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Two in vivo experiments were conducted to evaluate the potential of Pharbitis nil seeds (PA) as an anti-methanogenic additive to ruminant feed. In experiment 1, six Hanwoo steers (459.0 ± 25.8 kg) were fed either a total mixed ration (TMR; 32-d period) or TMR supplemented with PA at 5% dry matter (DM) intake (TMR-PA; 45-d period) for two consecutive periods. Fecal and urine outputs were measured in an apparent digestibility trial in both periods. Methane (CH4) yield and heat energy (HE) were measured using respiratory chambers equipped with gas analyzers. In experiment 2, five rumen cannulated Holstein steers (744 ± 35 kg) were fed the same TMR or TMR-PA diets for 40 days; rumen samples were collected at 0, 1.5, and 3 h after feeding on the last day of the feeding period. In experiment 1, although there were no differences (p > 0.05) in nutrients or gross energy intake (GEI) between the groups, an increase (p < 0.05) in the apparent digestibility of DM (9.1%) and neutral detergent fiber (22.9%) was observed in the TMR-PA fed Hanwoo steers. Pronounced decreases (p < 0.05) in CH4 (g/Kg DM; 17.1%) and urinary N excretion (% N intake; 7.6%) were observed in the TMR-PA group, leading to a 14.7% increase in metabolizable energy intake (% GEI). However, only a numerical increase (p > 0.05) in retained energy was observed due to the increase in HE loss. In experiment 2, a drastic decrease (p < 0.05) in rumen ammonia concentration (56.3%) associated with an increased (p = 0.091) rumen short-chain fatty acid concentration 1.5 h after feeding were observed in TMR-PA fed Holstein steers. A 26.6% increase (p < 0.05) in the propionate proportion during the treatment period clearly reflected a shift in the ruminal H2 sink after 3 h of feeding. A 40% reduction (p = 0.067) in the relative abundance of rumen protozoa Entodinium caudatum was also observed. It was concluded that PA could be a natural feed additive for CH4 and N emission abatement.

Abstract Background Pharbitidis Semen (the seeds of Pharbitis nil), traditionally used as a purgative in Japan, China and Korea, contains a resin glycoside fraction named pharbitin, which is known as a purgative ingredient. Due to the complex nature of pharbitin, little is known about either the action on intestinal tension caused by resin glycoside itself or by its components. Methods In this study, we investigated the effects of pharbitin, the glycosidic acid fraction (pharbitic acid) and the aglycone fraction (phar-genin) generated from pharbitin on peristalsis of colon and ileum isolated from mice with the Magnus method. Results We demonstrated that pharbitin (3–30 μg/mL) concentration-dependently increased tonus of mice colon via acetylcholine receptors, its components phar-genin (1.27–12.7 μg/mL) and pharbitic acid (10–1000 μg/mL) also had the increment on colon tonus. On the other hand, ileum tension decreased in the presence of pharbitin. Conclusions The effects of resin glycoside of Pharbitidis Semen on colon tonus are different with those on ileum tonus isolated from mice. In the next step it is necessary to investigate details of its pharmacological mechanism.

Two in vivo experiments were conducted to evaluate the potential of Pharbitis nil seeds (PA) as an anti-methanogenic additive to ruminant feed. In experiment 1, six Hanwoo steers (459.0 ± 25.8 kg) were fed either a total mixed ration (TMR; 32-d period) or TMR supplemented with PA at 5% dry matter (DM) intake (TMR-PA; 45-d period) for two consecutive periods. Fecal and urine outputs were measured in an apparent digestibility trial in both periods. Methane (CH 4 ) yield and heat energy (HE) were measured using respiratory chambers equipped with gas analyzers. In experiment 2, five rumen cannulated Holstein steers (744 ± 35 kg) were fed the same TMR or TMR-PA diets for 40 days; rumen samples were collected at 0, 1.5, and 3 h after feeding on the last day of the feeding period. In experiment 1, although there were no differences ( p > 0.05) in nutrients or gross energy intake (GEI) between the groups, an increase ( p < 0.05) in the apparent digestibility of DM (9.1%) and neutral detergent fiber (22.9%) was observed in the TMR-PA fed Hanwoo steers. Pronounced decreases ( p < 0.05) in CH 4 (g/Kg DM; 17.1%) and urinary N excretion (% N intake; 7.6%) were observed in the TMR-PA group, leading to a 14.7% increase in metabolizable energy intake (% GEI). However, only a numerical increase ( p > 0.05) in retained energy was observed due to the increase in HE loss. In experiment 2, a drastic decrease ( p < 0.05) in rumen ammonia concentration (56.3%) associated with an increased ( p = 0.091) rumen short-chain fatty acid concentration 1.5 h after feeding were observed in TMR-PA fed Holstein steers. A 26.6% increase ( p < 0.05) in the propionate proportion during the treatment period clearly reflected a shift in the ruminal H 2 sink after 3 h of feeding. A 40% reduction ( p = 0.067) in the relative abundance of rumen protozoa Entodinium caudatum was also observed. It was concluded that PA could be a natural feed additive for CH 4 and N emission abatement., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Bharanidharan, Xaysana, Hong, Kim, Byun, Lee, Tomple, Kim and Ibidhi.)

Background: Pharbitidis Semen (the seeds of Pharbitis nil), traditionally used as a purgative in Japan, China and Korea, contains a resin glycoside fraction named pharbitin, which is known as a purgative ingredient. Due to the complex nature of pharbitin, little is known about either the action on intestinal tension caused by resin glycoside itself or by its components. Methods: In this study, we investigated the effects of pharbitin, the glycosidic acid fraction (pharbitic acid) and the aglycone fraction (phar-genin) generated from pharbitin on peristalsis of colon and ileum isolated from mice with the Magnus method. Results: We demonstrated that pharbitin (3–30 μg/mL) concentration-dependently increased tonus of mice colon via acetylcholine receptors, its components phar-genin (1.27–12.7 μg/mL) and pharbitic acid (10–1000 μg/mL) also had the increment on colon tonus. On the other hand, ileum tension decreased in the presence of pharbitin. Conclusions: The effects of resin glycoside of Pharbitidis Semen on colon tonus are different with those on ileum tonus isolated from mice. In the next step it is necessary to investigate details of its pharmacological mechanism. [ABSTRACT FROM AUTHOR]

Context: Antiacetylcholinesterase (AChE) drugs have been a main therapeutic treatment for Alzheimer’s disease because increased AChE levels play a key role in reducing neurotransmission. Objectives: Extracts from 35 Korean plants were selected and screened for antioxidant and anti-cholinesterase activity to explore new sources derived from Korean natural resources that could be used as AD therapeutic agents. Materials and methods: The antioxidant effect of extracts from 35 selected Korean plants was determined using two most common free radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS). Additionally, the effect of extracts, identified as antioxidants, on acetylcholinesterase inhibition was assessed by an acetylcholinesterase assay kit. Results: Out of 36 extracts of 35 plants tested, Oenothera biennis L. (9.09 μg/mL), Saururus chinensis (Lour.) Baill. (9.52 μg/mL) and Betula platyphylla var. japonica (9.85 μg/mL) showed strong DPPH scavenging activity. Twelve other extracts also exerted moderate free radical scavenging activities with IC50 values ranging from 10 to 50 μg/mL. Antioxidant capacity detected by ABTS assay was only significant in O. biennis (23.40 μg/mL), while the other extracts were weak or unable to reduce the production of ABTS. Based on the antioxidant activities of these plant extracts, 19 extracts with IC50 values less than 100 μg/mL in DPPH assay were selected for further AChE inhibition assay. Among the extracts tested, the IC50 value for Prunella vulgaris var. lilacina NAKAI (18.83 μg/mL) in AChE inhibitory activity was the lowest, followed by O. biennis (20.09 μg/mL) and Pharbitis nil Chosy (22.79 μg/mL). Conclusions: Considering complex multifactorial etiology of AD, the extracts of P. vulgaris var. lilacina (aerial part), O. biennis (seed) and P. nil (seed) may be safe and ideal candidates for future AD modifying therapies.

Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven colorectal cancer (CRC) is notorious to target with drugs and has shown ineffective treatment response. The seeds of Pharbitis nil, also known as morning glory, have been used as traditional medicine in East Asia. We focused on whether Pharbitis nil seeds have a suppressive effect on mutated KRAS-driven CRC as well as reserving muscle cell functions during CRC progression. Seeds of Pharbitis nil (Pharbitis semen) were separated by chromatography and the active compound of Pharbitis semen (PN) was purified by HPLC. The compound PN efficiently suppressed the proliferation of mutated KRAS-driven CRC cells and their clonogenic potentials in a concentration-dependent manner. It also induced apoptosis of SW480 human colon cancer cells and cell cycle arrest at the G2/M phase. The CRC related pathways, including RAS/ERK and AKT/mTOR, were assessed and PN reduced the phosphorylation of AKT and mTOR. Furthermore, PN preserved muscle cell proliferation and myotube formation in cancer conditioned media. In summary, PN significantly suppressed mutated KRAS-driven cell growth and reserved muscle cell function. Based on the current study, PN could be considered as a promising starting point for the development of a nature-derived drug against KRAS-mutated CRC progression.

Exogenous acetylcholine (ACh), acetylcholinesterase (AChE) inhibitors, as well as agonists and antagonists of ACh receptors applied on the cotyledons of 5-day-old seedlings of Pharbitis nil during a 16-hour long inductive night or during a 12-hour long subinductive night, do not essentially influence the flower bud formation. Also the application of above mentioned substances to the seedlings growing under the conditions of 72 hours of darkness, 24 hours of light and then 24 hours of darkness does not influence in an essential way flowering of P. nil. On the other hand, applying these substances on the cotyledons of P. nil during 24-hour-long inductive night, preceded by 72 hours of darkness and then 24 hours of light of lowered intensity finished by 15-minute-long impulse of far red light which inhibit flowering, caused the reversion of the effect of far red light irradiation and stimulated the flowering. The obtained results suggest that endogenous system ACh/AChE could participate in the mechanism of a phytochrome controlled flowering of short day plants.

The endogenous content of ABA in the cotyledons of Pharbitis nil is high during the light phase before an inductive 16-h-long dark period. During the night, however, at the beginning, the level of ABA is relatively low with the tendency to increase during the second half of an inductive dark period. The dual effect of exogenous ABA on the Pharbitis nil flowering has been observed. ABA applied to the cotyledons on subthreshold photoperiod (12-h-long night) stimulates flower bud formation. On the other hand, however, ABA applied during an inductive (16-h-long) dark period, as well as applied to the medium of cultured plantlets, inhibits flowering. Thus, the flowering effect of ABA is clearly dependent on the state of flower induction which is different in plants growing on various photoperiods.

The aim of this work was to determine if there is any relationship between an endogenous phyA level and photoperiodic flower induction. The level of phyA was characterised with polyclonal antibodies directed to phyA from pea. At first it was detected that phyA level is predominant in cotyledons, whereas in roots and stems the concentration of labile phytochrome is rather low. So cotyledons were used for later experiments. In these cotyledons exposed to light illumination a rapid destruction of phyA has been observed. The loss of extractable phyA chromoprotein occurs already after 60 min of irradiation. Pharbitis nil is a short-day plant and a single 16-hours-long dark period is fully inductive. We assessed that phyA level is extremely low during a long inductive night and an immunodetectable phytochrome appears only after 24 hours of darkness. The obtained results suggest that labile phytochrome is not taking part in the direct control of the photoperiodic flower induction.

Seedlings of Pharbitis nil cultivated under non-inductive conditions of white light were subjected to generative induction applying one 16-hour-long period of inductive night. During the eighth hour the night was interrupted with 1 min of red light pulse which completely inhibited the flowering. Treating the plants with KCN blocked the inhibiting effect of red light. Because KCN lowers considerably the rate of destruction of labile Pfd in some plant systems, it seems probable that red light night-break irradiation (without KCN), which blocked the flowering, leads also to the accumulation of unknown Pfd destruction products (irreversible phytochrome). It also suggests that it is not the labile PfrI itself but the products of its irreversible transformation that could be active in the photoperiodic control of flowering.

The content of endogenous ethylene in the seedlings of Pharbitis nil subjected to 16-hour long inductive night is low during the first half of a dark period, then it increases considerably in the second half of the night. Ethrel, the compound releasing ethylene, applied to the cotyledons of the seedlings, increases the amount of endogenous ethylene in them and at the same time inhibits the flowering, especially when ethrel was applied during the first half of an inductive night, when the content of endogenous ethylene in the seedlings is low. The auxin, inhibiting the flowering of Pharbitis, causes at the same time the increase in the production of endogenous ethylene. PCIB, an inhibitor of auxin action reverses the inhibiting influence of ethrel on flowering. On the other hand the combined application of ethrel and TIBA, the inhibitor of auxin polar transport, causes the increase of the flowering inhibition. CoCl2, the inhibitor of ethylene biosynthesis, and AgNO3, the inhibitor of ethylene action, reverse partly the inhibiting influence of auxin. It suggests that ethylene could take part in auxininhibition of flowering. The all obtained results seem to suggest the participation of ethylene in the control of the flower photoperiodic induction.

The endogenous content of IAA in the cotyledons of Pharbitis nil is low before and during the first half of the inductive 16-h-long dark period. From the 8th to the 12th hour the level of IAA increased and then again was going down at the end of a dark period. Exogenous IAA applied to the cotyledones before and during the first half of the inductive dark period inhibits flower bud formation. The application of IAA to shoot apex also resulted in the inhibition of flowering. Experiments with TIBA, an auxin polar transport inhibitor, and PCIB, an auxin action inhibitor, have shown that auxin polar transport in cotyledones and long-distance auxin transport from cotyledones to shoot apex play an important role in IAA inhibition of flower bud formation. It suggests that auxins play their role not only at the level of floral induction in cotyledones, but also in the later events of floral evocation and differentiation in shoot apex.

The short-day plant, Pharbitis nil, requires only a single inductive cycle with a 16-hour dark period for flowering. The mitotic activity in the shoot apices was studied directly after the termination of the inductive photoperiod. A pronounced rise in the mitotic index was found in the 2nd and the 8th-14th hours. Control plants grown under noninductive conditions (continuous light, a light interruption in the middle of the dark period) did not flower and did not show an increased mitotic index. The increased mitotic activity in the shoot apices of Pharbitis seems to be causally connected with the phytochrome-controlled entry of the plants into the state of generative induction.

Ethnopharmacological relevance Pharbitis Nil (PN) is used as a main component of the existing drug, DA-9701, which was developed to treat functional dyspepsia (FD) in Korea. PN extracts isolated from its seeds have been reported to have anticancer effects. Aim of the study The purpose of this study was to investigate the underlying mechanism of the chemotherapeutic effects of PN in lung cancer cells. Materials and methods We performed MTT assays, colony formation assays, flow cytometry assays, Western blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence analysis, and cell counting assays to study the molecular mechanism of chemotherapeutic effects of PN in lung cancer cells. Results Our results indicate that PN induced autophagy as well as apoptosis. PN inhibited cell proliferation and survival by inducing apoptosis in several lung cancer cell lines. PN-treated cells also exhibited induction of autophagy, as evidenced by increased protein expression levels and punctuate patterns of LC3 II. Moreover, activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), which plays an important role in autophagy activation, was shown to be related with PN-induced autophagy. Interestingly, pharmacological blockade of autophagy activation with wortmannin and inhibition of ERK1/2 phosphorylation by U0126 markedly enhanced PN-induced apoptosis and reduced cell viability, suggesting that autophagy induced by PN may have a cytoprotective effect by suppressing apoptosis. PN- induced apoptosis was regulated by signal transducer and activator of transcription 3 (STAT3) deactivation. Moreover, decrease of STAT3 activation in PN-treated cells was associated with reduced survivin expression, further demonstrating that PN-induced apoptosis was regulated by STAT3 deactivation. Conclusion We believe that PN, which is already proven to treat human patients with FD, might be a potential anticancer drug for human lung cancer. In addition, our data suggest that the combination of PN treatment with an autophagy inhibitor or traditional anticancer agents may be an effective anticancer therapy. [ABSTRACT FROM AUTHOR]

Phytochemical investigation of an EtOH extract of Pharbitis nil seeds (Convolvulaceae) resulted in the isolation and identification of a new neolignan, 7 R ,8S- threo -dihydroxydehydrodiconiferyl alcohol ( 1 ), and a new monoterpene glycoside, (3 Z ,7 S )-7-hydroxy-3,7-dimethyl-3,8-octadienyl-β- d -glucopyranoside ( 2 ), together with a known compound, ethyl α- l -arabinofuranoside ( 3 ). The chemical structures of these compounds were unambiguously determined using physical data, HR-ESI–MS and spectroscopic evidence, including 1D and 2D NMR experiments. The anti-inflammatory activities of the isolates were evaluated by estimating the inhibition of nitric oxide (NO) production. Compounds 1 and 2 reduced NO levels in lipopolysaccharide (LPS)-stimulated murine microglial BV-2 cells. In addition, compound 2 showed weak cytotoxicity against the HCT-15 cell line with an IC 50 value of 28.6 μM. [ABSTRACT FROM AUTHOR]

Application of NH 2 silica gel column chromatography using CH 2 Cl 2 -MeOH was found to show a satisfactory resolution for separation of the crude resin glycosides of Pharbitis Semen (the seeds of Pharbitis nil ), led to the isolation of seven new acylated glycosidic acid methyl esters, Pharesinosides A-G ( 1 – 7 ), along with four known ones ( 8 – 11 ). These compounds ( 1 – 11 ) were considered to be generated via methyl esterification of the carboxyl group in acylated glycosidic acids. Their structures including stereochemistry were elucidated on the basis of a combination of the spectroscopic and chemical methods. All isolates were evaluated for anti-tumor migration activities with human colon cancer cell line HCT-116, and compound 7 exhibited a potent migration inhibitory activity. [ABSTRACT FROM AUTHOR]

A new ent-kaurane diterpene glycoside, pharboside H ( 1), and three known compounds, benzyl β-D-glucopyranoside ( 2), 1-(4′-O-glucopyranosyl-4′-hydroxy-3′,5′-dimethoxyphenyl)-2{2′′,6′′-dimethoxy-4′′-[1-(E) propen-3-ol]-phenoxy}-propane-l,3-diol ( 3), and butyl caffeate ( 4), were isolated from the seeds of Pharbitis nil. The structure of the new compound 1 was elucidated by 1D and 2D data analysis and enzyme hydrolysis. All the isolated compounds 1-4 were reported from this source for the first time. Compounds 1-4 were tested for cytotoxicity against four human tumor cell lines in vitro using the sulforhodamine B bioassay. [ABSTRACT FROM AUTHOR]

The estimation of participation and density of forage species in several xerothermic communities of the Lublin Upland were carried out in 2004 and 2005. Most plants species visited by bees are grouped in plots of the Brachypodio-Teucrietum and the Adonido-Brachypodietum pinnati communities. The nectariferous and polleniferous taxons are mostly perennials predominated by hemicryptophytes (79%), others are terophytes and geophytes (21%). Successive blooming of the nectariferous and polleniferous species in both associations ensures unbroken food flow from the early spring until the late summer and early autumn. Xerothermic swards make a valuable food potential to be important for the Apoidea before and after blooming of the main forage cultivated crops.

Pharbitidis Semen, the seed of Morning glory ( Pharbitis nil ), is a medicinal agent that has traditionally been used as a purgative in Korea. Pharbilignan C (PLC) is a dihydro[ b ]-benzofuran-type neolignan from Pharbitidis Semen, which reportedly exhibited the most potent cytotoxicity against human tumor cells. To further study the antiproliferative activity of PLC, its molecular mechanisms of action in two breast adenocarcinoma cells, MCF-7 and MDA-MB 231 cells were investigated. PLC inhibited the proliferation of MDA-MB 231 and MCF-7 cells, in order of sensitivity (IC 50 of MDA-MB 231 cells: 7.0 ± 2.0 μM). PLC induced apoptosis in MDA-MB 231 cells with down regulation of Bcl-2 and up-regulation of Bax expression. It also decreased mitochondrial membrane potential accompanied with increasing initiator caspase, caspase-9 activation and executioner caspase, caspase-3 activation. This study demonstrates that PLC inhibited proliferation of MDA-MB 231 cells by inducing apoptosis via the mitochondria-mediated intrinsic pathway. [ABSTRACT FROM AUTHOR]

CDPK kinases are a unique class of calcium sensor/responders that regulate many growth and developmental processes as well as stress responses of plants. PnCDPK1 kinase from Pharbitis nil is regulated by light and contributes to seed germination, seedling growth and flower formation. Following an earlier work in which we identified the PnCDPK1 coding sequence and a 330 bp long 3′UTR (untranslated region), we present for the first time the genomic organization of PnCDPK1 , including intron analysis and the gene copy number designation. We completed the research by identifying the 5′-flanking region of PnCDPK1 and analyzed it in silico, which led to the discovery of several cis -regulatory elements involved in light regulation, embryogenesis and seed development. The functional analysis of P. nil CDPK showed characterization of the PnCDPK1 transcript and PnCDPK protein level during seed formation and fruit maturation. The greatest amount of PnCDPK1 mRNA was present in the last stages of seed maturation. Moreover, two PnCDPK proteins of different molecular masses were discovered during fruit development, showing various protein accumulation and activity profile. The 56 kDa protein dominated in the early stages of fruit development, whereas the smaller protein (52 kDa) was prominent in the latter stages. [ABSTRACT FROM AUTHOR]

Abstract: Poor nutrition and low temperature stress treatments induced flowering in the Japanese morning glory Pharbitis nil (synonym Ipomoea nil) cv. Violet. The expression of PnFT2, one of two homologs of the floral pathway integrator gene FLOWERING LOCUS T (FT), was induced by stress, whereas the expression of both PnFT1 and PnFT2 was induced by a short-day treatment. There was no positive correlation between the flowering response and the homolog expression of another floral pathway integrator gene SUPPRESSOR OF OVEREXPRESSION OF CO1 and genes upstream of PnFT, such as CONSTANS. In another cultivar, Tendan, flowering and PnFT2 expression were not induced by poor nutrition stress. Aminooxyacetic acid (AOA), a phenylalanine ammonia-lyase inhibitor, inhibited the flowering and PnFT2 expression induced by poor nutrition stress in Violet. Salicylic acid (SA) eliminated the inhibitory effects of AOA. SA enhanced PnFT2 expression under the poor nutrition stress but not under non-stress conditions. These results suggest that SA induces PnFT2 expression, which in turn induces flowering; SA on its own, however, may not be sufficient for induction. [Copyright &y& Elsevier]

Summary: Signaling pathways, and specifically the signaling pathway of calcium, have been widely implicated in the regulation of a variety of signals in plants. Calcium-dependent protein kinases (CDPKs) are essential sensor-transducers of calcium signaling pathways, the functional characterization of which is of great interest because they play important roles during growth and in response to a wide range of environmental and developmental stimuli. Here, we report the first evidence of transient and specific elevation of PnCDPK1 transcript level and enzyme activity following conversion of a leaf bud to a flower bud, as well as participation of PnCDPK1 in evocation and flower morphogenesis in Pharbitis nil. Fluorescence microscopy immunolocalization and biochemical analysis confirmed the presence of CDPK in shoot apexes. The protein level was low in leaves, vegetative apexes and increased significantly in apexes after a flowering long-induction night. In the vegetative apex, a very weak PnCDPK1 protein signal was accumulated prominently in the zone of the ground meristem and in external layers of tissues of the cortex. After the dark treatment, the signal in cells of the ground meristem was still present, but a significantly stronger signal appeared in epidermal cells, cortex tissue, and leaf primordium. At the onset of flower meristem development, the PnCDPK1 level diverged significantly. PnCDPK1 mRNA, protein level and enzyme activity were very low at the beginning of flower bud development and gradually increased in later stages, reaching the highest level in a fully open flower. Analysis of flower organs revealed that PnCDPK1 was accumulated mainly in petals and sepals rather than in pistils and stamens. Our results clearly indicate that PnCDPK1 is developmentally regulated and may be an important component in the signal transduction pathways for flower morphogenesis. Findings from this research are important for further dissecting mechanisms of flowering and functions of CDPKs in flowering plants. [Copyright &y& Elsevier]

In this study using biochemical approaches we identified two calcium-dependent protein kinases (CDPKs) named PnCDPK52 and PnCDPK56 in soluble protein extracts from seedlings of Pharbitis nil. Both enzymes phosphorylated the specific substrate histone III-S in the presence of Ca and cross-reacted with antibodies against the CDPK. PnCDPKs exhibited quite different activity and protein levels during germination and successive stages of seedling growth. PnCDPK52 protein level was high in seeds and during germination, whereas PnCDPK56 increased in the next stages of seedling growth, being the dominant enzymes in mature seedlings, of the light- and dark-grown plant. In all cases both activity and accumulation of protein PnCDPK56 was higher in dark grown plants whereas exposure to light reduced both factors. When etiolated cotyledons were exposed to light, the activity of PnCDPK56 was reduced to the basal level within 5 h. Conversely, increasing activity of PnCDPK56 in cotyledons of green plants shifted to darkness was extremely rapid, reaching the maximum level after just 1 h of darkness and then gradually decreased. Further lengthening of the darkness to 16 h resulted in a strong increase in activity at 12 h. These data indicate that at least two isoforms of CDPK are involved in germination and seedling growth of P. nil. The differences in PnCDPKs strongly argue for the pleiotropic role of these isoforms. It seems that PnCDPK52 is associated with the germination process and PnCDPK56 with seedling growth. Moreover it suggests that activity of PnCDPK56 is controlled by light via the photoreceptor-dependent pathway. [ABSTRACT FROM AUTHOR]

PnFL-2 gene is a single copy gene that was isolated from the flower-induced cotyledons of Pharbitis nil. The deduced amino acid sequence of the PnFL-2 includes two functional domains, a single TIFY (ZIM) motif and nuclear localization signal CCT motif. The TIFY family is a novel plant specific gene involved in the regulation of diverse plant specific biological process, such as development and responses to phytochrome, implying that PnFL-2 may play a role in the mechanism of flowering induction. Subcelluar localization analysis confirmed that PnFL-2 is localized in the nucleus. Expression of PnFL-2 was induced during 16h-long inductive night, whereas accumulation of its transcripts significantly abolished by night-break, completely eliminated flowering induction of Pharbitis nil. Accumulation of its induced PnFL-2 transcripts depends on tissue, such as the cotyledons and the leaves. To study its biological functions directly, we have characterized gain of function transgenic over-expression plants for PnFL-2 in Arabidopsis. Transgenic plants that constitutively over-express PnFL-2 displayed slightly early flowering time under the long day condition as compare to those in wild type. These results indicate that PnFL-2 is potentially involved in the regulation of flowering induction in Pharbitis nil. [ABSTRACT FROM AUTHOR]

GIGANTEA (GI) is a key regulator of flowering time, which is closely related to the circadian clock function in Arabidopsis. Mutations in the GI gene cause photoperiod-insensitive flowering and altered circadian rhythms. We isolated the GI ortholog PnGI from Pharbitis (Ipomoea) nil, an absolute short-day (SD) plant. PnGI mRNA expression showed diurnal rhythms that peaked at dusk under SD and long-day (LD) conditions, and also showed robust circadian rhythms under continuous dark (DD) and continuous light (LL) conditions. Short irradiation with red light during the flower-inductive dark period did not change PnGI expression levels, suggesting that such a night break does not abolish flowering by affecting the expression of PnGI. In Pharbitis, although a single dusk signal is sufficient to induce expression of the ortholog of FLOWERING LOCUS T (PnFT1), PnGI mRNA expression was not reset by single lights-off signals. Constitutive expression of PnGI (PnGI-OX) in transgenic plants altered period length in leaf-movement rhythms under LL and affected circadian rhythms of PnFT mRNA expression under DD. PnGI-OX plants formed fewer flower buds than the wild type when one-shot darkness was given. In PnGI-OX plants, expression of PnFT1 was down-regulated, suggesting that PnGI functions as a suppressor of flowering, possibly in part through down-regulation of PnFT1. [ABSTRACT FROM AUTHOR]

Aim of the study: We performed this study to investigate the anti-cancer activity of Pharbitis nil (PN) ethanol extract which has been used for herbal medicinal treatment against diseases in East Asia. Materials and methods: We analyzed the effects of PN extract on proliferation of breast cancer cell lines, MCF-7 control vector (vec) and MCF-7 human epidermal growth factor receptor 2 (HER2) cells engineered to overexpress oncogenic HER2 via retroviral infection. We performed the proliferation assay to measure the growth rate of the cells. FACS analysis was used to analyze the cell cycle. Western blot analysis was used to investigate the effect of PN on the level and activation of intracellular molecules. Results: We found that PN extract inhibited the proliferation of both MCF-7 vec and MCF-7 HER2 cells. This growth inhibition was accompanied with the increase of sub G0/G1 apoptotic fractions. When we check the efficiency of PN on the level of intracellular signaling molecules, we found that PN extract induced the inhibition of phosphorylation of HER2 and its downstream effectors, Akt and extracellular signal-regulated kinases (ERK). Active forms of both Akt and ERK were gradually decreased in PN-treated MCF-7 vec and MCF-7 HER2 cells suggesting that the growth suppressive activity of PN is related to signaling pathway. The level of cyclin D also diminished in PN-treated both cells suggesting that PN may inhibit the growth of MCF-7 vec and MCF-7 HER2 cells by perturbing cell cycle progression. It should be noted that PN decreased the growth rate of both MCF-7 vec and MCF-7 HER2 cells without changing the level and activation of p53. Conclusion: PN extract suppressed the proliferation rate of HER-2 overexpressing MCF-7 breast cancer cells inducing apoptotic cell death in vitro. Our data demonstrates that PN extracts contain useful anti-tumor activity especially against HER2 overexpressing breast cancer. [Copyright &y& Elsevier]

The stress-sensitive short-day plant Pharbitis nil var. Kidachi flowers under a 16-h light and 8-h dark regime and non-stress conditions when grown for long periods of time. Such flowering was found to occur from the third week, and the floral buds were formed from the eighth node of the main stem. When young plants were grafted onto aged plants, the scions were induced to flower early. This flower induction by grafting was more effective when older plants were used as rootstocks. Grafting experiments using a single leaf as a donor revealed that younger leaves are more responsive to flower induction, suggesting that this age-mediated flowering response is not induced by aging or senescence of individual leaves. Rather, the plant may obtain the ability to flower as the whole plant ages. Flowering does not occur under continuous light conditions. A night break given in the 8-h dark period inhibits flowering. These results suggest that 8-h dark conditions, which are normally considered to be long-day conditions, actually correspond to short-day conditions for this plant. The 8-h dark conditions caused early flowering more efficiently in older plants. The critical dark length determined by a single treatment was 12 h in 0-week-old plants and was reduced to 6 h in 2- and 4-week-old plants. These results suggest that the critical dark length becomes shorter when plants get older. The expression of PnFT1 and PnFT2, orthologs of the flowering gene FLOWERING LOCUS T, was analyzed by reverse transcription-polymerase chain reaction revealing that the expression of PnFT at the end of dark period is correlated with flowering. [ABSTRACT FROM AUTHOR]

Calcium-dependent protein kinases (CDPKs) are central calcium signal decoders. Here we report the isolation and characterization of the Japanese Morning Glory’s ( Pharbitis nil) CDPK gene, termed PnCDPK1. The full-length cDNA of 1943 bp contains an open reading frame for PnCDPK1 consisting of 514 amino acid residues and a calculated molecular mass of 57.9 kDa. The deduced amino acid sequence suggests that this protein contains the kinase domain at the amino terminus and autoregulatory and calmodulin-like domain at the carboxy terminus. Sequence alignment indicated that PnCDPK1 shared high similarities with other CDPKs. Biochemical analyses showed that bacterially expressed recombinant protein was catalytically active and was able to phosphorylate the histone III-S in a calcium-dependent manner. Besides the identification of PnCDPK1 as a member of the CDPK family, it was shown that a transcript of PnCDPK1 was modulated during germination and seedling growth. Moreover, PnCDPK1 mRNA was present in every tested organ, including root, hypocotyl, and cotyledon of the light- and dark-grown plant; however, the higher expression level was found in dark-treated ones. The PnCDPK1 mRNA level in the cotyledons exhibited a significant increase after moving seedlings into the dark, peaking 2 and 8 h after dark exposure, and then it gradually decreased. Expression was significantly changed by exposure to red light after 8 h in the dark. These data unequivocally identify the product of the PnCDPK1 gene as a calcium-dependent protein kinase and emphasize the potential that this gene can be an element of light signal transduction involved in growth and development of P. nil. [ABSTRACT FROM AUTHOR]

Abstract: The short-day plants Pharbitis nil (synonym Ipomoea nil), var. Violet and Tendan were grown in a diluted nutrient solution or tap water for 20 days under long-day conditions. Violet plants were induced to flower and vegetative growth was inhibited, whereas Tendan plants were not induced to flower, although vegetative growth was inhibited under these conditions. The Violet plants induced to flower by poor-nutrition stress produced fertile seeds and their progeny developed normally. Defoliated Violet scions grafted onto the rootstocks of Violet or Tendan were induced to flower under poor-nutrition stress conditions, but Tendan scions grafted onto the Violet rootstocks were not induced to flower. These results indicate that a transmissible flowering stimulus is involved in the induction of flowering by poor-nutrition stress. The poor-nutrition stress-induced flowering was inhibited by aminooxyacetic acid, a phenylalanine ammonia-lyase inhibitor, and this inhibition was almost completely reversed by salicylic acid (SA). However, exogenously applied SA did not induce flowering under non-stress conditions, suggesting that SA may be necessary but not sufficient to induce flowering. PnFT2, a P. nil ortholog of the flowering gene FLOWERING LOCUS T (FT) of Arabidopsis thaliana, was expressed when the Violet plants were induced to flower by growing in tap water, but expression of PnFT1, another ortholog of FT, was not induced, suggesting the specific involvement of PnFT2 in stress-induced flowering. [Copyright &y& Elsevier]

Guanylyl cyclases (GCs) are enzymes involved in the biosynthesis of cyclic guanosine monophosphate (cGMP). Here we report the cloning and characterization of a new guanylyl cyclase, designated PnGC-1, from Pharbitis nil. This gene encodes a protein of 286 amino acids, with a calculated molecular mass of 32 kDa. The predicted amino acid sequence contains all typical features and shows high identity with known plant GCs. The GST-PnGC-1 was catalytically active in E. coli cells and the purified, recombinant PnGC-1 was able to convert GTP to cGMP in the presence of Mn2+. Moreover, the enzyme activity was strongly inhibited by a specific sGC inhibitor, NS2028, whereas in the case of nitric oxide, an animal sGC stimulator, no positive effect was observed. Besides the identification of the PnGC-1 as a guanylyl cyclase, it was shown that a transcript of PnGC-1 was present in every tested organ of the light- or dark-grown plants; however, the highest expression level was found in dark-treated plants. The PnGC-1 mRNA level in the cotyledons exhibited diurnal oscillations under short-day conditions (8/16-h photoperiod). Meanwhile, monitoring of transcript levels in cotyledons exposed to a special photoperiodic regime (24 h light of low intensity then 24 h long night with or without far-red light before the night) revealed that a stabile phytochrome is involved in this process. These data unequivocally identify the product of the PnGC-1 gene as a guanylyl cyclase and emphasize the potential that soluble GC can be an element of light signal transduction. [ABSTRACT FROM AUTHOR]

To investigate the role of distinct phytochrome pools in photoperiodic timekeeping, we characterized four phytochrome genes in the short-day plant Pharbitis nil. Each PHY gene had different photosensory properties and sensitivity to night break that inhibits flowering. During extended dark periods, PHYE, PHYB, and PHYC mRNA accumulation exhibited a circadian rhythmicity indicative of control by an endogenous clock. Phylogenetic analysis recovered four clades of angiosperm phytochrome genes, phyA, phyB, phyC, and phyE. All except the phyE dade included sequences from both monocots and eudicots. In addition, phyA is sister to phyC and phyE sister to phyB, with gymnosperm sequences sister to either the phyA-phyC dade or to the phvB-phyE dade. These results suggest that a single duplication occurred in an ancestral seed plant before the divergence of extant gymnosperms from angiosperms and that two subsequent duplications occurred in an ancestral angiosperm before the divergence of monocots from eudicots. Thus in P. nil, a multigene family with different patterns of mRNA abundance in light and darkness contributes to the total phytochrome pool: one pool is light labile (phyA), whereas the other is light stable (phyB and phyE). In addition, PHYC mRNA represents a third phytochrome pooi with intermediate photosensory properties. [ABSTRACT FROM AUTHOR]

The article presents a study which investigates the expression of PnAOC, an Allene Oxide Cyclase gene from Pharbitis nil upon theobroxide treatment to determine its role in jasmonic acid (JA) biosynthesis. It was found out that PnAOC's expression was stimulated by theobroxide. The study reveals that AOC is the most important enzyme for the biosynthesis of JA. In addition, JA's positive feedback regulation on PnAOC expression occurred through theobroxide treatment in Pharbitis nil.

Summary: Hormones are included in the essential elements that control the induction of flowering. Ethylene is thought to be a strong inhibitor of flowering in short day plants (SDPs), whereas the involvement of abscisic acid (ABA) in the regulation of flowering of plants is not well understood. The dual role of ABA in the photoperiodic flower induction of the SDP Pharbitis nil and the interaction between ABA and ethylene were examined in the present experiments. Application of ABA on the cotyledons during the inductive 16-h-long night inhibited flowering. However, ABA application on the cotyledons or the shoot apices during the subinductive 12-h-long night resulted in slight stimulation of flowering. Application of ABA also resulted in enhanced ethylene production. Whereas nordihydroguaiaretic acid (NDGA) – an ABA biosynthesis inhibitor – applied on the cotyledons of 5-d-old seedlings during the inductive night inhibited both the formation of axillary and of terminal flower buds, application of 2-aminoethoxyvinylglycine (AVG) and 2,5-norbornadiene (NBD) – inhibitors of ethylene action – reversed the inhibitory effect of ABA on flowering. ABA levels in the cotyledons of seedlings exposed to a 16-h-long inductive night markedly increased. Such an effect was not observed when the inductive night was interrupted with a 15-min-long red light pulse or when seedlings were treated at the same time with gaseous ethylene during the dark period. Lower levels of ABA were observed in seedlings treated with NDGA during the inductive night. These results may suggest that ABA plays an important role in the photoperiodic induction of flowering in P. nil seedlings, and that the inhibitory effect of ethylene on P. nil flowering inhibition may depend on its influence on the ABA level. A reversal of the inhibitory effect of ethylene on flower induction through a simultaneous treatment of induced seedlings with both ethylene and ABA strongly supports this hypothesis. [Copyright &y& Elsevier]

Abstract: Cyclic GMP acts as a chemical switch in plant cells to modulate cellular reactions. However, its metabolism has not been extensively explored and is still poorly understood. Previous experiments suggest that an endogenous cGMP system could participate in the mechanism of phytochrome controlled photoperiodic flower induction in Pharbitis nil. In order to gain further information on the role of cGMP, we have begun to study the enzyme of cGMP synthesis. In this article, the presence of the enzyme with guanylyl cyclase (GC) activity in soluble protein fractions of P. nil is reported. A large portion of the enzymatic activity is present in the cotyledons, where enzyme activity amounted to 0.45pmolcGMP/min/mg protein. The enzyme exhibited a K m 0.5mM for GTP. A plot of 1/v versus 1/[GTP] was linear and V max was 0.74pmolcGMP/min/mg protein. It was shown that the anti-sGC antibody recognise a 40kDa protein. Moreover, the NO-donor, sodium nitroprusside (SNP) and YC-1, as a NO-independent stimulator, enhanced enzyme activity. The NS 2028 (a potent GC inhibitor) treatments provoked a 3-fold reduction of the enzyme activity in comparison to the untreated fractions. Furthermore, the influence of light on GC activity was analysed. It was noted that cGMP level increased in cool white light, and darkness inhibited enzyme activity. Exposure to blue light acts to stimulate cGMP formation, whereas in red light a rapid decrease in GC activity was observed that returned to the high level when far-red light was applied after the red light treatment. The results presented in this work strongly argue that an enzyme with guanylyl cyclase activity is present in P. nil organs and its activity is controlled by light via the photoreceptors-dependent pathways. [Copyright &y& Elsevier]

Summary: Light is one of the most important environmental factors influencing the induction of flowering in plants. Light is absorbed by specific photoreceptors – the phytochromes and cryptochromes system – which fulfil a sensory and a regulatory function in the process. The absorption of light by phytochromes initiates a cascade of related biochemical events in responsive cells, and subsequently changes plant growth and development. Induction of flowering is controlled by several paths. One is triggered by the guanosine-3′:5′-cyclic monophosphate (cGMP) level. Thus, the aim of our study was to investigate the role of cGMP in phytochrome-controlled flowering. It is best to conduct such research on short-day plants because the photoperiodic reactions of only these plants are totally unequivocal. The most commonly used plant is the model short-day plant Pharbitis nil. The seedlings of P. nil were cultivated under special photoperiodic conditions: 72-h-long darkness, 24-h-long white light with low intensity and 24-h-long inductive night. Such light conditions cause a degradation of the light-labile phytochrome. Far red (FR) treatment before night causes inactivation of the remaining light-stable phytochrome. During the 24-h-long inductive darkness period, the total amount of cGMP in cotyledons underwent fluctuations, with maxima at the 4th, 8th and 14th hours. When plants were treated with FR before the long night, fluctuations were not observed. A red light pulse given after FR treatment could reverse the effect induced by FR, and the oscillation in the cGMP level was observed again. Because the intracellular level of cGMP is controlled by the opposite action of guanylyl cyclases (GCs) and phosphodiesterases (PDEs), we first tested whether accumulation of the nucleotide in P. nil tissue may be changed after treatment with a GC stimulator or PDE inhibitor. Accumulation of the nucleotide in P. nil cotyledons treated with a stimulator of cGMP synthesis (sodium nitroprusside) was markedly (approximately 80%) higher. It was highest in the presence of dipyridamole, whereas 3-isobutyl-1-methylxanthine did not significantly affect cGMP level. These results show that the analysed compounds were able to penetrate the cotyledons’ tissue, and that they influenced enzyme activity and cGMP accumulation. FR light applied at the end of the 24-h-long white light period inhibited flowering. Exogenous cGMP added on cotyledons could reverse the effect of FR, especially when the compound was applied in the first half of the long night. Flowering was also promoted by exogenous application of guanylyl cyclase activator and phosphodiesterase inhibitors, and in particular dipyridamole. The results obtained suggest that an endogenous cGMP system could participate in the mechanism of a phytochrome-controlled flowering in P. nil. [Copyright &y& Elsevier]

Abstract: α-Ketol of octadecadienoic acid (KODA, 1) has been suggested to play a role in the photoperiod-regulated flowering in Pharbitis nil. The level of 1 in cotyledons is temporarily controlled during short-day conditions. The biosynthesis of 1 is well studied in plants; however, its in vivo conversion is less understood. We have investigated this issue by studying the metabolism of exogenously-applied [U-13C]-1, [1-14C]-1, and non-labeled 1 in P. nil seedlings by the spectroscopic methods and identified six major metabolites (4–9). We have also found that the enantiomers of 1 are differentially metabolized in P. nil seedlings. [Copyright &y& Elsevier]

We present the potential involvement of Mg2+ in the flowering mechanism in the shoot apex of the short-day plant Pharbitis nil (L.). To analyze elemental distribution in shoot apical meristems, fluorescence staining methods with Mag-fluo-4 AM and Fluo-3 AM were used. The former is sensitive to both Mg2+ and Ca2+, and the latter is a specific Ca2+ indicator. When plants were grown under continuous light conditions, some cells with intensive fluorescence of Mg2+ appeared in the top layers of the shoot apical meristem. During growth in the vegetative phase, cells in the center of the top layers accumulated large amounts of Mg2+. Exposure to a single 16 h short-day treatment induced the flowering process and dramatically reduced the fluorescence associated with Mg2+ accumulation in the top layers, suggesting that Mg2+ contributes to the flower induction process. The fluorescence associated with Ca2+ did not show this distribution difference between growth phases. A night-break treatment also influenced the fluorescence pattern. It was suggested for the first time that Mg2+ plays an important role in flower induction. [ABSTRACT FROM AUTHOR]

Flower buds, cotyledons and hypocotyls of Pharbitis nil were used as plant material. Flower buds (1–2 mm long) were excised from 3-week-old plants, grown in soil. Cotyledons of 7-day-old sterile seedlings were cut into 25 mm2 squares cotyledons whereas hypocotyls were cut to 1 mm long fragments. Explants were transferred into Petri dishes containing the Murashige and Skoog medium (MS), supplemented with either BA (11 µM·L−1) alone or BA (22 µM·L−1) and NAA (0.55 µM·L−1), and different sugars: sucrose, fructose, glucose, mannose or sorbitol (autoclaved or filter-sterilized). Addition of glucose instead of sucrose to the medium stimulated the induction of callus on flower buds and cotyledonary explants, but inhibited its growth on fragments of hypocotyls. The medium supplemented with fructose (especially filter-sterilized) stimulated the development of flower elements. Organogenesis of shoots and roots on explants was also observed. Flower buds and hypocotyls were able to regenerate both organs. Addition of fructose or glucose to the medium stimulated the organogenesis of shoots, whereas root organogenesis was inhibited on all explants used. Sorbitol strongly inhibited both induction of callus and organogenesis on all explants used. [ABSTRACT FROM AUTHOR]

The purpose of the study was to determine inhibitory effect of calcium chelator; ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) on flowering of a short-day (SD) plant Pharbitis nil. It was found that 20 mM solution of EGTA applied on cotyledons of 5-d-old P. nil seedlings four hours before the start of 16-h-long induction night decreased the flowering response by 55% compared to the control plants not treated with this Ca2+ chelator. It also caused a very significant decrease of photosynthesis rate, transpiration rate and stomatal conductance both in light and darkness conditions. The results of this study confirm earlier hypothesis suggesting the effect of Ca2+ and its modulators on P. nil flowering is due to their influence on the stomata. [ABSTRACT FROM AUTHOR]

Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from NAD+ by ADP-ribosyl cyclases described for several animal cells. Pharmacological studies suggest that cADPR is an endogenous modulator of Ca2+-induced Ca2+ release channels. There is also information about the sub-micromolar concentration of cADPR in plant cells. Whether cADPR can act as a Ca2+-mobilizing intracellular messenger in plant tissue is an unresolved question. Despite the obvious importance of monitoring cADPR cellular levels under various physiological conditions in plants, its measurement has been technically difficult and requires specialized reagents. In the present study a widely applicable sensitivity assay for cADPR is described. We show that Pharbitis nil tissue from cotyledons contains a certain cADPR level. To explain the possible roles of this second messenger in photoperiodic flower induction, some physiological experiments were also performed. The exogenous applications of cADPR to Pharbitis nil plants, which were exposed to a 12-h-long subinductive night, significantly increased flowering response. Nevertheless 8-Br-cADPR inhibited flowering when these compounds were applied during a 16-h-long inductive night. The effect of ruthenium red, a calcium channel blocker and ryanodine, a calcium channel stimulator, on the photoperiodic induction of flowering was also studied. Ruthenium red, when applied before and during an inductive 16-h dark period, slightly inhibited flowering, whereas ryanodine, when applied before and during a 12-h long subinductive night, stimulated flower bud formation. We also confirmed evidence that Ca2+ ions are involved in the photoperiodic induction of flowering. Thus, the obtained results may suggest the involvement of cyclic ADPR-activated Ca2+ mobilization in the photoperiodic flower induction process in Pharbitis nil. [ABSTRACT FROM AUTHOR]

Abstract: Because tyrosinase catalyzes melanin synthesis, tyrosinase inhibitors are important in cosmetic skin-whitening. Oxidative stress contributes to skin aging and can adversely affect skin health, which means antioxidants active in skin cells may support skin health. We examined 25 traditional Chinese herbal medicines that might be useful for skin-whitening and skin health. Extracts (100μg/mL) were tested for cytotoxicity on human epidermal melanocytes (HEMn); 12 exhibited low cytotoxicity. Their effects on tyrosinase and melanin inhibitory activities and free radical scavenging activities were further assessed. Phenolic contents were evaluated using Folin–Ciocalteu reagent. Four herbs, Pharbitis nil, Sophora japonica, Spatholobus suberectus, and Morus alba, exhibited potent inhibitory effects on tyrosinase (IC50 values 24.9, 95.6, 83.9, and 78.3μg/mL, respectively). Melanin inhibition was not dose-dependent. Sophora japonica (IC50: 14.46μg/mL, 1,1-diphenyl-2-picrylhydrazyl (DPPH); 1.95μg/mL, hydroxyl radical) and Spatholobus suberectus (IC50: 10.51μg/mL, DPPH; 4.36μg/mL, hydroxyl radical) showed good antioxidative activities and high phenolic contents (255 and 189mg of gallic acid/g extract, respectively). Among active anti-tyrosinase extracts, Sophora japonica and Spatholobus suberectus were especially potent in HEMn cells in terms of free radical scavenging effects and high phenolic contents, making them the strongest candidates for cosmetic application found in the current study. [Copyright &y& Elsevier]

Summary: Theobroxide, a novel active compound isolated from a fungus, has been reported previously to induce potato tuberization and flower bud formation in Pharbitis nil under non-inductive long-day conditions. Up to date, the action mechanism of theobroxide on flower-bud induction of P. nil, however, is still unknown. In the present study, we observed a reduction of the stem length, along with the induction of flower buds, in theobroxide-treated and short-day-grown P. nil plants. Also, the results showed that flower bud formation was delayed markedly in P. nil seedlings with removal of cotyledons or exposure to night break. The suppression effect of night-break and cotyledon-removal, however, was abolished completely by spraying theobroxide. Endogenous gibberellin1/3 contents in P. nil plants treated with theobroxide or grown under short-day conditions were relatively lower, suggesting that gibberellins probably are negatively involved in theobroxide- and short-day-induced flower-bud formation of P. nil. [Copyright &y& Elsevier]