Hot-water extract, Fr. 1, of Phellinus linteus mycelium was fractionated into Fr. 2, 3, 4, and 5 by the difference of solubility in ethanol. The polysaccharide fractions were studied for their immunostimulating activity on in vitro T-independent polyc1onal antibody response to trinitrophenyl-haptened SRBC (sheep red blood cell). The Fr. 4 with the highest immunostimulating activity was subjected to DEAE-cellulose ion exchange chromatography and gave five fractions, 4-I, II, III, IV, and V. The in vitro immunostimulating assay of the five fractions showed that 4-I and 4-III had a similar activity to that of LPS but the other fractions had low activity. By analyses of chemical composition and HPLC, all fractions obtained were found to be heteropolysaccharide-protein complex. The molecular weights ranged from 9, 000 to 15, 000. Sugar analyses showed that glucose, galactose, mannose, arabinose, and xylose were main component. Uronic acid and amino sugar were also detected in the fractions. It should be noted that the molecular weight (15, 000) of 4-III was very small and the structure of 4-III may be different from the known immunostimulating branched $\beta$-(1longrightarrow3)-glucan.
Nitric Oxide (NO), derived from L-arginine, is produced by two types (constitutive and inducible) of nitric oxide synthase (NOS: cNOS and iNOS). The NO produced in large amounts by the iNOS is known to be responsible for the vasodilation and hypotension observed in septic shock, cancer metastasis and inflammation. The inhibitors of iNOS, thus, may be useful candidates for the treatment of inflammatory diseases accompanied by the overproduction of NO. We prepared alcoholic extracts of herbal drugs which have been used for the treatment of inflammation in oriental medicine. We have screened the inhibitory activity of NO production in lipopolysaccharide (LPS)-activated macrophages after the treatment of these extracts. Among 82 kinds of extracts of herbal drugs, 35 extracts showed the potent inhibitory activity of NO production above 50% at the concentration of $50\;{\mu}g/mL$. The inhibitory activities of NO production were also evaluated for several solvent fractions at two different concentrations. Especially, hexane and EtOAc fractions of Alpinia officinarum, Angelica gigas, Ostericum koreanum, Saussurea lappa, Torilis japonica, and hexane fractions of Agrimonia pilosa, Machilus thunbergii, Hydrangea serrata, Magnolia obovata, Prunella vulgaris, Tussilago farfara, and EtOAC fractions of Perilla frutescence showed a significant activity at 10 and/or $25\;{\mu}g/mL$. In Western blot analysis, the hexane fractions ($5\;{\mu}g/mL$) of Magnolia obovata and Saussurea lappa, and EtOAc fractions ($20\;{\mu}g/mL$) of Hydrangea Serrata, Perilla frutescence and Torilis japonica inhibited the expression of iNOS protein in LPS-activated macrophages. These plants may be promising candidates for the study of the activity-guided purification of active compounds and might be useful for the treatment of inflammatory diseases and endotoxemia accompanying overproduction of NO.
The objective of the trial was to study the relationship between the methane ($CH_4$) production and the Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate fractions of feeds for cattle and the suitability of CNCPS carbohydrate fractions as the dietary variables in modeling the $CH_4$ production in rumen fermentation. Forty-five rations for cattle with the concentrate/roughage ratios of 10:90, 20:80, 30:70, 40:60, and 50:50 were formulated as feed samples. The Menke and Steingass's gas test was used for the measurement of $CH_4$ production. The feed samples were incubated for 48 h and the $CH_4$ production was analyzed using gas chromatography. Statistical analysis indicated that the $CH_4$ production (mL) was closely correlated with the CNCPS carbohydrate fractions (g), i.e. CA (sugars); $CB_1$ (starch and pectin); $CB_2$ (available cell wall) in a multiple linear pattern: $CH_4=(89.16{\pm}14.93)$$CA+(124.10{\pm}13.90)$$CB_1+(30.58{\pm}11.72)$$CB_2+(3.28{\pm}7.19)$, $R^2=0.81$, p<0.0001, n = 45. Validation of the model using 10 rations indicated that the $CH_4$ production of the rations for cattle could accurately be predicted based on the CNCPS carbohydrate fractions. The trial indicated that the CNCPS carbohydrate fractions CA, $CB_1$ and $CB_2$ were suitable dietary variables for predicting the $CH_4$ production in rumen fermentation in vitro.
The forage potential of mulberry (Morus alba) was evaluated under Malaysian conditions. The yield and nutrient composition of plant fractions of mulberry (whole plant, leaf and stem fractions) were determined at four harvest stages, namely, 3 (W3), 5 (W5), 7 (W7) and 9 (W9) weeks in a randomized block design. The study was conducted over a 9-month period to assess the persistence of the crop to repeated harvests. Fresh and dry matter (DM) yields of all plant fractions increased significantly (p<0.01) with increasing intervals between harvests, with highest DM yields at W9. The leaf to stem ratio declined significantly (p<0.01) from 5.2 (W3) to 0.9 (W9), indicating predominance of the stem fraction with advancing maturity. The nutritional composition of plant fractions was also significantly influenced (p<0.01) by advancing plant maturity at harvest. Crude protein (CP), ash and the metabolisable energy content of plant fractions declined significantly (p<0.01) from W3 to W9, while there was a corresponding significant increase (p<0.01) in the acid detergent fibre, neutral detergent fibre and acid detergent lignin. From this study it was concluded that the optimum stage to harvest the whole plant is 5 weeks, which is a compromise between yield, nutrient composition (CP and fibre components), and the annual number of cuts, with good crop persistence to repeated harvests. Fresh mulberry whole plant can provide a valuable supplemental source of nutrients to poor quality basal diets.
The protein content of seed meal were examined to evaluate genotypes of higher protein content from 164 peanut germplasms. The variations in amino acids content were investigated from nine Korean leading varieties grown at five locations. Amino acids contents of protein fractions were also ana lysed in the present study, too. The results of the study were summarized as fallows. The seed protein of 164 peanut varieties averaged 24.5% ranging from 19.5% to 28.7%, showed 9.2% varietal variation in proteins. Differences were not observed in protein content between plant types, however, protein contents were higher in small-seeded than large seeded varieties. The differences were more greater particulaly in Spanish type varieties. The varieties introduced from Taiwan and Philippine showed higher protein contents, and the cultivars or lines bred in Korea and introduced from Japan were lower in protein contents. protein contents showed non-significantly negative correlations with 100-kernel weight and pod weight per plant, but positive correlations were observed between oil and protein content in all types of peanuts. Significant differences among the varieties and locations were observed for total, essential and non-essential amino acids contents, and aspartic acid. Locational differences for arginine, lysine, methionine, threonine, glutamic acid, glycine and tyrosine, and varietal difference for phenylalanene were revealed as significant. The limiting amino acids from the leading varieties were isoleucine, methionine, threonine, alanine and tyrosine, comparing with FAO recommanding levels of amino acids. Among the protein fractions prolamins was the highest in total amino acids, but essential amino acids was the highest in globulins
Previously, we discovered a new MMP-2 isoform GA110, of which appearance in human follicular fluid(FF) and serum was increased by EDTA. The present study was conducted to investigate how GAI 10 can appear by EDTA. To examine possible involvement of protein disulfide isomerase(PDI), an enzyme responsible for the dimerization of protein via disulfide formation, effect of PDI inhibitor on the appearance of GA110 by EDTA was investigated. When PDI inhibitor added to FF before EDTA treatment, the gelatinolytic activity of GA110 was abolished in a concentration dependent manner. By contrast, the activity of 72 kDa gelatinase increased. However, the PDI inhibitor added to FF after EDTA treatment, the gelatinolytic activity of GA110 was unaffected. To find out the nature of the enzyme which converts 72 kDa gelatinase into GAI 10, chromatographic separation method of FF proteins was done. Using hydroxyapatite column, fractions rich in 72 kDa gelatinase were isolated and pooled. By using this pool as substrate for the 72 kDa converting enzyme, protein fractions containing the converting activity were obtained from chromatographic separation of FF onto glutathione sepharose fast flow column. When immunoblotting was performed on this enzymatically active protein fractions against polyclonal anti-PDI antibody, distinct immunoreactivity was observed, although appeared in smaller molecular weight region. Based on these observations, it is suggested that the appearance of GAI 10 in FF by EDTA treatment could be due to an activation of PDI-like enzyme, which dimerizes 72 kDa gelatinase into GAI 10 via the formation of disulfide bond between molecules.
Prunus tomentosa Thunberg seed was investigated to evaluate its possibility for use as food resources of fats and proteins. The seed contained 40.38% of crude fat and 26.59% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.49% of neutral lipids, whereas compound lipids were only 4.51%. Among the neutral lipid components by thin-layer chromatography, triglycerides were 89.86%, sterols, monoglycerides, sterol esters, free fatty acids and diglycerides were 4.14%, 2.98%, 1.77%, 1.07%, and 0.18%, respectively. Oleic acid (65.06-66.05%) and linoleic acid (26.56-28.40%) were the main fatty acids in the total lipid, neutral lipid and triglyceride fractions. In the glycolipid and phospholipid fractions, predominant fatty acids were oleic acid (40.55-51.46%), linoleic acid (20.26-30.89%) and palmitic acid (17.64-21.43%). The extractability of salt soluble protein of seed was 60%, and recovery rate of main protein fraction separated by Sephadex G-200 was about 46.5%. The electrophoretic analysis showed 7 bands in seed protein.
Kim, Sung-Hwan;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull
Parasites, Hosts and Diseases
/
v.26
no.4
/
pp.239-244
/
1988
In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot
The contents of insoluble protein nitrogen, water soluble protein nitrogen anti peptides' nitrogen were determined of the samples which were taken in seven and half hours intervals during soybean-koji preparation to study changes of soybean protein, and the contents of total nitrogen and amino nitrogen were measured for the fractions resulting from molecular sieving by using Dowex 50 having various cross linkages for the peptides from soybean-koji extracts. As the results of the studies, The followings were obtained: 1. The contents of insoluble protein nitrogen and peptides' nitrogen are fairy constant at the earlier stage, where the former decreased and the latter increased markedly as mycelia grow, then rate of the decreases and the increases of them become lower at later stage after sporulation. The contents of water soluble protein are also constant at the earlier stage until covering of mycelia over the koji and increased since then until the stage of sporulation and then decreased at the later stage. 2. The amount of peptides nitrogen in each fraction obtained by the molecular sieving was almost constant at the earlier stage and the values in fractions of X-16, X-12, X-8. X-4 and X-2 increased considerably together as mycelia grow. Then the values in the fractions showed almost plateaux, after sporulalion, where the effluent fraction showed markedly increased values throughout mycelia growth.
Different types of bap(cooked rice) was cooked using barley or/and SoRiTae with rice as the base. Total(TS), rapidly digestible(RDS), slowly digestible(SDS) and resistant(RS) starch fractions were determined. Other physicochemical properties such as moisture, protein, amylose contents, protein digestion in vitro and color values as well as sensory properties of different bap were also investigated. Cooked rice with SoRiTae(RiSo) showed the highest moisture content of 63.9%, whereas other bap showed simillar content ranging from 62.3-63.0%. Crude protein content of RiSo was the highest, while that of cooked rice(Ri) was the lowest(p<0.05). Amylose content of RiBa was the highest, while that of RiSo was the lowest(p<0.05). In in vitro protein digestibility(IVPD), cooked rice with barley and SoRiTae(RiBaSo) was the highest, while Ri was the lowest, showing no significant difference at p<0.05. In starch fractions, as barley or/and SoRiTae were added to rice, a decrease in RDS content and increases in SDS and RS contents were observed. In addition, starch digestion index(SDI), which derived as an indicator of their in vitro starch digestibility and rapidly available glucose(RAG) value, which determined as a predictor of potential glycemic response decreased. A decrease in L value from RiSo and RiBaSo, which comprised of SoRiTae and increases in a and b values in RiSo and RiBa were observed, respectively. All sensory parameters involving color, glossiness, sweet taste, wetness, roughness, hardness and stickiness were shown to be a significant difference except sweet taste among different bap(p<0.05). L value of instrumental characteristic was negatively correlated with color of sensory characteristic and a value was positively correlated. Significant negative correlation was found between RS content and glossiness, however, positive correlation with roughness and hardness, respectively. These results suggested that cooked rice mixed with barley and SoRiTae contain significant RS and SDS contents and may improve diabetes and hyperlipidemia, due to the lowering RDS and RAG, respectively.
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