• YAKHAK HOEJI
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• v.45 no.1
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• pp.64-70
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• 2001

Protein-polysaccharide fractions, PJ-3 and PJ-4, were prepared from mycelial culture filtrate of an insect-born fungus, Paecilomyces japonica DGUM 32001, and subjected to a flow cytometrical analysis for their vivo antitumor and immunomodulating activity in ICR mice. When i.p. injected once daily for semen days at 100 mg/kg, PJ-4 exerted a strong antitumor activity showing the growth inhibition ratio of 85.1% against i.p. implanted sarcoma 180 cells, while PJ-3 showed only a weak activity. Moreover, PJ-4 signiscantly increased the expression level of CD25 (IL-2R $\alpha$-chain) as well as forward scatter (FSC) values of splenic CD8$^{8}$ T cells. It is also noteworthy that PJ-4 strongly induced the peritoneal exudate cells in the same experiment. In an in vitro study, PJ-4 slightly inhibited the growth of sarcoma 180 cells at the concentration of 50$\mu$g/ml or higher. These results strongly suggest that PJ-4 might exert its antitumor activity through immunostimulation as well as direct inhibitory activity on the tumor cells.

• Fisheries and Aquatic Sciences
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• v.26 no.7
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• pp.447-454
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• 2023

Fast somatic growth is important considerations for successful and competitive aquaculture industry. In rainbow trout reared in South Korea, triploid induction was used to suppress negative influence of reproductive maturation to body growth. However, the effects of triploidy are visible in both mature fish and developing juvenile fish. Thus, it is also important to explicate the effect of triploid induction on growth during the early-life stages of rainbow trout-alevins and fry. Rainbow trout fertilized eggs were subjected to triploid induction and polyploidy was checked by flow cytometry. Diploid and triploid alevins and fry were reared separately in tanks with constant flow of freshwater through flow-through water system and growth measurements were done from zero days after hatching (DAH 0) until DAH 134. The egg-yolk morphometrics of alevins-yolk length, yolk height, yolk volume and yolk weight-were statistically similar (p > 0.05) in both genotypes from DAH 0 to DAH 22. The total length, body height, and body weight of alevins and fry were statistically better (p > 0.05) in both genotypes until DAH 92 but thereafter, triploid had a significantly better growth performance (p < 0.05) over diploid fish until the completion of study at DAH 134. With that, triploid induction did not influence alevin yolk regions and body growth and fry somatic growth until around 3 months after hatching, but considerable growth enhancement was subsequently apparent.

• Biomedical Science Letters
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• v.1 no.1
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• pp.19-25
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• 1995

Characterization of immune cell subpopulations in the bovine was performed using a direct immunofluorescence technique adaptable for routine and repeated monitoring. This whole blood procedure is faster and requires less volume than conventional density gradient isolation methods. Low inter-and intra-animal variations were seen in hematology parameters and in CD4, CD8 and CD2 lymphocyte subtypes. CD4 values were 30% of lymphocytes in male and 32% in females. Thriteen percent were CD8 in males and 13% in females. CD4: CD8 ratios were approximately two in both sexes. Fiffty three percent were CD2 in males and 54% in females. The mean RBC counts of peripheral blood were 7.20$\times10^6/{mm}^3$ for male cattle and 6.36$\times10^6/{mm}^3$ for females. The mean WBC counts were 8.09$\times10^3/{mm}^3$ for males and 7.09$\times10^3/{mm}^3$ for females. The percent of lymphocytes(63-65%) was higher than the percent of neutrophils(17-18%), the percent of eosinophils(11-15%), te percent of monocytes(4-5%), and percent of basophiles(1%).

• Ocean and Polar Research
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• v.27 no.4
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• pp.397-417
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• 2005

Phytoplankton community structure and distribution pattern in the surface water around the Ieodo Ocean Research Station were investigated during seven cruises carried out from July, 2003 to October, 2004. Samples were analyzed using various tools including a microscope, flow cytometer, and HPLC. Satellite images were used to analyze spatio-temporal phytoplankton biomass distribution. SeaWiFS chlorophyll a (chl a) images showed that spring blooms occurred in April-May near the Ieodo Station, and these waters were under the influence of Changjiang Dilute Water during July-October. Also, during the July-October period, HPLC pigments data showed increasing zeaxanthin concentrations, a marker pigment of cyanobacteria whereas increasing concentrations of various other pigments such as fucoxanthin, peridinin, prasinoxanthia alloxanthin, 19'-hexanoyloxyfucoxanthin and chlorophyll b were noted during spring blooms. Such pigment marker data were consistent with picoplankton data analyzed by flow cytometer and nano-microplankton analyzed by microscope. The pigment-CHEMTAX method was used to drive the phytoplankton group apportioned chi a. Diatoms, chlorophytes, dinoflagellates, and cryptophytes comprised 25.8, 20.7, 15.9, and 14.1%, respectively, of the total chl a in May. Average cyanobacteria concentrations in July-October contributed 25.4% of the total concentration. This was the highest percent contribution and was followed by chlorophytes, diatoms, and prymnesiophytes. This study discusses results from various methods, similarities and differences in the results among those methods, and the application range of the results from different analytical methods. Also, the study reveals a detailed phytolpankton community structure in the waters around the Ieodo Station, and suggests future monitoring considerations in relation to cell morphology, ecology and diversity factors according to taxonomic groups.

• Clinical and Experimental Pediatrics
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• v.45 no.3
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• pp.302-310
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• 2002

Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells(PBMC) from three XLA families in Korea. Methods : Heparinized venous blood samples were collected from four XLA patients and additional family members in three unrelated XLA families. Mononuclear cells were separated from their blood and the intracellular Btk protein was characterized by a flow cytometry. The mutation analysis was performed using direct sequencing. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in the patients with XLA. We observed a novel deletion and two point mutations within introns(intron 1 and intron 18) resulting in alternative splicings. In XLA family 2, a 980 bp deletion(from intron 9+191 T to intron 10-215 C) including exon 10 was found in patient P2. He was the only sporadic case in this study, because his mother and brother showed a normal Btk expression by flow cytometry. Conclusion : These identified genetic alterations support the molecular heterogeneity of Btk gene in XLA disease. Additionally, by means of flow cytometric analysis, we diagnosed three hypogammaglobulinemia patients as XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease.

• Journal of Oral Medicine and Pain
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• v.34 no.3
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• pp.261-276
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• 2009

Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.200-205
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• 2004

Uterine leiomyoma is the most common tumor in the female genital tract. Although the tumor is benign, it is of paramount importance since it often causes profuse menstrual bleeding, pressure symptoms, and infertility. Nevertheless, the etiology and patholphysiology of this abnormality remain poorly understood. The traditional definitive treatment for uterine leiomyomas is hysterectomy and, even today, symptomatic leiomyomas are the leading cause of hysterectomy in Korea. Clearly, the development of a safe, effective, and nonsurgical method of treatment for leiomyoma would be of great benefit to many women. The present study was designed to investigate the effect of Rhubarb on apoptosis in uterine leiomyoma cells. Results demonstrate that Rhubarb inhibited cell growth in dose-dependent manner. Cell growth significantly decreased to 60% of control in the treatment of Rhubarb (300㎍/㎖). Associated with the decreased response, there was a concomitant and significant delay of subG1 8.32% above baseline in the treatment of Rhubarb (300㎍/㎖). The delay of subG1 showed a dose-dependent manner, as evidenced by the flow cytometry. The reduced cellular viability on exposure to Rhubarb may represent the induction of apoptosis, at least in part, as concomitantly evidenced by enhanced DNA fragmentation, PARP cleavage and caspase 9 and decreased pro-caspase 3. In addition, Rhubarb decreased clAP1 expression levels in dose-dependent manner. Talcen together, there results suggest that Rhubarb can produce a potent inhibition effect of apoptosis and implicate the delay of G1 phase in the cell cycle and pathways of caspase 3 and 9 in the mechanism underlying inhibitory apoptosis effect of Rhubarb.

• Journal of Acupuncture Research
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• v.19 no.1
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• pp.189-202
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• 2002

Objective : This study is designed to investigate the effects of bee venom and melittin on cell death in neuroblastoma cell line after pretreatment with NG(nerve growth inhibitory substance) Methods : It was evaluated by using MTT assay, morphological method, DNA fragmenation, flow cytometry, immunocytochemistry analysis, RT-PCR and Western blot. Results : The MTT assay demonstrated that neuroblastoma cell viability was significantly inhibited dose-dependently by treatment with bee venom and melittin after pretreatment with NG in comparison awith control. The morphological study and fow cytometry demonstrated that neuroblastoma cell showed apoptosis. DNA fragmenation showed DNA ladder below 1 Kbp. Immunocytochemistry assay demonstrated that Fos and MAPK were down-regulated. RT-PCR analysis demonstrated that Fos and MAPK was down-regulated. Western blot demonstrated that Fos and MAPK were down-regulated from $1{\mu}g/ml$ bee venom in neuroblastoma cell pretreated with NG. Conclusion : These result suggests that bee venom and melittin after NG treatment have significant anti-cancer effect and further study is needed in vivo.

• The Korean journal of internal medicine
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• v.34 no.1
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• pp.146-155
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• 2019

Background/Aims: Indoxyl sulfate (IS) is a uremic toxin and an important causative factor in the progression of chronic kidney disease. Recently, paricalcitol (19-nor-1,25-dihydroxyvitamin D2) was shown to exhibit protective effects in kidney injury. Here, we investigated the effects of paricalcitol treatment on IS-induced renal tubular injury. Methods: The fluorescent dye 2',7'-dichlorofluorescein diacetate was used to measure intracellular reactive oxygen species (ROS) following IS administration in human renal proximal tubular epithelial (HK-2) cells. The effects of IS on cell viability were determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays and levels of apoptosis-related proteins (Bcl-2-associated protein X [Bax] and B-cell lymphoma 2 [Bcl-2]), nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, and phosphorylation of mitogen-activated protein kinase (MAPK) and protein kinase B (Akt) were determined by semiquantitative immunoblotting. The promoter activity of $NF-{\kappa}B$ was measured by luciferase assays and apoptosis was determined by f low cytometry of cells stained with f luorescein isothiocyanate-conjugated Annexin V protein. Results: IS treatment increased ROS production, decreased cell viability and induced apoptosis in HK-2 cells. IS treatment increased the expression of apoptosis-related protein Bax, decreased Bcl-2 expression, and activated phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt. In contrast, paricalcitol treatment decreased Bax expression, increased Bcl-2 expression, and inhibited phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt in HK-2 cells. $NF-{\kappa}B$ promoter activity was increased following IS, administration and was counteracted by pretreatment with paricalcitol. Additionally, flow cytometry analysis revealed that IS-induced apoptosis was attenuated by paricalcitol treatment, which resulted in decreased numbers of fluorescein isothiocyanate-conjugated Annexin V positive cells. Conclusions: Treatment with paricalcitol inhibited IS-induced apoptosis by regulating MAPK, $NF-{\kappa}B$, and Akt signaling pathway in HK-2 cells.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.117-123
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• 2010

We used a flow cytometer to investigate the variations in endopolyploidy (the frequencies of nuclei with DNA contents equivalent to 4C through 16C) during the short period of the early growing stage in vigorously growing young tissues of maize seedlings. We examined different portions of the root and leaves that had been growing for 7 (day 7) and 13 (day 13) days after germination. Endoreplication showed two opposing phenomena without aging. In one case, the endopolyploidy of the first leaf was higher on day 13 than on day 7. In the latter case, endopolyploidy decreased, as clearly revealed by a comparison of the endopolyploidy of the second leaves and the 160-170 mm portion of the seminal root on days 7 and 13. Endopolyploidy was also lower in the top of the leaf. In roots, endopolyploidy was increased by the exogenous application of abscisic acid for only 1 day. The levels of endopolyploidy increased without an increase in cell size in the roots. These results showed that endoreplication occurs in actively growing and young tissue and that the variation can be induced in the short period examined.