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Enhanced γ-aminobutyric acid and sialic acid in fermented deer antler velvet and immune promoting effects

  • Yoo, Jiseon;Lee, Juyeon;Zhang, Ming;Mun, Daye;Kang, Minkyoung;Yun, Bohyun;Kim, Yong-An;Kim, Sooah;Oh, Sangnam
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.166-182
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    • 2022
  • Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

Efficacy evaluation of novel organic iron complexes in laying hens: effects on laying performance, egg quality, egg iron content, and blood biochemical parameters

  • Jiuai Cao;Jiaming Zhu;Qin Zhou;Luyuan Zhao;Chenhao Zou;Yanshan Guo;Brian Curtin;Fei Ji;Bing Liu;Dongyou Yu
    • Animal Bioscience
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    • v.36 no.3
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    • pp.498-505
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    • 2023
  • Objective: This study was conducted to determine the optimal dose of novel iron amino acid complexes (Fe-Lys-Glu) by measuring laying performance, egg quality, egg iron (Fe) concentrations, and blood biochemical parameters in laying hens. Methods: A total of 1,260 18-week-old healthy Beijing White laying hens were randomly divided into 7 groups with 12 replicates of 15 birds each. After a 2-wk acclimation to the basal diet, hens were fed diets supplemented with 0 (negative control, the analyzed innate iron content was 75.06 mg/kg), 15, 30, 45, 60, and 75 mg Fe/kg as Fe-Lys-Glu or 45 mg Fe/kg from FeSO4 (positive control) for 24 wk. Results: Results showed that compared with the negative and positive control groups, dietary supplementation with 30 to 75 mg Fe/kg from Fe-Lys-Glu significantly (linear and quadratic, p<0.05) increased the laying rate (LR) and average daily egg weight (ADEW); hens administered 45 to 75 mg Fe/kg as Fe-Lys-Glu showed a remarkable (linear, p<0.05) decrease in feed conversion ratio. There were no significant differences among all groups in egg quality. The iron concentrations in egg yolk and serum were elevated by increasing Fe-Lys-Glu levels, and the highest iron content was found in 75 mg Fe/kg group. In addition, hens fed 45 mg Fe/kg from Fe-Lys-Glu had (linear and quadratic, p<0.05) higher yolk Fe contents than that with the same dosage of FeSO4 supplementation. The red blood cell (RBC) count and hemoglobin content (linear and quadratic, p<0.05) increased obviously in the groups fed with 30 to 75 mg Fe/kg as Fe-Lys-Glu in comparison with the control group. Fe-Lys-Glu supplementation also (linear and quadratic, p<0.05) enhanced the activity of copper/zinc-superoxide dismutase (Cu/Zn-SOD) in serum, as a result, the serum malonaldehyde content (linear and quadratic, p<0.05) decreased in hens received 60 to 75 mg Fe/kg as Fe-Lys-Glu. Conclusion: Supplementation Fe-Lys-Glu in laying hens could substitute for FeSO4 and the optimal additive levels of Fe-Lys-Glu are 45 mg Fe/kg in layers diets based on the quadratic regression analysis of LR, ADEW, RBC, and Cu/Zn-SOD.

Anti-inflammatory Effects of Houttuynia cordata and Lespedeza cuneata on Lipopolysaccharide-stimulated RAW264.7 Cells (마우스 대식세포 RAW264.7에서 어성초와 야관문의 항염증 효과)

  • Jeong Tae Kim;Chungwook Chung;Seong Ik Park;Man Hyo Lee;Joong Hee Roh;Ho Yong Sohn;Jong Sik Kim
    • Journal of Life Science
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    • v.33 no.1
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    • pp.73-81
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    • 2023
  • In the present study, we prepared hot water extracts and the subsequent organic solvent fractions of methanol extracts of Houttuynia cordata (HC) and Lespedeza cuneata (LC), and investigated their anti-inflammatory effects on lipopolysaccharide-stimulated RAW264.7 cells. Among the treated samples, hexane, chloroform, and ethyl-acetate fractions of HC and LC inhibited nitric oxide (NO) production in a dose-dependent manner, and decreased inducible nitric oxide synthase (iNOS) protein expression. And, we analyzed the flavonoid contents of the ethyl-acetate fraction of HC and LC, and chose apigenin for the further experiments because apigenin was one of flavonoids commonly found in HC and LC. Apigenin dramatically inhibited NO production in a dose-dependent manner without affecting cell viability and decreased iNOS and cyclooxygenase-2 (COX-2) expression. In addition, apigenin suppressed the phosphorylation of p38 and Jun N-terminal kinase (JNK) indicating that apigenin exerts anti-inflammatory activity via the mitogen-activated protein kinase (MAPK) signaling pathway. Subsequently, we conducted RNA-sequencing analysis to detect differentially expressed genes upon apigenin treatment. Among the down-regulated genes, four cytokine genes (interleukin (IL)-1α, IL-1β, IL-6, and colony stimulating factor 2 (CSF2)) were selected for the further analysis, and the reduction of their expression by apigenin was confirmed with quantitative real-time polymerase chain reaction. Overall, our results suggest that Houttuynia cordata and Lespedeza cuneata have the anti-inflammatory effects and apigenin can be the one of key molecules responsible for their anti-inflammatory activities.

Antithrombosis, Antidiabetes, and Antioxidant Activities of Houttuynia cordata (어성초의 항혈전, 항당뇨 및 항산화 활성)

  • Yun-Jin, Lee;Deok-Gyeong, Kang;Jong Sik, Kim;Man-Hyo, Lee;Ho-Yong, Sohn
    • Journal of Life Science
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    • v.33 no.1
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    • pp.43-49
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    • 2023
  • Houttuynia cordata belongs to the Saururacease family and its leaves, stems, and roots have been used as oriental medicines to treat pneumonia, acute or chronic bronchitis, enteritis, and abscesses and to remove extravasated blood. Recently, the antioxidant, anti-inflammation, antibacterial, and anti-proliferation activities and protection abilities of H. cordata against liver and neuron cell damage have been reported. In this study, ethanol extract and its solvent fractions (fractions of hexane, ethyl acetate, butanol, and water residue) were prepared, and their antithrombosis, antidiabetes, antioxidant, and hemolysis activities were evaluated. The ethyl-acetate fraction of H. cordata (EF-HC) showed the highest polyphenol and flavonoids contents among the fractions and exhibited strong antithrombosis and antioxidant activities. The EF-HC at 5 mg/ml showed 2.09-folds of thrombin time, 2.19-folds of prothrombin time, and 1.69-folds of activated partial thromboplastin time compared to the their solvent control and 30.9, 19.9, and 49.6 ㎍/ml of RC50 against DPPH, ABTS, and nitrite radicals, respectively. Furthermore, the EF-HC did not show any hemolytic activity up to 1 mg/ml, whereas the hexane fraction of H. cordata showed 55% hemolysis at 1 mg/ml. This is the first report of the antithrombosis activity of H. cordata. Our results suggest that quercitirin, hyperoside, orientin, and isoquercitrin in EF-HC are related to its antithrombosis and antioxidant activities and that the EF-HC could be developed as a promising antithrombosis agent.

Does supplementing laying hen diets with a herb mixture mitigate the negative impacts of excessive inclusion of extruded flaxseed?

  • Hossein Hosseini;Noah Esmaeili;Aref Sepehr;Mahyar Zare;Artur Rombenso;Raied Badierah;Elrashdy M. Redwan
    • Animal Bioscience
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    • v.36 no.4
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    • pp.629-641
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    • 2023
  • Objective: This study investigated the effects of extruded flaxseed with and without herbs mixture on egg performance, yolk fatty acids (FAs), lipid components, blood biochemistry, serological enzymes, antioxidants, and immune system of Hy-Line W-36 hens for nine weeks. Methods: Two hundred forty laying hens were randomly distributed to eight treatments, resulting in six replicates with five hens. Graded levels of dietary extruded flaxseed (0, 90, 180, and 270 g/kg) with and without herbs mixture (24 g/kg: garlic, ginger, green tea, and turmeric 6 g/kg each) were designed as treatments. Results: The two-way analysis of variance indicated that hens fed herbs mixture had a higher value of egg production, yolk high-density lipoprotein (HDL), superoxide dismutase, glutathione peroxidase, and white blood cell and lower contents of yolk cholesterol, glucose, and blood low-density lipoprotein than those fed diets without herb mixtures (p<0.05). The Flx27 (270 g/kg flaxseed) (153.5 g/kg n-3 FAs) and Flx27+H (270 g/kg flaxseed plus 24 g/kg herbs mixture) (150.5 g/kg n-3 FAs) groups were the most promising treatments in terms of yolk n-3 FAs content. In-teraction effect (herbs- flaxseed) for blood cholesterol, HDL, malondialdehyde, glutaredoxin, alanine transaminase, (ALT), aspartate transaminase (AST), haemoglobin and immune parameters was significant (p<0.05). The results showed layers fed herbs mixture (Flx9+H, Flx18+H, and Flx27+H) had a better value of total antibody, immunoglobulin M, immunoglobulin G, ALT, AST, and blood HDL as compared with representative flaxseed levels without herbs. Conclusion: High inclusion levels of extruded flaxseed (270 g/kg) without herbs to enrich eggs with n-3 appears to impair the antioxidant system, immunohematological parameters, and sero-logical enzymes. Interestingly, the herbs mixture supplementation corrected those effects. Therefore, feeding layers with flaxseed-rich diets (270 g/kg) and herbs mixture can be a promising strategy to enrich eggs with n-3 FAs.

The responsibility of C-terminal domain in the thermolabile haemolysin activity of Vibrio parahaemolyticus and inhibition treatments by Phellinus sp. extracts

  • Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.204-215
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    • 2023
  • The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

Neuroprotective Effect of Yukul-tang against the Oxidative Stress (육울탕(六鬱湯)의 산화적 스트레스에 대한 뇌세포 보호효과)

  • Jung, Sun-Hyung;Lee, Jin-Moo;Lee, Chang-Hoon;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.15-30
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    • 2009
  • Purpose: In this rapidly aging society, the research and development of traditional oriental medicine treatment is one of the critical factors to protect the increasing neuro-degenerative disorders. In this study, we wanted to verify the effect of Yukul-tang (YUT) on neuro-degenerative disease model by assessing the antioxidant and anti-inflammation effects. Methods: To assess the antioxidant effects of YUT, we carried out DPPH radical and ABTS radical scavenging assays and determined the total polyphenolic contents in YUT. To evaluate the neuro-protective effects of YUT, we performed the MTT and ROS assays and TH immunohistochemistry, NO and TNF-${\alpha}$ assays in SH-SY5Y or mesencephalic dopaminergic neurons damaged by 6-OHDA. Results: The treatment of YUT showed eliminating effects on DPPH radical and ABTS radical. it showed deterring effects on ROS, NO and TNF-${\alpha}$ and protecting effects on TH-positive cell in SH-SY5Y cells or mesencephalic dopaminergic neurons. Especially in the case of the treatment of YUT with 0.2ug/mL + 6-OHDA 10uM, the protective effect on dopaminergic neurons was most outstanding. Conclusion: In this study, we have demonstrated that YUT has an antioxidant effect and a neuro-protective effect on neuro-degenerative disease model caused by neurotoxin such as 6-OHDA. The results of our present study suggest that YUT can be useful agent to prevent and to treat neuro-degenerative diseases.

The Effect of Dangguijakyak-san on Wound Healing (당귀작약산의 창상 회복에 대한 효과)

  • Yun-Jin Lee;Chang-Hoon Woo;Young-Jun Kim;Hyeon-Ji Kim;Hee-Duk An
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.3
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    • pp.47-65
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    • 2023
  • Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.

Fabrication and Electrochemical Characterization of Carbon Fluoride-based Lithium-Ion Primary Batteries with Improved Rate Performance Using Oxygen Plasma (산소 플라즈마를 이용하여 율속 성능이 개선된 불화탄소 기반 리튬 일차전지의 제조 및 전기 화학적 특성)

  • Seoyeong Cheon;Naeun Ha;Chaehun Lim;Seongjae Myeong;In Woo Lee;Young-Seak Lee
    • Applied Chemistry for Engineering
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    • v.34 no.5
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    • pp.534-540
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    • 2023
  • The high-rate performance is limited by several factors, such as polarization generation, low electrical conductivity, low surface energy, and low electrolyte permeability of CFX, which is widely used as a cathode active material in the lithium primary battery. Therefore, in this study, we aimed to improve the battery performance by using carbon fluoride modified by surface treatment using oxygen plasma as a cathode for lithium primary batteries. Through XPS and XRD analysis, changes in the surface chemical characteristics and crystal structure of CFX modified by oxygen plasma treatment were analyzed, and accordingly, the electrochemical characteristics of lithium-ion primary batteries were analyzed and discussed. As a result, the highest number of semi-ionic C-F bonds were formed under the oxygen plasma treatment condition (7.5 minutes) with the lowest fluorine to carbon (F/C) ratio. In addition, the primary cell prepared under this condition using carbon fluoride as the active material of the cathode showed the highest 3 F/C(3 C rate-performance) rate-performance and maintained a relatively high capacity (550 mAh/g) even at high rates. In this study, it was possible to produce lithium primary batteries with high-rate performance by adjusting the fluorine contents of carbon fluoride and the type of carbon-fluorine bonding through oxygen plasma treatment.

The Photoprotective Effects of Gynura Procumbens Against Photoaging: A Comparative Study Based on Extraction Methods (추출방법에 따른 명월초(Gynura Procumbens)의 광노화 완화 효과)

  • So Yeon Han;Da Yoon Lee;Ji Sun Moon;Hye Won Lee;Jae Hun Kim;Jae Ho Park
    • Korean Journal of Plant Resources
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    • v.37 no.2
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    • pp.101-109
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    • 2024
  • In this study, we investigated the antioxidant effects and active ingredients of Gynura Procumbens extracts obtained through various extraction methods for the development of natural-based cosmetics and pharmaceutical materials. The contents of compounds, total flavonoids, chlorogenic acid, and caffeic acid were compared at different concentrations, revealing the highest content of active ingredients in the 100% ethanol extract. Antioxidant activity assessed by DPPH and ABTS radical scavenging assays showed a concentration-dependent increase in antioxidant activity with the ethanol concentration. Additionally, we validated the DNA damage inhibition and anti-inflammatory effects of Gynura Procumbens extracts through UVB irradiation on Hs68 cell models. The 100% ethanol extract demonstrated significant inhibition of the expression of p-p53, γ-H2AX, iNOS, and COX-2 induced by UVB, indicating its potential in alleviating photodamage effects. Consequently, the efficient extraction of Gynura Procumbens for skin functional material development was confirmed, suggesting the suitability of ethanol or alcohol-based solvents.