Genipin (GN) is a natural molecule extracted from the fruit of Gardenia jasminoides Ellis according to modern microbiological processes. Genipin is considered as a favorable cross-linking agent due to its low cytotoxicity compared to widely used cross-linkers; it cross-links compounds with primary amine groups such as proteins, collagen, and chitosan. Chitosan is a biocompatible polymer that is currently studied in bone tissue engineering for its capacity to promote growth and mineral-rich matrix deposition by osteoblasts in culture. In this work, two genipin cross-linked chitosan scaffolds for bone repair and regeneration were prepared with different GN concentrations, and their chemical, physical, and biological properties were explored. Scanning electron microscopy and mechanical tests revealed that nonremarkable changes in morphology, porosity, and mechanical strength of scaffolds are induced by increasing the cross-linking degree. Also, the degradation rate was shown to decrease while increasing the crosslinking degree, with the high cross-linking density of the scaffold disabling the hydrolysis activity. Finally, basic biocompatibility was investigated in vitro, by evaluating proliferation of two human-derived cell lines, namely, the MG63 (human immortalized osteosarcoma) and the hMSCs (human mesenchymal stem cells), as suitable cell models for bone tissue engineering applications of biomaterials.

Solute Carrier 15 (SLC15) family, alias H-coupled oligopeptide cotransporter family, is a group of membrane transporters known for their role in the cellular uptake of di- and tripeptides (di/tripeptides) and peptide-like molecules. Of its members, SLC15A1 (PEPT1) chiefly mediates intestinal absorption of luminal di/tripeptides from dietary protein digestion, while SLC15A2 (PEPT2) mainly allows renal tubular reabsorption of di/tripeptides from ultrafiltration, SLC15A3 (PHT2) and SLC15A4 (PHT1) possibly interact with di/tripeptides and histidine in certain immune cells, and SLC15A5 has unknown function. Our understanding of this family in vertebrates has steadily increased, also due to the surge of genomic-to-functional information from 'non-conventional' animal models, livestock, poultry, and aquaculture fish species. Here, we review the literature on the SLC15 transporters in teleost fish with emphasis on SLC15A1 (PEPT1), one of the solute carriers better studied amongst teleost fish because of its relevance in animal nutrition. We report on the operativity of the transporter, the molecular diversity, and multiplicity of structural-functional solutions of the teleost fish orthologs with respect to higher vertebrates, its relevance at the intersection of the alimentary and osmoregulative functions of the gut, its response under various physiological states and dietary solicitations, and its possible involvement in examples of total body plasticity, such as growth and compensatory growth. By a comparative approach, we also review the few studies in teleost fish on SLC15A2 (PEPT2), SLC15A4 (PHT1), and SLC15A3 (PHT2). By representing the contribution of teleost fish to the knowledge of the physiology of di/tripeptide transport and transporters, we aim to fill the gap between higher and lower vertebrates.

Carnosine is an endogenous dipeptide abundant in the central nervous system, where by acting as intracellular pH buffering molecule, Zn/Cu ion chelator, antioxidant and anti-crosslinking agent, it exerts a well-recognized multi-protective homeostatic function for neuronal and non-neuronal cells. Carnosine seems to counteract proteotoxicity and protein accumulation in neurodegenerative conditions, such as Alzheimer’s Disease (AD). However, its direct impact on the dynamics of AD-related fibril formation remains uninvestigated. We considered the effects of carnosine on the formation of fibrils/aggregates of the amyloidogenic peptide fragment Aβ1-42, a major hallmark of AD injury. Atomic force microscopy and thioflavin T assays showed inhibition of Aβ1-42 fibrillogenesis in vitro and differences in the aggregation state of Aβ1-42 small pre-fibrillar structures (monomers and small oligomers) in the presence of carnosine. in silico molecular docking supported the experimental data, calculating possible conformational carnosine/Aβ1-42 interactions. Overall, our results suggest an effective role of carnosine against Aβ1-42 aggregation.

The initial observation of peptide absorption in fish intestine dates back to 1981, when, in rainbow trout (Oncorhynchus mykiss), the rate of intestinal absorption of the dipeptide glycylglycine (Gly‐Gly) was compared in vivo with the rate of absorption of its component amino acid glycine (Gly). The description of the identification of the underlying mechanisms that allow di‐ and tripeptide transport across the plasma membranes in fish was provided in 1991, when the first evidence of peptide transport activity was reported in brush‐border membrane vesicles of intestinal epithelial cells of Mozambique tilapia (Oreochromis mossambicus) by monitoring uptake of radiolabelled glycyl‐l‐phenylalanine (Gly‐l‐Phe). Since then, the existence of a carrier‐mediated, H+‐dependent transport of di‐ and tripeptides (H+/peptide cotransport) in the brush‐border membrane of fish enterocytes has been confirmed in many teleost species by a variety of biochemical approaches, providing basic kinetics and substrate specificities of the transport activity. In 2003, the first peptide transporter from a teleost fish, i.e. the zebrafish (Danio rerio) PEPtide transporter 1 (PEPT1), was cloned and functionally characterized in the Xenopus laevis oocyte expression system as a low‐affinity/high‐capacity system. PEPT1 is the protein in brush‐border membranes responsible for translocation of intact di‐ and tripeptides released from dietary protein by luminal and membrane‐bound proteases and peptidases. The transporter possesses affinities for the peptide substrates in the 0.1–10 mM range, depending on the structure and physicochemical nature of the substrates. After the molecular and functional characterization of the zebrafish transporter, the interest in PEPT1 in teleost fish has increased and approaches for cloning and functional characterization of PEPT1 orthologues from other fish species, some of them of the highest commercial value, are now underway. In this paper, we provide a brief overview of the transport of di‐ and tripeptides in teleost fish intestine by recalling the bulk of biochemical, biophysical and physiological observations collected in the pre‐cloning era and by recapitulating the more recent molecular and functional data.

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