2 (-/-) mice, the urinary TBARS levels were significantly increased to more than double compared to the pretreatment group. This result was not observed in Aldh2 (+/+) mice. These results suggest that although ALDH2 enzyme activity plays a role in the generation of ROS in the whole body, it does not seem to be important in the pathogenesis of alcohol induced degenerative brain diseases.

• Journal of Oriental Neuropsychiatry
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• v.21 no.4
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• pp.21-39
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• 2010

Objectives : This study was to investigate the characteristics of the heart rate variability(HRV), types of insomnia and accompanying symptoms of 175 insomnia patients. Methods : For this study we carried out HRV, Inbody of 175 patients with insomnia who have come to Dongeui oriental hospital of Dongeui university from the period of Janaury 2008 to September 2010. We studied the association of the insomnia patients' age and gender with HRV, accompanying symptoms and the types of insomnia. The information of the accompanying symptoms and types of insomnia was based on each patients' progress note. Results : l. There was no significant differences in the characteristics of HRV between male and female. 2. The most frequent symptom shown among the insmonia patents' was headache(41.14%). 3. Comparing the symptoms between the gender, female patients had more dry mouth, alternative periodic chill and fever, and hot flush symptoms than the male patients. 4. Comparing the symptoms between the age groups(divided in two), the lower age group (20~59) had more dizziness and constipation symptoms than the higher age group (over 60). 5. Comparing the types of insomma between the age group, the higher age group (over 60) had more termination insomnia. 6. The types of insomnia of the entire insomnia patients were onset insomnia (73.1%), maintenance insomnia(20.6%), termination insomma(l7.1 %), shallow sleep (52.6%), listed by order of frequency. 7. The patients who takes hypnotics had more anorexia symptoms than the patients who doesn't take hypnotics. 8. There was no visible differences of the average fractal portrait between male and female. The age group of 20~30yrs. had the highest average fractal portrait, and the age group of 30~59yrs. the second highest, and the age group over 60yrs. the lowest. 9. The average of the regulation reserves of the autonomic nervous system(B2) - was lower than the regulation level of the ANS at the present (Bl). 10. When compared the priority of the function of the ANS, it showed that the proportion of HF($38.61{\pm}29.19%$)was the most, and than VLF($30.65{\pm}23.36%$), LF($20.04{\pm}19.13%$) the least. 11. The average of the compensation level of the central nervous system at present(Cl) - was lower than the compensation reserves of the CNS(C2). 12. The average of the control reserves of the cerebrum(D2) - was lower than the control level of the cerebrum at the present (Dl). 13. There was no visible differences between different sexes and ages in pulse rate. 14. The abdomen fat ratio above the line of the insomnia patients was 77.97% in male and 93.1% in female. Both sexes showed that insomnia patients had more abdomen fat that the standard, and female patients had more abd. fat than male patients. Conclusions : This study shows that the HRV of insomnia patients had no significant differences between gender. Fractal portrait, HF, LF and VLF of the insomnia patients are in inverse proportion to the age. The study of the Body Component Analysis showed that female had more abdomen fat than male, and both gender showed more abdomen fat than the standard. When looked into the accompanying symptoms of the insomnia patients, the symptoms show differences according to gender, age and hypnotics taking, as shown as below. In the entire patients, Headache was the most accompanying symptom. Female had more dry mouth, alternative periodic chill and fever, and hot flush symptoms than male. Higher age groups had more dizziness and constipation as accompanying symptoms than lower age groups. Patients who takes hypnotics had more anorexia than those who dont.

• Journal of Gastric Cancer
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• v.6 no.1
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• pp.6-10
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• 2006

Purpose: The most common metabolic defect appearing following a gastrectomy is anemia. Two types have been identified: One is related to a deficiency in iron and the other is related to an impairment in vitamin B12 metabolism. The purpose of this study is to evaluate the incidence and treatment of vitamin B12 deficiency after a total gastrectomy. Materials and Methods: Serum vitamin B12 concentrations were measured in 128 patients who had undergone a total gastrectomy. The group with a serum concentration under 200 pg/ml was supplemented at one-month intervals with Actinamide and five-six months later, serum concentrations of the vitamin B12 were rechecked. Results: The group with vitamin B12 under 200 pg/ml was 61 (47.6%) of the 128 patients who had undergone a total gastrectomy. In this group, the cumulative incidences of vitamin B12 deficiency were 7.0, 23.4, 33.6, 39.1, 41.4, and 47.7% at 6 months, 1, 2, 3, 4, and 5 or more years, respectively. The treated group with vitamin B12 under 200 pg/ml had 36 (28.17%) of the 128 patients. The 16 of those cases with vitamin B12 levels of $100{\sim}150\;pg/ml$ were supplemented 5.21 times and the vitamine B12 was elevated to above 650 pg/ml. The other 20 cases with an average of vitamin B12 levels of $150{\sim}200pg/ml$ were supplemented an average of 4.75 times, and the vitamin B12 was elevated to above 780 pg/ml. Conclusion: It is necessary to supplement vitamin B12 even 1 year later after a total gastrectomy. The group with vitamin B12 under 200 pg/ml was supplemented $5{\sim}6$ times at one-month intervals with Actinamide $1,000\;{\mu}g$ IM injections and reached normal levels. (J Korean Gastric Cancer Assoc 2006;1:6-10)