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Bud differentiation from haploid anther callus of geranium was achieved on Murashige and Skoog medium supplemented with 0.5 mg per 1 NAA and 2.5 mg per 1 kinetin. At concentrations higher than 5 mg per 1 kinetin, malformation and tissue senescence were evident.
Unusual monounsaturated fatty acids are major constituents (greater than 80%) in seeds of Coriandrum sativum L. (coriander) and Thunbergia alata Bojer, as well as in glandular trichomes (greater than 80% derived products) of Pelargonium x hortorum (geranium). These diverged fatty acid structures are produced via distinct plastidial acyl-acyl carrier protein (ACP) desaturases. When expressed in Arabidopsis thaliana (L.) Heynh. under strong seed-specific promoters the unusual acyl-ACP desaturases resulted in accumulation of unusual monoene fatty acids at 1-15% of seed fatty acid mass. In this study, we have examined several factors that potentially limit higher production of unusual monoenes in transgenic oilseeds. (i) Immunoblots indicated that the introduced desaturases were expressed at levels equivalent to or higher than the endogenous delta9 18:0-ACP desaturase. However, the level of unusual fatty acid produced in transgenic plants was not correlated with the level of desaturase expression. (ii) The unusual desaturases were expressed in several backgrounds, including antisense 18:0-ACP desaturase plants, in fab1 mutants, and co-expressed with specialized ACP or ferredoxin isoforms. None of these experiments led to high production of expected products. (iii) No evidence was found for degradation of the unusual fatty acids during seed development. (iv) Petroselinic acid added to developing seeds was incorporated into triacylglycerol as readily as oleic acid, suggesting no major barriers to its metabolism by enzymes of glycerolipid assembly. (v) In vitro and in situ assay of acyl-ACP desaturases revealed a large discrepancy of activity when comparing unusual acyl-ACP desaturases with the endogenous delta9 18:0-ACP desaturase. The combined results, coupled with the sensitivity of acyl-ACP desaturase activity to centrifugation and low salt or detergent suggests low production of unusual monoenes in transgenic plants may be due to the lack of, or incorrect assemble of, a necessary multi-component enzyme association.
Several strains of tobacco ringspot virus (TobRV) support the replication and encapsidation of satellite tobacco ringspot virus RNA (STobRV RNA). We have compared the nucleotide sequences of four STobRV RNAs, each initially associated with a different isolate of TobRV. A STobRV RNA from a geranium isolate of TobRV and STobRV RNA from the previously analyzed budblight isolate (J.M. Buzayan, W.L. Gerlach, G. Bruening, P. Keese, and A.R. Gould, 1986, Virology 151, 186-199) differed by a single nucleotide residue substitution. STobRV RNAs from TobRV isolates 62L and NC-87 have the same 360-residue nucleotide sequence. This sequence differs from that of the 359-nucleotide residue budblight STobRV RNA principally at locations 100 through 140. The differences between the two sequences in this region are consistent with a rearrangement of blocks of nucleotide residues. The two sequences can be folded with similar patterns of base pairing. All four STobRV RNAs share a sequence of eighty 5'-terminal and of twenty 3'-terminal residues, including the 5' hydroxyl group and 2':3'-cyclic phosphodiester group.
The biotransformation of (R)-(+)- and (S)-(-)-citronellol by fungi was studied. For screening experiments, solid-phase microextraction (SPME) was used as analytical sampling technique. It was found that sporulated surface cultures of Aspergillus niger were able to convert the substrate into cis- and trans-rose oxides and nerol oxide. The relative contents in the headspace SPME extract of the three bioconversion products cis- and trans-rose oxide and nerol oxide were up to 54, 21 and 12%, respectively. Rose oxide is found in minor amounts in some essential oils, such as Bulgarian rose oil and geranium oil and contributes to its unique odor. It is one of the most important fragrance materials in perfumery in creating rosy notes. Other bioconversion products were 6-methyl-5-hepten-2-one, 6-methyl-5-hepten-2-ol, limonene, terpinolene, linalool and alpha-terpineol. These bioconversion reactions were confirmed by sporulated surface cultures on larger scale and sampling by dynamic headspace sweep and steam distillation solvent extraction. The same conversions were noticed with A. tubingensis and Penicillium roqueforti. This bioconversion was enantioselective since more of the chiral cis- than trans-rose oxide was obtained (cisitrans ratio up to 95/5). Submerged liquid cultures of P. roqueforti yielded two unidentified metabolites after conversion of citronellol (yield up to 5%). The stability and acid-catalyzed conversion of citronellol was also investigated. No chemical oxidation or auto-oxidation products were detected in acidified liquid control broths up to pH 3.5. However, when control tests were run with solid media, acid-catalyzed conversion of the substrate to small amounts of cis- and trans-rose oxides, nerol oxide, linalool and alpha-terpineol was observed at pH 3.5 and when heat treatment (steam distillation solvent extraction) was applied.
Citotoxicity (inhibition of cell division in fertilized eggs of Loxechinus albus) and general toxicity (using embryos of Artemia salina) of plants belonging to the genera Senecio, Deschampsia, Alstroemeria, Anarthrophyllum, Chloraea and Geranium were investigated.
The antimicrobial and antiplasmid activities of essential oils (orange oil, eucalyptus oil, fennel oil, geranium oil, juniper oil, peppermint oil, rosemary oil, purified turpentine oil, thyme oil, Australian tea tree oil) and of menthol, the main component of peppermint oil, were investigated. The antimicrobial activities were determined on the Gram (+) Staphylococcus epidermidis and the Gram (-) Escherichia coli F'lac K12 LE140, and on two yeast Saccharomyces cerevisiae 0425 delta/1 and 0425 52C strains. The antiplasmid activities were investigated on E. coli F'lac bacterial strain. Each of the oils exhibited antimicrobial activity and three of them antiplasmid action. The interaction of peppermint oil and menthol with the antibiotics was studied on the same bacterial strain with the checkerboard method. Peppermint oil and menthol displayed additive synergy with oxytetracycline. A new mechanism of plasmid curing was established for one of the oil components.
Antibacterial properties of aqueous and methanolic extracts of 26 medicinal plants used in Mexico to treat gastrointestinal disorders were tested against eight different species of enteropathogens: two Escherichia coli species; two Shigella sonnei species; two Shigella flexneri species; and two Salmonella sp. species. The results showed that all crude extracts exhibited antibacterial activity, at least against one of the microorganisms tested, at concentrations of 8 mg/mL or lower. The extracts from Caesalpinia pulcherria, Chiranthodendron pentadactylon, Cocos nucifera, Geranium mexicanum (aerial parts and roots), Hippocratea excelsa, and Punica granatum possessed strong antibacterial activity against most of the pathogens tested. In general, methanolic extracts were more active than aqueous extracts. Their activity was higher than chloramphenicol but did not exceed that of trimethoprim. Shigella sonnei species showed the highest susceptibility to both extracts. This is the first evaluation of these plants against bacterial pathogen isolates, which cause diarrhea and dysentery in Mexican population.
Aqueous and methanolic extracts from 26 medicinal plants used in Mexico to treat gastrointestinal disorders were screened to evaluate their antisecretory activity on cholera toxin-induced intestinal secretion in rat jejunal loops model. Extracts were tested at a dose of 300 mg/kg. From 56 samples tested, both extracts from Chiranthodendron pentadactylon, Hippocratea excelsa and Ocimum basilicum were the most potent with inhibition values ranging from 68.0 to 87.6%. On the other hand, the methanolic extract of Geranium mexicanum (aerial parts) and the aqueous extract of Bocconia frutescens showed the highest activity with inhibition values of 93.4 and 86.0%, respectively. The results obtained in this study give some scientific support to the use of the Mexican medicinal plants employed for the treatment of gastrointestinal disorders such as diarrhea.
Mexican traditional medicine uses a great variety of plants in the treatment of gastrointestinal disorders such as diarrhea. In order to understand the properties of some of their chemical constituents, three flavonoids (kaempferol, tiliroside and (-)-epicatechin) isolated from Geranium mexicanum, Cuphea pinetorum, Helianthemum glomeratum, and Rubus coriifolius, were assayed to demonstrate their in vivo antiprotozoal activity; using an experimental infection of Giardia lamblia in suckling female CD-1 mice. Compounds tested showed antigiardial activity with values of ED(50) (micromol/kg) 0.072 for (-)-epicatechin, 2.057 for kaempferol and 1.429 for tiliroside. The most active flavonoid was the (-)-epicatechin, its activity was higher than metronidazole and emetine, drugs used as positive controls. In the case of kaempferol and tiliroside their potency was close to that of the metronidazole, but far less than emetine.
Influenza infection was induced in white ICR mice by intranasal (i.n.) inoculation of the virus A/Aichi/2/68 (H3N2). The number, migration and phagocyte indices of alveolar and peritoneal macrophages (pMØ) and of blood polymorphonuclear leukocytes (PMNs), as well as the inhibition of the PMN adherence in the presence of a specific antigen were followed for 9 days after infection. The effect of the i.n. application of a polyphenol-rich extract, designated as polyphenolic complex (PC), isolated from the medicinal plant Geranium sanguineum L., on the inspected immune parameters was studied in parallel with the virological parameters of the infection, e.g. rate of mortality, mean survival time (MST), infectious lung virus titre and consolidation of the lungs. It was found that the application of PC induced a continuous 2- to 2.5-fold rise in the number of both peritoneal and alveolar macrophages (aMØ) in the infected and healthy controls. The migration of both peritoneal and aMØ increased 1.5- to 2-fold in the group of infected PC-treated animals and four to fivefold in the control group, the maximum being on day 9. PC stimulated phagocyte activities of blood PMNs in both infected and healthy mice. The leukocyte adherence inhibition (LAI) index decreased in the infected and PC-treated animals. The restoration of the suppressed functions of phagocytes in influenza virus-infected mice (VIM) was consistent with a prolongation of MST and reduction in mortality rate, infectious virus titre and lung consolidation. The immunoenhancing properties of PC apparently contribute to the overall protective effect of the plant preparation in the lethal murine experimental influenza A/Aichi infection.
This study isolated a lignan, 7,7'-dihydroxy bursehernin, from Geranium thunbergii and investigated whether or not the lignan affects the induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW264.7 macrophages stimulated with lipopolysaccharide (LPS). The gel shift analysis and luciferase reporter gene assays using the iNOS promoter and nuclear factor-kappaB (NF-kappaB) minimal promoter showed that a treatment with 7,7'-dihydroxy bursehernin reduced the reporter activities and binding of NF-kappaB to the NF-kappaB consensus sequence, while it had no effect on the nuclear translocation of p65 and the phosphorylation/degradation of I-kappaBalpha. It was reported that a few natural compounds directly suppressed the binding activity of the NF-kappaB components to DNA. The NF-kappaB binding activity was not reversed by the in vitro exposure of the nuclear extracts to 7,7'-dihydroxy bursehernin, which suggest that a metabolite(s) of 7,7'-dihydroxy bursehernin might target the binding of the NF-kappaB complex to the DNA binding domain region in the promoter region of the iNOS gene. After incubation of RAW264.7 cells with 7,7-dihydroxy bursehernin for 18h, the levels of parent compound were negligible; while a main metabolite, 4-[4-(n-hydroxy-phenyl)-2,3-dimethyl-buta-1,3-dienyl]-benzene-1,2-diol was detected in cell lysates and culture medium.
The repellent effects on nymphal stages of Ixodes ricinus L. of some plant materials have been studied in the laboratory. The plant material consisted of an ethanolic extract from Achillea millefolium L., and volatile oils of birch and/or pine tar, citronella, cloves, eucalyptus, geranium, lavender, lily of the valley and peppermint. The most pronounced effects were observed for the oils of citronella, cloves and lily of the valley. They possessed repelling activities of the same magnitude as the reference repellent DEET (N,N-diethyl-m-toluamide). Some major constituents of these oils, e.g. citronellol and geraniol (oil of citronella and lily of the valley) and eugenol (oil of cloves) showed pronounced repelling effects. This was also the case for phenethyl alcohol, a minor component in the oil from lily of the valley.
EPs 7630 is an extract of the root of the South African geranium Pelargonium sidoides. Clinical data have shown that this herbal drug preparation can be used to treat upper respiratory tract infections (URTI). The objective of this study was to investigate the impact of EPs 7630 on group A-streptococci (GAS) adhering to and invading host epithelial cells in vitro. Adhesion was assessed by a flow cytometric adhesion assay using calcein-AM-stained S. pyogenes (DSM 2071) as test organism, and HEp-2 cells and buccal epithelial cells (BEC) as substrata. For cell invasion, HEp-2 cell monolayers were infected with S. pyogenes. Intracellular bacteria were determined using a penicillin/gentamicin-protection assay. EPs 7630 was applied in therapeutically relevant concentrations between 0 and 30 microg/ml in both test systems. Compared to controls, EPs 7630 significantly reduced GAS adhesion to HEp-2 cells in a concentration-dependent manner by up to 46% (p<0.001). This was semi-quantitatively confirmed by fluorescence microscopy. Adhesion kinetics additionally indicated a specific antagonistic effect on GAS adhesion. Pre-treatment of epithelial cells or GAS with EPs 7630 showed that it targets GAS rather than HEp-2 cells. However when using BEC as substrata, GAS adhesion increased 7-fold under the influence of EPs 7630 (p<0.001). In viability assays, HEp-2 suspension contained over 90% vital cells, whereas only 10% of the BEC were viable. EPs 7630 reduced GAS invasion of HEp-2 cells significantly when the data taken at time points 60, 120, and 180 min (p=0.026) were analyzed. In conclusion, EPs 7630 develops complementary anti-infective properties: Firstly, EPs 7630 reduces bacterial adhesion to intact epithelial cells and thus protects the organism from bacterial colonization and infection/super-infection. Secondly, EPs 7630 enhances the attachment of bacteria to decaying BEC. In this way pathogens may be trapped and rendered inactive. Thirdly, the inhibition of GAS invasion of epithelial cells protects the host from microorganisms that may have evaded host defences and antibiotic treatment, and therefore prevents recurrent infections. Thus, our investigations provide a rational basis for the treatment of URTI with EPs 7630.
In this work, partition coefficients (Pwm) and solute-micelle association constants per monomer (Km/N) were measured using micellar electrokinetic chromatography in tetraborate-sodium dodecylsulfate electrolytes for 18 important plant secondary metabolites (coumarin, verbenone, camphor, eucalyptol, carvone, alpha-terpineol, linalool, jasmone, bergapten, rose oxide, geraniol, t-anethole, citronellal, citronellol, p-cymene, limonene, caryophyllene and nerol) of wide occurrence in herbal extracts and essential oils. Caryophyllene presented a retention time longer than anthracene (micelle marker) and its set of constants could not be determined accurately. Pwm and Km/N were generated by the non-linear data fitting of both partition and solute-micelle association models for the 17 solutes under consideration (caryophyllene excluded). Pwm varied from 147 (coumarin) to 13175 (limonene) while Km/N varied from 37 (coumarin) to 3721 (limonene). Under optimal conditions, the separation of the selected compounds was attempted successfully in commercialized samples of rose, anise and geranium essential oils.
Herbal medicines are now attracting attention as potential sources of cancer preventive agents. Using inhibition of tumor necrosis factor-alpha (TNF-alpha) release assay, we studied Acer nikoense, Megusurino-ki in Japanese. Inhibitory potential was found in the leaf extract, and the main active principles were identified as geraniin and corilagin. The IC(50) values for TNF-alpha release inhibition were 43 microM for geraniin and 76 microM for corilagin, whereas that for (-)-epigallocatechin gallate (EGCG), the green tea polyphenol, as control was 26 microM. Furthermore, treatment with geraniin inhibited okadaic acid tumor promotion in a two-stage carcinogenesis experiment on mouse skin. Geraniin and corilagin are present in another well-known Japanese traditional herb, Geranium thunbergii, Genno-shoko in Japanese. Considering seasonal variations of the agents and sites of cultivation of herbs, this paper reviews the significance of geraniin as a new cancer preventive agent. In addition, based on accumulated results of green tea as a cancer preventive, we review two important results with EGCG: the synergistic effects of EGCG with sulindac or tamoxifen on cancer preventive activity in PC-9 cells, and cancer prevention of intestinal tumor development in multiple intestinal neoplasia (Min) mice by cotreatment using EGCG with sulindac. We report here new findings on additional gene expression resulting from cotreatment with EGCG and sulindac in PC-9 cells compared with gene expression by EGCG alone or sulindac alone. Overall, our results indicate that, with the continuing spread of cancer chemoprevention as a fundamental medical strategy, both clinicians and researchers should take a closer look at herbal medicine.
The painting Rubens Peale with a Geranium (1801), by Rembrandt Peale, has earned a reputation as a masterpiece of early American portraiture. In recent years the painting has also been the source of controversy, because Rubens was depicted with two pairs of eyeglasses at a time when most people would not have been portrayed with spectacles at all. Scholars of American art history and ophthalmology have studied the painting and have promoted various theories for this peculiarity. A combined study of the painting, historical documentation, and optical effects in the painting, however, sheds new light on the answer to this mystery.
Concentrations of total mercury were determined by cold-vapour atomic absorption spectroscopy (CV-AAS) in 221 caps and 221 stalks of 15 species of wild growing higher fungi/mushrooms and 221 samples of corresponding soil substrate collected in 1997-98 in Manowo County, near the city of Koszalin in North-central Poland. Mean mercury concentrations in caps and stalks of the mushroom species examined and soils varied between 30+/-31 and 920+/-280, 17+/-11 and 560+/-220, and 10+/-9 and 170+/-110 ng/g dry matter, respectively. Cap to stalk mercury concentration quotients were from 1.0+/-0.4 in poison pax (Paxillus involutus) to 2.8+/-0.7 in slippery jack (Suillus luteus). Brown cort (Cortinarius malicorius), fly agaric (Amanita muscaria), orange-brown ringless amanita (A. fulva), red-aspen bolete (Leccinum rufum) and mutagen milk cap (Lactarius necator) contained the highest concentrations of mercury both in caps and stalks, and mean concentrations varied between 600+/-750 and 920+/-280 and 370+/-470 and 560+/-220 ng/g dry matter, respectively. An estimate of daily intake of mercury from mushroom consumption indicated that the flesh of edible species of mushrooms may not pose hazards to human health even at a maximum consumption rate of 28 g/day. However, it should be noted that mercury intake from other foods will augment the daily intake rates. Species such as the sickener (Russula emetica), Geranium-scented russula (R. fellea) and poison pax (P. involutus) did not concentrate mercury as evidenced from the bioconcentration factors (BCFs: concentrations in mushroom/concentration in soil substrate), which were less than 1. Similarly, red-hot milk cap (L. rufus), rickstone funnel cap (Clitocybe geotropa) and European cow bolete (S. bovinus) were observed to be weak accumulators of mercury. Fly agaric (A. muscaria) accumulated great concentrations of mercury with BCFs reaching 73+/-42 and 38+/-22 in caps and stalks, respectively. Mercury BCFs of between 4.0+/-2.3 and 23+/-25 (caps) and 2.6+/-1.9 and 14+/-12 (stalks) were noted for the other mushroom species. Relatively great concentrations of mercury in fly agaric (A. muscaria) were due to preferential uptake of this element by this species.
In the present study we evaluated the free radical scavenging action of some medicinal herbs growing in Mongolia. The aqueous extract of nine herbs Chamenerion angustifolium (Ch.ang), Equisetum arvense (Eq.arv), Gentiana decumbens (Gn.dec), Geranium pratense (Gr.pra), Lomatogonium carinthiacum (L.car), Nonea poulla (N.pl), Phodococcum vitis-idaea (Ph.v), Sphallerocarpus gracilis (Sph.gr), Stellera chamaejasme (St.cha) were used in the present experiment. The free radical scavenging action was determined in vitro and ex vivo by using electron spin resonance (ESR) spectrometer and chemiluminescence (CL) analyzer. The results showed that extracts of Ch.ang, Gn.dec, Gr.pra, L.car, N.pl, Ph.v, Sph.gr and St. cha possess strong scavenging action of 1,1-diphenyl-2-picrylhydrazyl, superoxide and hydroxyl radicals. On the other hand, the radical scavenging action of Eq.arv was low. Extracts of N.pl and Ph.v markedly inhibited the CL generated from rat liver microsomal cytochrome P450 system whereas the CL was moderately inhibited by Eq.arv, Gn.dec, Gr.pra, L.car and St.cha. The extracts of Ch.ang and Sph.gr did not decrease the CL generation. Ch.ang, Gr.pra, L.car, N.pl, Ph.v and St.cha also depressed reactive oxygen production from polymorphonuclear leukocytes stimulated by phorbol-12-myristate acetate ex vivo. Thus it was confirmed that the medicinal herbs from Mongolia possess high antioxidant potency in vitro and ex vivo.