Current Pharmaceutical Biotechnology - Volume 22, Issue 10, 2021

Background: Oleanolic acid is an oleanane triterpene found in many plant species all over the world. This compound is also a major saponin in leaves of Polyscias fruticosa and possesses several promising pharmacological activities, such as hepatoprotective effects, and antiinflammatory, antioxidant, or anticancer activities. Objective: The objective of the present work is to establish cell suspension culture of P. fruticosa, investigate the influence of several factors such as plant growth regulators and carbon source on cell growth, and determine their oleanolic acid content. Methods: Cell culture was established by using 2 g fresh weight of 30 day old friable callus derived from in vitro stem segment in 50 mL of liquid medium with a shaking speed of 220 rpm. The culture was then incubated at 25±2ºC with a shaking speed of 120 rpm in the period of 12 h daylight at a light intensity of about 6.75 μmol/m2/s. Cell growth was measured by fresh and dry biomass at 16 h day. Oleanolic acid content was determined using HPLC analysis. Results & Discussion: The study results showed that MS medium containing 2% sucrose as a carbon source, supplemented with 1 mg/L 6-benzylaminopurine and 0.5 mg/L 2,4-dichlorophenoxyacetic acid was the most appropriate growth medium. Cell biomass and oleanolic acid content reached the highest values of 0.43 g dry weight/flask and 25.4 mg/g dry weight, respectively. Conclusion: These results indicated the potential production of oleanolic acid, a compound with high pharmacological value, from P. fruticosa cell culture.

Background: Aptamers are short single-stranded oligonucleotides; due to their 3D structure, they can specifically bind to various targets. They can target several molecules ranging from metal ions, organic components to proteins, and large cells. Methods: According to the high affinity of aptamers, they can be used for diagnosis, therapeutic, vaccine development, and gene silencing applications. The conventional method for aptamer selection is known as the Systematic Evolution of Ligands by Exponential (SELEX). However, despite the efficiency of SELEX as a screening procedure, it is beneficial to develop more rational procedures for aptamer selection. Results & Discussion: Herein, in silico approaches can play an effective role given their potential in representing an efficient, cost-effective, parallelizable, and rapid strategy. In recent years, several attempts have been applied to develop algorithms and software for the rational selection of aptamers. However, there is still a need for more efforts to achieve the most efficient techniques in this area. Conclusion: In this review, we aim to overview different computational approaches that are used for aptamer selection.

Background: Colon cancer is the second leading cause of cancer related deaths worldwide with about 1.2 million new cases identified annually. While considering swift progress in the field of molecular biology, new horizons in the treatment approaches have been materialized in colon cancer with conventional methods being replaced with targeted therapies. Methods: In this review, we focused on the existing conventional therapies utilized for colon cancer by comparing the effectiveness of various standard/conventional therapies with respect to overall survival parameter. Regardless of all the conventional treatments and scientific research, the disease remains to be the one of the major cause of cancer related death and rising as societal burden due to its co morbidities. Thus, we have also discussed briefly in this review, all the possible biotechnological next-generation therapeutics including nucleic acid medicines, CRISPR-Cas9 technology, adoptive cell therapy, cancer stem cells and therapy, gut microbiome, and personalized medicines, which might be promising after effective clinical trials. Results: From our study, we suggest that the use of neoadjuvant chemoradiotherapy in resected patients was found to be safe and effective therapy in treating colon cancer and thereby improving overall survival in patients. From considering the total estimate of our meta-analysis plot, we state that the existing therapies are not much satisfying to improve the overall survival and more research has to be carried out in this field to find an effective therapy to treat colon cancer. Conclusions: As existing therapies are not much satisfying and are unable to improve the overall survival, we brought together a diversity of possible approaches focusing on biotechnological nextgeneration therapeutics to treat colon cancer. Hence, various multi-disciplinary choices are mandatory in order to provide patients with distended access to tailored treatments.

Ischemic stroke is one of the major causes of deaths around the world. Prominent changes, including emotional, cognitive, and behavioral, in the Central Nervous System (CNS) are observable during the recovery phase of a stroke. Brain ischemia affects a large portion of patients across the world with permanent disability or death. During the ischemic stroke onset, apart from inducing brain damage, existing microglial cells, immune cells, inflammatory factors such as chemokines and cytokines affect the process of brain repair. For the reduction of this damage, there is a need to understand the basic mechanisms involved in ischemic stroke and how to repair it with the help of efficient therapeutic strategies. At present, anti-inflammatory therapeutic measures are the most potent alternatives for its prevention. Due to the existence of various drawbacks in this therapy, newer remedial measures are required to overcome this situation. The most economically efficient treatment option, with lesser side effects, is the utilization of medicinal plants. Worldwide, plants are utilized for the treatment of various diseases such as bacterial infections, skin disorders, cancer, asthma, respiratory problems, etc. Various medicinal plants have been administered over a long period, and they have been proven to cure ischemic strokes. Therefore, in this review, an overview of various plants and their phytocompounds, which are responsible for the treatment of ischemic stroke, has been discussed.

Background: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the causative agent of the current pandemic of Coronavirus Disease-2019 (COVID-19). The progression of COVID-19 is related to an excessive host inflammatory immune response to SARS-CoV-2 infection, which is considered a major cause of disease severity and death. Dysregulated immune response produces huge amounts of pro-inflammatory cytokines and chemokines called "cytokine storm". Moreover, the activation of microthrombi formation plays an important role in multiple organ failure. Methods: Keeping into consideration the potent anti-inflammatory activity of black seed and its major constituent Thymoquinone (TQ), we hypothesize their potential implication in the treatment of COVID-19 patients. A literature search was performed in PubMed, ScienceDirect, Google Scholar and Scopus electronic databases using the terms, including black seed, N. sativa, thymoquinone, SARSCoV- 2, COVID-19 and inflammatory immune response. Results: Various studies confirmed that Black seed and TQ reduced the thrombus formation, the expression of tissue factor and the immune activation. Furthermore, TQ demonstrated the broad-spectrum antimicrobial activity that may be effective in controlling the secondary infections in COVID-19 patients. Conclusion: Keeping into consideration the multi-targeting nature of the black seed and TQ, they may be used as a potential therapeutic formulation or as an adjunct therapy in the treatment of COVID-19 patients.

Background: Nitrate and nitrite can get into the body through the consumption of contaminated water either directly or indirectly. The accumulation of these compounds in the body, in the long run, leads to health problems, for example, digestive disorders, cancers, and even death threats in children. The aim of this review was to investigate nitrate and nitrite pollution levels in drinking water and fruit juices in Iran. Methods: In this review, data were collected through searching the Scientific Information Database, Science-Direct, Scopus, PubMed, Google Scholar, and Magiran databases using the keywords Nitrate, Nitrite, Drinking water, Drinking Water Resources, Juice and Iran. Finally, the location of the studies was geocoded through the Google My Maps (https://www.google.com/mymaps) software. Results: Studies clearly indicated that the juices are safe in terms of nitrate. Nitrate and nitrite values were less than the national and international standards in all samples of bottled drinking water except for a few of the studies. The results of the reviewed studies also indicated that the nitrate content was higher than that written on the label in 96% of the samples, and nitrite was not labeled in 80% of them. The nitrate quantity was higher than the permissible limit in the water distribution network of Bushehr, Gilan and Mazandaran Provinces. Talesh, Ardabil, Hashtgerd, Divandareh, and Kerman cities had high nitrate levels in more than 50% of wells. Conclusion: Using nitrogen fertilizers and the lack of a wastewater treatment system were the main reasons for the presence of nitrate and nitrite.

Objective: To summarize and assess the effects of probiotic preparations on inflammatory cytokine levels in patients with Chronic Kidney Disease (CKD). Methods: We searched through the PubMed, Cochrane Library, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature Database (CBM), and Wan Fang databases for Randomized Controlled Trials (RCTs) that report the impact of probiotic preparations on inflammatory cytokines in CKD patients. Outcomes were composed of serum levels of CReactive Protein (CRP), Interleukin 6 (IL-6), Tumor Necrosis Factor-α (TNF-α), serum urea, creatinine, uric acid, Para-Cresol Sulfate (PCS), and Indoxyl-Sulfate (IS). The Mean Differences (MDs) with 95% Confidence Intervals (CIs) were considered as effect estimates. Sensitivity analysis and Egger’s linear regression test were performed to evaluate the stability of results and publication bias. This study was registered with PROSPERO number CRD42020176557. Results and Discussion: Sixteen studies met the inclusion criteria. Evidence showed that serum CRP levels were decreased in the intervention group (WMD, -12.29, 95% CI, -16.41 to -8.16, p = 0). The IL-6 was significantly reduced only in the prebiotic group (SMD, -0.73, 95% CI, -1.3 to -0.16, p = 0.012). However, no reduction was observed in TNF-α (SMD, -0.07, 95% CI, -0.51 to 0.38, p = 0.772). Moreover, there was no significant change in serum uremic toxin, including creatine, urea, uric acid, PCS, and IS. Conclusion: Probiotic preparations decrease the serum levels of inflammatory cytokines in CKD patients but do not affect the serum uremic toxin levels. The results of this meta-analysis suggest essential guidance for treatment decisions in clinical practice.

Background: COVID-19 caused by SARS-CoV-2 has been declared as a global pandemic by WHO. Comprehensive analysis of this unprecedented outbreak may help to fight against the disease and may play a pivotal role in decreasing the mortality rate linked with it. Papain-like protease (PLpro), a multifunctional polyprotein, facilitates the replication of SARS-CoV-2 and evades it from the host immunological response by antagonizing cytokines, interferons and may be considered as a potential drug target to combat the current pandemic. Methods: Natural moieties obtained from medicinal plants were analysed for their potency to target PLpro of SARS-CoV-2 by molecular docking study and were compared with synthetic analogs named as remdesivir, chloroquine and favipiravir. The stability of complexes of top hits was analysed by MD Simulation, and interaction energy was calculated. Furthermore, average RMSD values were computed and deepsite ligand-binding pockets were predicted using Play Molecule. Drug-like-abilities of these moieties were determined using ADMET and bond distance between the ligand and active site was assessed to predict the strength of the interaction. Results: Nimbocinol (-7.6 Kcal/mol) and sage (-7.3 Kcal/mol) exhibited maximum BA against PLpro SARS-CoV-2 as evident from molecular docking study, which was found to be even better than remdesivir (-6.1 Kcal/mol), chloroquine (-5.3 Kcal/mol) and favipiravir (-5.7 Kcal/mol). Both nimbocinol- PLpro and sage-PLpro SARS-CoV-2 complex exhibited stable conformation during MD Simulation of 101ns at 310 K, and potential, kinetic and electrostatic interaction energies were computed, which was observed to be concordant with results of molecular docking study. RMSD average values were found to be 0.496 ± 0.015 Å and 0.598 ± 0.023 Å for nimbocinol and sage, respectively, thus revealing that both the deviation and fluctuations during MD Simulation were observed to be least. Deepsite prediction disclosed that both compounds occupied cryptic pockets in receptor and non-bond distance analysis revealed the formation of hydrogen bonds during ligand-receptor interaction. ADMET exploration further validated the drug-like properties of these compounds. Conclusion: Present study revealed that active constituents of Azadirachta indica and Salvia officinalis can be potentially used to target SARS-CoV-2 by hindering its replication process.

Background: The aim of this research is to investigate the potential activity of Salvia officinalis and various hair samples responsible for secretion of essential oil. In Tunisia, biological activity of Salvia officinalis is poorly recorded. Salvia leaves contain various types of hairs (glandular and nonglandular). Methods: The investigation of different trichomes was carried out by Scanning Electron Microscope (SEM) apparatus. Antiradical potential was assessed using 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assay. Antimicrobial activity was analysed using disc diffusion assay. The extracts of Salvia officinalis (SvOf), showed the following order of richness in phenolic contents: methanolic (70.76 mg GAE/g DW), aqueous (43.76 mg GAE/g DW) and infusion (9.42 mg GAE/g DW). The methanolic fraction records the highest levels of flavonoids (77 mg QE/g DW) compared with the aqueous extracts (33.19 mg EQ/g DW) and infusion (26.25 mg EQ/g DW). MESvOf showed higher value of free radical scavenging activity towards DPPH free radical and ferric reducing power. Results: The results revealed that the main bioactive constituents in the methanolic fractions of sage leaves generally exhibited higher antibacterial effects. Overall, sage phytocompounds constitute a promising approach for the treatment of infectious diseases. Discussion: Functional groups detected in S. officinalis by Fourier Transform Infrared Spectrophotometer (FT-IR) were mainly phenols, saccharides, amine and Aromatic (Ar-H). Conclusion: Antiradical and antibacterial activities of Salvia officinalis are mainly due to phenolic content and other bioactive compounds. Non-glandular hairs are the most important trichomes in the Salvia leaves.

Introduction: Microglia-mediated inflammatory responses play a crucial role in aging-related neurodegenerative diseases. The TXNIP/NLRP3 pathway is a key pathway leading to microglial activation. Panax notoginseng Saponins (PNS) have been widely used for the treatment of stroke in China. Objective: This study evaluates the anti-neuroinflammatory effect of PNS and investigates the mechanism via TXNIP-mediated NLRP3 inflammasome activation in aging rats. Material and Methods: Eighteen-month-old Sprague-Dawley rats were randomly divided into the aging control group and PNS treated groups (n=15 each group). For PNS-treated groups, rats were administrated food with PNS at the doses of 10 mg/kg and 30 mg/kg for consecutive 6 months until they were 24-month old. Rats from the aging control group were given the same food without PNS. Twomonth- old rats were purchased and given the same food until they were 6-months old as the adult control group (n = 15). Then, the cortex and hippocampus were rapidly harvested and deposited. H staining was used to assess histo-morphological changes. Western blotting was carried out to detect the protein expression. Immunofluorescence was employed to measure the co-localization of NLRP3, TXNIP and Iba-1. In vitro model was established by LPS+ATP co-incubation in the BV2 microglia cell line. Results: Aging rats exhibited increased activation of microglia, accompanied by a high level of IL-1β expression. Meanwhile, aging rats showed enhanced protein expression of TXNIP and NLRP3 related molecules, which co-localized with microglia. PNS treatment effectively reduced the number of degenerated neurons and reversed the activation of the TXNIP/NLRP3 inflammatory pathway. In vitro results showed that PNS up to 100 μg/ml had no significant toxicity on BV2 microglia. PNS (25, 50 μg/ml) effectively reduced the inflammatory response induced by LPS and ATP co-stimulation, thus inhibiting the expression of TXNIP/NLRP3 pathway-related proteins. Discussion and Conclusion: PNS treatment improved aging-related neuronal damage through inhibiting TXNIP mediated NLRP3 inflammasome activation, which provided a potential target for the treatment of inflammation-related neurodegenerative diseases.

Background: Cistanche tubulosa is a tonic in traditional Chinese medicines and has a broad spectrum of biological activity, including anti-inflammatory. However, the anti-inflammatory major constituents of C. tubulosa and their underlying mechanisms are still unknown. Objective: The aim of the current study was to explore the separation and structural characterization of lignan glycosides from C. tubulosa (Schenk) Wight., their anti-inflammatory activity and the underlying mechanism. Materials and Methods: Fractionation and isolation of the 85% EtOH extract of C. tubulosa (Schenk) Wight. were carried out and the primary ingredients lignan glycosides (1-6) were structurally characterized. CCK8 methods were used to evaluate the cytotoxic effect of lignan glycosides (1-6). Effects of lignan glycosides (1-6) on NO production in LPS/IFN-γ-induced RAW264.7 macrophages cells were measured using Griess reagent by reaction with nitrite. The mRNA expression levels of iNOS, COX-2, IL-1β, IL-6, TNF-a, and TGF-β treated RAW264.7 cells with various concentrations (0, 25 and 50 μg/ml) of lignan glycosides (1, 4) in the presence of LPS (10 ng/ml) and IFN-γ (20 ng/ml) for 24 h were analyzed by quantitative RT-PCR. Also, the protein expressions of iNOS, COX-2, PI3K, AKT, p-AKT and β -actin were determined using Western blot analysis. A molecular docking study was performed to investigate the interactions between the lignan glycosides and the PI3K using Autodock vina 1.1.2 package. Results: Six lignan glycosides (1-6) were isolated from stems of C. tubulosa. Among them, (+)- pinoresinol-4-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (5) and eleutheroside E (6) were firstly isolated from C. tubulosa. Of these lignans, 1 and 4 exhibited pronounced inhibitions on NO production with the values of 33.63 ± 4.78 and 39.28 ± 5.52 % at 50 μg/ml, respectively. Additionally, LPS/IFN-γ-induced expression of inducible Nitric Oxide Synthase (iNOS), Cyclooxygenase-2 (COX-2), Interleukin-1β (IL-1β), IL-6, and Tumor Necrosis Factor-a (TNF-a) was significantly suppressed by pre-treatment of 1 and 4 in a dose-dependent manner. While 1 and 4 increased the mRNA levels of anti-inflammatory cytokines (TGF-β). Furthermore, 1 and 4 significantly inhibited the protein levels of PI3K and p-AKT in a dose-dependent manner. Conclusion: Taken together, these results suggest that 1 and 4 play an important role in the attenuation of LPS/IFN-γ-induced inflammatory responses in RAW264.7 cells and that the mechanisms involve down-regulation of the PI3K/AKT pathway.