Hamamelis virginiana L. extract presents antimicrobial and antibiofilm effects, absence of cytotoxicity, anti-inflammatory action, and potential to fight infections through the nitric oxide production by macrophages.

The potential of H. virginiana L. was evaluated against Candida spp. (C. albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, and C. tropicalis) and bacteria (Acinetobacter baumannii, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Staphylococcus aureus, and Streptococcus mutans). Effect on murine macrophages (RAW 264.7) was also evaluated with respect to cytotoxicity and production of cytokines (IL-1ß and TNF-α) and nitric oxide (NO). The most effective concentrations of the extract were determined by microdilution broth. These concentrations were analyzed on biofilms, after 5 min or 24 h exposure. Cytotoxicity was performed by MTT assay and quantification of cytokines and NO by ELISA and Griess reagent, respectively. The extract acted against the planktonic forms and provided significant reductions of all the microbial biofilms; besides, showed no cytotoxic effect, except at 100 mg/mL, after 24 h exposure. There was cytokine production; however, a modulatory effect was observed in groups exposed to lipopolysaccharide (LPS) from E. coli. NO production was similar or higher than the control group. Thus, H. virginiana L. extract showed antimicrobial and antibiofilm effects; absence of cytotoxicity for RAW 264.7; anti-inflammatory action; and potential to fight infections through the NO production.

Elastic pinch biomechanisms can yield consistent launch speeds regardless of projectile mass.

Energetic trade-offs are particularly pertinent to bio-ballistic systems which impart energy to projectiles exclusively during launch. We investigated such trade-offs in the spring-propelled seeds of Loropetalum chinense, Hamamelis virginiana and Fortunearia sinensis. Using similar seed-shooting mechanisms, fruits of these confamilial plants (Hamamelidaceae) span an order of magnitude in spring and seed mass. We expected that as seed mass increases, launch speed decreases. Instead, launch speed was relatively constant regardless of seed mass. We tested if fruits shoot larger seeds by storing more elastic potential energy (PE). Spring mass and PE increased as seed mass increased (in order of increasing seed mass: L. chinense, H. virginiana, F. sinensis). As seed mass to spring mass ratio increased (ratios: H. virginiana = 0.50, F. sinensis = 0.65, L. chinense = 0.84), mass-specific PE storage increased. The conversion efficiency of PE to seed kinetic energy (KE) decreased with increasing fruit mass. Therefore, similar launch speeds across scales occurred because (i) larger fruits stored more PE and (ii) smaller fruits had higher mass-specific PE storage and improved PE to KE conversion. By examining integrated spring and projectile mechanics in our focal species, we revealed diverse, energetic scaling strategies relevant to spring-propelled systems navigating energetic trade-offs.

Antifungal effect of Arnica montana and Hamamelis virginiana against Candida species

Objetivo ­ Avaliar a atividade antifúngica dos extratos glicólicos de Arnica montana e Hamamelis virginiana contra cepas de Candida spp. A candidíase é uma infecção fúngica comum, portanto, a pesquisa de novos agentes antifúngicos tem sido um alvo interessante. Várias plantas apresentaram atividades biológicas e, portanto, podem ser fontes promissoras de produtos naturais com atividades an-tifúngicas. Métodos ­ As atividades antifúngicas dos extratos glicólicos foram avaliadas por meio da determinação da concentração inibitória mínima (CIM) de acordo com o protocolo M27-S3 do Clinical and Laboratory Standards Institute (2008). Resultados ­ O ex-trato glicólico de A. montana apresentou a atividade antifúngica mais forte contra C. tropicalis, com concentração inibitória mínima (CIM) de 10% v/v e concentração fungicida mínima (MFC) de 80% v/v, seguido por C. krusei e C. glabrata, com valores de MIC e MFC de 20% v/v. Além disso, avaliamos a toxicidade dos dois extratos glicólicos no modelo Galleria mellonella usando as curvas de sobre-vivência de larvas tratadas com os extratos. Nossos resultados demonstraram que os extratos glicólicos de A. montana e H. virginiana não exibiram toxicidade contra larvas de G. mellonella e demonstraram atividade antifúngica contra espécies de Candida spp. Con-clusão ­ Assim, ambos os extratos são candidatos promissores para o desenvolvimento de novos agentes antifúngicos.

Objective ­ To evaluate the antifungal activity of Arnica montana and Hamamelis virginiana glycolic extracts against Candida strains. Methods ­ The antifungal activities of glycolic extracts were investigated by determination of the minimum inhibitory concentration (MIC) according to protocol M27-S3 of Clinical and Laboratory Standards Institute (2008). Results ­ A. montana glycolic extract showed the strongest antifungal activity against C. tropicalis, with a minimum inhibitory concentration (MIC) of 10% v/v and a minimum fungicidal concentration (MFC) of 80% v/v, then C. krusei and C. glabrata, with MIC and MFC values of 20% v/v. H. virginiana glycolic extract ex-hibited stronger activity against C. albicans and C. tropicalis, with MIC and MFC values of 10% v/v, than against C. glabrata, C. krusei, and C. parapsilosis, with MIC and MFC values of 20% v/v. Moreover, we evaluated the toxicity of the two glycolic extracts in the Galleria mellonella model using the survival curves of larvae treated with the two extracts. Our results demonstrated that the glycolic extracts of A. montana and H. virginiana exhibited no toxicity against G. mellonella larvae and demonstrated antifungal activity against Candida spe-cies. Conclusion ­ Thus, both extracts are promising candidates for the development of novel antifungal agents.

The chemical constituents from twigs of Hamamelis japonica and their antiviral activities.

Three new monoterpenoid glycosides (1-3) and one new flavanol (4) along with 15 known compounds were isolated from the twig of Hamamelis japonica Sieb. et Zucc. The chemical constituent study of the twig of H. japonica has performed for the first time in the present investigation. Their structures were determined based on extensive spectroscopic methods including 1 D and 2 D NMR and CD spectra data. All isolated compounds were tested for their antiviral activities against HRV1B-, EV71-, PR8- and CVB3-infected Vero cells. Among the tested compounds, (-)-epigallocatechin 3-O-gallate exhibited the most consistent and effective antiviral activities against EV71 and PR8 infections.

Metabolomic fingerprint of Hamamelis virginiana L. gallotannins by suspect screening analysis with UHPLC-qToF and their semiquantitative evaluation.

The evolution of the regulatory framework for medical devices in the EU (Reg 2017/745) has opened the study of complex systems emerging properties. This makes necessary to identify new analytical approaches able of characterizing complex natural substrates as completely as possible. Therefore, omics approaches and advanced analytical methods for the determination of metabolite classes appear to be at the forefront to meet this need. In this perspective, a new approach based on the suspect screening was developed to detect gallotannins. Gallotannins are a class of phenols with a polymeric nature; thus, there are no pure analytical standards available for all possible structures and their quali-quantitative determination in complex natural substrates can be a challenge. A new UHPLC-qToF method was developed and used to create an "in-house tannin database" with a dual purpose: (1) as a classic list of suspects and (2) to identify core fragments common to gallotannins to have another list of putative suspects based on the common fragment. The method was validated. The application of the method to a "system of molecules" extracted from the leaves of Hamamelis virginiana L. (Witch-hazel) allowed to the characterization of a total of 29 phenols by a suspect screening approach. Therefore, 15 gallotannins were putatively annotated while another 3 were confidently identified. All the gallotannins were semiquantified according to external regression curves of gallic acid and hamamelitannin based on core fragments at m/z 125.0244 and m/z 169.0142, the building blocks of the polymers. This new method provides a practical fit-to-purpose approach for the quali-quantitative screening evaluation of gallotannins, useful for creating multivariate control charts applicable in process development of complex natural systems or in quality control. The approach is innovative, and after specific checks, it can in principle be suitable for metabolomic fingerprint analysis of gallotannins among witch-hazel extract (WHE) samples.

Unveiling the Ability of Witch Hazel (Hamamelis virginiana L.) Bark Extract to Impair Keratinocyte Inflammatory Cascade Typical of Atopic Eczema.

Hamamelis virginiana L. bark extract is a traditional remedy for skin affections, including atopic dermatitis/eczema (AD). Hamamelis preparations contain tannins, including hamamelitannin (HT), although their pharmacological role in AD is still unknown. This study aimed to study the rational for its topical use by considering the impact of crucial biomarkers on AD pathogenesis. A standardized extract (HVE) (0.5−125 µg/mL) was compared to hamamelitannin (HT), its main compound (0.5−5 µg/mL), in a model of human keratinocytes (HaCaTs), challenged with an AD-like cytokine milieu (TNF-α, IFN-γ, and IL-4). HVE inhibited the release of mediators involved in skin autoimmunity (IL-6 and IL-17C) and allergy (TSLP, IL-6, CCL26, and MMP-9) with a concentration-dependent fashion (IC50s < 25 µg/mL). The biological mechanism was ascribed, at least in part, to the impairment of the NF-κB-driven transcription. Moreover, HVE counteracted the proliferative effects of IL-4 and recovered K10, a marker of skin differentiation. Notably, HT showed activity on well-known targets of IL-4 pathway (CCL26, K10, cell proliferation). To the best of our knowledge, this work represents the first demonstration of the potential role of Hamamelis virginiana in the control of AD symptoms, such as itch and skin barrier impairment, supporting the relevance of the whole phytocomplex.

Phylogenomics AND biogeography of Castanea (chestnut) and Hamamelis (witch-hazel) - Choosing between RAD-seq and Hyb-Seq approaches.

Hyb-Seq and RAD-seq are well-established high throughput sequencing technologies that have been increasingly used for plant phylogenomic studies. Each method has its own pros and cons. The choice between them is a practical issue for plant systematists studying the evolutionary histories of biodiversity of relatively recent origins. However, few studies have compared the congruence and conflict between results from the two methods within the same group of organisms in plants. In this study, we employed RAD-seq and Hyb-Seq of Angiosperms353 genes in phylogenomic and biogeographic studies of Hamamelis (the witch-hazels) and Castanea (chestnuts), two classic examples exhibiting the well-known eastern Asian (EA) -eastern North American (ENA) disjunct distribution, and compared them side by side. Our results showed congruences in phylogenetic inference and divergence time dating between the two data sets obtained through our customized procedures of library preparation and sequence trimming, although they differed in the number of loci and informative sites, the amount of missing data, and sampling within species. We provide recommendations regarding the selection of the two methods for phylogenomic study at generic level based on fund availability and sampling scale. If funds and time are not constrained, we recommend Hyb-Seq. If funds and time are somewhat limited and sampling is large, we recommend RAD-seq. However, we found greater conflict among gene trees from the RAD-seq data due to the short sequences per locus. Therefore, species tree building and network detecting with the RAD-seq data with short RAD-seq loci (e.g., <150 bp) should avoid using analytical methods relying on gene trees of individual locus, but using site-based methods such as SVDQuartets and D-statistic method. Our phylogenetic analyses of RAD-seq and Hyb-Seq data resulted in well-resolved species relationships. Analyses of the data using the D-statistic test and PhyloNet revealed ancient introgressions in both genera. Biogeographic analyses including fossil data using total evidence-based dated tree and DEC model applying specific inter-area dispersal probabilities revealed a complicated history for each genus, indicating multiple interareal dispersals and local extinctions within and outside areas of the taxa's modern ranges in both the Paleogene and Neogene. The study demonstrates the importance of including fossil taxa for a more accurate reconstruction of biogeographic histories of taxa to understand the EA and ENA floristic disjunction. Our results support a widespread ancestral range in EA-western North America (WNA) followed by early diversification in EA and expansion to North America (NA) and Europe for Castanea and a more widespread ancestral range in EA-ENA-WNA for Hamamelis. The origins of the modern EA-ENA disjunction in both genera were suggested to be the result of vicariance from widespread ancestors in Eurasia-ENA of the mid-Miocene and in EA-NA of the late Oligocene, respectively.

A novel family of secreted insect proteins linked to plant gall development.

In an elaborate form of inter-species exploitation, many insects hijack plant development to induce novel plant organs called galls that provide the insect with a source of nutrition and a temporary home. Galls result from dramatic reprogramming of plant cell biology driven by insect molecules, but the roles of specific insect molecules in gall development have not yet been determined. Here, we study the aphid Hormaphis cornu, which makes distinctive "cone" galls on leaves of witch hazel Hamamelis virginiana. We found that derived genetic variants in the aphid gene determinant of gall color (dgc) are associated with strong downregulation of dgc transcription in aphid salivary glands, upregulation in galls of seven genes involved in anthocyanin synthesis, and deposition of two red anthocyanins in galls. We hypothesize that aphids inject DGC protein into galls and that this results in differential expression of a small number of plant genes. dgc is a member of a large, diverse family of novel predicted secreted proteins characterized by a pair of widely spaced cysteine-tyrosine-cysteine (CYC) residues, which we named BICYCLE proteins. bicycle genes are most strongly expressed in the salivary glands specifically of galling aphid generations, suggesting that they may regulate many aspects of gall development. bicycle genes have experienced unusually frequent diversifying selection, consistent with their potential role controlling gall development in a molecular arms race between aphids and their host plants.

Avaliação in vitro da atividade antimicrobiana dos extratos de Calendula officinalis, Capsicum annuum L e Hamamelis virginiana L. sobre cepas clínicas multirresistentes de Klebsiella pneumoniae e Pseudomonas aeruginosa / In vitro Evaluation of antimicrobial activity of extracts from Calendula officinalis, Capsicum annuum L and Hamamelis virginiana L. on multiresistant clinical straines of Klebsiella pneumoniae and Pseudomonas aeruginosa

O conhecimento dos efeitos dos extratos naturais de plantas vem cada vez mais se mostrando um campo promissor. Extratos de Calendula officinalis, Capsicum annuum e Hamamelis virginiana apresentam propriedades analgésicas, anti-inflamatórias, antioxidantes e potencial antimicrobiano que deve ser aprofundado. Assim, o objetivo foi avaliar ação antimicrobiana dos extratos glicólicos em culturas planctônicas e biofilmes de cepas padrão e clínicas de Klebsiella pneumoniae e Pseudomonas aeruginosa. Foi realizado o teste de microdiluição em caldo segundo Clinical and Laboratory Standards Institute, para determinação da Concentração Inibitória Mínima (CIM) e Concentração Microbicida Mínima (CMM) dos extratos. Foram realizados biofilmes monotípicos, nos tempos de contato de 5 min e 24 h, utilizando o teste de MTT. A CIM e CMM variaram de 1,56 a 50 mg/mL para todas as cepas avaliadas. C. annuum (50 mg/mL) apresentou redução de 0,25% entre 8,60% para cepa ATCC de P. aeruginosa (5 min) e em 24 h redução de 99,89% entre 99,99%, destacando-se os extratos de C. annuum (100 mg/mL) e os de C. officinalis. C. annuum (200 mg/mL) apresentou redução de 18% de K. pneumoniae (ATCC 4352). Em 24 h houve redução de 8,86%-75,74%, para o extrato de C. annuum (100 mg/mL). Os extratos apresentaram resultados mais satisfatórios quando expostos aos tratamentos de 24 horas. As cepas clínicas K1, K2 e K3 responderam ao tratamento de C. annuum (50 e 100 mg/mL) (p>0,05%). Para cepa clínica K4, os extratos de C. annuum (50 e 100 mg/mL) e C. officinalis (12,5 mg/mL) promoveram redução de viabilidade semelhante a clorexidina 2% (p>0,05%). O extrato de C. annuum (50 e 100 mg/mL), promoveu a redução de viabilidade de P1 e P2, enquanto o extrato de C. officinalis (25 e 50 mg/mL), reduziu P1 (p>0,05%). Para as cepas P3 e P4 o extrato de 50 e 100 mg/mL de C. annuum, apresentaram resultados semelhantes a clorexidina. Conclui-se que todos os extratos apresentaram ação antimicrobiana em cultura planctônica, no entanto, o extrato de C. annuum foi o que apresentou importante ação antibiofilme (24 h) sobre cepas clínicas multirresistentes de K. pneumoniae e P. aeruginosa, podendo ser considerado um potencial agente antimicrobiano.

Empirical knowledge of natural plant extracts is increasingly proving to be a promising field. Extracts of Calendula officinalis, Capsicum annuum and Hamamelis virginiana have analgesic, anti-inflammatory, antioxidant and antimicrobial potential that should be further developed. This research objective was to evaluate the antimicrobial action of glycolic extracts in planktonic cultures and biofilms of standard and clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa. The broth microdilution test was performed according to the Clinical and Laboratory Standards Institute, to determine the Minimum Inhibitory Concentration (MIC) and Minimum Microbicide Concentration (MMC) of the extracts. Tests were performed on monotypic biofilms, at contact times of 5 min and 24 h, using the MTT test. The MIC and CMM of the extracts ranged from 1.56 to 50 mg/mL for all strains evaluated. C. annuum (50 mg/mL) showed a reduction of 0.25% between 8.60% for the ATCC strain of P. aeruginosa (5 min) and in 24 h a reduction of 99.89% between 99.99%, highlighting extracts from C. annuum (100 mg/mL) and from C. officinalis. C. annuum (200 mg/mL) showed an 18% reduction in K. pneumoniae (ATCC 4352). In 24 h there was a reduction of 8.86% between 75.74% for the extract of C. annuum (100 mg/mL). The extracts showed more satisfactory results when exposed to 24-hour treatments. Clinical strains K1, K2 and K3 responded to the treatment of C. annuum (50 and 100 mg/mL) (p>0.05%). For clinical strain K4, extracts of C. annuum (50 and 100 mg/mL) and C. officinalis (12.5 mg/mL) promoted a reduction in viability similar to chlorhexidine 2% (p>0.05%). The extract of C. annuum (50 and 100 mg/mL), reduced the viability of P1 and P2, while the extract of C. officinalis (25 and 50 mg/mL), reduced P1 (p>0.05% ). For strains P3 and P4, the extract of 50 and 100 mg/mL of C. annuum showed similar results to chlorhexidine. It is concluded that all extracts showed antimicrobial action in planktonic cultures, however, the extract of C. annuum was the one that showed an important antibiofilm action (24 h) on multiresistant clinical strains of K. pneumoniae and P. aeruginosa and can be considered a potential antimicrobial agent

Extratos de romã e hamamélis: existe ação antimicrobiana sobre cepas clínicas de Acinetobacter baumannii e Klebsiella pneumoniae multirresistentes? / Pomegranate and witch hazel extracts: is there antimicrobial action on clinical strains of multiresistant Acinetobacter baumannii and Klebsiella pneumoniae?

Acinetobacter baumannii e Klebsiella pneumoniae são patógenos prioritários globais resistentes a antibióticos, portanto, é relevante encontrar métodos alternativos para controlá-los, e os fitoterápicos têm sido investigados para essa finalidade. Em vista disso, o objetivo desse estudo foi avaliar a atividade antimicrobiana e antibiofilme do extrato glicólico de Hamamelis virginiana (hamamélis) e do extrato hidroalcóolico de Punica granatum (romã) isolados e em combinação sobre cepas ATCC e clínicas de A. baumannii e K. pneumoniae. Primeiramente, foi determinada a Concentração Inibitória Mínima (CIM) e a Concentração Microbicida Mínima (CMM) pelo teste de microdiluição em caldo, segundo Clinical and Laboratory Standards Institute (CLSI), após, o potencial antimicrobiano da combinação entre esses extratos foi analisado sobre esses micro-organismos por meio da técnica "checkerboard". Posteriormente, a redução da atividade metabólica dos biofilmes foi avaliada pelo teste de MTT (contato de 5 min e 24h), para tanto a CMM e as duas concentrações acima dela foram utilizadas. A concentração correspondente a 2X a CMM dos extratos combinados foi utilizada no teste de MTT para o tratamento dos biofilmes no tempo de 5 min e 24h. Os extratos isolados e combinados apresentaram ação microbicida contra A. baumannii e K. pneumoniae na forma planctônica. O extrato de romã apresentou CMM de 1,71 a 3,43 mg/mL para A. baumannii e de 3,43 a 13,75 mg/mL para K. pneumoniae, enquanto que Hamamélis exibiu CMM de 12,5 a 25 mg/mL para A. baumannii e de 25 a 50 mg/mL para K. pneumoniae. O uso combinado dos extratos resultou em treze concentrações aditivas para A. baumannii e quinze para K. pneumoniae. A redução de viabilidade em biofilmes de A. baumannii chegou a 64,94% para o extrato de romã, e 83,45% para o extrato de Hamamélis. K. pneumoniae apresentou redução máxima de 75,66% com extrato de Romã e de 76,56% com o extrato de Hamamelis. A redução do biofilme variou conforme a concentração e cepa quando tratado com a combinação dos extratos. Com isso, pode-se concluir que os extratos apresentam importante ação antibacteriana e antibiofilme sobre os patógenos avaliados, podendo ser considerados potenciais agentes terapêuticos para o combate destes patógenos.

Pomegranate and witch hazel extracts: is there antimicrobial action on clinical strains of multiresistant Acinetobacter baumannii and Klebsiella pneumoniae? [dissertation]. São José dos Campos (SP): São Paulo State University (Unesp), Institute of Science and Technology; 2021. ABSTRACT Acinetobacter baumannii and Klebsiella pneumoniae are global priority antibiotic resistant pathogens, therefore, it is relevant to find alternative methods to control them, and herbal medicines have been investigated for this purpose. In view of this, the aim of this study was to evaluate the antimicrobial and antibiofilm activity of the glycolic extract of Hamamelis virginiana (witch hazel) and the hydroalcoholic extract of Punica granatum (pomegranate) isolated and in combination on ATCC and clinical strains of A. baumannii and K. pneumoniae. First, the Minimum Inhibitory Concentration (MIC) and the Minimum Microbicide Concentration (MMC) were determined by the broth microdilution test, according to the Clinical and Laboratory Standards Institute (CLSI), after which the antimicrobial potential of the combination of these extracts was analyzed on these microorganisms through the "checkerboard" technique. Subsequently, the reduction of the metabolic activity of biofilms was evaluated by the MTT test (5 min and 24h contact), for both the CMM and the two concentrations above it were used. The concentration corresponding to 2X the CMM of the combined extracts was used in the MTT test for the treatment of biofilms in a time of 5 min and 24h. The isolated and combined extracts showed microbicidal action against A. baumannii and K. pneumoniae in the planktonic form. The pomegranate extract showed MMC from 1.71 to 3.43 mg/mL for A. baumannii and from 3.43 to 13.75 mg/mL for K. pneumoniae, while Witch Hazel exhibited MMC from 12.5 to 25 mg µg/ml for A. baumannii and from 25 to 50 mg/ml for K. pneumoniae. The combined use of the extracts resulted in thirteen additive concentrations for A. baumannii and fifteen for K. pneumoniae. The viability reduction in A. baumannii biofilms reached 64.94% for the pomegranate extract, and 83.45% for the Witch Hazel extract. K. pneumoniae showed a maximum reduction of 75.66% with Pomegranate extract and 76.56% with Hamamelis extract. Biofilm reduction varied according to concentration and strain when treated with the combination of extracts. Thus, it can be concluded that the extracts have an important antibacterial and antibiofilm action on the evaluated pathogens, and can be considered potential therapeutic agents to combat these pathogens.

Identification of Seasonal Variations of Antibodies against PR-10-Specific Epitopes Can Be Improved Using Peptide-Phage Display.

BACKGROUND: In pollinosis patients, allergen-specific antibody titers show seasonal variations. Little is known about these variations at the epitope level. OBJECTIVES: We aimed at investigating seasonal variations on the level of allergen epitope recognition in patients with Bet v 1-related food allergy using a peptide phage display approach. METHODS: Serum samples collected over 1 year from 4 patients of the placebo arm of the birch-associated soya allergy immunotherapy trial were included. To identify epitopes from Bet v 1-related food allergens, patient sera were used in peptide phage display experiments. In silico analysis of enriched allergen-related motifs was performed. RESULTS: We identified epitope motifs related to Bet v 1 and its homologs in soya and hazelnut (Gly m 4 and Cor a 1, respectively) that were enriched in accordance with birch and hazel pollen exposure. Within several weeks after the birch pollen season peak, the pattern of identified epitope motifs differed considerably among patients. Data for amino acid preferences in homologous Bet v 1 and Cor a 1 epitope motifs identified for one of the investigated patients suggest changes in concentration or specificity of serum antibodies for the Cor a 1 epitope motif. CONCLUSIONS: Peptide phage display data suggest an impact of birch and hazel pollen exposure on the recognition pattern of Bet v 1-like allergen epitopes. Epitope-oriented analyses could provide deeper, personalized details regarding the allergen epitope recognition influenced by pollen exposure beyond the capability of current methods.

Effect of fragaria vesca, hamamelis and tormentil on the initial bacterial colonization in situ.

OBJECTIVE: The presentin situ study aims to examine the influence of the polyphenolic tea drugs fragaria vesca, hamamelis and tormentil on the initial oral bioadhesion. DESIGN: Initial biofilm formation was performed on bovine enamel slabs which were carried intraorally by 12 subjects. After 1 min of intraoral pellicle formation, the subjects rinsed with fragaria vesca, tormentil (0.8 mg/8 mL) and hamamelis (0.2 mg/8 mL) for 10 min. Tap water served as negative control, 0.2 % CHX as positive control. The investigations took place on different days (wash-out: 2 days). Afterwards, fluorescence microscopy has been performed per test solution (n = 5) and per subject (n = 12) to visualize bacterial adhesion and glucan formation (8 h oral exposition) with DAPI, ConA and BacLight. Additionally, TEM was used to visualize the pellicle ultrastructure and expectorate samples. Statistical evaluation was carried out using the Kruskal-Wallis- (p < 0.5), Mann-Whitney U test (p < 0.5) and Bonferroni-Holm-correction (p < 0.1). RESULTS: Rinsing with the polyphenolic tea extracts reduced significantly initial bacterial colonization (DAPI) compared to the negative control. There was no significant difference betweenfragaria vesca, hamamelis and tormentil. All solutions showed a reducing effect on the glucan formation. No significant difference was observed between fragaria vesca and CHX. Considerable alterations of the pellicle's ultrastructure manifested by an increase in thickness and electron density resulted from rinsing with the three polyphenolic aqueous extracts. CONCLUSIONS: Fragaria vesca, hamamelis and tormentil significantly reduce initial bioadhesion and glucan formation in situ and are therefore recommended as adjuvant antibacterial oral therapeutics.

Efficacy and tolerability of injectable collagen-containing products in comparison to trimecaine in patients with acute lumbar spine pain (Study FUTURE-MD-Back Pain).

Low back pain (LBP) represents an important subgroup of vertebrogenic pain with estimated prevalence around 80 %. Locally acting injectable collagen for topical application has recently extended the limited range of treatment options. The aim of the study was to evaluate the efficacy and safety of injectable collagen in patients with LBP. Patients suffering from LBP (< three months) were enrolled. They were administered either collagen 4 ml or trimecaine 1 % 4 ml in the form of subcutaneous paravertebral injections into eight pre-specified points (0.5 ml per each point) in the following schedule: two administrations in the first and second week, one in the third week. The pain intensity, Thomayer distance, Oswestry disability index, Lasseque test, quality of life, consumption of rescue medication and safety were evaluated. Exertional and rest pain, evaluated by a visual analogue scale, gradually decreased in both groups. Both treatments showed a statistically significant improvement in mobility and quality of life. The consumption of paracetamol as a rescue medication was significantly lower in patients treated with collagen than in the group treated with trimecaine (p=0.048). The analgesic efficacy of locally acting injectable collagen, as well as an analgesic sparing effect when compared to local anesthetics were demonstrated.

A seed flying like a bullet: ballistic seed dispersal in Chinese witch-hazel (Hamamelis mollis OLIV., Hamamelidaceae).

The fruits of Chinese witch-hazel (Hamamelis mollis, Hamamelidaceae) act as 'drying squeeze catapults', shooting their seeds several metres away. During desiccation, the exocarp shrinks and splits open, and subsequent endocarp deformation is a complex three-dimensional shape change, including formation of dehiscence lines, opening of the apical part and formation of a constriction at the middle part. Owing to the constriction forming, mechanical pressure is increasingly applied on the seed until ejection. We describe a structural latch system consisting of connective cellular structures between endocarp and seed, which break with a distinct cracking sound upon ejection. A maximum seed velocity of 12.3 m s-1, maximum launch acceleration of 19 853 m s-2 (approx. 2000g) and maximum seed rotational velocity of 25 714 min-1 were measured. We argue that miniscule morphological differences between the inner endocarp surface and seed, which features a notable ridge, are responsible for putting spin on the seed. This hypothesis is further corroborated by the observation that there is no preferential seed rotation direction among fruits. Our findings show that H. mollis has evolved similar mechanisms for stabilizing a 'shot out' seed as humans use for stabilizing rifle bullets and are discussed in an ecological (dispersal biology), biomechanical (seed ballistics) and functional-morphological (fine-tuning and morphospace of functional endocarps) contexts, and promising additional aspects for future studies are proposed.

Electrospun Membranes Based on Schizophyllan-PVOH and Hamamelis Virginiana Extract: Antimicrobial Activity Against Microorganisms of Medical Importance.

Electrospinning is a micrometric or nanometric scale fiber manufacturing technique with structural factors such as greater contact surface and pore size that allows the incorporation of biological agents in its structure, increasing their potential for medical applications. Due to the conditions required for the electrospinning process, such as high voltage, in the present work, the evaluation of the antimicrobial activity of schizophyllan-based membranes elaborated by electrospinning at 20 kV incorporated with Hamamelis virginiana was carried out against Staphylococcus aureus, Candida albicans and Pseudomonas aeruginosa. The schizophyllan production was 1.97 gL-1 from strain Schizophyllum commune ScIBL1. The conditions for the process were standardized for voltage, feed flow, and the distance from the injector to the collector. Membranes with smooth-edged fibers, diameter of 819 nm without the presence of beads were obtained. However, it was found that the membranes lost antimicrobial activity against all the microorganisms evaluated, whereas, bioassays showed that null toxicity was presented.

The potential of commercially available phytotherapeutic compounds as new photosensitizers for dental antimicrobial PDT: A photochemical and photobiological in vitro study.

The present study evaluated the effectiveness of extracts of commercially available Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum as photosensitizers in Antimicrobial Photodynamic Therapy (aPDT). Each photosensitizer (PS) was analyzed in a spectrophotometer between 350 and 750 nm to determine the ideal light source. Once the absorption bands were determined, three light sources were selected. To determine the concentration of use, the compounds were tested at different concentrations on bovine dentin samples to evaluate the risk of staining. Once the concentration was determined, the PSs were evaluated for dark toxicity and phototoxicity on fibroblast and bacteria culture. Each compound was then irradiated with each light source and evaluated for the production of reactive oxygen species (ROS). The bacterial reduction was tested on E. faecalis culture in planktonic form and on biofilm using an energy of 10 J and an Energy Density of 26 J/cm2. The tested compounds exhibited light absorption in three bands of the visible spectrum: violet (405 nm), blue (460 nm) and red (660 nm). At a 1:6 concentration, none of the compounds caused tooth staining as they did not exhibit significant toxicity in the cells or bacterial suspension. Additionally, significant ROS production was observed when the compounds were irradiated at each wavelength. When aPDT was performed on the plactonic and biofilm bacteria, significant microbial reduction was observed in both cases, reaching a reduction of up to 5Logs. In conclusion, extracts of Curcuma longa, Citrus lemon, Hamamelis virginiana and Hypericum perforatum exhibited potential for use as photosensitizing agents in aPDT.

Understanding Solar Skin Elastosis-Cause and Treatment.

Photoageing, also called actinic ageing, is the main cause of prematurely aged skin. Our expertise in elastic fibers has led us to discover a process triggered in response to ultraviolet (UV) light and which upsets the balance of elastin fibers: there is too much elastin and insufficient lysyl oxidase (LOXL1) enzyme to form functional elastic fibers. This imbalance then leads to an accumulation of nonfunctional elastin, which forms aggregates. In addition to this imbalance, UV rays also induce elafin synthesis by fibroblasts. Known to be a marker of elastotic aggregates, elafin crystallizes the elastin fibers and stimulates the formation of aggregates that cannot be naturally eliminated by the skin. We developed a Hamamelis virginiana leaf extract that was able to restore both the balance between elastin and LOXL1 and to decrease the elafin synthesis to fight and correct the damage. This specific Hamamelis virginiana extract increased LOXL1 expression by twofold and decreased elafin synthesis. As a consequence, elastic fibers became functional and aggregates of unfunctional fibers decreased. The specific Hamamelis extract activity was confirmed in vivo with decreasing wrinkles and improving skin firmness.

Botanicals With Dermatologic Properties Derived From First Nations Healing: Part 2-Plants and Algae.

INTRODUCTION: Plants and algae have played a central role in the treatment of skin conditions in both traditional First Nations healing and in modern dermatology. The objective of this study was to examine the evidence supporting the dermatological use of seaweed, witch hazel, bearberry, and mayapple. METHODS: Four plants and algae used in traditional First Nations treatments of skin disease were selected based on expert recommendations. Several databases were searched to identify relevant citations without language restrictions. RESULTS: Seaweed has potential clinical use in the treatment of acne and wrinkles and may be incorporated into biofunctional textiles. Witch hazel is an effective and well-tolerated treatment of inflammation and diaper dermatitis. Bearberry leaves contain arbutin, a skin-lightening agent that is an alternative for the treatment of hyperpigmentation. Mayapple contains podophyllotoxin, a treatment for condyloma accuminata, molluscum contagiosum, and recalcitrant palmoplantar warts. DISCUSSION: Common plants and algae are replete with bioactive agents that may have beneficial effects on the skin. Further research will open the door to new and innovative products in the future. Limitations of this study include that the scope of our study is limited to 4 plants and algae, a small sample of the breadth of plants used by First Nations for dermatological treatments.

Effectiveness of Oral Antiseptics on Tooth Biofilm: A Study in vivo.

AIM: To evaluate the effectiveness of five different mouthwashes through measurement of the plaque index. MATERIALS AND METHODS: Fifty subjects took part in this blind study, randomized into blocks of five groups according to the active ingredient of the mouthwash: CHX group (0.12% chlorhexidine gluconate), essential oils (EO) group, cetylpyridinium chloride (CPC) group, Tri group (triclosan) and Hamamelis virginiana (HV) group. All subjects were evaluated for a reduction in the bacterial plaque index at 7, 14 and 21 days. RESULTS: There was a significant reduction in the mean plaque index during the period of evaluation (p < 0.01), and the reduction during the period of evaluation was different between mouthwashes (p < 0.01). The reduction in the plaque index at the end of 21 days was, in decreasing order, CHX > EO > CPC > Tri > HV. CONCLUSION: The reduction in the plaque index during the period of evaluation was different between the types of mouth-wash. The mouthwash containing the active ingredient chlorhexidine was the most effective, followed by the essential oil, cetylpyridinium chloride, triclosan and H. virginiana.

Contact dermatitis as an adverse reaction to some topically used European herbal medicinal products - part 2: Echinacea purpurea-Lavandula angustifolia.

This review focuses on contact dermatitis as an adverse effect of a selection of topically used herbal medicinal products for which the European Medicines Agency has completed an evaluation up to the end of November 2013 and for which a Community herbal monograph has been produced. Part 2: Echinacea purpurea Moench-Lavandula angustifolia Mill.