• IMMUNE NETWORK
• /
• 제2권2호
• /
• pp.79-85
• /
• 2002

Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

• BMB Reports
• /
• 제30권4호
• /
• pp.299-301
• /
• 1997

The asialoglycoprotein receptor (ASGPR) was the first described mammalian lectin that mediates the specific binding and internalization of galactose/N-acetylgalactosamine-terminating glycoproteins by hepatic parenchymal cells. H1 and H2 are known as essential subunits of the functional ASGPR. There were close similarities in ASGPR H2 subunits between cultured cell line HepG2 and normal human liver cells including identical sequences at both termini. It was therefore expected that there may be some similarities between the subunits from normal liver cells and fetal liver cells. The two subunits of human fetal liver ASGPR. designated FL-H1 and FL-H2. were cloned from cDNA library by peR and the sequences were compared with the known HI and H2 sequences of HepG2, and the H1 sequence of nornal human liver cells. The results showed that FL-H1 was identical to H1 of HepG2. Whereas FL-H2 contains a 15-bp miniexon, but missing 57-bp at the near upstream from the membrane-spanning domain compared to H2 of HepG2 and normal human liver cells indicating that FL-H2 resulted from a differential splicing compared to HepG2 and normal liver cells.

• Biomolecules & Therapeutics
• /
• 제22권2호
• /
• pp.93-99
• /
• 2014

The interaction between viral HA (hemagglutinin) and oligosaccharide of the host plays an important role in the infection and transmission of avian and human flu viruses. Until now, this interaction has been classified by sialyl(${\alpha}2-3$) or sialyl(${\alpha}2-6$) linkage specificity of oligosaccharide moieties for avian or human virus, respectively. In the case of H5N1 and newly mutated flu viruses, classification based on the linkage type does not correlate with human infection and human-to-human transmission of these viruses. It is newly suggested that flu infection and transmission to humans require high affinity binding to the extended conformation with long length sialyl(${\alpha}2-6$)galactose containing oligosaccharides. On the other hand, the avian flu virus requires folded conformation with sialyl(${\alpha}2-3$) or short length sialyl(${\alpha}2-6$) containing trisaccharides. This suggests a potential future direction for the development of new species-specific antiviral drugs to prevent and treat pandemic flu.

• 대한화장품학회:학술대회논문집
• /
• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
• /
• pp.85-96
• /
• 2003

Normal human skin, constantly challenged by environmental micro-organisms, has an innate ability to fight invading microbes through antimicrobial peptides. These peptides, described in both plant and animal kingdoms are able to inactivate a broad spectrum of micro-organisms. Mammalian defensins constitute one of the most common antimicrobial peptide family. Among the three human beta-defensins hBD1, hBD2 and hBD3 produced in epithelia, only hBD2 and hBD3 are inducible and additionally have been described as expressed by differentiated keratinocytes at site of inflammation and infection. The aims of these studies were to define a cell culture model in which the basal production of hBD could be detected and up-regulated in order to enhance skin auto-protection against micro-organisms. A specific Polymerase Chain Reaction method have been developed for hBD2 and hBD3 mRNA detection in non-differentiated monolayer keratinocytes cell culture. We have been able to demonstrate that in vitro, hBD2 and hBD3 expression in normal human keratinocytes could be detected and enhanced by TNF-alpha and IFN-gamma, in hypercalcic culture conditions. This research opened the possibility of the development of cosmetic active compounds, able to induce the expression of skin natural antibiotic peptides responsible about microflora ecology of the skin.

• 대한의생명과학회지
• /
• 제14권3호
• /
• pp.193-198
• /
• 2008

In order to evaluate on the effect of kaempferol on the cytotoxicity of oxygen tree radicals, XTT assay was performed to determine the cell viability after skin fibroblasts derived from human (Detroit 51) that were treated with various concentrations of hydrogen peroxide $(H_2O_2)$. And also, the effect of kaempferol on the cytotoxicity induced by H202 that was examined by cell viability, lactate dehydrogenase (LDH) activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. $H_2O_2$ decreased cell viability in dose-dependent manner in these cultures and the $XTT_{90}\;and\;XTT_{50}$ values were determined at concentration of $35{\mu}M\;and\;90{\mu}M$ of $H_2O_2$ after skin fibroblasts derived from human were treated with $15{\sim}90{\mu}M$ of $H_2O_2$ for 6 hours, respectively. $H_2O_2$ was highly toxic on cultured skin fibroblasts derived from human by toxic criteria of Brenfreund and Puerner (1984). In the protective effect of kaempferol on $H_2O_2$-induced cytotoxicity, kaempferol increased DPPH radical scavenging activity and significantly decreased LDH activity. From these results, it is suggested that oxygen tree radical, $H_2O_2$, was highly toxic on cultured skin fibroblasts derived from human, and also kaempferol of flavonoid showed the protection on $H_2O_2$-induced cytotoxicity.

• Clinical and Experimental Reproductive Medicine
• /
• 제27권4호
• /
• pp.359-366
• /
• 2000

The present studt was performed to investigate the effect of treatment and samples of human follicular fluid (hFF) on the development in vitro of mouse embryos. The two cell stage embryos collected at 40 h post-hCG injection were cultured in the modified human tubal fluid (m-RTF) containing 15% synthetic serum substitute (SSS) or human tubal fluid (hFF) for up to 3 days at $37^{\circ}C$ in 5% $CO_2$ incubator. Also the composition of hormone, total protein and protein pattern of hFF samples were analyzed. The developmental rate of mouse embryos developed to blastocyst were not significant difference in the m-RTF containing 15% hFF filtered with 0.22 or 0.8 ${\mu}m$ syringe filter, however, the embryos cultured in the m-RTF containing inactivated hFF were significantly (p<0.05) developed at the high rate to blastocyst than those containing fresh hFF and SSS. The in vitro developmental rate to blastocyst and hatched blastocyst in the m-RTF containing 15% hFF sample A (90.5 and 85.4%, respectively) and SSS (79.4 and 75.3, respectively) were significantly (p<0.05) increased, compared with hFF sample B (64.2 and 54.1 %, respectively). The hFF sample A tended to be higher concentration of LH, FSR, total protein and the ratio of progesterone/$E_2$ and lower concentration of $E_2$ and progesterone than the hFF sample B, but there were no differences in the protein pattern between the two hFF samples. The results of these study suggest that the addition of hFF to the culture medium enhances the development in vitro to blastocyst and hatched blastocyst, but the in vitro developmental rate of mouse embryos is different between hFF samples.

• Journal of Pharmaceutical Investigation
• /
• 제4권1_2호
• /
• pp.25-30
• /
• 1974

The adsorption of nalidixic acid on human erythrocytes was found to expressed by Freundlich's isotherm. The amount of adsorption of nalidixic acid on erythrocytes increased with an increase of pH. The adsorption of nalidixic acid on human plasma was found to expressed at Scatchard's equation by the equilibrium dialysis method. An influence of pH on adsorption of nalidixic acid to human plasma proteins was studies at pH 4-10. It was found that the degree of adsorption increase with the increase of pH from 4-6, but descreased above pH 9.

• 한국산업보건학회지
• /
• 제19권3호
• /
• pp.261-269
• /
• 2009

To evaluate the effect of antioxidant on the cytotoxicity induced by oxidative stress of reactive oxygen species (ROS) in cultured human skin melanocytes, colorimeric assay of XTT and tyrosinase activity assay were adopted after human skin melanocytes were preincubated for 2 hours in the media containing various concentrations of superoxide dismutase (SOD) before the treatment of hydrogen peroxide. Light microscopic study was carried out in same cultures. The results of this study were as follows 1. Cell viability of human skin melanocytes was significantly decreased by 30 and $40{\mu}M$ of hydrogen peroxide($H_2O_2$), respectively. 2. XTT50 was determined at $30{\mu}M$ after human skin melanocytes were treated with $10{\sim}40{\mu}M$ of hydrogen peroxide for 6 hours. 3. The cell viability of cultured human skin melanocytes pretreated with SOD was increased than that of cultured human skin melanocytes treated with $H_2O_2$ dose-dependently. 4. In tyrosinase activity of human skin melanocytes, the cell treated with SOD showed brown stain compared with $H_2O_2$ treated cells, dark stain. 5. In light microscopy, cultured human skin melanocytes exposed to $H_2O_2$ showed morphological changes such as the decreased cell number and cytoplasmic processes, compared with control. 6. In light microscopy, cultured human skin melanocytes pretreated with SOD showed the increase of cell number and cytoplasmic processes compared with $H_2O_2-treated$ group. From these results, it is suggested that oxidative stress of ROS such as $H_2O_2$ has cytotoxicity by showing the decreased cell viability, the increased tyrosinase activity and mophological changes of the decreased cell number and cytoplasmic processes. While, antioxidant like SOD was effective in the prevention of oxidative stress-mediated cytotoxicity by the increased cell viability, decreased tyrosinase activity and the protection of degenerative morphological changes in cultured human skin melanocytes.

• 대한의생명과학회지
• /
• 제14권1호
• /
• pp.33-38
• /
• 2008

In order to examine oxidative stress of reactive oxygen species and the antioxidant effect of Citri Reticulatae Pericarpium (CRP) extract, human skin melanoma cells were treated with various concentrations of hydrogen peroxide ($H_2O_2$). Antioxidant effect of CRP extract on $H_2O_2$-induced cytotoxicity, cell viability, DPPH-radical scavenging activity and superoxide dismutase (SOD)-like activity. In this study, $H_2O_2$ decreased cell viability of cultured human skin melanoma cells in dose- and time-dependent manners, and then, midcytotoxicity value (MCV) was determined at $60\;{\mu}M$ after human skin melanoma cells were cultured for 5 hours in the media containing $20{\sim}60\;{\mu}M$ of $H_2O_2$, respectively. The $H_2O_2$ was on cultured human skin melanoma cells because MCV of $H_2O_2$ was lower than $100\;{\mu}M$. In the antioxidant effect of CRP extract, CRP extract increased cell viability DPPH-radical scavenging activity and SOD-like activity. From these results, it is suggested that $H_2O_2$ was very toxic on cultured human skin melanoma cells. And also, CRP extract has the antioxidant effect on $H_2O_2$-induced cytotoxicity.

• Toxicological Research
• /
• 제18권4호
• /
• pp.411-416
• /
• 2002

Apoptosis, or programmed cell death, is a genetically regulated pathway that is altered in many cancers. Overexpression of bcl-2 leads to resistance to apoptosis and promotes tumorigenesis. To determine the effect of bcl-2 antisense treatment in human lung cancer cell lines, a 20 mer full phosphorothioate oligonucleotide (ODN) targeted at the coding region of the bcl-2 mRNA was synthesized. Western blot analyses were used to examine bcl-2 protein level in five human non-small cell lung cancer (NSCLC) cell lines (NCI-H226, SK-MES-1 NCI-H358, NCI-H522 and NCI-Hl 299) and four human small cell lung cancer (SCLC) cell lines (NCI-H69, NCI-H4l7, HCC-2108 and SW2). Three out of five NSCLC (NCI-H226, SK-MES-1 and NCI-Hl 299) and all of SCLC cell lines expressed Bcl-2 protein. Treatment of these cell with antisense ODN for 48 hours reduced their viability and Bcl-2 protein level. As a conclusion, bcl-2 antisense treatment appears reduction of the Bcl-2 protein levels and cytotoxic effect including apoptosis in human lung cancer cell lines.