• 한국수산과학회지
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• 제56권4호
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• pp.520-525
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• 2023

This study aimed to detect Enterococcus spp. strain, a fecal contamination indicator, by PCR assay from short neck clams Venerupis philippinarum in Cheonsu Bay area, Chu Island area and Wonsan Island area, the west coast of Korea, from November 2022 to February 2023 of Enterococcus spp. strain was detected in 19 (79.2%) among 24 samples, and its concentration ranged from <18 to 33,000 MPN (most probable number)/100 g. The 269 isolated Enterococcus spp. strains were identified by PCR assay, and Enterococcus spp. distribution in short neck clams were E. faecium (39.8%), E. faecalis (23.0%), E. hirae (21.9%), E. gallinarum (10.4%), E. casseliflavus (1.5%), E. durans (1.5%) and unidentified strains (1.9%). Thus, E. faecium was the most dominant strain followed by E. faecalis. Overall, these results provide novel insight into the necessity for shellfish sanitation in the sea and could help reduce the fecal contamination risk.

• 대한수의학회지
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• 제40권1호
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• pp.81-85
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• 2000

Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

• 치과방사선
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• 제29권1호
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• pp.105-117
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• 1999

Purpose: Ionizing radiations have been reported as an apoptosis initiating stimulus in various cells and it has established that sustained elevations in [Ca/sup 2+/] can lead to DNA fragmentation by Ca/sup 2+/-dependent endonucleases, ultimately resulting in apoptotic cell death. The previous experiments have been reported by using primarily thymocytes and lymphocytes and the change of [Ca/sup 2+/] was measured only by minutes or hours respectively. We need to evaluate [Ca/sup 2+/] in both several minutes and hours after irradiation of radiation of radiation therapy and verify the apoptotic cells. Materials and Methods: We have measured [Ca/sup 2+/] in human gingival epitheloid cancer cell with 10Gy irradiation, at minutely intervals and hourly intervals using digitized video-intensified fluorescence microscopy and the fluorescent Ca/sup 2+/ indicator dye, fura-2. In order to find out that the transient rise in [Ca/sup 2+/] could induced apoptosis, cells were incubated for 1 hour at 37℃ with TdT enzyme, rinsed and resuspended containing fluorescence and observed under a confocal fluorescence microscope. MTT assay was done to determine cell activity and LDH assay was done to determine the amount of necrotic cells. Results: After irradiation, the transient and temporal increasing of [Ca/sup 2+/] in the KB cells was founded. Though, there was no change in the intracellular [Ca/sup 2+/] at 30 minutes and 2 hours after irradiation. We could detect of DNA fragmented cells at 4 hours after 10Gy irradiated cells. There were no significant differences between 4 hour, 1 day, 3 day cells. There were no significant differences in MTT and LDH assay between the irradiated group and the control group after 4 hours and 1 day. Though after 3 days there were differences in MTT and LDH assay between the irradiated group was significantly decreased than the control group, in LDH assay the number of necrotic cell death of the irradiated was higher than the control group. Conclusion: In KB cells there were incipient and temporal increasing of the [Ca/sup 2+/] with 10Gy irradiation and the apoptosis was founded from 4 hours later which was earlier than seeing of the change of the amount of the cellular ability and necrosis.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1706-1712
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• 2006

Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

• Food Science and Biotechnology
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• 제16권1호
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• pp.99-103
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• 2007

The purpose of this study was to develop a simple detection method, possibly at the species-level, that allows for large-scale screening of bifidobacteria. Human fecal samples were plated on MRS-raffinose agar containing cysteine and neomycin sulfate, serving as selective pressure for bifidobacteria, and 0.003%(w/v) bromocresol green. All of the test strains grew well on this medium at $37{\pm}1^{\circ}C$, forming white colonies surrounded by yellow halos, which presented a sharp contrast against the green background. In this disc assay, the required incubation time to develop a yellowish zone varied with the species of Bifidobacterium that was tested, allowing for differential counts and easy identification at the species-level: 10-14 hr for B. bifidum, 20-22 hr for B. catenulatum and B. infantis. and 24-25 hr for B. longum and B. breve. No apparent color was observed for B. angulatum and B. adolescentis 28 hr after inoculation. To evaluate the results of pH indicator-based identification, individual isolates were subjected to a colony-PCR experiment with genus-specific primers. The amplified products from the isolates were in good accordance with those from the reference strains at a level of 95% agreement. These results suggest that the present method could be conveniently applied to cell counts, as well as to the preliminary identification of bifidobacteria from a variety of sample types including human feces, dairy products, and commercial probiotic supplements.

• 한국수산과학회지
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• 제26권3호
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• pp.205-213
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• 1993

APHA-MPA(American Public Health Association-Most Probable Numbers)은 환경수중에 존재하는 위생지표세균 추정에 사용되는 미국 FDA의 유일한 공인방법이기는 하나 결과를 얻기까지 많은 시간, 인력 및 경비가 소요될 뿐 아니라 일반적으로 정밀도도 낮다는 결점이 있다. 이러한 문제를 극복하기 위하여 개발된 막여과 방법중 mTEC(membrane thermotolerant E. coli)은 분변계대장균 및 E. coli의 회수율이 좋을 뿐 아니라 경비도 절약할 수 있는 방법으로서 알려져 있어, 미국 Rhode Island주의 Greenwich Bay의 위생조사를 통하여 MPN과 비교하여 보았다. 그 결과 mTEC의 분변계 대장균 및 E. coli 회수율은 MPN보다 평균치로써 각각 1.08 및 1.27배 높게 나타나, MPN의 대체 방법으로서의 가능성이 확인되었으며, 따라서 이 방법이 앞으로 위생지표세균의 검출방법으로서 공인된다면 우리나라 수출용 패류생산 지정해역의 관리는 물론 패류양식장의 위생학적인 분류도 효율적으로 할 수 있으리라 기대된다.

• 대한임상검사과학회지
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• 제48권3호
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• pp.176-182
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• 2016

대식세포에서 염증반응이 진행되면, interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) 등의 cytokine들이 발현되어 inducible nitric oxide synthase (iNOS), prostaglandin E2(PGE2) 등의 염증유발인자가 생성된다. 오가피, 우슬, 두충 각각의 추출물이 어느 정도의 항 염증 효능을 보이며 어떤 pro-inflammatory cytokine의 발현을 억제하는지에 대한 연구를 진행하였다. 오가피, 우슬, 두충은 물 추출하고 동결 건조시켰다. 각각의 추출물의 구성 성분들이 잘 추출되었는지 확인하기 위하여 지표물질인 acanthoside D, 20-hydroxyecdysone, pinoresinol diglucoside를 HPLC로 분석하였다. 항 염증 효능을 확인하기 위하여 lipopolysaccharide (LPS)로 RAW 264.7 세포주를 자극하여 염증 반응을 일으킨 상태에서 각각의 추출물을 농도 별로 처리하고 NO assay를 통해 항 염증 효능을 확인하였으며 real time PCR로 pro-inflammatory cytokine들의 발현량을 측정하였다. 결과적으로 각각의 추출물은 지표성분들이 검출되었으며 오가피와 우슬이 두충보다 NO assay에서 높은 활성을 보였다. Cytokine 발현량 측정에서는 오가피와 우슬은 iNOS와 IL-6의 발현을 억제하였고, 우슬은 TNF-${\alpha}$의 발현을 억제하였다. 우리나라는 전통적으로 약재를 조합하여 처방하여 왔다. 본 연구는 관절염에 전통적으로 사용해 오던 약재들이 어떤 기전에 의하여 항 염증 반응을 보이는지 확인하고 이들을 조합하여 사용하였을 때 어떤 근거에 의하여 시너지 효능을 보이는지 확인하였다.

• Journal of Yeungnam Medical Science
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• 제29권2호
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• pp.83-88
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• 2012

Background: This study was conducted to evaluate the usefulness of the BACTEC MGIT (Mycobacterium Growth Indicator Tube) 960 system for mycobacteria culture and immunochromatographic assay to identify Mycobacterium tuberculosis (MTB) in positive MGIT culture. Methods: Mycobacteria-culture-positive cases were retrospectively analyzed from December 2010 to July 2011. The detection rates and the recovery times of the mycobacteria between the Ogawa media and the MGIT were compared. An immunochromatographic assay (ICA) (SD BIO-LINE) was also performed in the positive MGIT culture for identification, and the results were compared with those of the Ogawa media in the Korea National Tuberculosis Association. Results: Among the 261 patients (M:F, 168:93; mean age, $61.6{\pm}17.16$ yrs), 450 specimens (sputa, 365; bronchial washing, 61; and pleural effusion, 24) were found positive with mycobacteria. Mycobacteria were grown both on the MGIT and Ogawa media in 310 cases (68.9%); only on the MGIT in 115 cases (22.6%); and only on the Ogawa media in 25 cases (5.5%) (p<0.05). The recovery time was $28.2{\pm}8.9$ days in the Ogawa media and $11.1{\pm}5.8$ days in the MGIT (p<0.05). Among the 127 cases from the positive MGIT culture, all 92 cases that were confirmed as MTB cases bythe Korea National Tuberculosis Association were identified as MTB by ICA, with 100% sensitivity. Conclusion: MGIT increases the detection rate and shortens the recovery time of mycobacteria in clinical respiratory specimens, and the TB Ag MPT64 kit using ICA is useful in identifying MTB in a positive MGIT culture.

• 대한핵의학회지
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• 제19권1호
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• pp.119-126
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• 1985

To evaluate the performance characteristics of CA 19-9 radioimmunoassay and the clinical significance of serum CA 19-9 assay in patients with malignancy, serum CA 19-9 levels were measured by radioimmunoassay using monoclonal antibody in 135 normal controls, 81 patients with various untreated malignancy, 9 patients of postoperative colon cancer without recurrence and 20 patients with benign gastrointestinal diseases, who visited Seoul National University Hospital from June, 1984 to March, 1985. The results were as follows; 1) The CA 19-9 radioimmunoassay was simple to perform and can be completed in one work day. And the between-assay reproducibility and the assay recovery were both excellent. 2) The mean serum CA 19-9 level in 135 normal controls was $8.4{\pm}4.2U/mL$. Normal upper limit of serum CA 19-9 was defined as 21.0 U/mL. 4 out of 135(3.0%) normal controls showed elevated CA 19-9 levels above the normal upper limit. 3) One out of 20(5.0%) patients with benign gastrointestinal diseases showed elevated serum CA 19-9 level above the normal upper limit. 4) In 81 patients with various' untreated malignancy, 41 patients(50.6%) showed elevated serum CA 19-9 levels. 66.7% of 18 patients with colorectal cancer, 100% of 2 patients with pancreatic cancer, 100% of 3 patients with common bile duct cancer, 47.1% of 17 patients with stomach cancer, 28.6% of 28 patients with hepatoma and 60.0% of 5 other gastrointestinal tract cancers showed elevated serum CA 19-9 levels. 5) The sensitivities of serum CA 19-9 related to resectability in colorectal and stomach cancer were 33.3% in resectable colorectal cancer, 83.3% in unresectable colorectal cancer, 41.7% in resectable stomach cancer, 60.0% in unresectable stomach cancer respectively. 6) The sensitivity of serum CA 19-9 in 9 patients of postoperative colorectal cancer without recurrence were 33.3% and significantly decreased compared with that of untreated colorectal cancer, 66.7% (p<0.05). 7) In patients with colorectal cancer, simultaneous measurement of serum CA 19-9 and serum CEA levels increased sensitivities. From above results, we concluded that serum CA 19-9 radioimmunoassay is simple to perform and reproducible, and is a useful indicator reflecting tumor extent and responses to the treatment in patients with malignancy.

• 한국수정란이식학회지
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• 제27권2호
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• pp.93-100
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• 2012

To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.