Metabolizable protein (MP) supply and amino acid balance in the intestine were manipulated through selection of highly digestible rumen-undegradable protein (RUP) sources and protected methionine (Met) supplementation. Four ruminallycannulated, multiparous Holstein cows averaging 193${\pm}$13 days in milk were used in a 4${\times}$4 Latin square design to assess N utilization and milk production responses to changes in RUP level, post-ruminal RUP digestibility and protected Met supplementation. Treatments were A) 14.0% crude protein (CP), 8.0% rumen degradable protein (RDP) and 6.0% RUP of low intestinal digestibility (HiRUP-LoDRUP); B) 14.1% CP, 8.1% RDP and 6.0% RUP of high intestinal digestibility (HiRUP-HiDRUP); C) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility (LoRUP-HiDRUP), and D) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility plus rumen escape sources of Met (LoRUP-HiDRUP+Met). Experimental diets were formulated to have similar concentrations of RDP, net energy of lactation ($NE_L$), neutral detergent fiber (NDF), acid detergent fiber (ADF), calcium, phosphorus and ether extract using the NRC model (2001). Results showed that dry matter intake (DMI), production of milk fat and protein were similar among treatments. Milk production was similar for diet HiRUP-LoDRUP, HiRUP-HiDRUP and LoRUP-HiDRUP+Met, and significantly higher than diet LoRUP-HiDRUP. Milk fat and protein percentage were higher for cows receiving HiDRUP treatments, with the greatest increases in the diet LoRUP-HiDRUP+Met. There was no significant change in ruminal pH, $NH_3g-N$ and volatile fatty acid (VFA) concentration among all treatments. Apparent digestibility of dry matter (DM), CP, NDF and ADF and estimated bacterial CP synthesis were similar for all treatments. Nitrogen intakes, blood and milk urea-N concentrations were significantly higher for cows receiving HiRUP diets. Urine volume and total urinary N excretion were significantly lowered by LoRUP diets. Lowering dietary RUP level while supplementing the highly digestible RUP source with rumen escape sources of Met resulted in similar milk production, maximal milk fat and protein concentration and maximum N efficiency, indicating that post-ruminal digestibility of RUP and amino acid balance in the small intestine can be more important than total RUP supplementation.
Two experiments were conducted to evaluate nutritive value of cell mass from lysine production (CMLP) as a protein supplement for ruminants. In each experiment, animals were fed a diet containing 40% of forages and 60% of concentrates, mainly composed of rice straw and ground corn, respectively, to meet the maintenance requirements, and the diets were formulated to supply equal amounts of energy and nitrogen among treatments. In order to investigate the effect of CMLP on ruminal fermentation (Experiment 1), three Korean native goats weighing $26.1{\pm}1.4kg$ were allotted into individual cages with a $3{\times}3$ Latin square design. Each animal was fed one of three protein sources (CMLP, soybean meal (SBM), and urea). Rumen pH, bacterial and fungal counts, volatile fatty acid concentrations and acetate to propionate ratio were not significantly different among treatments. Concentration of propionate, however, was higher in SBM treatment (14.1 mM) than in CMLP (8.7 mM) or urea (9.3 mM) treatments. There was significantly more branch-chain volatile fatty acid production in CMLP (1.9 mM) and SBM (1.8 mM) treatments than in urea (1.3 mM) treatment. The number of protozoa was the highest in urea treatment, followed by CMLP and SBM treatment with significant differences. A metabolic trial (Experiment 2) was conducted to measure in vivo nutrient digestibility and nitrogen retention in Korean native goats fed CMLP and SBM. Two heavy ($35.0{\pm}1.2kg$) and two light ($25.0{\pm}0.9kg$) Korean native goats, caged individually, were used in this experiment. A heavy and a light animal were paired and supplemented with either CMLP or SBM. The animals fed CMLP showed a trend of lower total tract digestibility in all the nutrients measured; however, there was no statistical significance except for digestibility of ether extract. Nitrogen digestibility of CMLP was estimated to be about 7% units lower than that of SBM. There was a tendency for lower nitrogen retention in CMLP treatment (35.9%) compared to SBM treatment (42.3%). In summary, CMLP can be a good protein source for ruminant animals from nutritional and economic perspectives and may replace some, if not all, of SBM in a diet without losing nitrogen utilization efficiency. Further research is warranted for investigating the effect of CMLP fed with easily fermentable forage and the effective level of CMLP for replacing SBM.
Biodegradation of fat, oil, and grease (FOGs) plays an Important role in wastewater management and water pollution control. However, many industrial food-processing and food restaurants generate FOG-containing waste waters for which there Is no acceptable technology for their pretreatment. To solve these problems, this study evaluated the feasibility of effective FOG-degrading microorganisms on the biodegradation of olive oil and FOG-containing wastewater. Twenty-two strains capable of degrading FOGs were isolated from five FOG-contaminated sites for the evaluation of their FOG degradation capabilities. Among twenty-two strains tested, the lipase-producing Pseudomonas sp. strain D2D3 was selected for actual FOG wastewater treatment. Its biodegradability was performed at 3$0^{\circ}C$ and pH 8. The extent of FOG removal efficiency was varied for each FOG tested, being the highest for olive oil and animal fat (94.5% and 94.4%), and the lowest for safflower oil (62%). The addition of organic nitrogen sources such as yeast extract, soytone, and peptone enhanced the removal efficiency of FOGs, but the addition of the inorganic nitrogen nutrients such as $NH_4$Cl and $(NH_4)_2SO_4$ did not increase. The $KH_2PO_4$ sources in 0.25% to 0.5% concentrations showed more than 90% degradability. As a result, the main pathway for the oxidation of fatty acids results in the removal of two carbon atoms as acetyl-CoA with each reaction sequence: $\beta$-oxidation. Its lipase activity showed 38.5 U/g DCW using the optimal media after 9 h. Real wastewater and FOGs were used for determining the removal efficiency by using Pseudomonas sp. strain D2D3 bioadditive. The degradation by Pseudomonas sp. strain D2D3 was 41% higher than that of the naturally occurring bacteria. This result indicated that the use of isolated Pseudomonas sp. strain D2D3 in a bioaugmentating grease trap or other processes might possibly be sufficient to acclimate biological processes for degrading FOGs.
Objectives : The aim of this study was to evaluate the efficacy of Hwangryunhaedok-tang (HHT) and Gungangbuja-tang (GBT) on lipopolysaccharide (LPS)-induced mouse model of inflammation. HHT and GBT are one of the representative prescriptions of cold drug and one of the representative prescriptions of hot drug, respectively. For experimental evaluation of their efficacy, we investigated the anti-inflammatory effects of HHT and GBT on LPS-induced inflammation and the mechanisms of their action. Methods : ICR mice were given a HHT (50, 500 mg/kg), GBT (100, 1000 mg/kg) extract orally on three consecutive days. On the third day, they were administered LPS intraperitoneally (35 mg/kg), 1 h after the last sample administration. Blood and liver samples were taken 6 h after the LPS challenge. Cytokine expression and inflammation-related protein factor analyses were performed by Western blotting. Results : Oral administration of HHT significantly reduced pro-inflammatory cytokines, including interleukin (IL)-6, and interferon (IFN)-${\gamma}$ in the serum. While GBT inhibited an increase of IL-6, IFN-${\gamma}$ was not affected. Immunoblot analysis showed that LPS-induced NF-${\kappa}b$ activation was inhibited by GBT, meanwhile HHT only inhibited NF-${\kappa}b$ expression at high does (500 mg/kg). In addition, HHT and GBT inhibited LPS-induced phosphorylation of Erk1/2, Jnk and p38 MAPKs. GBT also significantly inhibited i-Nos and Cox-2 expression, and HHT inhibited only i-Nos expression. Conclusions : Both of HHT and GBT showed anti-inflammatory effects against LPS-induced endotoxemia. However, HHT significantly decreased inflammatory cytokine levels, such as IL-6 and IFN-${\gamma}$ more than GBT, while GBT significantly inhibited inflammatory proteins, including NF-${\kappa}b$, MAP Kinases, i-Nos and Cox-2, more than HHT. These results suggest that HHT and GBT regulate the different mechanisms of action and pathways, presumably by regulating cytokine levels (IL-6, IFN-${\gamma}$), NF-${\kappa}b$ activation, and several pro-inflammatory gene expression, although both of HHT and GBT have anti-inflammatory effects.
Bisphenol A (BPA) has been widely used as a monomer for production of epoxy resins and polycarbonate plastics. The annual production of BPA exceeds 640,000 metric tons in worldwide. BPA, a suspected phenolic endocrine disrupter, is moderately soluble and frequently detected in industrial wastewater. To date, HPLC and GC has been used for BPA analysis. However, HPLC and GC-analysis need high operation lost, experts, and an elaborate pre-treatment of samples, and is difficult to apply on-time and mass analysis. Therefore, simple, mass and rapid detection of BPA in environments is necessary. In the present study, spectrophotometric method of BPA quantification was developed. Based on blue-color product formation with BPA and ferric chloride/ferricyanide under the optimized conditions, the standard curve was acquired $({\lambda}_{750}=0.061\;BPA\;[{\mu}M]+0.07155,\;R^2=0.992)$. Using an established method, the BPA contents in the soil extract, and different water samples and living products, including disposable syringe, cup and plastic tube, were analyzed. The results suggested that the method is useful for BPA determination from different massive samples. Since the BPA metabolites, nontoxic 4-hydroxyacetophenone or 4-hydroxybenzaldehyde, did not form blue-color product, this method is also useful to screen a microorganism for BPA bioremediation.
Sclerotinia sclerotiorum, a plant pathogenic fungus, can cause serious yield and quality losses in the winter lettuce field. For biological control of S. sclerotiorum, soil-born microorganisms that inhibit the mycelia growth of S. sclerotiorum and Fusarium oxysporum were isolated from diseased soil. Among the isolates, bacterial isolate, GG95, which was identified as Bacillus subtilis according to the morphological, physiological characteristics and by 16S rRNA similarity, showed the highest level of inhibitory activity. The growth conditions for B. subtilis GG95 were optimized in TSB media (pH 7) by culturing at $28^{\circ}C$ for 24 hrs. Maltose or fructose and peptone were selected as the best carbon and nitrogen sources, respectively. Greenhouse experiment was performed to test effectiveness of B. subtilis GG95 in the control sclerotinia rot. Drench application ($1{\times}10^8cfu/mL$, 3 times) of the bacterial culture broth to lettuce showed an effectiveness value of 88%, suggesting that B. subtilis GG95 would be a promising biocontrol agent for control of sclerotinia rot.
In this study, we tried to establish the best method for fresh blueberry beverage production using enzyme treatment as well as low temperature extraction. During extraction of physiologically functional materials, we used low temperature to prevent nutritional loss by heat. In addition, we investigated optimal blueberry extraction conditions using various enzyme treatments (cellulase, pectinase, cellulase:pectinase (1:1) mixture) to increase extraction efficiency and reduce turbidity. A variety and ratio of enzymes, extraction temperature, extraction time, and shaking speed were considered for the best extraction efficiency rate. We observed high extraction efficiency rates of 85.72-86.55% and 87.06-87.93%, respectively, upon cellulase or pectinase treatment. In addition, a mixture of cellulase:pectinase (1:1) showed an extraction efficiency rate of 86.84-88.14%. The best extraction efficiency rate was observed when crude blueberry was treated at $45^{\circ}C$ (87.91%), for 3 h (87.88%), in a 90 rpm shaker (89.19%). Sugar content and acidity of blueberry extract were not affected by the various treatments. However, total phenolic compounds were detected upon pectinase treatment (18.62 mg/g). Only fructose and glucose as free sugars were found in all samples regardless of treatments and extraction conditions.
Kim, H.S.;Choi, S.W.;Yun, A.R.;Lee, S.E.;Shin, K.Y.;Choi, J.I.;Mun, J.H.
Journal of Biosystems Engineering
/
v.34
no.2
/
pp.127-132
/
2009
In the working population, muscle fatigue and musculoskeletal discomfort are common, which, in the case of insufficient recovery may lead to musculoskeletal pain. Workers suffering from musculoskeletal pains need to be rehabilitated for recovery. Isokinetic testing has been used in physical strengthening, rehabilitation and post-operative orthopedic surgery. Frequency analysis of electromyography (EMG) signals using the mean frequency (MNF) has been widely used to characterize muscle fatigue. During isokinetic contractions, EMG signals present strong nonstationarities. Hilbert-Haung transform (HHT) and autoregressive (AR) model have been known more suitable than Fourier or wavelet transform for nonstationary signals. Moreover, several analyses have been performed within each active phase during isokinetic contractions. Thus, the aims of this study were i) to determine which one was better suitable for the analysis of MNF between HHT and AR model during repetitive maximum isokinetic extensions and ii) to investigate whether the analysis could be repeated for sequential fixed epoch lengths. Seven healthy volunteers (five males and two females) performed isokinetic knee extensions at $60^{\circ}/s$ and $240^{\circ}/s$ until 50% of the maximum peak torque was reached. Surface EMG signals were recorded from the rectus femoris of the right thigh. An algorithm detecting the onset and offset of EMG signals was applied to extract each active phase of the muscle. Following the results, slopes from the least-square error linear regression of MNF values showed that muscle fatigue of all subjects occurred. The AR model is better suited than HHT for estimating MNF from nonstationary EMG signals during isokinetic knee extensions. Moreover, the linear regression can be extracted from MNF values calculated by sequential fixed epoch lengths (p> 0.0I).
Natural products have always been an attractive source in terms of novel anti-metastatic compounds which can hinder MMP expression and activity. Corydalis heterocarpa is a salt marsh plant found in the seashores throughout Korea. Its yellow flowers and spikes have been an ingredient in folk medicine to treat spasm and contractions. The present study assessed the potential of different solvent-based fractions from the crude extract of Corydalis heterocarpa (CHE), a halophyte with reported bioactivities, to suppress the PMA-induced MMP expression in human fibrosarcoma HT-1080 cells. The solvent fractions which were named after the solvent used for fractionation (n-hexane, 85% aqueous (aq.) methanol (MeOH), n-butanol (BuOH), and H2O were shown to inhibit the both elevated mRNA and protein expression levels of MMP-2 and MMP-9 and simultaneously relieved the suppression on the expression of the endogenous MMP inhibitors TIMP-1 and TIMP-2. Results indicated that the CHE fractions might intervene with the PMA-induced activation of the MAPK signaling which is the upstream activator of MMP overexpression. Among tested samples, 85% aq. MeOH and n-hexane fractions of CHE was determined to be the most active and future studies to isolate the bioactive substances responsible for the regulation of the MMP expression are, therefore, urged. In conclusion, C. heterocarpa was shown to be a potential source of anti-metastatic compounds and n-Hexane and MeOH fractions might yield lead molecules to develop novel MMP inhibitors.
Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.
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