Secondary metabolite profile of Streptomyces spp. changes when grown with the sub-lethal concentration of silver nanoparticles: possible implication in novel compound discovery.

The decline of new antibiotics and the emergence of multidrug resistance in pathogens necessitates a revisit of strategies used for lead compound discovery. This study proposes to induce the production of bioactive compounds with sub-lethal concentrations of silver nanoparticles (Ag-NPs). A total of Forty-two Actinobacteria isolates from four Saudi soil samples were grown with and without sub-lethal concentration of Ag-NPs (50 µg ml-1). The spent broth grown with Ag-NPs, or without Ag-NPs were screened for antimicrobial activity against four bacteria. Interestingly, out of 42 strains, broths of three strains grown with sub-lethal concentration of Ag-NPs exhibit antimicrobial activity against Staphylococcus aureus and Micrococcus luteus. Among these, two strains S4-4 and S4-21 identified as Streptomyces labedae and Streptomyces tirandamycinicus based on 16S rRNA gene sequence were selected for detailed study. The change in the secondary metabolites profile in the presence of Ag-NPs was evaluated using GC-MS and LC-MS analyses. Butanol extracts of spent broth grown with Ag-NPs exhibit strong antimicrobial activity against M. luteus and S. aureus. While the extracts of the controls with the same concentration of Ag-NPs do not show any activity. GC-analysis revealed a clear change in the secondary metabolite profile when grown with Ag-NPs. Similarly, the LC-MS patterns also differ significantly. Results of this study, strongly suggest that sub-lethal concentrations of Ag-NPs influence the production of secondary metabolites by Streptomyces. Besides, LC-MS results identified possible secondary metabolites, associated with oxidative stress and antimicrobial activities. This strategy can be used to possibly induce cryptic biosynthetic gene clusters for the discovery of new lead compounds.

In Vivo Study of Moringa oleifera Seed Extracts as Potential Sources of Neuroprotection against Rotenone-Induced Neurotoxicity.

Parkinson's disease (PD) is a leading neurodegenerative disorder affecting 1-3 percent of the elderly population. Oxidative stress is the primary factor for the neurodegeneration of Substantia Nigra (SN). The current study aims to assess the seed extracts of Moringa oleifera (MO) on rotenone-mediated motor function impairments in a PD mouse model. For this purpose, two different seed extracts of MO were prepared, including aqueous MO (AqMO) and ethanolic MO (EthMO). Male Swiss albino mice were grouped into five groups. Mice received 2.5 mg/kg rotenone for 21 consecutive days, and control mice received the vehicle. Extract-treated mice received 200 mg/kg AqMO and EthMO separately, orally and daily for 28 days. Sinemet-treated mice received 20 mg/kg, oral dose, as a positive group. The motor function performance was evaluated using standard neurobehavioral tests. The antioxidant potentials of MO seed extracts were estimated by lipid peroxidation (LPO), reduced glutathione (GSH), glutathione-s-transferase (GST) and catalase (CAT) activities in mice brain homogenates. The PD mice brain SN sections were investigated for neurodegeneration. MO seed extract-treated mice showed a significant reduction in motor dysfunction compared to rotenone-treated mice as assessed through the open field, beam walk, pole climb-down, tail suspension, stride length and stepping tests. Increased antioxidant capacities of the PD mice brains of MO extract-administered groups were observed compared to the control. A histological study showed reduced signs of neurodegeneration, vacuolation around multipolar cells and cytoplasmic shrinkage in MO extract-treated mice SN brain sections. Collectively, MO seed extracts protected the animals from locomotor deficits induced by rotenone, possibly through antioxidant means, and seem to have potential applications in neurodegenerative diseases.

Chemical Characterization and Chemotaxonomic Significance of Essential Oil Constituents of Matricaria aurea Grown in Two Different Agro-Climatic Conditions.

A comprehensive study on chemical characterization of essential oil (EO) constituents of a rarely explored plant species (Matricaria aurea) of the Asteraceae family grown in Saudi Arabia and Jordan was carried out. Analyses were conducted employing gas chromatographic approaches such as GC-MS, GC-FID, and Co-GC, as well as RT, LRI determination, and database and literature comparisons, on two diverse stationary phase columns, which led to the identification of a total of 135 constituents from both EOs. Oxygenated sesquiterpenes were found to be the most predominant chemical class of Saudi M. aurea EOs, in which α-bisabolol (27.8%), γ-gurjunenepoxide (21.7%), (E, E)-α-farnesene (16.3%), and cis-spiroether (7.5%) were present as major components. In contrast, the most dominant chemical class of Jordanian M. aurea oil was found to be sesquiterpene hydrocarbons, where (E, E)-α-farnesene (50.2%), γ-gurjunenepoxide (8.5%), (E)-ß-farnesene (8.1%), and (Z, E)-α-farnesene (4.4%) were detected as chief constituents. It is interesting to mention here that Saudi and Jordanian M. aurea EOs showed quite interesting chemical compositions and were found to have different chemotypes when compared to previously reported M. aurea EO compositions.

CBPDdb: a curated database of compounds derived from Coumarin-Benzothiazole-Pyrazole.

The present article describes the building of a small-molecule web server, CBPDdb, employing R-shiny. For the generation of the web server, three compounds were chosen, namely coumarin, benzothiazole and pyrazole, and their derivatives were curated from the literature. The two-dimensional (2D) structures were drawn using ChemDraw, and the .sdf file was created employing Discovery Studio Visualizer v2017. These compounds were read on the R-shiny app using ChemmineR, and the dataframe consisting of a total of 1146 compounds was generated and manipulated employing the dplyr package. The web server is provided with JSME 2D sketcher. The descriptors of the compounds are obtained using propOB with a filter. The users can download the filtered data in the .csv and .sdf formats, and the entire dataset of a compound can be downloaded in .sdf format. This web server facilitates the researchers to screen plausible inhibitors for different diseases. Additionally, the method used in building the web server can be adapted for developing other small-molecule databases (web servers) in RStudio. Database URL: https://srampogu.shinyapps.io/CBPDdb_Revised/.

Evaluation of Photocatalytic, Antioxidant, and Antibacterial Efficacy of Almond Oil Capped Zinc Oxide Nanoparticles.

In this study, ZnO nanoparticles (NPs) were synthesized in the presence of almond oil at various molar ratios of zinc acetate and sodium hydroxide, including 0.5:1, 0.75:1, 1:1, 1.25:1, and 1.5:1, to obtain pH values of 11, 10, 9, 8, and 7, respectively. The XRD results revealed that ZnO NPs exhibit a hexagonal structure, with high crystallinity. SEM results showed that dense and large sized ZnO NPs were formed at pH 11, and relatively small (~30-40 nm) NPs were obtained at pH 9. The size distribution can be explained in terms of the presence of OH- ions at different pH levels. However, the larger size of the NPs at pH 7 compared to those at pH 8-11 were due to the coalescence of NPs suitable for antioxidant/antibacterial activities. ZnO NPs demonstrated a high degradation efficiency (~93%) in 90 min, with a high rate constant for Methyl Orange (MO), which is better than the previously reported rate. The larger sized almond oil capped ZnO NPs also showed excellent radical scavenging activity (94%) and are proven to be good carriers to resist Escherichia coli (E. coli) bacteria.

Cerium Oxide/Graphene Oxide Hybrid: Synthesis, Characterization, and Evaluation of Anticancer Activity in a Breast Cancer Cell Line (MCF-7).

In the present study, we used a simple ultrasonic approach to develop a Cerium oxide/Graphene oxide hybrid (CeO2/GO hybrid) nanocomposite system. Particle size analysis, Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and X-ray Diffraction (XRD) have been used to analyze the physio-chemical characteristics of the developed nanocomposite. The synthesized hybrid system has also been examined to assess its anticancer capability against MCF-7 cell lines and normal cell lines at different sample concentrations, pH values, and incubation intervals using an antiproliferative assay test. The test results demonstrate that as sample concentration rises, the apoptotic behavior of the CeO2/GO hybrid in the MCF-7 cell line also rises. The IC50 was 62.5 µg/mL after 72 h of incubation. Cytotoxicity of cisplatin bound CeO2/GO hybrid was also tested in MCF-7 cell lines. To identify apoptosis-associated alterations of cell membranes during the process of apoptosis, a dual acridine orange/ethidium bromide (AO/EB) fluorescence staining was carried out at three specified doses (i.e., 1000 µg/mL, 250 µg/mL, and 62.5 µg/mL of CeO2/GO hybrid). The color variations from both live (green) and dead (red) cells were examined using fluorescence microscopy under in vitro conditions. The quantitative analysis was performed using flow cytometry to identify the cell cycle at which the maximum number of MCF-7 cells had been destroyed as a result of interaction with the developed CeO2/GO hybrid (FACS study). According to the results of the FACS investigation, the majority of cancer cells were inhibited at the R3 (G2/M) phase. Therefore, the CeO2/GO hybrid has successfully showed enhanced anticancer efficacy against the MCF-7 cell line at the IC50 concentration. According to the current study, the CeO2/GO platform can be used as a therapeutic platform for breast cancer. The synergetic effects of the developed CeO2/GO hybrid with the MCF-7 cell line are presented.

Diversity of Citrullus colocynthis (L.) Schrad Seeds Extracts: Detailed Chemical Profiling and Evaluation of Their Medicinal Properties.

Seeds and fruits of Citrullus colocynthis have been reported to possess huge potential for the development of phytopharmaceuticals with a wide range of biological activities. Thus, in the current study, we are reporting the potential antimicrobial and anticancer properties of C. colocynthis seeds extracted with solvents of different polarities, including methanol (M.E.), hexane (H.E.), and chloroform (C.E.). Antimicrobial properties of C. colocynthis seeds extracts were evaluated on Gram-positive and Gram-negative bacteria, whereas, anticancer properties were tested on four different cell lines, including HepG2, DU145, Hela, and A549. All the extracts have demonstrated noteworthy antimicrobial activities with a minimum inhibitory concentration (MIC) ranging from 0.9-62.5 µg/mL against Klebsiella planticola and Staphylococcus aureus; meanwhile, they were found to be moderately active (MIC 62.5-250 µg/mL) against Escherichia coli and Micrococcus luteus strains. Hexane extracts have demonstrated the highest antimicrobial activity against K. planticola with an MIC value of 0.9 µg/mL, equivalent to that of the standard drug ciprofloxacin used as positive control in this study. For anticancer activity, all the extracts of C. colocynthis seeds were found to be active against all the tested cell lines (IC50 48.49-197.96 µg/mL) except for the chloroform extracts, which were found to be inactive against the HepG2 cell line. The hexane extract was found to possess the most prominent anticancer activity when compared to other extracts and has demonstrated the highest anticancer activity against the DU145 cell line with an IC50 value of 48.49 µg/mL. Furthermore, a detailed phytoconstituents analysis of all the extracts of C. colocynthis seeds were performed using GC-MS and GC-FID techniques. Altogether, 43 phytoconstituents were identified from the extracts of C. colocynthis seeds, among which 21, 12, and 16 components were identified from the H.E., C.E., and M.E. extracts, respectively. Monoterpenes (40.4%) and oxygenated monoterpenes (41.1%) were the most dominating chemical class of compounds from the hexane and chloroform extracts, respectively; whereas, in the methanolic extract, oxygenated aliphatic hydrocarbons (77.2%) were found to be the most dominating chemical class of compounds. To the best of our knowledge, all the phytoconstituents identified in this study are being reported for the first time from the C. colocynthis.

Green Synthesized Silver Nanoparticle-Loaded Liposome-Based Nanoarchitectonics for Cancer Management: In Vitro Drug Release Analysis.

Silver nanoparticles act as antitumor agents because of their antiproliferative and apoptosis-inducing properties. The present study aims to develop silver nanoparticle-loaded liposomes for the effective management of cancer. Silver nanoparticle-encapsulated liposomes were prepared using the thin-film hydration method coupled with sonication. The prepared liposomes were characterized by DLS (Dynamic Light Scattering analysis), FESEM (Field Emission Scanning Electron Microscope), and FTIR (Fourier Transform Infrared spectroscopy). The in vitro drug release profile of the silver nanoparticle-loaded liposomes was carried out using the dialysis bag method and the drug release profile was validated using various mathematical models. A high encapsulation efficiency of silver nanoparticle-loaded liposome was observed (82.25%). A particle size and polydispersity index of 172.1 nm and 0.381, respectively, and the zeta potential of -21.5 mV were recorded. FESEM analysis revealed spherical-shaped nanoparticles in the size range of 80-97 nm. The in vitro drug release profile of the silver nanoparticle-loaded liposomes was carried out using the dialysis bag method in three different pHs: pH 5.5, pH 6.8, and pH 7.4. A high silver nanoparticle release was observed in pH 5.5 which corresponds to the mature endosomes of tumor cells; 73.32 ± 0.68% nanoparticle was released at 72 h in pH 5.5. Among the various mathematical models analyzed, the Higuchi model was the best-fitted model as there is the highest value of the correlation coefficient which confirms that the drug release follows the diffusion-controlled process. From the Korsmeyer-Peppas model, it was confirmed that the drug release is based on anomalous non-Fickian diffusion. The results indicate that the silver nanoparticle-loaded liposomes can be used as an efficient drug delivery carrier to target cancer cells of various types.

Development and Validation for Quantification of 7-Nitroso Impurity in Sitagliptin by Ultraperformance Liquid Chromatography with Triple Quadrupole Mass Spectrometry.

The purpose of this research study was to develop an analytical method for the quantification of 7-nitroso-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4] triazolo [4,3-a] pyrazine (7-nitroso impurity), which is a potential genotoxic impurity. Since sitagliptin is an anti-diabetic medication used to treat type 2 diabetes and the duration of the treatment is long-term, the content of nitroso impurity must be controlled by using suitable techniques. To quantify this impurity, a highly sensitive and reproducible ultraperformance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed. The analysis was performed on a Kromasil-100, with a C18 column (100 mm × 4.6 mm with a particle size of 3.5 µm) at an oven temperature of approximately 40 °C. The mobile phase was composed of 0.12% formic acid in water, with methanol as mobile phases A and B, and the flow rate was set to 0.6 mL/min. The method was validated according to the current International Council for Harmonisation (ICH) guidelines with respect to acceptable limits, specificity, reproducibility, accuracy, linearity, precision, ruggedness and robustness. This method is useful for the detection of the impurity at the lowest limit of detection (LOD), which was 0.002 ppm, and the lowest limit of quantification (LOQ), which was 0.005 ppm. This method was linear in the range of 0.005 to 0.06 ppm and the square of the correlation coefficient (R2) was determined to be > 0.99. This method could help to determine the impurity in the regular analysis of sitagliptin drug substances and drug products.

Comprehensive Phytochemical Analysis of Various Solvent Extracts of Artemisia judaica and Their Potential Anticancer and Antimicrobial Activities.

Solvents play an important role in the extraction process by considerably affecting the amount and nature of secondary metabolites of medicinal plants. Thus, the effect of solvents must be investigated to obtain desired biological properties of plant extracts. In the current study, we extracted aerial parts of Artemisia judaica, native to Saudi Arabia, in three different solvents, including methanol (MeOH), hexane (Hex), and chloroform (Chl). Obtained extracts from the aerial parts of A. judaica were analysed by GC-MS and GC-FID techniques, which resulted in the identification of 46, 18, and 17 phytoconstituents from the Hex, Chl, and MeOH extracts, respectively. All the extracts contain oxygenated terpenes, aliphatic hydrocarbons, and aromatics as major classes of compounds in varying amounts. Among the various phytoconstituents identified, piperitone was the dominant compound and was found in all the extracts in different amounts, specifically, 28.8, 26.1, and 20.1% in the Chl, MeOH, and Hex extracts, respectively. Moreover, all these extracts (Chl, MeOH, and Hex) were tested for the antimicrobial properties on both Gram-positive and negative bacteria as well as for their anticancer properties on four different cell lines including HepG2, DU145, Hela, and A549. Among the different extracts, the Hex and Chl extracts demonstrated identical antimicrobial properties, while the Chl extract showed superior anticancer properties when compare to the other extracts. The higher biological properties of Chl extracts including both antimicrobial and anticancer activities may be attributed to the presence of large amounts of piperitone and/or santonin, which are distinctly present in excess amounts in the Chl extract.