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In vivo Effects of Di-n-butyl Phthalate and Di-2-ethylhexyl Phthalate on the Nonspecific Defense Mechanism of the Bagrid Catfish, Pseudobagrus fulvidraco

  • Masroor Fatima;Jee Jung-Hoon;Keum Yoo-Hwa;Kang Ju-Chan
    • Fisheries and Aquatic Sciences
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    • v.9 no.1
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    • pp.14-21
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    • 2006
  • The aim of this study was to investigate in vivo toxicity and effects of two phthalate esters (PEs), di-n-butylphthalate (DBP) and di-2-ethylhexyl phthalate (DEHP), on the immune system of the bagrid catfish, Pseudobagrus fulvidraco. Groups of experimental fish were subjected to daily intraperitoneal injections of 300 or 1000 mg $kg^{-1}$ of DBP or DEHP for 3 days, and the cellularity and functional activity of phagocytes were measured in the spleen and pronephros (head kidney). The number of spleen leukocyte cells increased significantly (p<0.05) in response to low and high doses of DEHP and DBP, respectively; however, the cellularity of the pronephros was more susceptible to higher dose of DEHP than DBP. Nonspecific immunity, as determined by the phagocytic index (PI) and phagocytic capacity (PC), was significantly depressed by DEHP at 1000 $1000mg\;kg^{-1}\;day^{-1}$ in the pronephros at 3 days after injection. Furthermore, significantly (p<0.05) increased levels of serum glutamic oxaloacetate transaminase (GOT) and glutamic pyruvate transaminase (GPT) indicated marked hepatic dysfunction in immunosuppressed fish. Treated fish showed a significant reduction in total serum protein but no significant alteration in lysozyme activity. These results demonstrate the sensitivity of the fish immune response for predicting PE-induced immunotoxicity.

Gelam and Nenas Honeys Inhibit Proliferation of HT 29 Colon Cancer Cells by Inducing DNA Damage and Apoptosis while Suppressing Inflammation

  • Wen, Christinal Teh Pey;Hussein, Saba Zuhair;Abdullah, Shailah;Karim, Norwahidah Abdul;Makpol, Suzana;Yusof, Yasmin Anum Mohd
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1605-1610
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    • 2012
  • Gelam and Nenas monofloral honeys were investigated in this study for their chemopreventive effects against HT 29 colon cancer cells. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolim) assays showed more effective inhibition of colon cancer cells proliferation by Gelam honey with $IC_{50}$ values of 39.0 mg/ml and 85.5 mg/ml respectively after 24 hours of treatment. Alkali comet assays revealed both honeys increased DNA damage significantly in a dose dependent manner. In addition, annexin V-FITC/PI flow cytometry demonstrated that at $IC_{50}$ concentrations and above, both Gelam and Nenas honeys induced apoptosis significantlyat values higher than for necrosis (p<0.05). Measurement of prostaglandin $E_2$ ($PGE_2$) confirmed that Gelam and Nenas honeys reduced its production in $H_2O_2$ inflammation-induced colon cancer cells. In conclusion, our study indicated and confirmed that both Gelam and Nenas honeys are capable of suppressing the growth of HT 29 colon cancer cells by inducing apoptosis and suppressing inflammation.

Improvement of blood glucose control in type 2 diabetic db/db mice using Platycodon grandiflorum seed extract (도라지 종자 추출물의 처리가 제2형 당뇨 db/db 마우스의 혈당개선에 미치는 효과)

  • Kim, Tae Yeong;Kim, Seok Joong;Imm, Jee-Young
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.81-88
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    • 2020
  • The biological activities of Platycodon grandiflorum (PG) root extracts have been studied intensively, whereas there are limited number of studies on PG seed extract (PGSE). PGSE was prepared by ethanol extraction, and its antidiabetic effect was evaluated in mice with type 2 diabetes (C57BLKS/J-db/db). Results indicated that the administration of high-dose PGSE (600 mg/kg, wb) significantly stabilized the blood glucose levels, as evidenced by the results of the oral glucose tolerance test. Mice treated with high-dose PGSE exhibited significantly lower serum hemoglobin A1c, insulin, and leptin levels after eight weeks of feeding trial (p<0.05). High-dose PGSE administration significantly improved glucose uptake in the femoral muscle of db/db mice by activating both IRS-1/PI3K/AKT/AS160 and AMPK phosphorylation pathways. GLUT4 translocation from the cytosol to the plasma membrane increased 1.7-fold in the PGSE high-dose group. These results suggest that PGSE has potential for development as an antidiabetic agent.

Effects of Musculoskeletal Transplant Foundation on Bone Formation in Human Fetal Osteoblasts (사람태아골모세포에 대한 근골격이식재의 골형성 유도에 관한 효과)

  • Park, Jae-young;Pi, Sung-Hee;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.449-459
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    • 2006
  • DFDBA(Decalcified freeze-dried bone allograft) is one of the allograft materials for periodontal bone regeneration. DFDBA provides an osteoconductive surface and osteoinductive factors. Therefore, DFDBA have been used successfully to regenerate the attachment apparatus during periodontal treatment. But recent studies was reported that wide variations in commercial bone bank preparations of DFDBA do exist, including the ability to induce new bone formation. DFDBA was experimental materials that was recovered, processed, tested, shipped and invoiced through Musculoskeletal Transplant Foundation. MTF(Musculoskeletal Transplant Foundation) is the world largest, non-profit, AATB(American Association of Tissue Banks) accredited tissue bank. The objective of this study was to determine the effects of serial dilutions of a DFDBA on human fetal osteoblastic cell proliferation and their potential to form and mineralize bone nodules. Human fetal osteoblastic cell line(hFOB 1.19) was cultured with DMEM and SSE($1{\mu}g/m{\ell}$,$10{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $1mg/m{\ell}$) at $34^{\circ}C$ with 5% CO2 in 100% humidity. Cell proliferation was significantly increased at $1mg/m{\ell}$, $100{\mu}g$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDBA after 5 days incubation (p<0.05). Alkaline Phosphatase(ALP) level was significantly increased in $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDABA(p<0.05). A quantified calcium accumulation was significantly increased at $1ng/m{\ell}$, $10ng/m{\ell}$ of MTF(p<0.05). These results indicated that DFDBA has an inductive effect on bone formation in vitro.

Field Sanitation and Foliar Application of Streptomyces padanus PMS-702 for the Control of Rice Sheath Blight

  • Yang, Chia-Jung;Huang, Tzu-Pi;Huang, Jenn-Wen
    • The Plant Pathology Journal
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    • v.37 no.1
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    • pp.57-71
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    • 2021
  • Rice sheath blight (ShB), caused by Rhizoctonia solani Kühn AG1-IA, is one of the destructive rice diseases worldwide. The aims of this study were to develop biocontrol strategies focusing on field sanitation and foliar application with a biocontrol agent for ShB management. Streptomyces padanus PMS-702 showed a great antagonistic activity against R. solani. Fungichromin produced by S. padanus PMS-702, at 3.07 mg/l inhibited 50% mycelial growth, caused leakage of cytoplasm, and inhibited the formation of infection structures of R. solani. Fungichromin could reach to 802 mg/l when S. padanus PMS-702 was cultured in MACC broth for 6 days. Addition of 0.5% S. padanus PMS-702 broth into soil decreased the survival rate of the pathogen compared to the control. Soil amended with 0.5% S. padanus broth and 0.5% tea seed pomace resulted in the death of R. solani mycelia in the infested rice straws, and the germination of sclerotia was inhibited 21 days after treatment. Greenhouse trials revealed that S. padanus cultured in soybean meal-glucose (SMGC-2) medium after mixing with different surfactants could enhance its efficacy for inhibiting the pathogen. Of six surfactants tested, the addition of 2% tea saponin was the most effective in suppressing the pathogen. S. padanus broth after being fermented in SMGC-2, mixed with 2% tea saponin, diluted 100 fold, and sprayed onto rice plants significantly reduced ShB disease severity. Thus, S. padanus PMS-702 is an effective biocontrol agent. The efficacy of S. padanus PMS-702 for disease control could be improved through formulation.

Study of the Effect and Underlying Mechanism of Clove Extract on Monosodium Iodoacetate-Induced Osteoarthritis in Rats (정향(丁香) 추출물이 골관절염 흰쥐에 미치는 효과 및 기전 연구)

  • Jin A Lee;Min Ju Kim;Seong-Wook Seo;Mi-Rae Shin
    • The Journal of Internal Korean Medicine
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    • v.43 no.6
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    • pp.1089-1104
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    • 2022
  • Objective: The aim of this study was to identify the efficacy and underlying mechanism of cloves as an osteoarthritis (OA) treatment in a monosodium iodoacetate (MIA)-induced rat OA model. Osteoarthritis (OA) is nowadays one of the most prevalent degenerative joint diseases. Methods: Sprague-Dawley rats treated with MIA (50 μL; 80 mg/mL) were used as in vivo OA models. Cloves (100 and 200 mg/kg b.w.) were administered orally once daily for 2 weeks from 7 days after MIA injection. Changes in hindpaw weight distribution (HWD) were measured as a joint discomfort index. Activation markers related to inflammatory responses and cartilage degeneration in the right knee joints were evaluated by serum analysis and western blotting. Results: HWD decreased in the MIA control group but showed a dose-dependent elevation after clove treatment. Clove treatment inhibited inflammatory factors by PI3K/Akt/NF-κB signaling pathways, while also activating antioxidant factors through Sirt1/AMPK signaling pathways. Clove treatment also suppressed matrix metalloproteinase (MMP) overexpression and significantly increased the levels of tissue inhibitors of metalloproteinases (TIMPs). Conclusions: Treatment with cloves effectively reversed MIA-induced effects. Therefore, clove treatment could have the potential to protect against or treat OA.

Studies on the Folic Acid and Methionine Requirements for Young Broiler Chicks Including New Analytical Methods for Folic Acid in Poultry Feedstuffs (가금 원료사료의 새로운 엽산 분석방법과 어린 육계의 엽산과 메티오닌의 요구량에 관한 연구)

  • 류경선;박강희;신원집
    • Korean Journal of Poultry Science
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    • v.22 no.3
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    • pp.179-188
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    • 1995
  • In Experiment 1, microbial assays were conducted on 57 feed ingredient samples to determine the content of total folic acid using Lactobacillus casei(ATCC 7469). Folic acid contents of feed samples pretreated with conjugase, ${\alpha}$-amylase, and a mixture of protease(Pronase)were corn, 09${\pm}$1.18($\pi$g${\pm}$SD); fish meal, 23.05${\pm}$1.27; milo, 29.34${\pm}$0.55; bakery meal, 25.80${\pm}$6.93; meat and bone meal, 56.76${\pm}$4.97; wheat middlings, 85.14${\pm}$2.56; and soybean meal, 193.97${\pm}$3.98. Experiments 2 and 3 were conducted to determine the effects of dietary supplemental folic acid and methionine on the performance of starting broiler chicks for 18 days. Four levels of dietary folic acid(0.24. 0.54,1.14 and 2.34mg/kg) and four levels of dietary methionine(0.45, 0.53,0.61, and 0.69%) were fed in a factorial design. The basal diet was based on corn, isolated soybean protein, meat and bone meal, and fish meal. It contained adequate amounts of all nutrients except methionine and folic acid in both experiments. Increased growth rate was observed in chicks fed the basal diet supplemented with either folic acid or methionine. Total dietary folic acid and methionine plus cysteine requirements for optimum growth were estimated to be 1.80 mg/kg and 0.89% in Experiment 2, and 1.47 mg/kg and 0.91% in Experiment 3, respectively. There were interactions between dietary folic acid and methionine on weight gain in both experiments. Chicks fed diets containing 2.34 mg folic acid /kg tended to display slow growth rate in both experiments. There was a significant linear feed conversion response to folic acid in Experiment 2, and a significant quadratic feed conversion resuonse to methionine in Experiment 3. There were both linear and quadratic liver folic acid responses to dietary folic acid in both experiments. There was no indication that dietary methionine had any effect on liver folic acid content. The incidence of tibial dyschondroplasia increased with increasing supplemental methionine, but were no significant differences detected at 5% level.

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Thermal Stability of Israeli Carp Actomyosin and Its Protection by Chemical Additives (이스라엘 잉어 Actomyosin의 열안정성과 그 보호)

  • NAM Taek-Jeong;CHOI Yeung-Joon;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.271-279
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    • 1984
  • Effects of temperature and additives on the stability of actomyosin extracted from skeletal muscle of Israeli carp, Cyprinus carpio nudus, were studied by analyzing free SH-group, ATP-sensitivity and Ca-ATPase activity. The used additives were sucrose, sorbitol, Na-glutamate and L-cysteine. Furthermore, the denaturation constant($K_D$), protective effect(${\Delta}E/M$) and the other thermo-dynamic parameters on protein denaturation are systematically discussed. The actomyosin showed $4.12{\sim}4.68 mg/ml$ in protein concentration, $2.63{\sim}2.93\%$ in ribonucleic acid to the protein, $1:2.20{\sim}2.63$ in the binding ratio of myosin and actin, $4.33{\sim}5.26\%$ in fat content, 109.78 in ATP-sonsitivity, $0.159{\sim}0.201\;{\mu}M-Pi/min/mg-protein$ in Ca-ATPase activity and $3.3{\sim}3.4M/10^5$g-protein in free SH-group content. The first-order rate plots were obtained on the decrease of Ca-ATPase activity and ATP-sensitivity with an increase in temperature, while the free SH-group was increased to $60^{\circ}C$ and decreased rapidly above the temperature. The half-life of Ca-ATPase activity on the actomyosin Ca-ATPase was 280 min at $12^{\circ}C$, 125 min at $20^{\circ}C$, 55 min at $30^{\circ}C$ and 13 min at $40^{\circ}C$, and activation energy, activation enthalpy, activation entropy and free energy of the proteins at $20^{\circ}C$ wene 5,395 cal/mole, 4,814 cal/mole, -40.42 e.u. and 17,626 cal/mole, respectively. The protective effect of the additives on the actomyosin Ca-ATPase showed that the most effective material is $3\%$ sorbitol and followed in the order of $8\%$ Na-glutamate, $1\%$ sucrose and $1\%$ L-cysteine. The actomyosin was more stable at $-30^{\circ}C$ than at $0^{\circ}C$ and $-20^{\circ}C$. and when the additives were used in the low temperature storage, $8\%$ Na-glutamate was the most effective. $3\%$ sorbitol, $1\%$ sucrose and $1\%$ L-cysteine was to become lower in the order.

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Distinct Pro-Apoptotic Properties of Zhejiang Saffron against Human Lung Cancer Via a Caspase-8-9-3 Cascade

  • Liu, Dan-Dan;Ye, Yi-Lu;Zhang, Jing;Xu, Jia-Ni;Qian, Xiao-Dong;Zhang, Qi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6075-6080
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    • 2014
  • Lung cancer is the leading cause of cancer-related death worldwide. Here we investigated the antitumor effect and mechanism of Zhejiang (Huzhou and Jiande) saffron against lung cancer cell lines, A549 and H446. Using high performance liquid chromatography (HPLC), the contents of crocin I and II were determined. In vitro, MTT assay and annexin-V FITC/PI staining showed cell proliferation activity and apoptosis to be changed in a dose- and time-dependent manner. The inhibition effect of Jiande saffron was the strongest. In vivo, when mice were orally administered saffron extracts at dose of 100mg/kg/d for 28 days, xenograft tumor size was reduced, and ELISA and Western blotting analysis of caspase-3, -8 and -9 exhibited stronger expression and activity than in the control. In summary, saffron from Zhejiang has significant antitumor effects in vitro and in vivo through caspase-8-caspase-9-caspase-3 mediated cell apoptosis. It thus appears to have more potential as a therapeutic agent.

Purification and Characterization of a Thermostable Alkaline Phosphatase Produced by Thermus caldophilus GK24

  • Kim, You-Jin;Park, Tae-Shin;Kim, Hyun-Kyu;Kwon, Suk-Tae
    • BMB Reports
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    • v.30 no.4
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    • pp.262-268
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    • 1997
  • The thermophilic and thermostable alkaline phosphatase was purified to near homogeneity from the osmotic lysis of Thermus caldophilus GK24, The purified enzyme had an apparent molecular mass of 108, 000 Da and consisted of two subunits of 54,000 Da. lsoelectric-focusing analysis of the purified enzyme showed a pi of 7.3. The enzyme contained two Cys residues, and its amino acids composition was quite different from that of Thermus aquaticus YT-1 alkaline phosphatase and Escherichia coli alkaline phosphatase, The optimum pH and temperature of the enzyme were 11.0-11.5 and $80^{\circ}C$ respectively. The enzyme was stable in the pH range of 9.0-12.0 at $25^{\circ}C$ for 36 h. and the half-life at $80^{\circ}C$ (pH 11.0) was 6 h. The enzyme was activated by $MgCl_2$ and inhibited by EDTA. With ${\rho}-nitrophenyl\;phosphate\;({\rho}NPP)$ as the substrate, the enzyme had a Michaelis constant $(K_m) $of $3.6{\times}10^{-5}M$, The enzyme preferentially hydrolyzed the phosphomonoester bond of AMP in ribonucleotides and glycerophosphate.

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