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The synthesis of a Ni-doped ZnO nanocomposite incorporating chitosan (CS/Ni-doped ZnO) was achieved via a precipitation method, followed by annealing at 250 °C. This study comprehensively examined the nanocomposite's structural, functional, morphological, and porosity properties using various analytical techniques, including X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), high-resolution scanning electron microscopy (HR-SEM), transmission electron microscopy (TEM), and Brunauer–Emmett–Teller (BET) analysis. The presence of chitosan (CS) and nickel (Ni) within the nanocomposite, along with their influence on reducing the band gap of ZnO particles and enhancing the generation of electron-hole pairs, was confirmed using UV-visible near-infrared spectroscopy (UV-vis-NIR). The electrochemical properties of the CS/Ni-doped ZnO nanocomposite were investigated via electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) by utilizing a phosphate buffer solution with a pH of 6, which closely resembled the typical pH of bacterial cell walls. Finally, the prepared CS/Ni-doped ZnO nanocomposite was evaluated for its antibacterial and anticancer activities. The results demonstrated the highest inhibition of bacterial growth in P. vulgaris, whereas the lowest inhibition was found in S. aureus across various concentrations, thus highlighting its potential in antimicrobial applications. The cytotoxicity of CS/Ni-doped ZnO nanocomposites demonstrated remarkable effects with a half-maximum inhibitory concentration of approximately 80 ± 0.23 µg mL−1 against MCF-7 breast cancer cell lines, following a dose-dependent manner. [ABSTRACT FROM AUTHOR]

The seafood industry plays a huge role in the blue economy, exploiting the advantage of the enriched protein content of marine organisms such as shrimps and molluscs, which are cultured in aquafarms. Diseases greatly affect these aquatic organisms in culture and, hence, there is need to study, in detail, their innate immune mechanisms. Hemocyanin is a non-specific innate defense molecule present in the blood cells of several invertebrates, especially molluscs, arthropods, and annelids. It is concerned with oxygen transport, blood clotting, and immune enhancement. In the present study, this macromolecular metalloprotein was isolated from the hemolymph of the marine snail Hemifusus pugilinus (Born, 1778) using Sephadex G-100 gel filtration column chromatography. It occurred as a single band (MW 80 kDa) on SDS-PAGE. High-performance liquid chromatography (HPLC) of the purified hemocyanin showed a single peak with a retention time of 4.3 min. The secondary structure and stability of the protein were detected using circular dichroism (CD), and the spectra demonstrated negative ellipticity bands close to 208 nm and 225 nm, indicating β-sheets. Further exploration of the purified hemocyanin revealed remarkable antimicrobial and antibiofilm activities against Gram-positive (Enterococcus faecalis and Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa and Proteus vulgaris) at a concentration of 1–5 μg/mL. Spectrophotometric and in situ microscopic analyses (CLSM) unveiled the potential of the purified hemocyanin to inhibit biofilm formation in these bacteria with a minimal inhibitory concentration of 40 μg/mL. Furthermore, H. pugilinus hemocyanin (10 μg/mL concentration) displayed antifungal activity against Aspergillus niger. The purified hemocyanin was also assessed for cytotoxicity against human cancer cells using cell viability assays. Altogether, the present study shows that molluscan hemocyanin is a potential antimicrobial, antibiofilm, antifungal, anticancer, and immunomodulatory agent, with great scope for application in the enhancement of the immune system of molluscs, thereby facilitating their aquaculture. [ABSTRACT FROM AUTHOR]

Heat shock proteins (HSPs) encompass both extrinsic chaperones and stress proteins. These proteins, with molecular weights ranging from 14 to 120 kDa, are conserved across all living organisms and are expressed in response to stress. The upregulation of specific genes triggers the synthesis of HSPs, facilitated by the interaction between heat shock factors and gene promoter regions. Notably, HSPs function as chaperones or helper molecules in various cellular processes involving lipids and proteins, and their upregulation is not limited to heat-induced stress but also occurs in response to anoxia, acidosis, hypoxia, toxins, ischemia, protein breakdown, and microbial infection. HSPs play a vital role in regulating protein synthesis in cells. They assist in the folding and assembly of other cellular proteins, primarily through HSP families such as HSP70 and HSP90. Additionally, the process of the folding, translocation, and aggregation of proteins is governed by the dynamic partitioning facilitated by HSPs throughout the cell. Beyond their involvement in protein metabolism, HSPs also exert a significant influence on apoptosis, the immune system, and various characteristics of inflammation. The immunity of aquatic organisms, including shrimp, fish, and shellfish, relies heavily on the development of inflammation, as well as non-specific and specific immune responses to viral and bacterial infections. Recent advancements in aquatic research have demonstrated that the HSP levels in populations of fish, shrimp, and shellfish can be increased through non-traumatic means such as water or oral administration of HSP stimulants, exogenous HSPs, and heat induction. These methods have proven useful in reducing physical stress and trauma, while also facilitating sustainable husbandry practices such as vaccination and transportation, thereby offering health benefits. Hence, the present review discusses the importance of HSPs in different tissues in aquatic organisms (fish, shrimp), and their expression levels during pathogen invasion; this gives new insights into the significance of HSPs in invertebrates. [ABSTRACT FROM AUTHOR]