A novel device for swift and efficient CD44 protein digestion of pipette tips in human serum.

For molecular diagnostics in modern biomedical research, electrospray ionisation mass spectrometry (ESI-MS) based on proteome profiling is important. Now a days, sample preparation such as proteolysis and protein extraction remain incredibly challenging and inefficient. Recent sample-preparation methods based on micro tips show promising results toward the aim "a proteome in an hour". Proteolysis at the tip, is still infrequently observed and does not represent the processing of complex bio-samples. In this study, we outline a unique technique for detecting and extracting human serum CD44 biomarkers by ligand-protein interactions. This method employs macropores silica particles (MPSP) or (MOSF) modified with hyaluronic acid (HA). In order to assist in the profile of the human serum proteome, we limitations of immunoassays for rapid and multimodal proteolysis. For effective in situ proteolysis, in micropipette tips, MPSP were designed as nanoreactors with variable pore size and surface chemistry. In MS-based bottom-up proteome analysis, the device as-built demonstrated favourable sensitivity (LOD of 0.304 ± 0.007 ng/mL and LOQ of 0.973 ± 0.054 ng/mL), selectivity, durability (at -20 °C for 2 months), reuse (at least 10 times), and minimal memory impact. In addition, we examined into specific surface chemistries of nanoparticles for the absorption of proteins in serum and profiled the HA-binding serum proteome, setting a new preliminary benchmark for future databases. Our study not only helped establish a new platform for extracting/detection of CD44 and identifying the HA-binding proteome, but it also offered design recommendations for ligand affinity-based techniques for the antibody-free study of serum biomarkers with a view towards diagnostic applications.

Biosynthesis and characterization of copper oxide nanoparticles from indigenous fungi and its effect of photothermolysis on human lung carcinoma.

In this report, copper oxide nanoparticles (TA-CuO NPs) were synthesized using cell-free extract of Trichoderma asperellum and assessed their photothermal induced anticancerous activity. The fungal mediated TA-CuO NPs was confirmed by the surface plasmon resonance at 285-295 nm. The amide (CO) and aromatic (CC) groups in secondary metabolites of the extract was found to be an encapsulating or reducing agents for TA-CuO NPs, as indicated by IR spectra. Crystalline nature by cubic face-centered structure of the TA-CuO NPs was confirmed by XRD and their size ranges from 10 to 190 nm and an average of 110 nm by particle size analyzer (PSA). The Ultra HRSEM study revealed spherical shaped TA-CuO NPs. The FETEM results were also in strong agreement with PSA and UHR SEM. The survey-scan spectrum of XPS indicated the presence of C1s (47.83%), Cu2p (16.11%), Na1s (2.2%) and O1s (33.86%). The cell death was significantly found higher in photothermal induced by near-infrared laser (TA-CuO NPs-NIR) treated than that of TA-CuO NPs treatment. The level of ROS (35.62%) was higher in the treated cells than that of the untreated control, in accordance with the nucleus damage and losses in the mitochondrial membrane potential (ΔΨm). The upregulation of Bcl-2 in the untreated cells and Cas-3 in TA-CuO NPs-NIR treated cells was confirmed by western blot analysis. This work agreed with the potential biogenic TA-CuO NPs for promising in vitro photothermolysis of cancer cells, for the development of anticancer nanotherapeutics.

Green synthesis and characterization of biologically active nanosilver from seed extract of Gardenia jasminoides Ellis.

This article reports the utilization of seed extract (GSE) from Gardenia jasminoides Ellis. in the synthesis of silver nanoparticles (Gs-AgNPs) with versatile biological activities. The synthesized Gs-AgNPs were spherical in shape, crystal lattice with an average size of 20 nm as confirmed by UV-vis spectrum, X-ray diffractometer (XRD), Transmission electron microscopy with Energy dispersive X-ray spectroscopy (TEM-EDS) and particle size analyses (PSA). Phenolic compounds, proteins, and terpenoids were likely involved in the Gs-AgNPs synthesis, as indicated by Fourier-transform infrared spectroscopy (FTIR) analysis. The minimum bactericidal concentration (MBC) of the Gs-AgNPs was 12.5 µg·ml-1 for S. enterica Typhimurium and 10 µg·ml-1 for S. aureus. The MBC of the Gs-AgNPs induced >70% bacterial cell death within 60 min, as confirmed by growth curve analysis followed by Confocal laser scanning microscope (CLSM). Gs-AgNPs showed the highest scavenging activity for 1, 2-diphenyl-1-picrylhydrazyl DPPH radical (92.3 ±â€¯0.86%), Nitric oxide (NO) radical (72.5 ±â€¯2.15%), and Hydrogen peroxide H2O2 radical (85.25 ±â€¯1.45%). Anticancer results revealed an IC50 of 15.625 ±â€¯1.3 µg·ml-1 for Gs-AgNPs, whereas it was 580.54 ±â€¯2.5 µg·ml-1 for GSE. The Gs-AgNPs generated high reactive oxygen species (ROS) resulting in induced apoptosis as evident by up-regulation of apoptosis-related protein. In addition, the photocatalytic results revealed about 92% of the reduction in Coomassie Brilliant Blue dye color with Gs-AgNPs. Hence, this work provided economically viable and ecologically sustainable Gs-AgNPs as an alternative biomaterial for future therapeutic applications as antimicrobial, antioxidant, anti-cancer agents and in dye degradation for water remediation.