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The use of the therapeutic potential of medicinal plants is currently considered a physiological method of prevention and treatment, which affects the normalization of metabolic processes and the restoration of the body's functional capabilities. Medicinal products created based on plants can be used for a longer period, including in the treatment of chronic diseases. The aim of the work is to select a technology to produce tablets containing wintergreen umbrella herb liquid extract, considering the pharmaco-technological properties of the resulting dosage form. Materials and methods. The objects of the study were pipsissewa herb liquid extract and experimental model samples of mixtures of this plant substance with excipients permitted for medical use. The research on the analysis of the liquid extract and pharmaco-technological indicators of test samples of tablet mass and tablets was carried out according to the methods of the State Pharmacopoeia of Ukraine (SPhU). Determination of the optimal technology for obtaining and quantitative composition of excipients of tablets from pipsissewa herb liquid extract was carried out using mathematical design of experiments (MDE) by the method of a four-factor experiment based on the Greco-Latin square. Results. The analysis of the liquid extract of pipsissewa showed the presence of 47.0±1.0 % of extractive (dry residue) substances. It was determined that the use of Liquid-Solid technology in the technology of obtaining tablets with plant extract based on the amorphous form of magnesium aluminometasilicate (Neusilin US2) requires the use of a combined approach that will ensure the quality of the finished product. During experimental studies, the use of three interrelated production strategies was proposed. In the process of applying the proposed strategy, it was determined that microcrystalline cellulose effectively performs the function of an adsorbent and filler; the consistency of the mixtures significantly depends on the ratio of MCC 101:extract. It was investigated that the use of double granulation will allow for the increase of the amount of active ingredient in the granulate without changing the volume of MCC 101, which will allow for the achievement of a higher extract content in the finished tablets. The use of double granulation technology can also ensure uniform distribution of the active ingredient and maintain the necessary flowability of the mixture for subsequent tabletting. The use of mathematical, experimental planning in the development of double granulation (drying) technology and the composition of excipients based on studies of the fluidity of the tabletting mass; attrition, disintegration, resistance of tablets to crushing, and organoleptic indicators became the basis for the development of the composition of the medicinal product in the form of tablets containing pipsissewa liquid extract. Conclusions. The adsorption property and its influence on the tabletting process of the synthetic amorphous form of magnesium aluminometasilicate (Neusilin US2) and microcrystalline cellulose 101 (MCC 101) were determined when using these substances as a carrier for pipsissewa grass liquid extract. The possibility of conducting a double granulation process using microcrystalline cellulose 101 (MCC 101) according to a given amount of liquid extract was proven. Using mathematical planning of the experiment, the drying conditions of the granulate and the quantitative composition of excipients were selected. [ABSTRACT FROM AUTHOR]

Diseases of the kidneys and urinary tract are a common problem in people of all ages. Kidneys filter blood, removing water-soluble waste from the body, maintain water-salt balance, stabilize blood pressure and PH level. Intoxication, hypothermia, injuries and other causes lead to problems with the kidneys - inflammatory disease, urolithiasis, etc. Therefore, the development of effective herbal remedies that affect the etiopathogenic factors of diseases is urgent. The aim. Study of the profile of phenolic compounds of umbellate wintergreen, study of antioxidant and nephroprotective properties of umbellate wintergreen extract on the model of chromate-induced renal failure (chronic kidney disease) in rats. Materials and methods. Phenolic compounds were analyzed by high-performance liquid chromatography (HPLC) using a Waters e2695 Alliance HPLC system in combination with a 2998 PDA detector (Waters, Milford, MA, USA). Study of in vitro antioxidant activity by HPLC method for 50 % extract of umbellate wintergreen herb. A Waters 2695 chromatograph (Waters, USA) equipped with a Waters 996 diode-matrix detector was used. To determine the nephroprotective effect, 60 outbred sexually mature rats (males), divided into six groups, were studied. Based on the results of biochemical studies, creatinine clearance and urea clearance were calculated in experimental animals. Results. 8 phenolic compounds - apigenin, hyperoside, quercitrin, rutin, quercetin, gallic acid, guaiaverine and isoquercetin - were identified and quantified in umbellate wintergreen by HPLC. Based on the results of a preliminary assessment of the antioxidant contribution of individual phenolic compounds to the overall effectiveness of the umbellate wintergreen grass extract, a significant effect of caffeic acid derivatives and quercetin was determined. The use of Chimaphila umbellata extract in experimental animals was marked by the normalization of diuresis, a decrease in the total protein content of urine by 1.8 times. The effect of the use of umbellate wintergreen extract was expected to be dose dependent. Conclusions. The profile of umbellate wintergreen phenolic compounds was investigated by HPLC, and their antioxidant effect was determined. In the conditions of the development of renal failure in rats, the studied extract of Chimaphila umbellata improved the physical condition of the animals, reduced their mortality, improved the excretory function of the kidneys, normalized nitrogen and protein metabolism, contributed to the protection of the structure of the kidney tissue, and therefore had a positive effect on the course of nephropathy. [ABSTRACT FROM AUTHOR]

The development of new diuretics of plant origin is an actual direction. Chimaphila umbellata (L.) is a perennial herb with diuretic, astringent, analgesic and other effects; and it can treat various conditions such as edema, dropsy, etc. Pipsissewa herb helps the removal of nitrogenous and chloride salts from the body due to the content of arbutin glycoside, tannins (up to 5 %). The aim. Evaluation of the effect of pipsissewa extract on L929 cell culture. Materials and methods. Cell line L929 (fibroblasts of mouse adipose tissue) was obtained in the low-temperature bank of the Institute of Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine. Cells were cultured in DMEM medium (Bio West, France) enriched with 10 % FBS (Lonza, Germany) with 1 % antibiotic-antimycotic (Bio West, France), in a CO2 incubator (Thermo Fisher Scientific, USA) at 37 °C in an atmosphere with 5 % CO2. Determination of the minimum toxic concentration at which the cells remained alive was evaluated by morphological features (shape, monolayer integrity, adhesion to plastic). The study of the effect of pipsissewa extract on various cell functions was determined by the following methods: the ability to preserve morphological integrity -- by the phase-contrast microscopy method, energy exchange -- by the MTT test method, pinocytotic function -- by the neutral red absorption method, migratory function -- by the scratch test method, proliferative activity -- by the doubling calculation method population Results. It is proposed to use concentrations of 0.05, 0.02, 0.01, 0.005 % of pipsissewa extract for further research. After carrying out the MTT reaction, the transition of MTT to formazan was confirmed microscopically in the negative control (native cells), at PE concentrations of 0.01 % and below, and the absence of a reaction in the positive control (cells killed by ethanol) at PE concentrations above 0.02 %. When recording the parameters of the NP absorption reaction, it was determined that PE at a concentration of 0.02 % and higher sharply suppresses pinocytotic activity, despite the partial preservation of cell adhesion, reducing the concentration by two times no longer affects mitochondria. A concentration of 0.01 % reduces proliferative activity, and at a concentration of 0.005 %, no difference with the control values was found. Conclusions. When studying the assessment of the effect of pipsisewa extract on L929 cell culture, a toxic effect on these cells was established when added to the culture medium at a concentration above 0.01 %. The toxic effect had a threshold effect. Migratory and proliferative functions were the most sensitive, energy, pinocytosis and preservation of morphological integrity of cells were less sensitive [ABSTRACT FROM AUTHOR]

Wintergreen is a wild herb that grows in the hills of Nepal that can be used for extracting oil which has several medicinal values. It is being sold into premium-value markets in the USA by doTERRA Inc. doTERRA has a co-impact sourcing partnership with the NGO Choice Humanitarian, scouts the local farmers or entrepreneurs and helps them set up the production business. The difference between the price of the final product and the price that local entrepreneurs get is very high. Despite this, the venture has created a new opportunity for local farmers to make large profit as the cost of production is low. However, the business itself has several challenges from scalability and sustainability aspects. Further, the price is set by the single buyer and the market has monopsonist characteristics; hence, the producer has less bargaining power. Despite these challenges, creating a better business environment enables the export of essential oil and that can be a potential foreign currency source for country like Nepal. [ABSTRACT FROM AUTHOR]

Premise: Difficulties in comparing extremely divergent features in fully mycoheterotrophic plants with those in closely related chlorophyllous plants have complicated attempts to reveal the evolutionary patterns and processes of fully mycoheterotrophic plants. Albino mutants of partially mycoheterotrophic plants, generally observed in Orchidaceae, have provided an ideal model for investigating the evolution of mycoheterotrophy within similar genetic backgrounds. In 2018, we found a putative albino population of Pyrola (Ericaceae). Here we aimed to reveal the identity of the albino pyroloid and confirm its fully mycoheterotrophic status. Methods: To reveal the putative albino pyroloid's identity, we examined its morphology and sequenced its chloroplast DNA. In addition, we assessed the trophic status of the putative albino pyroloid by analyzing chlorophyll fluorescence, chlorophyll concentration, and natural 13C and 15N abundances. Results: We identified albino individuals as P. japonica—otherwise a partially mycoheterotrophic species. We confirmed their albino status by their considerably lower chlorophyll fluorescence and concentrations than those of sympatrically occurring chlorophyllous plants. 13C abundance in the albino individuals was significantly higher than in the green individuals of P. japonica. Conclusions: This first report of albino mutants from partially mycoheterotrophic species in angiosperms other than orchids will play a valuable role in further studies focused on mycoheterotrophy. For instance, their δ13C and δ15N values represent a reference for fully mycoheterotrophic plants in Pyrola. Our findings also indicate the strong dependence of some leafy Pyrola species on fungal C during their entire life cycle. [ABSTRACT FROM AUTHOR]

Background and Aims Pyroloids, forest sub-shrubs of the Ericaceae family, are an important model for their mixotrophic nutrition, which mixes carbon from photosynthesis and from their mycorrhizal fungi. They have medical uses but are difficult to cultivate ex situ ; in particular, their dust seeds contain undifferentiated, few-celled embryos, whose germination is normally fully supported by fungal partners. Their germination and early ontogenesis thus remain elusive. Methods An optimized in vitro cultivation system of five representatives from the subfamily Pyroloideae was developed to study the strength of seed dormancy and the effect of different media and conditions (including light, gibberellins and soluble saccharides) on germination. The obtained plants were analysed for morphological, anatomical and histochemical development. Key Results Thanks to this novel cultivation method, which breaks dormancy and achieved up to 100 % germination, leafy shoots were obtained in vitro for representatives of all pyroloid genera (Moneses, Orthilia, Pyrola and Chimaphila). In all cases, the first post-germination stage is an undifferentiated structure, from which a root meristem later emerges, well before formation of an adventive shoot. Conclusions This cultivation method can be used for further research or for ex situ conservation of pyroloid species. After strong seed dormancy is broken, the tiny globular embryo of pyroloids germinates into an intermediary zone, which is functionally convergent with the protocorm of other plants with dust seeds such as orchids. Like the orchid protocorm, this intermediary zone produces a single meristem: however, unlike orchids, which produce a shoot meristem, pyroloids first generate a root meristem. [ABSTRACT FROM AUTHOR]

Abstract: Based on the DNA sequencing reads obtained using 454 pyrosequencing, primers amplifying 16 microsatellite loci were developed for the endangered semi‐shrub Chimaphila umbellata, which occurs sporadically in the Japanese Archipelago. These 16 loci were polymorphic in the populations sampled from the Hokkaido and Tohoku Districts; the mean number of alleles was 3.31 and 3.44, and the mean expected heterozygosity was 0.42 and 0.44, respectively. These loci were not linked to each other and contained no null alleles. Amplification using these primers was also tested in the congeneric species C. japonica, but only three of them successfully amplified DNA of the species. These markers will be used to examine genetic diversity and genetic differentiation in populations of C. umbellata. [ABSTRACT FROM AUTHOR]

Leaves under stressful conditions usually show downregulated maximum quantum efficiency of photosystem II [inferred from variable to maximum chlorophyll (Chl) a fluorescence (F/F), usually lower than 0.8], indicating photoinhibition. The usual method to evaluate the degree of photoinhibition in winter red leaves is generally by measuring the F/F on the red adaxial surface. Two phenotypes of overwintering Buxus microphylla 'Wintergreen' red leaves, with different measuring site and leaf thickness, were investigated in order to elucidate how red pigments in the outer leaf layer affected the Chl a fluorescence (F/F) and photochemical reflectance index. Our results showed that the F/F measured on leaves with the same red surface, but different leaf thickness, exhibited a slightly lower value in half leaf (separated upper and lower layers of leaves by removing the leaf edge similarly as affected by winter freezing and thawing) than that in the intact leaf (without removing the leaf edge), and the F/F measured on the red surface was significantly lower than that on the inner or backlighted green surface of the same thickness. Our results suggest that the usual measurement of F/F on red adaxial surface overestimates the actual degree of photoinhibition compared with that of the whole leaf in the winter. [ABSTRACT FROM AUTHOR]

* Background and Aims In temperate forests, some green plants, namely pyroloids (Pyroleae, Ericaceae) and some orchids, independently evolved a mode of nutrition mixing photosynthates and carbon gained from their mycorrhizal fungi (mixotrophy). Fungal carbon is more enriched in 13C than photosynthates, allowing estimation of the proportion of carbon acquired heterotrophically from fungi in plant biomass. Based on 13C enrichment, mixotrophic orchids have previously been shown to increase shoot autotrophy level over the growth season and with environmental light availability. But little is known about the plasticity of use of photosynthetic versus fungal carbon in pyroloids. * Methods Plasticity of mixotrophy with leaf age or light level (estimated from canopy openness) was investigated in pyroloids from three Estonian boreal forests. Bulk leaf 13C enrichment of five pyroloid species was compared with that of control autotrophic plants along temporal series (over one growth season) and environmental light gradients (n=405 samples). * Key Results Mixotrophic 13C enrichment was detected at studied sites for Pyrola chlorantha and Orthilia secunda (except at one site for the latter), but not for Chimaphila umbellata, Pyrola rotundifolia and Moneses uniflora. Enrichment with 13C did not vary over the growth season or between leaves from current and previous years. Finally, although one co-occurring mixotrophic orchid showed 13C depletion with increasing light availability, as expected for mixotrophs, all pyroloids responded identically to autotrophic control plants along light gradients. * Conclusions A phylogenetic trend previously observed is further supported: mixotrophy is rarely supported by 13C enrichment in the Chimaphila + Moneses clade, whereas it is frequent in the Pyrola + Orthilia clade. Moreover, pyroloid mixotrophy does not respond plastically to ageing or to light level. This contrasts with the usual view of a convergent evolution with orchids, and casts doubt on the way pyroloids use the carbon gained from their mycorrhizal fungi, especially to replace photosynthetic carbon. [ABSTRACT FROM AUTHOR]

Wintergreen oil is obtained from the leaves of aGaultheria procumbensand is commonly used as a folk remedy. On the other hand neuroblastoma is a childhood cancer of the sympathetic nervous system that originates from developing neural crest cells and is the second most common solid tumour. It was aimed to investigate the biological activity of Wintergreen oil on primary rat healthy neuron and N2a neuroblastoma cell in relation with concentration for the first time in the extent of this study. The effects of Wintergreen oil at various concentrations (0-400 mg/L) on cell proliferation were determined on both cell culture types by 3-(4,5 dimetylthiazol -2-yl) - 2,5 diphenltetrazolium bromide assay. And total antioxidant capacity and total oxidative stress parameters were used for the assessments ofin vitrooxidative effects. In addition, the genetic effect of Wintergreen oil on healthy and cancerous brain cells was assessed by single cell gel electrophoresis (or Comet assay). Our results clearly revealed that Wintergreen oil exhibited antioxidant activities at their low concentrations and cytotoxic at high concentrations. The mean values of the total scores of cells showing DNA damage (for Comet assay) was not found significantly different from the control values in both cell types. Again, Wintergreen oil showed weak anticarcinogenic activity against neuroblastoma cells. In conclusion, within the light of the findings of thisin vitrostudy, it is possible to denote that Wintergreen oil has low potential in cancer prevention and therapy. [ABSTRACT FROM AUTHOR]

Context: The number of multidrug resistant (MDR) microorganisms is increasing and the antimicrobial resistance expressed by these pathogens is generating a rising global health crisis. In fact, there are only a few antimicrobial agents left that can be used against MDR bacteria and fungi. Objective: In this study, the antimicrobial activities of selected natural products from the flora of Northern Ontario against selected microorganisms are reported. Materials and methods: Plants were collected from Sault Ste. Marie, Ontario, Canada, and ethanol extracts were prepared using EtOH:H2O (1:1, v/v). Fungal cultures used in this study were Candida albicans ATCC 10231 and Schizosaccharomyces octosporus. Bacterial cultures employed included Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Mycobacterium phlei ATCC 11758, and Streptococcus lactis ATCC 19435. The microplate resazurin assay was used to screen for antimicrobial activity. Results: Extracts of four plant species Chimaphila umbellata L. (Pyrolaceae), Betula papyrifera Marshall (Betulaceae), Rhus typhina L. (Anacardiaceae), and Fraxinus pennsylvanica Marshall (Oleaceae), and six compounds (gallic acid, ethyl gallate, caffeic acid, sinapic acid, gentisic acid, and chlorogenic acid) demonstrated antibacterial or antifungal activities with MICs ranging from 62.5 to 1000 µg/mL, respectively, for a chemical fraction of an extract from Betula papyrifera against the bacterium S. aureus. Discussion and conclusion: The present study has shown that certain plant extracts and select fractions and standard chemical compounds exhibit antimicrobial effects. Prince's Pine, Chimaphila umbellate, White Birch, Betula papyrifera, Staghorn Sumac, Rhus typhina, and Green Ash, Fraxinus pennsylvanica were the principal extracts exhibiting notable antibacterial and/or antifungal activities; while gallic acid, ethyl gallate, and caffeic acid demonstrated antibacterial activities and sinapic acid, gentisic acid, and chlorogenic acid demonstrated antifungal activities. [ABSTRACT FROM AUTHOR]

The rare forest plant Chimaphila umbellata (Ericaceae) has decreased drastically during the last century, approximately by 80 % in some regions in Sweden. We examined associations between various biotic and abiotic conditions related to changes in forest management and nitrogen deposition, and C. umbellata population size, flowering frequency, fruit set and seed production. Environmental conditions at 38 C. umbellata sites in the provinces of Uppland and Södermanland, Sweden, included light inflow, cover of competitive species, soil nitrogen, continuity of forest cover and soil texture. The results suggested that population size was negatively affected by cover of competitive species. Population size was not related to light availability although increased shading was associated with decreased flowering frequency. Fruit set was negatively affected by cover of competitive species, and seed production decreased with increasing soil nitrogen content. Fruit set and seed production increased with increasing population size. This study shows that denser forest stands and increased abundance of Vaccinium myrtillus and graminoid species may have strong negative effects on C. umbellata. This species' longevity and clonal propagation may buffer some negative impacts, which in turn might contribute to an extinction debt. To maintain viable populations of Chimaphila umbellata in Swedish forests, there is a need for targeted management in forest habitats, i.e. reintroducing moderate disturbance regimes to reduce competition and increase light inflow. [ABSTRACT FROM AUTHOR]

Dust seeds are the smallest seeds in angiosperms weighing just about a few micrograms. These seeds are characteristic of most orchids, and several studies have been performed on seed features, fecundity and dispersal of orchid dust seeds. In this study we examine seed features, seed production and seed dispersal in another plant group with dust seeds, the Pyroleae (Monotropoideae, Ericaceae), focusing on six species: Pyrola chlorantha, P. minor, P. rotundifolia, Chimaphila umbellata, Moneses uniflora and Orthilia secunda. Seed production per capsule among these species was in the range between ca 1000 and 7800, and seed production per capsule bearing shoot was in the range between ca 7000 and 60 000. Combining our results with published information on pollen-ovule ratios suggests that these Pyroleae species have a generally efficient pollination system. The most fecund species was P. minor, the only species among the investigated that is probably largely self-pollinating. The investigated Pyroleae species have a seed production comparable to the less fecund orchid species. We studied seed dispersal in the field in one of the species, P. chlorantha. Despite the extremely small and potentially buoyant seeds, the vast majority of seeds are deposited close to the seed source, within a few meters. Further studies on the recruitment ecology of the investigated Pyroleae species are currently under way. [ABSTRACT FROM AUTHOR]

The perennial evergreen herb, Chimaphila japonica is found exclusively in East Asian temperate coniferous or sometimes in deciduous forests. By using the Fast Isolation by Amplified Fragment Length Polymorphism (AFLP) of Sequences Containing repeats (FIASCO) protocol, 20 microsatellite primer sets were identified in two wild populations. Of these primers, 16 displayed polymorphisms and 4 were monomorphic. The number of alleles per locus ranged from one to six among populations, values for expected and observed heterozygosities ranged from 0.000 to 0.848 and from 0.000 to 1.000, respectively. The new SSR markers will be useful in obtaining estimates of population-level genetic diversity and in phylogeographic studies of C. japonica. [ABSTRACT FROM AUTHOR]

Genera in the tribe Pyroleae (subfamily Monotropoideae, family Ericaceae) occur as understory plants in northern temperate zones where some form major components of ecosystems. Most have been poorly studied in terms of their association with symbiotic fungi. In this study, colonization patterns of mycorrhizal roots of five members of the Pyroleae (Pyrola asarifolia Michx., Pyrola chlorantha Sw., Orthilia secunda (L.) House, Chimaphila umbellata (L.) W. Bart., Moneses uniflora (L.) Gray) were explored. Root samples were processed for light, fluorescence, and laser scanning confocal, scanning electron, and transmission electron microscopy, as well as for immunocytochemistry. Roots of all species had enlarged epidermal cells containing hyphal complexes, Hartig nets confined to the epidermis, and mantles. Epidermal cells were penetrated by hyphae originating from the Hartig net at more than one site either along the inner tangential wall or radial walls. The outer tangential wall of epidermal cells of all species, except M. uniflora, was thicker than radial and inner tangential walls and consisted of two layers, the outer containing nonesterified pectins that were labeled with JIM 5 antibodies. Radial walls and inner tangential walls did not label, but cortical cell walls did. Intracellular hyphal complexes developed initially around centrally positioned, enlarged epidermal cell nuclei and, through branching, occupied most of the cell volume. Senescence and degradation of the complexes followed. The fungal species in these symbiotic associations may be important functionally in nutrient exchange, as well as in contributing to broader linkages with other hosts in these plant communities. [ABSTRACT FROM AUTHOR]

Abstract: Bioassay-guided fractionation of Chimaphila umbellata (L.) W. Bart (Pyrolaceae) ethanol extracts led to the identification of 2,7-dimethyl-1,4-naphthoquinone (chimaphilin) as the principal antifungal component. The structure of chimaphilin was confirmed by 1H and 13C NMR spectroscopy. The antifungal activity of chimaphilin was evaluated using the microdilution method with Saccharomyces cerevisiae (0.05mg/mL) and the dandruff-associated fungi Malassezia globosa (0.39mg/mL) and Malassezia restricta (0.55mg/mL). Pronounced antioxidant activity of C. umbellata crude extract was also identified using the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay, suggesting this phytomedicine has an antioxidant function in wound healing. A chemical-genetic profile was completed with chimaphilin using ∼4700 S. cerevisiae gene deletion mutants. Cellular roles of deleted genes in the most susceptible mutants and secondary assays indicate that the targets for chimaphilin include pathways involved in cell wall biogenesis and transcription. [Copyright &y& Elsevier]

Lectotypes are designated for four of the six Linnaean names in Pyrola. The names are applied now to species of Chimaphila, Moneses, Orthilia, and Pyrola (Ericaceae, Pyroloideae). [ABSTRACT FROM AUTHOR]