• Title/Summary/Keyword: Detection,

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Chromosomal Localization and Mutation Detection of the Porcine APM1 Gene Encoding Adiponectin (Adiponectin을 암호화하는 돼지 APM1 유전자의 염색체상 위치파악과 돌연변이 탐색)

  • Park, E.W.;Kim, J.H.;Seo, B.Y.;Jung, K.C.;Yu, S.L.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.537-546
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    • 2004
  • Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.

Perspective on Rapid and Selective Method for Detecting Microbiology in Dairy Industry: A Review (낙농산업에 필요한 미생물 검사방법과 전망: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Kim, Dong-Hyeon;Song, Kwang-Young;Yim, Jin-Hyuk;Choi, Dasom;Lim, Jong-Soo;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • 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.

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Temporal Patterns of Pesticide Residues in the Keum, Mangyung and Dongjin Rivers in 2002 (2002년 금강, 만경-동진강 하천수 중 잔류농약의 연간 검출 양상)

  • Kim, Chan-sub;Lee, Hee-Dong;Ihm, Yang-Bin;Son, Kyeong-Ae
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.230-240
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    • 2017
  • BACKGROUND: To evaluate residues of environmentally concerned pesticides in water system, this monitoring was conducted over three rivers. The residual characteristics and discharging condition of these residues on water system was investigated. METHODS AND RESULTS: Total twenty nine sampling sites were selected through main streams and branch streams of Keum, Mangyung and Dongjin rivers, and the water samples from them were regularly collected one month interval, especially biweekly from May to August in 2002. Of the pesticides monitored, six fungicides which include hexaconazole, isoprothiolane and iprobenfos were detected with frequencies of 0.3-50.9% and in their residue level of $0.1-4.7{\mu}g/L$. Sixteen insecticides which include nine organophosphoruses, three carbamates, endosulfan, cypermethrin, buprofezin and fipronil were detected with frequencies of 0.3-32.5% and in their residue level of $0.01-2.8{\mu}g/L$. Nine herbicides which include alachlor molinate, anilofos, butachlor, dimepiperate, metolachlor, oxadiazon, pretilachlor and thiobencarb were detected with frequencies of 0.8-22.9% and in their residue level of $0.01-9.07{\mu}g/L$. CONCLUSION: Detection frequencies and residue levels of insecticides and herbicides were the highest in waters sampled in May and June. Almost pesticides detected were for the paddy rice and their residue levels were very low to compare with standard values.

Comparison of Flavonoid Characteristics between Blueberry (Vaccinium corymbosum) and Black Raspberry (Rubus coreanus) Cultivated in Korea using UPLC-DAD-QTOF/MS (UPLC-DAD-QTOF/MS를 이용한 국내 재배 블루베리(Vaccinium corymbosum)와 복분자(Rubus coreanus)의 플라보노이드 특성 비교)

  • Jin, Young;Kim, Heon-Woong;Lee, Min-Ki;Lee, Seon-Hye;Jang, Hwan-Hee;Hwang, Yu-Jin;Choe, Jeong-Sook;Lee, Sung-Hyun;Cha, Youn-Soo;Kim, Jung-Bon
    • Korean Journal of Environmental Agriculture
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    • v.36 no.2
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    • pp.87-96
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    • 2017
  • BACKGROUND: The objective of this study was to identify and compare the main phenolic compounds (anthocyanins, flavonoids, phenolic acids) in blueberry and black raspberry cultivated in Korea using ultra-performance liquid chromatography diode array detection-quadrupole time-of-flight mass spectrometry (UPLC-DAD-QTOF/MS). METHODS AND RESULTS: Twenty-nine flavonoids were identified by comparison of ultraviolet and mass spectra with data in a chemical library and published data. Blueberry contained flavonols including kaempferol, quercetin, isorhamnetin, myricetin, and syringetin aglycones. Isorhamnetin 3-O-robinobioside, kaempferol 3-O-(6"-O-acetyl)glucoside, quercetin, quercetin 3-O-arabinofuranoside (avicularin), quercetin 3-O-(6''-O-malonyl) glucoside, and quercetin 3-O-robinobioside were detected for the first time in blueberry. The flavonoids in raspberry consisted of quercetin aglycone and its glycosides. The mean total flavonoid content in blueberry [143.0 mg/100 g dry weight (DW)] was 1.5-times that in raspberry (95.4 mg/100 g DW). The most abundant flavonoid in blueberry was quercetin 3-O-galactoside (hyperoside, up to 76.1 mg/100 g DW) and that in raspberry was quercetin 3-O-glucuronide (miquelianin, up to 55.5 mg/100 g DW). Miquelianin was not detected in blueberry. CONCLUSION: Flavonol glycosides were the main flavonoids in blueberry and black raspberry cultivated in Korea. The composition and contents of flavonoids differed between blueberry and black raspberry, and may be affected by the cultivar and cultivation conditions.

Arsenic Speciation and Risk Assessment of Miscellaneous Cereals by HPLC-ICP-MS (HPLC-ICP-MS를 활용한 잡곡의 비소 화학종 및 위해 분석)

  • An, Jae-Min;Hong, Kyong-Suk;Kim, Sung-Youn;Kim, Dae-Jung;Lee, Ho-Jin;Shin, Hee-Chang
    • Korean Journal of Environmental Agriculture
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    • v.36 no.2
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    • pp.119-128
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    • 2017
  • BACKGROUND: Miscellaneous cereal have been largely consumed in Korea as due to their physiological functions beneficial to human health. The cereals are currently a social concern because they have been found to contain heavy metals. Thus, monitoring heavy metals in the cereals is an important requirement for food safety analysis. In this study, we determined arsenic concentration in the cereals randomly harvested from different markets. METHODS AND RESULTS: Inorganic arsenic was determined by ICP-MS coupled with HPLC system. The HPLC-ICP-MS analysis was optimized based on the limit of detection and recover test to reach $0.13-1.24{\mu}g/kg$ and 94.3-102.1%, respectively. The concentrations of inorganic arsenic equivalent to daily exposure were levels of $19.91{\mu}g/day$ in mixed grain, $1.07{\mu}g/day$ in glutinous rice, $0.77{\mu}g/day$ in black brown rice, $0.13{\mu}g/day$ in barley and $0.11{\mu}g/day$ in soybeans. CONCLUSION: The levels of arsenic in miscellaneous cereals were found lower than the recommended The Joint FAO/WHO Expert Committee on Food Additives (JECFA) levels, suggesting that the cereals marketed in Korea are not potential concern in risk assessment.

Dynamin II Expression and Morphological Comparison of NIH3T3 and NIH3T3 (ras) Cells (NIH3T3와 NIH3T3(ras) 세포에서 Dynamin II 발현 및 형태적 비교)

  • Lee, Chul-Woo;Kim, Su-Gwan;Choi, Jeong-Yun;Choi, Baik-Dong;Bae, Chun-Sik;Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.35 no.3
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    • pp.121-128
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    • 2005
  • It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.

Histochemical Detection of Ionic Zinc in the Rat Olfactory Mucosa: Zinc Selenium Autometallography ($ZnSe^{AMG}$) (랫드 후각점막내 Zinc 이온의 조직화학적 동정)

  • Nam, Dong-Woo;Sun, Yuan-Jie;Kim, Sung-Joo;Kim, Yong-Kuk;Kim, Soo-Jin;Yu, Yun-Cho;Jeong, Young-Gil;Jo, Seung-Mook
    • Applied Microscopy
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    • v.33 no.2
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    • pp.145-154
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    • 2003
  • The present study was designed to demonstrate ionic zinc in the rat nasal mucosa by means of zinc selenium autometallography ($ZnSe^{AMG}$). Rats were given sodium selenide either intraperitoneally (i.p) or intranasally (i.n). Prior to the i.n. administration the rats were anesthetized with pentobarbital sodium (30 mg/kg, i.p.). A thin plastic tube coupled to a Hamilton syringe was then inserted into the right nostril and $10{\mu}l$ of the solution was instilled. For the i.p. administration non-anesthetized rats were given $100{\mu}l$ of the sodium selenide solution (10 mg/kg). Control rats were instilled with saline. After 2 hrs survival, the rats were anaesthetized and transcardially perfused with 3% glutaraldehyde. The olfactory area was removed and put into same fixative. The nose was then sectioned ($30{\mu}m$) horizontally, autometallography (AMG) was performed according to Danscher et al. (1997). After silver enhancement, fine AMG grains were scattered in the whole length of the olfactory epithelium containing olfactory receptor neurons, sustentacular and basal cells. However, much higher concentration of the AMG grains occupied near the surface and in the basal region of the olfactory epithelium. Both groups of i.p. and i.n. administration showed almost same level in the concentration of the AMG grains. In i.n. group, few AMG grains were also found in olfactory nerves of the lamina propria, suggesting zinc transport into the olfactory bulb via olfactory axons. At the electron microscopic level, the AMG grains were most entirely found in the supporting cells of the olfactory epithelium, and they were mostly localized in lysosome-like organelles. The i.n. group showed various signs of tissue damage of the olfactory mucosa, where dense concentration of AMG grains were localized at crystalloid structures. The present study demonstrated dense population of ionic zinc in the rat olfactory epithelium. zinc may play a role in the olfactory functioin and in the pathogenesis of the neurodegerative disorders affecting nose.

Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

Evaluation of p16INK4a/Ki-67 Dual Immunostaining in Liquid-based Cytology for Diagnosis of Uterine Cervical Dysplasia and Cancer (자궁경부 이형성증과 암의 진단을 위한 액상세포 검체에서 p16INK4a/Ki-67 이중면역염색의 평가)

  • Sung, Mi Hee;Lee, Hoon Taek;Shin, Min Shik;Oh, Seo Young;Kim, Wook Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.132-139
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    • 2015
  • Recently, $p16^{INK4a}$/Ki-67 dual immunostaining has been introduced as a new biomarker protocol for early detection of uterine cervical dysplasia and cancer in liquid-based cytology (LBC). We performed the $p16^{INK4a}$/Ki-67 dual immunostaining using a CINtec$^{(R)}$ PLUS kit in a total of 109 LBC cases of cervicovaginal smear and compared its results with those from LBC, HPV hybrid capture II (HC II) test and histological diagnosis. Expression of $p16^{INK4a}$ and Ki-67 was significantly associated with cases of LSIL or higher in cytological diagnosis and cases of cervical intraepithelial neoplasia (CIN) 1 or higher in histological diagnosis (p<0.001 and p<0.001, respectively). Among forty-six cases of atypical squamous cells of undetermined significance (ASCUS) in LBC, $p16^{INK4a}$ and Ki-67 was expressed in 31 (67.4%), which were positively associated with cases of CIN I lesion or higher in histology. The sensitivity of $p16^{INK4a}$/Ki-67 dual immunostaining for finding lesions of CIN 1 or higher was 89.0%, which was higher than LBC. The specificity was 73.5%, which was higher than that of the HC II test. Based on these results, the $p16^{INK4a}$/Ki-67 dual immunostaining method can be a useful diagnostic marker for improving the sensitivity of LBC and the specificity of HC II test.

Comparison of an Automated Most-Probable-Number Technique TEMPO®TVC with Traditional Plating Methods PetrifilmTM for Estimating Populations of Total Aerobic Bacteria with Livestock Products (축산물가공품에서 건조필름법과 TEMPO®TVC 검사법의 총세균수 비교분석)

  • Kim, Young-Jo;Wee, Sung-Hwan;Yoon, Ha-Chung;Heo, Eun-Jeong;Park, Hyun-Jeong;Kim, Ji-Ho;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.103-107
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    • 2012
  • We compared between an automated most-probable-number technique $TEMPO^{(R)}$TVC and traditional plating methods $Petrifilm^{TM}$ for estimating populations of total aerobic bacteria in various livestock products. 257 samples randomly selected in local retail stores and 87 samples inoculated with $E.$ $coli$ ATCC 25922, $Staphylococcus$ $aureus$ ATCC 12868 were tested in this study. The degree of agreement was estimated according to the CCFRA (Campden and Chorleywood Food Research Association Group) Guideline 29 and the agreement indicates the difference of two kinds methods is lower than 1 log base 10($log_{10}$). The samples of hams, jerky products, ground meat products, milks, ice creams, infant formulas, and egg heat formed products were showed above 95% in the agreement of methods. In contrast, proportion of agreement on meat extract products, cheeses and sausages were 93.1%, 92.1%, 89.1%, respectively. One press ham and five sausages containing spice and seasoning, two pork cutlets containing spice and bread crumbs, two meat extract product and two natural cheeses and one processing cheese with a high fat content, and one ice cream containing chocolate of all samples showed the discrepancy. Our result suggest that $TEMPO^{(R)}$TVC system is efficient to analyses total aerobic bacteria to compare manual method in time-consuming and laborious process except livestock products having limit of detection.