• Journal of The Korean Society of Grassland and Forage Science
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• v.39 no.4
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• pp.227-234
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• 2019

This study was performed to investigate the effect of heat-stressed environment on rumen microbial diversity in Holstein cows. Rectal temperature and respiration rate were measured and rumen fluid was collected under normal environment (NE; Temperature humidity index (THI)=64.6) and heat-stressed environment (HE; THI=87.2) from 10 Holstein cows (60±17.7 months, 717±64.4 kg) fed on the basis of dairy feeding management in National Institute of Animal Science. The rumen bacteria diversity was analyzed by using the Illumina HiSeq^TM 4000 platform. The rectal temperature and respiratory rate were increased by 1.5℃ and 53 breaths/min in HE compared to that in NE, respectively. In this study, HE exposure induced significant changes of ruminal microbe. At phylum level, Fibrobacteres were increased in HE. At genus level, Ruminococcaceae bacterium P7 and YAD3003, Butyrivibrio sp. AE2032, Erysipelotrichaceae bacterium NK3D112, Bifidobacterium pseudolongum, Lachnospiraceae bacterium FE2018, XBB2008, and AC2029, Eubacterium celulosolvens, Clostridium hathewayi, and Butyrivibrio hungatei were decreased in HE, while Choristoneura murinana nucleopolyhedrovirus, Calothrix parasitica, Nostoc sp. KVJ20, Anabaena sp. ATCC 33047, Fibrobacter sp. UWB13 and sp. UWB5, Lachnospiraceae bacterium G41, and Xanthomonas arboricola were increased in HE. In conclusion, HE might have an effect to change the rumen microbial community in Holstein cows.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.446-454
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• 2017

Human papillomaviruses (HPVs) are major causes of cervical cancer. Sixteen high risk HPVs, including HPV 16, HPV 18, HPV31, HPV 33, HPV 35, HPV 39, HPV 45, HPV 51, HPV 52, HPV 53, HPV 56, HPV 58, HPV 59, HPV 66, HPV 68, and HPV 69 are found in cervical cancer. HPVs 16 and 18 are mainly presented in 70% of cervical cancer. Therefore, identifying the presence of these high-risk HPVs is crucial. The objective of this study is to establish the HPV ViroCheck for detecting 16 HR-HPVs and genotypes of HPVs 16 and 18, as well as to analyze the analytical performance of HPV ViroCheck. We performed the analytical sensitivity of HPV E6 / E7 genes of 16 high risk HPVs to confirm the limit of detection. Then, a cross reactivity of HPV ViroCheck with microorganisms and viruses related to the cervix were analyzed for analytical specificity. Analytical sensitivity of high risk HPV genotypes ranged from 1 to 100 copies when using cloned DNAs. The limit of detection was 10 cells for both SiHa and HeLa cells. Cervical-related microorganisms and viruses did not show cross-reactivity to HPV DNA. Moreover, the intra- and inter-assay coefficient variations (CVs) were below 5%. In conclusion, HPV Virocheck will be useful for the detection of 16 HR HPVs, as well as HPV 16 and HPV 18 genotypes rapidly.

• Journal of Food Hygiene and Safety
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• v.25 no.3
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• pp.269-277
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• 2010

Selenium is an essential micronutrient for normal body function and functions as an essential constituent of selenoproteins. This study was carried out to investigate effect of selenium on the formation of colonic aberrant crypt foci (ACF) and tumor formation in a mouse model. Five-week old ICR mice were acclimated for one week and fed different selenium diet (0.02, 0.1, and 0.5 ppm) for 12 weeks. Animals received three intraperitoneal injections of azoxymethane (10 mg/kg B.W. in saline for 3 weeks), followed by 2% dextran sodium sulfate in the drinking water for a week. There were four experimental groups, including a normal control group and three different selenium levels groups. After sacrifice, the total numbers of aberrant crypt (AC) and ACF were measured in the colonic mucosa after methylene blue staining. The number of tumors was noted for tumor incidence. Liver selenium concentration was measured using ICP-AES method. Gutathione peroxidase (GPx) activity was determined using a GPx assay kit in the liver and colon. TUNEL assay and proliferating cell nuclear antigen (PCNA) staining were performed to examine the cell apoptosis and cell proliferation, respectively. Immunohistochemistry of $\beta$-catenin was also performed on the mucous membrane tissue of colon. The activity of GPx in the liver and colon was decreased in the selenium-deficient diet group while it was increased in the selenium-overloaded diet group. Apoptotic positive cells were increased in the selenium-overloaded diet group but decreased in the selenium-deficient diet group. PCNA staining area was decreased in the selenium-overloaded diet group. In addition, the $\beta$-catenin protein level in the selenium-deficient diet group was increased but decreased in the selenium-overloaded diet group. These results indicate that dietary selenium might exert a modulating effect on colon cancer by inhibiting the development of ACF and colon tumor formation in this mouse model.

• Food Science of Animal Resources
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• v.33 no.3
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• pp.403-410
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• 2013

This study was performed to evaluate the microbiological contamination level of raw beef from retail markets in Seoul, Korea. The sampling and laboratory test were performed according to the procedure of "Standard for processing and ingredients specification of livestock product" and "Korean food code". Enterotoxin of Staphylococcus aureus isolates were detected using VIDAS$^{(R)}$ and PCR-based methods. Listeria monocytogenes serotyping and genotyping were carried out using Listeria antisera and L. monocytogenes Fingerprinting kit, respectively. A total of 48 samples were collected from 16 retail markets (butcher's shop: 5, department store: 6, supermarket: 5) in 2011. The level of total bacteria counts in the butcher's shop, department store and supermarket were $4.4{\times}10^3$ CFU/g, $3.9{\times}10^5$ CFU/g and $1.0{\times}10^4$ CFU/g, respectively. The concentrations of Escherichia coli of these three retail markets were $6.4{\times}10$ CFU/g, 7.6 CFU/g and $2.0{\times}10$ CFU/g, respectively. Salmonella species was not detected on all samples. However, S. aureus was isolated in the 3 samples (6.25%) from each type of three retail markets. L. monocytogenes was isolated in the 4 samples (8.3%) from department stores. The level of contamination of these foodborne bacteria was less than 100 CFU/g. The enterotoxin-encoding genes of S. aureus isolates were sea, seh, sei and sep gene. The gene similarity of L. monocytogenes isolated from two retail markets by Rep-PCR showed 57.8-98.1% and 68.1-98.1%, respectively. These results suggest that the HACCP guideline for environmental control in slaughterhouse and retail markets should be provided to prevent cross contamination and manage foodborne pathogens such as L. monocytogenes and S. aureus.

• The Korean Journal of Nuclear Medicine
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• v.30 no.4
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• pp.541-547
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• 1996

The previous monoclonal antibody labeling method for bone marrow immunoscintigrapy was complicated and laborious for clinical application. Also it showed a relatively low labeling efficiency. To improve this procedure, we compared several direct labeling methods of $^{99m}Tc$. 1) The labeling efficiency in the method using gluconate as a transchelator was low (40-70%), but immunoscintigraphy using this radiotracer produced a clear image. 2) To improve labeling efficiency, ${\beta}$-mercaptoethanol was removed after reduction. The labeling efficiency was improved up to 70-80%, but the radioactivity of the blood pool was high. 3) The higest labeling efficiency (>90%) and best quality images could be obtained by using MDP as a transchelating agent. It did not require additional procedures for separation of labeled antibodies. The immunoreactivity of this antibody was 60%. Residual MDP which can be taken up by the bone could be removed by PD-10 column. The reduced antibodies were stable with a high labeling efficiency (>90%) for up to 47 days by deep freezing. We concluded that the improved procedure for $^{99m}Tc$ labeling of anti-NCA-95 monoclonal antibody using MDP as a transchelating agent will be a simple and useful method for clinical application.

• Journal of Applied Biological Chemistry
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• v.63 no.1
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• pp.95-101
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• 2020

Psoriasis is an autoimmune skin disease that is accompanied by hyper proliferation of the epidermis, erythema of various sizes, and ulceration. However, the mechanism of the development of psoriasis dermatitis is unclear. Recently, it is known that the inflammatory cytokines and Th17 cells as well as chemokine (CC motif) ligand 20 (CCL20) are involved in the process of keratinocytes hyper-differentiation, which is common in psoriasis dermatitis. Therefore, we studied the effects of yakuchinone-A, an active ingredient of Alpinia oxyphylla Miquel known for its anti-inflammatory activity, to improve psoriasis dermatitis. First, cytotoxicity of yakuchinone-A was observed in cell counting kit-8 assay and not observed in 10 ㎍/mL concentration on the human keratinocyte HaCaT cells. Yakuchinone-A in the presence of tumor necrosis factor-alpha (TNF-α) on HaCaT cells inhibited mRNA expression of IL-6, IL-8, and TNF-α by up to 61.4±7.5, 23.6±1.5, 46.0±4.8%. CCL20, a chemokine that attracts immune cells such Th17 cells to the inflammation location, was also significantly suppressed by yakuchinone-A. In addition, IκB and STAT3 phosphorylation involved in the CCL20 expression was inhibited by yakuchinone-A in a concentration-dependent manner up to the level of 79.1±5.0, 80.8±2.3%. Furthermore, yakuchinone-A downregulated CCL20 mRNA expression level on IL-17A-activated HaCaT cells as a concentration-dependent manner. Based on these results, yakuchinone-A is expected to be developed as a new material for improving psoriasis dermatitis in the future.

• Analytical Science and Technology
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• v.17 no.3
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• pp.199-210
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• 2004

A new polystyrene-divinylbenzene chelating resin containing 4,5-dihydroxy-naphthalene-2,7-disulfonic acid (chromotropic acid : CTA) as functional group has been synthesized and characterized. The sorption and desorption properties of this chelating resin for Cr(III) ion and Cr(VI) ion including nine metal bloodstain. As a results, FOB test kit could be effectively applied to identification of human blood at chelating resin was stable in acidic and alkaline solution. The Cr(VI) ion is selectively separated from Cr (III) ion at pH 2 and the maximum sorption capacity of Cr(VI) ion is 1.2 mmol/g. In the presence of anions such as $F^-$, $SO{_4}^{2-}$, $CN^-$, $CH_3COO^-$, $NO{_3}^-$, the sorption of Cr(VI) ion was reduced but anions such as $PO{_4}^{3-}$ and $Cl^-$ revealed no interference effect. The elution order of metal ions obtained from breakthrough capacity and overall capacity at pH 2 was Cr(VI)>Sn(II)>Fe(III)>Cu(II)>Cd(II)${\simeq}Pb(II){\simeq}Cr(III){\simeq}Mn(II){\simeq}Ni(II){\simeq}Al(III)$. Desorption characteristics for Cr(VI) ion was investigated with desorption agents such as $HNO_3$, HCl, and $H_2SO_4$. It was found that the ion showed high desorption efficiency with 3 M HCl. As the result, the chelating resin, XAD-16-CTA was successfully applied to separation and preconcentration of Cr (VI) ion from several metal ions in metal finishing works.

• Journal of Animal Science and Technology
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• v.47 no.5
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• pp.867-876
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• 2005

The microbial contamination of ice cream product commercially available in Korea was determined using ice bar, ice cream, ice milk and non-milk fat ice cream. Irradiation effect on enhancement of microbiological safety was also investigated at doses of 1, 3, and 5 kGy. In all products, yeast and molds were not detected, however, total aerobic and coliform bacteria were detected at 1-2 and 1-1.5 Log CFU/g level, respectively. According to the different flavor used in ice cream, total aerobic bacteria were detected as 2.30, 2.90, and 3.32 Log CFU/g level in vanilla, chocolate, and strawberry ice cream, respectively. Yeast and mold was not detected in vanilla ice cream but 2.30 and 2.70 Log CFU/g in chocolate and strawberry ice cream, respectively. Coliforms were also detected 1-2 Log CFU/g in the ice cream with different flavors. Listeria inocua and Escherichia coli were detected from 3 commercial samples but Salmonella spp. was not detected using API kit. Gamma irradiation significantly reduced the level of the contaminated total aerobic bacteria, yeast and molds and coliform population in the ice creams. These results indicated that irradiation(5kGy or less) is effective to ensure safety of ice cream.

• Journal of Dairy Science and Biotechnology
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• v.33 no.2
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• pp.119-127
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• 2015

To date, detection of microbial populations in dairy products has been performed using culture media, which is a time-consuming and laborious method. The recently developed chromogenic media could be more rapid and specific than classical culture media. However, the newly developed molecular-based technology can detect microbial populations with greater rapidity and sensitivity than the classical method involving culture media and chromogenic media. This molecular-based technology could provide various options for monitoring the characterization of different states of bacteria and cells. Thus, it could help upgrade the processing system of the dairy industry so as to maintain the safety and quality of dairy foods. Among the various newly developed molecular-based technologies, flow cytometry can potentially be used for monitoring microbiological populations in the dairy industry if official international standards are available for this purpose. When omics technology would have biomarker identification, it could be regarded as the rapid and sensitive analytical methods. Methods based on PCR, which has become a basic technique in microbiological research, can be developed and validated as alternative methods for quantification of dairy microorganisms. This review discusses methods for monitoring microbiological populations in dairy foods and the limitations of these studies, as well as the need for further research on such methods in the dairy industry.