Attenuation of COX-2 enzyme by modulating H2O2-mediated NF-κB signaling pathway by monoamine oxidase inhibitor (MAOI): a further study on the reprofiling of MAOI in acute inflammation.

OBJECTIVE: This study aims to investigate the anti-inflammatory mechanism of monoamine oxidase inhibitor (MAOI) in carrageenan (CARR) induced inflammation models to reprofile their use. We also aimed to explore the role of monoamine oxidase (MAO)-mediated H2O2-NF-κB-COX-2 pathway in acute inflammation. METHODS: In vitro anti-inflammatory activity and hydrogen peroxide (H2O2) scavenging activity were performed according to the established procedure. Inflammation was induced using CARR in BALB/c mice at the foot paw and peritoneal cavity. Hourly measurement of paw swelling was performed. The level of nitric oxide (NO), myeloperoxidase (MPO), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2) and nuclear factor κB (NF-κB) was determined using enzyme-linked immunosorbent assay (ELISA). Peritoneal fluid was collected to investigate total count, differential count of leukocytes, and capillary permeability. RESULTS: In vitro anti-inflammatory evaluations revealed the potential role of MAOI to inhibit heat-induced protein denaturation and human red cell membrane destabilization. H2O2 inhibition activity of MAOI also proved their powerful role as an H2O2 scavenger. Treatment with MAOI in CARR-induced mice significantly reduced paw edema, leukocyte extravasation, and total and differential leukocyte count. The result of ELISA showed MAOI effectively reduce the level of COX-2, PGE2 and NF-κB in inflamed tissue. CONCLUSIONS: In short, this study demonstrates that inhibition of H2O2 by MAOI alleviates CARR-induced paw edema possibly by inhibiting the H2O2-mediated NF-κB-COX-2 pathway. The present investigation identifies MAOI might reprofile for the treatment of acute inflammation also, the MAO enzyme may use as a novel therapeutic target to design and develop new class of anti-inflammatory agents.

Repurposing monoamine oxidase inhibitors (MAOI) for the treatment of rheumatoid arthritis possibly through modulating reactive oxidative stress mediated inflammatory cytokines.

Monoamine oxidase inhibitors (MAOI) are presently used to treat depression, parkinsonian, and other psychiatric disorders. The present study was aimed to repurpose the use of MOAI in Rheumatoid Arthritis (RA). The animal model of RA was developed using collagen type II (CII) in Freund's complete adjuvant (FCA) followed by lipopolysaccharide (LPS) and a booster dose of CII in FCA. The effect of MAOI, Selegiline was evaluated whereas the indicators like paw thickness, arthritic score, and the splenic index were measured and compared with the standard drug Methotrexate. Further to explore the molecular mechanism, the expression of serum inflammatory cytokines (IL-6 and TNF-α), radiographical and histopathological study of hind paw were also checked and analyzed. Treatment with MAOI, Selegiline not only reduced the paw thickness, arthritic score, and the splenic index, but also greatly improved the inflammatory biochemical and hematologic parameters and improved the arthritis score. The serum level of IL-6 and TNF-α are considerably decreased dose dependently, however, the notable significant effect (**p < 0.01) observed at concentration of 30 mg/kg b.w. when the RA animals treated by Selegiline. Collectively, Selegiline improved the progression of RA possibly via decreased catecholamine breakdown at synovial fluid resulting decrease hydrogen peroxide (H2O2) generation and inhibition of pro-inflammatory cytokines in situ. Thus, the finding support and indicate the repurposing of MAOI for the treatment of RA meriting further studies on synovial monoamine oxidase as a new therapeutic target to design a new drug for RA.

Structure-based discovery of (S)-2-amino-6-(4-fluorobenzyl)-5,6,11,11a-tetrahydro-1H-imidazo[1',5':1,6]pyrido[3,4-b]indole-1,3(2H)-dione as low nanomolar, orally bioavailable autotaxin inhibitor.

Inhibition of extracellular secreted enzyme autotaxin (ATX) represents an attractive strategy for the development of new therapeutics to treat various diseases and a few inhibitors entered in clinical trials. We herein describe structure-based design, synthesis, and biological investigations revealing a potent and orally bioavailable ATX inhibitor 1. During the molecular docking and scoring studies within the ATX enzyme (PDB-ID: 4ZGA), the S-enantiomer (Gscore = -13.168 kcal/mol) of the bound ligand PAT-494 scored better than its R-enantiomer (Gscore = -9.562 kcal/mol) which corroborated with the reported observation and analysis of the results suggested the scope of manipulation of the hydantoin substructure in PAT-494. Accordingly, the docking-based screening of a focused library of 10 compounds resulted in compound 1 as a better candidate for pharmacological studies. Compound 1 was synthesized from L-tryptophan and evaluated against ATX enzymatic activities with an IC50 of 7.6 and 24.6 nM in biochemical and functional assays, respectively. Further, ADME-PK studies divulged compound 1 as non-cytotoxic (19.02% cell growth inhibition at 20 µM in human embryonic kidney cells), metabolically stable against human liver microsomes (CLint  = 15.6 µl/min/mg; T1/2  = 113.2 min) with solubility of 4.82 µM and orally bioavailable, demonstrating its potential to be used for in vivo experiments.

Role of Glycogen Synthase Kinase-3 in the Etiology of Type 2 Diabetes Mellitus: A Review.

The risk of type 2 diabetes mellitus (T2DM) is increasing abundantly due to lifestyle-related obesity and associated cardiovascular problems. Presently, Glycogen synthase kinase-3 (GSK-3) has gained considerable attention from biomedical scientists to treat diabetes. Phosphorylation of GSK-3 permits a number of cellular activities like regulation of cell signaling, cellular metabolism, cell proliferation and cellular transport. Inhibiting GSK-3 activity by pharmacological intervention has become an important strategy for the management of T2DM. This review focuses on the schematic representation of fundamental GSK-3 enzymology and encompasses the GSK-3 inhibitors as a future therapeutic lead target for the management of T2DM that may significantly regulate insulin sensitivity to insulin receptor, glycogen synthesis and glucose metabolism. The various signaling mechanisms of inhibiting the GSK-3 by describing insulin signaling through Insulin Receptor Substrate (IRS-1), Phosphatidylinositol-3 Kinase (PI3K) and Protein Kinase B (PKB/ AKT) pathways that may hopefully facilitate the pharmacologist to design for antidiabetic drug evaluation model in near future have also been highlighted.

Zebra Fish in Toxicology Research: Streptavidin Conjugated Peroxidase Assay in the Development Phase of Zebrafish Embryos to Study Liver Toxicities.

BACKGROUND: Zebrafish have similar hepatic anatomy and cellular architecture just like mammals. Therefore, a number of investigators are using zebrafish to study liver pathologies. However, the evaluation model specific to liver toxicities in zebra fish was not clearly stated earlier. AIMS AND OBJECTIVES: The present study was designed to develop a model of embryonic liver toxicity using dexamethasone (DEXA, 0-20 µM) as a standard hepatotoxic agent. METHODS: such toxicities are easily measured by streptavidin-conjugated peroxidase assay after 48- hour post-fertilization (hpf) of DEXA treatment. RESULTS: In addition to morphological toxicities at different hpf, DEXA showed significant (*p< 0.05 &**p<0.01) reduction of peroxidase-chromogenic dye reaction in the assay as compared to DEXA untreated embryos at 10 & 20 µM concentration that concluded the hepatocellular toxicity of dexamethasone. CONCLUSION: Hopefully, the developed model for hepatotoxicity evaluation will be a promising model for the evaluation of new drugs or chemicals as an easy vertebrate model before the commencement of another animal model.

Frequency Dependent Charge Transport and Spin State Switching Characteristics of Fe(phen)2(NCS)2 in Polymer.

We report on the bistability in spin states of spin crossover (SCO) compound Fe(phen)2(NCS)2 in polymer (polypyrrole) by frequency (1-100 kHz) and temperature dependent (305-457 K) electrical conductivity measurements. The structure and growth of SCO compounds in conducting polymer are obtained by scanning electron microscopy, X-ray diffraction and optical absorption measurements. The thermal dependence of ac conductivity σ(ω) shows the clear formation of a hysteresis loop in its cooling and heating cycle due to the difference in conductivity in high spin and low spin state. The size, shape and width of the hysteresis loops are found to be critically dependent on the applied frequency and/or the ratio between SCO and polymer. The ac conductivity is found to exhibit a dispersive behavior following Jonscher's law: σ(ω) ∝ ωn below a critical frequency ωc, above which it is found to monotonically decrease with increasing frequency. The thermal dependence of the exponent n and ωc is also explored. The charge transport phenomena are explained in the framework of hopping of charge carriers. The data reveals that addition of polymer can play an important role to tune the conductivity of SCO compounds and its spin state dependence characteristics which may be quite helpful for fabricating future spin-based devices. Temperature dependent magnetic susceptibility measurement also confirms the spin transition behavior of the SCO/ppy composite samples. These SCO/ppy composite samples can be taken as the reliable nanomaterials fabricated with the concept of future spin based nanoarchitectonics.

Probing Spin-State Switching in Fe(phen)2(NCS)2 Thin Film Nanocrystals on Different Substrates by Electrical Conductivity Measurements.

Spin-state switching mechanism is investigated by measuring the temperature of the electrical conductivity of spin crossover (SCO) material Fe(phen)2(NCS)2 thin films grown on glass, quartz and silicon substrates. The morphology characterized by scanning electron microscopy, clearly reveals the growth of thin films of thickness ~300 nm comprising of nanocrystals, size and distribution of which is dependent on the nature of substrates. The film on quartz is found to have the most uniform growth of nanocrystals of size ~22 nm with a homogeneous distribution. All the films retain the orthorhombic crystal structure as that of bulk with slight distortions in lattice plausibly arising out of the strain. Spin state switching between LS and HS is clearly revealed by the hysteresis loop observed in the temperature dependence of the electrical conductivity in its heating and cooling cycle. The critical temperature of transition between HS and LS states is found to be 162 K, 193 K and 217 K for film on glass, quartz and Si respectively. Film on quartz is found to exhibit a wide hysteresis loop of width ~60 K while that of on silicon exhibits higher transition temperature with narrow hysteresis loop ~14 K. The results are found to be quite inspiring to tune the SCO characteristics to develop molecular switch and memory devices close to room temperature.

Oxidative stress in inflammatory cells of patient with rheumatoid arthritis: clinical efficacy of dietary antioxidants.

Rheumatoid arthritis (RA) is an autoimmune disease responsible for significant human morbidity in modern life. However, oxidative stress is one of the key markers for determining pathophysiology of patients with RA. The interaction between cellular immune system and body's endogenous and/or exogenous antigens produce reactive oxygen species (ROS) and reactive nitrogen species (RNS) in autoimmune disease like RA. ROS and RNS include highly toxic superoxide (O2-) and peroxynitrite (ONOO-) radicals, which activate the signaling cascades of inflammatory cells to synthesize pro-inflammatory cytokines and chemokines. Previous studies reported that Th1 cytokines could promote the development of autoimmune disorders like RA, whereas the Th2 cytokines may attenuate the same diseases. An increased awareness of the relationship between food and health led to a tremendous increase of antioxidant research in the last decade. Evaluation of the efficacy of dietary antioxidants is also becoming highly acceptable in RA research. A number of dietary phytomolecules are already established as having antioxidant activity in isolated synovial cellular infiltrate or peripheral blood neutrophils and lymphocytes. This review aims to highlight the oxidative stress in inflammatory cells of patients with RA and to summarize the clinical relevance of dietary antioxidants as a first step in assessing beneficial effect, safety and dose safety ratio in patients with RA.

Antimicrobial properties of Kalanchoe blossfeldiana: a focus on drug resistance with particular reference to quorum sensing-mediated bacterial biofilm formation.

OBJECTIVES: This study attempts to investigate the antimicrobial properties of Kalanchoe blossfeldiana with a particular reference to quorum sensing (QS)-mediated biofilm formation. METHODS: The methanol extract of K. blossfeldiana leaves (MEKB) was evaluated for antimicrobial properties including QS-controlled production of biofilm (including virulence factor, motility and lactone formation) in Pseudomonas aeruginosa. Methanol extract of K. blossfeldiana was also evaluated for anti-cytokine (tumour necrosis factor-alpha, interleukin-6 and interleukin-1 beta) properties in peripheral blood mononuclear cells (PBMC). KEY FINDINGS: Methanol extract of K. blossfeldiana exhibited antimicrobial effect on clinical isolates, as well as standard reference strains. Pseudomonas aeruginosa exposed to MEKB (subminimum inhibitory concentration (MIC)) displayed reduced biofilm formation, whereas supra-MIC produced destruction of preformed biofilms. Methanol extract of K. blossfeldiana reduced the secretion of virulence factors (protease and pyoverdin) along with generation of acyl homoserine lactone (AHL). Confocal laser scanning microscopy images indicate reduction of biofilm thickness. The extract also reduced cytokine formation in lipopolysaccharide-stimulated PBMC. CONCLUSIONS: Kalanchoe blossfeldiana was found to interfere with AHL production, which in turn may be responsible for downregulating QS-mediated production of biofilm and virulence. This first report on the antibiofilm and anticytokine properties of this plant may open up new vistas for future exploration of this plant for combating biofilm-related resistant infections.