In vitro genotoxic effects of four Helichrysum species in human lymphocytes cultures

Helichrysum sanguineum, Helichrysum pamphylicum, Helichrysum orientale, Helichrysum noeanum (Asteraceae) are medicinal plants. For centuries, they have been used as tea in Turkey because of their medicinal properties. So far no scientific evidence has been found in a literature survey regarding the genotoxic effects of these plants. This work evaluated the genotoxic effects on human lymphocyte cultures induced by methanol extracts of these plants, assayed in different concentrations (0.01, 0.05, 0.1, 0.5 and 1 mg/mL). According to the results, Helichrysum noeanum, Helichrysum pamphylicum and Helichrysum sanguineum induced the formation of micronuclei and decreased the mitotic and replication indexes. Helichrysum orientale did not affect these parameters, whereas Helichrysum noeanum, Helichrysum pamphylicum and Helichrysum sanguineum were clearly genotoxic. They should therefore not be used freely in alternative medicine, although their antiproliferative activity may suggest antimitotic and anticarcinogenic properties. Helichrysum orientale could be used in alternative medicine.     

Neue phloroglucin-derivate aus Helichrysum natalitium und Helichrysum bellum

Helichrysum natalitium afforded four new phloroglucinol derivatives, while from H. bellum two further compounds of this type have been isolated; H. platypterum yielded two known derivatives. The new structures were elucidated by spectroscopic methods and some chemical transformations. The chemotaxonomical situation is briefly discussed.     

Contribution to a Taxonomic Revision of the Sicilian Helichrysum Taxa by PCA Analysis of Their Essential‐Oil Compositions

The chemical profile of the essential oils in ten populations of the genus Helichrysum Mill . (Asteraceae), collected in the loci classici of the nomenclatural types of the taxa endemic to Sicily, were analyzed. Our results confirm that the analysis of secondary metabolites can be used to fingerprint wild populations of Helichrysum, the chemical profiles being coherent with the systematic arrangement of the investigated populations in three main clusters, referring to the aggregates of H. stoechas, H. rupestre, and H. italicum, all belonging to the section Stoechadina. The correct nomenclatural designation of the investigated populations is discussed and the following two new combinations are proposed: Helichrysum preslianum subsp. compactum (Guss .) Maggio, Bruno, Guarino, Senatore & Ilardi and Helichrysum panormitanum subsp. latifolium Maggio, Bruno, Guarino, Senatore & Ilardi .     

Neue diterpene und sesquiterpene aus südafrikanischen Helichrysum-arten

The investigation of several South African Helichrysum species afforded in addition to known compounds two new kaurenic acid derivatives, three new labdane derivatives, three aromadendrene derivatives and geranylterpinene. The structures are elucidated mainly by extensive ¹H NMR-studies. The chemotaxonomic importance of these findings is discussed briefly.     

A new endemic species of Helichrysum Gaertn. (Asteraceae-Inuleae) from south Anatolia

A new species of Helichrysum Gaertn., H. orbicularifolium Sümbül, R. S. Göktürk & O. D. Düşen sp. nov. , is described and illustrated. The species is restricted to the south Anatolia Province, Antalya, Turkey, growing in humid calcareous rock crevices and on limestone cliffs. The species is compared with the closely related H. chasmolycicum . © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 141 , 251–254.     

Chemical composition and in vitro antimicrobial activity of the essential oils of two Helichrysum species from Tanzania

The chemical composition of the essential oils obtained from the aerial parts of Helichrysum cymosum and H. fulgidum, from Tanzania, were analyzed by GC and GC/MS. A total of sixty-five compounds, representing 92.4% and 88.2% of the two oils, respectively, were identified. trans-Caryophyllene, caryophyllene oxide, beta-pinene, p-cymene, spathulenol and beta-bourbonene were found to be the main components. Furthermore, the oils were tested against six gram (+/-) bacteria and three pathogenic fungi. It was found that the oil of H. fulgidum exhibited significant antimicrobial activity, while the oil of H. cymosum was not active at all.     

Evaluation of antiherpesvirus-1 and genotoxic activities of Helichrysum italicum extract

The antiherpes virus-1 and genotoxic activities of diethyl ether extract from flowering tops of Helichrysum italicum (Compositae) were investigated. The extract showed significant antiviral activity at concentrations ranging from 400 to 100 microg/ml. This activity was not due to cytotoxic effect of the extract since Vero cells exhibited altered morphology or growth characteristics indicative of cytotoxic effects at higher concentration (800 microg/ml). Moreover H. italicum extract showed no DNA-damaging activity at concentrations up to 2000 microg/disk.     

Morphological and genetic evidence of contemporary intersectional hybridisation in Mediterranean Helichrysum (Asteraceae,Gnaphalieae)

Hybridisation is considered an important evolutionary phenomenon in Gnaphalieae, but contemporary hybridisation has been little explored within the tribe. Here, hybridisation between Helichrysum orientale and Helichrysum stoechas is studied at two different localities in the islands of Crete and Rhodes (Greece). Using three different types of molecular data (AFLP, nrDNA ITS sequences and cpDNA ndhF sequences) and morphological data, the aim is to provide simultaneous and direct comparisons between molecular and morphological variation among the parental species and the studied hybrid populations. AFLP profiles, ITS sequences and morphological data support the existence of hybrids at the two localities studied, shown as morphological and genetic intermediates between the parental species. Chloroplast DNA sequences show that both parental species can act either as pollen donor or as maternal parent. Fertility of hybrids is demonstrated by the viability of seeds produced by hybrids from both localities, and the detection of a backcross specimen to H. orientale. Although there is general congruence of morphological and molecular data, the analysis of morphology and ITS sequences can fail to detect backcross hybrids.     

Helichrysum nicolai (Compositae, Gnaphalieae), Systematics of a New Dwarf Local Endemic of the Cape Verde Islands, W Africa

Helichrysum nicolai from the Cape Verdean island of São Nicolau is described as a species new to science and illustrated. The systematic position in Helichrysum of this acaulescent dwarf rosette shrub with heterogamous capitula, in which the female flowers outnumber the hermaphrodite ones, is established using an ITS phylogeny based on maximum parsimony and Bayesian inference analyses. The new species is most closely related to the Canary Island endemics H. alucense and H. monogynum. The closest relative to these three species are not the other members of the genus on the mid-Atlantic archipelagos but H. somalense from the Horn of Africa region and the southern Arabian Peninsula and the widespread tropical African-S Arabian-SE Iranian H. glumaceum, to which the SE Iranian H. makranicum is found to be conspecific. The phytogeographical significance of these relationships is discussed and at least three independent colonization events of the mid-Atlantic archipelagos by Helichrysum are concluded from the molecular data available.     

Characteristic thickened cell walls of the bracts of the 'eternal flower' Helichrysum bracteatum

Background and Aims

Helichrysum bracteatum is called an ‘eternal flower’ and has large, coloured, scarious bracts. These maintain their aesthetic value without wilting or discoloration for many years. There have been no research studies of cell death or cell morphology of the scarious bract, and hence the aim of this work was to elucidate these characteristics for the bract of H. bracteatum.

Methods

DAPI (4''6-diamidino-2-phenylindol dihydrochloride) staining and fluorescence microscopy were used for observation of cell nuclei. Light microscopy (LM), transmission electron microscopy (TEM) and polarized light microscopy were used for observation of cells, including cell wall morphology.

Key Results

Cell death occurred at the bract tip during the early stage of flower development. The cell wall was the most prominent characteristic of H. bracteatum bract cells. Characteristic thickened secondary cell walls on the inside of the primary cell walls were observed in both epidermal and inner cells. In addition, the walls of all cells exhibited birefringence. Characteristic thickened secondary cell walls have orientated cellulose microfibrils as well as general secondary cell walls of the tracheary elements. For comparison, these characters were not observed in the petal and bract tissues of Chrysanthemum morifolium.

Conclusions

Bracts at anthesis are composed of dead cells. Helichrysum bracteatum bracts have characteristic thickened secondary cell walls that have not been observed in the parenchyma of any other flowers or leaves. The cells of the H. bracteatum bract differ from other tissues with secondary cell walls, suggesting that they may be a new cell type.Key words: Helichrysum bracteatum, scarious bract, secondary cell wall, primary cell wall, cell morphology, birefringence, orientated cellulose microfibrils, cell death, DAPI, transmission electron microscopy, polarized light microscopy     

Genetic variation in Mediterranean Helichrysum italicum (Asteraceae; Gnaphalieae): do disjunct populations of subsp. microphyllum have a common origin?

The yellow‐flowered everlasting daisy Helichrysum italicum (Asteraceae, Gnaphalieae) is widely distributed in the Mediterranean basin, where it grows in continuous and widespread populations in diverse open habitats. Helichrysum italicum subsp. microphyllum has a disjunct distribution in the Balearic Islands (Majorca and Dragonera), Corsica, Sardinia, Crete and Cyprus. Numerous morphological intermediates between subsp. italicum and subsp. microphyllum are known from Corsica, where the two subspecies co‐occur. The aims of the study were to investigate if subsp. microphyllum has a common origin, constituting an independent gene pool from subsp. italicum, or if the morphological differences between subsp. microphyllum and subsp. italicum have arisen independently in different locations from a common wider gene pool. Our analyses of AFLP, cpDNA sequences and morphological characters show that there is geographic structure to the genetic variation within H. italicum, with eastern and western Mediterranean groups, which do not correspond with the division into subsp. microphyllum and subsp. italicum as currently circumscribed. Local selection on quantitative trait loci provides sufficient explanation for the morphological divergence observed and is consistent with genetic data. Within the western Mediterranean group of the species we found considerable polymorphism in chloroplast DNA sequences among and within some populations. Comparison with chloroplast DNA sequences from other Helichrysum species showed that some chloroplast haplotypes are shared across species.     

Genetic characterization of hybridization between the New Zealand everlastings Helichrysum lanceolatum and Anaphalioides bellidioides (Asteraceae: Gnaphalieae)

Intergeneric hybridization between the endemic New Zealand species Helichrysum lanceolatum and Anaphalioides bellidioides (Asteraceae: Gnaphalieae) at two Banks Peninsula populations was investigated by DNA sequencing and amplified fragment length polymorphism fingerprinting. Crosses in both directions have contributed to the generation of the F1 hybrids, but in the study season hybridization was far more common with H. lanceolatum as the maternal parent. F1 hybrids have reduced, but considerable, fertility and seed collected from them includes back-crosses. Two hybrid plants sampled from the wild appear to be back-crosses to H. lanceolatum. Despite back-crossing, introgression was not demonstrated and is at most limited in scope. These results also corroborate important roles for small population size and rarity in the formation of hybrid lineages in New Zealand's everlastings.  © 2007 Landcare Research New Zealand Ltd. Journal compilation © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 89–98.     

Flavonoids in Helichrysum pamphylicum inhibit mammalian type I DNA topoisomerase

DNA topoisomerases are important targets for cancer chemotherapy. We investigated the effects of a methanolic extract of Helichrysum pamphylicum on mammalian DNA topoisomerase I via in vitro plasmid supercoil relaxation assays. The extracts manifested a considerable inhibition of the enzyme's activity in a dose-dependent manner. We also performed a HPLC analysis to identify the flavonoid content of the H. pamphylicum extract and tested the identified flavonoids; luteolin, luteolin-4-glucoside, naringenin, helichrysinA and isoquercitrin, on DNA topoisomerase I activity. The measurement of the total antioxidant capacity of the flavonoid standards suggested that the topoisomerase inhibition might be correlated with the antioxidant capacity of the plant.     

Phylogeny of South African Gnaphalieae (Asteraceae) based on two noncoding chloroplast sequences

The Gnaphalieae are a group of sunflowers that have their greatest diversity in South America, Southern Africa, and Australia. The objective of this study was to reconstruct a phylogeny of the South African Gnaphalieae using sequence data from two noncoding chloroplast DNA sequences, the trnL intron and trnL/trnF intergenic spacer. Included in this investigation are the genera of the Gnaphalieae from the African basal groups, members of the subtribes Cassiniinae, Gnaphaliinae, and Relhaniinae, and African representatives from the large Old World genus Helichrysum. Results indicate that two Gnaphaloid genera, Printzia and Callilepis, should be excluded from the Gnaphalieae. In most trees the Relhaniinae s.s. (sensu stricto) and some of the basal taxa comprise a clade that is sister to the remainder of the tribe Gnaphalieae. The Relhaniinae, which are restricted to Africa, are not a monophyletic group as presently circumscribed, nor are the South African members of Helichrysum, the Cassiniinae and Gnaphaliinae. There is general agreement between our molecular analysis and that of morphology, particularly in the terminal branches of the trees.     

Helichrysum italicum extract interferes with the production of enterotoxins by Staphylococcus aureus

AIMS: The purpose of this work was to evaluate the effect of Helichrysum italicum extract on enterotoxin (A-D) production by Staphylococcus aureus strains. METHODS AND RESULTS: The production of enterotoxins A-D in the presence or absence of H.italicum diethyl ether extract was estimated in microtiter plates using a reversed passive latex agglutination (SET-RPLA) kit (Oxoid, Basingstoke, UK). The results indicate that, in culture medium, inhibition of staphylococcal growth and enterotoxins appeared with 250-125 microg ml(-1) of the extract. Lower concentrations of the extract (62.5-31.25 microg ml(-1)) did not affect the final viable count of Staph. aureus but reduced the production of enterotoxins B and C. CONCLUSIONS: H. italicum interferes with growth and production of enterotoxins by Staph. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: There is considerable interest in the use of natural compounds as alternative methods to control undesirable pathogenic micro-organisms.     

Interactions of antibiotics and extracts of <Emphasis Type="Italic">Helichrysum pedunculatum</Emphasis> against bacteria implicated in wound infections

The effect of combinations of the crude acetone and aqueous extracts of Helichrysum pedunculatum leaves and eight antibiotics was determined by means of checkerboard and time-kill methods. In the checkerboard method, synergy of 45.8 % was observed, being independent of Gram reaction, with combinations in the aqueous extract yielding largely (18.8 %) antagonistic interactions. The time-kill assay detected synergy (45.8 %) that was also independent of Gram reaction with a potentiation of more than 3 orders of the bactericidal activity of the test antibiotics. The crude leaf extracts of H. pedunculatum could thus be considered to be potential source of a broad-spectrum antibiotic-resistance-modifying compounds.     

Three prenylated flavanoids from Helichrysum athrixiifolium

Helichrysum athrixiifolium afforded in addition to known ones three new prenylated flavanoids, one chalcone and two flavanones.     

A diterpene related to erythroxydiol from Helichrysum refluxum

From Helichrysum refluxum, in addition to known compounds, a diterpene acid related to erythroxydiol Z was isolated. The structure was elucidated by NMR techniques.     

A comparative exploration of the phytochemical profiles and bio-pharmaceutical potential of Helichrysum stoechas subsp. barrelieri extracts obtained via five extraction techniques

We endeavoured to probe into and compare the possible effect(s) of different extraction techniques (accelerated solvent extraction (ASE), microwave-assisted extraction (MAE), ultrasonication-assisted extraction (UAE), maceration, and Soxhlet extraction (SE)) on the bioactivity (antioxidant and enzyme inhibitory activities) of the aerial parts of Helichrysum stoechas subsp. barrelieri (Ten.) Nyman. Total phenolic and flavonoid contents of the extracts obtained by different extraction methods followed the order of ASE > MAE > UAE > maceration > SE. Extract obtained by ASE was the most potent radical scavenger (219.92 and 313.12 mg Trolox equivalent [TE]/g, against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), respectively) and reducing agent (927.39 and 662.87 mg TE/g, for cupric reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP), respectively). Helichrysum stoechas extract obtained by UAE (18.67 mg ethylenediaminetetraacetic equivalent [EDTAE]/g) was the most active metal chelator and inhibitor of acetylcholinesterase (4.23 mg galantamine equivalent [GALAE]/g) and butyrylcholinesterase (6.05 mg GALAE/g) cholinesterase. Extract from maceration (183.32 mg kojic acid equivalent [KAE]/g) was most active against tyrosinase while ASE extract (1.66 mmol acarbose equivalent [ACAE]/g) effectively inhibited α-glucosidase. In conclusion, data amassed herein tend to advocate for the use of non-conventional extraction techniques, namely ASE and UAE, for the extraction of bioactive secondary metabolites from H. stoechas aerial parts.     

Modifications of hydrophobicity, in vitro adherence and cellular aggregation of Streptococcus mutans by Helichrysum italicum extract

AIMS: The purpose of the present study was to examine whether sublethal concentrations of Helichrysum italicum extract could affect some of the cariogenic properties of Streptococcus mutans. METHODS AND RESULTS: We studied the antibacterial activity of H. italicum (ethanolic extract) against oral streptococci (Strep. mutans ATCC 35668, Strep. salivarius ATCC 13419 and Strep. sanguis ATCC 10556) and its influence on cell-surface hydrophobicity, in vitro sucrose-dependent adherence to glass surface and cellular aggregation of Strep. mutans. The results indicate that all streptococci were susceptible to ethanolic extract with minimum inhibitory concentration (MIC) values of 31.25-62.50 microg x ml(-1). Sub-MIC concentrations of H. italicum (7.81-31.25 microg x ml(-1)) reduced the hydrophobicity and the adherence (almost 90%) to glass surface of Strep. mutans. The aggregation in the presence of dextran T2000 was also affected. CONCLUSION: The inhibitory activity of H. italicum extract on Strep. mutans is worthy of further study. SIGNIFICANCE AND IMPACT OF THE STUDY: There is considerable interest in the use of natural compounds as alternative methods to control undesirable micro-organisms.