Basidiomycete PM2, a lignin-degrading white rot fungus, produces lgnin peroxidase (Lip) and manganese peroxidase (Mnp) in nutrient nitrogen limited liquid cultures. This fungus was selected for its ability to decolorize azo group of dyes. In order to improve production of the peroxidases and rapid dye decolorizing activity by basidiomycete PM2, the addition of veratryl alcohol or Tween 80 to nutrient nitrogen limited liquid cultures were tested. It was found to have a large stimulatory effect on Mnp activities and decolorization rate of azo dyes. A maximum Mnp activities of 254.2 u/L with veratryl alcohol and 192.2 u/L with Tween 80 were achieved respectively. These values were about 3.4-fold and 2.5-fold higher than that obtained in the control cultures (without alcohol or Tween 80), whereas the levels of Lip activity detected were very low (about 12 u/L)in all the cultures. In further experiments using three kinds of azo dyes of congo red, orange G and orange IV, enzyme activities and dye decolorization were investigated in the above-mentioned cultures. The results showed that Mnp activities and decolorization were notably higher than those obtained in the control cultures in the presence of azo dyes. Cultures supplemented with Tween 80 were more adequate for dye decolorization. The rates of the decolorization with Tween 80 of congo red (95.4%), orange G (98.5%) and orange IV (54.4%) after 24 hours of dye incubation were higher than that supplemented with veratryl alcohol. According to the results, Mnp activities secreted by basidiomycete PM2 play an essential role in the process of dye decolorization. Tween 80 was the main factor affecting the decolorization. The analysis of structure of the three kinds of azo dyes indicats that the extent of decolorization is affected by the dye molecular structure. The types and quantity of the substituted groups on the aromatic ring of azo dyes have effect on the percentage of biological decolorization.
Azo Compounds ; metabolism ; Basidiomycota ; enzymology ; metabolism ; Benzyl Alcohols ; pharmacology ; Coloring Agents ; metabolism ; Oxygenases ; biosynthesis ; Peroxidases ; biosynthesis ; Polysorbates ; pharmacology

In order to study the effect of phytohormone on growth and isoflavones contents of Pueraria phaseoloides hairy roots, we cultured the hairy roots with different concentrations of 6-benzylaminopurine (6-BA) alone or in combination with α-naphthaleneacetic acid (NAA). Then we determined the effects of 6-BA alone or in combination with NAA on the growth and the contents of isoflavones compounds and levels of antioxidase activities of hairy roots by spectrophotometry. The results show that 6-BA inhibited the growth, and decreased biomass and total isoflavones compounds of P. phaseoloides hairy roots. Furthermore, the inhibition was increased with the concentrations of 6-BA. Compared with the controls, different concentrations of 6-BA in combination with NAA 2.0 mg/L could inhibit the growth of hairy roots and decrease the content of total isoflavone compounds, and also significantly enhanced the contents of soluble protein and levels of peroxidase (POD) activities, but decreased the activities of superoxide dismutase (SOD). DNA ladders detected by agarose gel electrophoresis can be observed after hairy roots of P. phaseoloides were cultured with 6-BA alone for 30 days, but can appear on the 20th day after culture with 6-BA in combination with NAA 2.0 mg/L. This result indicates that 6-BA or 6-BA in combination with NAA can both stimulate appearance of programmed cell death (PCD), and NAA may play a synergistic role on PCD.
Benzyl Compounds ; Isoflavones ; chemistry ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Roots ; chemistry ; drug effects ; growth & development ; Pueraria ; drug effects ; growth & development ; Purines

PURPOSE: Itopride hydrochloride (itopride) inhibits acetylcholinesterase (AChE) and antagonizes dopamine D(2) receptor, and has been used as a gastroprokinetic agent. However, its prokinetic effect on the small bowel or colon has not yet been thoroughly investigated. The aim of this study was to investigate the effects of itopride on motor functions of the ileum and colon in guinea pigs. MATERIALS AND METHODS: The distal ileum was excised and the activity of peristaltic contraction was determined by measuring the amplitude and propagation velocity of peristaltic contraction. The distal colon was removed and connected to the chamber containing Krebs-Henseleit solution (K-H solution). Artificial fecal matter was inserted into the oral side of the lumen, and moved toward the anal side by intraluminal perfusion via peristaltic pump. Colonic transit times were measured by the time required for the artificial feces to move a total length of 10cm with 2-cm intervals. RESULTS: In the ileum, itopride accelerated peristaltic velocity at higher dosage (10(-10)-10(-6)M) whereas neostigmine accelerated it only with a lower dosage (10(-10)-10(-9)M). Dopamine (10(-8)M) decelerated the velocity that was recovered by itopride infusion. Itopride and neostigmine significantly shortened colonic transit at a higher dosage (10(-10)-10(-6)M). Dopamine (10(-8)M) delayed colonic transit time that was also recovered after infusion of itopride. CONCLUSION: Itopride has prokinetic effects on both the ileum and colon, which are regulated through inhibitory effects on AChE and antagonistic effects on dopamine D(2) receptor.
Animals ; Benzamides/*pharmacology ; Benzyl Compounds/*pharmacology ; Cholinesterase Inhibitors/pharmacology ; Colon/*drug effects/physiology ; Dopamine/pharmacology ; Dose-Response Relationship, Drug ; Gastrointestinal Motility/*drug effects ; Guinea Pigs ; Ileum/*drug effects/physiology ; Neostigmine/pharmacology ; Receptors, Dopamine D1/antagonists & inhibitors/physiology

OBJECTIVETo determine optimum culture conditions for the seed embryo culture and rapid propagation of Dendrobium candidum.

METHODSeed embryos of D. candidum were incubated in the medium containing a combination of 6-benzylaminopurine (BA) and 1-naphthaleneacetic acid (NAA), potato extract, banana extract and activated carbon in order to induce seed embryo germination, protocorm differentiation, plantlet propagation and plantlet rooting.

RESULT AND CONCLUSIONThe maximum embryo germination percentage was obtained in the 1/2 MS media supplemented with 20% potato extract. The 1/2 MS medium supplemented with 1.0 mg x L(-1) BA and 0.1 mg x L(-1) NAA was very beneficial to the protocorm differentiation and propagation of D. candidum. The highest protocorm propagation index was obtained from the medium containing the activated carbon. The highest root numbers and length were observed in plants growing in 1/2 MS medium containing 0.5 mg x L(-1) NAA.

Benzyl Compounds ; Carbon ; pharmacology ; Culture Media ; Dendrobium ; growth & development ; Germination ; drug effects ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; growth & development ; Purines ; Seeds ; growth & development ; Tissue Culture Techniques ; methods

OBJECTIVETo determine the content of baicalin in Scutellaria baicalensis callus induced by different doncentrations of exogenous hormones.

METHODHPLC system was adopted to determine baicalin in S. baicalensis callus. Chromatographic conditions: ODS column was adopted, with methanol-water-phosphate (47: 53: 0.2) as the mobile phase. The flow velocity was 1 mL x min(-1), the detective wavelength was 280 nm, and the temperature of column was room temperature.

RESULTS. baicalensis callus induced by 6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) showed the highest baicalin content, up to 49.78 mg x g(-1).

CONCLUSIONThe experiment is such a simple, rapid and stable method for determining the baicalin content that it can be used for determining the baicalin content in S. baicalensis callus.

2,4-Dichlorophenoxyacetic Acid ; pharmacology ; Benzyl Compounds ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Flavonoids ; metabolism ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Purines ; Scutellaria baicalensis ; drug effects ; metabolism ; Tissue Culture Techniques ; methods

OBJECTIVETo explore the difference of the shapes of fruits and germination characters of seeds of Rheum palmatum, R. tanguticum and R. officinale.

METHODThe seeds of three Rheum species including wild and cultivation one were collected from different regions. Character of the fruits appearance were observed. The purification, weights per thousand seeds, content of moisture, seed vigor and different germination rates of Radix et Rhizoma Rhei were measured. The germination rates of the seeds of R. palmatum were determined under different temperature and hormone treatment .

RESULTIt was markedly variant in shapes of the fruits, rates of germination and all quality characters of the three category of Radix et Rhizoma Rhei. As reported, the size of the fruit and seed and weights per thousand seeds of R. officinale was the biggest. The seed vigor and germination rate of R. palmatum caltivated in Ming county of Gangsu were 95.7% and 94% respectively. The rate of seed vigor was closely correlated with the rate of germination for other species. Results indicated that temperature had some effects on the germination, and there were obvious difference in the active effect of gibberellin, kinetin and 6-benzyl aminopurine on the germination of the seed. Our experimental results showed that the kinetin concentration of 50 x 10(-6) g x L(-1) had the most active effect, and its rates of germination increase 18 percent than the control one.

CONCLUSIONSignificant differences exist in the shapes of fruits, rates of germination of the seeds of Rheum, which are affected by the factors of temperatures and hormones. The result of this study provides scientific evidences for identify, growth and cultivation of Rheum.

Benzyl Compounds ; Fruit ; anatomy & histology ; Germination ; drug effects ; Gibberellins ; pharmacology ; Kinetin ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; anatomy & histology ; physiology ; Purines ; Rheum ; anatomy & histology ; physiology ; Seeds ; physiology ; Species Specificity ; Temperature

The effects of exogenous phytohormones on hairy root growth and biosynthesis of anthraquinones in the hairy root cultures of Polygonum multiflorum Thunb. were studied. The results showed that the 2,4-D, NAA and 6-BA all have obvious effects on the growth of hairy root cultures and the biosynthesis of anthraquinones. The growth of hairy root and biosynthesis of anthraquinones were strongly restrained by 2,4-D. However, NAA and 6-BA of appropriate concentration were favourable to hairy root growth and anthraquinones production.
2,4-Dichlorophenoxyacetic Acid ; pharmacology ; Anthraquinones ; metabolism ; Benzyl Compounds ; Chromatography, High Pressure Liquid ; Kinetin ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; metabolism ; Polygonum ; drug effects ; metabolism ; Purines

Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology. In the present study, we investigated the melanogenic activity of six structurally distinct compounds, namely, scopoletin, kaempferol, chrysin, vitamin D, piperine, and 6-benzylaminopurine. We determined their effectiveness, toxicity, and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model. The melanogenic activity of 6-benzylaminopurine, the compound identified as the most potent, was further verified by measuring green fluorescent protein concentration in tyrp1 a: eGFP (tyrosinase-related protein 1) zebrafish and mitfa: eGFP (microphthalmia associated transcription factor) zebrafish and antioxidative activity. All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF. 6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20 μmol·Lin vivo and 100 μmol·Lin vitro, and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1a: eGFP and mitfa: eGFP zebrafish in vitro. However, its relative anti-oxidative activity was found to be very low. Overall, our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism, by increasing melanin content via positive regulation of tyrosinase activity in vitro, as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.
Alkaloids ; chemistry ; pharmacology ; Animals ; Benzodioxoles ; chemistry ; pharmacology ; Benzyl Compounds ; chemistry ; pharmacology ; Cholecalciferol ; chemistry ; pharmacology ; Flavonoids ; chemistry ; pharmacology ; Humans ; Kaempferols ; chemistry ; pharmacology ; Melanins ; genetics ; metabolism ; Monophenol Monooxygenase ; genetics ; metabolism ; Pigmentation ; drug effects ; Piperidines ; chemistry ; pharmacology ; Polyunsaturated Alkamides ; chemistry ; pharmacology ; Purines ; chemistry ; pharmacology ; Scopoletin ; chemistry ; pharmacology ; Vitiligo ; drug therapy ; enzymology ; metabolism ; Zebrafish

In a preliminary study, we found that benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD- fmk), unlike Boc-aspartyl(OMe)-fluoromethylketone (BocD-fmk), at usual dosage could not prevent genistein-induced apoptosis of p815 mastocytoma cells. This study was undertaken to reveal the mechanism underlying the incapability of zVAD-fmk in preventing this type of apoptosis. We observed that 14-3-3 protein level was reduced in genistein-treated cells and that BocD-fmk but not zVAD-fmk prevented the reduction of 14-3-3 protein level and the release of Bad from 14-3-3. We also demonstrated that truncated Bad to Bcl-xL interaction in genistein- treated cells was prevented by BocD-fmk but not by zVAD-fmk treatment. Our data indicate that BocD- fmk, compared to zVAD-fmk, has a certain preference for inhibiting 14-3-3/Bad signalling pathway. We also elucidated that this differential efficacy of BocD-fmk and zVAD-fmk resulted from the different effect in inhibiting caspase-6 and that co-treatment of zVAD-fmk and caspase-6 specific inhibitor substantially prevented genistein-induced apoptosis. Our data shows that caspase-6 plays a role on Bad/14-3-3 pathway in genistein-induced apoptosis of p815 cells, and that the usual dose of zVAD-fmk, in contrast to BocD-fmk, did not prevent caspase-6 acting on 14-3-3/Bad-mediated event.
bcl-Associated Death Protein/*metabolism ; *Signal Transduction/drug effects ; Mitochondria/drug effects ; Mice ; Mastocytoma ; Hydrocarbons, Fluorinated/*pharmacology ; Genistein/pharmacology ; Enzyme Inhibitors/*pharmacology ; Cell Line, Tumor ; Caspase 6/antagonists & inhibitors/*metabolism ; Benzyl Compounds/*pharmacology ; Apoptosis/*drug effects ; Animals ; Amino Acid Chloromethyl Ketones/pharmacology ; 14-3-3 Proteins/*metabolism

OBJECTIVETo study the effect of sucrose and plant growth substances of different concentrations on the induction of test-tube tuberous roots of Rehmannia glutinosa, in order to establish an efficient system for the induction of test-tube tuberous roots from leaves of R. glutinosa.

METHODLeaves from test-tube seedlings of 85-5 R. glutinosa were used as explants. After rooting induction, they were transferred to medium with orthogonal design for inducing test-tube tuberous roots of R. glutinosa.

RESULT AND CONCLUSIONNAA played a significant role in induction of test-tube tuberous roots of R. glutinosa, followed by sucrose and 6-BA. With leaves from test-tube seedlings as the explants, the optimal medium for inducing test-tube tuberous roots of R. glutinosa was MS + BA 3.0 mg x L(-1) + NAA 0.1 mg x L(-1) + sucrose 7%. The study provides an efficient induction system for studies on artificial seeds and secondary metabolism with test-tube tuberous roots of R. glutinosa.

Benzyl Compounds ; Dose-Response Relationship, Drug ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; drug effects ; growth & development ; Plant Roots ; drug effects ; growth & development ; Purines ; Rehmannia ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Sucrose ; pharmacology ; Tissue Culture Techniques ; instrumentation ; methods