Folk medicine, biological activity, and chemical profiles of Brazilian Acanthaceae (Lamiales) - A review.

INTRODUCTION: The botanical family Acanthaceae (order Lamiales) potentially comprises 4900 species in 191 genera with extensive morphological, habit and habitat diversity. The family is widely distributed throughout the world but is especially rich in tropical and subtropical regions. Many of its species have great ornamental importance and are broadly used for medicinal purposes in several countries of Asia and Africa. Brazil is a main center of diversity of the family, where they are distributed across all its biomes, mainly in the herbaceous-shrub stratum. Medicinal investigations about Brazilian species are scarce, the exception being a single native species, Justicia pectoralis Jacq., that is widely used and studied chemically. AIM OF THE REVIEW: This work compiled studies that indicated folk medicinal use, investigated biological activity, or evaluated the chemical composition of Brazilian species of Acanthaceae. MATERIAL AND METHODS: Medicinal uses, investigations of biological activities and chemical data were collected and summarized through bibliographic surveys. Tables were compiled to standardize the information and the appropriate references were gathered for each species. Registration of chemical components used in the treatment of ailments and in preserving health were emphasized with the aim of stimulating future investigations. RESULTS: The breadths of habitats and morphologies of the family are directly related to its chemical diversity, as confirmed here for Brazilian species. Although the investigated species represent less than 9% of the total richness of the family in Brazil, they encompass a great diversity of chemical substances. The data indicated folk medicinal uses for 26 species and biological tests for 23, while 30 species were investigated chemically. Ruellia and Justicia were the most researched genera with 12 and 11 species, representing approximately 14% and 7% of Brazilian species of each genus, respectively. Two species are native to other countries but become naturalized in Brazil. Studies of native species were carried out in different countries around the world, with many reports of medicinal uses and biological tests. Examples of uses include anticancer and antidepressant actions, as well as activities against respiratory problems and other diseases. CONCLUSIONS: This work highlights the chemical and biological diversity of the studied Brazilian species of Acanthaceae, which emphasizes the need to expand studies with native Brazilian species.

Perspectives on antimicrobial properties of Paulownia tomentosa Steud. fruit products in the control of Staphylococcus aureus infections.

ETHNOPHARMACOLOGICAL RELEVANCE: Paulownia tomentosa Steud. (P. tomentosa) is a medium-sized tree traditionally used in Chinese folk medicine for the treatment of infectious diseases. It is a rich source of prenylated phenolic compounds that have been extensively studied for their promising biological activities. AIM OF THE STUDY: Due to the increasing development of antibiotic resistance, our study investigated plant-derived natural products from the fruits of P. tomentosa that could control Staphylococcus aureus infections with novel targets/modes of action and reduce antimicrobial resistance. MATERIALS AND METHODS: The ethanolic extract was fractionated and detected by liquid chromatography. The antistaphylococcal effects of the plant formulations were studied in detail in vitro by various biological methods, including microdilution methods for minimum inhibitory concentration (MIC), the checkerboard titration technique for synergy assay, fluorescence measurements for membrane disruption experiments, autoinducer-2-mediated bioassay for quorum sensing inhibition, and counting of colony-forming units for relative adhesion. Morphology was examined by transmission electron microscopy. RESULTS: Total ethanolic extract and chloroform fraction showed MICs of 128 and 32 µg/mL, respectively. Diplacol, diplacone, and 3'-O-methyl-5'-hydroxydiplacone inhibited S. aureus growth in the range of 8-16 µg/mL. Synergistic potential was shown in combination with mupirocin and fusidic acid. The ethanolic extract and the chloroform fraction destroyed the cell membranes by 91.61% and 79.46%, respectively, while the pure compounds were less active. The ethanolic extract and the pure compounds reduced the number of adhered cells to 47.33-10.26% compared to the untreated control. All tested plant formulations, except diplacone, inhibited quorum sensing of S. aureus. Transmission electron microscopy showed deformation of S. aureus cells. CONCLUSIONS: The products from the fruit of P. tomentosa showed antimicrobial properties against S. aureus alone and in combination with antibiotics. By affecting intracellular targets, geranylated flavonoids proposed novel approaches in the control of staphylococcal infections.

Haberlea rhodopensis Extract Tunes the Cellular Response to Stress by Modulating DNA Damage, Redox Components, and Gene Expression.

Ionizing radiation (IR) and reactive oxygen species (ROS)-induced oxidative stress can cause damage to cellular biomolecules, including DNA, proteins, and lipids. These harmful effects can compromise essential cellular functions and significantly raise the risk of metabolic dysfunction, accumulation of harmful mutations, genome instability, cancer, accelerated cellular senescence, and even death. Here, we present an investigation of HeLa cancer cells' early response to gamma IR (γ-IR) and oxidative stress after preincubation of the cells with natural extracts of the resurrection plant Haberlea rhodopensis. In light of the superior protection offered by plant extracts against radiation and oxidative stress, we investigated the cellular defence mechanisms involved in such protection. Specifically, we sought to evaluate the molecular effects of H. rhodopensis extract (HRE) on cells subjected to genotoxic stress by examining the components of the redox pathway and quantifying the transcription levels of several critical genes associated with DNA repair, cell cycle regulation, and apoptosis. The influence of HRE on genome integrity and the cell cycle was also studied via comet assay and flow cytometry. Our findings demonstrate that HREs can effectively modulate the cellular response to genotoxic and oxidative stress within the first two hours following exposure, thereby reducing the severity of such stress. Furthermore, we observed the specificity of genoprotective HRE doses depending on the source of the applied genotoxic stress.

Chromosome-scale assembly of the Verbenaceae species Queen's Wreath (Petrea volubilis L.).

OBJECTIVES: Petrea volubilis, a member of the Order Lamiales and the Verbenaceae family, is an important horticultural species that has been used in traditional folk medicine. To provide a genome sequence for comparative studies within the Order Lamiales that includes important families such as Lamiaceae (mints), we generated a long-read, chromosome-scale genome assembly of this species. DATA DESCRIPTION: Using a total of 45.5 Gb of Pacific Biosciences long read sequence, we generated a 480.2 Mb assembly of P. volubilis, of which, 93% is chromosome anchored. Representation of genic regions was robust with 96.6% of the Benchmarking of Universal Single Copy Orthologs present in the genome assembly. A total of 57.8% of the genome was annotated as a repetitive sequence. Using a gene annotation pipeline that included refinement of gene models using transcript evidence, 30,982 high confidence genes were annotated. Access to the P. volubilis genome will facilitate evolutionary studies in the Lamiales, a key order of Asterids that includes significant crop and medicinal plant species.

[Complete chloroplast genome sequencing and phylogeny of wild Atractylodes lancea from Yuexi, Anhui province].

This study investigated the choroplast genome sequence of wild Atractylodes lancea from Yuexi in Anhui province by high-throughput sequencing, followed by characterization of the genome structure, which laid a foundation for the species identification, analysis of genetic diversity, and resource conservation of A. lancea. To be specific, the total genomic DNA was extracted from the leaves of A. lancea with the improved CTAB method. The chloroplast genome of A. lancea was sequenced by the high-throughput sequencing technology, followed by assembling by metaSPAdes and annotation by CPGAVAS2. Bioiformatics methods were employed for the analysis of simple sequence repeats(SSRs), inverted repeat(IR) border, codon bias, and phylogeny. The results showed that the whole chloroplast genome of A. lancea was 153 178 bp, with an 84 226 bp large single copy(LSC) and a 18 658 bp small single copy(SSC) separated by a pair of IRs(25 147 bp). The genome had the GC content of 37.7% and 124 genes: 87 protein-coding genes, 8 rRNA genes, and 29 tRNA genes. It had 26 287 codons and encoded 20 amino acids. Phylogenetic analysis showed that Atractylodes species clustered into one clade and that A. lancea had close genetic relationship with A. koreana. This study established a method for sequencing the chloroplast genome of A. lancea and enriched the genetic resources of Compositae. The findings are expected to lay a foundation for species identification, analysis of genetic diversity, and resource conservation of A. lancea.

Three new naphthoquinones from the tubers of Sinningia mauroana.

Three new naphthoquinones, 5,6,7-trimethoxydunnione (1), 6,7-dimethoxy-α-dunnione (2), and 5,6,7-trimethoxydunniol (3) were isolated from the tubers of Sinningia mauroana Chautems, together with eleven known compounds: 5-hydroxy-6,7-dimethoxy-α-dunnione (4), 6-hydroxy-7-methoxy-α-dunnione (5), 7-hydroxy-6-methoxy-α-dunnione (6), cedrol (7), tectoquinone (8), plantainoside A (9), calceolarioside A (10), calceolarioside B (11), sanangoside (12), allo-calceolarioside A (13), and conandroside (14). The known compounds 4, 7-9 and 11-13 are being reported for the first time in this species. Hexane, ethyl acetate and ethanol extracts were tested for antioxidant activity by the ORAC-FL method, and the antioxidant capacity was measured as trolox equivalent (TE). The ethyl acetate (8808 µg TE g-1) and ethanol (7911 µg TE g-1) extracts showed high antioxidant capacity, while the hexane extract displayed weak antioxidant capacity (860 µg TE g-1).

De Novo Hybrid Assembly of the Salvia miltiorrhiza Mitochondrial Genome Provides the First Evidence of the Multi-Chromosomal Mitochondrial DNA Structure of Salvia Species.

Salvia miltiorrhiza has been an economically important medicinal plant. Previously, an S. miltiorrhiza mitochondrial genome (mitogenome) assembled from Illumina short reads, appearing to be a single circular molecule, has been published. Based on the recent reports on the plant mitogenome structure, we suspected that this conformation does not accurately represent the complexity of the S. miltiorrhiza mitogenome. In the current study, we assembled the mitogenome of S. miltiorrhiza using the PacBio and Illumina sequencing technologies. The primary structure of the mitogenome contained two mitochondrial chromosomes (MC1 and MC2), which corresponded to two major conformations, namely, Mac1 and Mac2, respectively. Using two approaches, including (1) long reads mapping and (2) polymerase chain reaction amplification followed by Sanger sequencing, we observed nine repeats that can mediate recombination. We predicted 55 genes, including 33 mitochondrial protein-coding genes (PCGs), 3 rRNA genes, and 19 tRNA genes. Repeat analysis identified 112 microsatellite repeats and 3 long-tandem repeats. Phylogenetic analysis using the 26 shared PCGs resulted in a tree that was congruent with the phylogeny of Lamiales species in the APG IV system. The analysis of mitochondrial plastid DNA (MTPT) identified 16 MTPTs in the mitogenome. Moreover, the analysis of nucleotide substitution rates in Lamiales showed that the genes atp4, ccmB, ccmFc, and mttB might have been positively selected. The results lay the foundation for future studies on the evolution of the Salvia mitogenome and the molecular breeding of S. miltiorrhiza.

A review on ethnobotany, phytochemistry, and pharmacology of the genus Didymocarpus wall. (Gesneriaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants have been extensively used to treat various illnesses since the dawn of civilization. The genus Didymorcapus Wall. comprises 100 species widely distributed in the tropical regions of Asia, with a few found scattered in Africa and Australia. Species in this genus have long been used in folk medicine to treat various illnesses, including wounds, kidney stones, inflammations, asthma, flu, eczema, dysentery, fractures, colic etc. Some species have applications as weight loss agents, laxatives, and protective medication after childbirth. AIM: To provide comprehensive information on the current knowledge of the ethnobotanical uses, phytochemical compounds, pharmacological applications, and toxicology of genus Didymocarpus to reveal its therapeutic potential, offering insights into future research opportunities. MATERIALS AND METHODS: Data were systematically obtained from books and online databases such as PubMed, Web of Science, Scopus, Sci Finder, Google Scholar, Science direct, ACS Publications, Elsevier, Wiley Online Library. RESULTS: Seventeen Didymocarpus species have applications in traditional medicine in different Asian countries. A total of 166 compounds have been isolated from the genus Didymocarpus including terpenoids, flavonoids, phenolic compounds, fatty acids, chalcones, steroids, and others. Among these constituents, terpenoids, flavonoids, chalcones, and phenolics are the significant contributors to pharmacological activities of the genus Didymocarpus, possessing wide-reaching biological activities both in vivo and in vitro. The crude extracts and isolated phytochemical compounds from this genus have been shown to exhibit various pharmacological activities, including antiurolithiatic, nephro-protective, antimicrobial, anticancer, antidiabetic, cytotoxic, wound healing, and antioxidant activities. CONCLUSIONS: Traditional uses and scientific evaluation of Didymocarpus indicate that Didymocarpus pedicellata is one of the most widely used species in some parts of the world. Although substantial progress on the chemical and pharmacological properties of Didymocarpus species has been made, further studies on the pharmacology and toxicology of these species are needed to ensure safety, efficacy, and quality. Also, further research on the structure-activity relationship of some of the isolated phytocompounds may improve their biological potency and scientific exploitation of traditional uses of the Didymocarpus taxa.

C-geranylated flavonoids from Paulownia tomentosa Steud. fruit as potential anti-inflammatory agents.

ETHNOPHARMACOLOGICAL RELEVANCE: Paulownia tomentosa Steud., a traditional Chinese medicinal plant, was used for many centuries in Chinese herbal medicine as a component of remedies for many illnesses, including inflammatory diseases. It is a rich source of phenolic compounds, mainly geranylated flavonoids, which are currently studied for their promising biological activities. AIM OF THE STUDY: The study aimed to isolate minor geranylated flavanones and flavones from P. tomentosa fruit and evaluate their cytotoxicity and possible anti-inflammatory effects in a cell-based model of inflammation. MATERIALS AND METHODS: Chromatographic separation of chloroform portion of the ethanolic extract of P. tomentosa fruit led to the isolation of twenty-seven flavonoids (1-27), twenty-six of them geranylated with different modifications and one non-geranylated flavanone, and two phenolic compounds. Compounds were identified using UV, IR, HRMS, NMR, and CD spectroscopy. Ten of these compounds (7-10, 12, 21, 22, 24, 25, and 27) were determined to be new flavonoid derivatives obtained from a natural source for the first time. Selected compounds were analyzed for cytotoxicity and anti-inflammatory potential to affect the activation of nuclear factor κB/activator protein 1 (NF-κB/AP-1) after lipopolysaccharide (LPS) stimulation. RESULTS: All the test compounds (1-21 and 23-26) reduced the activation of NF-κB/AP-1 24 h after the addition of LPS. Eight compounds (5, 14-18, 21, and 26) were more active than prednisone, a widely used anti-inflammatory drug. However, this effect was not seen significantly on the level of TNF-α and IL-1ß, which can be explained by the plurality of possible outcomes of activation of the NF-κB pathway in cells. CONCLUSIONS: Results of the presented study confirmed that constituents from traditional Chinese medicinal plant P. tomentosa Steud. have promising anti-inflammatory activities and can serve as a potential source of inspiration for new anti-inflammatory medications.

Sequence Analysis of the Complete Mitochondrial Genome of a Medicinal Plant, Vitex rotundifolia Linnaeus f. (Lamiales: Lamiaceae).

In the present study, we depicted the complete mitochondrial genome of a valuable medicinal plant, Vitex rotundifolia. The mitochondrial genome of V. rotundifolia, mapped as a circular molecule, spanned 380,980 bp in length and had a GC content of 45.54%. The complete genome contained 38 protein-coding genes, 19 transfer RNAs (tRNAs), and 3 ribosomal RNAs (rRNAs). We found that there were only 38.73% (147.54 kb), 36.28% (138.23 kb), and 52.22% (198.96 kb) of the homologous sequences in the mitochondrial genome of V. rotundifolia, as compared with the mitochondrial genomes of Scutellaria tsinyunensis, Boea hygrometrica, and Erythranthe lutea, respectively. A multipartite structure mediated by the homologous recombinations of the three direct repeats was found in the V. rotundifolia mitochondrial genome. The phylogenetic tree was built based on 10 species of Lamiales, using the maximum likelihood method. Moreover, this phylogenetic analysis is the first to present the evolutionary relationship of V. rotundifolia with the other species in Lamiales, based on the complete mitochondrial genome.

Qualitative and Quantitative Analysis of Secondary Metabolites in Morphological Parts of Paulownia Clon In Vitro 112® and Their Anticoagulant Properties in Whole Human Blood.

It is not easy to find data in the scientific literature on the quantitative content of individual phytochemicals. It is possible to find groups of compounds and even individual compounds rather easily, but it is not known what their concentration is in cultivated or wild plants. Therefore, the subject of this study was to determine the content of individual compounds in the new Paulownia species, Oxytree, developed in a biotechnology laboratory in 2008 at La Mancha University in Spain. Six secondary metabolites were isolated, and their chemical structure was confirmed by spectral methods. An analytical method was developed, which was then used to determine the content of individual compounds in leaves, twigs, flowers and fruits of Paulownia Clon in Vitro 112®. No flavonoids were found in twigs and fruits of Oxytree, while the highest phenylethanoid glycosides were found in twigs. In this study, we also focused on biological properties (anticoagulant or procoagulant) of extract and four fractions (A-D) of different chemical composition from Paulownia Clon in Vitro 112 leaves using whole human blood. These properties were determined based on the thrombus-formation analysis system (T-TAS), which imitates in vivo conditions to assess whole blood thrombogenecity. We observed that three fractions (A, C and D) from leaves decrease AUC10 measured by T-TAS. In addition, fraction D rich in triterpenoids showed the strongest anticoagulant activity. However, in order to clarify the exact mechanism of action of the active substances present in this plant, studies closer to physiological conditions, i.e., in vivo studies, should be performed, which will also allow to determine the effects of their long-term effects.

Optimization of an ultrasonic-assisted aqueous two-phase extraction method for four flavonoids from Lysionotus pauciflorus.

An effective approach to extracting four flavonoids (lysioside C, nevadensin-7-sambubioside, ikarisoside B, and nevadensin) from Lysionotus pauciflorus using ultrasonic-assisted aqueous two-phase extraction (UAATPE) technology was studied. Ethanol/K2HPO4 system was selected and the influence of several important parameters, including composition of the aqueous two-phase system (ATPS), extraction temperature and time, particle size, and solvent to material ratio, were investigated by single factor experimentss. Then three key parameters (ethanol concentration, solvent to material ratio and extraction temperature) were further optimized by response surface methodology (RSM). The optimal process was that 1 g 120 mesh herb powders were extracted with 45 g ATPS (made of 30% ethanol/18% K2HPO4) in 43 °C for 30 min, and yields of four flavonoids could reach 2.56, 2.06, 3.62, and 6.28 mg/g, respectively. Compared with ultrasonic-assisted extraction (UAE) in three conventional solvents (water, ethanol and 60% ethanol), the UAATPE displayed comparatively higher extraction capability. Therefore, UAATPE in ethanol/K2HPO4 system could be an alternative technology for integrated extraction of the flavonoids from L. pauciflorus.

Valorization of Artichoke Industrial By-Products Using Green Extraction Technologies: Formulation of Hydrogels in Combination with Paulownia Extracts.

The integral valorization of artichoke bracts generated during industrial canning of artichoke was assessed. The extraction of bioactive compounds was addressed with pressurized hot water under subcritical conditions. The performance of this stage on the extraction of phenolics with antioxidant properties and the saccharidic fraction using conventional and microwave heating was compared. The microwave assisted process was more efficient than the conventional one regarding extraction yields of total solubles, and glucose and fructose oligomers and phenolics, because lower operational temperatures and shorter times were needed. Degradation of fructose oligomers was observed at temperatures higher than 160 °C, whereas the maximal phenolic content occurred at 220 °C. Both the extracts and the residual solids, obtained at conditions leading to maximum phenolics yields, were evaluated for the production of starch-based hydrogels, supplemented with Paulownia leaves' aqueous extracts.

Anti-hepatitis B virus activity and hepatoprotective effect of des(rhamnosyl) verbascoside from Lindernia ruellioides in vitro.

Although clinically approved hepatitis B virus (HBV) polymerase inhibitors (lamivudine-3TC, entecavir, etc.) serve as effective therapeutics, the virus can easily generate resistance to them. Therefore, the treatment of HBV infection remains a public health problem. Numerous studies have shown that natural products have prospective anti-HBV activity. The purpose of this study was to isolate and extract des(rhamnosyl) verbascoside from Lindernia ruellioides (Colsm.) Pennell and explore its anti-HBV and hepatoprotective effects. Anti-HBV activity was evaluated in HepG2.2.15 cells, a human hepatocellular carcinoma cell line with HBV-stable infection, and its protective effect was evaluated in HL-7702 cells, a normal human liver cell line. HepG2.2.15 cells maintained normal growth morphology within the selected concentration range of des(rhamnosyl) verbascoside. It also inhibited the expression of HBV antigens and HBV DNA in a dose- and time-dependent manner in vitro. Further, western blot experiments showed that it could downregulate HBV X protein (HBx) expression in a dose-dependent manner. In the H2 O2 -induced hepatocyte injury model, the cell-survival rate of the HL-7702 cells with the highest drug dose reached 85.25%, which was significantly improved compared with that of the model group. Most of the cells returned to normal morphology, showing polygonal or fusiform structures. Thus, it may be stated that des(rhamnosyl) verbascoside exhibits anti-HBV activity and hepatoprotective effects in vitro and may exert an anti-HBV effect via antigen inhibition, HBV DNA secretion, and HBx protein expression.

Flavone-Rich Fractions and Extracts from Oroxylum indicum and Their Antibacterial Activities against Clinically Isolated Zoonotic Bacteria and Free Radical Scavenging Effects.

Oroxylum indicum extracts from the seeds collected from Lampang and Pattani provinces in Thailand, and young fruits and flowers exhibited in vitro display antioxidant and antibacterial activities against clinically isolated zoonotic bacteria including Staphylococcus intermedius, Streptococcus suis, Pseudomonas aeruginosa, ß-hemolytic Escherichia coli and Staphylococcus aureus. The orange crystals and yellow precipitates were obtained from the preparation processes of the seed extracts. The orange-red crystals from the seeds collected from Lampang province exhibited strong in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging effects (EC50 value = 25.99 ± 3.30 µg/mL) and antibacterial effects on S. intermedius and ß-hemolytic E. coli while the yellow precipitate from the same source exhibited only antioxidant activity. Quantitative analysis of phytochemicals in O. indicum samples by spectrophotometric and HPLC techniques showed that they contained different amounts of total phenolic, total flavonoid and three major flavones; baicalin, baicalein and chrysin contents. Young fruit extract, which contained low amounts of flavone contents, still promoted antibacterial effects against the tested bacteria with IC50 values lower than 1 mg/mL and MIC values between 4 to 10 mg/mL in S. intermedius, S. aureus and S suis while higher IC50 and MIC values against P. aeruginosa and ß-hemolytic E. coli were found. From scanning electron microscopy, the extract of the young fruit of O. indicum promoted morphological changes in the bacterial cells by disrupting the bacterial cell walls, inducing leakage of the cellular content, and generating the abnormal accumulation of cells. The mechanism of action of the extract for this antibacterial effect may be the disruption of the cell membrane and abnormal cell aggregations. Regression analysis of the results suggests the correlation between total phenolic and total flavonoid contents and antioxidant and antibacterial effects. Baicalin was found to have a high correlation with an inhibitory effect against ß-hemolytic E. coli while three unidentified peaks, which could be flavones, showed high correlations with an inhibitory effect against S. intermedius, S. suis, P. aeruginosa and S. aureus.

In vitro antiplatelet activity of extract and its fractions of Paulownia Clone in Vitro 112 leaves.

BACKGROUND: Paulownia Clone in Vitro 112, also known as Oxytree is a hybrid of Paulownia elongata and Paulownia fortunei, developed under laboratory conditions. Its seeds are sterile, making it a noninvasive variety that can only be propagated in the laboratory. In China, species from the Paulownia genus (Paulowniaceae) are widely used in traditional medicine for the treatment of infectious diseases, such as gonorrhea and erysipelas. It has a broad spectrum of bioactivity, including neuroprotective, antioxidant, antibacterial, antiphlogistic, antiviral, and cytotoxic actions. However, the antiplatelet potential of Paulownia Clone in Vitro 112 has not yet been described. STUDY DESIGN: The aim of our study was thus to examine the effect of an extract and four fractions from leaves of Paulownia Clone in Vitro 112 on various parameters of platelet activation in an in vitro model. METHODS: Composition of the investigated extract and fractions was determined by UHPLC-UV-MS. The following parameters of platelet activation were investigated: nonenzymatic lipid peroxidation in resting platelets; enzymatic lipid peroxidation (AA metabolism) in platelets activated by thrombin; superoxide anion (O2-.) generation in the resting and activated platelets; platelet adhesion to collagen type I and fibrinogen; platelet aggregation stimulated by various physiological agonists, such as ADP, collagen, and thrombin. The effect of the extract and fractions on extracellular LDH activity, a marker of cell damage, was also determined. RESULTS: Verbascoside a phenylethnanoid glycoside, was the main secondary metabolite of the extract from leaves of oxytree (constituting approximately 45 % of all compounds). There were also iridoids, such as catalpol, aucubin, and 7-hydroxytomentoside, as well as flavonoids, such as luteolin and apigenin glycosides. Moreover, the extract had stronger antiplatelet properties than the fractions. For example, the extract at 10 µg/mL inhibited five parameters of platelet activation. CONCLUSIONS: Our results show that Paulownia Clone in Vitro 112 leaves are a new valuable source of compounds with antiplatelet potential.

Biotechnologically-Produced Myconoside and Calceolarioside E Induce Nrf2 Expression in Neutrophils.

The pathological manifestation of various diseases can be suppressed by the activation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2), a transcriptional regulator of the cellular redox balance. Haberlea rhodopensis Friv. is a resurrection plant species endemic for Bulgaria, containing biologically active phenylethanoid glycosides that might possess antioxidant or redox activity. This study aimed to analyze the metabolic profile of in vitro cultured H. rhodopensis and to identify molecules that increase Nrf2 expression in bone marrow neutrophils. Fractions B, D, and E containing myconoside, or myconoside and calceolarioside E in ratios 1:0.6 and 0.25:1 were found to be the most active ones. Fraction B (200 µg/mL) improved neutrophil survival and strongly increased the Nrf2 intracellular level, while D and E, as well as, myconoside and calceolarioside E at the same ratios had a superior effect. Calceolarioside E (32 µg/mL) had stronger activity than myconoside, the effect of which was very similar to that of 2-cyano-3,12-dioxo-oleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me), used as a positive control. These data indicate that both molecules, used alone or in combination have stimulatory activity on the endogenous Nrf2 level, indicating their therapeutic potential to regulate the cellular redox homeostasis oxidative stress-associated pathologies.

Pressurized Solvent Extraction of Paulownia Bark Phenolics.

Paulownia bark is mostly utilized jointly with wood, but the possibility of a separate valorization through the pressurized extraction of bark bioactives has been assessed. Subcritical water extraction and supercritical CO2 extraction are green technologies allowing shorter times than conventional solvent extraction under atmospheric shaken conditions. Subcritical water extraction was carried out at temperatures ranging from 140 to 240 °C and supercritical CO2 extraction was performed at different pressures (10, 20 and 30 MPa), temperatures (35, 45 and 55 °C) and ethanol concentrations (0, 10 and 15% (w/w)). Subcritical water extraction under a non-isothermal operation during heating up to 160 °C (19 min) provided extraction yields up to 30%, and the extracts contained up to 7% total phenolics with an ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging capacity equivalent to 35% the activity of Trolox, whereas at 240 °C, the yield decreased to 20%, but the phenolic content reached 21%, and the antiradical activity was equivalent to 85% of Trolox. Supercritical CO2 extraction at 30 MPa, 45 °C and 30 min reached a global yield of 2% after 180 min of extraction, but the product showed very low antiradical capacity. Gallic acid, vanillic acid, vanillin and apigenin were the major phenolic compounds found in the extracts.

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).

Experimental Data Based Machine Learning Classification Models with Predictive Ability to Select in Vitro Active Antiviral and Non-Toxic Essential Oils.

In the last decade essential oils have attracted scientists with a constant increase rate of more than 7% as witnessed by almost 5000 articles. Among the prominent studies essential oils are investigated as antibacterial agents alone or in combination with known drugs. Minor studies involved essential oil inspection as potential anticancer and antiviral natural remedies. In line with the authors previous reports the investigation of an in-house library of extracted essential oils as a potential blocker of HSV-1 infection is reported herein. A subset of essential oils was experimentally tested in an in vitro model of HSV-1 infection and the determined IC50s and CC50s values were used in conjunction with the results obtained by gas-chromatography/mass spectrometry chemical analysis to derive machine learning based classification models trained with the partial least square discriminant analysis algorithm. The internally validated models were thus applied on untested essential oils to assess their effective predictive ability in selecting both active and low toxic samples. Five essential oils were selected among a list of 52 and readily assayed for IC50 and CC50 determination. Interestingly, four out of the five selected samples, compared with the potencies of the training set, returned to be highly active and endowed with low toxicity. In particular, sample CJM1 from Calaminta nepeta was the most potent tested essential oil with the highest selectivity index (IC50 = 0.063 mg/mL, SI > 47.5). In conclusion, it was herein demonstrated how multidisciplinary applications involving machine learning could represent a valuable tool in predicting the bioactivity of complex mixtures and in the near future to enable the design of blended essential oil possibly endowed with higher potency and lower toxicity.