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Introducing cell-selectivity in antimicrobial peptides (AMPs) without compromising the antimicrobial and anti-endotoxin properties is a crucial step towards the development of new antimicrobial agents. A peptide designed on phenylalanine heptad repeat possesses significant cytotoxicity along with desired antimicrobial and anti-endotoxin properties. Amino acid substitutions at ‘a’ and/or ‘d’ positions of heptad repeats of AMPs could alter their helical structure in mammalian membrane-mimetic environments and cytotoxicity towards mammalian cells. Since proline is a helix breaker, effects of selective proline substitution(s) at ‘a’ and/or ‘d’ positions of a 15-residue peptide designed on phenylalanine heptad repeat (FR-15) were investigated. Proline-substituted FR-15 variants were highly selective toward bacteria and fungi over hRBCs and murine 3T3 cells and also retained their antibacterial activities at high salt, serum and elevated temperatures. These non-cytotoxic variants also inhibited LPS-induced production of pro-inflammatory cytokines/chemokines in human monocytes, THP-1, RAW 264.7 and in BALB/c mice. The two non-cytotoxic variants (FR8P and FR11P) showed potent anti-cancer activity against highly metastatic human breast cancer cell line MDA-MB-231 with IC 50 values less than 10 μM. At sub-IC 50 concentrations, FR8P and FR11P also showed anti-migratory and anti-invasive effects against MDA-MB-231 cells. FR8P and FR11P induced cellular apoptosis by triggering intrinsic apoptotic pathway through depolarization of mitochondrial membrane potential and activation of caspases. Overall the results demonstrated the utilization of selective phenylalanine to proline substitution in a heptad repeat of phenylalanine residues for the design of cell-selective, broad-spectrum AMPs with significant anti-cancer properties. Statement of Significance We have demonstrated a methodology to design cell-selective potent antimicrobial and anti-endotoxin peptides by utilizing phenylalanine zipper as a template and replacement of phenylalanine residue(s) from “a” and/or “d” position(s) with proline residue(s) produced non-cytotoxic AMPs with improved antibacterial properties against the drug-resistant strains of bacteria. The work showed that the ‘a’ and ‘d’ positions of the phenylalanine heptad repeat could be replaced by an appropriate amino acid to control cytotoxicity of the peptide without compromising its potency in antimicrobial and anti-endotoxin properties. The direct bacterial membrane targeting mechanism of proline substituted analogs of parent peptide makes difficult for bacteria to grow resistance against them. The peptides designed could be lead molecules in the area of sepsis as they possess significant anti-LPS activities for in vitro and in vivo . Interestingly since cancer cells and bacterial cell membranes possess the structural resemblances, the cancer cells are also targets for these peptides making them lead molecules in this field. However, unlike in bacteria where the peptides showed membrane permeabilization property to lyse them, the peptides induced apoptosis in MDA-MB-231 breast cancer cells to inhibit their proliferation and growth. The results are significant because it reveals that “a” and “d” positions of a phenylalanine zipper can be utilized as switches to design cell-selective, antimicrobial, anti - endotoxin and anticancer peptides. [ABSTRACT FROM AUTHOR]

Highlights • The optimization of chromatography for effective separation from interferences. • Extraction optimization of peptides by considering the physicochemical properties. • The non-specific binding assessment of the peptides to the plastic surfaces. • A selective and sensitive method was developed and validated in plasma. • Application of the bioanalytical method to pharmacokinetic studies. Abstract S016-1271 (LR8P) is a broad spectrum novel cationic antimicrobial peptide. The objective of the present study was to develop a selective liquid chromatography–tandem mass spectrometry (LC–MS/MS) based bioanalytical method of S016-1271 peptide in mice and human plasma in order to uncover its pharmacokinetic aspects. The chromatographic separation of S016-1271 (FR8P as internal standard) was achieved on a Waters™ X select CSH-C18 column (75 × 3.0 mm, 2.5 μ) using mixture of acetonitrile and triple distilled water (TDW) both containing 0.05% formic acid as mobile phase. A seven minute linear gradient method was designed to separate analytes from ion suppression at a flow rate of 0.3 mL/min. The extraction of analytes from mice and human plasma was performed through solid phase extraction technique using mixed mode weak cation exchange cartridge (Thermo SOLA WCX 10 mg 1CC) with an extraction recovery of analytes about 75%. Mass spectrometric detection of S016-1271 and FR8P was performed with optimized multiple reaction monitoring (MRM) transitions (Q1/Q3) at 658.8 [M+3H] 3+/653.2 [M+3H-NH 3 ] 3+ and 443.4 [M+5H]5+ /434.7 [y 12 -NH 3 ]4+,respectively in positive electrospray ionization (ESI) mode. The linearity in mice and human plasma was established over a concentration range of 7.81–250 ng/mL with regression coefficient (r2 > 0.99). The currently developed method was validated as per US-FDA guidelines and found to be within the acceptable limits. The method was successfully applied to intravenous (IV) pharmacokinetic study in mice wherein the levels were detected upto 24 h. The peptide demonstrated poor distribution characteristics which were demonstrated through volume of distribution at steady state (202.71 ± 47.02 mL/kg less than total body water of mice; 580 mL/kg). The clearance of the peptide predominantly occurred through central compartment (central clearance is 25 fold greater than peripheral clearance). Also, the in vitro pharmacokinetic studies demonstrated the stability of S016-1271 in plasma and high plasma protein binding in mice and humans. [ABSTRACT FROM AUTHOR]

A variety of CuO nanostructures have been synthesized directly on copper foils by room temperature oxidation of copper through wet chemical method. Alkaline condition necessary for the growth process was maintained by the application of NaOH and NH3 precursors. pH of the solution and reaction time were selected as the process variables. Formation of CuO was confirmed by X-ray diffraction pattern analysis. Evolution of morphologies of the phases formed was characterized by field emission scanning electron microscopy, leading to elucidation of the growth mechanism. Different types of CuO nanostructures were observed to be formed at different process parameters, selected for the present study. Field emission characteristics of CuO nanorods and CuO nanoflakes were also investigated. Emission current density of CuO nanorods and CuO nanoflakes were determined to be 0.90 mA/cm2 and 0.48 mA/cm2, respectively. Huge difference in the emission current density indicates that field emission properties of CuO nanostructures are strongly affected by their morphology. [ABSTRACT FROM AUTHOR]

Dietary fibers are a major source of short-chain fatty acids (SCFAs) in the body, and the fermentation products of SCFAs induced by intestinal microbiota affect energy metabolism. Apart from serving as an energy source in the intestines, SCFAs also inhibit autophagy, nucleotide-binding oligomerization domain-containing protein, LRR, and pyrin domain-containing protein 3 inflammasome. SCFAs provide numerous therapeutic benefits through their influence on cognitive functioning and neurodegenerative diseases (NDD) pathophysiology. Additionally, NDDs are associated with abnormalities in the gut microbiota, including an increased load of pathogens and opportunistic microbes. SCFAs maintain the healthy mitochondrial function and stimulate the maturation of microglia, which consequently suppresses the progression of NDD and cognitive decline by regulating inflammation and oxidative stress. Basically, SCFAs function as cofactors for the host's mitochondrial enzymes and are being studied for their ability to reverse the alteration in the gut microbiota seen in many NDDs and cardiac diseases. In the present review, the focus is on the detrimental and beneficial roles of SCFAs in NDD, emphasizing the effects of SCFA on following phenomenon: (1) alteration in gut microbiota profile associated with NDD, (2) the molecular mechanism of metabolic regulation by SCFA's, and its co-relation with NDD, (3) use of mitochondrial antioxidants as a strategy for maintaining microbiota diversity in the gut, and (4) the future direction of metabolism and neurodegeneration in the gut–brain axis. In addition, the interplay between gut microbiota, SCFAs, epithelial barrier, and neuroimmune signaling in neurodegeneration has been reviewed. The current systematic review and meta-analysis indicate that dietary fermentation of nondigestible fibers leads to the production of short chain fatty acids (SCFAs) by gut microbiota. Moreover, they directly act on the central nervous system and inhibit the progression of different neurodegenerative diseases such as Parkinson disease, Alzheimer disease, and multiple sclerosis. SCFAs act by triggering cellular antioxidants and inhibiting inflammation. The properties of SCFAs depend on the G protein–coupled receptors, histone deacetylases and peroxisome proliferator-activated receptor gamma, and regulatory T-cell activation. [Display omitted] [ABSTRACT FROM AUTHOR]

Highlights: [•] Photoexcited 97/78 generated photoproduct [i.e. (m/z=376.7)] identified by LC–MS/MS. [•] Singlet oxygen generation via type-II dependent mechanism. [•] 2′-dGuO and Comet results illustrated the photogenotoxic potential. [•] Cell cycle arrest in G2/M, lysosomal disruption and MMP loss. [•] Upregulation of p21, Bax and caspase-3 and apoptosis. [ABSTRACT FROM AUTHOR]