Pre-clinical studies comparing the anti-inflammatory potential of artemisinic compounds by targeting NFκB/TNF-α/NLRP3 and Nrf2/TRX pathways in Balb/C mice.

Artemisinin, artemether, artesunate, and dihydroartemisinin are renowned for their antimalarial potential. The current study aims to repurpose the above-mentioned artemisinic compounds (ACs) by conducting an intercomparison to evaluate their antiinflammatory potential (AIP). In order to develop potential candidates for the evaluation of AIP of ACs (50 and 100 mg/kg BW), carbon tetrachloride (1ml/kg body weight (BW)) was administered intraperitoneally to BALB/c mice. Alterations in animal behavior were assessed weekly through tail suspension test, force swim test, open field test, Y-maze test, inverted screen analysis, and weight lifting test. Aberrations in hematological, serological, endogenous antioxidants, and oxidative stress marker profiles were assessed in all twelve groups. Histological alterations were read using hematoxylin and eosin staining. Levels of inflammatory markers including nuclear factor kappa B (NF-κB), tumor necrosis factor alpha (TNF-α), and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), were determined using immunohistochemical analysis (IHCA). Antioxidant markers i.e., nuclear factor erythroid-2-related factor (Nrf-2) and thioredoxin (TRX) were also quantified through IHCA. Comet assay was performed to quantify DNA damage. Oral administration of ACs to mice significantly alleviated the carbon tetrachloride induced inflammation in comparison with silymarin. Reduced levels of several inflammatory markers including nitric oxide, thiobarbituric acid reactive substances, interleukin-1 beta, NF-κB, TNF-α, and NLRP3, underscore the substantial AIP of ACs. IHCA depicted the revitalized percent relative expression of Nrf-2 and TRX in groups treated with ACs. Behavioral analysis revealed that ACs-treated groups significantly (p<0.05) attenuated the memory deficit, anxiety, and depressive-like behavior. Moreover, histopathological, hematological, serological, and endogenous antioxidant profiles indicated substantial AIP of ACs. Findings of comet assay further bolstered the compelling evidence as DNA damage was significantly (p<0.05) curbed down after ACs (100 mg/kg) treatment. All these outcomes implied that ACs exhibited AIP in a dose-dependent manner with maximal AIP imparted by artemisinin (100 mg/kg). This pre-clinical investigation avers the tremendous AIP of ACs targeting key molecular pathways. The current study divulges artemisinin as the most potent antiinflammatory agent among the tested compounds.

Mechanistic insight into the synergistic antimicrobial potential of Fagonia indica Burm.f. extracts with cefixime.

Fagonia indica Burm.f. is known for its anti-infective character and has been studied in the present work as a synergistic remedy against resistant bacterial strains. Initially, phytochemicals were quantified in n-Hexane (n-Hex), ethyl acetate (E.A), methanol (MeOH), and aqueous (Aq.) extracts by Total Phenolic Content (TPC), Total Flavonoid Content (TFC) and Reverse Phase High Performance Liquid Chromatography (RP-HPLC) analysis. Later, after establishing an antibacterial resistance profile for extracts and antibiotics against gram-positive and gram-negative strains, synergism was evaluated in combination with cefixime through time-kill kinetics and bacterial protein estimation studies. Topographic images depicting synergism were obtained by scanning electron microscopy for Methicilin-resistant Staphylococcus aureus (MRSA) and Resistant Escherichia coli (R.E. coli). Results showed the presence of maximum phenolic (28.4 ± 0.67 µg GAE/mg extract) and flavonoid (11 ± 0.42 µg QE/mg extract) contents in MeOH extract. RP-HPLC results also displayed maximum polyphenols in MeOH extract followed by E.A extract. Clinical strains were resistant to cefixime whereas these were moderately inhibited by all extracts (MIC 150-300 µg/ml) except Aq. extract. E.A and n-Hex extracts demonstrated maximum synergism (Fractional inhibitory concentration index (FICI) 0.31) against R.E. coli. The n-Hex extract displayed total synergism against R.P. a with a 4-fold reduction in cefixime dose. Time-kill kinetics showed maximum inhibition of gram-negative bacterial growth from 3 to 12 h when treated at FICI and 2FICI values with > 10-fold reduction of the extracts' dose. All combinations demonstrate > 70 % protein content inhibition with bacterial cell wall disruption in SEM images. Fortunately, FICI concentrations have low hemolytic potential (<5%). Conclusively, F. indica extracts can mitigate antimicrobial resistance against cefixime and can be investigated in detail by in vivo and mechanistic studies.

Antibacterial potential and synergistic interaction between natural polyphenolic extracts and synthetic antibiotic on clinical isolates.

Emergence of antimicrobial resistance complicates treatment of infections by antibiotics. This has driven research on novel and combination antibacterial therapies. The present study evaluated synergistic antimicrobial activity of plant extracts and cefixime in resistant clinical isolates. Preliminary susceptibility profiling of antibiotics and antibacterial activity of extracts was done by disc diffusion and microbroth dilution assays. Checker-board, time-kill kinetics and protein content studies were performed to validate synergistic antibacterial activity. Results showed noteworthy quantities of gallic acid (0.24-19.7 µg/mg), quercetin (1.57-18.44 µg/mg) and cinnamic acid (0.02-5.93 µg/mg) in extracts of plants assessed by reverse-phase high performance liquid chromatography (RP-HPLC). Gram-positive (4/6) and Gram-negative (13/16) clinical isolates were intermediately susceptible or resistant to cefixime, which was used for synergistic studies. EA and M extracts of plants exhibited total synergy, partial synergy and indifferent characteristics whereas aqueous extracts did not show synergistic patterns. Time-kill kinetic studies showed that synergism was both time and concentration-dependent (2-8-fold decrease in concentration). Bacterial isolates treated with combinations at fractional inhibitory concentration index (FICI) showed significantly reduced bacterial growth, as well as protein content (5-62 %) as compared to extracts/cefixime alone treated isolates. This study acknowledges the selected crude extracts as adjuvants to antibiotics to treat resistant bacterial infections.

Characterization and comparative evaluation of wound healing potential of Ajugarin I and Ajuga bracteosa Wall. ex Benth.

Ajuga bracteosa (family: Lamiaceae), commonly known as kauri booti, is an important ethnomedicinal plant. The current research was conducted to appraise and compare the in vitro antioxidant and antibacterial profiles as well as in vivo wound healing potentials of Ajugarin I and A. bracteosa extract. Ajugarin I and polyphenols in A. bracteosa were enumerated by reversed-phase high-performance liquid chromatography analysis that confirmed significant amounts of Ajugarin I (2.2 ± 0.02 µg/mg DW) and other phenolic compounds (14 out of 17 standards). A. bracteosa (374.4 ± 0.20 µg AAE/mg of DW, 201.9 ± 0.20 µg AAE/mg of DW, 87 ± 0.30%) showed a higher antioxidant profile compared to Ajugarin I (221.8 ± 0.50 µg AAE/mg of DW, 51.8 ± 0.40 µg AAE/mg of DW, 27.65 ± 0.80%) with 1.86-, 3.89-, and 3.15-fold greater activity in ferric reducing antioxidant power, total antioxidant capacity, and free radical scavenging assays, respectively. Likewise, A. bracteosa showed antibacterial activity against 3/5 strains (MIC 25-200 µg/ml) than Ajugarin I (2/5 strains; MIC 50-200 µg/ml). Hemolytic (<2% hemolysis) and dermal toxicity tests rendered both samples non-toxic. Additionally, A. bracteosa (100 ± 2.34% at day 12; 9.33 ± 0.47 days) demonstrated 1.11- and 1.24-fold higher percent wound contraction and epithelization time, respectively, than Ajugarin I (95.6 ± 1.52% at day 12; 11.6 ± 0.47 days) as assessed by an excision wound model in mice. Histopathological examination further reinforced the better wound healing potential of A. bracteosa with good epithelization, collagen synthesis, fibroblast proliferation, and revascularization. Briefly, we endorse the significant comparative antioxidant, antibacterial, and wound healing activities of A. bracteosa and Ajugarin I and present these as prospective candidates for wound healing drugs.

Phytochemical analysis and comprehensive evaluation of pharmacological potential of Artemisia brevifolia Wall. ex DC.

Multitude of diseases and side effects from conventional drugs have surged the use of herbal remedies. Thus, the current study aimed to appraise various pharmacological attributes of Artemisia brevifolia Wall. ex DC. Extracts prepared by successive solvent extraction were subjected to phytochemical and multimode antioxidant assays. Various polyphenolics and artemisinin derivatives were detected and quantified using RP-HPLC analysis. Compounds present in methanol (M) and distilled water (DW) extracts were identified using high resolution mass spectrometry (HRMS). Extracts were pharmacologically evaluated for their antibacterial, antifungal, antimalarial, antileishmanial and antidiabetic potentials. Moreover, cytotoxicity against Artemiasalina, human cancer cell lines and isolated lymphocytes was assessed. Genotoxicity was evaluated using comet, micronucleus and chromosomal aberration assays. Lastly, anti-inflammatory potential was determined through a series of in vitro and in vivo assays using BALB/c mice. Maximum extract recovery (5.95% w/w) was obtained by DW extract. Highest phenolics and flavonoids content, total antioxidant capacity, total reduction potential, percentfree radical scavenging, ß-carotene scavenging and iron chelating activities were exhibited by M extract. RP-HPLC analysis revealed significant amounts of various polyphenolic compounds (vanillic acid, syringic acid, emodin and luteolin), artemisinin, dihydro artemisinin, artesunate and artemether in ethyl acetate (EA) extract. Total 40 compounds were detected through HRMS. A noteworthy antimicrobial activity (MIC 22.22 µg/ml) was exhibited by EA extract against A. fumigatus and several bacterial strains. Maximum antimalarial, antileishmanial, brine shrimp lethality and cytotoxic potential against cancer cells was manifested by EA extract. None of the extracts exhibited genotoxicity and toxicity against isolated lymphocytes. Highest α-amylase and α-glucosidase inhibition capacities were demonstrated by DW extract. Various in-vivo anti-inflammatory models revealed significant (p < 0.05) anti-inflammatory potential of M and DW extracts. In conclusion, present findings divulged theremarkable pharmacological potential of A. brevifolia and endorse its richness in artemisinin.

Appraisal of a novel extraction technique for estimation of cadmium content in pea seedlings based on human health risk assessment.

In this study, a novel extraction and safety evaluation method for heavy metals based on different functions of plants was proposed, and an edible plant (pea) was used as the research material to explore the feasibility of the novel method. Pea sprouts were cultured in cadmium (Cd) concentrations of 0, 1.0, 3.0, and 5.0 mg L-1, respectively. The Cd in pea sprouts was continuously extracted with 100 °C distilled water, 60% ethanol, 6% acetic acid, and simulated gastric juice. It was observed that highest amount of Cd (48.65-58.87%) was found in the extraction of roots with 6% acetic acid, followed by 100 °C distilled water (28.68-37.61%). While in stems, most of the Cd (70.73-85.39%) was extracted by 6% acetic acid. The recovery rate of the sequential chemical extraction technique employed in this experiment was between 93 and 106%. Compared with traditional methods, this study has its development potential in two aspects. First, it can determine which steps of sequential extractions of heavy metals in plants are the most harmful to humans. Secondly, corresponding measures can be taken to reduce heavy metals in vegetables used daily, such as soaking edible vegetables in vinegar for a short time. Novelty statement: In this study, a novel extraction and safety evaluation method for heavy metals based on different functions of plants was proposed, and an edible plant (pea) was used as the research material to explore the feasibility of the novel method. Compared with the commonly used extraction methods, the novel method is more reasonable and has greater development potential.

Efficient removal of acid orange 7 using a porous adsorbent-supported zero-valent iron as a synergistic catalyst in advanced oxidation process.

This study focuses on the synthesis of granular red mud reinforced by zero-valent iron (Fe@GRM) and its application for the removal acid orange 7 (AO7) from aqueous solution. Then ZVI is employed as a catalyst for the activation of persulfate (PS) to produce sulfate radicals (SO4•-) that are produced at 900 °C in an anoxic atmosphere using the direct reduction of iron oxide in the red mud with maize straw as the reductant. Furthermore, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) are used to illustrate the morphology and porous structure of the Fe@GRM. The X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) revealed that Fe@GRM was loaded with zero-valent iron. This characterization confirmed that the Fe@GRM was a porous structure material that contained zero-valent iron. The influence of conditions for AO7 elimination, including initial pH, Fe@GRM dosage, initial AO7 concentrations, and temperature, is also investigated. The removal efficiency of AO7 was 90.78% using Fe@GRM/PS, while only 18.15% was removed when Fe@GRM was used alone. The degradation kinetics were well fitted to a pseudo-first-order kinetic model, and the rate of removal increased with temperature, demonstrating an endothermic elimination process. The Arrhenius activation energy of the process was 20.77 kJ/mol, which indicated that the reduction of AO7 was a diffusion-mediated reaction. Fe@GRM is a low-cost material that demonstrated outstanding performance with great potential for wastewater treatment.

Isolation, identification, and characterization of diesel-oil-degrading bacterial strains indigenous to Changqing oil field, China.

In the present study, 12 indigenous diesel-oil-degrading bacteria were isolated from the petroleum-contaminated soils of the Changqing oil field (Xi'an, China). Measurement of the diesel-oil degradation rates of these strains by the gravimetric method revealed that they ranged from 42% to 66% within 2 weeks. The highest degradation rates were observed from strains CQ8-1 (66%), CQ8-2 (62.6%), and CQ11 (59%), which were identified as Bacillus thuringiensis, Ochrobactrum anthropi, and Bordetella bronchialis, respectively, based on their 16S rDNA sequences. Moreover, the physiological and biochemical properties of these three strains were analyzed by Gram staining, catalase, oxidase, and Voges-Proskauer tests. Transmission electron microscopy showed that all three strains were rod shaped with flagella. Gas chromatography and mass spectrometric analyses indicated that medium- and long-chain n-alkanes in diesel oil (C11-C29) were degraded to different degrees by B. thuringiensis, O. anthropi, and B. bronchialis, and the degradation rates gradually decreased as the carbon numbers increased. Overall, the results of this study indicate strains CQ8-1, CQ8-2, and CQ11 might be useful for environmentally friendly and cost-effective bioremediation of oil-contaminated soils.

Isolation and characterization of biosurfactant-producing and diesel oil degrading Pseudomonas sp. CQ2 from Changqing oil field, China.

In the present research investigation, 13 indigenous bacteria (from CQ1 to CQ13) were isolated from soil collected from Changqing oil field of Xi'an, China. Four promising biosurfactant producers (CQ1, CQ2, CQ4, and CQ13) were selected through primary screening among these 13 strains, including via drop collapse and oil-spreading methods. However, only the strain CQ2 showed the best biosurfactant production and was further screened by hemolytic assay, cetyl trimethyl ammonium bromide (CTAB), surface tension and emulsifying activity. The bacterium CQ2 has the ability to produce about 3.015 g L-1 of biosurfactant using glucose as the sole carbon source without any optimization. The produced biosurfactant could greatly reduce surface tension from 72.66 to 24.72 mN m-1 with a critical micelle concentration (CMC) of 30 mg L-1 and emulsify diesel oil up to 60.1%. The cell-free broth was found to be stable in wide temperature (4-100 °C), pH (6-12) and salinity (2-20%) ranges for surface and emulsifying activity. This biosurfactant was preliminarily found to be of a glycolipid nature as evident from thin-layer chromatographic (TLC) and Fourier transform infra-red spectroscopic (FTIR) analyses. Moreover, CQ2 was able to degrade 54.7% of diesel oil, which surprisingly could form a substantial amount of bioflocculants during the degradation process. Furthermore, the 16S rDNA sequence using the Genbank BLAST tool revealed that isolated CQ2 was closely related to species of Pseudomonas genus and, thus, was entitled Pseudomonas sp. CQ2. The results of residual diesel oil contents measured by GC-MS showed that C7-C28 hydrocarbons could be degraded by Pseudomonas sp. CQ2. Thus, these findings revealed that CQ2 could be applied for remediation of diesel oil/petroleum-contaminated waters and soils on a large scale.

Isolation of anticancer and antimicrobial metabolites from Epicoccum nigrum; endophyte of Ferula sumbul.

Owing to the importance of endophytes, current research was aimed to purify the secondary metabolites from targeted source. Ferula sumbul, a lipophilic extract of the endophyte was prepared in 10% methanol and partitioned with ethyl acetate and bioassay guided isolation was carried using standard protocols against bacterial, fungal and cancer cells. The active fractions consisted of three new metabolites (2-methyl-3-nonyl prodiginine, Bis (2-ethylhexyl) phthalate, and a meroterpenoid, Preaustinoid A). Their structures were confirmed with LCMS/MS. The purified metabolites showed valuable results against tested activities which concluded that these compounds have great potential and these may be applicable to textile (dyeing), pharmaceutical (drug, infectious agents) and food (preservatives) industries. This study reveals the potential of E. nigrum as an important source of bioactive compounds including 2-methyl-3-nonyl prodiginine, Bis (2-ethylhexyl) phthalate, and Preaustinoid A. This is first report of isolation of prodiginines as well as meroterpenoid and Bis (2-ethylhexyl) phthalate from Epicoccum nigrum.

Bacterial succession and degradative changes by biofilm on plastic medium for wastewater treatment.

Biofilms contain a diverse range of microorganisms and their varying extracellular polysaccharides. The present study has revealed biofilm succession associated with degradative effects on plastic (polypropylene) and contaminants in sludge. The wet weight of biofilm significantly (p < 0.05) increased; from 0.23 ± 0.01 to 0.44 ± 0.01 g. Similarly, the dry weight of the biofilm increased from 0.02 to 0.05 g. Significant reduction in pathogens (E. coli and feacal coliforms) by MPN technique (>80%) and in chemical parameters (decrease in COD, BOD5 of 73.32 and 69.94%) representing diminution of organic pollutants. Energy dispersive X-ray spectroscopy (EDS) of plastic revealed carbon and oxygen contents, further surface analysis of plastic by scanning electron microscopy (SEM) revealed emergence of profound bacterial growth on the surface. Fourier transform infrared (FTIR) spectroscopy conforms its biotransformation under aerobic conditions after 8 weeks. New peaks developed at the region 1050 and 969 cm(-1) indicating CO and CC bond formation. Thus plastic with 6 weeks old aerobic biofilm (free of pathogens, max. weight, and OD, efficient COD & BOD removal ability) is suggested to be maintained in fixed biofilm reactors for wastewater treatment.