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The medicinal plant Helleborus niger L. (Ranunculaceae) found in the European Alps has been valued as an effective remedy for numerous diseases for centuries. Recent preclinical evaluations showed pharmacologic potential of the plant extract for cancer treatment. Furthermore, several compounds extracted from H niger exhibited potent pharmacological effects. Thus, the renaissance of this old and proven medicinal herb requires better knowledge of its bioactive ingredients. A key ingredient of H niger is protoanemonin which is responsible for its burning hot taste and vesicant effect. Protoanemonin possesses antibacterial, antifungal, cytotoxic, and antimutagenic activity, and its isolation, synthesis, and preparation as a therapeutically valuable medicinal product are well elaborated. Anemonin, the dimer of protoanemonin, has antimicrobial, anti-inflammatory, and antimalarial activity, whereas (-)-ranunculin, a protoanemonin glycoside, possesses cytotoxicity and antimutagenic activity. (+)-Ranuncoside, another protoanemonin glycoside, has a spiro acetal molecular scaffold, a feature that is responsible for the biological activity of a multitude of natural products. Furthermore, the flavonoids kaempferol glycoside, quercetin glycoside, and glucosyl-phenyllactic acid have been isolated, and other phenolics with complex structures were detected as well. The main active ingredient of H niger rhizomes is the bufadienolide hellebrin. It is a steroidal cardiac diglycoside with utility for heart failure treatment. Hellebrin and its aglycon hellebrigenin are patented as lead compounds to treat cancer. Recently, additional bufadienolides as well as ecdysteroids were isolated from H niger whole plants and showed potent cytotoxicity on human cancer cell lines. Finally, various saponins have been determined by mass spectrometry, of which 4 compounds isolated from the rhizome are awaiting pharmacological assessment. This review gives an overview of the current knowledge of the isolation, synthesis, structure elucidation, derivatization, and biological activities of known H niger constituents. Furthermore, recent advances concerning the development of pharmaceuticals from these constituents and their derivatives are described. [ABSTRACT FROM AUTHOR]

Biogenic nanoarchitectured magnetic materials have drawn serious attention throughout the last decade. We have attempted the Helleborus niger flower extract functionalized and templated biogenic synthesis of Cu nanoparticles supported Fe3O4 as a likewise novel material. The plant phytomolecules were deployed as a non-toxic sustainable reductant and an outstanding capping agent to stabilize the synthesized NPs. The synthesized Cu/H.niger@Fe3O4 nanocomposite was undergone comprehensive characterizations through Fourier transformed infrared spectroscopy (FT-IR), electron microscopy (SEM and TEM), energy dispersive X-ray spectroscopy (EDX), elemental mapping, vibrating sample magnetometer (VSM), X-ray diffraction (XRD) and inductively coupled plasma (ICP) techniques. The material was catalytically explored in the synthesis of diverse pyrano[3,2-c]chromene derivatives by coupling 4-hydroxycoumarin, malononitrile and a range of aldehydes in hot water when it afforded excellent yields. Based on its core magnetism, the catalyst was easily recovered using a magnet and reused for 8 successive times without considerable loss in catalytic activity. After the chemical application, the synthesized Cu/H.niger@Fe3O4 nanocomposite was engaged in biological assays like study of anti-oxidant properties by DPPH mediated free radical scavenging test using BHT as a reference molecule. Thereafter, on having a significant IC50 value in radical scavenging assay, we extended the bio-application of the desired nanocomposite in anticancer study of A549 and H358 human lung cell lines in-vitro through MTT assay. The cell viability of malignant lung cell line reduced dose-dependently in the presence of desired nanocomposite. So, these results suggest that synthesized Cu/H.niger@Fe3O4 as a chemotherapeutic nanomaterial have a suitable anticancer activity against lung cell lines. [ABSTRACT FROM AUTHOR]

Abstract Background The therapeutic use of Helleborus niger L. is manifold due to its specific phytochemical composition. Two compound groups, the ranunculin derivates including protoanemonin and the steroidal saponins, are also associated with toxicity (genotoxicity, disintegration of membrane structures). Therefore, in vitro investigations were performed on safety aspects of a Helleborus niger aqueous fermented extract (HNE). In addition its therapeutic potential against various cancer cell lines was assessed to gain insight into the respective mechanisms of action. Methods To evaluate the safe use of HNE, Ames and hemolytic tests were carried out. Two angiogenesis assays in 2D and 3D design were conducted to assess the anti-angiogenetic potential, for which human umbilical vein endothelial cells (HUVEC) were chosen. A panel of tumor cell lines was used in 2D and 3D proliferation assays as well in the migration- and invasion-assay. All investigations were performed with HNE compared to reference substances. The 2D proliferation assay was additionally performed with isolated compounds of HNE (characteristic steroidal saponins). Results HNE did not exhibit any genotoxic potential. Concentrations up to 10 μl/ml were classified as non-hemolytic. HNE exerted anti-angiogenetic effects in HUVEC and anti-proliferative effects in five cancer cell lines, whereas hellebosaponin A and D as well macranthosid I did not show comparable effects neither singly nor in combination. Due to the inherent instability of protoanemonin in isolated form, parallel investigations with protoanemonin could not be performed. HNE (600–1000 μg/ml) inhibited the migration of certain cancer cells by > 80% such as Caki-2, DLD-1 and SK-N-SH. Conclusion HNE exhibit neither genotoxic nor hemolytic potential. The present investigations verify the anti-angiogenetic effects on HUVEC, the anti-proliferative effects and migration-inhibiting properties on tumor cells. The lower effect of the relevant steroidal saponins compared to the whole extract underlines the fact that the latter is more effective than a blend of isolated pharmacologically active components.

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.

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. [ABSTRACT FROM AUTHOR]

A new bufadienolide (1), two new bufadienolide glycosides (2 and 3), a new ecdysteroid (4), and four known compounds (5–8), were isolated from the whole plants of Helleborus niger L. (Ranunculaceae). The structures of the new compounds (1–4) were determined by spectroscopic analysis, including 2D NMR spectral data, and hydrolytic studies. Compounds 1–6 showed cytotoxicity against HL-60 human leukemia cells, A549 human lung adenocarcinoma cells, and SBC-3 human small-cell lung cancer cells, with IC50 values ranging from 0.0055 to 1.9 µM. HL-60 cells treated with either 3 or 4 showed apoptosis characteristics, such as nuclear chromatin condensation, accumulation of sub-G1 cells, and activation of caspase-3/7. [ABSTRACT FROM AUTHOR]

Helleborus niger is an evergreen species, while H. odorus is an herbaceous understorey species. They both develop flowers before the forest canopy layer closes. Their sepals remain after flowering and have multiple biological functions. To further elucidate the functions of sepals during flower development, we examined their optical and chemical properties, and the photochemical efficiency of photosystem II in the developing, flowering, and fruiting flowers. Sepals of the two species differed significantly in the contents of photosynthetic pigments and anthocyanins, but less in the UV-absorbing substances’ contents. Significant differences in photosynthetic pigment contents were also revealed within different developmental phases. The sepal potential photochemical efficiency of photosystem II was high in all developmental phases in H. odorus, whereas in H. niger, it was initially low and later increased. In the green H. odorus sepals, we obtained typical green leaf spectra with peaks in the green and NIR regions, and a low reflectance and transmittance in the UV region. On the other hand, in the white H. niger sepals in the developing and flowering phases, the response was relatively constant along the visible and NIR regions. Pigment profiles, especially chlorophylls, were shown to be important in shaping sepal optical properties, which confirms their role in light harvesting. All significant parameters together accounted for 44% and 34% of the reflectance and transmittance spectra variability, respectively. These results may contribute to the selection of Helleborus species and to a greater understanding of the ecological diversity of understorey plants in the forests.

Background: The therapeutic use of Helleborus niger L. is manifold due to its specific phytochemical composition. Two compound groups, the ranunculin derivates including protoanemonin and the steroidal saponins, are also associated with toxicity (genotoxicity, disintegration of membrane structures). Therefore, in vitro investigations were performed on safety aspects of a Helleborus niger aqueous fermented extract (HNE). In addition its therapeutic potential against various cancer cell lines was assessed to gain insight into the respective mechanisms of action. Methods: To evaluate the safe use of HNE, Ames and hemolytic tests were carried out. Two angiogenesis assays in 2D and 3D design were conducted to assess the anti-angiogenetic potential, for which human umbilical vein endothelial cells (HUVEC) were chosen. A panel of tumor cell lines was used in 2D and 3D proliferation assays as well in the migration- and invasion-assay. All investigations were performed with HNE compared to reference substances. The 2D proliferation assay was additionally performed with isolated compounds of HNE (characteristic steroidal saponins). Results: HNE did not exhibit any genotoxic potential. Concentrations up to 10 μl/ml were classified as non-hemolytic. HNE exerted anti-angiogenetic effects in HUVEC and anti-proliferative effects in five cancer cell lines, whereas hellebosaponin A and D as well macranthosid I did not show comparable effects neither singly nor in combination. Due to the inherent instability of protoanemonin in isolated form, parallel investigations with protoanemonin could not be performed. HNE (600–1000 μg/ml) inhibited the migration of certain cancer cells by > 80% such as Caki-2, DLD-1 and SK-N-SH. Conclusion: HNE exhibit neither genotoxic nor hemolytic potential. The present investigations verify the anti-angiogenetic effects on HUVEC, the anti-proliferative effects and migration-inhibiting properties on tumor cells. The lower effect of the relevant steroidal saponins compared to the whole extract underlines the fact that the latter is more effective than a blend of isolated pharmacologically active components. [ABSTRACT FROM AUTHOR]

Background: Helleborus niger L (Ranunculaceae) is used Ayurvedic and Unani systems and other herbal medicine systems. The roots of H. niger have a good medicinal value. Aims: To conduct a pharmacognostical and phytochemical study of H. niger. Materials and Methods: The pharmacognostical studies on roots including parameters such as taxonomical, macroscopic, microscopic characters, physico-chemical, ultra-violet analysis and phytochemical studies are established. Results: Macroscopically, the roots are brownish-black in colour, cylindrical in shape, feeble odour, slightly acrid taste with irregularly branched. Microscopically the root showed the presence of epidermis, air-chambers, fissure periderm, periderm, inner cortex, pith, phloem, xylem, vessels and xylem vessels. Microscopic examination of the powder showed the presence of parenchyma cells, parenchyma mass, periderm, cell inclusion, laticifer, lateral wall pith, perforation, xylem bundle and xylem elements. Ultra-violet and ordinary light analyses with different reagents were conducted to identify the drug in powder form. Physico-chemical evaluation established, Ash values - Total, acid insoluble, water soluble and sulphated ash values were 7.3%, 4.1%, 3.7% and 5.2%, respectively. Extractive values - Alcohol soluble, water soluble and ether soluble extractive values were 22.8%, 7.4% and 5.6%, respectively. Loss on drying was 3.3%. Preliminary phytochemical screening showed the presence of carbohydrate, glycoside, saponins, flavonoid, phytosterols, tannins and phenolic compounds. Conclusions: The results of the study can serve as a valuable resource of pharmacognostic and phytochemical information. This will serve as appropriate, standards for discovery of this plant material in future investigations and applications and also contribute towards establishing pharmacopoeial standards.

Helleborus niger L. is a rhizomatous, herbaceous perennial with overwintering, divided, basal leaves. The objective of the study was to investigate the influence of different levels of sucrose (10, 20, 30, 40, 50, 60, 70, and 80 g l−1) and nitrogen salts (25%, 50%, and 100% according to MS medium) as well as temperature (15°C, 20°C) on in vitro multiplication and rooting and ex vitro acclimatization of H. niger. The growth and multiplication of axillary shoots were performed on modified MS medium supplemented with various growth regulators (2iP, BAP and kinetin – each at a concentration of 1.0 mg l−1, GA3 2.5 mg l−1). For the induction of roots, the medium was supplemented with IBA 1 mg l−1 and NAA 0.1 mg l−1. Rooted plants were transplanted in a peat–perlite substrate (4:1) in a heated greenhouse for ex vitro acclimatization. The multiplication rate of H. niger shoots, in vitro rooting, and ex vitro acclimatization were strongly dependent on the sucrose/nitrogen salt relationship in the medium. The highest multiplication rate of axillary shoots (3.7) was found at a temperature of 15°C or 20°C, on the medium with cytokinins and GA3 supplemented with sucrose 20–30 g l−1 and nitrogen salts at 50%. Sucrose at a concentration of 50 g l−1 strongly stimulated the number of roots per microplant (5.8–6.0) on the media with a reduced level of nitrogen salts (25% and 50%) when the temperatures were 20°C and 15°C, respectively. The plants rooted on the media with a high sucrose/nitrogen salt ratio showed acclimatization rates which ranged from 82% to 100%. Morphological observation of plantlets revealed obvious differences in leaf shape and size and the architecture of the root system as well as differences in the developmental stages of shoots grown on media with different sucrose and nitrogen salt concentrations.

Background: Helleborus niger L (Ranunculaceae) is used Ayurvedic and Unani systems and other herbal medicine systems. The roots of H. niger have a good medicinal value. Aims: To conduct a pharmacognostical and phytochemical study of H. niger. Materials and Methods: The pharmacognostical studies on roots including parameters such as taxonomical, macroscopic, microscopic characters, physico-chemical, ultra-violet analysis and phytochemical studies are established. Results: Macroscopically, the roots are brownish-black in colour, cylindrical in shape, feeble odour, slightly acrid taste with irregularly branched. Microscopically the root showed the presence of epidermis, air-chambers, fissure periderm, periderm, inner cortex, pith, phloem, xylem, vessels and xylem vessels. Microscopic examination of the powder showed the presence of parenchyma cells, parenchyma mass, periderm, cell inclusion, laticifer, lateral wall pith, perforation, xylem bundle and xylem elements. Ultra-violet and ordinary light analyses with different reagents were conducted to identify the drug in powder form. Physico-chemical evaluation established, Ash values - Total, acid insoluble, water soluble and sulphated ash values were 7.3%, 4.1%, 3.7% and 5.2%, respectively. Extractive values - Alcohol soluble, water soluble and ether soluble extractive values were 22.8%, 7.4% and 5.6%, respectively. Loss on drying was 3.3%. Preliminary phytochemical screening showed the presence of carbohydrate, glycoside, saponins, flavonoid, phytosterols, tannins and phenolic compounds. Conclusions: The results of the study can serve as a valuable resource of pharmacognostic and phytochemical information. This will serve as appropriate, standards for discovery of this plant material in future investigations and applications and also contribute towards establishing pharmacopoeial standards. [ABSTRACT FROM AUTHOR]

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The study is based on documentation and analysis of the major visitors and pollinators of the Christmas rose (Helleborus niger L.) flowers in Slovenian naturally occurring populations. The emphasis was put on 5 groups of insects: bees, bumblebees, large flies, small dipterous flies and pollinators of minor importance. Systematic observations took place in March 2003, at two different locations: on the valley of Bohinjska Bela and on the Peca Mountain. The first location was not far from a rural area, while the second one was in an isolated area, completely in the wild. The analysis of insect activity showed that there were obvious differences in frequency of visits among the insect groups investigated, locations and time of day. In the population of Bohinjska Bela, bees were the most important pollinators, with the highest activity occurred between 10 a.m. and 11 a.m. In the isolated population in Peca the most frequent visitors were small dipterous flies. The study suggests that the Christmas rose is probably an entomophylous (the most important pollinators are insects, such as bees and flies) and predominantly cross-pollinating species. The entomophylous nature appears to be closely associated with the specific botanic characteristics of flowers. Another pollinating agent is probably wind.

Containerized seedlings of Helleborus foetidus L. (stinking hellebore), H. niger L. (Christmas rose), and H. xhybridus L. (Lenten rose) were grown under long-day conditions in controlled-environment chambers for 95 days with 9-hr days of 14, 18, 22, 26, or 30C (57, 64, 72, 79, or 86F) in factorial combination with 15-hr nights of 10, 14, 18, 22, or 26C (50, 57, 64, 72, or 79F). Long-day conditions were provided by a 3-hr night interruption. Growth of each species responded differently to day and night temperatures. Calculated maximum root, top, and total dry weight, and leaf area of H. foetidus occurred with days/nights of 20/15, 18/13, 19/14, and 18/15C (68/59, 65/55, 66/57, and 65/59F), respectively. While night temperature (NT) had no effect on root:top ratio [RTR (root dry weight + top dry weight)], RTR was greatest (0.65) with days of 22C (72F). Helleborus niger had calculated maximum root dry weight and total dry weight with days of 14C (57F) and nights of 16 and 13C (60 and 55F), respectively. Top growth of H. niger decreased linearly as NTs increased for days of 14 or 22C (57 or 72F). Day temperatures (DTs) had no effect on RTR, whereas RTR responded quadratically as NT increased with a calculated maximum RTR at nights of 19C (66F). Leaf area was maximized at days/nights of 14/10C (57/50F). At days of 22 or 26C (72 or 79F), top growth of H. xhybridus responded quadratically as NT increased with maxima occurring at nights of 18 or 17C (64 or 63F). Root dry weight responded quadratically at days of 14, 22, or 26C (57, 72, or 79F) and calculated maxima occurred with nights of 18C (64F). At days of 22 or 26C (72 or 79F), there were quadratic responses in total dry weight with calculated maximum growth of H. xhybridus at nights of 18 or 17C (64 or 63F), respectively. For days of 14, 22, or 30C (57, 72, or 86F), there were quadratic responses in RTR with greatest RTR calculated at nights of 15, 18, or 16C (59, 64, or 60F), respectively. There were quadratic responses at days of 22 or 26C (72 or 79F) for leaf area with calculated maxima at nights of 18 or 17C (64 or 63F), respectively. As DTs increased from 14 to 30C (57 to 86F) net CO2 assimilation (PN) of H. xhybridus also increased linearly whereas increased NTs had no effficient on PN. In contrast, stomatal conductance was not impacted by DT or NT. [ABSTRACT FROM AUTHOR]

Helleborus niger is an evergreen species, while H. odorus is an herbaceous understorey species. They both develop flowers before the forest canopy layer closes. Their sepals remain after flowering and have multiple biological functions. To further elucidate the functions of sepals during flower development, we examined their optical and chemical properties, and the photochemical efficiency of photosystem II in the developing, flowering, and fruiting flowers. Sepals of the two species differed significantly in the contents of photosynthetic pigments and anthocyanins, but less in the UV-absorbing substances' contents. Significant differences in photosynthetic pigment contents were also revealed within different developmental phases. The sepal potential photochemical efficiency of photosystem II was high in all developmental phases in H. odorus, whereas in H. niger, it was initially low and later increased. In the green H. odorus sepals, we obtained typical green leaf spectra with peaks in the green and NIR regions, and a low reflectance and transmittance in the UV region. On the other hand, in the white H. niger sepals in the developing and flowering phases, the response was relatively constant along the visible and NIR regions. Pigment profiles, especially chlorophylls, were shown to be important in shaping sepal optical properties, which confirms their role in light harvesting. All significant parameters together accounted for 44% and 34% of the reflectance and transmittance spectra variability, respectively. These results may contribute to the selection of Helleborus species and to a greater understanding of the ecological diversity of understorey plants in the forests. [ABSTRACT FROM AUTHOR]

Methods currently available for evaluating pollen quality in vitro include, (a) tests of germinability; (b) tests of the stainability of the vegetative cell contents; (c) tests for enzyme activity, and (d) the fluorochromatic procedure (FCR), which tests principally the integrity of the plasmalemma of the vegetative cell. Using germinability in vitro as a standard, a comparison has been made between histochemical methods of classes (b), (c) and (d) in application to various pollens, immature, mature, and treated in ways known to affect viability and membrane state. Predictably, the lowest correlation was obtained with tests of stainability. The highest was given by the FCR, which generally provided an excellent guide to potential germinability. The FCR procedure is subject to various limitations, however, (a) A high correlation between FCR and germinability can only be expected when mature, ripe pollen is used; with immature pollen, the FCR will predict excessively high potential germinability. (b) The FCR may also predict a higher potential level of pollen function than in vitro germinability when the germination medium is sub-optimal. In this situation, however, it will generally give a better guide to fertilising capacity, (c) The FCR is not a test of pollen viability. Like germinability in vitro, it can yield a negative score with pollen which is nevertheless capable of functioning. For example, false negatives will be obtained with some species if the pollen is not properly pre-conditioned by rehydration before testing, an important point in monitoring stored pollen. The paper includes a brief discussion of the rationale of pollen testing. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract: Phytochemical investigation of Helleborus niger L. (Ranunculaceae) leaf methanol extract allowed to isolate a phenolic glucoside derivative and two flavonoid glycosides characterized as phenyllactic acid 2-O-β-d-glucopyranoside (1), quercetin 3-O-2-(E-caffeoyl)-α-l-arabinopyranosyl-(1→2)-β-d-glucopyranoside-7-O-β-d-glucopyranoside (2), and kaempferol 3-O-α-l-arabinopyranosyl-(1→2)-β-d-galactopyranoside-7-O-β-d-glucopyranoside (3), respectively. Compounds 1 and 2 were isolated for the first time and their structural characterization was obtained on the basis of extensive NMR spectral studies. [Copyright &y& Elsevier]

The aerial parts of the medicinal plant Helleborus niger L. comprise a substantial number of constituents with only few of them identified so far. To expand the knowledge of its secondary metabolite profile, extracts from H. niger leaves and stems were investigated by liquid chromatography/tandem mass spectrometry (LC/MS(n) ). Specific identification strategies using LC/MS are established and discussed in detail. The leaves turned out to contain acylated and non-acylated quercetin and kaempferol oligoglycosides, protoanemonin and its precursor ranunculin, β-ecdysone, and a variety of steroidal saponins, mainly in the furostanol form. The sapogenins were elucidated as of sarsasapogenyl, diosgenyl, and macranthogenyl structures, and confirmed by comparison with the respective reference compounds. The secondary metabolite profiles were almost identical in both plant parts except that the stems lacked kaempferol derivatives and some saponins. The ranunculin derivatives and β-ecdysone were found in both plant parts. Correlations between the location of the compound groups and the plant's defense strategies are proposed. Additionally, the role of the detected secondary metabolites as protective substances against exogenic stress and as a defense against herbivores is discussed., (Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.)