April - June 2007 - Kasetsart University

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316 compound exerted weak anti HSV-1 activity when compared with that of the crude hot water polysaccharide, whereas, in the absence of sulfate groups in polysaccharide, no significant anti HSV-1 activity was detected in this compound (Table 5). DISCUSSION This study found that the hot water extract polysaccharide exhibited anti HSV-1 activity, while the cold extract of the polysaccharide did not. Previous studies found that the majority of potential antiviral algal polysaccharides were extracted from tissues by hot water, dilute acid or alkali solution (Damonte et al., 1994; Hoshino et al., 1998). Crude hot water polysaccharide still contained a high level of protein which may co-precipitate when CTAB is <strong>Kasetsart</strong> J. (Nat. Sci.) 41(2) used for polysaccharide precipitation (Tomanee et al., 2004). Partial purification of the hot water polysaccharide by gel-filtration on Sepharose 6B column gave 2 fractions (SHP-F1 and SHP-F2), both fractions effectively inhibited HSV-1 activity. Results reported by Hayashi et al. (1996a) revealed 3 fractions (SP-H-1, SP-H-2 and SP-H-3) but only a SP-H-2 fraction had anti HSV-1 activity. The sugars found in SHP-F1 and SHP-F2 fractions in this study are almost the same as previously reported by Hayashi et al. (1966a) except for arabinose which was found in this study instead of fructose which was reported by the same researchers. Calcium ion and sulfate groups in the hot water polysaccharide were important for the anti HSV-1 activity. This result was supported by Hayashi’s study that when the calcium-free Table 4 Carbohydrate and protein content of crude hot water polysaccharide which was precipitated by trichloroacetic acid (TCA). TCA concentration (%) % w/w of crude hot water polysaccharide Carbohydrate Protein 0 42.5 ± 0.3 31.0 ± 0.8 10 41.8 ± 1.5 28.1 ± 2.1 20 45.3 ± 1.8 25.4 ± 1.3 30 49.2 ± 2.3 22.3 ± 1.5 50 53.1 ± 2.0 18.0 ± 0.8 Mean ± standard deviation (n = 3) Table 5 Anti HSV-1 activity in the crude hot water polysaccharides from S. platensis. Sample Cytotoxicity a (IC 50 : µg/ml) Anti HSV-1 b (IC 50 : µg/ml) Polysaccharide > 50 21.3 Polysaccharide (-Ca 2+ ) > 50 38.4 Polysaccharide (-SO 4 2- ) > 50 Inactive a maximum concentration of compound for cytotoxicity test was 50 µg/ml compound was non toxic on the growth of Vero cells if IC 50 >50 µg/ml (if compound was toxic on the growth of Vero cells, the compound will be subjected to the serial dilution for determination of IC 50 value) b % inhibition of HSV-1; < 25%= inactive, 25-35%= weakly active, >35-50% = moderately active, > 50%= active (the compound will be subjected to the serial dilution for determination of IC 50 value)
spirulan (H-SP), and a desulfated compound from Ca-SP were subjected to cytotoxicity and antiviral assay, both compounds exerted strong toxicity to the growth of host cell (HeLa cells) and weakly inhibited HSV-1 (Hayashi et al., 1996a). Ca-SP was found to inhibit replication of several enveloped virus and selectively inhibited the penetration of virus into host cell (Hayashi et al., 1996b). Loya et al. (1998) postulated that the negatively charged (e.g., sulfonate vs. sulfate) may interact with the positively charged side chains on the DNA polymerase and Witvrouw et al. (1994) assumed that sulfated polysaccharides disruption of ionic interactions between positively charged regions of viral surface glycoproteins and cellular membrane phospholipids. CONCLUSION Results from this study demonstrated the significant potential of S. platensis polysaccharide for activity against HSV-1. The hot water extract polysaccharide which contained rhamnose as the main sugar component showed anti HSV-1 activity at IC 50 21.3 µg/ml. Calcium ion and sulfate groups in the polysaccharide had major roles in the anti HSV-1 activity. S. platensis, is a possible source for new drugs in the treatment of HSV-1 and other viral diseases. ACKNOWLEDGEMENTS This study was supported by TRF (The Thailand Research Fund, RDG4330032). LITERATURE CITED Blakeney, A. B., P. J. Harris, R. J. Henry and B. A. Stone. 1983. A simple and rapid preparation of alditol acetates for monosaccharide analysis. Carbohydr. Res. 113: 291-299. Burns, B. A. 1995. In Official Methods of Analysis of Association of Official Analytical Chemists <strong>Kasetsart</strong> J. (Nat. Sci.) 41(2) 317 International, Chapter 5, 16 th Ed Volume II, Edited by Cunniff, P., AOAC International, Virginia, p. 5 Damonte, E. B., J. Neyts, C. A. Pujol, R. Snoeck, G. Andrei, S. Ikeda, M. Witvrouw, D. Reymen, H. Haines and M. C. Matulewicz. 1994. Antiviral activity of a sulfated polysaccharide from the red seaweed Nothogenia fastigiata. Biochem Pharmacol. 47: 2187-2192. Dubois, M., K. A. Gilles, J. K. Hamilton, P. A. Rebers and F. Smith. 1956. Colorimetric Method for Determination of Sugars and Related Substances. Anal. Chem. 28: 350- 356. Elion, G. B., P. A. Furman, J. A. Fyfe, P. De Miranda, L. Beauchamp and H. J. Schaeffer. 1977. Selectivity of action of an antiherpetic agent, 9-guanine. Proc. Natl. Acad. Sci. USA. 74: 5716-5720. Field, A. K. and K. K. Biron. 1994. The end of innocence revisited: resistance of herpes viruses to antiviral drugs. Clin. Microbiol. Rev. 7: 1-13. Folch, J., M. Lees and G. H. S. Stanley. 1957. A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem. 226: 497-509. Gerber, P., J. D. Dutcher, E. V. Adams and J. H. Sherman. 1958. Protective effect of seaweed extracts for chicken embryos infected with influenza B or mumps virus. Proc. Soc. Exp. Biol. Med. 99: 590-593. Hayakawa, Y., T. Hayashi, K. Hayashi, K. Ozawa, K. Niiya and N. Sakuragawa. 1996. Heparin cofactor II-dependent antithrombin activity of calcium spirulan. Blood Coagul. Fibrinol. 7: 554-560. Hayashi, K., T. Hayashi and M. Morita. 1993. An extract from Spirulina platensis is a selective inhibitor of Herpes simplex virus type 1 penetration into HeLa cells. Phytother. Res. 7: 76-80. Hayashi, T., K. Hayashi, M. Maeda and I. Kojima.
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April - June 2007 Volume 41 Number
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KASETSART JOURNAL NATURAL SCIENCE T
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Kasetsart J. (Nat. Sci.) 41 : 205 -
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harvesting time which started from
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y Chen and Paull (2000). Figure 1 a
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pineapple fruit allowed the accumul
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Kasetsart J. (Nat. Sci.) 41(2) 215
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Cluster analysis Cluster analysis u
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Table 3 (Continued) 1 2 3 4 5 6 7 8
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CONCLUSION AFLP markers classified
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difference of soil texture used in
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under wet soil condition planting.
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tropics. Following the expansion, w
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sealed in a plastic box with 1 cm w
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moisture content increment after so
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Frequency Frequency 30 20 10 0 20.0
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School, Kasetsart University. The a
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for combining ability of lines (Lam
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selective mass sibbing within indiv
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Although most of S 2-interfamily hy
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composite sets as used in this stud
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days. The numbers of calli producin
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the report of Ching (1982) showing
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clearly amplified bands generated b
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caused by either the recombination
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Kuhlmann, V. and B. Foroughi-Wehr.
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Xanthomonas fuscans subsp. aurantif
Page 65 and 66: Kasetsart J. (Nat. Sci.) 41(2) 265
Page 67 and 68: was collected and washed with 70% e
Page 69 and 70: expected size was amplified with 35
Page 71 and 72: eaction technique has been used for
Page 73 and 74: Schaad, N.W., D. Opgenorth and P. G
Page 75 and 76: MATERIALS AND METHODS Model descrip
Page 79 and 80: calculated TF value calculated TF v
Page 81 and 82: sincere gratitude and deep apprecia
Page 83 and 84: 1989). In monogastrics, the inclusi
Page 85 and 86: of all LLS samples in this study we
Page 87 and 88: Melbourne, Parkville, Victoria. Goe
Page 89 and 90: considered responsible for low prod
Page 93 and 94: mineral supplementation suggested b
Page 95 and 96: Body weight chane (kg/head) 32 30 2
Page 97 and 98: CONCLUSION AND RECOMMENDATION With
Page 99 and 100: SAS. (Statistical Analysis System).
Page 101 and 102: genes covering a variety of desirab
Page 103 and 104: mg/l NAA and 0.5 mg/l zeatin) incub
Page 105 and 106: 5. Purification by various sucrose
Page 107 and 108: as the culture period increased. So
Page 109 and 110: culture, pp. 1-20. In L.C. Fowke an
Page 111 and 112: Kasetsart J. (Nat. Sci.) 41 : 311 -
Page 113 and 114: GC. Analytical methods Total carboh
Page 115: ash. The crude hot water polysaccha
Page 121 and 122: cells often displayed an increased
Page 123 and 124: LITERATURE CITED Bell, T.A. and D.V
Page 125 and 126: for alternative sources of supply.
Page 127 and 128: BR2.1.2 formed the same clade with
Page 129 and 130: supplement with glutamate (Iida et
Page 131 and 132: optimal for S. limacinum BR2.1.2 fo
Page 133 and 134: fluorescent lamp at 1.5 kLux develo
Page 137 and 138: Wisconsin, USA). Total extracted RN
Page 139 and 140: RESULTS Cloning and sequencing of m
Page 141 and 142: Expression of rmIL-2 and purificati
Page 143 and 144: other strains of mice have differen
Page 145 and 146: dose interleukin-2 therapy promotes
Page 147 and 148: The chitosano-oligosaccharides are
Page 149 and 150: enzyme and cells were maximized by
Page 151 and 152: of crude chitosanases. Fortunately,
Page 153 and 154: Relative activity (%) 100 80 60 40
Page 155 and 156: Uchida, K. Tateishi, O. Shida and K
Page 157 and 158: MATERIALS AND METHODS 1. Materials
Page 159 and 160: without coating and coated with pec
Page 161 and 162: in Table 5-7. It was found that the
Page 165 and 166: the cultures at 10,200 × g for 15
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incubation and reached the final pH
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y lysine- and tyrosine-decarboxylas
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during the stages of ripening as su
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As the 25 companies were divided in
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All the data comes up with informat
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as piece “A” or straight as pie
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et al. (1998); Zeng (2000); and Tal
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p 1, i * Kasetsart J. (Nat. Sci.) 4
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c ≤ c2 jq2 j n q p k p * 2, j , ,
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algorithm would consider RM 5, prio
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the maximum deviation of about 10%.
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Stock Quantities Using Heuristic Op
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which dynamically and automatically
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2. Framework architecture and compo
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Risk (VaR) calculation which was im
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Throughput (job/sec) 7 6 5 4 3 2 1
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Speed up 40 35 30 25 20 15 10 5 0 F
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and discovery, resource selection a
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April - June 2007 - Kasetsart University

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