• The Journal of Internal Korean Medicine
• /
• v.21 no.5
• /
• pp.705-713
• /
• 2000

Objectives : The purpose of this study was to investigate the effect of Intravascular Laser Irradiation of Blood(ILlB) by the live blood analysis. Methods : We had analysed the changing forms of the live blood samples with Ultra Darkfield Microscope before and after Intravascular Laser Irradiation of Blood. Results : 1. Somatid did not showed significant change. 2. In the rouleau of red blood cells was decreased significantly. 3. In the morphological change of red blood cells, Burr cell, Ovalocyte and Poikilocyte were decreased significantly, but Acanthocyte and Target cell were increased significantly. 4. In the abnormal matters in plasma, the Cholesterol cristal did not showed significant change, but the Aggregation of platelet, Lipids, Spicule, Leucocyte, Uric acid cristal did showed a little significant decrease. Conclusion : These findings suggest that live blood analysis is useful to judge the effect of treatment and diagnosis in oriental medicine, and with the effect of Intravascular Laser Irradiation of Blood, it had showed significant effect on rouleau of red blood cells, morphological change of red blood cells and abnormal matters in plasma.

• BMB Reports
• /
• v.38 no.6
• /
• pp.639-645
• /
• 2005

Protein kinase C (PKC) isozymes, a family of serine-threonine kinases, are important regulators of cell proliferation and malignant transformation. Phorbol esters, the prototype PKC activators, cause PKC translocation to the plasma membrane in prostate cancer cells, and trigger an apoptotic response. Studies in recent years have determined that each member of the PKC family exerts different effects on apoptotic or survival pathways. $PKC{\delta}$, one of the novel PKCs, is a key player of the apoptotic response via the activation of the p38 MAPK pathway. Studies using RNAi revealed that depletion of $PKC{\delta}$ totally abolishes the apoptotic effect of the phorbol ester PMA. Activation of the classical $PKC{\alpha}$ promotes the dephosphorylation and inactivation of the survival kinase Akt. Studies have assigned a pro-survival role to $PKC{\varepsilon}$, but the function of this PKC isozyme remains controversial. Recently, it has been determined that the PKC apoptotic effect in androgen-dependent prostate cancer cells is mediated by the autocrine secretion of death factors. $PKC{\delta}$ stimulates the release of $TNF{\alpha}$ from the plasma membrane, and blockade of $TNF{\alpha}$ secretion or $TNF{\alpha}$ receptors abrogates the apoptotic response of PMA. Molecular analysis indicates the requirement of the extrinsic apoptotic cascade via the activation of death receptors and caspase-8. Dissecting the pathways downstream of PKC isozymes represents a major challenge to understanding the molecular basis of phorbol ester-induced apoptosis.

• YAKHAK HOEJI
• /
• v.54 no.5
• /
• pp.354-361
• /
• 2010

Glycosphingolipids are structural components of mammalian cell membranes and are involved in essential cellular physiology such as cell-cell interaction, recognition, transmembrane signaling, proliferation and cell death. In this study, the simple quantitative method of monoglycoceramides-containing glucosylceramide and galactosylceramide was developed. The glycosylceramides extracted from culture cells and rat plasma were resolved by TLC, deacylated by SCDase and analyzed by HPLC-fluorescence detector at an excitation wavelength of 340 nm and an emission wavelength of 455 nm. Limit of detection was approximately 0.1 pmol and limit of quantification was about 1 pmol for both monoglycoceramide standards. The recoveries of standard glucosylceramides from intra- and inter-day assays were 113.8 and 88.8% and those of galactosylceramides were 110.7 and 123.9%, respectively. The monoglycoceramide contents of SW-620 cells and rat plasma were $141.5{\pm}5$ pmol/$1{\times}10^6$ cells and $3.9{\pm}0.3{\mu}M$, respectively. The present analytical method provides a reproducible quantification and total content of monoglycoceramide which may be as a potential biomarker for lipid imbalance-related human diseases.

• IMMUNE NETWORK
• /
• v.14 no.4
• /
• pp.207-218
• /
• 2014

Chronic virus infection leads to the functional impairment of dendritic cells (DCs) as well as T cells, limiting the clinical usefulness of DC-based therapeutic vaccine against chronic virus infection. Meanwhile, B cells have been known to maintain the ability to differentiate plasma cells producing antibodies even during chronic virus infection. Previously, ${\alpha}$-galactosylceramide (${\alpha}GC$) and cognate peptide-loaded B cells were comparable to DCs in priming peptide-specific $CD8^+$ T cells as antigen presenting cells (APCs). Here, we investigated whether B cells activated by ${\alpha}GC$ can improve virus-specific T cell immune responses instead of DCs during chronic virus infection. We found that comparable to B cells isolated from naïve mice, chronic B cells isolated from chronically infected mice with lymphocytic choriomeningitis virus (LCMV) clone 13 (CL13) after ${\alpha}GC$-loading could activate CD1d-restricted invariant natural killer T (iNKT) cells to produce effector cytokines and upregulate co-stimulatory molecules in both naïve and chronically infected mice. Similar to naïve B cells, chronic B cells efficiently primed LCMV glycoprotein (GP) 33-41-specific P14 $CD8^+$ T cells in vivo, thereby allowing the proliferation of functional $CD8^+$ T cells. Importantly, when ${\alpha}GC$ and cognate epitope-loaded chronic B cells were transferred into chronically infected mice, the mice showed a significant increase in the population of epitope-specific $CD8^+$ T cells and the accelerated control of viremia. Therefore, our studies demonstrate that reciprocal activation between ${\alpha}GC$-loaded chronic B cells and iNKT cells can strengthen virus-specific T cell immune responses, providing an effective regimen of autologous B cell-based therapeutic vaccine to treat chronic virus infection.

• The Korean Journal of Cytopathology
• /
• v.12 no.1
• /
• pp.67-71
• /
• 2001

Primary malignant lymphoma of the thyroid gland is uncommon malignancies. Its fine needle aspiration cytology (FNAC) findings are rarely described in the literature. This article highlights the FNAC diagnosis of primary malignant lymphoma of the thyroid gland. A 70-year-old female presented with a rapidly enlarging thyroid mass of five months' duration. FNAC smears showed low cellularity consisting of predominantly atypical enlarged lymphoid cells admixed with a few small lymphocytes, plasma cells, and oncocytic cells. Some disrupted lymphoid cells were also present. The tumor cells infiltrated into the thyroid follicular epithelium forming lymphoepithelial lesion, The cytologic appearance showed a diffuse mixture of cell types with only a few small, mature lymphocytes and many enlaraed lymphoid cells. The enlarged lymphoid cells were atypical and pleomorphic with nuclear clefting and irregularities. Grossly, the left lobe of the thyroid was nearly replaced by a diffuse firm to soft solid mass with smooth tan fish-flesh homogeneous cut surface. Histological diagnosis was diffuse large B-cell lymphoma with areas of marginal zone B-cell lymphoma of MALT type.

• The Korean Journal of Physiology and Pharmacology
• /
• v.16 no.1
• /
• pp.71-77
• /
• 2012

Mitochondrial dynamics and distribution is critical for their role in bioenergetics and cell survival. We investigated the consequence of altered fission/fusion on mitochondrial function and motility in INS-1E rat clonal ${\beta}$-cells. Adenoviruses were used to induce doxycycline-dependent expression of wild type (WT-Mfn1) or a dominant negative mitofusin 1 mutant (DN-Mfn1). Mitochondrial morphology and motility were analyzed by monitoring mitochondrially-targeted red fluorescent protein. Adenovirus-driven overexpression of WT-Mfn1 elicited severe aggregation of mitochondria, preventing them from reaching peripheral near plasma membrane areas of the cell. Overexpression of DN-Mfn1 resulted in fragmented mitochondria with widespread cytosolic distribution. WT-Mfn1 overexpression impaired mitochondrial function as glucose- and oligomycin-induced mitochondrial hyperpolarization were markedly reduced. Viability of the INS-1E cells, however, was not affected. Mitochondrial motility was significantly reduced in WT-Mfn1 overexpressing cells. Conversely, fragmented mitochondria in DN-Mfn1 overexpressing cells showed more vigorous movement than mitochondria in control cells. Movement of these mitochondria was also less microtubule-dependent. These results suggest that Mfn1-induced hyperfusion leads to mitochondrial dysfunction and hypomotility, which may explain impaired metabolism-secretion coupling in insulin-releasing cells overexpressing Mfn1.

• Journal of Microbiology
• /
• v.43 no.3
• /
• pp.266-271
• /
• 2005

In eukaryotic cells, various proteins are anchored to the plasma membrane through glycosylphosphatidylinositol (GPI). To study the biosynthetic pathways and modifications of GPI, various mutant cells have been isolated from the cells of Chinese hamster ovaries (CHO) supplemented with several exogenous genes involved in GPI biosynthesis using aerolysin, a toxin secreted from gram-negative bacterium Aeromonas hydrophila. Alpha toxin from Gram-positive bacterium Clostridium septicum is homologous to large lobes (LL) of aerolysin, binds GPI-anchored proteins and possesses a cell-destroying mechanism similar to aerolysin. Here, to determine whether alpha toxins can be used as an isolation tool of GPI-mutants, like aerolysin, CHO cells stably transfected with several exogenous genes involved in GPI biosynthesis were chemically mutagenized and cultured in a medium containing alpha toxins. We isolated six mutants highly resistant to alpha toxins and deficient in GPI biosynthesis. By genetic complementation, we determined that one mutant cell was defective of the second subunit of dolichol phosphate mannose synthase (DPM2) and other five cells were of a putative catalytic subunit of inositol acyltransferase (PIG-W). Therefore, C. septicum alpha toxins are a useful screening probe for the isolation of various GPI-mutant cells.

• IMMUNE NETWORK
• /
• v.21 no.1
• /
• pp.10.1-10.13
• /
• 2021

The coronavirus disease 2019 (COVID-19) pandemic (severe acute respiratory syndrome coronavirus 2) is a global infectious disease with rapid spread. Some patients have severe symptoms and clinical signs caused by an excessive inflammatory response, which increases the risk of mortality. In this study, we reanalyzed scRNA-seq data of cells from bronchoalveolar lavage fluids of patients with COVID-19 with mild and severe symptoms, focusing on Ab-producing cells. In patients with severe disease, B cells seemed to be more activated and expressed more immunoglobulin genes compared with cells from patients with mild disease, and macrophages expressed higher levels of the TNF superfamily member B-cell activating factor but not of APRIL (a proliferation-inducing ligand). In addition, macrophages from patients with severe disease had increased pro-inflammatory features and pathways associated with Fc receptor-mediated signaling, compared with patients with mild disease. CCR2-positive plasma cells accumulated in patients with severe disease, probably because of increased CCL2 expression on macrophages from patients with severe disease. Together, these results support the hypothesis that different characteristics of B cells might be associated with the severity of COVID-19 infection.

• Journal of Periodontal and Implant Science
• /
• v.25 no.1
• /
• pp.167-178
• /
• 1995

It has been believed that the increased release of free oxygen radicals and their tissue damaging potency might be a contributing factor in the pathogenesis of periodontal disease. Antioxidant enzymes such as superoxide dismutase(SOD) and catalase can protect the tissue damage from the free oxygen radicals($O_2^-,H_2O_2$, and $OH^-$). In order to investigate the SOD- and catalase - activity in the blood plasma and red blood cells(RBCs) of the patients with perodontitis, 19 male periodontitis patients($25{\sim}35$ years old) who had good general health, more than 10 teeth with severely inflamed gingiva, attachment loss more than 6mm and bone loss were selected as periodontitis group, and 13 male volunteers($22{\sim}29$ years old) with good general and periodontal health were selected as normal group. After blood plasma and RBC were separated from peripheral blood of 2ml collected from antecubital vein of each subject, SOD- activity in blood plasma and RBCs was measured by the same method that Paoletti et al. did, and catalase - activity in RBC was measured by the same method that Beers et al, did. The difference of SOD- and catalase - activity between the normal and the periodontitis groups was statistically analyzed by Student t-test with SPSS/PC program.The results were as follows : 1. SOD activity in blood plasma was significantly lower in the periodontitis group($1.986{\pm}0.893$) than in the normal group($3.324{\pm}1.044$)(p<0.05). 2. There was no statistical significance in the difference of SOD- activity in RBCs between the periodontitis group($7.753{\pm}3.206$) and the normal group($8.116{\pm}1.192$)(p$242.8{\pm}45.6$) than in the normal group($280.2{\pm}32.6$)(p

• KSBB Journal
• /
• v.27 no.5
• /
• pp.308-312
• /
• 2012

Argon-plasma jet (Ar-PJ) is generated by ionizing Ar gas, and the resulting Ar-PJ consists of a mixture of neutral particles, positive ions, negative electrons, and various reactive species. Although Ar-PJ has been used in various biomedical applications, little is known about the biological effects on cells located near the plasma-exposed region. Here, we investigated the effects of the Ar-PJ on actin cytoskeleton of mouse embryonic fibroblasts (MEFs) in response to indirect as well as direct exposure to Ar-PJ. This Ar-PJ was generated at 500 mL/min of flow rate and 100 V electric power by our device mainly consisting of electrodes, dielectrics, and a high-voltage power supply. Because actin cytoskeleton is the key cellular machinery involved in cellular movement and is implicated in regulation of cancer metastasis and thus resulting in a highly desirable cancer therapeutic target, we examined the actin filament architectures in Ar-PJ-treated MEFs by staining with an actin-specific phalloidin labeled with fluorescent dye. Interestingly, the Ar-PJ treatment causes destabilization of actin filament architectures in the regions indirectly exposed to Ar-PJ, but no differences in MEFs treated with Ar gas alone and in untreated cell control, indicating that this phenomenon is a specific cellular response against Ar-PJ in the live cells, which are indirectly exposed to Ar-PJ. Collectively, our study raises the possibility that Ar-PJ may have potential as anti-cancer drug effect through direct destabilization of the actin cytoskeleton.