Concentration- and Solvent-Induced Chiral Tuning by Manipulating Non-Proteinogenic Amino Acids in Glycoconjugate Supra-Scaffolds: Interaction with Protein, and Streptomycin Delivery.

We showed solvent- and concentration-triggered chiral tuning of the fibrous assemblies of two novel glycoconjugates Z-P(Gly)-Glu and Z-F(4-N)-Glu made by chemical attachment of Cbz-protected [short as Z)] non-proteinogenic amino acids L-phenylglycine [short as P(Gly)] and 4-Nitro-L-phenylalanine [short as F(4-N)] with D-glucosamine [short as Glu]. Both biomimetic gelators can form self-healing and shape-persistent gels with a very low critical gelator concentration in water as well as in various organic solvents, indicating they are ambidextrous supergelators. Detailed spectroscopic studies suggested ß-sheet secondary structure formation during anisotropic self-aggregation of the gelators which resulted in the formation of hierarchical left-handed helical fibers in acetone with an interlayer spacing of 2.4 nm. After the physical characterization of the gels, serum protein interaction with the gelators was assessed, indicating they may be ideal for biomedical applications. Further, both gelators are benign, non-immunogenic, non-allergenic, and non-toxic in nature, which was confirmed by performing the blood parameters and liver function tests on Wister rats. Streptomycin-loaded hydrogels showed efficacious antibacterial activity in vitro and in vivo as well. Finally, cell attachment and biocompatibility of the hydrogels were demonstrated which opens a newer avenue for promising biomedical and therapeutic applications.

An improved method for experimental induction of ulcerative colitis in Sprague Dawley rats.

Ulcerative colitis (UC) is a chronic inflammatory manifestation of the human colon that is linked with colorectal cancer. Development of an appropriate animal model is crucial to study the immunopathophysiology of UC wherein chemical induction is the most popular method of choice. However, unavailability of an optimum experimental model limits the success of this method. The present study aims to establish an optimized model for acetic acid-induced colitis in Sprague Dawley rats. Response Surface Methodology (RSM) with a six-factors Box-Behnken design was employed to generate an improved method of inducing UC in rat, predicting the case statistics, apposite investigation of quadratic response surfaces, and construction of a second-order polynomial equation. UC was diagnosed through three responses viz. weight loss, severity of diarrhea, and appearance of blood in the stool. Analysis of variance alongside RSM jointly revealed that induction of UC can be achieved with highest probability using the combination of parameters that includes 120 gm body weight, 1.5 ml of 4% acetic-acid v/v in distilled water with a single dose of treatment for 24 h including a pre-induction of 5 mins. This optimized UC-induction model was validated in-vivo through disease scoring index and hematological assessments with satisfactory level of desirability. •An improved experimental method for inducing ulcerative colitis (UC) in Sprague Dawley rats has been developed.•Box-Behnken Design-fitted Response Surface Methodology (RSM) was implicated in optimizing the experimental parameters for generating UC.•This statistically optimized and experimentally validated method resembles the recipe for the generation of UC in animal model with the highest possible desirability.

Cell Surface Fibroblast Activation Protein-2 (Fap2) of Fusobacterium nucleatum as a Vaccine Candidate for Therapeutic Intervention of Human Colorectal Cancer: An Immunoinformatics Approach.

Colorectal cancer (CRC) is one of the most common cancers and is the second-highest in cancer-related deaths worldwide. The changes in gut homeostasis and microbial dysbiosis lead to the initiation of the tumorigenesis process. Several pathogenic gram-negative bacteria including Fusobacterium nucleatum are the principal contributors to the induction and pathogenesis of CRC. Thus, inhibiting the growth and survival of these pathogens can be a useful intervention strategy. Fibroblast activation protein-2 (Fap2) is an essential membrane protein of F. nucleatum that promotes the adherence of the bacterium to the colon cells, recruitment of immune cells, and induction of tumorigenesis. The present study depicts the design of an in silico vaccine candidate comprising the B-cell and T-cell epitopes of Fap2 for improving cell-mediated and humoral immune responses against CRC. Notably, this vaccine participates in significant protein-protein interactions with human Toll-like receptors, especially with TLR6 reveals, which is most likely to be correlated with its efficacy in eliciting potential immune responses. The immunogenic trait of the designed vaccine was verified by immune simulation approach. The cDNA of the vaccine construct was cloned in silico within the expression vector pET30ax for protein expression. Collectively, the proposed vaccine construct may serve as a promising therapeutic in intervening F. nucleatum-induced human CRC.