A myofiber of skeletal muscle is composed of myofibrils, sarcolemma (plasma membrane), and constameres, which anchor the myofibrils to the sarcolemma. Achvillin is a recently identified F-actin binding muscle protein, co-isolates with dystrophin and caveolin-3 in low-density sarcolemma of striated muscle, and colocalizes with dystrophin at costameres, the specialized adhesion sites in muscle. Archvillin also binds to nebulin and localizes at myofibrillar Z-discs, the lateral boundaries of the sarcomere in muscle. However other roles of archvillin on the dynamics of myofibrillogenesis remain to be defined. The goal of this study is, by using siRNA-mediated gene silencing technique, to investigate the effect of archvillin on the dynamics of myofibrillogenesis in cell culture of a mouse skeletal myogenic cell line (C2C12), where presumptive myoblasts withdraw from the cell cycle, fuse, undergo de novo myofibrillogenesis, and differentiate into mature myotubes. The roles of archvillin in the assembly and maintenance of myofibril and during the progression of myofibrillogenesis induced in skeletal myoblast following gene silencing in the cell culture were investigated. Fluorescence microscopy demonstrated that the distribution of archvillin was changed along the course of myofibril assembly with nebulin, vinculin and F-actin and then located at Z-lines with nebulin. Fluorescence microscopy demonstrated that knockdown of mouse archvillin expression led to an impaired assembly of new myofibrillar clusters and delayed fusion and myofibrillogenesis although the mouse archvillin siRNA did not affect those expressions of archvillin binding proteins, such as nebulin and F-actin. This result is corresponded with that of RT-PCR and western blots. When the perturbed archvillin was rescued by co-transfection with GFP or Red tagged human archvillin construct, the inhibited cell fusion and myotube formation was recovered. By using siRNA technique, archvillin was found to be involved in early stage of myofibrillogenesis. Therefore, the current data suggest the idea that archvillin plays critical roles on cell fusion and dynamic myofibril assembly.
To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.
The bacterial toxin, tolaasin, causes brown blotch disease on the cultivated mushrooms by collapsing fungal and fruiting body structure of mushroom. Cytotoxicity of tolaasin was evaluated by measuring hemolytic activity because tolaasins form membrane pores on the red blood cells and destroy cell structure. While we investigated the inhibitions of hemolytic activity of tolaasin by $Zn^{2+}$ and $Cd^{2+}$, we found that $Ni^{2+}$ is another antagonist to block the toxicity of tolaasin. $Ni^{2+}$ inhibited the tolaasin-induced hemolysis in a dose-dependent manner and its Ki value was $\sim10$ mM, implying that the inhibitory effect of $Ni^{2+}$ is stronger than that of $Cd^{2+}$. The hemolytic activity was completely inhibited by $Ni^{2+}$ at the concentration higher than 50 mM. The effect of $Ni^{2+}$ was reversible since it was removed by the addition of EDTA. When the tolaasin-induced hemolysis was suppressed by the addition of 20 mM $Ni^{2+}$, the subsequent addition of EDIA immediately initiated the hemolysis. Although the mechanism of $Ni^{2+}$ -induced inhibition on tolaasin toxicity is not known, $Ni^{2+}$ could inhibit any of fallowing processes of tolaasin action, membrane binding, molecular multimerization, pore formation, and massive ion transport through the membrane pore. Our results indicate that $Ni^{2+}$ inhibits the pore activity of tolaasin, the last step of the toxic process.
Objectives : This study was conducted to evaluate the efficacy of a complex mixture of natural substances of ginseng and baeknyeoncho on the arthritic rats. Methods : In vitro experiments were conducted to ensure the stability of the complex. After setting toxicity and concentration by MTT assay, the antioxidant effect was measured through DPPH and ABTS radical scavenging activity. To confirm the anti-inflammatory effects of the complex, levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured in LPS-treated macrophage cell lines (RAW264.7). We injected monosodium iodoacetate (MIA) 50 μl (60 mg/ml) into knee joints of rats to induce osteoarthritis. The rats were divided into three groups (normal (n=5), control (n=5), and OR (n=5) group). The control group consumed 2 mg/kg of physiological saline once a day for 4 weeks, and the OR group was mixed at a concentration of 416.5 mg/kg of Baengnyeoncho (O) and 208.25 mg/kg of red ginseng (R) and ingested 1 mL each 5 days a week. Results : This complex increased the DPPH and ABTS radical scavenging rate. The complex decreased NO production and pro-inflammatory cytokine production of macrophages. In the OR group, the secretion of cytokine in serum was decreased. In histopathological examination, the joint tissue of the composite showed less damage to the synovial membrane, cartilage, and fibrous tissue than the control group. Conclusions : As a result of this study, natural complexes have antioxidant, anti-inflammatory and cartilage protection effects. Therefore, we expect the complex to be effective in treating osteoarthritis.
The RLG (relative length of gut) is 1.52 (n=12) in the sablefish, Anoplopoma fimbria. The digestive tract has five or six pyloric caeca in the posterior region of stomach. Morphology of mucosal fold is unbranched type in the esophagus and stomach, but branched type in the intestine. The histological structure of digestive tract can be divided into mucosal layer, submucosal layer, muscular layer and serous membrane in the cross section. In the esophagus, mucosal epithelial layer is a simple, and consists of ciliated columnar epithelia and mucous cells. In the stomach, gastric gland of mucosal epithelial layer is a tubular, and is composed of chief cell, parietal cell and mucin secreting cell, which is columnar and contained secretory granules of red and blue colors in the AB-PAS (pH 2.5) reaction. In the intestine, mucosal epithelial layer is a simple, and consists of ciliated columnar epithelia and goblet cells. The submucosal layer is composed mainly of collagen fibers, and well developed in the esophagus. And the muscular layer of digestive tract is divided into longitudinal and circular muscle layer, and well developed in the stomach. The liver is composed of numerous lobular structure and bile canaliculi. Stainability of hepatocyte cytoplasm was eosinophilic, and nucleus and nucleolus showed basophilic in the H-E stain. The pancreatic tissue was scattered in the fatty tissue near the digestive tract, and acinar gland consisting of numerous exocrine cells. And cytoplasmic stainability of exocrine cell was basophilic, and contained numerous zymogen granules of eosinophilic in the H-E stain.
Yoon, Se Hoon;Kim, Dong Sup;Yu, Seung Taek;Shin, Sae Ron;Choi, Du Young
Clinical and Experimental Pediatrics
/
v.58
no.1
/
pp.15-19
/
2015
Purpose: Soluble transferrin receptor (sTfR) is a truncated extracellular form of the membrane transferrin receptor produced by proteolysis. Concentrations of serum sTfR are related to iron status and erythropoiesis in the body. We investigated whether serum sTfR levels can aid in diagnosis and treatment of iron deficiency anemia (IDA) in children. Methods: Ninety-eight patients with IDA were enrolled and were classified according to age at diagnosis. Group 1 comprised 78 children, aged 6-59 months, and group 2 comprised 20 adolescents, aged 12-16 years. Results: In group 1, patients' serum sTfR levels correlated negatively with mean corpuscular volume; hemoglobin (Hb), ferritin, and serum iron levels; and transferrin saturation and positively with total iron binding capacity (TIBC) and red cell distribution width. In group 2, patients' serum sTfR levels did not correlate with ferritin levels and TIBC, but had a significant relationship with other iron indices. Hb and serum sTfR levels had a significant inverse relationship in both groups; however, in group 1, there was no correlation between Hb and serum ferritin levels. In 30 patients of group 1, serum sTfR levels were significantly decreased with an increase in Hb levels after iron supplementation for 1 month. Conclusion: Serum sTfR levels significantly correlated with other diagnostic iron parameters of IDA and inversely correlated with an increase in Hb levels following iron supplementation. Therefore, serum sTfR levels can be a useful marker for the diagnosis and treatment of IDA in children.
Kim, Dong-Shoo;Kwon, Hyang-Mi;Choi, Jung-Suk;Kang, Sang-Wook;Ji, Geun-Eog;Kang, Young-Hee
Nutrition Research and Practice
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v.1
no.4
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pp.285-290
/
2007
The leukocyte recruitment and transmigration across the endothelial barrier into the vessel wall are crucial steps in atherosclerosis. Leukocyte trafficking on the endothelium is elicited by induction of endothelial adhesion molecules, and its transmigration is mediated by degradation of basement membrane proteins through enzymatic activity of matrix metalloproteinases (MMP). The current study investigated whether resveratrol, a polyphenol present in grapes and red wine, was capable of inhibiting leukocyte adhesion to tumor necrosis factor (TNF)-${\alpha}$-activated endothelium. It was found that resveratrol inhibited the TNF-${\alpha}$-activated endothelial expression of vascular cell adhesion molecule-1 in a dose-dependent manner. In addition, resveratrol hampered THP-1 monocyte adhesion to activated endothelial cells. This study further examined whether resveratrol interfered with transendothelial migration of leukocytes. The MMP-2 gelatinolytic activity of endothelial cells was enhanced by TNF-${\alpha}$, which was attenuated by an addition of ${\geq}25{\mu}M$ resveratrol. In addition, 25 ${\mu}M$ resveratrol mitigated the MMP-9 activity of THP-1 cells, followed by a marked inhibition of transendothelial migration. These results demonstrated that resveratrol suppressed monocyte adhesion and migration induced by TNF-${\alpha}$ through modulating expression of adhesion molecules and gelatinolytic activity of MMP. These findings suggest that dietary resveratrol may be therapeutic agent for inhibiting leukocyte recruitment into the subendothelium during inflammatory atherosclerosis.
Ki, Mi-Ran;Yun, Soon-Kyu;Choi, Kyung-Min;Hwang, Se-Young
Journal of Microbiology and Biotechnology
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v.21
no.5
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pp.483-493
/
2011
An acid phosphatase (HppA) activated by $NH_4Cl$ was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH${\leq}$4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., $K^+$,$ NH_4{^+}$, and/or $Ni^{2+}$). In particular, $Ni^{2+}$ appeared to lower the enzyme's $K_m$ for the substrates, without changing $V_{max}$. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an $HppA^-$ H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by $NH_4Cl$. In contrast to wild type, $HppA^-$ H. pylori cells grew more slowly. Strikingly, they imported $Mg^{2+}$ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope.
Parasitic leeches could directly (through causing poor growth, anemia and wound in the fish) and indirectly (by predisposition of the fish to secondary bacterial and fungal infections) affects their hosts. In the present study, fishes that were attacked by leeches in natural and experimental environment were studied. Pathologic samples were obtained from damages at the site of leech bite, as well as kidney and liver of the fish. Histopathological examination revealed numerous lesions at the site of leech bite including tissue demolition, detachment at the site of leech bite in the epidermis of epithelial tissue in the skin, destructed nucleus in epithelial cells of the skin plus necrosis in the damaged skin and weak inflammatory penetration to acute necrotic damages along with piercing dermis layer. Pathologic lesions in the kidney included some changes such as proliferation by increasing glomerular cells and membrane cells in capillary vein of the kidney, blood cell necrosis in kidney with infiltration of white blood cells mainly mononuclear and less polymorphonuclear which are the symptoms of anemia due to blood feeding and sucking by leeches. There was also a chronic kidney infection probably originated from another part of body such as skin. Moreover, leeches caused hemorrhagic anemia due to blood consumption of the hosts, which led to observation of immature red blood cells. Also results showed that diseases induced by leeched in fish could be acute or chronic, which depends on size of fish, species of leech and severity of infection.
Kim, Hye Ji;Lee, Seung Hyun;Hong, Young Kyu;Kim, Sung Chul
Korean Journal of Soil Science and Fertilizer
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v.51
no.2
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pp.119-127
/
2018
Veterinary antibiotics (VAs) has been used to treat animal disease and to increase body weight. However, released VAs in the soil via spreading of compost can transport to plant and affect its growth. Main purpose of this research was i) to monitor VAs concentration in plant and ii) to evaluate inhibition effect of VAs residuals on the plant growth. Red lettuce (Lactuca sativa) was cultivated for 35 days in the pot soil spiked with 3 different concertation (0.05, 0.5, $5.0mg\;kg^{-1}$) of chlortetracycline (CTC) and sulfamethazine (SMZ). After 35 days of cultivation, concentration of CTC and SMZ in the plant was measured. Residual of CTC and SMZ was only quantified at the range of $0.007-0.008mg\;kg^{-1}$ and $0.006-0.017mg\;kg^{-1}$ in the leaf and root respectively when high concentration ($5.0mg\;kg^{-1}$) of antibiotic was spiked in the soil. Leaf length and root mass was statistically reduced when $0.05mg\;kg^{-1}$ of CTC was spiked in the soil while no statistical difference was observed for SMZ treatment. This result might indicated that high $K_{ow}$ and $K_d$ value are the main parameters for inhibiting plant growth. Antibiotics that has a high $K_{ow}$ causing hydrophobicity and easy to bioaccumulate in the lipid cell membrane. Also, antibiotics that has a high $K_d$ properties can be sorbed in the root causing growth inhibition of the plant. Overall, management of VAs should be conducted to minimize adverse effect of VAs in the ecosystem.
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