Efficient Isolation and Structure Analysis of (+)-Ranuncoside, a Unique Tricyclic Spiroacetal Glycoside, from Christmas Rose (Helleborus niger L.)

The use of medicinal herbs as remedies reaches back to the Stone Age, and their importance as a source of drugs has continuously increased since then. Herbal ingredients can serve as active pharmaceuticals themselves or as lead substances for the development of synthetic pharmaceuticals with less toxicity, higher effectiveness or with new properties. To date, only 6% of the ∼600,000 plants on earth have been tested pharmacologically. Among these, the medicinal plant Helleborus niger L. (Christmas rose) is especially promising because its leaves contain ( + )-ranuncoside 1, characterized by a spiroacetal ring system, a motif which is responsible for the biological activity of a multitude of natural products. Structure-activity relationship studies of ( + )-ranuncoside 1 are lacking and no synthesis of 1 has been described yet. Therefore, we developed a protocol for the rapid and efficient isolation of 1 from the leaves of cultivated Christmas rose. Crystals of high purity were obtained that enabled us to study the stereochemistry of 1 by NMR spectroscopy in solution for the first time. The spiro configuration, the absolute stereochemistry, and the geometry of all three rings was then confirmed by x-ray structure analysis. Our data will enable future structure-activity relationship studies to assess the potential of 1 as a lead substance for the development of novel antibiotics and anticancer agents.

Ukrasna svojstva, asortiman i uzgoj kukurijeka (Helleborus spp.) zimske cvatnje

Vrste roda Helleborus se u ukrasnoj hortikulturi koriste kao vrtne biljke, lončanice za primjenu u vanjskom prostoru, kao vrste za uređenje interijera, a sve se više traže kao cvjetna vrsta za rez tijekom zime. Cilj ovog rada je opisati osnovne morfološke značajke i uvjete uzgoja vrsta roda Helleborus te dati pregled asortimana vrsta i kultivara kasnojesenske i zimske cvatnje. Pregledom literature utvrđeno je da Helleborus niger L., crni kukurijek, na tržištu postaje sve traženija ukrasna biljka, a prate ga i križanci između vrste H. orientalis i drugih vrsta ovoga roda (H. × hybridus). Iako tržišno slabo zastupljene u ukrasnoj hortikulturi se primjenjuju i druge vrste (H. viridis, H. foetidus, H. purpurascens, H. tibethanus, H. vesicarius te H. multifidus) te sve veći broj kultivara. Za cvatnju početkom zime, posebno u vrijeme Božića, najpoznatija je vrsta Helleborus niger L., a sve se više komercijaliziraju i međuvrsni križanci. Pri tome osobit značaj imaju križanci između vrsta H. niger, H. argutifolius Viv. i H. lividus Aiton. Razmnožavanje je moguće sjemenom, dijeljenjem te kulturom tkiva. Uzgaja se u uzgojnim posudama 12- 14 cm promjera u koje se sadi od 50. do 18. tjedna. Potreban mu je supstrat pH od 5,8 do 6,0, ne podnosi visoku razinu soli u tlu, a za prodaju početkom zime uzgaja se u zaštićenim prostorima od sredine listopada. Osjetljiv je na visoku vlagu, koja uz visoke temperature potencira pojavu bolesti (Fusarium, Pythium i Phytophtora).

Helleborus Niger: Systematics, Ecology, Pollination and Production Technology

The scientific monograph covers the botanical classification of the genus Helleborus L. and the Christmas rose species (Helleborus niger L.), which has undergone numerous changes throughout its botanical history. The Christmas rose is becoming increasingly important in the ornamental plant market. Its characteristic flowering in the coldest months of the year represents a great advantage over many other species of ornamental plants. As the Christmas rose does not require high temperatures to start flowering, its cultivation can be considered affordable and environmentally friendly. The supply of attractive varieties is likely to be one of the key factors that will affect its popularity in the future. The number of genetically improved (bred) varieties is limited on the market, so selective breeding of Christmas rose will become inevitable in the future. The presented results of pollination analyses, related to insect activity, are important for breeding. Observations indicate that the Christmas rose is an entomophilous and a predominantly allogamous species. Knowledge of plant ecology and cultivation technology will also be exceptionally important. In intensive cultivation, the production technology for Christmas roses is still to a large extent incomplete. The monograph includes scientific findings on the ecology and biology of the Christmas rose, its reproductive processes, and the production technology for potted plants. The monograph also presents the results of the study on the effects of beneficial microorganisms (bacteria and endomycorrhizal fungi) on the growth and development of seedlings from in vivo and in vitro environments. The results of the study indicate that biotisation is a useful technology and that inoculation with specific microorganisms has a beneficial effect on growth.

Research protocol for homeopathic treatment of congenital zika virus infection

Background The Zika virus (ZIKV) is a flavivirus and the human disease caused by this virus has been described in the Americas in 2015. ZIKV has been identified as an etiological agent of acute exanthematous disease in Brazil. In the same year, an epidemic of microcephaly with images suggestive of congenital infection raised the suspicion of a relationship between these alterations and ZIKV infection. Epidemiological and histopathological studies point to a strong relationship between prenatal Zika virus infection and microcephaly. Newborns with microcephaly, may also present auditory and visual changes, seizures and severe neurodevelopmental impairment. In 2016, the World Health Organization (WHO) declared Zika virus (ZIKV) infection a public health emergency of international concern. Aims Clinical observation in the ambulatory school of ABRAH (Brazilian Association of Recycling and Homeopathy Assistance) of patients with encephalopathy of various origins, using the medicine Helleborus niger as equalizer of the NS (Nervous System), as described in the Complex Systems of Carillo, shows improvement in motor, cognitive and seizure disorders. Based on this observation, we propose to use this method in patients with ZIKV congenital infection. Methodology To evaluate 15 patients in follow-up at the AFR (Fluminense Rehabilitation Association), through homeopathic anamnesis, clinical, diathesic, biotypological and temperamental classification. The Gross Motor Function Classification System (GMFCS) will be used. All patients will receive Helleborus niger 6 Ch daily for 6 months. Results and discussion This study was forwarded to the research ethics committee and will begin in July 2019. Will be evaluated the use of Helleborus niger in patients with congenital infection by ZIKV, as equalizer of NS. The expected result is overall neurological improvement. Conclusion Helleborus niger, probably stimulating self-regulation through uninjured neurological pathways, will promote autopoiesis and construction of new programs of action, and will enable the improvement of patients with congenital ZIKV infection.

Evidence for Glacial Refugia of the Forest Understorey Species Helleborus niger (Ranunculaceae) in the Southern as Well as in the Northern Limestone Alps

Glacial refugia of alpine and subnival biota have been intensively studied in the European Alps but the fate of forests and their understory species in that area remains largely unclear. In order to fill this gap, we aimed at disentangling the spatiotemporal diversification of disjunctly distributed black hellebore Helleborus niger (Ranunculaceae). We applied a set of phylogeographic analyses based on restriction-site associated DNA sequencing (RADseq) data and plastid DNA sequences to a range-wide sampling of populations. These analyses were supplemented with species distribution models generated for the present and the Last Glacial Maximum (LGM). We used exploratory analyses to delimit genomically coherent groups and then employed demographic modeling to reconstruct the history of these groups. We uncovered a deep split between two major genetic groups with western and eastern distribution within the Southern Limestone Alps, likely reflecting divergent evolution since the mid-Pleistocene in two glacial refugia situated along the unglaciated southern margin of the Alps. Long-term presence in the Southern Limestone Alps is also supported by high numbers of private alleles, elevated levels of nucleotide diversity and the species’ modeled distribution at the LGM. The deep genetic divergence, however, is not reflected in leaf shape variation, suggesting that the morphological discrimination of genetically divergent entities within H. niger is questionable. At a shallower level, populations from the Northern Limestone Alps are differentiated from those in the Southern Limestone Alps in both RADseq and plastid DNA data sets, reflecting the North-South disjunction within the Eastern Alps. The underlying split was dated to ca. 0.1 mya, which is well before the LGM. In the same line, explicit tests of demographic models consistently rejected the hypothesis that the partial distribution area in the Northern Limestone Alps is the result of postglacial colonization. Taken together, our results strongly support that forest understory species such as H. niger have survived the LGM in refugia situated along the southern, but also along the northern or northeastern periphery of the Alps. Being a slow migrator, the species has likely survived repeated glacial-interglacial circles in distributional stasis while the composition of the tree canopy changed in the meanwhile.

Christrose – Helleborus niger

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