• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.577-583
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• 2007

It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

• Journal of the Korea Society of Computer and Information
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• v.25 no.10
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• pp.115-123
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• 2020

In this paper, we propose to evaluate the potential anti-inflammatory properties of root bark extract of Cudrania (C.) tricuspidata on lipopolysaccharide (LPS)-induced inflammation in macrophages of apolipoprotein E (ApoE) knockout (ApoE^-/-) mice, murine model of atherosclerosis. Atherosclerosis is a chronic vascular inflammatory disease. C. tricuspidata is a small tree of the Moraceae family and its extract has anti-inflammatory activities. However, its role in the progress of atherosclerosis is not yet clear. To determine anti-inflammatory effects of C. tricuspidata in atherogenesis, we applied LPS in peritoneal macrophages of ApoE^-/- mice and measured cell viability by CCK-8 and expression of pro-inflammatory cytokines by qRT-PCR following treatment with root bark extract of C. tricuspidata. Research data was expressed as differences between the cells treated with LPS and root bark extract and the cells treated with LPS alone (control) by a two-tailed non-parametric Mann-Whitney U-test using GraphPad Instat program. No cytotoxic effect was observed when the cells were treated with the extract at concentrations ≤ 100 ㎍/mL. The expression of inflammatory cytokines, including MCP-1, IL-1β, IFN-γ, TNF-α, and IL-6 were inhibited by the extract. These results indicated that the extract has an anti-inflammatory effect and therefore a possible role in the treatment of atherosclerosis.

• Journal of Haehwa Medicine
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• v.24 no.2
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• pp.35-57
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• 2016

Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, $100{\mu}g/ml$). 4. NO, IL-6, TNF-${\alpha}$, MCP-1 production were significantly decreased in GK extract(at 10, $100{\mu}g/ml$). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, $100{\mu}g/ml$). IL-$1{\beta}$, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.

• Journal of Food Hygiene and Safety
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• v.22 no.3
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• pp.145-150
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• 2007

Conjugated linoleic acid (CLA) reduces fat deposition in several mammalian species. The proposed mechanisms for this effect are reduced preadipocyte proliferation and differentiation. The objective of this study was to investigate the inhibitory effects of diglyceride (DG), CLA, DG-CLA of proliferation and differentiation of 3T3-L1 preadipocytes. Cell viability was determined using WST-8 analysis and cell differentiation was determined by glycerol-3-phosphate dehydrogenase (GPDH) activity. Lipid accumulation in differentiating 3T3-L1 cells was measured by Oil red O staining. The proliferation of preconfluent 3T3-L1 cells by treatments of DG, CLA, and DG-CLA was reduced in a dose-dependent manner. CLA among them was the most effective in reduction of viable cells with increasing concentrations. Treatments of the DG, CLA, and DG-CLA at the concentration of $100{\cdot}\ddot{I}g/ml$ for 48h significantly inhibited differentiation of 3T3-L1 cells (p<0.05). In addition. cytoplasmic lipid accumulation during differentiation of the 3T3-L1 preadipocytes was also inhibited by treatments of the test solutions. DG-CLA was the most effective in the inhibition of differentiation and lipid accumulation in 3T3-L1 cells. These results indicate that the DG including CLA as fatty acids is more effective for anti-obesity than DG or CLA alone and that consumption of DG-CLA as a dietary oil may give a benefit for controlling overweight in humans.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.3
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• pp.221-228
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• 2001

This study was carried out to investigate the effect of germination condition and drying temperature on growth and physicochemical properties of brown rice. Three brown rice seeds of Ilpumbyeo, Dasanbyeo and Heugjinjubyeo were stored at room temperature for six weeks to test the time-sequence germination viability. Relatively stable germination ratio was maintained until 2 weeks after storage. However, 3 weeks after storage, germination ratio of brown rice seeds started to decrease rapidly and their germination ratio was lower than 80%. For this reason, brown rice was recommended for seeding within 2 weeks after hulling. During the initial 5 days, germination ratio of 24 hours pre-soaking brown rice was higher about 2-3% than that of non-soaking brown rice. The $25^{\circ}C$ was considered as the most favorable temperature for brown rice germination, because of the high germination ratio and desirable coleoptile growth of the brown rice, and little seed rotting symptoms. The scanning electron micrographs showed the structural differences between hot-air dried and freeze dried germinated-brown rice kernel. In the freeze dried germinated-brown rice, seed coat (pericarp, tegmen and aleurone layer) was mechanically disrupted from the endosperm, and many cleavages were observed among starch storing cells and starch granules. The endosperm of freeze-dried brown rice kernels formed the sponge-like structures and showed the fragile traits. For this reason, hot-air drying is considered as more suitable method than freeze drying for germinated-brown rice. The crude protein and amylose contents were slightly changed, but there were no significant differences during the germination period. Crude fiber content was decreased, but crude Int and total amino acid contents were increased as seeding days increased. A rapid increase in $\alpha$-amylase activities of germinating brown rice was observed at S days after seeding, and $\alpha$-amylase activities were decreased from 8 days after seeding. Total free sugar contents were decreased during the germination period. There was continuous decline in the contents of sucrose and glucose until 8 days after seeding, but fructose and maltose content were gradually increased from the 5 days after seeding.

• Journal of Pharmacopuncture
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• v.19 no.4
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• pp.319-328
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• 2016

Objectives: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). Methods: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf ) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. Results: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. Conclusion: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.

• Journal of Pharmacopuncture
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• v.20 no.2
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• Polymer(Korea)
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• v.31 no.1
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• pp.14-19
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• 2007

Demineralized bone particle (DBP) has been used as one of the powerful inducers of bone and cartilage tissue specialization. In this study, we fabricated DBP/PLGA scaffold for tissue engineered disc regeneration. We manufactured dual-structured scaffold to compose inner cylinder and outer doughnut similar to nature disc tissue. The DBP/PLGA scaffold was characterized by porosity, wettability, and water uptake ability. We isolated and cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells from rabbit intervertebral disc. We seeded NP cells into the inner core of the hybrid scaffold and AF cells into the outer portion of it. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl) -2,5- diphenyltetrazolium -bromide (MTT) test. PLGA and PLGA/DBP scaffolds were implanted in subcutaneous of athymic nude mouse to observe the formation of disc-like tissue in vivo. And then we observed change of morphology and hematoxylin and eosin (H&E). Formation of disc-like tissue was better DBP/PLGA hybrid scaffold than control. Specially, we confirmed that scaffold impregnated 20 and 40% DBP affected to proliferation of disc cell and formation of disc-like tissue.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.3
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• pp.288-298
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• 1992

Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(＋)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(＋)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.155-162
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• 2005

The present study evaluated whether an exogenous antioxidants, taurine and $\alpha$-tocopherol, could, when added to the freezing extender, improve the post-thaw sperm characteristics, function, the level of reactive oxygen species (ROS) generation, and the level of lipid peroxidation (LPO) in frozen-thawed porcine semen. CASA (computer-aided sperm analysis), HOST (hypoos-motic swelling test), chemiluminescence using luminol and lucigenin and the detection of malondialdehyde for LPO was performed in frozen-thawed porcine sper-matozoa. The results obtained in these studies are as follows. While no beneficial effects of taurine and $\alpha$-tocopherol supplementation were visible in motility, viability, acrosome reaction, tail swelling patterns, and the generation of $O^{2-}$ of frozen-thawed porcine sper-matozoa, $H_{2}O_{2}$ was decreased by all treatments except taurine 50mM treatment. In conclusion the taurine and $\alpha$-tocopherol treatments during freezing reduced generation of reactive oxygen species and production of malondialdehyde in frozen-thawed porcine semen, and the ROS savangers may minimize various damages of spermatozoa during freezing.