The immunomodulatory effects of Aureohasidium pullulans SM-2001 exopolymers containing $\beta$-1,3/1,6-glucan were evaluated in cyclophosphamide (CPA)-treated mice. To induce immunosuppression, 150 and 110 mg/kg of CPA were intraperitoneally injected 3 days and 1 day, respectively, before beginning administration of the test material. Exopolymers were delivered subcutaneously or orally, four times, in a volume of 10 ml/kg at 12-h intervals beginning 24 h after the second CPA treatment. Changes in thymus and spleen weights, splenic amounts of tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1{\beta}$, and IL-10, and numbers of CD3+, CD4+, CD8+, and TNF-$\alpha+$ thymus and spleen cells were monitored in CPA-treated mice. As a result of CPA treatment, dramatic decreases in the number of CD3+, CD4+, CD8+, and TNF-$\alpha+$ cells were detected in the thymus and spleen, along with decreases in thymus and spleen weights. In addition, splenic TNF-$\alpha$, IL-$1{\beta}$, and IL-10 contents were also decreased on observation with flow cytometry. However, oral and subcutaneous treatments with exopolymers effectively reduced the immunosuppressive changes induced by CPA. Therefore, it is concluded that exopolymers of A. pullulans SM-2001 can effectively prevent immunosuppression through, at least partially, the recruitment of T cells and TNF-$\alpha+$ cells or enhancement of their activity, and can provide an effective component of prevention or treatment regimens for immunosuppression related to cancer, sepsis, and high-dose chemotherapy or radiotherapy.
Fermented medical herbs using Lactobacilli have attracted significant attention due to their enhanced biological activities. A traditional medicinal plant, Artemisia annua L., was fermented using a probiotic strain, L. plantarum SK3494. The strain was isolated from Artemisia princeps var. orientalis and molecularly identified through sequence similarities and phylogenetic tree analysis. The antioxidant activity of L. plantarum-fermented A. annua L. (LFA) was determined using the DPPH free radical scavenging assay. Cellular antioxidant activity of LFA was examined using the superoxide radical reduction assay in MAT-C cells. Total polyphenol contents (TPC) and flavonoid contents (TFC) of LFA were determined. The antibacterial activity of LFA against fish pathogens was also determined in this study. The viable cell number (9.38 log10 CFU/ml) and pH (4.1) results showed good adaptive ability of the selected strain during fermentation. LFA was found to have enhanced antioxidant activity compared to non-fermented A. annua L. (NFA) based on the DPPH assay. Cellular antioxidant activity was present in both LFA and NFA. After 24 hr and 48 hr of fermentation, the LFA also showed antibacterial activities against fish pathogens Photobacterium damselae subsp. damselae and Vibrio ichthyoenteri. These results suggest that L. plantarum-fermented A. annua L. may have potential as a feed additive in aquaculture.
Major objectives were to evaluate effects of three schemes of bST-supplementation of Holstein cows (142.8 mg/14 d, POSILAC) during the prepartum and/or postpartum periods through 63 d (${\pm}3d$) of lactation. Measures evaluated the potential of treatments to improve body weight (BW) and body condition score (BCS), provoke changes in plasma concentrations of somatotropin (ST) and IGF-I, and improve milk yield, milk composition (percentages of protein and fat, and somatic cell counts), and several calving variables. Multiparous Holstein cows were randomly assigned to a $2{\times}2$ factorial arrangement of treatments (TRT) to give four groups (I = no bST, n = 26; II = bST postpartum, n = 25; III = bST prepartum, n = 27; IV = bST prepartum and postpartum, n = 25). During the prepartum period, cows in groups I and II were not supplemented but those in groups III and IV were supplemented every 2-wk beginning 21 d before expected calving date through calving. During the first 63 DIM only cows in groups II and IV were supplemented with bST. From 64 DIM through the end of lactation cows in all groups were supplemented with the full lactation dose of bST (500 mg/14 d). The BW and BCS were recorded weekly throughout the prepartum and postpartum periods and every 2-wk beyond 70 DIM. Blood samples were collected 3-times a week for analyses of ST and IGF-I. Milk yields were recorded daily though 150 DIM. Prepartum supplementation of bST did not affect BW or BCS, but mean concentrations of ST were increased 12.2% and were 15.5% greater at calving. Overall, mean concentration of IGF-I was not affected by treatment but concentrations were greater at 1 and 2 wk before calving in bST-supplemented cows. During the first 63 DIM the BW and BCS were not affected by treatment. Significant effects of bST-supplementation were detected on concentrations of ST, IGF-I and on milk yield compared to non-supplemented cows in group I. Postpartum concentrations of ST were greater in bST-supplemented cows (TRT II and IV; +41.9 and 54.6%). However, concentrations of IGF-I were greater only in cows in group IV (+25.9%) during the postpartum period. Overall, the three bST-supplemented groups had greater actual milk yield than the control group (I) during the first 63 and 150 DIM. The actual milk yields during 63 and 150 DIM were 6.5 and 4.6 kg/d greater for cows in group IV than cows in group I and the 305-d ME milk yield also was 15.6% greater. No adverse effects of TRT were observed on calf birth weight, colostrum immunoglobulins, ease of calving or other measures evaluated.
Four hundred and fifty tilapias ($6.77{\pm}0.23$ g) were assigned randomly to six groups to evaluate the feasibility of the tested antibacterial peptides (ABPs) and oligosaccharides as substitutes for antibiotics. The control group was fed with a commercial tilapia diet; other five groups were fed with the same commercial diet supplemented with konjac glucomannan (KGLM), cluster bean galactomannan (CBGAM), and three animal intestinal ABPs derived from chicken, pig and rabbit at 100 mg/kg respectively. After 21 days of feeding, growth, disease resistance, and in vivo anti-adherence were determined. Furthermore, the inhibitory effect of tested agents on adhesion of Aeromonas veronii biovar sobria (A.vbs) strain BJCP-5 to tilapia enteric epithelia in vitro was assessed by cell-ELISA system. As a result, the tested agents supplemented at 100 mg/kg show significant benefit to tilapia growth and disease resistance (p<0.05), and the benefit may be correlated with their interfering in the contact of bacteria with host mucosal surface. Although none of the tested agents did inhibit the growth of BJCP-5 in tryptic soy broth at $100{\mu}g/ml$, all of them did inhibit the adhesion of A.vbs to tilapia enteric epithelia in vivo and in vitro. In vitro mimic assays show that three ABPs at low concentrations of $25{\mu}g/ml$ and $2.5{\mu}g/ml$ have the reciprocal dose-dependent anti-adherence effect. The inhibition of ABPs may be correlated with a cation bridging and/or receptor-ligand binding, but not with hydrophobicity. The KGLM and CBGAM inhibited the adherence of BJCP-5 to tilapia enteric epithelia with dose-dependent manner in vitro, and this may be through altering bacterial hydrophobicity and interfering with receptor-ligand binding. Our results indicate that the anti-adherence of the tested ABPs and oligosaccharides may be one of the mechanisms in promoting tilapia growth and resistance to A.vbs.
Gong, W.H.;Tang, Z.L.;Han, J.L.;Yang, S.L.;Wang, H.;Li, Y.;Li, K.
Asian-Australasian Journal of Animal Sciences
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v.21
no.11
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pp.1544-1550
/
2008
The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.
We examined effects of interleukin $1{\alpha}$ ($IL1{\alpha}$) and phorbol 12, 13 dibutyrate (PDB), an activator of protein kinase C, on mRNA for Prostaglandin H synthase (PGHS) and prostanoid production in cultured ovine meningeal fibroblasts. Immuno- and morphologically-identified fibroblasts were derived from cerebral cortex and white matter from fetal lambs (approximately 120 days gestation) and grown to confluence on glass coverslips in 12 well plates. Levels of prostaglandin $F_{2{\alpha}}$ and the stable hydrolysis product of prostacyclin (i.e., $6-keto-PGF_{1{\alpha}}$) were determined using enzyme immunoassay. Relative amounts of mRNA were determined by in situ hybridization using ovine cDNA for PGHS1. $IL1{\alpha}$ (10 ng/ml) increased mRNA levels over baseline by $62{\pm}19%$ (p<0.05) at 60 min., $37{\pm}12%$ (NS) at 120 min., and $36{\pm}18%$ (NS) at 240 min (n=12). Levels of $6-keto-PGF_{1{\alpha}}$ were $148{\pm}18%$ pg/ml during baseline, $246{\pm}41%$ pg/ml at 60 min., $248{\pm}40%$ pg/ml at 120 min., and $259{\pm}62%$ pg/ml at 240 min (all p<0.05) (n=12). $PGF_{2{\alpha}}$ was increased although it wasn't statistically significant. However, $IL1{\alpha}$ decreased $PGE_2$ level significantly (all p<0.05). PDB $(10^{-6}M)$ increased mRNA levels over baseline by $25{\pm}6%$ after 30 min., $40{\pm}6%$ after 60 min., and $20{\pm}8%$ after 90 min. (n=9) (all p<0.05). Levels of $6-keto-PGF_{1{\alpha}}$ were $200{\pm}43%$ pg/ml during baseline, $202{\pm}43%$ pg/ml after 30 min. (NS), $268{\pm}58%$ pg/ml after 60 min. (p<0.05), and $296{\pm}60%$ pg/ml after 90 min. (p<0.05) (n=9). Levels of $PGF_{2{\alpha}}$ were $178{\pm}26%$ pg/ml during baseline, $300{\pm}30%$ pg/ml after 30 min., $299{\pm}35%$ pg/ml after 60 min., and $355{\pm}32%$ pg/ml after 90 min (all p<0.05) (n=6). Actinomycin-D (1 mg/ml) prevented increases in mRNA, $6-keto-PGF_{1{\alpha}}$, and $PGF_{2{\alpha}}$ at 60 min. for both $IL1{\alpha}$ and PDB. We conclude that cerebral fibroblasts are avid producers of prostanoids, and that enhanced production of PGHS is responsible for augmented $PGF_{2{\alpha}}$ and prostacyclin production in the presence of an activator of protein kinase C and for decreased $PGE_2$ and increased prostacyclin production in the presence of $IL1{\alpha}$.
Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.
Purpose: Sclerostin, an inhibitor of Wnt/${\beta}$-catenin signaling, exerts negative effects on bone formation and contributes to periodontitis-induced alveolar bone loss. Recent studies have demonstrated that serum sclerostin levels are increased in diabetic patients and that sclerostin expression in alveolar bone is enhanced in a diabetic periodontitis model. However, the molecular mechanism of how sclerostin expression is enhanced in diabetic patients remains elusive. Therefore, in this study, the effect of hyperglycemia on the expression of sclerostin in osteoblast lineage cells was examined. Methods: C2C12 and MLO-Y4 cells were used in this study. In order to examine the effect of hyperglycemia, the glucose concentration in the culture medium was adjusted to a range of levels between 40 and 100 mM. Gene expression levels were examined by quantitative reverse transcription-polymerase chain reaction and Western blot assays. Top-Flash reporter was used to examine the transcriptional activity of the ${\beta}$-catenin/lymphoid enhanced factor/T-cell factor complex. Tumor necrosis factor-alpha ($TNF{\alpha}$) protein levels were examined with the enzyme-linked immunosorbent assay. The effect of reactive oxygen species on sclerostin expression was examined by treating cells with 1 mM $H_2O_2$ or 20 mM N-acetylcysteine. Results: The high glucose treatment increased the mRNA and protein levels of sclerostin. High glucose suppressed Wnt3a-induced Top-Flash reporter activity and the expression levels of osteoblast marker genes. High glucose increased reactive oxygen species production and $TNF{\alpha}$ expression levels. Treatment of cells with $H_2O_2$ also enhanced the expression levels of $TNF{\alpha}$ and sclerostin. In addition, N-acetylcysteine treatment or knockdown of $TNF{\alpha}$ attenuated high glucose-induced sclerostin expression. Conclusions: These results suggest that hyperglycemia increases sclerostin expression via the enhanced production of reactive oxygen species and $TNF{\alpha}$.
Cellular glass foam (CGF), the reprocessed glass, has a possibility to be used as a medium component in plug culture of horticultural crops due to the its excellent air and water permeability as comparable to perlite. An experiment was conducted to investigate growth of plug seedlings of Lycopersicum esculentum 'Segye' as influenced by irrigation frequency in various medium combinations of CGF (2.0-4.0 mm particle size) and peatmoss. Seeds were sown in 200-cell plug trays, filled with mixtures of CGF and peatmoss either at 33:67 or 25:75 (%. v/v) and were germinated on a fogged propagation bed. The irrigation frequencies used were one, two or three times per every two days. A commercial plug medium (Tosilee medium) was used as the control, and the irrigation frequency in the control was one time per day. Growth of seedlings, and medium pH and EC were measured at 33 days after sowing. The medium composition had little influence on overall growth of seedlings. Irrigation frequency very significant affected number of leaves, leaf area, chlorophyll concentration, fresh and dry weights of shoots and roots, and dry matter. Growth of seedlings was the greatest with the highest irrigation frequency in the 25% CGF+75% peatmoss mixture.
Journal of the Korea Organic Resources Recycling Association
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v.26
no.4
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pp.53-61
/
2018
Anaerobic digestion (AD) is one of the most widely used process that can convert the organic fraction of waste activated sludge (WAS) into biogas. However, most researched actual methane yields of anaerobic digester (AD) on lab scale is lower than theoretical ones. Bioelectrochemical, anaerobic digester was used to increase methane yield from waste activated sludge. The influence of anaerobic digestion sludge and raw sludge mixing ratio (3:7, 5:5) on methane yield and organic matter removal efficiency were explored. As a result, when the mixing ratio of bioelectrochemical anaerobic sludge was 5:5 compared with 3:7, the highest methane yields were 294.2 mL $CH_4/L$ (0.63 times increase) and 52.5% (7.5% increase), the bioelectrochemical anaerobic digester(5:5) was more stable in the pH, t otal alkalinity and VFAs, respectively. These results showed that the increase in the mixing ratio of anaerobic digestion sludge was found to be effective for maintaining the stable performance of bioelectrochemical anaerobic digester.
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