• 한국동물위생학회지
• /
• 제17권1호
• /
• pp.37-43
• /
• 1994

The rate of 58 neonatal calves in infection of Theileria sergenti was investigated in random samples on the farms located in Kyunggi, Chonbuk districts of Korea. 1. The criteria used in veryfying infection with T. sergenti included the detection of parasites by giemsa stain and acridine orange stain in the blood smear slides. 2. Further evidence of current or previous exposure to T. sergenti was based on demonstration of T. sergenti specific antibody and antigen by the western immunoblot and the directed immunofluorescent antibody test in the peripherial blood of the calves. 3. The prevalence rates were 35%, 50% in Kyunggi, Chonbuk provinces respectively and the overall prevalence in all the farms was 43.2％ by means of acridine orange stain. 4. The parasites that were observed in the peripherial blood of calves was showen surely by the western immunoblot to the characteristic 34KD antigen among the proteins of T. sergenti (Korean isolate). 5. And the antigen of the neonatal calves reacted at the very highest titer(1 : 2, 560) 6. These data highlight the significances of T. sergenti in the neonatal calf disease in Korea.

• Parasites, Hosts and Diseases
• /
• 제33권4호
• /
• pp.391-394
• /
• 1995

말라리아의 진단 방법으로 흔히 사용되고 있는 Giemsa 염색법과 형광염색법중 Acridine orange(AO) 염색법을 비교하였다. 말라리아 환자의 혈액을 채취하여 Giemsa와 AO로 염색하여 각각 광학현미경 및 형광현미경으로 관찰하였다. AO 염색법은 Giemsa 염색법에 비해 저배율에서도 쉽게 말라리아 원충을 찾을 수 있어 빠르고 정확한 말라리아의 진단을 위해 Acridineorange 염색법이 더 효과적이라고 생각된다.

• 대한수의학회지
• /
• 제33권2호
• /
• pp.289-293
• /
• 1993

Following the death of more than 250 goats in one herd of 800 goats, imported from Australia, an epidemiological investigation was undertaken to determine the probable aetiology of this apprently mysterious disease. The syndrome was characterized by severe anemia(Hematocrit <20% ; normal range 24 to 48). All the affected animals were imported from Australia and all the motalities occurred during the period from September to November, 1992 Giemsa stain, acridine orange and indirect immunoflourescence tests were utilized in a survey involving 239 goats reared in Chonbuk Province. The positivity rates by acridine orange for anaplasmosis or piroplasmosis were 60.8% and 66.2% for imported and indigenous breeds respectively. It is tenatively concluded that the probable cause of death was anaplasmosis.

• 한국임상수의학회지
• /
• 제15권1호
• /
• pp.146-150
• /
• 1998

소 림프구내의 Theileria sergenti감염을 확인하기 위하여 T. sergenti감염혈액에서 림프구를 분리한 후 중합효소연쇄반웅을 실시하였다. 또한,분리한 림프구내의 T. sergenti감 염을 증명하기 위하여 IFA test와 acridine orange stain을 실시하였다. 그 결과 다음과 같은 성적을 얻을 수 있었다. T. sergenti 감염헐액의 전혈과 림프구를 각각 생리식염수로 2배율 연 속회석하여 중합효소연쇄반응을 실시한 결과, 림프구내에서는 1,024배 회석배율까지 T. sergenti의 genomic DNA가 중폭되었으며, 전혈내에서는 256배 회석배율까지 증폭되었다. 그리 고 중합효소연쇄반응으로 T. sergenti 감염이 확인된 림프구를 이용하여 IFA test와 acridine orange 염색을 실시한 결과, 림프구내에 T. sergenti가 존재하는 것을 증명할 수있었다. 한편, 중합효소연쇄반응을 이용한 림프구내의 T. sergenti 감염의 진단 유용성을 확인하기 위하여 전 북지역에서 사육중인 소 16두를 대상으로 이들의 혈액으로 PCR 증폭을 실시하였다. 그 결과 전혈에서 genomic DNA를 취한 경우에는 3두(18.8%)만이, 그리고 림프구에서 genomic DNA를 취한 경우에는 11두(68.8%)의 소에서 T. sergenti DNA의 증폭을 관찰할 수 있었다.

• 대한수의학회지
• /
• 제33권4호
• /
• pp.665-671
• /
• 1993

The rate of 67 neonatal calves's infection of Theileria sergenti was investigated in random samples on the farms located in Kyeongki, Chonbuk and Jeju districts of Korea. The criteria used in verifying infection with T sergenti included the detection of parasites by giemsa's stain and acridine orange stain in the blood smear slides. Further evidence of current or previous exposure to T sergenti was based on the demonstration of T sergenti-specific antibody and antigen by the western immunoblot and the indirect immunofluorescence antibody test in the peripheral blood of the calves. The prevalence rates were 35%, 50% and 100% in Kyeongki, Chonbuk and Jeju provinces respectively and the overall prevalence in all the farms was 43.2% by means of acridine orange stain. The parasites that were observed in the peripheral blood of calves was shown surely by the western immunoblot to the characteristic 34KD antigen among the proteins of T sergenti (Korean Isolate). And the antibody of the neonatal calves reacted at the very highest titer(1 : >2,520). These data highlight the significance of T sergenti in the neonatal calf disease in Korea.

• 대한수의학회지
• /
• 제34권2호
• /
• pp.417-419
• /
• 1994

최근 우리나라는 anaplasmosis 유행 지역으로부터 사슴을 비롯한 반추수를 수입하고 있다. 이처럼 리켓치아성 질환을 비롯한 원충성 질병의 국내 유입 기회가 높아지고 있다. 그러나 사슴에서의 anaplasmosis에 대한 연구보고 예는 접할 수 없었다. 1993년 전라북도의 한 사슴 목장에서는 호주 등지로부터 수입한 250 여두의 사슴에서 빈혈을 수반한 심한 쇄약증세를 나타내는 20마리 사슴의 혈액도말 표본을 Giemsa stain 과 acridine orange stain 방법으로 진단하였던 바, 이 중 8 마리에서 Anaplasma spp가 관찰되었기에 anaplasmosis에 의한 경제적 손실을 최소화하는데 일익을 도모하고자 사슴에서의 anaplasmosis 발병을 보고하는 바이다.

• 한국치위생학회지
• /
• 제14권2호
• /
• pp.287-292
• /
• 2014

Objectives : The purpose of the study is to examine the cell growth effect and autophagy effect of Scutellariae Radix by ethanol extract in SCC 25 cells. Methods : Cell growth inhibitory effect and autophagy induced by Scutellariae Radix were confirmed by WST-1 assay, monodansylcadaverine(MDC) stain, and flow cytometry by acridine orange(AO) stain. Results : The Scutellariae Radix treatment decreased the cell proliferation in a dose and time dependent manner. Scutellariae Radix has anticancer effects that autophagic vacuoles were apparent by MDC and AO staining in SCC 25 cells. Conclusions : Scutellariae Radix showed anticancer activity against SCC 25 cells via autophagy. The data provided the possibility that Scutellariae Radix may potentially contribute to oral cancer treatment.

• The Korean Journal of Physiology and Pharmacology
• /
• 제19권6호
• /
• pp.507-514
• /
• 2015

Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells.

• Parasites, Hosts and Diseases
• /
• 제26권4호
• /
• pp.229-238
• /
• 1988

HL-60세포에서 Toxoplasma gondii의 in vitro배양과 HL-60세포를 DMSO로 처리하여 과립세포로 분화시킨 세포에서 Toxoplasma에 대한 세포매개성 면역 기능을 검토하였다. 먼저, HL-60 세포를 여러 농도의 DMSO로 처리하였는데, 1.3%(V/V)로 3일간 처리하였을 때, HL-60 세포 의 적정 분화가 이루어졌다. 분화의 정도는 형태적, 생리적, 및 기능적 관점에서 검사되었는데, DMSO를 처리한 경우, 3H-thymidine의 흡입이 감소하는 것으로 보아 DNA 합성이 억제됨을 알 수 있었으며, 기능적으로는 주화성 물질인 FMLP에 대해 이동하는 성질을 보였으며, 형태적으로는 핵1세포질의 비가 큰 promyelocyte에서 작은 비를 갖는 과립 세포로 변화하여 분화를 입증하였다. 이후, HL-60 세포나 DMSO로 분화를 유도한 HL-60 세포와 Toxoplasma를 같이 배양하면서 이들의 관계를 관찰하였다. Lysosome에 선택적으로 흡입되는 형광 물질(acridine orange)로 전처리한 표본은 형광현미경하에 서 관찰하였으며, 다른 표본은 Giemsa로 염색하여 광학 현미경하에서 관찰하여 비교하였다. HL-60 세포에서는 72시간의 배양으로 Toxoplasma가 세포질내에서 증식하여 rosette를 형성하였으며, DMSO로 분화시킨 HL-60 세 포에서는 배양 초기 1시간째에 phagocytosis가 일어났으며 이후 세포내 소화가 이루어져 72시간째에는 lysosome이 원상태로 되돌아오는 것이 관찰되었다. 이상의 결과들로 볼 때, Toxoplnsma의 숙주 세포내에서의 기생 혹은 면역 세포에 의한 감수성에 phagosome 과 Iysosome의 융합이 결정적인 인자임을 알 수 있었으며, 아울러 HL-60 세포에서의 Toxoplasma의 증식 가능성과 DMSO로 분화시킨 HL-60 세포의 Toxoplasma 파괴 효과가 원충 기생충과 숙주의 상호 관계를 규명하는 좋은 모델임을 제시하였다.

• Tuberculosis and Respiratory Diseases
• /
• 제69권2호
• /
• pp.81-94
• /
• 2010

Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.