Maximum Non-toxic Concentration Research Articles

Lantana undulata Schrank – A New Source of Biological Potential in the Verbenaceae Family

Background : The presence of condensed tannins, flavonones, flavonols, flavones, xantones, catechins and alkaloids in the ethanolic extract of Lantana undulata leaves has been associated with cytotoxicity and virucidal effect on suid herpesvirus. Objective: To evaluate cellular toxicity, antimicrobial and antioxidant activities, and anti-phlogistic property of the ethanolic extract leaves of Lantana undulata, a plant from the Verbenaceae family. Method: Cytotoxicity was evaluated on Madin Darby bovine kidney (MDBK) cells and erythrocytes and through leukocyte DNA damage. Antibacterial and antifungal activities were performed using the agar diffusion technique and broth microdilution. The inhibition percentage and viral inhibition index were established against bovine herpesvirus. The antioxidant effect (EC50) was determined by the DPPH technique and the mouse ear edema model was used to determine the anti-phlogistic activity. Results: The maximum nontoxic concentration for MDBK cells was 650 μg/ml. The ethanolic extract of L. undulata was considered non-hemolytic from 2 to 16 μg/ml and not genotoxic at the concentration tested (<8 μg/ml). High concentrations (3500 μg/ml) were needed to inhibit bacteria and yeasts, and the action of the extract was bacteriostatic or fungistatic. The IIV was of 0.37 and the IP was of 57.34% against bovine herpesvirus. The antioxidant EC50 was 540.87 μg/ml and the reduction of oedema occurred at 10, 30 and 100 mg/kg in the mouse model used. Conclusion: Although the ethanolic extract of L. undulataleaves showed to be potentially toxic and to have a reduced spectrum of action on microorganisms, the outstanding antiphlogistic action puts the therapeutic potential of this plant into perspective.

New experimental low-cost nanoscience technology for formulation of silver nanoparticles-activated carbon composite as a promising antiviral, biocide, and efficient catalyst

A simple low-cost one-pot photodeposition synthesis with no hazardous reactants or products is used to make silver nanoparticles-activated carbon composite (SNPs@AC). The SNPs are homogenously and photodeposited and absorbed into the activated carbon matrix. Both SNPs and SNPs@AC composite have particle sizes around 10 nm and 100 nm, respectively. The SNPs@AC composite showed good antiviral activity to VERO (ATCC ccl-81) cells. Zeta potential of SNPs@AC composite is −25 mV, showing that this colloidal system is electrically stable and resistant to coagulation. For many Gram-positive and Gram-negative bacteria, the SNPs@AC composite demonstrated strong antibacterial efficacy. The SNPs@AC composite has 75.72 percent anti-inflammatory effect at concentration 500 µg/mL. This composite has a maximum non-toxic concentration (MNTC) of 78.125 g/mL, which corresponds to antiviral activity of up to 96.7 percent against hepatitis A. virus (HAV). It is suggested as a candidate for pharmaceutical formulations, such as integration into the manufacture of N95 masks for COVID-19 infection protection. Concentration 160 μg/mL SNPs@AC composite has antioxidant activity 42.74% percent. The SNPs@AC composite exhibited selective catalytic activity for the organosynthesis hydrazination reaction of 4-chloro-3, 5-di-nitro-benzo-triflouride, giving 1-hydroxy-4-nitro-6-trifluoro-methyl benzotriazole, a common antiviral drug for severe acute respiratory syndrome (SARS). SNPs@composite's well-defined pores provide suitable active sites for binding reactants: 4-Cl-3, 5-di-NO2-benzotriflouride, and hydrazine, which react to create 1-hydroxy-4-nitro-6-trifluoromethyl benzotriazole, which diffuses into solution away from the catalyst surface, leaving the catalyst surface unaffected.

In Vitro Antiviral Activity of Potential Medicinal Plant Extracts Against Dengue and Chikungunya Viruses.

Dengue and chikungunya are two important mosquito-borne infections which are known to occur extensively in tropical and subtropical areas. Presently, there is no treatment for these viral diseases. In vitro antiviral screening of 25 extracts prepared from the plants of Vitex negundo, Plumeria alba, Ancistrocladus heyneanus, Bacopa monnieri, Anacardium occidentale, Cucurbita maxima, Simarouba glauca, and Embelia ribes using different solvents and four purified compounds (anacardic acid, chloroquinone, glaucarubinone, and methyl gallate) were carried out for their anti-dengue virus (DENV) and anti-chikungunya virus (CHIKV) activities. Maximum nontoxic concentrations of the chloroform, methanol, ethyl acetate, petroleum ether, dichloromethane, and hydroalcoholic extracts of eight plants were used. The antiviral activity was assessed by focus-forming unit assay, quantitative real-time RT-PCR, and immunofluorescence assays. Extracts from Plumeria alba, Ancistrocladus heyneanus, Bacopa monnieri, and Cucurbita maxima showed both anti-DENV and CHIKV activity while extract from Vitex negundo showed only anti-DENV activity. Among the purified compounds, anacardic acid, chloroquinone and methyl gallate showed anti-dengue activity while only methyl gallate had anti-chikungunya activity. The present study had identified the plant extracts with anti-dengue and anti-chikungunya activities, and these extracts can be further characterized for finding effective phytopharmaceutical drugs against dengue and chikungunya.

Picroside II alleviates DSS-induced ulcerative colitis by suppressing the production of NLRP3 inflammasomes through NF-κB signaling pathway

Context Ulcerative colitis (UC) is a common acute or chronic intestinal disease with an imbalance of inflammation. Picroside II (P-II) exerts a protective role in various inflammation-related diseases. However, the effect of P-II on UC is still unclear. Objective To explore the effect of P-II on UC and its potential mechanism. Materials and methods Human monocytic leukemia cell line THP-1 was treated with phorbol ester (PMA) to differentiate into a macrophage. The differentiated THP-1 cells were hatched with LPS combined with ATP or Nigericin to activate the NLRP3 inflammasome in vitro. The UC model was constructed by injection of DSS into mice. Results The maximum nontoxic concentration of P-II on THP-1 cells was 60 μM. LPS combined with ATP or Nigericin stimulated the production of IL-1β, which was antagonized by P-II treatment. Meanwhile, P-II administration interfered with the aggregation of ASC and the assembly of NLRP3 inflammasomes. Also, P-II treatment reduced the LPS and ATP-induced elevation of the relative protein expression of NLRP3, pro-caspase-1, IL-1β and p-p65/p65, and the concentration of TNF-α and IL-6. Besides, the NF-κB specific inhibitor BAY-117082 notably repressed the LPS together with ATP-enhanced the relative protein expression of NLRP3, caspase-1 and IL-1β. Moreover, in vivo results showed that P-II relieved the DDS-induced UC, as evidenced by the improvement of mice weight, DAI and pathological scores. In addition, P-II treatment notably decreased DDS-promoted expression of NLRP3 inflammasomes and inflammatory factors in vivo. Conclusion P-II alleviated DSS-induced UC by repressing the production of NLRP3 inflammasomes via the NF-κB signaling pathway.

Hypericum lanceolatum Lam. Medicinal Plant: Potential Toxicity and Therapeutic Effects Based on a Zebrafish Model.

Hypericum lanceolatum Lam. (H. lanceolatum) is a traditional medicinal plant from Reunion Island used for its pleiotropic effects mainly related to its antioxidant activity. The present work aimed to 1) determine the potential toxicity of the plant aqueous extract in vivo and 2) investigate its putative biological properties using several zebrafish models of oxidative stress, regeneration, estrogenicity, neurogenesis and metabolic disorders. First, we characterized the polyphenolic composition by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and identified chlorogenic acid isomers, quercetin and kaempferol derivatives as the major compounds. We then evaluated for the first time the toxicity of an aqueous extract of H. lanceolatum and determined a maximum non-toxic concentration (MNTC) in zebrafish eleutheroembryos from 0 to 96 hpf following OECD (Organization for Economic Cooperation and Development) guidelines. This MNTC test was also determined on hatched eleutheroembryos after 2 days of treatment (from 3 to 5 dpf). In our study, the anti-estrogenic effects of H. lanceolatum are supported by the data from the EASZY assay. In a tail amputation model, we showed that H. lanceolatum at its MNTC displays antioxidant properties, favors immune cell recruitment and tissue regeneration. Our results also highlighted its beneficial effects in metabolic disorders. Indeed, H. lanceolatum efficiently reduces lipid accumulation and body mass index in overfed larva- and adult-models, respectively. In addition, we show that H. lanceolatum did not improve fasting blood glucose levels in a hyperglycemic zebrafish model but surprisingly inhibited neurogenesis impairment observed in diabetic conditions. In conclusion, our study highlights the antioxidant, pro-regenerative, anti-lipid accumulation and pro-neurogenic effects of H. lanceolatum in vivo and supports the use of this traditional medicinal plant as a potential alternative in the prevention and/or treatment of metabolic disorders.

Screening of Antiviral Components of Yinhuapinggan Granule and Protective Effects of Yinhuapinggan Granule on MDCK Cells with Influenza A/H1N1 Virus.

Background Traditional Chinese medicine Yinhuapinggan granule (YHPG) has been used for treating upper respiratory tract infection like influenza, cough, and viral pneumonia. However, its active ingredients that really exert the main efficacy have not been well elucidated. This study is aimed at screening its antiviral components and investigating the potential therapeutic mechanisms of YHPG against the influenza A/PR8/34 (H1N1) virus in Madin Darby canine kidney (MDCK). Methods MDCK cells were infected with the influenza virus and then treated with ribavirin, YHPG, and main active ingredients in YHPG. Based on the maximum nontoxic concentration (TC0), half-maximal toxic concentration (TC50), half-maximal inhibitory concentration (IC50), and therapeutic index (TI), interferon-β (IFN-β) and interleukin-6 (IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA), and the gene expression of TLR7, MyD88, tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Jun amino terminal kinase (JNK), p38 mitogen-activated protein kinase (p38 MAPK), and p65 nuclear transcription factor-kappa B (p65 NF-κB) was quantified using reverse transcription-polymerase chain reaction (RT-PCR). Results The results indicated that the components of YHPG, such as ephedrine hydrochloride, pseudoephedrine hydrochloride, chlorogenic acid, and emodin, had significant antiviral effects. High and medium doses of YHPG effectively reduced the cytopathic effect (CPE) and significantly decreased IFN-β and IL-6 levels in the supernatant. Simultaneously, the transcript levels of TLR7, MyD88, TRAF6, JNK, p38 MAPK, and p65 NF-κB decreased in infected MDCK cells. Moreover, a certain dose-dependent relationship among different groups of YHPG was observed. Conclusions These results indicated that YHPG and the components of YHPG had a significant inhibitory function on the proliferation of the H1N1 virus. The mechanism might be associated with suppressing the activation of the TLR7/MyD88 signaling pathway, a decrease in the mRNA expression of key target genes, and inhibition of IFN-β and IL-6 secretion.

Zhilong Huoxue Tongyu Capsule Alleviated the Pyroptosis of Vascular Endothelial Cells Induced by ox-LDL through miR-30b-5p/NLRP3.

Background Our previous studies have demonstrated a protective role of Zhilong Huoxue Tongyu capsule in atherosclerosis (AS); however, the molecular mechanisms are unclear. Methods Human coronary artery endothelial cells (HCAECs) were induced with oxidized low-density lipoprotein (ox-LDL) to obtain cellular AS models. Then, the medicated serum of Zhilong Huoxue Tongyu capsule was obtained and used for treatment with ox-LDL-induced HCAECs. The cell viability was detected by CCK-8 assay. Besides, the binding between miR-30b-5p and NLRP3 was determined by the dual-luciferase reporter gene system assay. Furthermore, ox-LDL-induced HCAECs were transfected with miR-30b-5p mimic or miR-30b-5p inhibitor. The pyroptosis of HCAECs was assessed by flow cytometry, LDH content detection, and qRT-PCR assays. Results 10% medicated serum of Zhilong Huoxue Tongyu capsule was the maximum nontoxic concentration and it was used in subsequent assays. The rate of pyroptosis, LDH content, and the mRNA expression level of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were prominently enhanced after HCAECs were induced by ox-LDL, which were markedly rescued with medicated serum of Zhilong Huoxue Tongyu capsule. In addition, the medicated serum of Zhilong Huoxue Tongyu capsule significantly enhanced the ox-LDL-induced reduction of miR-30b-5p level. NLRP3 could bind to miR-30b-5p and was negatively corrected with miR-30b-5p. Moreover, all the rates of pyroptosis, LDH content, and the mRNA expression levels of pyroptosis-related genes including NLRP3, ASC, Caspase 1, IL-1β, and IL-18 were further observably decreased after ox-LDL-induced HCAECs treated with medicated serum were transfected with miR-30b-5p mimic, while these were significantly rescued with transfection of miR-30b-5p inhibitor. Conclusion Zhilong Huoxue Tongyu capsule alleviated the pyroptosis of vascular endothelial cells induced by ox-LDL through miR-30b-5p/NLRP3.

Green synthesized silver nanoparticles using Cyperus rotundus L. extract as a potential antiviral agent against infectious laryngotracheitis and infectious bronchitis viruses in chickens

BackgroundInfectious laryngotracheitis (ILT) and infectious bronchitis (IB) are two common respiratory diseases of poultry that inflict great economic burden on the poultry industry. Developing an effective agent against both viruses is a crucial step to decrease the economic losses. Therefore, for the first time green synthesized silver nanoparticles using Cyperus rotundus L. aqueous extract was evaluated in vitro as a potential antiviral against both viruses.ResultsSilver nanoparticles from Cyperus rotundus were characterized by the spherical shape, 11–19 nm size, and zeta potential of − 6.04 mV. The maximum nontoxic concentration (MNTC) was 50 µg mL−1 for both viruses without harmful toxicity impact. The study suggested that some of the compounds in C. rotundus extract (gallic acid, chlorogenic acid, and naringenin) or its silver nanoparticles could interact with the external envelope proteins of both viruses, and inhibiting extracellular viruses.ConclusionsThe results highlight that C. rotundus green synthesized silver nanoparticles could have antiviral activity against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) in chickens.Graphical

Antimicrobial and antiviral activities with molecular docking study of chitosan/carrageenan@clove oil beads.

Biopolymers are promising candidates that can be fabricated into hydrophilic matrices and used for many applications due to their distinctive properties such as non-toxic, biodegradable, biocompatibility, and low cost. A promising composite of chitosan and carrageenan with self-crosslinking has been prepared. In this study, a rigorous approach for an inexpensive and non-toxic combination of different amounts of clove oil with two polyelectrolytes including chitosan and carrageenan in the form of beads has been prepared. The structure and the surface morphology of the beads were investigated using FTIR, X-ray diffraction (XRD), and SEM. Moreover, antimicrobial, antiviral activity, and molecular docking were evaluated. Antibacterial results revealed that chitosan/carrageenan@clove oil beads have antimicrobial activity as well as chitosan/carrageenan without clove oil against Escherichia coli AATCC25922, Pseudomonas aeruginosa AATCC27853, Staphylococcus aureus ATCC25923, Bacillus subtilis AATCC6051, and Candida albicans ATCC90028. Furthermore, maximum non-toxic concentration (MNTC) of chitosan/carrageenan@clove oil beads was (31.25μg ml-1 ) which exhibited promising antiviral activity against Herpes simplex virus-1 (HSV-1), and was significantly higher than chitosan/carrageenan without clove oil, where antiviral activity was 82.94% and 57.64%, respectively. Eventually, docking study and computational calculation have been used to show the reactivity of the molecules. The developed chitosan/carrageenan@clove oil beads have shown promising properties to be used as carriers of drug delivery, tissue engineering, and regenerative medicine.

Curcumol inhibits encephalomyocarditis virus by promoting IFN-\u03b2 secretion

BackgroundEncephalomyocarditis virus (EMCV) infection can cause reproductive failure in sows and acute myocarditis and sudden death in piglets. It has caused huge economic losses to the global pig industry and that is why it is necessary to develop effective new treatment compounds. Zedoary turmeric oil has been used for treating myocarditis. Curcumol extracted from the roots of curcuma is one of the main active ingredient of zedoary turmeric oil. The anti-EMCV activity of curcumol along with the molecular mechanisms involved with a focus on IFN-β signaling pathway was investigated in this study.Method3-(4,5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the maximum non-toxic concentration (MNTC), 50% cytotoxic concentration (CC50), maximum inhibition rate (MIR) and 50% effective concentration (EC50) against EMCV. Through EMCV load, the anti-viral effect of curcumol was quantitatively determined using real-time quantitative PCR (qPCR). The effect of curcumol on the expression of IFN-β was investigated using real-time quantitative PCR and ELISA. Western blot was used to determine the amounts of MDA5, MAVS, TANK, IRF3 and P-IRF3 proteins in human embryonic kidney 293 T (HEK-293 T) cells infected with EMCV.ResultsThe results of MTT showed that compared with the ribavirin positive control group, the maximum inhibition ratio (MIR) of curcumol was greater but the selection index (SI) value was much smaller than that of ribavirin. The results of qPCR showed that curcumol and ribavirin significantly reduced the replication of EMCV in HEK-293 T cells. The curcumol (0.025 mg/mL) treatment has significantly increased IFN-β mRNA expression in the EMCV-infected HEK-293 T cells while ribavirin treatment did not. The results of ELISA showed that curcumol (0.025 mg/mL and 0.0125 mg/mL) has significantly increased the expression of IFN-β protein in EMCV-infected HEK-293 T cells. The results of Western blot showed that curcumol can inhibit the degradation of TANK protein mediated by EMCV and promote the expression of MDA5 and P-IRF3, while the protein expression level of MAVS and IRF3 remain unchanged.ConclusionCurcumol has biological activity against EMCV which we suggest that IFN-β signaling pathway is one of its mechanisms.

Effect of titanium dioxide nanoparticles on DNA methylation of human peripheral blood mononuclear cells.

The aim of the current study was to investigate the effect of well-characterized TiO2 nanoparticles on DNA methylation of peripheral blood mononuclear cells (PBMCs) in vitro. Maximum non-toxic concentration of nanoparticles for PBMCs was determined by MTT assay. The effect of TiO2 nanoparticles at concentrations of 25-100μg/ml on DNA methylation of PBMCs was investigated by measuring the %5-mC alterations through an ELISA assay. The physicochemical analysis showed that the TiO2 nanoparticles were crystalline, pure and in the anatase phase. Peaks related to Ti-O tensile vibrations were observed in the range of 1510cm-1. The size of nanoparticles was in the range of 39-74nm with an average hydrodynamic diameter of 43.82nm. According to the results of the MTT test, 100μg/ml was found to be maximum non-toxic concentration. The %5-mC in treated PBMCs revealed that TiO2 nanoparticles could lead to DNA hypomethylation in PBMCs. The %5-mC difference compared with the negative control was found to be 2.07 ± 1.02% (P=0.03). The difference of %5-mC between the 25 and 100μg/ml concentration of nanoparticles was statistically significant (P=0.02). The results of the current study show that the TiO2 nanoparticles cause DNA hypomethylation in PBMCs in a dose-response manner. Therefore, it is recommended to evaluate the effects of cytotoxicity and epigenotoxicity of commonly used nanoparticles before their use.

Abstract 3045: Investigating the influence of glycosylation using cellular models of neuroblastoma

Abstract Background: Neuroblastoma is a common childhood malignancy with approximately 800 new cases diagnosed each year in the United States. Treatment of neuroblastoma directly depends on patient risk categories and includes combinations of surgery, radiation therapy, chemotherapy, stem cell therapy, immunotherapy, and retinoid therapy. Various characteristics of cancer in general and some specific to neuroblastoma cell lines contribute to a difficulty in delivering clinically relevant concentrations of therapeutic agents to neuroblastoma masses. Amplification of the MYCN gene in certain neuroblastoma cell lines correlates with increased cellular surface glycosylation levels as well as a worse prognosis for patients. Overexpression of the MUC1 gene is also associated with increased glycosylation, specifically type-O, in various cancers, but the role of MUC1 in neuroblastoma has not been thoroughly defined. The objective of this study was to evaluate the influence of type-O glycosylation on therapeutic uptake in different neuroblastoma cell lines. Methods: The neuroblastoma cell line SK-N-DZ, a MYCN-amplified line, was compared to SK-N-SH, a non-MYCN-amplified line (negative control) and BT-20, a highly glycosylated breast cancer cell line (positive control). Benzyl-α-GalNAc, an O-glycosylation inhibitor, was introduced to each cell line. To determine glycosylation levels in the respective cell lines, fluorescence assays were performed using CD227 (anti-MUC1) antibody. In vitro cytotoxicity assays were performed to determine maximum non-toxic concentrations of the glycosylation inhibitor (benzyl-α-GalNAc) and antibody (CD227) and to subsequently evaluate the functional role of glycosylation using cellular models of neuroblastoma. Results: The maximum non-toxic concentrations per 10,000 cells for each neuroblastoma cell line was 0.05 mg/ml of benzyl-α-GalNAc, and 6 µl of CD227 stock preparation. Preliminary results from fluorescence detection studies showed an increase in CD227-FITC antibody uptake by the MYCN-amplified neuroblastoma cells (SK-N-DZ) following exposure to the O-glycosylation inhibitor (benzyl-α-GalNAc). Cellular uptake of CD227-FITC by non-MYCN-amplified cell line (SK-N-SH) was less by comparison. Conclusion: Preliminary findings suggest that O-glycosylation is a barrier capable of limiting cellular uptake of agents in MYCN-amplified neuroblastoma cell lines. Ongoing studies are focused on investigating the influence of glycosylation inhibition on cytotoxicity of FDA-approved therapeutic agents. Citation Format: Meghan Cook, Alma Garcia, Katie Konieczny, Meghan Ferguson, Robert Campbell. Investigating the influence of glycosylation using cellular models of neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3045.

Phytochemical investigation and differential effects of Cestrum elegans isolated compounds as antimicrobial and virucidal against hepatitis A virus

A new steroidal saponin, named as (25R)-6α-[(β-D-glucopyranosyl) oxy]-5α-spirostan-3β-yl β-D-glucopyranosyl (1''→3')-O-β-D-glucopyranosid (2) was isolated from the flowers of Cestrum elegans. In addition, a known steroidal saponin (1), a flavonoid compound (3) along with a triterpene saponin (4) were isolated for the first time in genus Cestrum and identified based on their chromatographic properties, chemical and spectral data (HR-ESI-MS, 1H, 13C NMR, 1H-1H COSY, HSQC, and HMBC). The four compounds are characterized by different effects against Gram positive, Gram negative bacteria, antifungal and antiviral activities. The maximum non-toxic concentration (MNTC) on vero cell line was 1.56 µg/mL for all compounds. While compound 1 and 3 showed the highest biological activities against hepatitis A virus possess 34.3% and 25% antiviral activity respectively..

Assessment of Antiviral Activity of Curcuma longa on Two RNA Viruses

Major pandemics and seasonal epidemics that have ravaged the world in the past and even at present, are mostly caused by RNA viruses. This has necessitated the need for continuous research to identify important natural products, with antiviral potentials, which can be harnessed for use in the prevention and treatment of viral infections. This study therefore, evaluated the antiviral property of Curcuma longa on two important RNA viruses of public health importance, namely polio and measles viruses. Extraction of active ingredients from turmeric rhizomes was done with the use of Analar grade methanol and concentrated using rotary evaporator. Polio and measles viruses were isolated from their respective vaccines using Reed-Muench method. Infective doses of the viruses and toxicity profile of extract were determined. Confluent Vero cells were inoculated with the viruses at different dilutions of the extract, incubated and observed for 7 days. Methanol extract of Curcuma longa inhibited polio virus at the maximum non-toxic concentration (MNTC) of 0.031μg μL-1 and inhibitory concentration (IC50) of 0.067 μg μL-1 with selectivity index of 2.16. Inhibition by the extract was observed prior to infection with the viruses. Phytochemical analysis of the extract showed presence of terpenes, saponins, alkaloids, flavonoids, tannins, cardiac glycosides and phenol as the bioactive phytochemicals. This study has shown that curcuma longa has potent inhibitory activity, hence can be harnessed in the development of an effective antiviral agent against polio and measles viruses.

THE ANTI HRSV ACTIVITY OF Ferula halophila Peşmen AQUEOUS AND METHANOL EXTRACT BY MTT ASSAY

The trend towards natural products in the world is increasing due to the increased drug resistance of infectious diseases, the high prices for drugs and the difficulty of access. Also, bacterial or viral diseases that are difficult to treat and need a long time for proper treatment cause important infections in people. Since effective drugs could not be developed for most viral infections, it is very important to find natural products against viruses to introduce them to the world of science. The antiviral activities of the aqueous and methanol extract from Ferula halophila Peşmen, an endemic species to Turkey, has been investigated against human respiratory syncytial virus (HRSV, ATCC-VR-26). The cytotoxic and antiviral properties of plant extracts were investigated in HRSV / HEp-2 cell systems, respectively by the colorimetric MTT assay. In the study, the titer of RSV was used as 100 TCID50 (50% tissue culture infective dose). While MNTC (Maximum non-toxic concentration) of methanol extract was 195.313 µg/mL and CC50 (50% cytotoxic concentration) was 4366.22 µg/mL, it was determined as MNTC = 390.625 µg/mL and CC50 = 4366.22 for aqueous extract. According to the results, methanol extract showed more toxicity than the aqueous extract. The MNTC of Ribavirin used as a positive control was determined as 0.98 µg/mL and CC50 was 110.40 µg/mL.As a result of the study, it was determined that the methanol extract was not effective, whereas the aqueous extract had a significant anti-HRSV activity with the values of 50.69 µg/mL EC50 (50% Effective Concentration) and 97.54 Selectivity Index (SI). The EC50 and SI values of Ribavirin were 2.39 µg/mL and 46.19, respectively.According to the results, we can explain the presence of antiviral activity only in the aqueous extract and the absence in the methanol extract by the high toxicity of it and the insolubility of the antiviral compounds in the methanol extract.

Drug Repurposing Patent Applications October-December 2020.

Drug Repurposing Patent Applications October-December 2020.

Antiviral activity of castor oil plant (Ricinus communis) leaf extracts

Ethnopharmacological relevanceRicinus communis L., commonly known as castor oil plant, is a precious traditional medicine with a history of thousands of years in the world. Castor oil plant has high traditional and medicinal values for treating liver infections, stomach ache, flatulence, constipation, inflammation, warts, colic, enteritis, fever, headache, and as a counter irritant. Its diverse phytochemicals have a wide range of valuable medicinal activities including hepatoprotective, anti-nociceptive, antioxidant, antiulcer, anticancer, anti-inflammatory, central analgesic, antidiabetic, antimicrobial, antiviral, and wound healing activity. Aim of the workTo provide a complete characterization of the composition of Ricinus communis leaves using ultra-performance liquid chromatography coupled with hybrid triple time-of-flight mass spectrometry (UPLC-Triple TOF-MS/MS) and different chromatographic techniques and to evaluate its antiviral potential using three mechanisms against three common viruses. Materials and methodsR. communis leaves were extracted with 70% methanol and further partitioned with solvents of increasing polarities: petroleum ether, dichloromethane (CH2Cl2), ethyl acetate, and n-butanol. The CH2Cl2 and n-butanol fractions were subjected to repeated chromatographic separation to isolate the phytochemicals, and their structures were elucidated using nuclear magnetic resonance spectroscopy. UPLC-Triple TOF-MS/MS was performed to determine the different phytochemicals in the ethyl acetate fraction. The antiviral activity of the extracts was investigated using the maximum nontoxic concentration of each against the challenge dose of the virus (CDV) and 1/10 and 1/100 dilutions of the CDV for Coxsackie B virus type 4 (COXB4), herpes simplex virus type 1 (HSV1), and hepatitis A virus (HAV) using Vero cell cultures that were treated according to three protocols to test for anti-replicative, protective, and anti-infective antiviral activity. Cell viability was evaluated using the MTT colorimetric assay and each experiment is repeated three times independently of each other. ResultsR. communis leaves possessed antiviral activity. Evaluation of the anti-replicative activity showed that all extracts possessed high anti-replicative activity against HAV especially methanol and methylene chloride fractions and moderate activity against COXB4; butanol > methylene chloride and ethyl acetate > methanol. All extracts showed protective activity against HAV, especially butanol extract, while methanol extracts showed higher non-significant antiviral protective activity against HSV1 vs Acyclovir. Almost no anti-infective effects were recorded for any extract against the studied viruses. ConclusionThe discriminatory effect against each virus by different mechanisms suggests the presence of different chemical compounds. The alkaloid and phenolic derivatives of the extracts of R. communis leaves may help develop a drug to prevent or treat common viral infections. Further investigations are recommended to define the bioactive antiviral properties of R. communis leaves.

Assessment of Antibacterial, Antifungal, Antioxidant and Antiviral Activities of Black pepper Aqueous Seed Extract

Traditional plant - based compounds are seeking more attention in pharmaceutical aspects to treat untreatable diseases because they have no side effect, easy recovery in great amount and a high degree of activity. Spices are extensively used to improve the taste and flavor of food, acquire a broad range of medicinal properties and some of them are known to be antioxidants. The antibacterial activity of aqueous seeds extract of Piper nigrum L. was evaluated against some pathogenic bacteria. The maximum inhibition zone was accounted against Gram negative bacteria Klebsiella oxytoca (21± 0.11mm) and minimum inhibition zone was exhibited against Gram positive bacteria Enterococcus faecalis (7 ± 0.05mm) while has no activity against Pseudomonas aeruginosa. Moreover, the aqueous seeds extract completely inhibits Penicillium citrinum growth at 500 μl by using food-poisoning technique and also showed some activity in controlling the mycelial growth of Aspergillus niger and Aspergillus flavus. Aqueous seeds extract of Piper nigrum L. was evaluated against Herpes simplex virus (HSV1) and Coxsackie virus (COXB4) and the results showed that Piper nigrum exert (57.26 and 13.41%) inhibition of plaque of HSV1 and COXB4 respectively. Moreover, the maximum nontoxic concentration (MNTC) for black pepper aqueous seed extract was 2500 μg/ml. The extract shows antioxidant activities with IC50 = 155 μg/ml and total phenolic content of (194.05 mg GA/g extract). These results suggest that aqueous seeds extract of Piper nigrum have significant anti-viral, antibacterial, antifungal and antioxidant activity signifying its usage as accepted antimicrobial representative.

Chemical Composition, Antiviral and Cytotoxic Activities of Essential Oil from Cyperus rotundus Growing in Egypt: Evidence from Chemometrics Analysis

The essential oil of Egyptian Cyperus rotundus rhizomes was obtained by hydrodistillation and analyzed by GC/MS. The oil is composed mainly of oxygenated sesquiterpenes. Thirty-seven compounds were identified including, humulene epoxide (38.43 %) and caryophyllene oxide (21.03 %) as major components. Correlations between different C. rotundus essential oils from Egypt and other regions were studied based on its chemical composition through principal components analysis and agglomerative hierarchical clustering. The oil was tested for its antiviral activity against hepatitis A, herpes simplex type 1, and coxsackie viruses at non-cytotoxic concentration. The oil was also evaluated for its cytotoxicity on hepatic, breast, and prostate carcinomas. It showed a very weak activity against tested viruses with percent protection 7.98, 14.21 and 8.79 %, respectively at a maximum non-toxic concentration of 125 μg/mL and it exhibited moderate cytotoxicity against hepatic and breast cancer cell lines with IC50 values of 204.1±1.25 and 170.8±0.567 μg/mL, respectively. The oil did not exhibit any cytotoxicity against the prostate cancer cell line.

Hepatoprotective homoisoflavonoids from the fruits of Cucumis bisexualis.

Cucumis bisexualis has been widely served as a beneficial wild fruit in China. Four new homoisoflavonoids (1-4), together with eight known homoisoflavonoid derivatives (5-12) were isolated from the 90% EtOH extract of C. bisexualis, using repeated open column chromatography and preparative thin-layer chromatography. They were elucidated based on the analysis of spectral data and references from the fruits of C. bisexualis for the first time. These compounds (1-12) were evaluated for their in vitro inhibitory effects against the increase of aspartate aminotransferase and alanine aminotransferase levels in H2 O2 -induced HepG2 cells in the range of their maximum nontoxic concentration. Among them, compounds 3, 4, 8, and 9 exhibited certain hepatoprotective activities. PRACTICAL APPLICATIONS: Cucumis bisexualis A.M. Lu ＆ G.C. Wang is an edible and medical fruit with many functional properties. A detailed study was made to identify the bioactive constituents of C. bisexualis and four new homoisoflavonoids (1-4) and eight known homoisoflavonoid derivatives (5-12) were isolated from the fruits of C. bisexualis for the first time. Compounds 3, 4, 8, and 9 exhibited certain hepatoprotective activities. This study suggests that natural compounds isolated from C. bisexualis could be used as promising hepatoprotective agents.