Synergistic Inhibiting Effect of Phytochemicals in Rheum palmatum on Tyrosinase Based on Metabolomics and Isobologram Analyses.

Tyrosinase (TYR) plays a key role in the enzymatic reaction that is responsible for a range of unwanted discoloration effects, such as food browning and skin hyperpigmentation. TYR inhibitors could, therefore, be candidates for skin care products that aim to repair pigmentation problems. In this study, we used a metabolomics approach combined with the isobologram analysis to identify anti-TYR compounds within natural resources, and evaluate their possible synergism with each other. Rheum palmatum was determined to be a model plant for observing the effect, of which seven extracts with diverse phytochemicals were prepared by way of pressurized solvent extraction. Each Rheum palmatum extract (RPE) was profiled using nuclear magnetic resonance spectroscopy and its activity of tyrosinase inhibition was evaluated. According to the orthogonal partial least square analysis used to correlate phytochemicals in RPE with the corresponding activity, the goodness of fit of the model (R2 = 0.838) and its predictive ability (Q2 = 0.711) were high. Gallic acid and catechin were identified as the active compounds most relevant to the anti-TYR effect of RPE. Subsequently, the activity of gallic acid and catechin were evaluated individually, and when combined in various ratios by using isobologram analysis. The results showed that gallic acid and catechin in the molar ratios of 9:5 and 9:1 exhibited a synergistic inhibition on TYR, with a combination index lower than 0.77, suggesting that certain combinations of these compounds may prove effective for use in cosmetic, pharmaceutical, and food industries.

International policies and challenges on the legalization of traditional medicine/herbal medicines in the fight against COVID-19.

The coronavirus disease 2019 (COVID-19) has now rapidly spread around the world, causing an outbreak of acute infectious pneumonia. To develop effective and safe therapies for the prevention and treatment of COVID-19 has become the major global public health concern. Traditional medicine (TM)/herbal medicines (HMs) have been used to treat multiple epidemics in human history, which brings hope for the fight against COVID-19 in some areas. For example, in China, India, and South Korea with traditional medication history and theory, the governments issued a series of guidelines to support TM/HMs in the medication of COVID-19. In contrast, other countries e.g. North American and European governments are typically silent on these practices, unless to warn of possible harm and overselling. Such difference is due to the discrepancy in culture, history and philosophical views of health care and medication, as well as unharmonized policies and standards in the regulation and legalization of TM/HMs among different areas. Herein, we reviewed the responses and scientific researches from seven selected countries on the policies and legalization of TM/HMs to treat COVID-19, and also analyzed the major challenges and concerns to utilize the traditional knowledge and resource.

Modelling the dynamics of polar auxin transport in inflorescence stems of Arabidopsis thaliana.

The polar transport of the plant hormone auxin has been the subject of many studies, several involving mathematical modelling. Unfortunately, most of these models have not been experimentally verified. Here we present experimental measurements of long-distance polar auxin transport (PAT) in segments of inflorescence stems of Arabidopsis thaliana together with a descriptive mathematical model that was developed from these data. It is based on a general advection-diffusion equation for auxin density, as suggested by the chemiosmotic theory, but is extended to incorporate both immobilization of auxin and exchange with the surrounding tissue of cells involved in PAT, in order to account for crucial observations. We found that development of the present model assisted effectively in the analysis of experimental observations. As an example, we discuss the analysis of a quadruple mutant for all four AUX1/LAX1-LAX3 influx carriers genes. We found a drastic change in the parameters governing the exchange of PAT channels with the surrounding tissue, whereas the velocity was still of the order of magnitude of the wild type. In addition, the steady-state flux of auxin through the PAT system of the mutant did not exhibit a saturable component, as we found for the wild type, suggesting that the import carriers are responsible for the saturable component in the wild type. In the accompanying Supplementary data available at JXB online, we describe in more detail the data-driven development of the model, review and derive predictions from a mathematical model of the chemiosmotic theory, and explore relationships between parameters in our model and processes and parameters at the cellular level.

TNFα secretion of monocytes exposed to pulsed radiofrequency treatment: a possible working mechanism of PRF chronic pain management.

Pulsed radiofrequency treatment (PRF) is a promising new technique increasingly used in treatment of chronic pain. The molecular working mechanism of PRF is not exactly known and is currently being investigated. This study investigates a possible role of PRF-induced modulation of TNFα secretion by differentiated monocytes in chronic pain management. The results show no significant PRF-induced change in TNFα secretion of lipopolysaccharides (LPS)-stimulated monocytes. However, PRF does significantly increase TNFα secretion of differentiated monocytes that have not been stimulated with LPS. This may indicate a possible role of PRF treatment in increasing TNFα production of nonstimulated monocytes. More research is needed to determine whether this is truly a part of the working mechanism of PRF in chronic pain management and which other factors are involved.

Valorization of sorghum ash with digestate and biopreparations in the development biomass of plants in a closed production system of energy.

Replacing chemical fertilizers with non-toxic waste that meet all fertilizing purposes, including ash from plant biomass and their management is becoming the important goal of sustainable agriculture concerning energy plants production in a closed system. This study aims to explore a novel strategy for utilizing natural sorghum ash together with digestate and ecological compounds, to replace synthetic fertilizers, for the energy plant development improvement and thus reduction of the environment pollution. Sorghum, as an energy plant, cultivated in low quality sandy and podzolic soils, in Central and North Poland climate, was fertilized with different doses of YaraMila Complex, a synthetic fertilizer (0, 150, 300 kg ha-1 Each dose was supplemented with different amounts of sorghum ash (0.5, 1, 2 and 4 t ha-1), used alone or with addition of APOL-HUMUS (soil improver; 10 L ha-1), biogas plant digestate (30 m3 ha-1) and Stymjod (nano-organic leaf fertilizer; 5 L ha-1). Added to each YaraMila Complex dose, the applied ash amounts (optimally 2-4 t ha-1), increased growth of plants, crop biomass, index of chlorophyll content, net photosynthesis, transpiration, stomatal conductance, content of intercellular CO2, activity of acid and alkaline phosphatase, RNase and dehydrogenase and energy properties. Sorghum ash used with the lesser YaraMila Complex doses of 0 or 150 kg ha-1 caused the enhanced growth of plants more than the doubled YaraMila Complex amounts applied alone (150 or 300 kg ha-1, correspondingly). Additionally, applied biogas plant digestate, APOL-HUMUS and Stymjod further increased the plant growth. This indicates that the application of natural sorghum ash accelerates energy plant development, can reduce by half the recommended synthetic fertilizer doses on poor and marginal soil and enables the cultivation of sorghum in a closed production cycle.

The Anti-Inflammatory Action of Pulsed Radiofrequency-A Hypothesis and Potential Applications.

In 2013, it was reported that pulsed radiofrequency (PRF) could be applied to obtain a systemic anti-inflammatory effect. Patients with chronic pain and patients with an inflammatory condition from other disciplines could potentially profit from this finding. At that time, intravenous application was used, but since then, it became clear that it could be applied transcutaneously as well. This procedure was named RedoxPRF. This can be used both for regional and for systemic application. Recently, the basic element of the mode of action has been clarified from the analysis of the effects of PRF on a standard model of muscle injury in rats. The objective of this paper is to present a hypothesis on the mode of action of RedoxPRF now that the basic mechanism has become known. Cell stress causes an increased production of free radicals, disturbing the redox equilibrium, causing oxidative stress (OS) either directly or secondarily by other types of stress. Eventually, OS causes inflammation and an increased sympathetic (nervous) system activity. In the acute form, this leads to immune paralysis; in the chronic form, to immune tolerance and chronic inflammation. It is hypothesized that RedoxPRF causes a reduction of free radicals by a recombination of radical pairs. For systemic application, the target cells are the intravascular immune cells that pass through an activated area as on an assembly line. Hypothesis conclusions: 1. RedoxPRF treatment works selectively on OS. It has the unique position of having a point of engagement at the most upstream level of the train of events. 2. RedoxPRF has the potential of being a useful tool in the treatment of inflammatory diseases and possibly of stage 4 cancer. 3. In the treatment of chronic pain, RedoxPRF is an entirely new method because it is different from ablation as well as from stimulation. We propose the term "functional restoration". 4. Controlled studies must be conducted to develop this promising new field in medicine further.

3'sialyllactose and 6'sialyllactose enhance performance in endurance-type exercise through metabolic adaptation.

Human milk oligosaccharides (HMOs) belong to a group of multifunctional glycans that are abundantly present in human breast milk. While health effects of neutral oligosaccharides have been investigated extensively, a lot remains unknown regarding health effects of acidic oligosaccharides, such as the two sialyllactoses (SLs), 3'sialyllactose (3'SL), and 6'sialyllactose (6'SL). We utilized Caenorhabditis elegans (C. elegans) to investigate the effects of SLs on exercise performance. Using swimming as an endurance-type exercise, we found that SLs decrease exhaustion, signifying an increase in endurance that is strongest for 6'SL. Through an unbiased metabolomics approach, we identified changes in energy metabolism that correlated with endurance performance. Further investigation suggested that these metabolic changes were related to adaptations of muscle mitochondria that facilitated a shift from beta oxidation to glycogenolysis during exercise. We found that the effect of SLs on endurance performance required AMPK- (aak-1/aak-2) and adenosine receptor (ador-1) signaling. We propose a model where SLs alter the metabolic status in the gut, causing a signal from the intestine to the nervous system toward muscle cells, where metabolic adaptation increases exercise performance. Together, our results underline the potential of SLs in exercise-associated health and contribute to our understanding of the molecular processes involved in nutritionally-induced health benefits.

Polar auxin transport: an early invention.

In higher plants, cell-to-cell polar auxin transport (PAT) of the phytohormone auxin, indole-3-acetic acid (IAA), generates maxima and minima that direct growth and development. Although IAA is present in all plant phyla, PAT has only been detected in land plants, the earliest being the Bryophytes. Charophyta, a group of freshwater green algae, are among the first multicellular algae with a land plant-like phenotype and are ancestors to land plants. IAA has been detected in members of Charophyta, but its developmental role and the occurrence of PAT are unknown. We show that naphthylphthalamic acid (NPA)-sensitive PAT occurs in internodal cells of Chara corallina. The relatively high velocity (at least 4-5 cm/h) of auxin transport through the giant (3-5 cm) Chara cells does not occur by simple diffusion and is not sensitive to a specific cytoplasmic streaming inhibitor. The results demonstrate that PAT evolved early in multicellular plant life. The giant Chara cells provide a unique new model system to study PAT, as Chara allows the combining of real-time measurements and mathematical modelling with molecular, developmental, cellular, and electrophysiological studies.

Extracting relevant physiological information from polar auxin transport data in Panax ginseng.

BACKGROUND: Measuring polar auxin transport (PAT) in plants and drawing conclusions from the observed transport data is only meaningful if these data are being analysed with a mathematical model which describes PAT. In this report we studied the polar auxin transport in Panax ginseng stems of different age and grown on different substrates. METHODS: We measured polar IAA transport in stems using a radio labelled IAA and analysed the transport data with a mathematical model we developed for Arabidopsis. RESULTS: We found that PAT in ginseng stems, as compared to Arabidopsis inflorescence stems, has a 2-fold lower transport velocity and a 3-fold lower steady state auxin flux. CONCLUSION: We were able to pinpoint two physiological parameters that influenced the observed transport characteristics in ginseng which differ from Arabidopsis, namely an increase in immobilization together with a reduced reflux of IAA from the surrounding tissue back to the transporting cells.

Evolutionary and Functional Analysis of a Chara Plasma Membrane H+-ATPase.

H+-ATPases are the main transporters in plant and fungal plasma membranes (PMs), comparable to the Na+/K+ ATPases in animal cells. At the molecular level, most studies on the PM H+-ATPases have been focused on land plants and fungi (yeast). The research of PM H+-ATPases in green algae falls far behind due to the lack of genetic information. Here we studied a potential PM H+-ATPase (CHA1) from Chara australis, a species of green algae belonging to the division Charophyta, members of which are considered to be one of the closest ancestors of land plants. The gene encodes a 107 kDa protein with all 6 P-type ATPase-specific motifs and a long, diverse C-terminal domain. A new amino acid sequence motif R*****Q in transmembrane segment 5 was identified among the known PM H+-ATPases from Charophyta and Chlorophyta algae, which is different from the typical PM H+-ATPases in yeast or land plants. Complementation analysis in yeast showed that CHA1 could successfully reach the PM, and that proton pump activity was obtained when the last 77 up to 87 amino acids of the C-terminal domain were deleted. PM localization was confirmed in Arabidopsis protoplasts; however, deletion of more than 55 amino acids at the N-terminus or more than 98 amino acids at the C-terminus resulted in failure of CHA1 to reach the PM in yeast. These results suggest that an auto-inhibition domain is located in the C-terminal domain, and that CHA1 is likely to have a different regulation mechanism compared to the yeast and land plant PM H+-ATPases.

Steamed Panax notoginseng Attenuates Anemia in Mice With Blood Deficiency Syndrome via Regulating Hematopoietic Factors and JAK-STAT Pathway.

Panax notoginseng (Burk.) F. H. Chen is a medicinal herb used to treat blood disorders since ancient times, of which the steamed form exhibits the anti-anemia effect and acts with a "blood-tonifying" function according to traditional use. The present study aimed to investigate the anti-anemia effect and underlying mechanism of steamed P. notoginseng (SPN) on mice with blood deficiency syndrome induced by chemotherapy. Blood deficiency syndrome was induced in mice by cyclophosphamide and acetylphenylhydrazine. A number of peripheral blood cells and organs (liver, kidney, and spleen) coefficients were measured. The mRNA expression of hematopoietic function-related cytokines in the bone marrow of mice was detected by RT-qPCR. The janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway was screened based on our previous analysis by network pharmacology. The expression of related proteins and cell cycle factors predicted in the pathway was determined by Western blot and RT-qPCR. SPN could significantly increase the numbers of peripheral blood cells and reverse the enlargement of spleen in a dose-dependent manner. The quantities of related hematopoietic factors in bone marrow were also increased significantly after SPN administration. SPN was involved in the cell cycle reaction and activation of immune cells through the JAK-STAT pathway, which could promote the hematopoiesis.

Auxin effects on ion transport in Chara corallina.

The plant hormone auxin has been widely studied with regard to synthesis, transport, signaling and functions among the land plants while there is still a lack of knowledge about the possible role for auxin regulation mechanisms in algae with "plant-like" structures. Here we use the alga Chara corallina as a model to study aspects of auxin signaling. In this respect we measured auxin on membrane potential changes and different ion fluxes (K(+), H(+)) through the plasma membrane. Results showed that auxin, mainly IAA, could hyperpolarize the membrane potential of C. corallina internodal cells. Ion flux measurements showed that the auxin-induced membrane potential change may be based on the change of K(+) permeability and/or channel activity rather than through the activation of proton pumps as known in land plants.

Cellular Auxin Transport in Algae.

The phytohormone auxin is one of the main directors of plant growth and development. In higher plants, auxin is generated in apical plant parts and transported from cell-to-cell in a polar fashion. Auxin is present in all plant phyla, and the existence of polar auxin transport (PAT) is well established in land plants. Algae are a group of relatively simple, autotrophic, photosynthetic organisms that share many features with land plants. In particular, Charophyceae (a taxon of green algae) are closest ancestors of land plants. In the study of auxin function, transport and its evolution, the algae form an interesting research target. Recently, proof for polar auxin transport in Chara species was published and auxin related research in algae gained more attention. In this review we discuss auxin transport in algae with respect to land plants and suggest directions for future studies.

Activation of the oxidative burst by yeast elicitor in Catharanthus roseus cells occurs independently of the activation of genes involved in alkaloid biosynthesis.

In Catharanthus roseus cell suspensions, expression of several terpenoid indole alkaloid (TIA) biosynthetic genes, including those encoding strictosidine synthase and tryptophan decarboxylase, is coordinately induced by fungal elicitors such as yeast extract (YE). This induction is mediated by several signaling steps including the biosynthesis of jasmonic acid, and the activation of the jasmonic acid-responsive ORCA transcription factors. We investigated a possible role of reactive oxygen species (ROS) as a second messenger in this system. YE was shown to activate the production of ROS, which was dependent on protein phosphorylation and calcium influx. However, ROS generation was neither necessary for the induction of genes involved in TIA biosynthesis by YE nor by itself sufficient to induce these genes. Therefore, we conclude that activation of the oxidative burst by YE occurs independently of the activation of genes involved in TIA biosynthesis.

Zinc finger proteins act as transcriptional repressors of alkaloid biosynthesis genes in Catharanthus roseus.

In Catharanthus roseus cell suspensions, the expression of several terpenoid indole alkaloid biosynthetic genes, including two genes encoding strictosidine synthase (STR) and tryptophan decarboxylase (TDC), is coordinately induced by fungal elicitors such as yeast extract. To identify molecular mechanisms regulating the expression of these genes, a yeast one-hybrid screening was performed with an elicitor-responsive part of the TDC promoter. This screening identified three members of the Cys(2)/His(2)-type (transcription factor IIIA-type) zinc finger protein family from C. roseus, ZCT1, ZCT2, and ZCT3. These proteins bind in a sequence-specific manner to the TDC and STR promoters in vitro and repress the activity of these promoters in trans-activation assays. In addition, the ZCT proteins can repress the activating activity of APETALA2/ethylene response-factor domain transcription factors, the ORCAs, on the STR promoter. The expression of the ZCT genes is rapidly induced by yeast extract and methyljasmonate. These results suggest that the ZCT proteins act as repressors in the regulation of elicitor-induced secondary metabolism in C. roseus.