Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.
Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.
Kim Yo Han;Sung Kang Keyng;Lee Kwang Ro;Lee Sang Kwan;Cheong Sang Su;Kang Sei Young;Lee So Young
Journal of Physiology & Pathology in Korean Medicine
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v.17
no.3
/
pp.829-836
/
2003
Seogakjihwang-tang (SJT) was widely used to treat patients suffering from cerebral infarction. But scientific investigation has been carried out very little. The aim of the present study is to investigate the effect of SJT on the production of various cytokines in the patients with cerebral infarction (CI). We investigated interleukin (IL)-4, IL-10 and transforming growth factor (TGF)-1 in the sera of 27 patients with cerebral infarction under consciousness disorders and 10 normal controls using an originally devised sensitive sandwich enzyme-linked immunosorbent assay (ELISA). We found that plasma levels of IL-4 were slightly elevated in patients with cerebral infarction, whereas plasma levels of IL-10 (P<0.001) and TGF-1 were reduced. Peripheral blood mononuclear cells (PBMC) obtained from the patient with CI were cultured for 24 h in the presence or absence of lipopolysaccharide (LPS) or phytohaemagglutinin (PHA). The amount of IL-4, IL-10 and TGF-1, in culture supernatant, was significantly increased in the LPS or PHA treated cells compared to unstimulated cells (P<0.05), We also show that increased cytokines IL-4, and IL-10 level was significantly inhibited by SJT in a dose-dependent manner. Maximal inhibition rate of IL-4 and IL-10 production by SJT was 45.63.3% and 614.7% for LPS-stimulated cell and 27.31.2% and 83.62% for PHA-stimulated cells, respectively (P<0.05). On the other hand, SJT significantly increased the LPS or PHA-induced TGF-1 production (P<0.05). These data suggest that SJT has a regulatory effect on the cytokines production, which might explain its beneficial effect in the treatment of CI.
Shrimp Jeot-Gal is a popular traditional Korean fermented seafood and has been used for seasoning. We isolated a bacterium showing strong extra-cellular fibrinolysis and chitinase activity from shrimp Jeot-Gal and the strain was designated SC082. SC082 was identified as Bacillus licheniformis by 16S rRNA sequence homology search. B. licheniformis SC082 exhibited optimum temperature, pH, and salt concentration at $37^{\circ}C$, pH 7.0, and 6%, respectively. Substrate specificity of the culture supernatant from B. licheniformis SC082 was detected in fibrin, skim milk, and chitin plate. The fibrinolytic activity was highly maintained up to $50^{\circ}C$ at a pH of 7.0 for 3 hr and was stable up to pH 9.0 at $37^{\circ}C$ for 3 hr. The chitinase activity was remarkably induced by addition of 1.0% colloidal chitin and the pH and temperature optima of the enzyme were 5.0 and $45^{\circ}C$, respectively. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram analysis, this strain produced three fibrinolytic isozymes and two chitinase isozymes. The approximate molecular weights of the putative fibrinolytic enzymes were 23.0, 62.0, and 72.0 kDa and those of the chitinases were 62.0 and 55.0 kDa, respectively. The antioxidant activity of SC082 was also measured by using 2,2-diphenyl-l-picryl-hydrazyl (DPPH) free radical. The DPPH radical scavenging was slightly increased in a dose-dependent manner.
Start-ups often use crowdfunding platforms such as Wadiz to get financing from corporate investors and general public. These platforms include functions to increase participant activity through the number of comments, interest and online word-of-mouth. This study aims to elucidate how these functions related to a participant activity exhibit mediating effects on a success of crowdfunding and an achievement rate of targeted investment amount. To this end, the individual variables such as progress period, target amount, venture company, experience of attracting investment, possession of intellectual property rights, a career of the representative and an award experience of the representative were classified into the project characteristics, the company characteristics and the representative characteristics through not only previous studies but also interviews with investment professionals and platform operators. Afterwards, this study went through an empirical verification process using a structural equation model that has both crowdfunding performance and participant activity, which is the independent and mediating factors of three perspectives, as well as the dependent variables. In other words, this study analyzes how the characteristics of three perspectives affect the participant activity and how the participant activity had the mediating effects on the crowdfunding performance. In addition, it derives how the analysis results mentioned above vary according to business types (internet services, culture/art, manufacturing/distribution) and investment styles (stock type and bond type). It is expected that this study will help not only in deriving the factors affecting the performance of an investment type crowdfunding but also in preparing the measures to increase a participant activity.
The flowering in Lemna gibba G3, a long-day plant, was promoted under continuous light by agmatine, putrescine, spermidine and spermine present in the culture medium. Methylglyoxal-bis (guanylhydrazone) (MGBG) and cyc10hexylamine (CHA), inhibitors of polyamine biosynthesis, were found to suppress the flowering in the plants. The vegetative grov.1h rate was kept constant while the flowering was being promoted by the pOlyamines, and the inhibitors with depressive effect on flowering showed stimulatory effect on vegetative grov.1h. The pattern of vegetative growth during floral promotion or depression was an indication that the promotive action of the pOlyamines and the suppressive effect of the inhibitors may be outcome of their possible involvement specifically in the flowering process rather than in broad spectrum of growth of L. gibba G3. The degree of promotive action of spermdine and spermine could not be altered (or lessened) by simultaneous application of their inhibitors to the medium. This phenomenon indicates that the flowering process in L. gibba G3 may largely be dependent to the status of endogenous spermidine and spermine. Endogenous level of spermidine in florally induced Lemna, was found to rapidly increase. In 24 h of floral induction, the content reached at the level 2 times higher than that in non-induced plants. The elevated level of spermidine provides an additional, though premature, evidence supporting the postulation that endogenous polyamine status might play an important role in the very early stage of floral induction in L. gibba G3.bba G3.
Among poly aromatic hydrocarbons, dioxin and PCBs are the most controversial environmental pollutants in our modern life. These pollutants are known as human carcinogens, and liver is the most sensitive target in animal cancer models. Specific aims of the study were focused on the mechanism of carcinogenesis in hepatocytes and the structure-activity relation among these diverse environmental chemicals. Because key mechanisms of dioxin-induced carcinogenesis in human epithelial cell model are the alteration of signal transduction pathway and PKC isoforms, the alteration of the signal transduction pathways and other factors associated with carcinogenesis were studied. Rat hepatocytes cultured under the sandwich protocols were exposed with the various concentration of dioxins and PCBs, and signal transduction pathway, protein kinase C isoforms, oxidant stress, and apoptotic nuclei were evaluated. Since it is important to understand the structure-activity relation among these chemicals to properly assess the carcinogenic potentials, the study analyzed the parameters associated with carcinogenic processes, based on their structural characteristics. In addition, signal transduction pathways and PKC isoforms involved in inhibition of UV-induced apoptosis were also analyzed to elaborate the tumor promotion mechanism of these chemicals. Induction of apoptosis by UV irradiation was optimal at $60\;J/m^2$ in primary hepatocyte in culture. Compared to non coplanar PCBs such as PCB 114 and PCB 153, coplanar PCBs such as PCB 77 and PCB126 showed a stronger inhibition of apoptosis induced by UV irradiation. Production of reactive oxygen species (ROS) was more stimulated by non-coplanar PCBs than coplanar PCBs with the most potent induction of ROS by chlorinated non-coplanar PCB. As compared to the level of induction by PCB126, non-coplanar PCB153 showed a higher increase of intracellular concentrations. Besides the alteration of intracellular calcium concentration, translocation of PKC from cytosolic fraction to membrane fraction was clearly observed upon the exposure of non-coplanar PCB. Taken together, the present study demonstrated that there is a potent structure-activity relationship among PCB congeners and the mechanism of PAH-induced carcinogenesis is structure-specific. The study suggested that more diverse pathways of PAH-induced carcinogenesis should be taken into account beyond the boundary of Ah receptor dogma to assess the health impact of PAH with more accuracy.
The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.
Jung, A Young;Jung, Ki Youn;Lin, Chunmei;Yon, Jung-Min;Lee, Jong Geol;Lee, Beom Jun;Yun, Young Won;Nam, Sang-Yoon
Korean Journal of Veterinary Research
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v.55
no.2
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pp.133-139
/
2015
The increasing uses of zinc oxide nanoparticles (nZnO) in industrial and personal care products raise possible danger of using nZnO in human. To determine whether ZnO induces size-dependent anomalies during embryonic organogenesis, mouse embryos on embryonic day 8.5 were cultured for 2 days under 50, 100, and $150{\mu}g$ of nZnO (< 100 nm) or micro-sized ZnO (mZnO; $80{\pm}25{\mu}m$), after which the morphological changes, cumulative quantity of Zn particles, and expressions of antioxidant and apoptotic genes were investigated. Although embryos exposed to $50{\mu}g$ of ZnO exhibited no defects on organogenesis, embryos exposed to over $100{\mu}g$ of ZnO showed increasing anomalies. Embryos treated with $150{\mu}g$ of nZnO revealed significant changes in Zn absorption level and morphological parameters including yolk sac diameter, head length, flexion, hindbrain, forebrain, branchial bars, maxillary process, mandibular process, forelimb, and total score compared to the same dose of mZnO-treated embryos. Furthermore, CuZn-superoxide dismutase, cytoplasmic glutathione peroxidase (GPx) and phospholipid hydroperoxidase GPx mRNA levels were significantly decreased, but caspase-3 mRNA level was greatly increased in nZnO-treated embryos as compared to normal control embryos. These findings indicate that nZnO has severer teratogenic effects than mZnO in developing embryos.
Park, Ji-Young;Lee, Tae-Ho;Oh, You-Kwan;Kim, Jun-Rae;Seol, Eun-Hee;Jung, Gyoo-Yeol;Kim, Mi-Sun;Park, Sung-Hoon
KSBB Journal
/
v.20
no.6
/
pp.458-463
/
2005
[ $H_2$ ] from CO and water was continuously produced in a trickle bed reactor(TBR) using Citrobacter amalonaticus Y19. When the strain C. was cultivated in a stirred-tank reactor under a chemoheterotrophic and aerobic condition, the high final cell concentration of 13 g/L was obtained at 10 hr. When the culture was switched to an anaerobic condition with the continuous supply of gaseous CO, CO-dependent hydrogenase was fully induced and its hydrogen production activity approached 16 mmol/g cell/hr in 60 hr. The fully induced C. amalonaticus Y19 cells were circulated through a TBR packed with polyurethane foam, and the TBR was operated for more than 20 days for $H_2$ production. As gas retention time decreased or inlet CO partial pressure increased, $H_2$ production rate increased but the conversion from CO to $H_2$ decreased. The maximum $H_2$ production rate obtained was 16 mmol/L/hr at the gas retention time of 25 min and the CO inlet partial pressure of 0.4 atm. The high $H_2$ production rate was attributed to the high cell density in the liquid phase circulating the TBR as well as the high surface area of polyurethane foam used as packing material of the TBR.
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