• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3471-3476
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• 2012

Background and Objective: Histone deacetylase (HDAC) inhibitors represent a promising class of potential anticancer agents for treatment of human malignancies. In this study, we investigated the effect of trichostatin A (TSA), one such HDAC inhibitor, in combination with docetaxel (TXT), a cytotoxic chemotherapy agent or erlotinib, a novel molecular target therapy drug, on lung cancer A549 cells. Methods: A549 cells were treated with TXT, erlotinib alone or in combination with TSA, respectively. Cell viability, apoptosis, and cell cycle distribution were evaluated using MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assay, Hochst33258 staining and flow cytometry. Moreover, immunofluorescent staining and Western blot analysis were employed to examine alterations of ${\alpha}$-tubulin, heat shock protein 90 (hsp90), epidermal growth factor receptor (EGFR), and caspase-3 in response to the different exogenous stimuli. Results: Compared with single-agent treatment, co-treatment of A549 cells with TSA/TXT or TSA/erlotinib synergistically inhibited cell proliferation, induced apoptosis, and caused cell cycle delay at the $G_2/M$ transition. Treatment with TSA/TXT or TSA/erlotinib led to a significant increase of cleaved caspase-3 expression, also resulting in elevated acetylation of ${\alpha}$-tubulin or hsp90 and decreased expression of EGFR, which was negatively associated with the level of acetylated hsp90. Conclusions: Synergistic anti-tumor effects are observed between TXT or erlotinib and TSA on lung cancer cells. Such combinations may provide a more effective strategy for treating human lung cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.1971-1975
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• 2012

Overexpression of several aquaporins (AQPs) has been reported in different types of human cancer but their role in carcinogenesis, for example in the cervix, have yet to be clearly defined. In this study, expression of AQPs in cervical carcinomawas investigated by real-time PCR, immunofluorescent and immunohistochemical assays and evaluated for correlations with clinicopathologic variables. AQP1, 3, 8 exhibited differential expression in cervical carcinoma, corresponding CIN and mild cervicitis. AQP1 was predominantly localized in the microvascular endothelial cell in the stroma of mild cervicitis, CIN and cervical carcinoma. AQP3 and AQP8 were localized in the membrane of normal squamous epithelium and carcinoma cells, local signals being more common than diffuse staining. AQP1 and AQP3 expression was remarkably stronger in cervical cancer than in mild cervicitis and CIN2-3 (P<0.05). AQP8 expression was highest in CIN2-3 (91.7%), but levels in cervical carcinoma were also higher than in mild cervicitis. AQP1, AQP3, AQP8 expression significantly increased in advanced stage, deeper infiltration, metastatic lymph nodes and larger tumor volume (P<0.05). Our findings showed that AQPs might play important roles in cervical carcinogenesis and tumour progression in Uygur women.

• Biomedical Science Letters
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• v.13 no.4
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• pp.273-278
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• 2007

Stem cells have been the subject of increasing scientific interest because of their utility in numerous biomedical applications. Stem cells are capable of renewing themselves; that is, they can be continuously cultured in an undifferentiated state, giving rise to more specialized cells of the human body. Therefore, stem cells are an important new tools for developing unique, in vitro model systems to test drugs and chemicals and a potential to predict or anticipate toxicity in humans. In the present study, in vitro cultured F3 immortalized human neural stem cell line and in vivo adult Sprague Dawley rats was used to evaluate the cytotoxicity of anticancer drug paclitaxel. In vitro apoptotic activity of paclitaxel was evaluated in F3 cell line by a MTT assay and DAPI test. The cell death was induced with the treatment of 20 nM paclitaxel and chromatin degradation was detected by DAPI staining, which was analyzed by fluorescent microscope. In vivo studies, we also observed nestin immunoreactivity on subventricular zone, which is stem cell rich region in the adult brain of the SD rat. Immunofluorescent staining result shows that pixel intensities of nestin were decreased in a dose dependent manner. These results suggest that paclitaxel is able to induce cytotoxic activity both in F3 neural stem cell line and neural stem cell in SD rat brain.

• BMB Reports
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• v.41 no.10
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• pp.745-750
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• 2008

Selenium, an essential trace element possessing anti-carcinogenic properties, can induce apoptosis in cancer cells. We have previously shown that sodium selenite can induce apoptosis by activating the mitochondrial apoptosis pathway in NB4 cells. However, the detailed mechanism remains unclear. Presently, we demonstrate that p53 contributes to apoptosis by directing signaling at the mitochondria. Immunofluorescent and Western blot procedures revealed selenite-induced p53 translocation to mitochondria. Inhibition of p53 blocked accumulation of reactive oxygen species (ROS) and loss of mitochondrial membrane potential, suggesting that mitochondrial p53 acts as an upstream signal of ROS and activates the mitochondrial apoptosis pathway. Selenite also disrupted cellular calcium ion homeostasis in a ROS-dependent manner and increased mitochondrial calcium ion concentration. p38 kinase mediated phosphorylation and mitochondrial translocation of p53. Taken together, these results indicate that p53 involves selenite-induced NB4 cell apoptosis by translocation to mitochondria and activation mitochondrial apoptosis pathway in a transcription-independent manner.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.11-18
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• 2000

Since $Hantavax^{TM}$, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with $Hantavax^{TM}$. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees at one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result $Hantavax^{TM}$ has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

• Korean Journal of Veterinary Research
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• v.40 no.1
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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.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.289-293
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• 2001

Incidence of infectious viruses is ensuing throughout the world and threatening the health of children as well as adults. The outbreaks of viral diseases of alimentary tract in Pusan from 1998 to 2000 were detected. Viruses were isolated from stool specimens, cerebrospinal fluid and throat swabs from suspicious patients and confirmed by cell culture, latex agglutination test, indirect immunofluorescent test and electron microscopic observation. The average isolation rate was 12.5% from the suspected specimens. From this work, 2 cases of enteric adenoviruses, 23 cases of echovirus, 31 cases of coxsackivirus 36 cases of rotavirus, 45 cases of SRSV, and 7 cases of poliovirus were detected. The major serotypes of coxsackievirus were B2, B3, B4, B6 and echovirus of serotypes 6, 9, 11, 25, and 30 were examined. Two cases of enteric adenovirus type 41 were also confirmed. The incidence of SRSV was mostly concentrated between December through following March, April through October with echovirus and coxsackievirus, and January through April with rotavirus, respectively. Electron micrograph of negative-stained viruses showed typical appearance with 30-80 nm in diameter.

• Journal of fish pathology
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• v.3 no.1
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• pp.11-19
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• 1990

Yellow tail (Seriola quinqueradiata) infected by Pasteurella piscicida have been occurred to mass mortality without showing apparent surface lesions in cage culture farms. In this case, it is necessary to consider countermeasure by rapid diagnosis of infected fish. The purpose of the present study was to investigate usefulness of the direct fluorescent antibody technique(FAT) for rapid diagnosis of pseudotuberclosis of cultured yellowtail caused by P. piscicida. Antibody produced by inoculating rabbit with formalin killed pseudotuberclosis bacteria antigen(strain KNP-2). Immunoglobulin-G(IgG) was purified from antisera by DEAE-cellulose column chromatography and conjugate with fluorescein isothiocyanate. Fluorescein-labeled antisera was purified by sephadex G-25 gel column chromatography. The fluorescein/protein molar ratio of labeled antisera was determined as 8.8-9.5. Diagnosis of cultured yellowtail was examined in cage culture farms which located in Tongyung, kyungnam from July to October 1990. The causative bacteria of pseudotuberclosis could be detected within two hours after the specimens were transferred to the laboratory for FAT, and it showed that FAT could be adapted as a rapid and accurate diagnostic method of pseudotuberclosis in yellowtail.

• The Journal of the Korean Society for Microbiology
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• v.19 no.1
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• pp.1-10
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• 1984

The occurrence of Hemorrhagic fever with renal syndrome was noted for the first time among the United Nations troops in Korea in 1951. In 1976, Lee and Lee demonstrated for the first time an antigen in the lungs of a striped field mouse, Apodemus agrarius, which gave specific immunofluorescent reactions with sera from patients convalescent from Korean hemorrhagic feve (KHF). The natural reservoir host of KHF is Apodemus agrarius coreae mice in rural endemic areas in Korea. Clinical manifestations of acute illness do not occur in infected Apodemus agrarius. Infected rodents excrete large amounts of virus in saliva, in urine and in feces for a long period of time. In this study, the tissues of Apodemus agrarius, Wistar rat and Balb/C mouse were inoculated with Hantaan virus and subsequently observed for any histopathologic findings. 1. In lungs, infiltration of lymphocytes in alveolar septa and peribronchial region and thickening of and giant cell formation in alveolar septa were observed. 2. In kidney, vascular congestion, interstitial hemorrhage in corticomedullary junction and partial microscopic hemorrhage in urinary space were found. 3. In spleens, vascular congestion, old and recent hemorrhage and multinucleated giant cell formation were seen.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.628-634
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• 2002

A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.