Antibacterial and bacteriostatic potential of coelomic fluid and body paste of Pheretima posthuma (Vaillant, 1868) (Clitellata, Megascolecidae) against ampicillin resistant clinical bacterial isolates

Pheretima posthuma (Vaillant, 1868), a native earthworm of Pakistan and Southeast Asia, has wide utilization in vermicomposting and bioremediation process. In this study, P. posthuma coelomic fluid (PCF) and body paste (PBP) was evaluated as antibacterial agent against ampicillin (AMP) resistant five Gram positive and four Gram negative clinical isolates. The antibacterial effect of different doses (i.e. 25-100 µg/ml) of PCF and PBP along with AMP and azithromycin (AZM) (negative and positive controls, respectively) were observed through disc diffusion and micro-dilution methods. All nine clinical isolates were noticed as AMP resistant and AZM sensitive. Antibacterial effects of PCF and PBP were dose dependent and zone of inhibitions (ZI) against all clinical isolates were between 23.4 ± 0.92 to 0 ± 00 mm. The sensitivity profile of PCF and PBP against clinical isolates was noticed as 44.44 and 55.56%, respectively. Both PCF and PBP showed bacteriostatic (BTS) action against S. aureus, S. pyogenes, K. pneumonia, N. gonorrhoeae. Moreover, the cumulative BTS potential of PCF and PBP against all isolates was 66.67 and 55.56%, respectively. The MICs of PCF and PBP were ranged from 50-200 µg/ml against selected isolates. The bacterial growth curves indicated that PCF and PBP inhibited the growth of all isolates at their specific MIC concentrations. However, PBP has better antibacterial potential compared to PCF against selected isolates. Therefore, it is concluded that both PCF and PBP of P. posthuma possess antibacterial and BTS potential against ampicillin resistant clinical isolates. This organism might be considered as a second choice of antibacterial agents and can further be utilized in pharmaceutical industries for novel drug manufacturing by prospecting bioactive potential agents.

Synthesis, Molecular Modeling, DNA Damage Interaction, and Antioxidant Potential of Hesperidin Loaded on Gold Nanoparticles

The flavonoglycone hesperidin is recognized as a potent anti-inflammatory, anticancer, and antioxidant agent. However, its poor bioavailability is a crucial bottleneck regarding its therapeutic activity. Gold nanoparticles are widely used in drug delivery because of its unique properties that differ from bulk metal. Hesperidin loaded gold nanoparticles were successfully prepared to enhance its stability and bioactive potential, as well as to minimize the problems associated with its absorption. The free radical scavenging activities of hesperidin, gold nanoparticles, and hesperidin loaded gold nanoparticles were compared with that of Vitamin C and subsequently evaluated in vitro using 2,2-diphenyl-1-picrylhydrazyl assay. The antioxidant pharmacophore-based structure-activity relationship analysis was assessed by the density functional theory as well as quantum chemical calculations. Moreover, the structural properties were utilized using Becke’s three-parameter hybrid exchange and Lee-Yang-Parr’s correction of functional approaches. Hesperidin-loaded gold nanoparticles were found to decrease hydrogen peroxide (H2O2) and thus induce Deoxyribonucleic acid (DNA) instability. In addition, hesperidin-gold nanoparticles were observed to display important antioxidant potential as well as ameliorate the functional activity of macrophages against Escherichia coli, possibly protecting DNA. These particles might be appropriate for clinical trials and could prove useful for the treatment of various life-threatening disorders.

Dracunculin Inhibits Adipogenesis in Human Bone Marrow-Derived Mesenchymal Stromal Cells by Activating AMPK and Wnt/β-Catenin Signaling

Increased bone marrow adiposity is widely observed in patients with obesity and osteoporosis and reported to have deleterious effects on bone formation. Dracunculin (DCC) is a coumarin isolated from Artemisia spp. but, until now, has not been studied for its bioactive potential except antitrypanosomal activity. In this context, current study has reported the anti-adipogenic effect of DCC in human bone marrow-derived mesenchymal stromal cells (hBM-MSCs). DCC dose-dependently inhibited the lipid accumulation and expression of adipogenic transcription factors peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα) in hBM-MSCs induced to undergo adipogenesis. To elucidate its action mechanism, the effect of DCC on Wnt/β-catenin and AMPK pathways was examined. Results showed that DCC treatment activated Wnt/β-catenin signaling pathway via AMPK evidenced by increased levels of AMPK phosphorylation and Wnt10b expression after DCC treatment. In addition, DCC treated adipo-induced hBM-MSCs exhibited significantly increased nuclear levels of β-catenin compared with diminished nuclear PPARγ levels. In conclusion, DCC was shown to be able to hinder adipogenesis by activating the β-catenin via AMPK, providing potential utilization of DCC as a nutraceutical against bone marrow adiposity.

Physicochemical characterization of C-phycocyanin from Plectonema sp. and elucidation of its bioactive potential through in silico approach

C-phycocyanin (C-PC), the integral blue-green algae (BGA) constituent has been substantially delineated for its biological attributes. Numerous reports have illustrated differential extraction and purification techniques for C-PC, however, there exists paucity in a broadly accepted process of its isolation. In the present study, we reported a highly selective C-PC purification and characterization method from nontoxic, filamentous and non-heterocystous cyanobacterium Plectonema sp. C-PC was extracted by freeze-thawing, desalted and purified using ion-exchange chromatography. The purity of C-PC along with its concentration was found to be 4.12 and 245 µg/ml respectively. Comparative characterization of standard and purified C-PC was performed using diverse spectroscopic techniques namely Ultra Violet-visible, fluorescence spectroscopy and Fourier transform infrared (FT-IR). Sharp peaks at 620 nm and 350 nm with UV-visible and FT-IR spectroscopy respectively, confirmed amide I bands at around 1638 cm-1 (C=O stretching) whereas circular dichroism (CD) spectra exhibited α-helix content of secondary structure of standard 80.59% and 84.59% of column purified C-PC. SDS-PAGE exhibited two bands of α and β subunits 17 and 19 kDa respectively. HPLC evaluation of purified C-PC also indicated a close resemblance of retention peak time (1.465 min, 1.234 min, 1.097 min and 0.905 min) with standard C-PC having retention peak timing of 1.448 min, 1.233 min and 0.925 min. As a cautious approach, the purified C-PC was further lyophilized to extend its shelf life as compared to its liquid isoform. To evaluate the bioactive potential of the purified C-PC in silico approach was attempted. The molecular docking technique was carried out of C-PC as a ligand-protein with free radicals and α-amylase, α-glucosidase, glycogen synthase kinase-3 and glycogen phosphorylase enzymes as receptors to predict the free radical scavenging (antioxidant) and to target antidiabetic property of C-PC. In both receptors free radicals and enzymes, ligand C-PC plays an important role in establishing interactions within the cavity of active sites. These results established the antioxidant potential of C-PC and also give a clue towards its antidiabetic potential warranting further research.

GC-MS Profiling and Antifungal Activity of Secondary Metabolite from Endophytic Fungus of Giloy

The endophytic microbiota is considered to be one of the consistent and noble souce of potential and unique natural amalgams. These natural amalgams carry diverse pharmaceutical significance which the reason for their importance among research fields. The diversity of plants carries much more diversity of the endophytes as their mutual parts where both are benefited from each other. The current work deals with the isolation of the endophytic fungus from Tinospora cordifolia, for which the leaves were used after the surface sterilization, followed by the production of secondary metabolite by the endophytic isolates through submerged fermentation technique. The produced metabolite was extracted by liquid-liquid extraction technique, which was further used for evaluating its antifungal potential against Candida albicans and the obtained results show their considerable potential. The GC-MS profiling of secondary metabolite was conducted to determine the presence of some bioactive compounds in them, and as a result, some potential compounds detected are Levoglucosenone, Silanediol, Nonane, D-Allose, 5-Hydroxymethylfurfural. Since these compounds are biologically important in various aspects which gives the diversified application to the secondary metabolites. The study concludes the potential of secondary metabolites from endophytic fungus of Tinospora cordifolia and further investigation can be approached on determining the same from other plants, and also evaluating another bioactive potential of secondary metabolites.

Influence of Natural Polysaccharides on Properties of the Biomicroconcrete-Type Bioceramics

In this paper, novel hybrid biomicroconcrete-type composites were developed and investigated. The solid phase of materials consisted of a highly reactive α -tricalcium phosphate (α-TCP) powder, hybrid hydroxyapatite-chitosan (Hap-CTS) material in the form of powder and granules (as aggregates), and the polysaccharides sodium alginate (SA) or hydroxypropyl methylcellulose (HPMC). The liquid/gel phase in the studied materials constituted a citrus pectin gel. The influence of SA or HPMC on the setting reaction, microstructure, mechanical as well as biological properties of biomicroconcretes was investigated. Studies revealed that manufactured cement pastes were characterized by high plasticity and cohesion. The dual setting system of developed biomicroconcretes, achieved through α-TCP setting reaction and polymer crosslinking, resulted in a higher compressive strength. Material with the highest content of sodium alginate possessed the highest mechanical strength (~17 MPa), whereas the addition of hydroxypropyl methylcellulose led to a subtle compressive strength decrease. The obtained biomicroconcretes were chemically stable and characterized by a high bioactive potential. The novel biomaterials with favorable physicochemical and biological properties can be prosperous materials for filling bone tissue defects of any shape and size.

Phenolic Composition and Biological Properties of Cynara cardunculus L. var. altilis Petioles: Influence of the Maturity Stage

Hydroethanolic extracts of cardoon petioles collected at sixteen growth stages (P1–P16) were characterized in terms of their phenolic composition and bioactive potential (antioxidant, cytotoxic, anti-inflammatory, and antimicrobial activities). Fifteen phenolic compounds were tentatively identified (i.e., ten phenolic acids and five flavonoid glycosides); the main compounds were 5-O-caffeoylquinic and 1,5-di-O-caffeoylquinic acids. Samples collected at early maturity (P1–P4) presented a weak positive correlation between the higher content in polyphenols (P3: 101-mg/g extract) and better inhibition capacity against thiobarbituric acid reactive substance formation (TBARS; P3: IC50 = 5.0 µg/mL). Samples at intermediate maturation stages (P9) presented higher cytotoxic and anti-inflammatory potential. Moreover, immature petioles showed greater antihemolytic (OxHLIA; P4: IC50 = 65 and 180 µg/mL for Δt of 60 and 120 min, respectively) and antibacterial activity. The antifungal activity varied depending on the maturation stage and the fungi strain. In conclusion, the maturation stage may greatly affect the polyphenols composition and content and the bioactive potential of cardoon petioles.