Localization and identification of phenolic compounds in Theobroma cacao L. somatic embryogenesis.

Cocoa breeders and growers continue to face the problem of high heterogeneity between individuals derived from one progeny. Vegetative propagation by somatic embryogenesis could be a way to increase genetic gains in the field. Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. This study was conducted to investigate the phenolic composition of cocoa flowers (the explants used to achieve somatic embryogenesis) and how it changes during the process, by means of histochemistry and conventional chemical techniques. In flowers, all parts contained polyphenolics but their locations were specific to the organ considered. After placing floral explants in vitro, the polyphenolic content was qualitatively modified and maintained in the calli throughout the culture process. Among the new polyphenolics, the three most abundant were isolated and characterized by 1H- and 13C-NMR. They were hydroxycinnamic acid amides: N-trans-caffeoyl-l-DOPA or clovamide, N-trans-p-coumaroyl-l-tyrosine or deoxiclovamide, and N-trans-caffeoyl-l-tyrosine. The same compounds were found also in fresh, unfermented cocoa beans. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. Given the antioxidant nature of these compounds, they could reflect the stress status of the tissues.

Plant lock and ant key: pairwise coevolution of an exclusion filter in an ant-plant mutualism.

Although observations suggest pairwise coevolution in specific ant-plant symbioses, coevolutionary processes have rarely been demonstrated. We report on, what is to the authors' knowledge, the strongest evidence yet for reciprocal adaptation of morphological characters in a species-specific ant-plant mutualism. The plant character is the prostoma, which is a small unlignified organ at the apex of the domatia in which symbiotic ants excavate an entrance hole. Each myrmecophyte in the genus Leonardoxa has evolved a prostoma with a different shape. By performing precise measurements on the prostomata of three related myrmecophytes, on their specific associated ants and on the entrance holes excavated by symbiotic ants at the prostomata, we showed that correspondence of the plant and ant traits forms a morphological and behavioural filter. We have strong evidence for coevolution between the dimensions and shape of the symbiotic ants and the prostoma in one of the three ant-Leonardoxa associations.

New applications for an old lignified element staining reagent.

The use is reported of Mirande's reagent in epifluorescence microscopy which permits a clear distinction between cellulosic and lignified tissues. Homogeneous Prespermatophytae and gymnosperm xylem appeared entirely green with Mirande's reagent under ultraviolet excitation, whereas heteroxyled angiosperm wood showed a mixed pink and blue-green colour. This coloration was due to the fluorescence of cellulose, since certain elements in dicotyledonous wood (parenchyma, fibres, xylem rays) are not entirely lignified. Monocotyledonous (Poaceae) lignin showed an intense blue fluorescence due to hydroxycinnamic acids bound to the cell wall. The method showed that lignification occurs first in the middle lamella, and later in the secondary wall of xylem cells. In addition, this staining technique proved useful in the study of lignin and suberin deposition in response to various stress factors.

Histochemical and Cytochemical Investigations of Phenols in Roots of Banana Infected by the Burrowing Nematode Radopholus similis.

ABSTRACT The burrowing nematode Radopholus similis is one of the most damaging pathogens on banana plantations. The role of phenolics in plant defense responses to the nematode was histochemically and ultrastructurally investigated in susceptible and partially resistant cultivars. Histochemical observations of healthy roots revealed that high levels of lignin, flavonoids, dopamine, cafeic esters, and ferulic acids were associated with a very low rate of nematode root penetration in the resistant cultivar. The presence of lignified and suberized layers in endodermal cells contributed to limit invasion of the vascular bundle by the pathogen. After infection, flavonoids were seen to accumulate early in walls of cells close to the nematode-migrating channel in both cultivars and in all tissues of the infected resistant roots including the vascular tissues. The labeling pattern obtained with the gold-complexed laccase and with anti-pectin monoclonal antibodies showed that phenolics were distributed in a loosened pectin-rich material surrounding the nematode. This study provides indications that constitutive phenolics in banana roots are associated with the limitation of host penetration and colonization by R. similis. Accumulation of flavonoids in response to infection was detected in the vascular tissues of susceptible plants and in all root tissues in the partially resistant plants.

Maternity and medicinal plants in Vanuatu. II. Pharmacological screening of five selected species.

An ethnobotanical survey of reproductive behaviour in Vanuatu and an extensive literature search have resulted in the selection of five plant species (Asplenium nidus, Hemigraphis reptans, Homalanthus nutans, Dysoxylum gaudichaudianum, Pemphis acidula) used for purposes relating to human reproduction in that country. Preliminary screening was carried out to identify possible oestrogenic activity in these species as well as their effects on isolated rat uteri. Dysoxylum gaudichaudianum presented the most interest due to its spasmolytic activity (musculotropic type). Its mode of action has yet to be determined.

An incremental response to high-dose therapy in multiple myeloma.

Results of conventional chemotherapy for multiple myeloma are disappointing. High-dose chemoradiotherapy with auto-transplantation is increasingly reported and some results are encouraging. We report the results of peripheral blood stem cell transplantation (PBSCT) for multiple myeloma at a single institution over a 6-year period. Forty patients, including 18 de novo patients, received debulking chemotherapy consisting of vincristine, adriamycin, and dexamethasone or methyl-prednisolone followed by stem cell mobilization with high-dose cyclophosphamide. Twenty-nine patients received PBSCT following high-dose chemoradiotherapy. Following PBSCT 92% of evaluable patients obtained at least a partial remission and 29% reached complete remission. Objective treatment responses, defined as at least a 50% reduction in serum paraprotein or marrow plasma cells, were observed following each treatment step of debulking chemotherapy, mobilization and PBSCT in 50, 42 and 71% of patients, respectively. The median overall survival from diagnosis in patients transplanted was 50 months and the median overall and progression-free survivals following transplant were 26 and 18 months, respectively. Median follow-up was 28 months. Overall treatment-related mortality was 20% but was significantly lower in de novo vs previously treated patients at 6 and 33% respectively (P = 0.027). De novo patients were more likely to obtain complete remission and had a longer overall survival following transplant but overall survival from diagnosis was similar to previously treated patients. A low serum B2M before mobilization predicted a longer progression-free survival. PBSCT needs to be considered early following diagnosis to maximise treatment response and reduce the high treatment-related mortality seen in heavily pretreated patients. In this treatment program a dose response effect in multiple myeloma was observed possibly suggesting that more intensive therapy than a single transplant may effect greater disease response.

In vitro antifungal activity of Polyporaceae against yeasts and dermatophytes.

The antifungal activity of 38 Polyporaceae sensu lato against yeasts and dermatophytes was tested in vitro by the agar dilution method. Strains were typed organisms and clinical isolates. In this first report, Pycnoporellus fulgens (Fr.) Donk was found to be the most active species against pathological fungi and showed broad-spectrum antifungal activity against yeasts (Candida albicans, Candida glabrata) and dermatophytes (Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, Microsporum gypseum, Epidermophyton floccosum). Other Polyporaceae species also showed antifungal activity, but only against dermatophytes.

A phenylethanoid glycoside from Plantago asiatica.

A new phenylethanoid glycoside, plantasioside, was isolated from the aerial parts of Plantago asiatica. The structure of plantasioside was deduced from chemical and spectral evidence to be 1',2'-[beta(3,4-dihydroxyphenyl)-alpha,beta-dioxoethanol]-6'-O- caffeoyl- O-beta-D-glucopyranoside. In addition, the structure of orobanchoside from P. depressa and P. camtschatica was revised to be 1',2'-[beta(3,4-dihydroxyphenyl)- alpha,beta-dioxoethanol]-4'-O-caffeoyl-O-alpha-L-rham- nopyranosyl-(1-->3)-O- beta-D-glucopyranoside from beta-hydroxy-[beta(3,4-dihydroxyphenyl)-ethyl]-4'-O-caffeoyl-O-alp ha-L- rhamnopyranosyl-(1-->2)-O-beta-D-glucopyranoside.

[Amanita proxima poisoning: a new cause of acute renal insufficiency]. / Intoxication par Amanita proxima: une nouvelle cause d'insuffisance rénale aiguë.

To our knowledge Amanita proxima poisoning has never been reported. Amanita proxima is a mushroom seldomy encountered, similar to a common and edible species: Amanita ovoïdae. During October 1992, we had the opportunity to care for five cases of intoxications with Amanita proxima. In all cases early digestive disorders, cytolytic hepatitis and acute renal failure were noted. Outcome was favourable for all patients within three weeks with total recovery of both renal and hepatic functions with symptomatic treatment. Temporary dialysis was required in four patients.

Caffeic glycoside esters from Jasminum nudiflorum and some related species.

Poliumoside, forsythoside B, echinacoside and arenarioside, caffeic glycoside esters, were isolated from several species of Oleaceae. The poliumoside/forsythoside ratio distinguishes Jasminum nudiflorum from J. mesnyi. Arenarioside and forsythoside B act in Forsythia species as good hybridization markers.

[Outcome of acute renal failure caused by voluntary ingestion of Cortinarius orellanus]. / Evolution d'une insuffisance rénale aiguë par ingestion volontaire de Cortinarius orellanus.

Orellanin poisoning is characterized by an acute renal failure which can be lethal if the appropriate treatment is not given. A 31-year old woman was admitted to hospital 10 days after she had deliberately ingested 2 raw carpophores of the mushroom Cortinarius orellanus. Acute renal failure (creatininaemia 1,100 mumol/l) developed, requiring 6 sessions of haemodialysis, one of plasmapheresis and the administration of diltiazem and aminoacids. Plasma and tissue assays of orellanin, the mushroom's toxin, were performed by two-dimensional thin layer chromatography. Before haemodialysis and 10 days after ingestion of the poison, the plasma contained orellanin. Eighteen months after the attempted suicide, the plasma creatinine level was 181 mumol/l.

Intoxication by Cortinarius orellanus: detection and assay of orellanine in biological fluids and renal biopsies.

A woman suffering from acute tubulo-interstitial nephritis was admitted to the hospital ten days after deliberate intoxication by ingestion of Cortinarius orellanus. Orellanine, the main toxin responsible for orellanine poisoning, was detected in biological fluids and renal biopsies. It was assayed by direct spectrofluorimetry on two-dimensional thin-layer chromatograms after specific photodecomposition into orelline. The orellanine concentration was 6.12 mg/l in the plasma (10 days after ingestion). Orellanine levels in renal biopsies were 7 micrograms per 25 mm3 of the first biopsy (13 days after ingestion) and 24 micrograms per 8 mm3 of the second biopsy (6 months later).