Photodynamic Inactivation of Human Coronaviruses.

Photodynamic inactivation (PDI) employs a photosensitizer, light, and oxygen to create a local burst of reactive oxygen species (ROS) that can inactivate microorganisms. The botanical extract PhytoQuinTM is a powerful photosensitizer with antimicrobial properties. We previously demonstrated that photoactivated PhytoQuin also has antiviral properties against herpes simplex viruses and adenoviruses in a dose-dependent manner across a broad range of sub-cytotoxic concentrations. Here, we report that human coronaviruses (HCoVs) are also susceptible to photodynamic inactivation. Photoactivated-PhytoQuin inhibited the replication of the alphacoronavirus HCoV-229E and the betacoronavirus HCoV-OC43 in cultured cells across a range of sub-cytotoxic doses. This antiviral effect was light-dependent, as we observed minimal antiviral effect of PhytoQuin in the absence of photoactivation. Using RNase protection assays, we observed that PDI disrupted HCoV particle integrity allowing for the digestion of viral RNA by exogenous ribonucleases. Using lentiviruses pseudotyped with the SARS-CoV-2 Spike (S) protein, we once again observed a strong, light-dependent antiviral effect of PhytoQuin, which prevented S-mediated entry into human cells. We also observed that PhytoQuin PDI altered S protein electrophoretic mobility. The PhytoQuin constituent emodin displayed equivalent light-dependent antiviral activity to PhytoQuin in matched-dose experiments, indicating that it plays a central role in PhytoQuin PDI against CoVs. Together, these findings demonstrate that HCoV lipid envelopes and proteins are damaged by PhytoQuin PDI and expands the list of susceptible viruses.

The Content of Phenolic Compounds and Organic Acids in Two Tagetes patula Cultivars Flowers and Its Dependence on Light Colour and Substrate.

The main focus of the study was to determine the content of phenolic acids, flavonoids, and organic acids in the flowers of Tagetes patula 'Petite Gold' and 'Petite Orange'. The growth of the plants was assessed depending on the cultivation conditions. The above plants were illuminated with white light, whereas the 'Petite Gold' ones with white light enhanced with blue or red light. Both cultivars grew in a two-level-mineral compounds organic substrate. The research showed that the French marigold flowers were rich in phenolic compounds and organic acids. The 'Petite Gold' flowers had more bioactive compounds compared with the 'Petite Orange' flowers. Three flavonoids, 10 phenolic acids and seven organic acids were found in the 'Petite Gold' flowers. The artificial lighting used during the cultivation of the plants showed diversified influence on the content of organic compounds in their flowers. The measurements of the plants' morphological traits and the number of inflorescences showed that illumination with red light resulted in a better effect. Large plants with numerous inflorescences grew in the substrate with a lower content of nutrients.

Red laser-mediated alterations in seed germination, growth, pigments and withanolide content of Ashwagandha [Withania somnifera (L.) Dunal].

Withania somnifera (L.) Dunal, generally well-known as Ashwagandha, is part of Indian traditional medicinal systems like Ayurveda, Siddha, and Unani for over 3000 years for treating an array of disorders. The chief bioactive component of this plant is the withanolides, a group of C28-steroidal lactone triterpenoids. These compounds are present in very low concentrations and hence cell culture methods have been used to enhance their production. Low-level laser irradiation has been reported to have elicited the seed germination, agronomical characters, biosynthesis of bioactive compounds in some plants. Therefore, the objective of the study was to investigate the effect of red (He-Ne) laser irradiation on seed germination, growth characters, pigment contents and withanolide content in W. somnifera. The seeds were inoculated onto two different combinations of Murashige and Skoog (MS) media and incubated for germination. The highest germination percentage was observed in ½ MS with pH 6.5 and GA3 presoaking followed by ½ MS with different pH. Four different doses of Helium-Neon (He-Ne) laser (10, 15, 20 and 25 J/cm2) were used to irradiate the seeds at 632.8 nm and germinated in vitro on ½ MS with pH 6.5. The maximum germination percentage, 63.88% was noted from seeds irradiated with 25 J/cm2 (P = 0.04). The highest total length of 13.33 cm was observed in the seedlings irradiated with 25 J/cm2 groups (P = 0.008). The highest total chlorophyll content of 329.5 µg/g fresh weight (FW) was observed for seedlings irradiated with 15 J/cm2 (P = 0.02) and the highest carotenoid content of 49.6 µg/g FW was observed for 25 J/cm2 treated seedlings. Further, primary root length was measured and found to be highest (11.14 cm) in seedlings irradiated with 10 J/cm2 and the highest number of lateral roots were observed for 15 and 25 J/cm2 groups. The significant amount of Withanolide A (WA) 0.52 µg/g dry weight (DW) and 0.60 µg/g DW was noted in 15 (P = 0.01) and 20 J/cm2 (P = 0.002) groups, respectively than control. The present investigation thus reveals the positive impact of red laser on the germination of seeds, growth characters and withanolide contents under in vitro environment.

Gamma irradiation for enhancing active chemical compounds in leaf extracts of Libidibia ferrea (Leguminosae).

This research was designed to evaluate the influence of the irradiation process of the leaf extracts of Libidibia ferrea (Leguminosae) on the production of secondary chemical compounds, including their biological activity. Leaves were collected and prepared to obtain the crude extract, which was then aliquoted and separately exposed to a Co-60 source with different doses, namely: 5, 7, 10, 12, 15, 20, 25, and 30 kGy. From irradiated and control samples, tests of toxicity were carried out with the microcrustacea Artemia salina Leach at three moments: 24 h, 60 and 180 days after the irradiation of the samples. Bioassays showed an increase in the toxicity of the irradiated extracts, correlated with the dose. The toxicity level did not change with the storage time, indicating the excellent stability of the samples. To assess the phytochemical profile of the crude and irradiated extracts, three techniques were employed: thin-layer chromatography (TLC), liquid chromatography coupled to mass spectrometry (LC-MS), and gas chromatography coupled to mass spectrometry (GC-MS). The phytochemical results emphasized the presence of phenols, tannins, and triterpenes. The analytical tests confirmed the role of ionizing radiation in breaking down macromolecules into simpler chemical species responsible for increasing chemical activity of the extract. This report presents and discusses ionizing radiation as an outstanding tool for enhancing active chemical compounds in leaf extracts of Libidibia ferrea, which reflects on their biochemical properties.

Photo-irradiation coupled biosynthesis of magnesium oxide nanoparticles for antibacterial application.

In recent times, magnesium oxide (MgO) nanoparticles are proven to be an excellent antibacterial agent which inhibits the growth of bacteria by generating reactive oxygen species (ROS). Release of ROS by nanoparticles will damage the cell membrane of bacteria and leads to the leakage of bacterial internal components and cell death. However, chemically synthesized MgO nanoparticles may possess toxic functional groups which may inhibit healthy human cells along with bacterial cells. Thus, the aim of the present study is to synthesize MgO nanoparticles using leaf extracts of Amaranthus tricolor and photo-irradiation of visible light as a catalyst, without addition of any chemicals. Optimization was performed using Box-Behnken design (BBD) to obtain the optimum condition required to synthesize smallest nanoparticles. The parameters such as time of reaction, the concentration of precursor, and light intensity have been identified to affect the size of biosynthesized nanoparticles and was optimized. The experiment performed with optimized conditions such as 0.001 M concentration of magnesium acetate as precursor, 5 cm distance of light (intensity), and 15 min of reaction time (light exposure) has led to the formation of 74.6 nm sized MgO nanoparticles. The antibacterial activities of MgO nanoparticles formed via photo-irradiation and conventional biosynthesis approach were investigated and compared. The lethal dosage of E. coli for photo-irradiated and conventional biosynthesis MgO nanoparticles was 0.6 ml and 0.4 ml, respectively. Likewise, the lethal dosage of S. aureus for both biosynthesis approaches was found to be 0.4 ml. The results revealed that the antibacterial activity of MgO nanoparticles from both biosynthesis approaches was similar. Thus, photo-irradiated MgO nanoparticles were beneficial over heat-mediated conventional method due to the reduced synthesis duration.

Survey of cytotoxic and UV protection effects of biosynthesized cerium oxide nanoparticles.

Cerium oxide nanoparticles (CeO2 NPs) are among the important nanoparticles that are extensively utilized in cosmetics, automotive industries, ultraviolet (UV) filtration, gas sensors, and pharmaceutical products. In this study, CeO2 NPs were synthesized using an aqueous extract of Ziziphus jujube fruit. The synthesized nanoparticles were characterized using UV-visible spectroscopy, powder X-ray diffraction, Fourier transform infrared spectroscopy, energy-dispersive spectroscopy, field energy scanning electron microscopy, and Raman methods. The results indicated that the size of synthesized nanoparticles is between 18 and 25 nm, and they have a spherical shape. UV absorbance of the synthesized nanoparticles was measured through spectrophotometric method in the range of 290 to 320 nm. The cytotoxic activity of synthesized CeO2 NPs against colon (HT-29) cancer cell line was surveyed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The results showed that synthesized nanoparticles are nontoxic on HT-29 cells under 400 µg/mL concentrations after 24 hours of treatment time periods. The increase in treatment time cases increases cytotoxic activity of synthesized nanoparticles. Sun protection factor of CeO2 NPs, as a criterion for amount of sunlight radiation protection, was determined by applying Mansur equation. The results demonstrated that synthesized CeO2 NPs have excellent UV protection and sunscreen physical absorption properties.

Water-Assisted/Water-Accelerated Photoreaction of trans-2,3,4',5-Tetrahydroxystilbene-2-O-ß-d-glucoside from the Roots of Polygonum multiflorum.

The photoreaction of 2,3,4',5-tetrahydroxystilbene-2-O-ß-d-glucoside (TSG) has been investigated. Water-assisted/water-accelerated photodimerization of trans-TSG favored the formation of syn-head-to-tail [2 + 2] photocyclobutane under 365 nm irradiation as a result of hydrophobic association and a fluorescent solute-solute aggregate from their excited singlet states. In contrast, irradiation with 254 nm led to [2 + 2] photocycloreversion. The two cyclobutane dimers were first obtained through straightforward photoreaction and identified as multiflorumiside A and multiflorumiside C through the detailed analysis of high-resolution electrospray ionization mass spectrometry and one- and two-dimensional nuclear magnetic resonance. Therefore, trans-TSG should be protected from light and water.

Soft Microwave Pretreatment to Extract P-Hydroxycinnamic Acids from Grass Stalks.

The aim of this article is to provide an analysis of microwave effects on ferulic and coumaric acids (FA and CA, respectively) extraction from grass biomass (corn stalks and miscanthus). Microwave pretreatment using various solvents was first compared to conventional heating on corn stalks. Then, microwave operational conditions were extended in terms of incident power and treatment duration. Optimal conditions were chosen to increase p-hydroxycinnamic acids release. Finally, these optimal conditions determined on corn stalks were tested on miscanthus stalks to underlie the substrate incidence on p-hydroxycinnamic acids release yields. The optimal conditions-a treatment duration of 405 s under 1000 W-allowed extracting 1.38% FA and 1.97% CA in corn stalks and 0.58% FA and 3.89% CA in miscanthus stalks. The different bioaccessibility of these two molecules can explain the higher or lower yields between corn and miscanthus stalks.

Sequential extraction of phenolics and pectin from mango peel assisted by ultrasound.

The present work evaluated the use of ultrasound to extract sequentially phenolics and pectin from mango peel. Initially, the influence of ethanol and ultrasound on the phenolics extraction was investigated. The results showed that the ultrasound did not affect the extraction yield of these compounds. The best total phenolics yield (67%) was obtained with an extraction solution consisting of 50% of ethanol in water (v/v) and without ultrasound application, according to the experimental design. As an innovative extraction methodology, the residue of this extraction was then used to extract pectin assisted by ultrasound. The use of ultrasound increased over than 50% of the pectin extraction yield and did not affect its quality, measured by the galacturonic acid content and the degree of esterification. The sequential extraction of phenolics and pectin shows to be an alternative to use the whole residue from mango peel.

Effects of ionizing radiation on bio-active plant extracts useful for preventing oxidative damages.

Humans are exposed to ionizing radiations in medical radiodiagnosis and radiotherapy that cause oxidative damages and degenerative diseases. Airplane pilots, and even more astronauts, are exposed to a variety of potentially harmful factors, including cosmic radiations. Among the phytochemicals, phenols are particularly efficient in countering the oxidative stress. In the present study, different extracts obtained from plant food, plant by-products and dietary supplements, have been compared for their antioxidant properties before and after irradiation of 140 cGy, a dose absorbed during a hypothetical stay of three years in the space. All the dry extracts, characterized in terms of vitamin C and phenolic content, remained chemically unaltered and maintained their antioxidant capability after irradiation. Our results suggest the potential use of these extracts as nutraceuticals to protect humans from oxidative damages, even when these extracts must be stored in an environment exposed to cosmic radiations as in a space station.

Dragon's Blood Sap: Storage Stability and Antioxidant Activity.

Currently, consumers are demanding additive-free, fresher, and more-natural products. Dragon's Blood Sap (DBS), the deep red latex of the specie of tree Croton lechleri (Müll. Arg.), contains a high concentration of phenolic compounds of great interest for the food, pharmaceutical, and cosmetic industries. These chemical compounds are highly susceptible to degradation. Therefore, DBS storage stability and its photo-oxidation was studied by Fourier transform infrared spectroscopy (FT-IR) and UV-Vis spectrophotometry for 39 days at different temperatures (4⁻21 °C) and relative humidities (0⁻56%), as well as under UV light exposure. It was observed that the degradation of phenolic compounds was reduced at 0% relative humidity (RH), not showing a significant effect of temperature in the range studied. UV light irradiation degraded DBS in a 20%. DBS has an exceptional high and stable antioxidant content (≥93% inhibition percentage of DPPH), which makes it a unique property to consider the DBS as an antioxidant agent or ingredient for consumer products formulations.

Antibacterial activity of irradiated extracts of Anacardium occidentale L. on multiresistant strains of Staphylococcus aureus.

This study evaluated the antibacterial activity of crude and fractionated leaf extracts of Anacardium occidentale, after receiving 10 kGy from 60Co, against multiresistant strains of Staphylococcus aureus in vitro. Minimum Inhibitory Concentrations-MIC and Minimum Bacteriostatic Concentrations-MBC were respectively assessed by serial microdilution technique in multiwall plates and Petri dishes, against standard strains and clinical isolates of multiresistant S. aureus. The results pointed out a significantly increase of the antibacterial activity of the such extracts after irradiation, emphasizing the role of gamma radiation on leaf extracts of A. occidentale to improve bioactive substances, offering new raw material for antibacterial drugs.

Ultrasound-assisted extraction of hemicellulose and phenolic compounds from bamboo bast fiber powder.

Ultrasound-assisted extraction of hemicellulose and phenolic compounds from bamboo bast fibre powder was investigated. The effect of ultrasonic probe depth and power input parameters on the type and amount of products extracted was assessed. The results of input energy and radical formation correlated with the calculated values for the anti-nodal point (λ/4; 16.85 mm, maximum amplitude) of the ultrasonic wave in aqueous medium. Ultrasonic treatment at optimum probe depth of 15 mm improve 2.6-fold the extraction efficiencies of hemicellulose and phenolic lignin compounds from bamboo bast fibre powder. LC-Ms-Tof (liquid chromatography-mass spectrometry-time of flight) analysis indicated that ultrasound led to the extraction of coniferyl alcohol, sinapyl alcohol, vanillic acid, cellobiose, in contrast to boiling water extraction only. At optimized conditions, ultrasound caused the formation of radicals confirmed by the presence of (+)-pinoresinol which resulted from the radical coupling of coniferyl alcohol. Ultrasounds revealed to be an efficient methodology for the extraction of hemicellulosic and phenolic compounds from woody bamboo without the addition of harmful solvents.

Enhanced UV protection of ketoconazole using Hyptis suaveolens micro emulsion.

Ketoconazole is photolabile antifungal drug. Photochemical reactions may decrease its therapeutic effect or induce toxic compounds. The aim of this study was to prepare ketoconazole loaded microemulsion containing H. suaveolens oil with antifungal and antioxidant powers in order to obtain effective antifungal formulation. The release study, antifungal activity and photostability test, were then evaluated. The results showed that optimized Hyptis suaveolens microemulsion for ketoconazole loading was selected through construction of pseudo-ternary phase diagrams. It consisted of 12.5% H. suaveolens oil, 12.5% capryol, 25% tween 80, 25% ethanol and 25% water. Mean globule size was 153 nm, as analyzed by photon correlation spectroscopy. Ketoconazole-loaded Hyptis suaveolens microemulsion and Hyptis suaveolens microemulsion had antifungal activity against Candida albican, Microsporum gypseum and Trichophyton mentagrophyte, showing inhibition zone ranged from 28-37 mm and 23-32 mm, respectively. Ketoconazole was released from Hyptis suaveolens microemulsion more than 90% within 5 days. In the results of photostability test, ketoconazole-loaded Hyptis suaveolens microemulsion gave significantly higher remaining ketoconazole than ketoconazole solution. This study demonstrated that Hyptis suaveolens microemulsion could be used to improve the photoprotection of photolabile drug.

Gamma-irradiated black ginseng extract inhibits mast cell degranulation and suppresses atopic dermatitis-like skin lesions in mice.

Gamma irradiation is able to affect various structural modification and an increase of the biological properties of biomaterials. This study was conducted to investigate the anti-allergenic effect of γ-irradiated black ginseng extract (BGE) using in vitro and in vivo experiments. IgEantigen complex-induced degranulation was measured in RBL-2H3 mast cells. In addition, an anti-atopic dermatitis (AD) test was carried out by spreading γ-irradiated BGE on the dorsal skin of 2,4-dinitrochlorobenzene (DNCB)-induced BALB/c mice. The content of arginylfructose (AF) of gamma-irradiated BGE was higher than that of BGE. In RBL-2H3 mast cells, γ-irradiated BGE treatments significantly reduced the IgE-antigen complex-induced release of ß-hexosaminidase, histamine, intracellular ROS, and Ca2+ influx. A western blot analysis showed that γ-irradiated BGE had an inhibitory activity on the FcεRI-mediated signaling in mast cells. In the DNCB-induced AD model, γ-irradiated BGE significantly alleviated the ADlike skin symptoms and clinical signs. The suppression of AD by γ-irradiated BGE was accompanied by a decrease in the serum level of IgE and IL-4, as well as the number of leukocyte. Gamma-irradiated BGE also suppressed IL-4 and increased IFN-γ in splenocytes. Our data suggests that γ-irradiated BGE may be effective therapeutic agents for the treatment of AD.

Decontamination of Mikania glomerata Leaves by Gamma Irradiation: Coumarin Determination by HPLC-DAD, Microbiological Control and Genotoxicological Studies.

Gamma irradiation as a decontaminating physical agent could be an important tool in the production chain of herbal medicines by improving the microbiological quality of raw materials and the safety of final products. This study was undertaken to investigate the genotoxic potential and eventual chemical modifications of a batch of Mikania glomerata raw material decontaminated by different doses of gamma irradiation (2.0, 3.5, and 5.0 kGy), using a cesium-137 source. DNA damage was assessed in vitro by agarose gel electrophoresis in regard to double-chain breaks of plasmid pUC 9.1 DNA and in vivo by micronucleus test in bone marrow cells of Wistar rats. Cytotoxicity in bone marrows was assessed by scoring polychromatic and normochromatic erythrocytes ratio. An HPLC-DAD method was adapted and validated for the enhancement of coumarin selectivity from the other matrix constituents. The microbial load was satisfactorily reduced, leading to sterilization at the highest dose. Genotoxic and cytotoxic effects were not increased in the in vitro and in vivo models. The concentration of coumarin and the chromatographic profiles of the hydroalcoholic plant extracts (ethanol 70% v/v) were not modified after such process. Therefore, this work suggests that gamma irradiation of M. glomerata raw material is suitable and safe for microbiological control purposes at the employed doses.

Is Gamma Radiation Suitable to Preserve Phenolic Compounds and to Decontaminate Mycotoxins in Aromatic Plants? A Case-Study with Aloysia citrodora Paláu.

This study aimed to determine the effect of gamma radiation on the preservation of phenolic compounds and on decontamination of dry herbs in terms of ochratoxin A (OTA) and aflatoxin B1 (AFB1), using Aloysia citrodora Paláu as a case study. For this purpose, artificially contaminated dry leaves were submitted to gamma radiation at different doses (1, 5, and 10 kGy; at dose rate of 1.7 kGy/h). Phenolic compounds were analysed by HPLC-DAD-ESI/MS and mycotoxin levels were determined by HPLC-fluorescence. Eleven phenolic compounds were identified in the samples and despite the apparent degradation of some compounds (namely verbasoside), 1 and 10 kGy doses point to a preservation of the majority of the compounds. The mean mycotoxin reduction varied between 5.3% and 9.6% for OTA and from 4.9% to 5.2% for AFB1. It was not observed a significant effect of the irradiation treatments on mycotoxin levels, and a slight degradation of the phenolic compounds in the irradiated samples was observed.

Characterization of the therapeutic properties of Chinese herbal materials by measuring delayed luminescence and dendritic cell-based immunomodulatory response.

Based on the traditional Chinese medicine theory, the Chinese pharmacopeia assigns a therapeutic description of "taste" to all herbs; thus, an herb's "taste" is valued in traditional Chinese medicine as a major ethnopharmacological category and reflects the herb's therapeutic properties. These properties guide the practitioner with respect to preparing a specific herbal formula in order to provide each patient with a personalized intervention. The key challenge in evidence-based medicine is to characterize herbal therapeutic properties from a multi-target, multi-dimensional systems pharmacology perspective. Here, we used delayed luminescence (DL, the slowly decaying emission of photons following excitation with light) as a rapid, direct, highly sensitive indicator to characterize the properties of herbal medicines. The DL parameters were able to reliably identify a specific category of herbal materials with the so-called "sweet" taste. To support the DL results and provide biological relevance to the DL results, we used a murine bone marrow-derived dendritic cell-based assay to examine the immunomodulatory effects of herbal extracts from various "taste" categories. Our results indicate that DL may serve as a robust and sensitive tool for evaluating the therapeutic properties of herbs based on the traditional Chinese medicine classification of "taste". Thus, DL provides a promising technological platform for investigating the properties of Chinese herbal medicines both qualitatively and quantitatively.

Effects of ultrasound pre-treatment on the structure of ß-conglycinin and glycinin and the antioxidant activity of their hydrolysates.

The effect of power, time and temperature of ultrasound on the structure of ß-conglycinin (7S) and glycinin (11S), and on the antioxidant activity of their hydrolysates were investigated. All ultrasound treated 7S and 11S fractions showed an increase in the α-helix and ß-turn proportions, and a decrease in ß-sheet and random coil proportions. The polarity of 7S and 11S microenvironment increased after ultrasound treatment. Ultrasound treatment significantly increased the reduction capacity and iron chelating capacity of 7S and 11S hydrolysates. The degree of hydrolysis and free SH groups of 7S and 11S hydrolysates increased after ultrasound pre-treatment. The relative content of high molecular weight peptides reduced, and the relative content of low molecular weight peptides increased in ultrasound treated 7S and 11S hydrolysates. The ultrasonication exposed certain groups of 7S and 11S fractions, improved contact with enzymes, and increased the content of highly active soybean antioxidant peptides.

Improving the antioxidant functionality of Citrus junos Tanaka (yuzu) fruit juice by underwater shockwave pretreatment.

Citrus junos Tanaka (yuzu) has a strong characteristic aroma, and hence, yuzu juice is used in a number of Japanese foods. We herein evaluated the functional compounds of yuzu juice to investigate whether underwater shockwave pretreatment affects its functionality. Employing the shockwave pretreatment at an increased discharge and energy of 3.5kV and 4.9kJ, respectively, resulted in an increase in the flavanone glycoside content and oxygen radical absorbance capacity (ORAC). The ORAC value of yuzu juice cultivated in Rikuzentakata increased approximately 1.7 times upon underwater shockwave pretreatment. The treatment method proposed herein exhibited reliable and good performance for the extraction of functional and antioxidant chemicals in yuzu fruits, and was comparable with traditional squeezing methods. The high applicability and reliability of this technique for improving the antioxidant functionality of yuzu fruit juice was demonstrated, confirming the potential for application to a wide range of food extraction processes.