• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.88-89
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• 2013

A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

• Journal of Veterinary Clinics
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• v.19 no.4
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• pp.418-425
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• 2002

Estrus was induced in 13 anestrus Korea Jin-do bitches by intramuscular injection of pregnant mare serum gonadotropin (PMSG) in a dose of 500 lU once daily for ten consecutive days, followed by an additional single intraveneous injection of 1,000 lU of human chorionic gonadotropin (hCG) on the tenth day. Day-changes of vaginal epithelial cells during the hormone treatment were investigated in each experimental bitches and compared with the those of spontaneous estrus bitches. The first days of vulval bleeding and male acceptance after PMSG treatment were on Day 6.0$\pm$ 1.5 (mean$\pm$ SD) and Day 9.0$\pm$ 1.9, respectively. And in all of 13 bitches, vulval swelling and perineal reflex were shown. The mean durations of proestrus and estrus were 2.9$\pm$ 1.4 (mean$\pm$ SD, range ; 1-6) and 11.5: 1.7 (range ; 8-14) days, respectively, that is, duration of proestrus was significantly shorter than that of the spontneous estrous bitches but duration of estrus was longer than that of the spontaneous estrous bitches. Characteristic features of vaginal cytology during the estrous cycle were the high proportions of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus and parabasal cell, small intermediate, large intermediate cell, and leukocyte in diestrus, respectively. The comification index (Cl) was significantly high proportion in proestrus and estrus, when Day 0 was timed from the first day of male acceptance, the Cl was first increased above 80% on Day 0 and maintained above 80% until Day 0 to Day 5 during 6 days and showed a peak on Day 2. Also it was maintained above 90% until Day 2 to Day 3 during 2 days. These results indicated that all 13 ekperimental bitches showed positive estrus detection by the estrus behavior and vaginal smear test after treated with PMSG and hCC. It suggested that vaginal cytology was used to estimate the optimal mating and ovulation time, in consideration of the day when the Cl was maintained above 80% in estrus-induced Korea Jin-do bitches.

• Bulletin of the Korean Chemical Society
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• v.21 no.5
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• pp.483-486
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• 2000

A method is described for the fluorimetric determination of nickel, based on the formation of $Ni(II)-\alpha-(2-Benzimidazolyl)-\alpha'$, $\alpha''$ -(N-5-Nitro-2-Pyridylhydrazone)-toluene complex in the presence of a non-ionic surfactant. The complex has practically no fluorescence in the absence of surfactant, but the addition of Triton X-100 makes possible the fluorimetric determination of low concentrations of Ni(II) as it enhances the fluorescenceintensity of the complex by up to about 5-fold. This method is very sensitive and selectrive for the direct determination of nickel ion. The optimum conditions are a Triton X-100 concentration of 2.0 mL(5.0%, v/v) and pH $9.0\pm0.2(ammonium$ chloride-ammonia buffer). The fluorescence is measured at 337 nm of emission wavelength under 300 nm of excitation wavelength. The fluorescence intensity is a linear function of the concentration of Ni(II) in the range 5-70 ng/mL, and the detection limit is 2.0 nm/mL. The proposed method has been successfully applied to the determination of trace amounts of Ni(II) in food and human hair samples.

• Journal of Korean Society of Water and Wastewater
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• v.32 no.2
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• pp.131-143
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• 2018

Phosphorus (P) is a limited, essential, and irreplaceable nutrient for the biological activity of all the living organisms. Sewage sludge ash (SSA) is one of the most important secondary P resources due to its high P content. The SSA has been intensively investigated to recover P by wet chemicals (acid or alkali). Even though $H_2SO_4$ was mainly used to extract P because of its low cost and accessibility, the formation of $CaSO_4$ (gypsum) hinders its use. Heavy metals in the SSA also cause a significant problem in P recovery since fertilizer needs to meet government standards for human health. Therefore, P recovery process with selective heavy metal removal needs to be developed. In this paper some of the most advanced P recovery processes have been introduced and discussed their technical characteristics. The results showed that further research is needed to identify the chemical mechanisms of P transformation in the recovery process and to increase P recovery efficiency and the yields.

• The Korean Journal of Pharmacology
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• v.31 no.1 s.57
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• pp.103-114
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• 1995

Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as renal toxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and improving antitumor activity. We synthesized new Pt (Ⅱ) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,2-bis(diphenylphosphino) ethane (DPPE) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of [Pt(trans-ddach)(DPPE).$2NO_3(PC)$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC demonstrated acceptable antitumor activity aganist P388, L-1210 lymphocytic leukemia cells and SK=OV3 human ovarian adenocarcinoma cells, and significant. activity as compared with that. cisplatin. The toxicity of PC was found quite less than thar of cisplatin using MTT, $[^3H]$ thymidine uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.18-18
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• 2023

Two new phenanthrenes, namely, (1R,2R)-1,7-hydroxy-2,8-methoxy-2,3-dihydrophenanthrene-4(1H)-one (1) and 2,7-dihydroxyphenanthrene-1,4-dione (2), were isolated from the ethyl acetate-soluble fraction of Dendrobii Herba, together with the eleven known compounds, densiflorol B (3), 6,7-dimethoxyphenanthrene2,5-diol (4), dehydroorchinol (5), 1,5,7-trimethoxy-2-phenanthrenol (6), denthyrsinin (7), ephemerantol A (8), lusianthridin (9), moscatilin (10), gigantol (11), 3-[(1E)-2-(3-hydroxyphenyl)ethenyl]-5-methoxyphenol (12) and (-)-syringaresinol (13). Structures of 1 and 2 were elucidated by analyzing 1D and 2D NMR as well as HR-ESI-MS data. The absolute configuration of compound 1 was confirmed by ECD spectroscopic method. In cytotoxicity evaluation using FaDu human hypopharyngeal squamous carcinoma cell line, compounds 3-6, 8, 10 and 12 showed activities, with IC₅₀ values ranging from 2.55 to 17.70 μM. Among them, compound 10 exhibited remarkable cytotoxic activity with IC₅₀ value of 2.55 μM. This is the first report on the anticancer mechanistic study of compound 10 in HNSCC including FaDu cells.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.1
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• pp.101-108
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• 1998

We investigated the effect of ${\alpha}-subunit$ of the stimulatory GTP-binding protein ($G{\alpha}_s$) gene mutation on the expression of the thyrotropin-releasing hormone (TRH) receptor (TRH-R) gene in GH3 cells and in growth hormone (GH)-secreting adenomas of acromegalic patients. In the presence of cyclohexicmide, forskolin and isobutylmethylxanthine, cholera toxin, and GH-releasing hormone (GHRH) decreased rat TRH-R (rTRH-R) gene expression by about 39%, 43.7%, and 46.7%, respectively. Transient expression of a vector expressing mutant-type $G{\alpha}_s$ decreased the rTRH-R gene expression by about 50% at 24 h of transfection, whereas a wild-type $G{\alpha}_s$ expression vector did not. The transcript of human TRH-R (hTRH-R) gene was detected in 6 of 8 (75%) tumors. Three of them (50%) showed the paradoxical GH response to TRH and the other three patients did not show the response. The relative expression of hTRH-R mRNA in the tumors from patients with the paradoxical response of GH to TRH did not differ from that in the tumors from patients without the paradoxical response. Direct PCR sequencing of $G{\alpha}_s$ gene disclosed a mutant allele and a normal allele only at codon 201 in 4 of 8 tumors. The paradoxical response to TRH was observed in 2 of 4 patients without the mutation, and 2 of 4 patients with the mutation. The hTRH-R gene expression of pituitaty adenomsa did not differ between the tumors without the mutation and those with mutation. The present study suggests that the expression of TRH-R gene is not likely to be a main determinant for the paradoxical response of GH to TRH, and that $G{\alpha}_s$ mutation may suppress the gene expression of TRH-R in GH-secreting adenoma. However, a certain predisposing factor(s) may play an important role in determining the expression of TRH-R.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.782-787
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• 2015

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah^-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

• Fisheries and Aquatic Sciences
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• v.26 no.1
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• pp.48-57
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• 2023

One of the ways to increase the production for aquaculture is through the cultivation of monosexuals by ensuring genital reversal from which energy for reproduction is diverted towards growth. Masculinization has been identified as one of the most prominent techniques, where sex development was directed from female to male. This approach only altered the phenotype and not the genotype. The red claw crayfish (Cherax quadricarinatus) was a relatively new commercial commodity, and the males were known to grow faster than females. Hence, it was proposed to use monocultures comprising an all-male population to increase yield using steroid hormone, synthetic 17α-methyltestosterone. However, this technique generated residues that detrimentally affect human health, the environment, and cultivated organisms. Therefore, finding new safe natural steroid sources was essential, and one of which is exploring of natural hormones extracted from the viscera of sea cucumbers (Holothuria scabra Jaeger). This study focused on the determination of male formation and testosterone levels among juvenile crayfish, after immersing in sea cucumber steroid extract (SCSE). A completely random design with factorial was used with two variables, encompassing the varied doses (0, 2, 4 mg/L, 2 mg/L 17α-methyl testosterone as control group) and immersion times of 18 and 30 h. The result showed the dose-dependent ability of SCSE increase the male genital formation and promote the testosterone level of juvenile crayfish. In addition, the testosterone was influenced by dose and immersion duration time, with the highest level of testosterone observed in treatments of 4 mg/L SCSE with 30 h immersion was 0.248 ng/mL, while the male percentage was 77%. In conclusion, the combination of dose and immersion time significantly affected growth and testosterone levels.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7053-7060
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• 2015

In the present work, novel orange peel was extracted with 100%EtOH (ethanol) and fractionated into four fractions namely F1, F2, F3, F4 which were eluted from paper chromatographs using 100%EtOH, 80%EtOH, 50%EtOH and pure water respectively. The crude extract and its four fractions were evaluated for their total polyphenol content (TPC), total flavonoid content (TFC) and radical scavenging activity using DPPH (1,1-diphenyl-2-picrylhydrazyl) assay. Their cytotoxic activity using WST assay and DNA damage by agarose gel electrophoresis were also evaluated in a human leukemia HL-60 cell line. The findings revealed that F4 had the highest TPC followed by crude extract, F2, F3 and F1. However, the crude extract had the highest TFC followed by F4, F3, F2, and F1. Depending on the values of $EC_{50}$ and trolox equivalent antioxidant capacity, F4 possessed the strongest antioxidant activity while F1 and F2 displayed weak antioxidant activity. Further, incubation HL-60 cells with extract/fractions for 24h caused an inhibition of cell viability in a concentration-dependent manner. F3 and F4 exhibited a high antiproliferative activity with a narrow range of $IC_{50}$ values ($45.9-48.9{\mu}g/ml$). Crude extract exhibited the weakest antiproliferative activity with an $IC_{50}$ value of $314.89{\mu}g/ml$. Analysis of DNA fragmentation displayed DNA degradation in the form of a smear-type pattern upon agarose gel after incubation of HL-60 cells with F3 and F4 for 6 h. Overall, F3 and F4 appear to be good sources of phytochemicals with antioxidant and potential anticancer activities.