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Objective: Most research on the therapeutic effects of wild pistachio species has focused on bacteria and fungi. This study investigated the flavonoid and phenolic contents of different leaves and hulls of Pistacia atlantica (P. atlantica) subsp. Kurdica (P. atlantica subsp. Kurdica) extracts and their effect on herpes simplex virus 1 (HSV1). Methods: In this study, aqueous, ethanolic, and methanolic extracts of the leaf and hull of P. atlantica subsp. Kurdica were prepared by the percolation method. The total phenolic and flavonoid contents in different extracts were measured by UV/Vis spectrophotometry using the Folin-Ciocâlteu reagent and the colorimetric method of aluminum chloride, respectively. On African green monkey kidney cells (VERO), the ethanolic extract of P. atlantica subsp. Kurdica was tested for toxicity. Furthermore, 50% cytotoxic and inhibitory concentrations (CC50 and IC50) were identified. Results: Compared with aqueous and methanolic extracts, the ethanolic extract of the leaf and hull of P. atlantica subsp. Kurdica had the highest phenolic and flavonoid concentrations. In addition, our study showed that CC50 values were 661.81 and 795.21 μg/mL. Also, IC50 values were 97.51 and 110.82 μg/mL for leaf and hull extracts, respectively. The selectivity index for the ethanolic leaf and hull extracts were 6.79 and 7.18, respectively. Conclusion: Our results showed that P. atlantica subsp. Kurdica had an anti-HSV1 effect in a dose-dependent manner but at a higher dose than acyclovir. Ethanolic extract of P. atlantica subsp. Kurdica is probably a suitable herbal medicine with anti-herpetic effects. [ABSTRACT FROM AUTHOR]

Background. The COVID-19 pandemic, caused by the SARS-CoV-2 coronavirus, originated in Wuhan, China, has claimed millions of lives around the world. In this regard, the search for effective drugs, including the repurposing of existing ones, has become an urgent task. A promising treatment strategy appears to be drug disruption of viral reproduction. RNA-dependent RNA polymerase (RdRp) is the central subunit of the RNA synthesis process for all positive-strand RNA viruses and is therefore an attractive target for antiviral inhibitors.The aim of this work is an experimental study of the antiviral activity of various drugs based on 6-fluoro-3-hydroxy-2-pyrazinecarboxamide (Favipiravir) in vitro and in vivo against the SARS-CoV-2 coronavirus (COVID-19).Material and methods. The experiments were carried out on a permanent culture of African green monkey kidney cells — Vero Cl008. The effectiveness of the drugs was assessed by suppressing the reproduction of the virus in vitro. Biological activity was assessed by titration of the virus-containing suspension in Vero Cl008 cell culture by the formation of negative colonies. Syrian golden hamsters orally infected with the SARS-CoV-2 virus, variant B, were used in the study. The effectiveness of the drug was assessed by the coefficient of therapeutic action.Results. The results of the study revealed that the compounds FP-1 and Avifavir in the concentration range of 100–400 µg/ml almost completely suppress the reproduction of the SARS-CoV-2 virus; the CTI index for the drug FP-1 was 4, for Avifavir it was 2. The ED₅₀ value for FP-1 was 26 µg/ml, for Avifavir it was 36 µg/ml. Preparations T-705 and Coronavir revealed antiviral activity only at extremely high concentrations. The CTI was 1. During the study on Syrian golden hamsters orally infected with the SARS-CoV-2 virus, variant B, at a dose of 5×105 PFU, it was shown that the use of Avifavir and FP-1 has a high protective efficacy, while Coronavir and T-705 cause a moderate suppression of virus reproduction in the target organ. According to the complex of clinical-virological, biochemical, and hematological indicators, the disease severity index (DSI) and the therapeutic index (TI) were calculated. For the drug Avifavir, the DSI was 0.269; the TI was 71.3% with a probability of 99.9%.Conclusion. Of the studied compounds, Avifavir and FP-1 showed the highest antiviral activity.

Lung cancer ranks as the 2nd most common cancer globally. It’s the most prevalent cancer in men and the 2nd most common in women. The prominent events in EGFR-mutated non-small-cell lung cancer (NSCLC) include the emergence of the L858R mutation within EGFR exon 21. Despite the promising efficacy of EGFR inhibitors in managing lung cancer, the development of acquired resistance poses a significant hurdle. In the current investigation, we focused on the screening of two phytochemicals, namely Dehydrocostus lactone and Mokkolactone, derived from the Saussurea lappa plant, as potential inhibitors targeting EGFR L858R mutant lung cancer. The chloroform and ethanol extract of the plant demonstrated anti-proliferative activity through the Resazurin chemosensitivity assay, exhibiting an IC50 value of 37.90 ± 0.29 µg/ml with selectivity index 2.4. Through a GC–MS study, we identified 11 phytochemicals for further insilico analysis. These compounds underwent ADMET assessment followed by drug likeliness analysis before being subjected to molecular docking against EGFR L858R, identified through protein–protein interaction network analysis. All phytochemicals exhibited binding energy scores ranging from −6.9 to −8.1 kcal/mol. Dehydrocostus lactone and Mokkolactone were specifically identified for their binding profile. Findings from 100 ns molecular dynamics simulations demonstrated their enhanced stability compared to the reference ligand DJK. This was evident in the root mean square deviation (RMSD) values, ranging from 0.23 ± 0.01 nm to 0.30 ± 0.05 nm, the radius of gyration values, from 1.71 ± 0.01 nm to 1.72 ± 0.01 nm, and the solvent accessible surface area values, from 155.39 ± 2.40 nm2 to 159.32 ± 2.14 nm2. Additionally, favourable characteristics were observed in terms of hydrogen bonding, principal component analysis, and free energy landscape analysis. Examination of their electronic structure via density functional theory revealed efficient properties, with the highest occupied molecular orbital-least unoccupied molecular orbital energy gap values ranging from −3.984 eV to −6.547 eV. Further, in vivo analysis is required to gain a more comprehensive understanding and efficacy of these identified phytochemicals against lung cancer.

The aim of the work is to study the effect of new azoloazine derivatives on the level of DNA damage to Vero cells (DNA comet test) in vitro by alkaline gel electrophoresis.Material and methods. The objects of the study are 8-(piperidinocarbonyl)3-cyclohexylimidazo[5,1-d][1,2,3,5]tetrazine-4(3H)-one (1), diethyl ether 4-aminoimidazo[5,1-c][1,2,4]triazine3,8-dicarboxylic acid (2), 4-amino-8-ethoxycarbonylimidazo[5,1-c][1,2,4]triazine-3-N-(p-toluyl)carboxamide (3). Epirubicin was chosen as a comparison drug. The compounds were used in doses of 1/2, 1/10 and 1/50 IC50. The Vero cell line cultured according to the standard protocol was selected as the cell model. To assess genotoxicity, an alkaline version of the DNA comet method was used, which has a high sensitivity and allows detecting DNA damage.Results. Analysis of the data obtained indicates that the tested compounds 1-3 enhance DNA damage in non-tumor cells. Compound 1 has the most pronounced genotoxic effect. Thus, the use of this substance in a dose of ½ IC50 led to a significant increase in the length of the comet’s tail by 1.5 times. It was noted that DNA damage under the action of compound 1 in the studied doses and of epirubicin was on the same level.Conclusions. The results obtained prove that compounds 1-3 may have a potentially carcinogenic effect. However, this assumption requires further in-depth experimental studies.

A new method for the synthesis of azido-propargyloxy derivatives of 1,3,5-triazine has been developed utilizing the nitrosation of hydrazyno-1,3,5-triazines. New hydrazines (2-hydrazino-4,6-bis(propargyloxy)-1,3,5-triazine and 2,4-dihydrazino-6-propargyloxy-1,3,5-triazine) were synthesized and characterized via FTIR, NMR spectroscopy and elemental analysis. The hyperbranched polymers with azide (diazide monomer) and propargyloxy terminal groups were obtained via the azide-alkyne polycycloaddition reaction of diazide and monoazide AB2-type monomers. The antibacterial activity against Escherichia coli bacteria of 2,4,6-trispropargyloxy-1,3,5-triazine, 2-azido-4,6-bispropargyloxy-1,3,5-triazine, and 2,4-diazido-6-propargyloxy-1,3,5-triazine and their hyperbranched polymers was studied. Only 2,4-diazido-6-propargyloxy-1,3,5-triazine has weak antibacterial activity in comparison with ampicillin. The cytotoxicity of these compounds against M-HeLa, FetMSC, and Vero cell lines was also studied. 2,4,6-trispropargyloxy-1,3,5-triazine does not show any cytotoxic effect (IC50 ≥ 280 µM). It was shown that the presence of an azide group in the compound directly affects the cytotoxic effect. Hyperbranched polymers have a less cytotoxic effect against M-HeLa (IC50 > 100) in comparison with monomers (IC50 = 90–99 µM). This makes it possible to use these polymers as the basis for biocompatible materials in biomedical applications. [ABSTRACT FROM AUTHOR]

Scientific relevance. To date, no specific antivirals have been approved to treat and prevent Chikungunya fever, its complications, and sequelae. Therefore, the development of therapeutic and preventive medicinal products against Chikungunya virus (CHIKV), including interferon inducers, is gaining relevance.Aim. The authors aimed to study the effectiveness of prophylactic administration of an interferon inducer against CHIKV in an in vitro model.Materials and methods. The study used two cell lines (Vero and А549), a CHIKV strain (Nika2021), and an interferon-inducing medicinal product (double-stranded RNA sodium salt) at two doses (250 μg/mL and 500 μg/mL) administered at two schedules: Prevention (4 h prior to the virus challenge) and Emergency Prevention (at the time of the virus challenge). The authors determined the CHIKV titre by its cytopathogenic effect, the CHIKV RNA content by the cycle threshold value in real-time reverse-transcription polymerase chain reaction, and the concentration of cytokines using the enzyme immunoassay method. The study monitored the changes in CHIKV biological activity, CHIKV RNA levels, and the production of interferon-alpha (IFN-α), interferon-gamma (IFN-γ), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α) in cells over time. The statistical analysis of the resulting data used Microsoft Office Excel 2016 and StatTech.Results. The medicinal product at doses of 250 μg/mL and 500 μg/mL stimulated the production of both IFN-α and IFN-γ (IFN-α to a greater extent than IFN-γ) in both cell lines (in A549 to a greater extent than in Vero). The changes in CHIKV RNA levels with time corresponded to those of the virus titre. In general, CHIKV RNA levels in Vero cells were significantly higher than those in A549 cells (р

Scientific relevance. Tick-borne encephalitis is one of the most significant anthropozoonoses for public health in the Russian Federation. Wide vaccination coverage is required to control this zoonotic infection. However, large-scale immunisation is not feasible without developing novel vaccines with improved safety and efficacy profiles, such as vaccines based on the continuous Vero cell line.Aim. This extended study aimed to investigate the key quality attributes of VeroKSEN, a new tick-borne encephalitis vaccine that was obtained using Vero cells.Materials and methods. The authors implemented current approaches to vaccine development, including propagation of tick-borne encephalitis strain 205 (Far Eastern subtype) in Vero cells, fine purification of the viral antigen by size-exclusion chromatography on polymeric resins, and introduction of additional quality control methods. For VeroKSEN production, the authors used the method protected by utility patent No. 2678431. Quality control of the finished product was performed according to the State Pharmacopoeia of the Russian Federation. The laboratory study of the vaccine and its intermediates in Balb/с mice used novel control methods developed by the authors. Additional methods included polyacrylamide gel electrophoresis, enzyme immunoassay, immunoblotting with total antibodies to tick-borne encephalitis virus and monoclonal antibodies to glycoprotein E, reverse transcription–polymerase chain reaction, and high-performance liquid chromatography. The extended potency study used test strains of different subtypes, including Absettarov (Western), Sofjin (Far Eastern), and Korzukhin (Siberian). The authors studied the infectious activity of tick-borne encephalitis virus in outbred mice using the pharmacopoeial titration method. Statistical analysis of the study results involved calculating the arithmetic mean and the root-mean-square deviation.Results. The authors studied the key quality attributes of VeroKSEN and its intermediates. Selected inactivation conditions reduced the infectious activity of the viral harvest to an undetectable level within 24 h, while its antigenic activity remained approximately 100% of the baseline. The fine purification stages and the methods and techniques developed by the authors provided a whole-virion fraction with a purity of up to 98.2% and removed process-related impurities (residual host-cell DNA, bovine serum albumin, formaldehyde, and bacterial endotoxins) to the levels required by national legislation. The stability study demonstrated that the vaccine met the requirements for up to 36 months.Conclusions. The study showed the high potency of the new vaccine in terms of protection against the Western, Siberian, and Far Eastern subtypes of tick-borne encephalitis, with minimum immunising doses (MID50) of 0.007, 0.00125, and 0.00093 mL, respectively.

Introduction. The requirements of modern clinical guidelines for the treatment of respiratory viral infections suggest the possibility of identifying a specific viral pathogen. In this regard, the search for drugs with selective activity against respiratory syncytial virus and parainfluenza virus is relevant. The purpose of the work is to study the antiviral activity of the drug Cytovir®-3 in vitro in relation to the cytopathogenic effect of respiratory viruses (parainfluenza virus and respiratory syncytial virus). Material and methods. The antiviral effect of Cytovir®-3 in comparison with Umifenovir against parainfluenza virus and respiratory syncytial virus was studied on Vero cell culture. Drugs were administered 1 hour before (prophylactic regimen) and 1 hour after (treatment regimen) infection of the cell culture with respiratory viruses. The working range of concentrations of the studied drugs was calculated based on the values of 50% cytotoxic concentration calculated based on the results of a quantitative microtetrazole test. Results and discussion. The drug Cytovir®-3 in two schemes of application (therapeutic or prophylactic) showed its antiviral efficacy in vitro against respiratory syncytial virus and parainfluenza virus due in the non-toxic range (0–794 µg/ml). At the same time, the suppressive effect of the drug Cytovir®-3 against the parainfluenza virus begins at lower concentrations of the drug with its early (preventive) introduction into cell culture (250 mcg/ml). Conclusion. Antiviral activity of Cytovir®-3 has been proven in vitro against parainfluenza virus and respiratory syncytial virus. At the same time, in all series of experiments, Cytovir®-3 had a higher index of selectivity of antiviral action than that of the comparison drug Umifenovir.

The severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) pandemic highlighted the importance of vaccine innovation in public health. Hundreds of vaccines built on numerous technology platforms have been rapidly developed against SARS-CoV-2 since 2020. Like all vaccine platforms, an important bottleneck to viral-vectored vaccine development is manufacturing. Here, we describe a scalable manufacturing protocol for replication-competent SARS-CoV-2 Spike-pseudotyped vesicular stomatitis virus (S-VSV)–vectored vaccines using Vero cells grown on microcarriers in a stirred-tank bioreactor. Using Cytodex 1 microcarriers over 6 days of fed-batch culture, Vero cells grew to a density of 3.95 ± 0.42 ×106 cells/mL in 1-L stirred-tank bioreactors. Ancestral strain S-VSV reached a peak titer of 2.05 ± 0.58 ×108 plaque-forming units (PFUs)/mL at 3 days postinfection. When compared to growth in plate-based cultures, this was a 29-fold increase in virus production, meaning a 1-L bioreactor produces the same amount of virus as 1,284 plates of 15 cm. In addition, the omicron BA.1 S-VSV reached a peak titer of 5.58 ± 0.35 × 106 PFU/mL. Quality control testing showed plate- and bioreactor-produced S-VSV had similar particle-to-PFU ratios and elicited comparable levels of neutralizing antibodies in immunized hamsters. This method should enhance preclinical and clinical development of pseudotyped VSV-vectored vaccines in future pandemics.

An increase in cases of chikungunya fever is reported in the Caribbean, Central and South America, and Southeast Asia. As there is no specific treatment for this disease and the only available treatment is symptomatic, it is very relevant to develop vaccines against chikungunya fever. To develop an inactivated whole-virion vaccine against the disease, it is important to choose a susceptible cell culture that both provides high virus yields and is used for vaccine production.The aim of the study was to evaluate the susceptibility of multiple cell lines to Chikungunya virus infection and to select the monolayer culture method with the highest virus accumulation and yield.Materials and methods. The study used the CHIKV_Nic strain of the Chikungunya virus and cell lines C6/36 (for virus titration), CEF, MRC-5, Vero, and 4647. While choosing the culture method, the authors used culture flasks, a cell factory, and roller bottles. The authors determined the susceptibility of the cell lines to viral infection by the degree of accumulation of the infectious agent in the culture fluid. The results of virus titration were calculated on day 5 on the basis of a pronounced viral cytopathic effect.Results. The Vero and 4647 cell lines demonstrated the highest susceptibility to infection and virus concentrations in the culture fluid. The СEF and MRC-5 cell lines accumulated the virus at lower concentrations. The maximum virus titres (7.10–7.75 log10 TCID50/mL) were observed in the culture fluid 48 h after infection. The optimal multiplicity of infection (MOI) ranged between 0.001 and 0.0001 MOI/cell. At 0.0001 MOI/cell, the virus accumulated in the Vero cells cultured in roller bottles on day 2, with the maximum virus titre being 8.6±0.2 log10 TCID50/mL.Conclusions. Vero cells meet the safety and stability requirements set for the production of chikungunya vaccines. The study determined the minimum MOI of the Chikungunya virus for cell culture. The roller bottle culture method provides the highest cell culture yield and the highest titre of the virus in the culture fluid.

Objective. To study the antibacterial activity of lysostaphin against staphylococci various species, as well as its effect on the viability of Vero cells. Materials and Methods. Lysostaphin was obtained by genetic engineering. Purification of the protein was carried out on SP-sepharose, the purity was determined by electrophoresis in PAGE by Lamley. The susceptibility to lysostaphin of 9 species 175 strains of staphylococci was studied. Identification was performed by MALDI-TOF MS, antibiotic susceptibility by EUCAST (v. 11.0). The MIC of lysostaphin was studied by the method of serial dilutions with concentrations between 0.015 and 512 mg/l. For 72 hours, the viability of Vero cells with lysostaphin at concentrations of 0.5-32.0 mg/l was determined by the MTT method with counting of living cells according to their growth curve. The results were analyzed by ANOVA followed by Dunnett's test. Results. A kinetic study of S. aureus growth in the presence of revealed an inhibitory effect of endopeptidase (MIC 0.06 mg/l). Lysostaphin was characterized by pronounced activity against clinical methicillinsensitive S. aureus. The MIC ranged between 0.03 and 0.5 mg/l and the MIC50/90 was 0.125/0.5 mg/l. For methicillin-resistant S. aureus MIC50/90 0.25/0.5 mg/l. The MIC50 for MRSE was 2 times higher than for MSSE – 1 mg/l. The maximum MIC value was determined against isolates of S. warneri and S. hominis – 64 mg/l, the lowest for S. saprophyticus – 0.5 mg/l. MIC50 of lysostaphin against MRSA was 4 times lower than that of vancomycin, MIC90 was 3 times lower. Differences in viable cells depending on the concentration of lysostaphin were not found. Conclusions. Significant activity of lysostaphin against staphylococci was revealed, which is several times higher than vancomycin against MRSA. Lysostaphin was also effective against methicillin-resistant S. aureus. The high anti-staphylococcal activity and cytocompatibility of lysostaphin are promising for its further study in the prevention and treatment of staphylococcal orthopedic infections.

The COVID-19 virus has caused a global emergency and has attracted the attention of healthcare professionals and the public around the world. The significant increase in the number of new cases of infection with this virus demonstrates the relevance of the search for drugs that are effective against this pathogen. The aim of this work was to evaluate the antiviral efficacy of Mefloquin® against COVID-19. The antiviral efficacy of Mefloquin® against the new pandemic virus SARS-CoV-2 was studied in in vitro experiments in Vero C1008 cell culture and in vivo on Syrian golden hamsters. The results of the study revealed that the drug Mefloquine® at a concentration of 2.0 µg ml-1, when applied after infection of cells, suppresses the reproduction of the SARS-CoV-2 virus by 1.7–1.9 lg, the inhibition rate is about 99%. When using Mefloquine, pathological changes in the lung tissue were less pronounced than in the control group. 6 days after infection, it was shown that when using Mefloquine, there was a statistically significant decrease in viral load in the lungs of infected Syrian golden hamsters, with an inhibition rate of 95.5%.

The purpose of this research was to analyze polymer materials based on mechanical properties and geometrical parameters, such as the smallest material deviations and the best printing texture after three-dimensional (3D) printing in two methods of Material Jetting technology: PolyJet and MultiJet. This study covers checks for Vero Plus, Rigur, Durus, ABS, and VisiJet M2R-WT materials. Thirty flat specimens were printed both for 0 and 90 raster orientations. Specimen scans were superimposed on the 3D model from CAD software. Each of them was tested, paying attention to the accuracy and the layer thickness effect of printed components. Then, all specimens were subjected to tensile tests. The obtained data—Young's modulus and Poisson's ratio—were compared using statistical methods, focusing on the two most important parameters: the isotropy of the printed material in two directions and the characteristics close to linear. It was found that unitary surface deviation with general dimensional accuracy equal to ±0.1 mm was the common feature of printed models. Some small areas had lower accuracy depending on the material and printer device. Rigur material obtained the highest mechanical properties. Dimensional accuracy in Material Jetting technology as a function of layer parameters such as layer thickness and raster orientation was checked. The materials were checked in terms of relative isotropy and linearity. Additionally, similarities and differences between PolyJet and MultiJet methods were covered. [ABSTRACT FROM AUTHOR]

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of Coronavirus Disease 2019 (COVID-19), poses a serious global public health threat for which there is currently no satisfactory treatment. This study examines the efficacy of Biobran/MGN-3 against SARS-CoV-2. Biobran is an arabinoxylan rice bran that has been shown to significantly inhibit the related influenza virus in geriatric subjects. Here, Biobran's anti-SARS-CoV-2 activity was assessed using MTT and plaque reduction assays, RT-PCR, ELISA techniques, and measurements of SARS-CoV-2-related gene expression and protein levels. For Vero E6 cells infected with SARS-CoV-2, Biobran reduced the viral load by 91.9% at a dose of 100 μg/mL, it reduced viral counts (PFU/mL) by 90.6% at 50 μg/mL, and it exhibited a significant selectivity index (EC50/IC50) of 22.5. In addition, Biobran at 10 μg/mL inhibited papain-like proteinase (PLpro) by 87% and ACE2 SARS-CoV-2 S-protein RBD by 90.5%, and it significantly suppressed SARS-CoV-2 gene expression, down-regulating E-gene and RdRp gene expression by 93% each at a dose of 50 μg/mL and inhibiting the E-protein by 91.3%. An in silico docking study was also performed to examine the protein–protein interaction (PPI) between SARS-CoV-2 RBD and DC-SIGN as well as between serine carboxypeptidase and papain-like protease PLpro. Serine carboxypeptidase, an active ingredient in Biobran, was found to interfere with the binding of SARS-CoV-2 to its receptor DC-SIGN on Vero cells, thus preventing the cell entry of SARS-CoV-2. In addition, it impairs the viral replication cycle by binding to PLpro. We conclude that Biobran possesses potent antiviral activity against SARS-CoV-2 in vitro and suggest that Biobran may be able to prevent SARS-CoV-2 infection. This warrants further investigation in clinical trials. [ABSTRACT FROM AUTHOR]

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Copyright of Eurasian Journal of Applied Biotechnology is the property of National Center for Biotechnology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Cancer is one of the most serious diseases facing humanity, so we urgently need to find a cure which is rarely harmful to the patient as much as possible. It has been approved that arginine deiminase (ADI) can hydrolyze the plasma arginine to citrulline. This hydrolysis activity and reduction in the amount of inter cellular argenine suppress lipopolysaccharide-induced nitric oxide (NO) synthesis. On the other hand arginine depletion, arresting the cell cycle at G1 phase, Therefore, ADI has considered as a powerful anticancer agents. The aim of the current study was to investigate the lethal effects of ADI purified from Lactobacillus plantarum p5 strain on murine mammary adenocarcinoma and vero cell lines. Anti-proliferative activity of ADI against murine mammary adenocarcinoma )AMN3) cell line was evaluated after different incubation time (3, 6, 24, 48 and 72 h) of exposure to 1 µg/mL of ADI compared with Vero (none cancer cell line ) transformed cell line with same conditions. Autophagy process in cancer cells was recognized after three hours of incubation with ADI which was clearly observed in AMN3 cell line under inverted microscope. First stages of programmed cell death (apoptosis) pathway was only observed in AMN3 cells after 24 h of incubation with ADI and this process continued with the time until they reached to last stages of apoptosis after 72 h of incubation. The results of the current study showed that the AMN3 cell line was auxotrophic for arginine because it couldn't produce it in the presence of enzyme which had a robust activity to kill these cancer cells, but Vero none cancer cell line was survive in the presence of ADI because it had the ability to produce arginine.

In 2020–2021, the world was engulfed by the pandemic of a new coronavirus infection (COVID-19) caused by the SARS-CoV-2 virus. The low population coverage with vaccination against COVID-19 and the lack of herd immunity result in the need to find an effective and safe etiotropic treatment. Medicinal agents for treatment of COVID-19, approved while preparing this publication, have several limitations related to the conditions of their use and/or population category. In this situation, interferon-containing drugs widely used in Russia and the CIS for prevention and treatment of viral infectious diseases, i.e. ARVI and influenza, may hold promise. This study aims to confirm in vitro antiviral activity against SARS-CoV-2 for the preparation VIFERON® containing recombinant human interferon alpha-2b (IFNα-2b). Materials and methods. Vero CCL-81 cells were infected with hCoV-19/StPetersburg-RII3524VR4/2020 strain of SARS-CoV-2 at doses of 10 TCID50 or 100 TCID50 per well. The suppressive effect of IFN-2b, extracted from VIFERON® in dosage form of rectal suppositories, was evaluated by qRT-PCR at 24 h and 48 h after the infection of cells in two schemes, simulating preventive (24 h before infection) and therapeutic (2 h after infection) use of drugs. Results. IFNα-2b at concentrations of 800, 400, 200, 100 and 50 IU/ml, extracted from rectal suppositories of VIFERON®, showed high biological activity, displayed as inhibition of SARS-CoV-2 strain replication in both infectious doses evaluated either at 24 h or at 48 h after cell infection. The “preventive” vs. “therapeutic” scheme was found to be more effective. In the “preventive” scheme the virus titre decreased by more than 3 lg TCID50 at 24 hours post-infection and by 5–6 lg TCID50 at 48 hours post-infection after administration of 800 IU/ml IFNα-2b. Conclusion. The study results evidence that VIFERON® in dosage form of rectal suppositories may be promising for prevention and treatment of new coronavirus infection in clinical practice.

N-acetylneuraminic acid linked to galactose by α2,6 and α2,3 linkages (Siaα2,6 and Siaα2,3) is expressed on glycoconjugates of animal tissues, where it performs multiple biological functions. In addition, these types of sialic acid residues are the main targets for the binding and entry of influenza viruses. Here we used fluorochrome-conjugated Sambuccus nigra, Maackia amurensis, and peanut lectins for the simultaneous detection of Siaα2,3 and Siaα2,6 and galactosyl residues by two-color flow cytometry on A549 cells, a human pneumocyte cell line used for in vitro studies of the infection by influenza viruses, as well as on Vero and MDCK cell lines. The dexamethasone (DEX) glucocorticoid (GC), a widely used anti-inflammatory compound, completely abrogated the expression of Siaα2,3 in A549 cells and decreased its expression in Vero and MDCK cells; in contrast, the expression of Siaα2,6 was increased in the three cell lines. These observations indicate that DEX can be used for the study of the mechanism of sialylation of cell membrane molecules. Importantly, DEX may change the tropism of avian and human/pig influenza viruses and other infectious agents to animal and human epithelial cells. [ABSTRACT FROM AUTHOR]

Several cancers are induced by microbial infections or chronic inflammation. Ptaeroxylon obliquum is traditionally used to treat various infections characterized by inflammation. The in vitro antiproliferative and antioxidant activity of P. obliquum leaf extracts, fractions and isolated compounds were determined. Antiproliferative activity was assessed against normal Vero cells, and several cancerous human cells, including human breast cancer (MCF-7), hepatocarcinoma (HepG2), lung adenocarcinoma (A549) and human cervical cancer cells (HeLa) using a colorimetric tetrazolium bromide assay. Radical scavenging activity was tested using the 2,2-diphenyl-1-instrpicrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. Obliquumol, O-methylalloptaeroxylin and a mixture of lupeol and β-amyrin were isolated from the chloroform fraction using silica gel open column chromatography. Acetone extracts were toxic to HepG2 cells with IC50 values from 8 to 200 µg/mL but were less toxic to other cells with selectivity index as high as 14. Aqueous extracts and fractions were non-toxic at concentrations tested against all the cell lines (IC50 > 100 µg/mL). Isolated compounds had IC50 values ranging from 52 to 539 µg/mL and 189 to 247 µg/mL against HepG2 and HeLa cells, respectively. Light microscopy showing changes in HepG2 and HeLa cell morphology supported the cytotoxicity of the acetone extracts. Water extracts scavenged ABTS and DPPH radicals with IC50 values as low as 29.06 µg/mL and 43.4 µg/mL. P. obliquum extracts may be useful as sources of anticancer therapy, as they have selective cytotoxicity against cancer cell lines. [ABSTRACT FROM AUTHOR]

Dengue virus (DENV), a common and prevalent mosquito-borne endemic disease, is caused by four serotypes (DENV-1-4) and has spread rapidly on a global scale over the past decade. A crucial step in the development of antiviral therapeutics requires the utilization of in vitro cell-based techniques, such as plaque assays and focus-forming assays (FFA) for virus quantification. Vero cells have been widely used for FFA and plaque assay; however, there are instances when their efficacy and efficiency in the detection of certain clinical DENV isolates are low. Here, we showed that BHK-21 cells are more sensitive than Vero cells in the detection of all DENV-1-4 plaques and foci. In addition, we developed an improved FFA protocol for the quantification of all four DENV serotypes. Using a pan-flavivirus envelope (E) antibody, we reduce the possibility of false positives by defining a focus to consist of a minimum of eight infected cells. We outlined a protocol using the Operetta® high-content imaging system to automate the digital capture of these infected cells. A pipeline was also designed using the CellProfilerTM automated image analysis software to detect these foci. We then compare the results of the improved FFA with plaque assay. Notably, the improved FFA detected clear foci of the DENV-4 strain that does not form distinct plaques. We subsequently demonstrated the potential application of the improved FFA protocol in antiviral testing, utilizing a nucleoside inhibitor of DENV, NITD008 as a control. The protocol is amenable to a diverse array of applications, including high-throughput compound screening (HTS). Key features • An improved focus-forming assay that has the potential to quantify the DENV-4 strain, which was previously hard to plaque. • Improvements have been made to reduce the possibility of false positives. • Improved workflow using automated digital imaging process and counting of foci. • Applicable to antiviral compounds screening and is amenable to high-throughput screening., Competing Interests: Competing interestsAuthors declare no competing interests., (©Copyright : © 2024 The Authors; This is an open access article under the CC BY-NC license.)

In China, Japan, and Korea, Panax ginseng has been used in traditional medicine for thousands of years. Panax is a plant used as a general tonic or adaptogen for chronically ill patients. The current study evaluated the cytotoxicity of Panax ginseng extract (PGE). Different cell lines (HCT-116, LNCaP, and normal cell line VERO) were treated with different inhibitory agentsat different concentrations (1000, 500, 250, 125, 62.5, and 31.25 µg/ml) as follows: G1 (Methanol Panax ginseng extract, PGE), G2 (Doxorubicin, DOX), and G3 (Methanol Panax ginseng extract +DOX, PDD). Each inhibitory agent group was used to treat the cancerous cell lines HCT-116, LNCaP, and normal cell line (VERO) to obtain IC50% by MTT assay. The inhibitory ability of the 1000 μg/ml PGE was significantly increased in all the three-cell lines compared with other concentrations. The recorded data revealed that the inhibition ability of PGE and Doxorubicin towards the HCT-116 cell line significantly increased compared with the other cell lines. The interaction between different PGE concentrations and cell lines showed that the 1000 μg/ml PEG had the highest inhibitory effects on HCT-116 compared with other combinations. The interaction between different DOX concentrations and different types of cell lines showed that the 1000 μg/ml DOX had the highest inhibitory effects on LNCap compared with other combinations. The PGD inhibition ability reflected a significantly higher difference toward the HCT-116 cell line as compared with other cell lines. IC50% is the concentrations (µg/ml) to kill 50% of cell line. It was calculated by MTT assay for three cell lines: HCT-116, LNCaP, and VERO. The rate of effectiveness of the inhibitory factors (PGE, DOX, and PGD) showed highly significant differences toward the cell line HCT-116 compared to the other cell lines. This indicates the safety of the PGE compound and its low toxicity toward normal cells, quite the opposite of cancer cells as compared to the common drug DOX and combined PGD (PGE+DOX). PGD combined with DOX (PGE + DOX) showed antagonistic results toward the HCT116, LNCaP, and VERO cell lines, while UDE combined with DOX (UDE+DOX) showed synergistic activity.

On account of the COVID-19 pandemic, the global pharmaceutical industry has achieved impressive results in the development and introduction of various types of vaccines causing the formation of acquired immunity against the SARS-CoV-2 coronavirus into clinical practice. However, none of them currently show the declared one hundred percent guarantee of protection. In the case of the COVID-19 disease, patients with concomitant pathologies are the most vulnerable to the occurrence of severe complications. The aerosol route of transmission of SARS-CoV-2 contributes to the emergence of outbreaks of the new coronavirus infection in crowded places and closed rooms with poor ventilation. In this regard, an urgent problem is the search for drugs with local antiviral activity, which, together with restrictive measures and mask wearing policy, can potentially reduce the likelihood of contracting coronavirus. In this experimental in vitro study on Vero CCL81 cell culture (ATCC), the local antiviral activity of the drug Thymogen® spray against the SARS-CoV-2 virus was studied in comparison with the antiseptic Miramistin® solution. As a result of the experiment, no toxic effects on Vero cells were detected in the drugs in the studied concentrations. In a series of experiments, Thymogen® spray showed local antiviral activity against SARS-CoV-2 when the virus titer was 5,2 lg TCID50. Therefore, the drug Thymogen® dosed nasal spray has high potential as a topical drug for prevention and treatment of COVID-19 disease, which requires additional confirmation in relevant clinical studies.

Introduction. The COVID-19 pandemic has stimulated the search for drugs with specific antiviral activity against the new pathogenic strain of the SARS-CoV-2 coronavirus. First of all, scientific search was aimed at studying drugs with already proven efficacy against influenza and ARVI. The aim of this work was to study the antiviral activity of Cytovir®-3 in vitro in relation to the cytopathogenic effect of the SARS-CoV-2 virus. Material and methods. The antiviral activity of the drug Cytovir®-3 against the SARS-CoV-2 virus was studied in experimental models in vitro on Vero CCL81 cell culture (ATCC). The maximum tolerated concentration and the 50% cytotoxic dose were determined using a quantitative microculture tetrazolium test assay to calculate the working range of the concentrations of the test drug. Results and discussion. As a result of the study, it was shown that the greatest activity of the drug was manifested when it was added to the cells 24 hours before and 1 hour and 24 hours after viral infection, the inhibition level reached 53% (>IC50) at the drug concentrations of 105, 55, and 85 µg/ml, respectively. Cytovir®-3 suppressed the viral activity of SARS-CoV-2 in the dose range from 10 µg/ml to 105 µg/ml under the indicated infection conditions. It was found that the drug did not exhibit cytotoxic effects on the Vero cell culture in the range of antiviral doses. Conclusion. The antiviral activity of Cytovir®-3 against the SARS-CoV-2 virus has been proven due to the achievement of IC50, which is below the maximum tolerated dose of 149 µg/ml.

Introduction. The pandemic spread of a new coronavirus infection, COVID-19, has caused a global emergency and attracted the attention of public health professionals and the population of all countries. A significant increase in the number of new cases of SARS-CoV-2 infection demonstrates the urgency of finding drugs effective against this pathogen. The aim of this work was to evaluate the in vitro antiviral efficacy of human recombinant alpha-2b interferon (IFN-α2b) against SARS-CoV-2 virus. Material and methods. The experiments had been carried out on Vero Cl008, the continuous line of African green monkey (Chlorocebus sabaeus) kidney cells. The effectiveness of the drugs was assessed by the suppression of viral reproduction in vitro. The biological activity was determined using titration of a virus-containing suspension in a Vero Cl008 cell culture by the formation of negative colonies. Results. The antiviral efficacy of the IFN-α2b-based medications, which have a high safety profile and proven efficacy in the prevention and treatment of influenza and acute respiratory viral infections (ARVI), has been studied against the new pandemic SARS-CoV-2 virus in vitro experiments in Vero C1008 cell culture. IFN-α2b effectively inhibits the reproduction of the virus when applied both 24 hrs before and 2 hrs after infection. In the IFN-α2b concentration range 102–106 IU/ml a complete suppression of the reproduction of the SARS-CoV-2 virus had been demonstrated. Discussion. IFN-α2b demonstrated in vitro high antiviral activity against SARS-CoV-2. In addition, the substance has a high chemotherapeutic index (1000). Conclusion. Medications for intranasal use based on IFN-α2b have high antiviral activity and are promising drugs for in vivo study in terms of prevention and treatment of COVID-19.

Lung cancer ranks as the 2nd most common cancer globally. It's the most prevalent cancer in men and the 2nd most common in women. The prominent events in EGFR-mutated non-small-cell lung cancer (NSCLC) include the emergence of the L858R mutation within EGFR exon 21. Despite the promising efficacy of EGFR inhibitors in managing lung cancer, the development of acquired resistance poses a significant hurdle. In the current investigation, we focused on the screening of two phytochemicals, namely Dehydrocostus lactone and Mokkolactone, derived from the Saussurea lappa plant, as potential inhibitors targeting EGFR L858R mutant lung cancer. The chloroform and ethanol extract of the plant demonstrated anti-proliferative activity through the Resazurin chemosensitivity assay, exhibiting an IC50 value of 37.90 ± 0.29 µg/ml with selectivity index 2.4. Through a GC–MS study, we identified 11 phytochemicals for further insilico analysis. These compounds underwent ADMET assessment followed by drug likeliness analysis before being subjected to molecular docking against EGFR L858R, identified through protein–protein interaction network analysis. All phytochemicals exhibited binding energy scores ranging from −6.9 to −8.1 kcal/mol. Dehydrocostus lactone and Mokkolactone were specifically identified for their binding profile. Findings from 100 ns molecular dynamics simulations demonstrated their enhanced stability compared to the reference ligand DJK. This was evident in the root mean square deviation (RMSD) values, ranging from 0.23 ± 0.01 nm to 0.30 ± 0.05 nm, the radius of gyration values, from 1.71 ± 0.01 nm to 1.72 ± 0.01 nm, and the solvent accessible surface area values, from 155.39 ± 2.40 nm2 to 159.32 ± 2.14 nm2. Additionally, favourable characteristics were observed in terms of hydrogen bonding, principal component analysis, and free energy landscape analysis. Examination of their electronic structure via density functional theory revealed efficient properties, with the highest occupied molecular orbital-least unoccupied molecular orbital energy gap values ranging from −3.984 eV to −6.547 eV. Further, in vivo analysis is required to gain a more comprehensive understanding and efficacy of these identified phytochemicals against lung cancer. [ABSTRACT FROM AUTHOR]

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Cancer is one of the most serious diseases facing humanity; accordingly, it is urgent to find a cure that is rarely harmful to the patient as much as possible. It has been approved that arginine deiminase (ADI) can hydrolyze the plasma arginine to citrulline. This hydrolysis activity and reduction in the amount of intercellular arginine suppress lipopolysaccharide-induced nitric oxide synthesis. On the other hand, arginine depletion arrests the cell cycle at the G1 phase; therefore, ADI has been considered a powerful anticancer agent. The current study aimed to investigate the lethal effects of ADI purified from the Lactobacillus plantarum p5 strain on murine mammary adenocarcinoma and Vero cell lines. Anti-proliferative activity of ADI against murine mammary adenocarcinoma) AMN3) cell line was evaluated after different incubation times (3, 6, 24, 48, and 72 h) of exposure to 1 μg/mL of ADI, compared to Vero (non-cancer cell line) transformed cell line with same conditions. The autophagy process in cancer cells was recognized after three hours of incubation with ADI which was clearly observed in the AMN3 cell line under an inverted microscope. The first stages of the programmed cell death (apoptosis) pathway were only observed in AMN3 cells after 24 h of incubation with ADI, and this process continued with the time until they reached the last stages of apoptosis after 72 h of incubation. The results of the current study showed that the AMN3 cell line was auxotrophic for arginine because it could not produce it in the presence of enzyme which had a robust activity to kill these cancer cells; however, Vero non-cancer cell line survived in the presence of ADI because it had the ability to produce arginine. [ABSTRACT FROM AUTHOR]

The antiviral activity of the drug Kagocel®, which has a high safety profile and proven efficacy for the prevention and treatment of influenza and AVRI as an interferon inducer, was studied against a new pandemic strain of SARS-CoV-2 in vitro in Vero C1008 cell culture. The results of the study revealed that at the addition of the substance Kagocel® at the concentration of 5000 pg/ml into the cell culture 1 h before the virus infection and 1 h after, there was 100% inhibition of the cytopathic activity of the virus. It was also found that Kagocel® at the dose of 5000 pg/ml effectively suppressed the reproduction of the SARS-CoV-2 virus, vari­ant B, in Vero C1008 cell culture by 1.75 lg, the inhibition coefficient was 97.83 %.

N-acetylneuraminic acid linked to galactose by α2,6 and α2,3 linkages (Siaα2,6 and Siaα2,3) is expressed on glycoconjugates of animal tissues, where it performs multiple biological functions. In addition, these types of sialic acid residues are the main targets for the binding and entry of influenza viruses. Here we used fluorochrome-conjugated Sambuccus nigra, Maackia amurensis, and peanut lectins for the simultaneous detection of Siaα2,3 and Siaα2,6 and galactosyl residues by two-color flow cytometry on A549 cells, a human pneumocyte cell line used for in vitro studies of the infection by influenza viruses, as well as on Vero and MDCK cell lines. The dexamethasone (DEX) glucocorticoid (GC), a widely used anti-inflammatory compound, completely abrogated the expression of Siaα2,3 in A549 cells and decreased its expression in Vero and MDCK cells; in contrast, the expression of Siaα2,6 was increased in the three cell lines. These observations indicate that DEX can be used for the study of the mechanism of sialylation of cell membrane molecules. Importantly, DEX may change the tropism of avian and human/pig influenza viruses and other infectious agents to animal and human epithelial cells.

Several cancers are induced by microbial infections or chronic inflammation. Ptaeroxylon obliquum is traditionally used to treat various infections characterized by inflammation. The in vitro antiproliferative and antioxidant activity of P. obliquum leaf extracts, fractions and isolated compounds were determined. Antiproliferative activity was assessed against normal Vero cells, and several cancerous human cells, including human breast cancer (MCF-7), hepatocarcinoma (HepG2), lung adenocarcinoma (A549) and human cervical cancer cells (HeLa) using a colorimetric tetrazolium bromide assay. Radical scavenging activity was tested using the 2,2-diphenyl-1-instrpicrylhydrazyl (DPPH) and 2,2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. Obliquumol, O-methylalloptaeroxylin and a mixture of lupeol and β-amyrin were isolated from the chloroform fraction using silica gel open column chromatography. Acetone extracts were toxic to HepG2 cells with IC50 values from 8 to 200 µg/mL but were less toxic to other cells with selectivity index as high as 14. Aqueous extracts and fractions were non-toxic at concentrations tested against all the cell lines (IC50 > 100 µg/mL). Isolated compounds had IC50 values ranging from 52 to 539 µg/mL and 189 to 247 µg/mL against HepG2 and HeLa cells, respectively. Light microscopy showing changes in HepG2 and HeLa cell morphology supported the cytotoxicity of the acetone extracts. Water extracts scavenged ABTS and DPPH radicals with IC50 values as low as 29.06 µg/mL and 43.4 µg/mL. P. obliquum extracts may be useful as sources of anticancer therapy, as they have selective cytotoxicity against cancer cell lines.

Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The available cell-adapted hepatitis A virus (HAV) strains show a very slow replication phenotype hampering the affordable production of antigen. A fast-growing strain characterized by the occurrence of mutations in the internal ribosome entry site (IRES), combined with changes in the codon composition has been selected in our laboratory. A characterization of the IRES activity of this fast-growing strain (HM175-HP; HP) vs. its parental strain (HM175; L0) was assessed in two cell substrates used in vaccine production (MRC-5 and Vero cells) compared with the FRhK-4 cell line in which its selection was performed. The HP-derived IRES was significantly more active than the L0-derived IRES in all cells tested and both IRES were more active in the FRhK-4 cells. The translation efficiency of the HP-derived IRES was also much higher than the L0-derived IRES, particularly, in genes with a HP codon usage background. These results correlated with a higher virus production in a shorter time for the HP strain compared to the L0 strain in any of the three cell lines tested, and of both strains in the FRhK-4 cells compared to Vero and MRC-5 cells. The addition of wortmannin resulted in the increase of infectious viruses and antigen in the supernatant of FRhK-4 infected cells, independently of the strain. Finally, the replication of both strains in a clone of FRhK-4 cells adapted to grow with synthetic sera was optimal and again the HP strain showed higher yields. [ABSTRACT FROM AUTHOR]

The present study discloses the identification of phenolic compounds in Moringa oleifera hot water extract (MOH) and the evaluation of its antioxidant activity on H2O2-induced oxidative stress in Vero cells. Upon analysis, MOH was found to contain phenolic compounds and indicated 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS+) radical scavenging with IC50 values of 102.52 and 122.55 µg/mL, respectively. The ferric reducing antioxidant power (FRAP) of MOH indicated a dose-dependent increase with a maximum absorbance at 125 μg/mL and the oxygen radical absorbance capacity (ORAC) of MOH was 1004.95 µmol TE/mg. Results showed that MOH dose-dependently reduced intracellular ROS generation in H2O2-stimulated Vero cells while increasing the cell viability. Fluorescence microscopy and flowcytometric analyses have supported the above findings. MOH markedly suppressed the H2O2-induced mitochondrial depolarization and apoptosis through suppression of the mitochondrial-mediated apoptosis pathway and activated the Nrf2/HO-1 signaling pathway by possibly involving H2O2 generation in cell media. Findings of western blot were supported by immunocytochemistry of Nrf2 nuclear translocation. Thus, MOH bioactivity would potentiate its applications in manufacturing functional food.

BACKGROUND AND OBJECTIVE: Studies have shown that plants of Family Cuscutaceae can affect the growth and development of cancer cells. It is important that the herbs used for non-cancerous cells do not have cytotoxic effects. The present study was conducted to evaluate the effects of hydroalcoholic extracts of Cuscuta europaea on the viability of breast cancer cells (MCF7) compared to non-cancerous kidney epithelial cells (Vero cell line). METHODS: In this experimental study, Vero cells and MCF7 cells were prepared from Pasteur Institute Cell Bank and divided into the control group and the groups treated with doses of 0.01, 0.1, 1 and 10 mg / ml. 48 hours after exposure, the cytotoxic effect of the extract was measured by MTT. FINDINGS: Compared with the control group, exposure to dose of 0.01, 0.1, 1 and 10 mg/ml of hydroalcoholic extract of Cuscuta europaea reduced the viability of MCF7 cells (54.47±0.35, 55±0.42, 44.62±0.38, and 4.64±0.73%, respectively) and Vero cells (58.42±0.45, 48.61±0.55, 43.14±0.23, and 5.35±0.27%, respectively) (p

Background: Trichomoniasis is a common urogenital disease in the world. The first line of treatment for Trichomoniasis is metronidazole. Drug resistance and side effects of metronidazole urge researchers to seek new medications. Curcumin is a yellow substance derived from turmeric and has different derivatives with anticancer and antioxidant effects. We evaluated the anti-trichomonas effects of curcumin and its derivatives. Materials and Methods: Curcumin 70%, curcumin 90%, Bisdemethoxy curcumin, Diacetyl curcumin, Vanadyl curcumin, Vanadyl diacetyl curcumin, Indium curcumin and Gallium curcumin were prepared. Different concentrations (50, 100, 200, 400, 600, 800 µg/ml) of curcumin prepared with glycerin and 106 Trichomonas vaginalis were added to each well and incubated at 37°C for 24 h. The in vitro toxicity of different extracts against Trichomonas vaginalis was evaluated through MTT assay. All tests were performed in triplicate and SPSS software was used for data analysis. Results: The IC50 was 450, 400, 441, 453.2, 427.3, 417.6, 441 and 449.1 µg/ml, respectively for curcumin 70%, curcumin 90%, Bisdemethoxy curcumin, Diacetyl curcumin, Vanadyl curcumin, Vanadyl diacetyl curcumin, Indium curcumin and Gallium curcumin against Trichomonas vaginalis. Cytotoxic effects of these compounds against vero cells were 33.1%, 19%, 21%, 20.3%, 17%, 21%, 25.3% and 16%, respectively. Conclusion: Curcumin 90% had the most activity against Trichomonas vaginalis and Galium curcumin had the least cytotoxic effect against vero cells. It appears curcumin lotion can be used topically to treat trichomoniasis.

Cancer treatment remains a serious challenge worldwide. Thus, finding novel antitumour agents is of great importance. In the present study, nine new benzenesulphonohydrazide derivatives (1-9) were synthesized, and the chemical structures of the obtained compounds were confirmed by spectral analysis methods, including IR, 1H nuclear magnetic resonance (NMR) and 13C NMR. Experimental lipophilicity values were established using reversed phase-high performance thin layer chromatography. The antiproliferative activity of the synthesized compounds was tested against three tumour cell lines (769-P, HepG2 and NCI-H2170) and one normal cell line (Vero). Among the newly developed molecules, compound 4 exhibited generally the highest cytotoxicity across all tumour cell lines, and it was highly selective. However, higher selectivity towards the tested cancer cell lines was observed using compound 2, when compared with compound 4, which also exhibited significant antiproliferative activity against these tumour cells. In 769-P cells, compounds 5 and 6 were the most selective among all tested compounds. Compound 5 exhibited high cytotoxicity with an estimated IC50 value of 1.94 µM. In the NCI-H2170 cell line, compound 7 was the most cytotoxic and the most selective. In brief, the combination of fluorine and bromine substituents at the phenyl ring showed the most promising results, exerting high cytotoxicity and selectivity towards cancer cells. The renal adenocarcinoma cell line (769-P) appeared to be the most sensitive to the anticancer properties of the novel benzenesulphonohydrazones. [ABSTRACT FROM AUTHOR]

We generated plasmid pools for the rapid preparation of candidate vaccine strains, which could grow in the Vero cells at low temperature. Firstly, we cloned in the pHW2000 plasmid each of the eight gene segments (PB2, PB1, PA, hemagglutinin [HA], neuraminidase [NA], NS, NP, M) of two master donor strains (MDS), respectively, A/Yunnan/1/2005Vca(H3N2) and B/Yunnan/2/2005Vca(By), which had Vca phenotype (cold‐adapted phenotype in Vero cells). Secondly, the similar operation was implemented with each of the HA, NA and NP segments of circulating strains with epidemic potential (parental strains). The virus rescue techniques were employed in this study, according to the homology rate of HA segments between MDS and parental strains. Then, we harvested amount of new Vca virus strains. By transmission electron microscope, it could observe new viruses' diameter and length were from 100 to 120 nm. Importantly, these reassortant viruses could get high‐yield production in Vero cells at 25℃ from the beginning to the fourth generation, which was significantly differ from their original parental viruses. Additional, these production 16 new Vca strains could maintain enough antibody binding capacity and attenuation phenotype, which consisted with their MDS. So these plasmid pools constructed by mount of different influenza A and B virus gene fragments could present desired working performance and provide convenience and realization for more Vca reassortant virus as candidate vaccine strain if needing. [ABSTRACT FROM AUTHOR]

Background: Trichomonas vaginalis protozoa is one of the causes of human vaginitis. The selective medicine for treatment is metronidazole that in addition to increasing drug resistance, it also has many side effects. Pergularia tomentosa is a dessert plant that has the anti-fungal and anti- Plasmodium effect and anti dermatophytic effect of Priploca aphylla is also more effective than common antifungal medicine such as griseofulvin. In this study, we hope to use of these plants to introduce a new medicine combination. Materials and Methods: Pergularia tomentosa and Priploca aphylla were cleaned and dried after collecting, then each of the plants were extracted by using water, methanol, dicholoromethane and n-hexane solvents and with solvents mentioned concentrations 50, 100, 200, 400, 600, 800 µg/ml were prepared by using the normal saline and glycerin. 106 Trichomonas vaginalis trophozoite was added in 100 µl, then add 100 ml of the plants extract of Pergularia tomentosa and Priploca aphylla were added and incubated for 24 h at 37°C. Measurement of mortality rate of this protozoan were evaluated by using the MTT test. The effect of the extracts was performed on vero cells and Phytochemical evaluation was used by using the technique (HPTLC) to determine the composition of the extract. All tests wererepeated 3 times. SPSS 16 software were used for data analysis. Results: Trichomonas lethal effect of aqueous, methanol, dichloromethane and n-hexane extracts of Pergularia tomentosa in 800µg/ml concentration were % 64, % 72.4, %95.2 and % 95.4 respectively. anti Trichomonas effect of Priploca aphylla and with same concentration were 70.8%, 67%, 93.4% and 93.2% respectively. Cytotoxic effect of methanol extracts of Pergularia tomentosa and Priploca aphylla in 800µg/ml concentration against vero cells obtained %50.3 and %63 respectivley. Phytochemical screening of both plants extracts showed that both have the ingredients alkaloid, flavonoid, terpenoid, saponin, sterol, phenol, iridoid, tannin, phenylpropanoid, and anthocyanines. Conclusion: results showed that the Pergularia tomentosa and Priploca aphylla, dichloromethane and n-hexane extracts had more potent effect against Trichomonas vaginalis than methanolic and water extracts in all concentrations. so, identification and evaluation of the mechanism of these ingredients against Trichomonas vaginalis is recommended.

A total of 407 samples were collected from the western region of Saudi Arabia. Soil samples as well as dead larvae of Spodoptera littoralis (cotton leaf worm) were collected and examined for the presence of Bacillus thuringiensis. The bacterium was isolated using an acetate-selective enrichment medium and plating. The isolates that were collected by the above-mentioned protocol were identified by using a combination of both microscopic examinations as well as the analysis of 16S rRNA genes through DNA sequencing of PCR products. A total of 22 confirmed isolates of Bacillus thuringiensis were identified for the purpose of this study. Collectively, these confirmed isolates make up 4.6% of all soil samples and 6.6% of all dead larvae samples that were collected. Although B. thuringiensis was not found to be abundant in soil habitats of The Makkah Province, our results suggest that the bacterium is the part of the indigenous microflora of the area that we explored in this study. Preliminary results show that the partially purified crystal protein of some of the isolates exhibits demonstrable activity against three human cancer cell lines: Hep G2 (human liver carcinoma cell line), HeLa cells (human uterine cervix cancer cell line), and Vero cells (lung cancer Cell line).

ABSTRACT The use of high-throughput sequencing (HTS) to identify viruses in biologicals differs from current molecular approaches, since its use enables an unbiased approach to detection without the need to design specific primers to preamplify target sequences. Its broad range of detection and analytical sensitivity make it an important tool to ensure that biologicals are free from adventitious viruses. Similar to other molecular methods, however, identification of viral sequences in cells by HTS does not prove viral infection, since this could reflect carryover of inert viral sequences from reagents or other sources or the presence of transcriptionally inactive cellular sequences. Due to the broad range of detection associated with HTS, the above can potentially be perceived as a drawback for the testing of pharmaceutical biological products using this method. In order to avoid the identification of inert viral sequences, we present a methodology based on metabolic RNA labeling and sequencing, which enables the specific identification of newly synthesized viral RNAs in infected cells, resulting in the ability to unambiguously distinguish active infection by DNA or RNA viruses from inert nucleic acids. In the present study, we report the ability to differentiate Vero cells acutely infected by a single-stranded positive-sense RNA virus (tick-borne encephalitis virus) from cells which have been in contact with nonreplicating virus particles. Additionally, we also found a laboratory contamination by the squirrel monkey retrovirus of our Vero cell line, which was derived from an Old World (African green) monkey, a type of contamination which until now has been identified only in cells derived from primates from the New World. IMPORTANCE The use of high-throughput sequencing (HTS) to identify viral contamination of biological products is extremely sensitive and provides a broad range of detection. Nevertheless, viral sequences identified can also be inert. Examples include contamination resulting from reagents or the presence of inactivated viruses in animal-derived components of the cell culture medium. We therefore developed a method that relies on the sequencing of newly synthesized RNAs, an unequivocal sign of the presence of a transcriptionally active virus. This improvement in the specificity of viral testing increases the acceptability of HTS as a standard test for cells used in manufacturing biologicals and in biotherapies.

Abstract INTRODUCTION: Insect cell cultures play an essential role in understanding arboviral replication. However, the replicative efficiency of some of these viruses such as dengue (DENV), yellow fever (YFV), and chikungunya (CHIKV) in a new cellular substrate (Lulo) and in the other two recognized cell lines has not been comparatively assessed. METHODS: Vero, C6/36, and Lulo cell lines were infected with DENV, YFV, and CHIKV. The viral progeny was quantified through plaque assays and quantitative reverse transcription-polymerase chain reaction, while for DENV2, the findings were confirmed by immunofluorescence antibody assay. RESULTS: The higher DENV2 titer (from multiplicity of infection 0.001) was obtained on day four post-infection in C6/36 and on day six in Vero cells, while the Lulo cell line was almost impossible to infect under the same conditions. However, C6/36 showed the highest values of viral RNA production compared to Vero cells, while the quantification of the viral RNA in Lulo cells showed high levels of viral genomes, which had no correlation to the infectious viral particles. CONCLUSIONS: C6/36 was the most efficient cell line in the alpha and flavivirus production, followed by Vero cells. Thus, Lulo cells may be a useful substrate to study the mechanisms by which cells evade viral replication.

The severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) pandemic highlighted the importance of vaccine innovation in public health. Hundreds of vaccines built on numerous technology platforms have been rapidly developed against SARS-CoV-2 since 2020. Like all vaccine platforms, an important bottleneck to viral-vectored vaccine development is manufacturing. Here, we describe a scalable manufacturing protocol for replication-competent SARS-CoV-2 Spike-pseudotyped vesicular stomatitis virus (S-VSV)-vectored vaccines using Vero cells grown on microcarriers in a stirred-tank bioreactor. Using Cytodex 1 microcarriers over 6 days of fed-batch culture, Vero cells grew to a density of 3.95 ± 0.42 ×10 6 cells/mL in 1-L stirred-tank bioreactors. Ancestral strain S-VSV reached a peak titer of 2.05 ± 0.58 ×10 8 plaque-forming units (PFUs)/mL at 3 days postinfection. When compared to growth in plate-based cultures, this was a 29-fold increase in virus production, meaning a 1-L bioreactor produces the same amount of virus as 1,284 plates of 15 cm. In addition, the omicron BA.1 S-VSV reached a peak titer of 5.58 ± 0.35 × 10 6 PFU/mL. Quality control testing showed plate- and bioreactor-produced S-VSV had similar particle-to-PFU ratios and elicited comparable levels of neutralizing antibodies in immunized hamsters. This method should enhance preclinical and clinical development of pseudotyped VSV-vectored vaccines in future pandemics., Competing Interests: The authors declare no competing interests., (© 2024 The Author(s).)

Essential oils (EO) possess a wide range of biological activities. However, their application in aqueous media is often limited due to their hydrophobicity and volatile character. This study was designed to prepare stable, water-dilutable microemulsions (ME) containing essential oils of citronella (Cymbopogon nardus (L.) Rendle), mint (Mentha x piperita L. 'Multimentha') and eucalyptus (Eucalyptus globulus Labill.) and to evaluate their in vitro antioxidant and cytotoxic properties. The comparison of cytotoxicity of EO solubilised in microemulsions and in dimethyl sulfoxide as well as the recovery of volatiles from cells culture medium over time was also performed. The clear ME were obtained in a range between 10% and 50% of aqueous phase for citronella EO and up to 60% of aqueous phase for mint and eucalyptus EO, in all ratios of Tween 80 to oil phase (from 5:1 to 9:1). Microemulsions of EO (EO/ME) showed higher antioxidant activity compared to EO. The increase in activity was 13.96%, 22.25% and 45.60% for eucalyptus, mint and citronella EO, respectively. The analysis of cytotoxicity profiles of EO/ME and EO/DMSO in Vero and HeLa cell lines showed differences in activity, however, they were statistically significant only in case of mint EO. Furthermore, it can be concluded that after 24 h of incubation ME vehicle itself was responsible for the observed cytotoxic effect. At the same time ME provided good solubility of constituents of EO and diminished evaporation of volatiles from culture medium. [ABSTRACT FROM AUTHOR]

A series of novel asymmetrical mono-carbonyl analogs of curcumin (AMACs) were synthesized and evaluated for cytotoxic activity using BSLT and MTT assay against Vero, HeLa, and MCF7 cell lines. The structures of the synthesized compounds were confirmed by FTIR, ¹H-NMR, 13C-NMR, and mass spectral data. The results of the cytotoxicity evaluation showed that the synthesized compounds exhibited moderate to very high toxic activity in BSLT (LC50 value 29.80-1704.23 μM); most of the compound exhibited cytotoxic activity against HeLa cell lines, which is comparable to the activity of cisplatin (IC50 value 40.65-95.55 μM), and most of the compound tested against MCF7 cell lines exhibited moderate to very high cytotoxic activity (IC50 value 7.86-35.88 μM). However, the selectivity index (SI) of the compounds was low (<1-1.96). Among the synthesized compounds, compound 1b was the most cytotoxic and selective against MCF7 cell lines. It could be considered for further development to obtain the more active and selective chemotherapeutic agents against breast cancer. [ABSTRACT FROM AUTHOR]

Oncolytic viruses gain cancer specificity in several ways. Like the majority of viruses, they grow better in cancer cells that are defective in mounting the host response to viruses. Often, they are attenuated by deletion or mutation of virulence genes that counteract the host response or are naturally occurring oncolytic mutants. In contrast, retargeted viruses are not attenuated or deleted; their cancer specificity rests on a modified, specific tropism for cancer receptors. For herpes simplex virus (HSV)-based oncolytics, the detargeting-retargeting strategies employed so far were based on genetic modifications of gD. Recently, we showed that even gH or gB can serve as retargeting tools. To enable the growth of retargeted HSVs in cells that can be used for clinical-grade virus production, a double-retargeting strategy has been developed. Here we show that several sites in the N terminus of gB are suitable to harbor the 20-amino-acid (aa)-long GCN4 peptide, which readdresses HSV tropism to Vero cells expressing the artificial GCN4 receptor and thus enables virus cultivation in the producer noncancer Vero-GCN4R cell line. The gB modifications can be combined with a minimal detargeting modification in gD, consisting in the deletion of two residues, aa 30 and 38, and replacement of aa 38 with the scFv to human epidermal growth factor receptor 2 (HER2), for retargeting to the cancer receptor. The panel of recombinants was analyzed comparatively in terms of virus growth, cell-to-cell spread, cytotoxicity, and in vivo antitumor efficacy to define the best double-retargeting strategy. [ABSTRACT FROM AUTHOR]

Insertion of a single-chain variable-fragment antibody (scFv) to HER2 (human epidermal growth factor receptor 2) in gD, gH, or gB gives rise to herpes simplex viruses (HSVs) specifically retargeted to HER2-positive cancer cells, hence to highly specific nonattenuated oncolytic agents. Clinical-grade virus production cannot rely on cancer cells. Recently, we developed a double-retargeting strategy whereby gH carries the GCN4 peptide for retargeting to the noncancer producer Vero-GCN4R cell line and gD carries the scFv to HER2 for cancer retargeting. Here, we engineered double-retargeted recombinants, which carry both the GCN4 peptide and the scFv to HER2 in gD. Novel, more-advantageous detargeting strategies were devised so as to optimize the cultivation of the doubleretargeted recombinants. Nectin1 detargeting was achieved by deletion of amino acids (aa) 35 to 39, 214 to 223, or 219 to 223 and replacement of the deleted sequences with one of the two ligands. The last two deletions were not attempted before. All recombinants exhibited the double retargeting to HER2 and to the Vero-GCN4R cells, as well as detargeting from the natural receptors HVEM and nectin1. Of note, some recombinants grew to higher yields than others. The best-performing recombinants carried a gD deletion as small as 5 amino acids and grew to titers similar to those exhibited by the singly retargeted R-LM113 and by the nonretargeted R-LM5. This study shows that double retargeting through insertion of two ligands in gD is feasible and, when combined with appropriate detargeting modifications, can result in recombinants highly effective in vitro and in vivo. [ABSTRACT FROM AUTHOR]

A new triterpenoid, 3-oxo-12β-hydroxy-oleanan-28,13β-olide (1), and six known triterpenoids 2–7 were isolated from the root bark of Ekebergia capensis, an African medicinal plant. A limonoid 8 and two glycoflavonoids 9–10 were found in its leaves. The metabolites were identified by NMR and MS analyses, and their cytotoxicity was evaluated against the mammalian African monkey kidney (vero), mouse breast cancer (4T1), human larynx carcinoma (HEp2) and human breast cancer (MDA-MB-231) cell lines. Out of the isolates, oleanonic acid (2) showed the highest cytotoxicity, i.e., IC50’s of 1.4 and 13.3 µM against the HEp2 and 4T1 cells, respectively. Motivated by the higher cytotoxicity of the crude bark extract as compared to the isolates, the interactions of oleanonic acid (2) with five triterpenoids 3–7 were evaluated on vero cells. In an antiplasmodial assay, seven of the metabolites were observed to possess moderate activity against the D6 and W2 strains of P. falciparum (IC50 27.1–97.1 µM), however with a low selectivity index (IC50(vero)/IC50(P. falciparum-D6) < 10). The observed moderate antiplasmodial activity may be due to general cytotoxicity of the isolated triterpenoids.

نئوسپورا کنینوم (Neospora caninum) یک تک یاخته داخل سلولی اجباری و عامل بیماریزای مهم در گاو و سگ بشمار می رود. امروزه به طور گسترده ای از کشت سلولی به منظور جداسازی و تکثیر نئوسپورا کنینوم برای اهداف مختلفی استفاده می شود. در اکثر مطالعات، سلول Vero (با منشا سلول های کلیه میمون سبز آفریقایی) به عنوان بهترین سلول برای جداسازی و تکثیر نئوسپورا معرفی شده است. در این مطالعه حساسیت سلول های مختلف به این انگل و توان تکثیر آن در رده های مختلف سلولی مورد ارزیابی قرار گرفته است. بدین منظور در ابتدا تاکی زوئیت های انگل به منظور آداپته سازی به رده های سلولی SW742,Vero ,TLI ,MA-104, MDCk ,McCoy افزوده شد و به مدت دو هفته کشت داده شد. سپس میزان یکسانی از تاکی زوئیت در مجاورت میزان یکسانی از هر کدام از رده های سلولی نامبرده قرار گرفت و تا زمان ایجاد CPE کامل در همه سلول ها، روزانه (به مدت 5 روز) 10 فیلد از فلاسک کشت سلولی در زیر میکروسکوپ اینورت مشاهده و تاکی زوئیت های رها شده در آن، شمارش شدند. در روز چهارم بیشترین میزان تاکی زوئیت های رها شده از سلول ها مشاهده شد و در روز پنج رو به کاهش رفت. میانگین تاکی زوئیت های رها شده از سلول ها در روز چهارم بعد از انکوباسیون، حساسیت خوبی را در سلول (24.2×104)Vero، (22.1×104)MA-104، (21.2×104) SW742 و (20.58×104) TLI نشان می دهد. در حالیکه سلول های (MDCK 8125) و (McCoy 9375) حساسیت و توان تکثیر کمی برای نئوسپورا از خود نشان دادند. با توجه با این نتایج سلول MA-104 و TLI و SW742 سلول های مناسبی جهت جداسازی و تکثیر نئوسپورا می باشد. در ضمن با توجه به این که سلول TLI (لنفوسیت ترانسفورم شده با تیلریا) یک سلول معلق می باشد کار کردن با این سلول به مراتب راحت تر، سریعتر و کم هزینه تر از سلول های چسبنده می باشد.