• Journal of Applied Biological Chemistry
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• v.66
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• pp.424-435
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• 2023

As a result of this study, DPPH radical scavenging activity was 81.8% at a concentration of 100 ㎍/mL of Alpinia galanga L. rhizome 70% ethanol extract (AG.E), and ABTS⁺ radical scavenging activity was confirmed to be 99.8%, similar to L-Ascorbic acid (AA), at a low concentration of 50 ㎍/mL AG.E. To measure anti-aging activity, collagenase and elastase inhibitory activities were measured and AG.E showed higher inhibitory effects than epigallocatechin gallate (EGCG) starting at a low concentration of 50 ㎍/mL. In particular, AG.E showed inhibitory effects more than three times that of EGCG at a concentration of 500 ㎍/mL. In order to verify anti-aging effect of AG.E in CCD-986sk cell, good anti-aging effect was obtained in various experiments stimulated with UVB. In a gene expression analysis experiment using RT-PCR, the COL1A mRNA expression level was found to increase 2.90 times compared to no addition at a low concentration of 20 ㎍/mL AG.E, confirming the possibility of developing it as a good functional material related to anti-aging. As a basic study on temporal biological activity preservation ratio of material when applied to formulations, AG.E, and AA were added to a stable W/O type emulsion and stored in a thermostat at 25 ℃ for 60 days. As a result of measuring DPPH and ABTS⁺ radical scavenging activities on the 1st, 30th, and 60th days, it was confirmed that antioxidant effects are maintained at a high level over time in formulations.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.23 no.2
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• pp.39-44
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• 2023

Hereditary tyrosinemia type 1 (HT-1) is a metabolic disorder caused by biallelic pathogenic variants in the fumarylacetoacetate hydrolase (FAH) gene, which impairs the function of the FAH enzyme, resulting in the accumulation of tyrosine's toxic metabolites in hepatocytes and renal tubular cells. As a consequence, individuals with HT-1 exhibit symptomatic manifestations. Rapid diagnosis and treatment of HT-1 can prevent short-term death and long-term complications. A 15-day-old boy presented to the outpatient department with elevated levels of tyrosine on his newborn screening tests conducted at the age of 3 and 10 days, respectively. Further blood tests revealed increased levels of alpha-fetoprotein and amino acids including tyrosine and threonine. Urine organic acid tests indicated a significant elevation in tyrosine metabolites, as well as the presence of succinylacetone (SA), which led to the diagnosis of HT-1. Two pathogenic and likely pathogenic variants of FAH compatible with HT-1 were also detected. He began a tyrosine-restricted diet at one month old and received nitisinone (NTBC) at two months old. With continued treatment, the patient's initially elevated AFP level, detection of SA in the urine, and mild hepatomegaly showed improvement. During four years and seven months of treatment, there were no exceptional complications apart from an increase in tyrosine levels and a delay in speech. We report a case of tyrosinemia type 1 detected through newborn screening, treated with dietary restriction and NTBC, with a good prognosis.

• Journal of the Korean Chemical Society
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• v.51 no.3
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• pp.251-257
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• 2007

The Tumor necrosis factor-α, (TNF-α), is involved in the pathogenesis of multiple sclerosis and contributes to the degeneration of oligodendrocytes as well as neurons. Nicotine has been found to have immunosuppressive and inflammation-suppressing effects. Astrocytes, the major glial cells in the CNS, are capable of producing TNF-α at both the mRNA and protein levels in response to interleukin-1 (IL-1) or TNF-α. Nicotine has been shown to influence glial cell functions. To order to explore the role of astrocytes in the production of TNF-α, astrocytes were pretreated with nicotine and are stimulated with IL-1β to determine their effects on TNF-α production. The results are as follows. Cytotoxic effects of nicotine on human fetal astrocytes were noted above 10 μg/ml of nicotine. The effect of IL-1β on TNF-α mRNA expression in primary cultured human fetal astrocytes was maximal at 2 h after IL- 1β(100 pg/ml) treatment. Human fetal astrocytes were pretreated with 0.1, 1, and 10 μg/ml of nicotine and then stimulated with IL-1β (100 pg/ml) for 2 h. The inhibitory effect of nicotine on expressions of TNF-α mRNA in human fetal astrocytes with pretreated 0.1 μg/ml of nicotine is first noted at 8 hr, and the inhibitory effect is maximal at 12 h. The inhibitory effect at 1 μg/ml of nicotine is inhibited maximal at 24 h. Nicotine at 0.1, 1 and 10 μg/ml concentrations significantly inhibits IL-1β-induced NF-κB activation. Collectively, this study indicates that nicotine might inhibit the expression of TNF-α in activated human fetal astrocytes.

• Journal of Life Science
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• v.28 no.9
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• pp.1056-1061
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• 2018

This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

• Journal of Life Science
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• v.22 no.6
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• pp.703-712
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• 2012

Adeno-associated virus (AAV) has been considered to be a very safe and efficient gene delivery system. However, the major obstacles to therapeutic usage of AAV have been to achieve highly efficient and reproducible production processes, and also to develop a reliable quantifying method of various serotypes with a simple protocol. We compared the efficiency of the conventional production protocol of AAV2 and adenovirus (Ad) co-infection to that of a new method containing AAV2 infection followed by pHelper transfection. We tested HEK293 and 293T, and further examined the time-dependent changes of AAV2 production. The new method of AAV2 and pHelper DNA gave about ten times higher production efficiency than that of the conventional protocol. The highest production efficiency in 293T was achieved as $1.61{\times}10^5$ virus genomes (v.g.)/cell by the new method of 10 MOI of AAV2 infection and 5 days post-infection. This protocol of the highest efficiency was then applied to produce various AAV serotypes and showed the efficiencies higher than $10^5$ v.g./cell. Next, we designed the universal PCR primers of highly conserved regions for various AAV serotypes to develop a simple and reliable titration method. The universal primers could amplify all the tested AAV serotypes with similar sensitivities by ten molecular copies. Therefore, this pair of universal primers can be further utilized to detect AAV contaminants in therapeutic adenoviral vectors.

• Pediatric Infection and Vaccine
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• v.22 no.2
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• pp.106-112
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• 2015

Purpose: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. Methods: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. Results: The detection limit of the PCR assay was approximately $10^4$ colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over $10^6$ CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). Conclusions: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA -specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.

• Microbiology and Biotechnology Letters
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• v.33 no.1
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• pp.16-23
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• 2005

In order to develop a new microbial fungicide using endophytic bacteria for the control of anthracnoses occurring on various crops, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth medium, their antifungal activities were tested for in vivo antifungal activity against cucumber anthracnose caused by Colletotrichum orbiculare. As the results, liquid cultures of 28 strains showed potent antifungal activities more than $90\%$ against cucumber anthracnose. At 3-fold dilutions of liquid cultures, 18 strains inhibited the development of cucumber anthracnose of more than $70\%$. They were further tested for in vivo antifungal activity against red pepper anthracnose caused by C. coccodes and in vitro antifungal activity against C. acutatum, a fungal agent causing red pepper anthracnose. Among 18 strains, a bacterial strain EB215 isolated from cucumber roots displayed the most potent antifungal activity against Colletotrichum species. It was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, Biolog test and 16S rDNA gene sequence. It also controlled effectively the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), and tomato late blight (Phytophthora infestans). Studies on the characterization of antifungal substances produced by B. cepacia EB215 are in progress.

• Journal of Sericultural and Entomological Science
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• v.46 no.2
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• pp.65-71
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• 2004

Long term preservation experiment through refrigeration was conducted for 2 year on 300 lines of silkworm races preserved, as one method for the development of long term safe preservation technique. Experiment with 6 treatments was conducted for 680 days from July 1 st 2000 to May 1st 2002. Embryonic development was conducted to each treatment. There are no differences among treatments and races in 400 days preservation, the stage of whole embryonic development was Eul B and condition of eggs was good. In 650 days preservation experiment, differences were reveled among races and treatment, the level of whole embryonic development was Byeong A and condition of eggs was good. The order of embryonic development is European races >Tropical, Korean races >Japanese, Chinese races, thus European races showed fast embryonic development. Control(treatment A) and treatment C showed faster development than other treatments. And treatment D and F showed stable individual stage among all treatments. The test of shape characteristics and embryo which were conducted in hatching period showed 61% of high line succession possibility in average of 6 treatments. But treatment A and B showed no hatching, 3 lines of treatment C, 48 lines of treatment D, 1 line of treatment E, and 29 lines of treatment F showed slow development. And treatments D and F which showed stable embryo condition had highest possibility. The two treatments D and F showed good result among six treatments, and the preservation period of treatment D and F are 235 days and 310 days, and exposure period in $-2.5^{\circ}C$...was longer than other treatments. Numbers of hatched lines of treatment D and F are 48 and 29, and occupied 15.6% and 9.4% of all tested lines, respectively. Average hatching ratio of treatment D and F were 54.% and 71.6%, and average dead egg ratio were 12.6% and 2.4%, respectively. These results show that average ratio of hatching dead eggs in treatment D and F are higher than general line. Thus reconsideration of hatching condition on treatment D and F is needed.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.466-473
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• 2006

Impaired insulin secretion from pancreatic beta-cells in response to glucose is an important feature in the pathology of non-insulin-dependent diabetes mellitus (NIDDM). In the course of screening for useful insulin secretagogues, we have isolated and identified YHB-2017 (Genistein) as a insulin secretion potentiator from fermentation broths of our in-house microbial library. The insulinotropic activity of YHB-2017 in isolated rat pancreatic islets was exerted only at high concentration of glucose (8.3-16 mM) but not at low concentration of glucose (3.3-5.5 mM). Also, in perifusion study with isolated rat pancreatic islets, YHB-2017 stimulated insulin secretion in a time-dependent manner when YHB-2017 was added to KRB buffer containing 16 mM glucose. In the presence of $200\;{\mu}M$ diazoxide and 35 mM KCI, which stimulates maximum $Ca^{2+}$ influx independently of KATP channel, YHB-2017 enhanced KATP channel-independent insulin secretion at high concentration glucose (16 mM). To elucidate the mechanisms of the glucose-dependent potentiation effect of YHB-2017, pharmacologic inhibitors for protein kinase A, protein kinase C and calcium/calmodulin kinase II were pre-treated and then the potentiation effect of YHB-2017 on insulin secretion was investigated. Pre-treatment of H89 as a PKA inhibitor had a significant inhibitory effect on YHB-2017-induced potentiation effect. Furthermore, western immunoblotting analyses revealed that YHB-2017 increased phosphorylation of PKA substrates and cAMP response element-binding protein (CREB) under high concentration of glucose. These results demonstrated that the insulinotropic effect of YHB-2017 is mediated through PKA signal pathway and activated amplifying $K_{ATP}$ channel-independent insulin secretion pathway.

• Journal of the Korean earth science society
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• v.26 no.1
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• pp.93-105
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• 2005

Climatological characteristics in the variation of soil temperatures in Korea were investigated using Korea Meterological Administration's observation data. And the impacts of soil moisture on the variation of soil temperature were examined using observed precipitation data. The climatological averages of soil temperatures are ranged from 14.4 to $15.0^{\circ}C$ regardless of depths. And they showed an latitudinal gradient with a warm temperature at the southern region and 'U' shape as in the air temperature with a high value along the coastal region. The relatively higher heat capacity and low conductivity of soil compared to those of the air resulted in the significant delay of the maximum and minimum date with depth. As a results, soil acts as a heat source during winter while a heat sink during summer. Global warming and urban heat island have increased the soil temperatures with an average rate $0.3\~0.5^{\circ}C/10-year$ as in the air temperature during last 30 years $(1973\~2002)$. However, the warming rate is maximized during spring contrary to the winter in the air temperature. The temporal variation of soil temperatures is strongly affected by that of soil moisture through an modification of the heat capacity and heat convection. In general, the increased soil moisture clearly decreased the temporal variations and increased the deep layer soil temperatures during cold season.