• Molecules and Cells
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• 제39권10호
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• pp.742-749
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• 2016

The cancer chemo-preventive effects of equol have been demonstrated for a wide variety of experimental tumours. In a previous study, we found that equol inhibited proliferation and induced apoptotic death of human gastric cancer MGC-803 cells. However, the mechanisms underlying equol-mediated apoptosis have not been well understood. In the present study, the dual AO (acridine orange)/EB (ethidium bromide) fluorescent assay, the comet assay, MTS, western blotting and flow cytometric assays were performed to further investigate the pro-apoptotic effect of equol and its associated mechanisms in MGC-803 cells. The results demonstrated that equol induced an apoptotic nuclear morphology revealed by AO/EB staining, the presence of a comet tail, the cleavage of caspase-3 and PARP and the depletion of cIAP1, indicating its pro-apoptotic effect. In addition, equol-induced apoptosis involves the mitochondria-dependent cell-death pathway, evidenced by the depolarization of the mitochondrial membrane potential, the cleavage of caspase-9 and the depletion of Bcl-xL and full-length Bid. Moreover, treating MGC-803 cells with equol induced the sustained activation of extracellular signal-regulated kinase (ERK), and inhibiting ERK by U0126, a MEK/ERK pathway inhibitor, significantly attenuated the equol-induced cell apoptosis. These results suggest that equol induces mitochondria-dependent apoptosis in human gastric cancer MGC-803 cells via the sustained activation of the ERK1/2 pathway. Therefore, equol may be a novel candidate for the chemoprevention and therapy of gastric cancer.

• 대한미생물학회지
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• 제21권1호
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• pp.33-45
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• 1986

On hundred and forty stains of shigella cultures isolated from the twelve hygiene laboratories of cities and provincial general hospital laboratories in 1983 were tested for their resistance to thirteen antimicrobial drugs and their R-plasmid transfer. Antimicrobial drugs were used amikacin, ampicillin, cephalothin, chloramphenicol, gentamicin, kanamycin, nalidixic acid, rifampicin, streptamycin, tetracycline, tobramycin, cefoperazone and piperacillin. All strains were resistant to one or more of thirteen antimicrobial drugs but 94.3% were susceptible to amikacin, gentamicin and tobramycin of total isolated. The most strains commonly found resistance was to chloramphenicol (94%) followed by streptamycin (93%), tetracyline (92%) piperacillin (90%) ampicillin (83%), cefoperazone (42%), nalidixic acid (14%), cephalothin (17%), rifampicin (22%) and kanamycin (6%), sixty percent of strains among 140 were resistance to ampicillin, chloramphenicol, streptomycin, tetracycline at the same time. The transfer of drug resistance by conjugation was tested and ninety four strains (94.3%) were resistant to one or more drugs were found to transfer their drug resistance of E. coli. percentage of transfer frequency by conjugation was one strains (54%), the transfer frequency of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to recipients. Percentage of plasmid curing after the treatment of acriflavine, acridine orange was about 8%. Among strains cured two strains were tested compare original strains with them in biochemical properties in arginine dihydrolase and arabinose fermentation reaction. It was found to growth curves of No.2 shigella flexneri, serotype 1b, and its derivatives cured with acriflavine in $M{\ddot{u}}ller$ Hinton broth medium (pH 7.4, $38^{\circ}C$) by temperature Gradient Biophoto Recorder TN-1120 (Tokyo, Japan).

• 대한수의학회지
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• 제35권1호
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• pp.137-142
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• 1995

Anaplasmosis is a tick-borne disease of large and small ruminants, causing losses through mortality, abortion, weight loss and reduced milk production. In one dairy farm, for example, 250 of a total of 800 imported goats were diagnosed with a mysterious type of anemia during the summer and autumn of 1992. The etiologic agent was identified as Anaplasma spp by acridine orange and ultrastructure by electron microscopy. In order to monitor variations in blood biochemical and hematological parameters associated with the disease, blood samples were collected by jugular venipuncture from 50 goats at 3 month intervals between the period of February and October, 1993. The levels of RBCs, HB and HCT decreased from $18.48{\pm}1.96$ to $13.47{\pm}2.48X10^6/mm^3$, $12.25{\pm}1.41$ to $9.54{\pm}1.77g/dl$, and $43.09{\pm}4.75$ to $30.93{\pm}5.78%$, respectively. The values of MCH(Mean corpuscular hemoglobin), MCHC(Mean corpuscular hemoglobin concentration) and PLT(Platelet) were elevated from $6.58{\pm}0.30$ to $7.05{\pm}0.47pg$, $28.40{\pm}1.20$ to $30.82{\pm}1.85g/dl$ and $1688.34{\pm}750$ to $2046.82{\pm}783X10^3/mm^3$, respectively. Percent parasitized erythrocytes(PPE) increased from $0.61{\pm}0.5$ to $2.22{\pm}1.9%$, clinical biochemical parameters aspartate aminotransferase and alanine aminotransferase were $66.64{\pm}23.1K.U$ and $14.90{\pm}6.59K.U$, respectively and persisted at high levels throughout the observation period. The level of albumin(2.46)0.52 g/dl) was decreased corresponding to an elevated globulin and a reduced albumin/globulin ratio in October as compared with the values in February. It is concluded that caprine anaplasmosis may be an important cause of anemia and hepatic malfunction in goats.

• Tuberculosis and Respiratory Diseases
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• 제72권4호
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• pp.343-351
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• 2012

Background: The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling axis has emerged as a novel target for cancer therapy. Agents that inhibit this pathway are currently under development for lung cancer treatment. In the present study, we have tested whether dual inhibition of PI3K/Akt/mTOR signaling can lead to enahnced antitumor effects. We have also examined the role of autophagy during this process. Methods: We analyzed the combination effect of the mTOR inhibitor, temsirolimus, and the Akt inhibitor, GSK690693, on the survival of NCI-H460 and A549 non-small cell lung cancer cells. Cell proliferation was determined by MTT assay and apoptosis induction was evaluated by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Autophagy induction was also evaluated by acridine orange staining. Changes of apoptosis or autophagy-related proteins were evaluated by western blot analysis. Results: Combination treatment with temsirolimus and GSK690693 caused synergistically increased cell death in NCI-H460 and A549 cells. This was attributable to increased induction of apoptosis. Caspase 3 activation and poly(ADP-ribose) polymerase cleavage accompanied these findings. Autophagy also increased and inhibition of autophagy resulted in increased cell death, suggesting its cytoprotective role during this process. Conclusion: Taken together, our results suggest that the combination of temsirolimus and GSK690693 could be a novel strategy for lung cancer therapy. Inhibition of autophagy could also be a promising method of enhancing the combination effect of these drugs.

• Clinical and Experimental Reproductive Medicine
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• 제43권2호
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• pp.90-96
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• 2016

Objective: Diabetes mellitus (DM) is known to cause many systemic complications as well as male infertility. Astaxanthin (ASTX) is a powerful antioxidant that is involved in a variety of biologically active processes, including those with anti-diabetes effects. The present study investigates the effect of ASTX on the spermatozoa function in streptozotocin (STZ)-induced diabetic rats. Methods: We divided 30 adult rats into three groups (10 rats per group), with a control group that received corn oil mixed with chow. DM was induced by intra-peritoneal injection of STZ. Eight weeks after the STZ injection, half of the diabetic animals were used as diabetic controls, and the rest were treated with ASTX for 56 days. Then the parameters and chromatin integrity of the epididymal sperm were analyzed using chromomycin A3, toluidine blue (TB), and acridine orange (AO) staining. Results: The count, viability, and motility of the epididymal sperm were decreased significantly in the STZ group in comparison with the control group (count and viability, p<0.001; motility, p<0.01). ASTX increased normal morphology and viable spermatozoa compared to the STZ group (morphology, p=0.001; viability, p<0.05). The percentage of abnormal chromatins in TB and AO staining was higher in the STZ group compared to the control group (p<0.001). The mean percentage of TB and AO positive spermatozoa in STZ rats was significantly lower in the STZ+ASTX group (TB, p=0.001; AO, p<0.05). Conclusion: This study observed that in vivo ASTX treatment partially attenuates some detrimental effect of diabetes. Conversely, ASTX improved sperm viability, normal morphology, and DNA integrity.

• 미생물학회지
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• 제33권1호
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• pp.7-14
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• 1997

성장인자가 포함되지 않은 무기염 액체배지(ISP-4)에서 Streptomyces coelicolor A3(2)의 포자가 발아할 때, 성장인자의 주기적인 요구성이(양 등,1993_ 단백질과 RNA 합성과 어떤 관계를 가지고 있는지를 규명하고자 하ㅣ였다. 발아는 10시간 정도 걸렸으며 이때 성장인자의 요구성이 2시간 주기로 반복되는 것을 재입증하였다. 포자의 크기는 시간에 따라 증가하였으나, 포자수는 표준 평판계수법에서 감소하였다. 포자 집단은 생리적으로 살아있거나, 휴면 중인 포자와 죽은 포자로 구분될 수 있었다. 이러한 발아과정에서 포자를 acridine orange(AO)와 iodonitrotetrazolium chloride(INT)로 염색하여 형광현미경으로 관찰하였을 때 RNA와 단백질도 일정한 주기를 갖고 합성되었으며, 이 주기성은 성장인자의 요구 주기와 거의 일치하였다. 이로써 발아 초기에 포자 집단은 성장인자 중에서 특히 단백질 합성과 관련된 물질인 아미노산이 주기적으로 요구되고 있음을 토론하였다.

• International Journal of Oral Biology
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• 제38권3호
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• pp.93-100
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• 2013

Bile acids and synthetic bile acid derivatives induce apoptosis in various kinds of cancer cells and thus have anticancer properties. Recently, it has been suggested that autophagy may play an important role in cancer therapy. However, few data are available regarding the role of autophagy in oral cancers and there have been no reports of autophagic cell death in OSCCs (oral squamous cell carcinoma cells) induced by HS-1200, a synthetic bile acid derivative. We thus examine whether HS-1200 modulates autophagy in OSCCs. Our findings indicate that HS-1200 has anticancer effects in OSCCs, and we observed in these cells that autophagic vacuoles were visible by monodansylcadaverine (MDC)and acridine orange staining. When we analyzed HS-1200-treated OSCC cells for the presence of biochemical markers, we observed that this treatment directly affects the conversion of LC-3II, degradation of p62/SQSTM1 and full-length beclin-1, cleavage of ATG5-12 and the activation of caspase. An autophagy inhibitor suppressed HS-1200-induced cell death in OSCCs, confirming that autophagy acts as a pro-death signal in these cells. Furthermore, HS-1200 shows anticancer activity against OSCCs via both autophagy and apoptosis. Our current findings suggest that HS-1200 may potentially contribute to oral cancer treatment and thus provide useful information for the future development of a new therapeutic agent.

• International Journal of Oral Biology
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• 제42권1호
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• pp.17-23
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• 2017

Background: Periodontitis is generally a chronic disorder characterized by the breakdown of tooth-supporting tissues. P. gingivalis, a Gram-negative anaerobic rod, is one of the major pathogens associated with periodontitis. Frequently, P. gingivalis infection leads to cell death. However, the correlation between P. gingivalis-induced cell death and periodontal inflammation remains to be elucidated. Among cell deaths, the death of immune cells appears to play a significant role in inflammatory response. Thus, the aim of this study was to examine P. gingivalis-induced cell death, focusing on autophagy and apoptosis in THP-1 cells. Methods: Human acute monocytic leukemia cell line (THP-1) was used for all experiments. Autophagy induced by P. gingivalis in THP-1 cells was examined by Cyto ID staining. Intracellular autophagic vacuoles were observed by fluorescence microscopy using staining Acridine orange (AO); and 3-methyladenine (3-MA) was used to inhibit autophagy. Total cell death was measured by LDH assay. Cytokine production was measured by an ELISA method. Results: P. gingivalis induced autophagy in an MOI-dependent manner in THP-1 cells, but 3-MA treatment decreased autophagy and increased the apoptotic blebs. P. gingivalis infection did not increase apoptosis compared to the control cells, whereas inhibition of autophagy by 3-MA significantly increased apoptosis in P. gingivalis-infected THP-1 cells. Inhibition of autophagy by 3-MA also increased total cell deaths and inflammatory cytokine production, including $IL-1{\beta}$ and $TNF-{\alpha}$. Conclusion: P. gingivalis induced autophagy in THP-1 cells, but the inhibition of autophagy by 3-MA stimulated apoptosis, leading to increased cell deaths and pro-inflammatory cytokines production. Hence, the modulation of cell deaths may provide a mechanism to fight against invading microorganisms in host cells and could be a promising way to control inflammation.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.101-107
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• 2020

Objective: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. Methods: Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. Results: In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p< 0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p< 0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. Conclusion: This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.143-150
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• 2002

Fluorescent probes bound to DNA typically display nanosecond decay times and reveal only nanosecond motions. We extend the time range of measurable DNA dynamics using $[Ru(pby)_2(dppz)]^{2+}$ (bpy=2.2'-bipyridine, dppz=dipyrido[3,2-a2',3'-c]phenazine) (RuBD) which displays a mean lifetime near 90 ns. To test the usefulness of RuBD as a probe for diffusive processes in calf thymus DNA, we compared the efficiencies of fluorescence resonance energy transfer (FRET) using three donors which display lifetimes near 5 ns for acridine orange (AO), 22 ns for ethidum bromide (EB) and 92 ns for RuBD, with nile blue (NB) as the acceptor. The F rster distances for AO-NB, EB-NB and RuBD-NB donor-acceptor pairs were 42.3, 52.3, and $30.6{\;}{\AA}$, respectively. All three donors showed dramatic decreases in fluorescence intensities and more rapid intensity decays with increasing NB concentrations. The intensity decays of AO and EB in the presence of varying concentrations of NB were satisfactorily described by the one-dimensional FRET model without diffusion (Blumen and Manz, 1979). In the case of the long-lifetime donor RuBD, the experimental phase and modulation somewhat deviated from the recovered values computed from this model. The recovered NB concentrations and FRET efficiencies from the model were slightly larger than the expected values, however, the recovered and expected values did not show a significant difference. Thus, it is suggested that the lifetime of RuBD is too short to measure diffusive processes in calf thymus DNA.