METHODOLOGY: E. histolytica HM-1:IMSS genomic DNA was isolated and two putative choline/ethanolamine kinase genes (EhCK1 and EhCK2) were cloned and expressed from Escherichia coli BL21 strain. Enzymatic characterizations were further carried out on the purified enzymes.
RESULTS: EhCK1 and EhCK2 were identified from E. histolytica genome. The deduced amino acid sequences were more identical to its homologues in human (35-48%) than other organisms. The proteins were clustered as ethanolamine kinase in the constructed phylogeny tree. Sequence analysis showed that they possessed all the conserved motifs in choline kinase family: ATP-binding loop, Brenner's phosphotransferase motif, and choline kinase motif. Here, the open reading frames were cloned, expressed, and purified to apparent homogeneity. EhCK1 showed activity with choline but not ethanolamine. The biochemical characterization showed that it had a Vmax of 1.9 ± 0.1 µmol/min/mg. Its Km for choline and ATP was 203 ± 26 µM and 3.1 ± 0.4 mM, respectively. In contrast, EhCK2 enzymatic activity was only detected when Mn2+ was used as the co-factor instead of Mg2+ like other choline/ethanolamine kinases. Highly sensitive and specific antibody against EhCK1 was developed and used to confirm the endogenous EhCK1 expression using immunoblotting.
CONCLUSIONS: With the understanding of EhC/EK importance in phospholipid metabolism and their unique characteristic, EhC/EK could be a potential target for future anti-amoebiasis study.
MATERIALS AND METHODS: The produced nanoparticles were characterized using UV-visible spectroscopy. The Bovine Serum Assay (BSA) and Egg Albumin (EA) assays were used to assess the anti-inflammatory effects. The protein denaturation of AgNPs was calculated and compared to a standard to determine the anti-inflammatory activity of green synthesized AgNPs. Using varying concentrations of OGmediated AgNPs in Mueller Hinton Agar (MHA), the antimicrobial effects of OG have been investigated against E. coli, S. aureus, and Pseudomonas sp. Additionally, by measuring optical density, the time-kill curve analysis for E. Coli and S. Aureus has been examined from one hour for up to five hours.
RESULTS: The green synthesized AgNPs were developed successfully using a plant Ocimum gratissimum. The synthesized AgNPs exhibited a maximum absorption peak at 440 nm and SEM analysis revealed that the synthesized AgNPs were spherical and oval. The result findings of the anti-inflammatory activity reveal that AgNPs have great potential when compared to the standard. At the concentration of 50 μg/mL, AgNPs exhibit 76% in BSA assay and 74% in EA assay, where the standard shows 80% inhibition. The antimicrobial activity showed a zone of inhibition around 19mm for E. coli and a 20mm zone of inhibition for S. aureus and Pseudomonas sp., which shows the efficacy of AgNPs. The time-kill assay shows that the optical density of E. coli and S. aureus was reduced to 0.1 after 5 hours of incubation, which shows the potential of green synthesized AgNPs.
CONCLUSION: OG-mediated AgNPs have both antiinflammatory and antimicrobial effects. Anti-inflammatory effects are better when compared to standard drugs. Antimicrobial effects are better for Gram-negative bacteria.
METHODS: Antimicrobial activity was determined with disc diffusion and broth microdilution assays against eight skin colonising microorganisms including Staphylococcus aureus, Staphylococcus epidermidis, Salmonella enterica, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia followed by further fractionation of the pods ethyl acetate fraction by column chromatography along with preparative thin-layer chromatography. Quantification of bacterial death mechanism was elucidated by the measurement of hole size in cell wall that has been induced by extract constituents via field-emission scanning electron microscopy (FESEM).
RESULTS: Four fractions showed significant antimicrobial activity against the six microorganisms tested (p
METHODS: The extracts were prepared and initially assessed for their phytochemical composition and antibacterial activity. Then, AgNPs were synthesized from these extracts at room temperature, and various analytical techniques, including UV-visible spectroscopy, X-ray diffraction (XRD), ATIR-FTIR, zeta sizer, scanning electron microscopy (SEM), and energy-dispersive X-ray analysis (EDXA), were used to characterize the NPs. After confirmation of prepared NPs, they were subjected to their antibacterial activity.
RESULTS: HPLC analysis demonstrated the presence of eight phytoconstituents in organic ginger extracts. The absorption spectra of the silver suspension exhibited surface plasmon resonance peaks with maxima between 420 and 448 nm. Functional groups like C-H, N-H, OH, C-O-C, C=O, and C-O were identified in both the organic and aqueous extracts of Z. officinale, playing a key role in the formation of AgNPs, as characterized by ATR-FTIR analysis. Both ginger organic and aqueous extract synthesized AgNPs crystalline structure was shown in XRD analysis and the particle size distribution showed average diameter of 200.5 nm of AgNPs from aqueous extracts. Scanning Electron Microscopy displayed spherical structure and EDA results showed the percentage of elements in synthesized AgNPs using plant extracts. Most promising antibacterial activity was obtained against Escherichia coli ie 20.83±0.53 for 100 µg/mL.
CONCLUSION: The results of the current study showed that AgNPs synthesized from different ginger extracts have promising antibacterial properties and can be potential candidates for alternative treatment options for bacterial infections.