Epigenetic modifiers and minerals as tools to diversify secondary metabolite production in fungi

Secondary metabolite production of fungi can be modified by different approaches, including epigenetic modifiers, culture-dependent methods, and genomic-based methods. In this study, secondary metabolite production was explored in the presence of epigenetic modifiers and minerals using a microscale fermentation approach. Thirteen fungi originally isolated from mushrooms and soils were grown in 96-well microtiter plates (MTPs) using 70% of potato dextrose broth (PDB) with the addition of epigenetic modifiers and minerals in different combinations and concentrations. All cultures were fermented at 10 °C or 28 °C for 2, 3, or 5 weeks and extracted by solid phase extraction. The resulting extracts were subjected to high-performance liquid chromatography (HPLC) and the chromatograms were analyzed on a qualitative and quantitative basis. In addition, major secondary metabolites from four fungi were identified as penicillic acid, patulin, pseurotin A, and javanicin. Epigenetic modifiers and minerals induce significant changes in the profile of the secondary metabolites. Their usage combined with microscale fermentation provides a cost-efficient tool for exploring fungal secondary metabolism

Epigenetic modifiers and minerals as tools to diversify secondary metabolite production in fungi

Secondary metabolite production of fungi can be modified by different approaches, including epigenetic modifiers, culture-dependent methods, and genomic-based methods. In this study, secondary metabolite production was explored in the presence of epigenetic modifiers and minerals using a microscale fermentation approach. Thirteen fungi originally isolated from mushrooms and soils were grown in 96-well microtiter plates (MTPs) using 70% of potato dextrose broth (PDB) with the addition of epigenetic modifiers and minerals in different combinations and concentrations. All cultures were fermented at 10 °C or 28 °C for 2, 3, or 5 weeks and extracted by solid phase extraction. The resulting extracts were subjected to high-performance liquid chromatography (HPLC) and the chromatograms were analyzed on a qualitative and quantitative basis. In addition, major secondary metabolites from four fungi were identified as penicillic acid, patulin, pseurotin A, and javanicin. Epigenetic modifiers and minerals induce significant changes in the profile of the secondary metabolites. Their usage combined with microscale fermentation provides a cost-efficient tool for exploring fungal secondary metabolism

Stimulatory effects of Lycium shawii on human melanocyte proliferation, migration, and melanogenesis: In vitro and in silico studies

There is no first-line treatment for vitiligo, a skin disease characterized by a lack of melanin produced by the melanocytes, resulting in an urgent demand for new therapeutic drugs capable of stimulating melanocyte functions, including melanogenesis. In this study, traditional medicinal plant extracts were tested for cultured human melanocyte proliferation, migration, and melanogenesis using MTT, scratch wound-healing assays, transmission electron microscopy, immunofluorescence staining, and Western blot technology. Of the methanolic extracts, Lycium shawii L. (L. shawii) extract increased melanocyte proliferation at low concentrations and modulated melanocyte migration. At the lowest tested concentration (i.e., 7.8 μg/mL), the L. shawii methanolic extract promoted melanosome formation, maturation, and enhanced melanin production, which was associated with the upregulation of microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (TRP)-1 and TRP-2 melanogenesis-related proteins, and melanogenesis-related proteins. After the chemical analysis and L. shawii extract-derived metabolite identification, the in silico studies revealed the molecular interactions between Metabolite 5, identified as apigenin (4,5,6-trihydroxyflavone), and the copper active site of tyrosinase, predicting enhanced tyrosinase activity and subsequent melanin formation. In conclusion, L. shawii methanolic extract stimulates melanocyte functions, including melanin production, and its derivative Metabolite 5 enhances tyrosinase activity, suggesting further investigation of the L. shawii extract-derived Metabolite 5 as a potential natural drug for vitiligo treatment

Saudi female students learning English: Motivation, effort, and anxiety

Female students in Saudi Arabia who learn English as a foreign language (EFL) are increasing, and they may learn English for various purposes. These purposes may be broadly defined as integrative (focusing on understanding and affiliating with English-speakers) and instrumental (focusing on gaining pragmatic rewards, such as being accepted for university or getting a better job). Targeting 3rd year Intermediate (9th grade) and 3rd year Secondary (12th grade) female students, this study examined the relationship between these motivational constructs and effort, EFL anxiety, and differences between grades. Survey data from female students from two schools in Riyadh (N=200) were analysed using 2 (grade: 9th, 12th) x 2 (school) ANOVA. Results indicated that 12th graders were higher than 9th graders in instrumental motivation and effort, but lower in integrative motivation and anxiety. Differences between schools were small. Effort was positively correlated with integrative but not instrumental motivation for 9th graders, and was positively correlated with instrumental but negatively correlated with integrative motivation for 12th graders. Educators and curriculum designers should consider students' developmental needs to capitalize on their motivations in learning EFL

Saudi female students learning English : motivation, effort, and anxiety

Female students in Saudi Arabia who learn English as a foreign language (EFL) are increasing, and they may learn English for various purposes. These purposes may be broadly defined as integrative (focusing on understanding and affiliating with English-speakers) and instrumental (focusing on gaining pragmatic rewards, such as being accepted for university or getting a better job). Targeting 3rd year Intermediate (9th grade) and 3rd year Secondary (12th grade) female students, this study examined the relationship between these motivational constructs and effort, EFL anxiety, and differences between grades. Survey data from female students from two schools in Riyadh (N=200) were analysed using 2 (grade: 9th, 12th) x 2 (school) ANOVA. Results indicated that 12th graders were higher than 9th graders in instrumental motivation and effort, but lower in integrative motivation and anxiety. Differences between schools were small. Effort was positively correlated with integrative but not instrumental motivation for 9th graders, and was positively correlated with instrumental but negatively correlated with integrative motivation for 12th graders. Educators and curriculum designers should consider students' developmental needs to capitalize on their motivations in learning EFL

Natural Products as Novel Neuroprotective Agents; Computational Predictions of the Molecular Targets, ADME Properties, and Safety Profile

Neurodegenerative diseases (NDs) are one of the most challenging public health issues. Despite tremendous advances in our understanding of NDs, little progress has been made in establishing effective treatments. Natural products may have enormous potential in preventing and treating NDs by targeting microglia; yet, there have been several clinical concerns about their usage, primarily due to a lack of scientific evidence for their efficacy, molecular targets, physicochemical properties, and safety. To solve this problem, the secondary bioactive metabolites derived from neuroprotective medicinal plants were identified and selected for computational predictions for anti-inflammatory activity, possible molecular targets, physicochemical properties, and safety evaluation using PASS online, Molinspiration, SwissADME, and ProTox-II, respectively. Most of the phytochemicals were active as anti-inflammatory agents as predicted using the PASS online webserver. Moreover, the molecular target predictions for some phytochemicals were similar to the reported experimental targets. Moreover, the phytochemicals that did not violate important physicochemical properties, including blood-brain barrier penetration, GI absorption, molecular weight, and lipophilicity, were selected for further safety evaluation. After screening 54 neuroprotective phytochemicals, our findings suggest that Aromatic-turmerone, Apocynin, and Matrine are the most promising compounds that could be considered when designing novel neuroprotective agents to treat neurodegenerative diseases via modulating microglial polarization

<i>Limoniastrum monopetalum</i>–Mediated Nanoparticles and Biomedicines: In Silico Study and Molecular Prediction of Biomolecules

An in silico approach applying computer-simulated models helps enhance biomedicines by sightseeing the pharmacology of potential therapeutics. Currently, an in silico study combined with in vitro assays investigated the antimicrobial ability of Limoniastrum monopetalum and silver nanoparticles (AgNPs) fabricated by its aid. AgNPs mediated by L. monopetalum were characterized using FTIR, TEM, SEM, and DLS. L. monopetalum metabolites were detected by QTOF–LCMS and assessed using an in silico study for pharmacological properties. The antibacterial ability of an L. monopetalum extract and AgNPs was investigated. PASS Online predictions and the swissADME web server were used for antibacterial activity and potential molecular target metabolites, respectively. Spherical AgNPs with a 68.79 nm average size diameter were obtained. Twelve biomolecules (ferulic acid, trihydroxy-octadecenoic acid, catechin, pinoresinol, gallic acid, myricetin, 6-hydroxyluteolin, 6,7-dihydroxy-5-methoxy 7-O-β-d-glucopyranoside, methyl gallate, isorhamnetin, chlorogenic acid, 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-chromen-3-yl 6-O-(6-deoxy-β-l-mannopyranosyl)-β-d-glucopyranoside) were identified. The L. monopetalum extract and AgNPs displayed antibacterial effects. The computational study suggested that L. Monopetalum metabolites could hold promising antibacterial activity with minimal toxicity and an acceptable pharmaceutical profile. The in silico approach indicated that metabolites 8 and 12 have the highest antibacterial activity, and swissADME web server results suggested the CA II enzyme as a potential molecular target for both metabolites. Novel therapeutic agents could be discovered using in silico molecular target prediction combined with in vitro studies. Among L. Monopetalum metabolites, metabolite 12 could serve as a starting point for potential antibacterial treatment for several human bacterial infections

Adefovir Dipivoxil as a Therapeutic Candidate for Medullary Thyroid Carcinoma: Targeting RET and STAT3 Proto-Oncogenes

Aberrant gene expression is often linked to the progression of various cancers, making the targeting of oncogene transcriptional activation a potential strategy to control tumor growth and development. The RET proto-oncogene’s gain-of-function mutation is a major cause of medullary thyroid carcinoma (MTC), which is part of multiple endocrine neoplasia type 2 (MEN2) syndrome. In this study, we used a cell-based bioluminescence reporter system driven by the RET promoter to screen for small molecules that potentially suppress the RET gene transcription. We identified adefovir dipivoxil as a transcriptional inhibitor of the RET gene, which suppressed endogenous RET protein expression in MTC TT cells. Adefovir dipivoxil also interfered with STAT3 phosphorylation and showed high affinity to bind to STAT3. Additionally, it inhibited RET-dependent TT cell proliferation and increased apoptosis. These results demonstrate the potential of cell-based screening assays in identifying transcriptional inhibitors for other oncogenes

Exploring in vitro and in silico Biological Activities of Calligonum Comosum and Rumex Vesicarius: Implications on Anticancer and Antibacterial Therapeutics

Introduction: The adverse effects of clinically used anti-cancer medication and the rise in resistive micro-organisms have limited therapeutic options. Multiple anti-cancer drugs are derived from medicinal herbs which also have shown anti-bacterial effects. This study aimed to identify the optimal extraction solvent for detecting the cytotoxic and anti-bacterial effects of Calligonum comosum (C. Comosum) and Rumex vesicarius (R. Vesicarius) extracts. Additionally, the study aimed to identify active metabolites and assess their potential as future drug candidates for anti-cancer and anti-bacterial therapeutics. Methods: Leaves from both plants were extracted using ethanol, ethyl acetate, chloroform, and water. The cytotoxic effects of the extracts were tested on liver, colon, and breast cancer cell lines. Apoptosis was assessed using High Content Imaging (HCI) and the ApoTox triplex Glo assay. The anti-bacterial effects were determined using agar-well diffusion. Liquid chromatography-mass spectrometry (LC-MS) was used to tentatively identify the secondary metabolites. In silico computational studies were conducted to determine the metabolites' mode of action, safety, and pharmacokinetic properties. Results: The ethanolic extract of C. Comosum exhibited potent cytotoxicity on breast cancer cell lines, with IC50 values of 54.97 μg/mL and 58 μg/mL for KAIMRC2 and MDA-MB-231, respectively. It also induced apoptosis in colon and breast cancer cell lines. All tested extracts of C. Comosum and R. Vesicarius demonstrated anti-bacterial activity against Staphylococcus aureus and Escherichia coli. Seven active metabolites were identified, one of which is Kaempferol 3-O-Glucoside-7-O-Rhamnoside, which showed strong (predicted) anti-cancer activity. Kaempferol 3-O-Glucoside-7-O-Rhamnoside and Quercetin-3-O-Glucuronide also exhibited potential anti-bacterial effects on gram-positive and negative bacteria. Conclusion: Ethanol extraction of C. Comosum solubilizes active metabolites with potential therapeutic applications in cancer treatment and bacterial infections. Kaempferol 3-O-Glucoside-7-O-Rhamnoside, in particular, shows promise as a dual therapeutic drug candidate for further research and development to improve its efficacy, safety, and pharmacokinetic profile

Inactivation of Listeria monocytogenes in ready-to-eat smoked turkey meat by combination with packaging atmosphere, oregano essential oil and cold temperature

Abstract The effects of packaging atmosphere, storage temperature and oregano essential oil (EO) on growth of Listeria monocytogenes on ready-to-eat smoked turkey were studied. Smoked turkey slices were inoculated with a strain of Listeria monocytogenes Scott A (5.95, 5.28 and 5.26 log CFU/g) then vacuum packaged (VP), modified atmosphere packaging (MAP: 40% CO2 and 60% N2) and MAP with oregano essential oil (MAPEO), respectively. The treated slices were then stored at 0, 5, 10 and 15 °C for 179.88 days and the L. monocytogenes Scott A’s growth and microbial shelf life were monitored. The combination of MAP or MAPEO and storage temperature did not allow growth of L. monocytogenes higher than log 1 CFU/g during all storage periods. While in VP temperature combinations, the multiplication of bacteria were ≥ 1 log CFU/g. In VP, MAP and MAPEO smoked turkey, the growth of L. monocytogenes increased regardless of storage temperature. In MAPEO samples the inoculum in the product was suppressed by ca. 5 log CFU/g at 0, 10 and 15 °C at 180, 117 and 81 days of storage, respectively. The inhibition of L. monocytogenes in ready-to-eat smoked turkey by the combinations of MAP and MAPEO was enhanced by storage at 0 or 5 °C. The MAPEO system can be used effectively to control growth of pathogen in processed food when maintaining fixed temperature measures is difficult