Soybean-associated endophytic fungi as potential source for anti-COVID-19 metabolites supported by docking analysis.

AIMS: To identify the metabolites produced by the endophytic fungus, Aspergillus terreus and to explore the anti-viral activity of the identified metabolites against the pandemic disease COVID-19 in-silico. METHODS AND RESULTS: Herein, we reported the isolation of A. terreus, the endophytic fungus associated with soybean roots, which is then subcultured using OSMAC approach in five different culture media. Analytical analysis of media ethylacetate extracts using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was carried out. Furthermore, the obtained LC-MS data were statistically processed with MetaboAnalyst 4.0. Molecular docking studies were performed for the dereplicated metabolites against COVID-19 main protease (Mpro ). Metabolomic profiling revealed the presence of 18 compounds belonging to different chemical classes. Quinones, polyketides and isocoumarins were the most abundant classes. Multivariate analysis revealed that potato dextrose broth and modified potato dextrose broth are the optimal media for metabolites production. Molecular docking studies declared that the metabolites, Aspergillide B1 and 3a-Hydroxy-3, 5-dihydromonacolin L showed the highest binding energy scores towards COVID-19 main protease (Mpro ) (-9·473) and (-9·386), respectively, and they interact strongly with the catalytic dyad (His41 and Cys145) amino acid residues of Mpro . CONCLUSIONS: A combination of metabolomics and in-silico approaches have allowed a shorter route to search for anti-COVID-19 natural products in a shorter time. The dereplicated metabolites, aspergillide B1 and 3α-Hydroxy-3, 5-dihydromonacolin L were found to be potent anti-COVID-19 drug candidates in the molecular docking study. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that the endophytic fungus, A. terreus can be considered as a potential source of natural bioactive products. In addition to, the possibility of developing the metabolites, aspergillide B1 and 3α-Hydroxy-3, 5-dihydromonacolin L to be used as phytopharmaceuticals for the management of COVID-19.

Complementary Effect of Capparis Spinosa L. and Silymarin With/without Praziquantel on Mice Experimentally Infected with Schistosoma Mansoni.

Schistosomiasis remains to be the most common fibrotic disease resulting from inflammation and deposition of scar tissue around trapped parasitic eggs in the liver. Though chemotherapy eradicates matured worms efficiently and prevents the accumulation of schistosome eggs, fewer effective drugs are directed to reverse the present hepatic fibrosis. Therefore, treatment targeting hepatic fibrosis associated with schistosomiasis remains a challenging proposition. The present study was designed to investigate the potential complementary schistosomicidal and hepatoprotective activities of the methanol extract of Capparis spinosa L. (C. spinosa) with or without praziquantel (PZQ) and compare results with silymarin (Milk thistle), a known hepatoprotective and antifibrotic agent, on induced liver fibrosis by experimental Schistosoma mansoni (S. mansoni) infection. Total polyphenols in the extract were determined using colorimetric assay. C. spinosa L. caused a partial decrease in worm burden; a statistically significant reduction in hepatic and intestinal tissue egg load, what was associated histopathologically with decreasing in both the number and diameter of granulomas, as well as restoring serum aminotransferases (AST & ALT), alkaline phosphatase (ALP) and improving liver albumin synthesis. The best results were obtained in the group of mice treated with C. spinosa L. and PZQ together. Quantitative estimation of total polyphenols content using colorimetric assay showed that C. spinosa L. leaves contain higher concentration of polyphenolic compounds than fruits. It was concluded that C. spinosa L. has a promising hepatoprotective and antifibrotic properties and could be introduced as a safe and effective therapeutic tool with PZQ in the treatment of schistosomal liver fibrosis. Nevertheless further studies on the mechanism of action of C. spinosa L. in chronic liver diseases may shed light on developing therapeutic methods in clinical practice.

Solamargine production by a fungal endophyte of Solanum nigrum.

AIMS: The aim was to isolate, identify and characterize endophytes from Solanum nigrum L. as a new source of the cytotoxic steroidal alkaloid solamargine. METHODS AND RESULTS: Three endophytic fungi; SNFSt, SNFL and SNFF were isolated from S. nigrum and identified by molecular methods. Preliminary TLC screening showed a common metabolite between the plant and one of these fungi, SNFSt which was identified as Aspergillus flavus based on the phylogenetic analysis of its ITS sequence. Subsequent LC-HRESIMS analysis unambiguously established the identity of the compound based on its molecular formula and its characteristic MS(2) fragmentation pattern as solamargine. To ascertain its identity, fungal solamargine was isolated using preparative TLC and its structure was fully characterized using NMR spectroscopic techniques and high-resolution mass spectrometric analysis. Solamargine production could be followed and quantified for a total of 11 generations of this fungus with a titer of ~250-300 µg l(-1) . This study represents one of the first examples where host plant-derived compounds have been demonstrated to be steadily produced by an endophytic fungi in sizeable quantities. CONCLUSIONS: The production of solamargine (found in the host plant) by a cultivable fungal endophyte at a significant yield is a new observation. Further experiments such as media optimization, OSMAC (One Strain Many Compounds) or epigenetic modifiers could be applied to enhance the fungal solamargine production. SIGNIFICANCE AND IMPACT OF THE STUDY: The endophytic fungus SNFSt isolated from S. nigrum may be utilized for quantitative production of the potent cytotoxic metabolite solamargine.

Estrogenic Activity Including Bone Enhancement and Effect on Lipid Profile of Luteolin-7-O-glucoside Isolated from Trifolium alexandrinum L. in Ovariectomized Rats.

Luteolin-7-O-glycoside (LG), an abundant component in many edible plants, was found to be one of the major constituents of the aqueous methanol extract of Trifolium alexandrinum L. family Fabaceae, a fodder plant widely cultivated in Egypt. The estrogenic activity of LG concerning the effect on uterotrophy, lipid profile, weight gain and bone enhancement activity was determined in ovariectomized rat model at a dose of 5 mg/kg. Luteolin-7-O-glycoside showed significant estrogenic effect through the preservation of normal uterine weight and plasma estradiol level. It also significantly inhibited the bone turnover markers plasma bone-specific alkaline phosphatase, plasma osteocalsin, type I procollagen N-terminal, and C-telopeptide of type II collagen levels. It induced a significant improvement in plasma lipid profile. The effect of LG was comparable with estradiol with lower effect on uterine weight. Liver and kidney functions revealed a wide safety of LG at this dose level. The present study revealed that LG may be a promising hormone replacement therapy after being examined thoroughly on human. Copyright © 2016 John Wiley & Sons, Ltd.

Potential effect of methanol leaf extracts from three Podocarpus species on experimental cryptosporidiosis.

Cryptosporidiosis causes diarrhea in both immunocompetent and immunocompromised individuals, with acute manifestations occurring particularly in children and the elderly. Up till now, there is no curative therapy for cryptosporidiosis, so discovery of new classes of drugs are of great importance. This study aimed to examine the effect of methanol leaves extracts of the three Podocarpus species; P. macrophyllus (Thunb.), P. gracilior (Pilg.) and P. elongatus (Aiton) L' Hér. ex Pers and their combination on Cryptosporidium parvum (C. parvum) in experimentally infected mice in comparison with the commercially used drug, Nitazoxanide. As well as spectrophotometric estimation of the total phenolic and flavonoid content of these extracts was done. Results revealed that treatment with these three Podocarpus extracts and their combination showed a significant reduction of the number of C. parvum oocyst shed in the stool of infected mice compared to infected control group and Nitazoxanide- infected treated group at P < 0.001. The combination of the three Podocarpus extracts was the most effective treatment showing the lowest number of oocysts shedding in comparison with other used extracts and Nitazoxanide. Histopathological inspection of sections from ilium and colon displayed signs of improvement after treatment with P. macrophyllus and P. gracilior extracts and more remarkable improvement when the three extracts were combined. It was concluded that the three Podocarpus species extracts used in this study had a promising anti-Cryptosporidium activity especially when they were combined.

Phyto- and Bio-Chemical evaluation of Diospyros kaki L. cultivated in Egypt and its biological activities.

Diospyros kaki L. or Costata cultivar is the main persimmon variety progressively consumed in the Egyptian market and exportation. The objective of this study was to investigate the bioprotective effects of the alcoholic extract of fruits as well as leaves of D. kaki using in-vivo rat models. Petroleum ether extracts of fruits and leaves were analytically characterized for saponifiable and unsaponifiable compounds by GC/MS. Main flavonoids were chromatographically isolated from 80% aqueous methanol extract of leaves. Chemical evaluation for fruits and leaves namely, content of moisture, ash, proteins, fatty acids, amino acids, fat and water soluble vitamins, minerals, carotenoids was carried out. Total antioxidant activity was determined by radical scavenging effects using DPPH assay. The effect on biochemical parameters and its biological activity were also performed. Results revealed the identification of the major compounds of saponifiable and unsaponifiable matters of fruits and leaves. Scopoletin, as well as, kaempferol, luteolin, rutin and apigenin 7-O-glucoside were isolated and identified. It showed that this plant can provide a good nutritional value and it is safe regarding the kidney and liver functions, good source that help in enhancing the antioxidant defense against free radicals. No abnormal effects were found in lipids profile on experimental animals and there were good results in the ratio of HDL and LDL cholesterol. Also, this plant can help in optimizing blood sugar, enhancing the level of blood haemoglobin. It is concluded that D. kaki displays a good source of nutrients and bioactive compounds that may contribute to its therapeutic benefits against the risk of disease complications.

Eliciting callus culture for production of hepatoprotective flavonoids and phenolics from Sequoia sempervirens (D. Don Endl).

The aim of our study is to estimate the hepatoprotective effects of the ethanolic extract of the leaves of Sequoia sempervirens by determination of liver biomarkers (ALT, AST, total bilirubin and albumin in serum) and by histopathological examinations using thioacetamide-induced (TAA) liver injury model. Concurrent administration of ethanolic extracts of S. sempervirens leaves improved the alterations in liver morphology where it was a potent protector of the liver. The potential of L-phenylalanine and silver nitrate as chemical elicitors as well as UV radiation as a physical elicitor on flavonoid production in callus culture of S. sempervirens were emphasized. Murashige and Skoog's medium fortified with phenylalanine and silver nitrate enhanced the production of flavonoids and phenolics. HPLC analysis was performed for qualitative and quantitative estimation of some flavonoid compounds in the produced calli in comparison with the mother plant. This finding highlights the potential use of S. sempervirens in the treatment of liver dysfunction.

Phyto- and Bio-Chemical evaluation of Diospyros kaki L. cultivated in Egypt and its biological activities / Avaliação fito e bioquímica de Diospyros kaki L. cultivado no Egito e suas atividades biológicas

Abstract Diospyros kaki L. or Costata cultivar is the main persimmon variety progressively consumed in the Egyptian market and exportation. The objective of this study was to investigate the bioprotective effects of the alcoholic extract of fruits as well as leaves of D. kaki using in-vivo rat models. Petroleum ether extracts of fruits and leaves were analytically characterized for saponifiable and unsaponifiable compounds by GC/MS. Main flavonoids were chromatographically isolated from 80% aqueous methanol extract of leaves. Chemical evaluation for fruits and leaves namely, content of moisture, ash, proteins, fatty acids, amino acids, fat and water soluble vitamins, minerals, carotenoids was carried out. Total antioxidant activity was determined by radical scavenging effects using DPPH assay. The effect on biochemical parameters and its biological activity were also performed. Results revealed the identification of the major compounds of saponifiable and unsaponifiable matters of fruits and leaves. Scopoletin, as well as, kaempferol, luteolin, rutin and apigenin 7-O-glucoside were isolated and identified. It showed that this plant can provide a good nutritional value and it is safe regarding the kidney and liver functions, good source that help in enhancing the antioxidant defense against free radicals. No abnormal effects were found in lipids profile on experimental animals and there were good results in the ratio of HDL and LDL cholesterol. Also, this plant can help in optimizing blood sugar, enhancing the level of blood haemoglobin. It is concluded that D. kaki displays a good source of nutrients and bioactive compounds that may contribute to its therapeutic benefits against the risk of disease complications.

Resumo Diospyros kaki L. ou Costata é a principal variedade de caqui progressivamente consumida no mercado e exportação egípcia. O objetivo deste estudo foi investigar os efeitos bioprotetores do extrato alcoólico de frutos e folhas de D. kaki utilizando modelos de ratos "in vivo". Extratos de éter de petróleo provenientes de frutos e folhas foram caracterizados analiticamente para compostos saponificáveis ​​e insaponificáveis ​​por GC/MS. Os principais flavonoides foram isolados cromatograficamente, a partir de 80% de extrato aquoso de folhas de metanol. Foi realizada avaliação química de frutos e folhas, ou seja, de teor de umidade, cinzas, proteínas, ácidos graxos, aminoácidos, vitaminas lipossolúveis e solúveis em água, minerais, carotenoides. Foi caracterizada a atividade antioxidante, utilizando ensaio (DPPH). O efeito nos parâmetros bioquímicos e sua atividade biológica também foram analisados. Os resultados revelaram a identificação dos principais compostos saponificáveis e insaponificáveis de frutos e folhas. Escopoletina, bem como kaempferol, luteolina, rutina e apigenina 7-O-glicosídeo foram isolados e identificados. Também demonstraram que esta planta pode proporcionar um bom valor nutricional e é segura em relação às funções renais e hepáticas, boa fonte que ajuda a melhorar a defesa antioxidante contra os radicais livres. Não foram encontrados efeitos anormais no perfil lipídico em testes em animais, e houve um efeito satisfatório na relação de colesterol HDL e LDL. Além disso, esta planta pode ajudar na otimização do açúcar no sangue, aumentando o nível de hemoglobina. Conclui-se que o D. kaki apresenta uma boa fonte de nutrientes e de compostos bioativos que contribuem para seus benefícios terapêuticos e contra o risco de complicações de doenças.

Flavonoid constituents, cytotoxic and antioxidant activities of Gleditsia triacanthos L. leaves.

Gleditsia triacanthos L. is a deciduous tree belonging to the family Fabaceae. It possesses important biological activities as anti-mutagenic, anticancer, cytotoxic and treating rheumatoid arthritis. The total ethanol extract (EtOHE) and successive extracts (petroleum ether, chloroform, ethyl acetate, and aqueous ethanol) were prepared from the leaves. Eight flavone glycosides and two flavone aglycones named vicenin-I (1), vitexin (2), isovitexin (3), orientin (4), isoorientin (5), luteolin-7-O-ß-glucopyranoside (6), luteolin-7-O-ß-galactopyranoside (7), apigenin-7-O-ß-glucopyranoside (8), luteolin (9) and apigenin (10) were isolated from the aqueous ethanol extract of G. triacanthos L. leaves. Potent cytotoxic activity of the EtOHE extract was observed against the liver (IC50 = 1.68 µg), breast (IC50 = 0.74 µg), cervix (IC50 = 1.28 µg), larynx (IC50 = 0.67 µg) and colon (IC50 = 2.50 µg) cancer cell lines. Cytotoxic activity of compounds 2, 4, 6 and 8 against, the liver, breast and colon cancer cell lines was also proved. Evaluation of the in-vivo antioxidant activity of the EtOHE and successive extracts revealed that the highest activity was exhibited by 100 mg of EtOHE (97.89% potency) as compared with vitamin E (100% potency). Compound 6 showed 91.8% free radical scavenging activity.

Phyto- and Bio-Chemical evaluation of Diospyros kaki L. cultivated in Egypt and its biological activities

Abstract Diospyros kaki L. or Costata cultivar is the main persimmon variety progressively consumed in the Egyptian market and exportation. The objective of this study was to investigate the bioprotective effects of the alcoholic extract of fruits as well as leaves of D. kaki using in-vivo rat models. Petroleum ether extracts of fruits and leaves were analytically characterized for saponifiable and unsaponifiable compounds by GC/MS. Main flavonoids were chromatographically isolated from 80% aqueous methanol extract of leaves. Chemical evaluation for fruits and leaves namely, content of moisture, ash, proteins, fatty acids, amino acids, fat and water soluble vitamins, minerals, carotenoids was carried out. Total antioxidant activity was determined by radical scavenging effects using DPPH assay. The effect on biochemical parameters and its biological activity were also performed. Results revealed the identification of the major compounds of saponifiable and unsaponifiable matters of fruits and leaves. Scopoletin, as well as, kaempferol, luteolin, rutin and apigenin 7-O-glucoside were isolated and identified. It showed that this plant can provide a good nutritional value and it is safe regarding the kidney and liver functions, good source that help in enhancing the antioxidant defense against free radicals. No abnormal effects were found in lipids profile on experimental animals and there were good results in the ratio of HDL and LDL cholesterol. Also, this plant can help in optimizing blood sugar, enhancing the level of blood haemoglobin. It is concluded that D. kaki displays a good source of nutrients and bioactive compounds that may contribute to its therapeutic benefits against the risk of disease complications.

Resumo Diospyros kaki L. ou Costata é a principal variedade de caqui progressivamente consumida no mercado e exportação egípcia. O objetivo deste estudo foi investigar os efeitos bioprotetores do extrato alcoólico de frutos e folhas de D. kaki utilizando modelos de ratos in vivo. Extratos de éter de petróleo provenientes de frutos e folhas foram caracterizados analiticamente para compostos saponificáveis e insaponificáveis por GC/MS. Os principais flavonoides foram isolados cromatograficamente, a partir de 80% de extrato aquoso de folhas de metanol. Foi realizada avaliação química de frutos e folhas, ou seja, de teor de umidade, cinzas, proteínas, ácidos graxos, aminoácidos, vitaminas lipossolúveis e solúveis em água, minerais, carotenoides. Foi caracterizada a atividade antioxidante, utilizando ensaio (DPPH). O efeito nos parâmetros bioquímicos e sua atividade biológica também foram analisados. Os resultados revelaram a identificação dos principais compostos saponificáveis e insaponificáveis de frutos e folhas. Escopoletina, bem como kaempferol, luteolina, rutina e apigenina 7-O-glicosídeo foram isolados e identificados. Também demonstraram que esta planta pode proporcionar um bom valor nutricional e é segura em relação às funções renais e hepáticas, boa fonte que ajuda a melhorar a defesa antioxidante contra os radicais livres. Não foram encontrados efeitos anormais no perfil lipídico em testes em animais, e houve um efeito satisfatório na relação de colesterol HDL e LDL. Além disso, esta planta pode ajudar na otimização do açúcar no sangue, aumentando o nível de hemoglobina. Conclui-se que o D. kaki apresenta uma boa fonte de nutrientes e de compostos bioativos que contribuem para seus benefícios terapêuticos e contra o risco de complicações de doenças.