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

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*

AIMTo study the alkaloid constituents of Huperzia serrata (Thunb.) Trev..

METHODSChromatographic methods were used for the isolation and purification. Structure was elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSAn alkaloid constituent was isolated from H. serrata (Thumb.) Trev..

CONCLUSIONThe compound was found to be a novel phlegmariurine type alkaloid, named 8 beta-hydroxy phlegmariurine B.

AIM: To study the minor diterpenes from the soft coral Sinularia depressa METHOD: The chemical constituents were isolated and purified by various chromatographic techniques, and the chemical structures, including absolute configuration, were established on the basis of detailed analysis of spectroscopic data and by literature comparison with the data of related known compounds. RESULTS: A new casbane-type diterpene, 2-epi-10-hydroxydepressin (1), was isolated and identified. CONCLUSION Compound 1 is a new casbane-type diterpene.
Animals ; Anthozoa ; chemistry ; Diterpenes ; isolation & purification ; Heterocyclic Compounds, 3-Ring ; Hexamethonium ; analysis ; Spectrum Analysis

OBJECTIVEThis study was purposed to investigate the effect of Akt kinase inhibitor MK2206 on proliferation and apoptosis of U937 cells and RS4;11 cells, and to explore its possible mechanism.

METHODSU937 and RS4;11 cells were cultured with different concentrations of MK2206 for 24 h and 48 h, and cell growth curve was analyzed by CCK-8; cell apoptosis was analyzed by Annexin V/7-AAD double labeling; cell cycle changes were analyzed by flow cytometry. The BAX, BCL-2, XIAP, CDK1, caspase-3 mRNA expressions were determined by real time PCR.

RESULTSMK2206 significantly inhibited the growth of U937 and RS4;11 cells in a time-and dose-dependent manner, and the IC50 values of U937 cells for 24 h and 48 h were (0.48±0.15) µmol/L and (0.09±0.01) µmol/L respectively, while IC50 values of RS4;11 cells for 24 h and 48 h were (0.91±0.02) µmol/L and (0.68±0.11) µmol/L respectively. U937 were cultured with 0.5 µmol/L MK2206 and RS4;11 cells were cultured with 1.0 µmol/L MK2206 for 24 h and 48 h, and the both apoptosis rates were higher for 24 h or 48 h than that in control group (P<0.05), meanwhile the apoptosis rates for 48 h were higher than 24 h. The results of cell cycle detection showed that the both cells were arrested in G2/M phase compared with control group. The real time PCR assay revealed that the expressions of BAX, caspase-3 mRNA in cells treated with MK2206 were increased, while BCL-2, XIAP, CDK1 were reduced compared with control group.

CONCLUSIONMK2206 can inhibit proliferation and induce apoptosis of U937 and RS4;11 cells, and the both cells are arrested in G2/M phase. The mechanism of promoting apoptosis may be related with up-regulating BAX, caspase-3 and down-regulating BCL-2, XIAP, meanwhile the cell cycle arrested in G2/M phase may be associated with down-regulating CDK1.

Tadalafil is an effective drug in treating erectile dysfunction (ED), and its clinical efficacy has been confirmed by a great many researches. Tadalafil is distinguished from sildenafil and vardenafil by its prolonged action lasting 36 hours for a sigle dose, compared with about 4 hours for sildenafil. Furthermore, this drug is effective in improving the erectile function of ED patients including those with various comorbid conditions. Tadalafil can help ED patients to regain morning erection and recover the confidence as a man. More and more ED patients choose tadalafil as the first line therapy because of its long efficacy and its conformability to the therapeutic requirement by restoring ED patients to normal, natural and pleasurable sexual life.
Carbolines ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Tadalafil

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 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*

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