Enhancement of shelf-life of food items via immobilized enzyme nanoparticles on varied supports. A sustainable approach towards food safety and sustainability.

This study was designed to extend the shelf life of fruits and vegetables through a novel technique based on utilization of microbially driven enzyme glucose oxidase and casting a fine layer of hydrogen peroxide on the food item that protected the fruit from decay. The produced nanoparticles (ZnO, Ag) were ligated with Glucose Oxidize (GOx) purified from Aspergillus niger. Post ligation studies revealed that ligated enzymes display relatively enhanced activity. Four types of sprays were prepared in order to compare their effectiveness. Glucose oxidase/silver nanoparticles (GOx/AgNPs), glucose oxidase/zinc oxide nanoparticles (GOx/ZnONPs), AgNPs and ZnONPs sprays were applied to guava fruit samples as post-harvest therapeutic agents for a period of 15 days. Fruit quality parameters such as total suspended solids (TSS), pH, weight loss, DPPH free radical capturing performance and firmness confirms that usage of the bioconjugates especially that of GOx/ZnONP was curiously active to maintain the physical appearance of fruit well along with no such deterioration in chemical composition of fruit. Consequently, enzymes ligated on the surface of nanoparticles (ZnONP) are exceptional for extension of post-harvest shelf life of fruits such as guava.

Biomimetic nanoplasmonic sensor for rapid evaluation of neutralizing SARS-CoV-2 monoclonal antibodies as antiviral therapy.

Monoclonal antibody (mAb) therapy is one of the most promising immunotherapies that have shown the potential to prevent or neutralize the effects of COVID-19 in patients at very early stages, with a few formulations recently approved by the European and American medicine agencies. However, a main bottleneck for their general implementation resides in the time-consuming, laborious, and highly-specialized techniques employed for the manufacturing and assessing of these therapies, excessively increasing their prices and delaying their administration to the patients. We propose a biomimetic nanoplasmonic biosensor as a novel analytical technique for the screening and evaluation of COVID-19 mAb therapies in a simpler, faster, and reliable manner. By creating an artificial cell membrane on the plasmonic sensor surface, our label-free sensing approach enables real-time monitoring of virus-cell interactions as well as direct analysis of antibody blocking effects in only 15 min assay time. We have achieved detection limits in the 102 TCID50/mL range for the study of SARS-CoV-2 viruses, which allows to perform neutralization assays by only employing a low-volume sample with common viral loads. We have demonstrated the accuracy of the biosensor for the evaluation of two different neutralizing antibodies targeting both Delta and Omicron variants of SARS-CoV-2, with half maximal inhibitory concentrations (IC50) determined in the ng/mL range. Our user-friendly and reliable technology could be employed in biomedical and pharmaceutical laboratories to accelerate, cheapen, and simplify the development of effective immunotherapies for COVID-19 and other serious infectious diseases or cancer.

Fluorene-Based Fluorometric and Colorimetric Conjugated Polymers for Sensitive Detection of 2,4,6-Trinitrophenol Explosive in Aqueous Medium.

Nitroaromatic explosives are a class of compounds that are responsible for various health hazards and terrorist outrages. Among these, sensitive detection of 2,4,6-trinitrophenol (TNP) explosive has always been highly desirable considering public health and national security. In this regard, three fluorene-based conjugated polymers (CP 1, CP 2, and CP 3) were synthesized through the Suzuki-Miyaura coupling reaction and were found to be highly sensitive for fluorescence detection of TNP with detection limits of 3.2, 5.7, and 6.1 pM, respectively. Excellent selectivity of CPs toward TNP was attributed to their unique π-π interactions based on fluorescence studies and density functional theory (DFT) calculations. The high sensitivity of CPs to TNP was attributed to the static quenching mechanism based on the photoinduced electron transfer process and was evaluated by fluorescence, UV-visible absorption, dynamic light scattering, Job's plots, the Benesi-Hildebrand plots, and DFT calculations. CPs were also used for colorimetric and real-water sample analysis for the detection of TNP explosive. Meanwhile, sensor-coated test strips were fabricated for on-site detection of TNP, which makes them convenient solid-supported sensors.

Postharvest shelf life enhancement of peach fruit treated with glucose oxidase immobilized on ZnO nanoparticles.

For the shelf life extension of fruits, we envisioned a novel antimicrobial approach that is based on the production of a thin layer of hydrogen peroxide at the surface of food by utilizing the bioactivity of glucose oxidase (GOx). The enzyme, purified from Aspergillus Niger, was immobilized on zinc oxide nanoparticles and then suspended in a buffer to prepare a spraying solution of GOx/ZnONPs. Post-immobilization analyses indicated that immobilized enzyme showed higher activity as compared to the free enzyme. The GOx/ZnONPs spray was applied for postharvest treatment of peach. The control and treatment groups were stored at ambient conditions for fifteen days and standard quality parameters were analyzed. In contrast to the control group, the GOx/ZnONPs spray treatment was remarkably effective in maintaining the physiological appearance of fruits even more than 12 days and showed a significant reduction in the decrease of weight, firmness, TSS, and DPPH free radical scavenging activity of fruits. Thus GOx/ZnONPs is an excellent platform to extend the postharvest shelf life of peach.

Synthesis, evaluation of thymidine phosphorylase and angiogenic inhibitory potential of ciprofloxacin analogues: Repositioning of ciprofloxacin from antibiotic to future anticancer drugs.

Over expression of thymidine phosphorylase (TP) in various human tumors compared to normal healthy tissue is associated with progression of cancer and proliferation. The 2-deoxy-d-ribose is the final product of thymidine phosphorylase (TP) catalyzed reaction. Both TP and 2-deoxy-d-ribose are known to promote unwanted angiogenesis in cancerous cells. Discovery of potent inhibitors of thymidine phosphorylase (TP) can offer appropriate approach in cancer treatment. A series of ciprofloxacin 2, 3a-3c, 4a-4d, 5a-5b, 6 and 7 has been synthesized and characterized using spectroscopic techniques. Afterwards, inhibitory potential of synthesized ciprofloxacin 2, 3a-3c, 4a-4d, 5a-5b, 6 and 7 against thymidine phosphorylase enzyme was assessed. Out of these twelve analogs of ciprofloxacin nine analogues 3a-3c, 4a-4c, 5a-5b and 6 showed good inhibitory activity against thymidine phosphorylase. Inhibitory activity as presented by their IC50 values was found in the range of 39.71 ± 1.13 to 161.89 ± 0.95 µM. The 7-deazaxanthine was used as a standard inhibitor with IC50 = 37.82 ± 0.93 µM. Furthermore, the chick chorionic allantoic membrane (CAM) assay was used to investigate anti-angiogenic activity of the most active ciprofloxacin-based inhibitor 3b. To enlighten the important binding interactions of ciprofloxacin derivatives with target enzyme, the structure activity relationship and molecular docking studies of chosen ciprofloxacin analogues was discussed. Docking studies revealed key π-π stacking, π-cation and hydrogen bonding interactions of ciprofloxacin analogues with active site residues of thymidine phosphorylase enzyme.

Thyroxine-loaded chitosan/carboxymethyl cellulose/hydroxyapatite hydrogels enhance angiogenesis in in-ovo experiments.

Angiogenesis is one of the most important processes in repair and regeneration of many tissues and organs. Blood vessel formation also play a major role in repair of dental tissue(s) after ailments like periodontitis. Here we report the preparation of chitosan/carboxymethyl cellulose/hydroxyapatite based hydrogels, loaded with variable concentrations of thyroxin i.e., 0.1 µg/ml, 0.5 µg/ml and 1 µg/ml. Scanning electron microcopy images (SEM) showed all hydrogels were found to be porous and solution absorption study exhibited high swelling potential in aqueous media. FTIR spectra confirmed that the used materials did not change their chemical identity in synthesized hydrogels. The synthesized hydrogels demonstrated good bending, folding, rolling and stretching abilities. The hydrogels were tested in chick chorioallantoic membrane (CAM) assay to investigate their angiogenic potential. Hydrogel containing 0.1 µg/ml of thyroxine showed maximum neovascularization. For cytotoxicity analyses, preosteoblast cells (MC3T3-E1) were seeded on these hydrogels and materials were found to be non-toxic. These hydrogels with pro-angiogenic activity possess great potential to be used for periodontal regeneration.

A nanocomposite prepared from magnetite nanoparticles, polyaniline and carboxy-modified graphene oxide for non-enzymatic sensing of glucose.

The authors report on the synthesis of carboxy functionalized graphene oxide (fGO) decorated with magnetite (Fe3O4) nanoparticles. The resulting nanomaterial was used to prepare a composite with polyaniline (PANI) which was characterized by UV-vis, Fourier transform-infrared and Raman spectroscopies. Its surface morphologies were characterized by atomic force and scanning electron microscopies. A screen-printed carbon electrode was then modified with the nanocomposite to obtain an enzyme-free glucose sensor. The large surface of fGO and Fe3O4 along with the enhanced charge transfer capability of PANI warrant a pronounced electrochemical response (typically measured at 0.18 V versus Ag/AgCl) which is suppressed in the presence of glucose. This reduction of current by glucose was used to design a sensitive method for quantification of glucose. The response of the modified SPCE is linear in the 0.05 µM - 5 mM glucose concentration range, and the lower detection limit is 0.01 µM. Graphical abstract Schematic illustration of in-situ anchoring of Iron oxide on functionalized graphene oxide and synthesis of its polymeric nanocomposite for non-enzymatic detection of Glucose. The nanocomposite modified screen printed interface enabled monitoring of glucose at lower potential with higher precision. GO (graphene oxide), fGO (functionalized graphene oxide), PANI (polyaniline).

A Review of the Construction of Nano-Hybrids for Electrochemical Biosensing of Glucose.

Continuous progress in the domain of nano and material science has led to modulation of the properties of nanomaterials in a controlled and desired fashion. In this sense, nanomaterials, including carbon-based materials, metals and metal oxides, and composite/hybrid materials have attracted extensive interest with regard to the construction of electrochemical biosensors. The modification of a working electrode with a combination of two or three nanomaterials in the form of nano-composite/nano-hybrids has revealed good results with very good reproducibility, stability, and improved sensitivity. This review paper is focused on discussing the possible constructs of nano-hybrids and their subsequent use in the construction of electrochemical glucose biosensors.