In order to perform standardization in analytic methods of cryopreserved human semen, to investigate the differences of resistance to cryoinjury, to define the stage of critical cryoinjury during cryopreservation and to evaluate the quality change after thawing by time interval, the reviability of 30 normal and 30 abnormal semen were evaluated by the supravital stainings of spermatozoa using acridine orange and eosin yellow and the motility assay simultaneously according to the stages of freezing-thawing and the time interval after thawing. The results were summarized as follows: 1. Vitality was estimated higher than motility at all specimens and the gap between two became greater as motility decreased. 2. Reviability of abnormal semen was estimated lower than that of normal semen(p<0.05). 3. The critical cryoinjury to spermatozoa was noticed at the stage of freezing from 4 degrees C to -10 degrees C (p<0.05). 4. The significant decrease in quality of normal cryopreserved semen was noticed between 30 to 60 min. after thawing (p<0.05). These results suggest that the cryoinjury to human semen is different in nature, therefore it is advisable that the quality of cryopreserved human semen should be evaluated by vitality and motility assay simultaneously. And the resistance of abnormal semen to cryopreservation is so low that it would be difficult to be used clinically with satisfaction. Moreover the laboratory studies should be concentrated on freezing method to achieve better reviability and it is desirable in practice that post-thaw semen should be used within 30 min, after thawing.
Acridine Orange ; Cryopreservation ; Eosine Yellowish-(YS) ; Freezing ; Humans* ; Semen ; Spermatozoa*

BACKGROUND: In South Korea, indigenous malaria has been reappeared since 1993 and more than 350 cases diagnosed in 1996. For the diagnosis of malaria the classic methods such as thin and thick blood smears with Giemsa or Wright stain has been routinely used. Since recently fluorochrome staining has been shown to be more sensitive, easy to do, and less time-consuming, we applied the new method, Acridine orange stain, for diagnosis of clinically suspected cases. METHODS: Thin and thick blood smears were prepared from civilian patients of Kyunggi Province (n=20) and Republic Of Korea army patients pre- (n=67) and post-treatment (n=13) of malaria. The slides were fixed by methanol and stained by either Giemsa or Acridine orange solution (10-50 g/mL). For comparison, an expert on malaria diagnosis examined them by light and fluorescent microscope, respectively. RESULT: Acridine orange stain was found to be a rapid technique, and as sensitive (83%) as thick smears (83%) for diagnosis of malaria. The detection limit of acridine orange stain was 23.5 parasites/ul of blood. The staining time was much shorter (30 sec) than that of Giemsa stain (30-60min). CONCLUSION: Acridine orange stain is evaluated as a simple, rapid, and sensitive method for malaria diagnosis compared with Giemsa stain.
Acridine Orange* ; Azure Stains ; Diagnosis* ; Gyeonggi-do ; Humans ; Korea ; Limit of Detection ; Malaria* ; Methanol ; Republic of Korea

Purpose: Colonization of denture soft lining materials by Candida albicans can result in clinical problem, and deterioration of the materials. This study aimed to compare the retention and penetration of C. albicans into four denture soft lining materials commonly used. Materials and methods: Four denture soft lining materials(Coe-comfort., Coe-soft., GC soft liner., and Tissue conditioner.) discs were prepared to glass slide and dental stone. Adherence of yeast to surfaces was monitored after one hour incubation of standardized washed cell suspension with test disc surfaces. Adherent cells stained with acridine orange were counted fluorescence microscopy. Penetration of yeast into materials bonded with acrylic resin after 1, 2, 3, 4, 5, 6 and 7 days incubation was observed through sections stained using acridine orange and estimated to quantitative analysis using radioisotope. Results: There was statistical significance in cell numbers between smooth and rough surfaces(p<0.05). Higher numbers of cells were observed on rough surfaces. There was statistical significance in adherent cell numbers into smooth and rough surfaces individually(p<0.05). According to the increase of incubation periods, the cells penetrated into denture soft lining materials were shown to increase. The differences among all kinds of soft liner were statistically significant(p<0.05),and the largest number of cells penetrated into soft liners was observed in the Coe-soft. Conclusion: Initial adherence and penetration of yeast into denture soft lining materials has been influenced by surface roughness and chemical composition of them. The selection of appropriate materials and their fabrication may promote clinical performance.
Acridine Orange ; Candida albicans* ; Candida* ; Cell Count ; Colon ; Dentures* ; Glass ; Microscopy, Fluorescence ; Stomatitis, Denture ; Yeasts

We investigated the role of autophagy in SNUC5/5-FUR, 5-fluorouracil (5-FU) resistant SNUC5 colon cancer cells. SNUC5/5-FUR cells exhibited low level of autophagy, as determined by light microscopy, confocal microscopy, and flow cytometry following acridine orange staining, and the decreased level of GFP-LC3 puncta. In addition, expression of critical autophagic proteins such as Atg5, Beclin-1 and LC3-II and autophagic flux was diminished in SNUC5/5-FUR cells. Whereas production of reactive oxygen species (ROS) was significantly elevated in SNUC5/5-FUR cells, treatment with the ROS inhibitor N-acetyl cysteine further reduced the level of autophagy. Taken together, these results indicate that decreased autophagy is linked to 5-FU resistance in SNUC5 colon cancer cells.
Acridine Orange ; Autophagy* ; Colon* ; Colonic Neoplasms* ; Cysteine ; Flow Cytometry ; Fluorouracil* ; Microscopy ; Microscopy, Confocal ; Reactive Oxygen Species

The Ziehl-Neelson's AFB staining method was mainly used for the AFB observation of the mycobacteria. However, this method has several issues of false negative results, and hence a comparative experiment of the Ziehl-Neelson's AFB staining and the fluorescence staining method was done to remedy this problem. As the fluorescence staining method brightly highlights the AFB in a dark field, and also as it is observed with the lower power objective, it is a method that can better the observation and shorten the time of observation as well. The fluorescence staining method that was used in this experiment did a comparative analysis of the Auramine O-Rhodamine B and the Acridine Orange. The results showed that although the Auramine O-Rhodamine B allows easier observation of the AFB with a high fluorescence expression rate for the multibacillary leprosy sample, the darkness on the periphery makes it hard to observe anything else, while also making it hard to observe the cell changes and paucibacillary leprosy of the AFB. However, the Acridine Orange staining method highlights the cells in dark green and changes the color of the AFB from bright red to orange making it easier to observe bacilli. The results of the study show that the Acridine Orange method is superior to the Auramine O-Rhodamine B method in detecting acid fast bacilli in specimen.
Acridine Orange ; Benzophenoneidum ; Citrus sinensis ; Darkness ; Fluorescence ; Leprosy, Multibacillary ; Leprosy, Paucibacillary ; Mycobacterium ; Mycobacterium leprae

We assessed the comparatice sensitivity and specificity of acridine orange and Gram stains of corneal scrapings from 29 cases of presumed infectious keratitis. The corneal scrapings were placed on two clear glass slides and allowed to air dry. The slides were fixed in 95% methanol for 2 minutes and each slide was stained by Gram and acridine orange stains respectively. Culture was done with another corneal scraping. Cultures were positive in 19 of 29 cases(66%). Among them 14 cases (74%) were positive in Gram stain and 18 cases (95%) were positive in acridine orange stain. So the sensitivity of acridine orange was greater than that of Gram stain. In the specificity, Gram stain (80%) was better than acridine orange stain(50%). This study demonstrates acridine orange stain as well as gram stain is useful as a simple, rapid screening test for the diagnois of suspected infectious keratitis.
Acridine Orange* ; Coloring Agents* ; Diagnosis* ; Glass ; Keratitis* ; Mass Screening ; Methanol ; Sensitivity and Specificity

Several observation suggest that the antimelanocyte autoantibodies could play a role in melanocyte destruction. Some experiments indicate that melanocyte antibodies from patients with vitiligo can kill melanocyte in vitro. In these experiments, we demonstrated that vitiligo patient's sera containing antimelanocyte antibodies can lyse cultured human melanocytes by complement activation. Melanocyte cytotoxicity was measured using the ethidium bromide/ acridine orange viability assay. Significant melanocyte cytotoxicity was seen in sera from patients with both active and inactive vitiligo(p<0.01). Melanocyte cytotoxicity measured with complement-mediated cytotoxicity decreased after systemic steroid treatment(p<0.05) ; however melanocyte cytotoxicity showed no significant change with systemic PUVA therapy.
Acridine Orange ; Antibodies ; Autoantibodies* ; Complement Activation ; Ethidium ; Humans ; Melanocytes ; PUVA Therapy ; Vitiligo*

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles

We report a case of vivax malaria that showed ruptured form of merozoite only in the peripheral blood. A 28-year old man was admitted to Korea University hospital because of irregular high fever, chill and abdominal pain. The peripheral blood smear, showed only small merozoites which seemed to have been recently released from schizonts and destroyed remnants form of schizonts and did not show any forms of malaria parasite such as ring forms, mature trophozoites, schizonts, and gametocytes. On acridine orange fluorochrome stain, we could not find any suspected forms of malaria. However, we detected parasite LDH which is specific to Plasmodium vivax. Malaria treatment was done to the patient, and he is now under follow up in local hospital.
Abdominal Pain ; Acridine Orange ; Adult ; Fever ; Follow-Up Studies ; Humans ; Korea ; Malaria ; Malaria, Vivax* ; Merozoites* ; Parasites ; Plasmodium vivax ; Schizonts* ; Trophozoites