Insights from in silico exploration of major curcumin analogs targeting human dipeptidyl peptidase IV.

The goal of this work is to use a variety of in-silico techniques to identify anti-diabetic agents against DPP-IV enzyme from five main curcumin analogues. To produce the successful molecules, five main curcumin analogues were docked into the active site of DPP-IV enzyme. In comparison to the control molecule (Saxagliptin, -6.9 kcal/mol), all the compounds have the highest binding affinity (-7.6 to -7.7 kcal/mol) for the DPP-IV enzyme. These compounds underwent further testing for studies on drug-likeness, pharmacokinetics, and acute toxicity to see the efficacy and safety of compounds. To assess the stability of the docking complex and the binding posture identified during the docking experiment, our study got THC as the lead compound, which was then exposed to 200 ns of molecular dynamic simulation and PCA analysis. Additionally, DFT calculations were conducted to determine the thermodynamic, molecular orbital, and electrostatic potential characteristics of lead compound. Overall, the lead chemical has shown strong drug-like properties, is non-toxic, and has a sizable affinity for the DPP-IV enzyme.Communicated by Ramaswamy H. Sarma.

Diabetic wound healing of aloe vera major phytoconstituents through TGF-ß1 suppression via in-silico docking, molecular dynamic simulation and pharmacokinetic studies.

To restore the integrity of the skin and subcutaneous tissue, the wound healing process involves a complex series of well-orchestrated biochemical and cellular events. Due to the existence of various active components, accessibility and few side effects, some plant extracts and their phytoconstituents are recognised as viable options for wound healing agents. To find possible inhibitors of diabetic wound healing, four main constituents of aloe vera were identified from the literature. TGF-ß1 and the compounds were studied using molecular docking to see how they interacted with the active site of target protein (PDB ID: 6B8Y). The pharmacokinetics investigation of the aloe emodin with the highest dock score complied with all the Lipinski's rule of five and pharmacokinetics criteria. Conformational change in the docked complex of Aloe emodin was investigated with the Amber simulation software, via a molecular dynamic (MD) simulation. The MD simulations of aloe emodin bound to TGF-ß1 showed the significant structural rotations and twists occurring from 0 to 200 ns. The estimate of the aloe emodin-TGF-ß1 complex's binding free energy has also been done using MM-PBSA/GBSA techniques. Additionally, aloe emodin has a wide range of enzymatic activities since their probability active (Pa) values is >0.700. 'Aloe emodin', an active extract of aloe vera, has been identified as the key chemical in the current investigation that can inhibit diabetic wound healing. Both in-vitro and in-vivo experiments will be used in a wet lab to confirm the current computational findings.Communicated by Ramaswamy H. Sarma.

Computational evaluation of quinones of Nigella sativa L. as potential inhibitor of dengue virus NS5 methyltransferase.

Aedes aegypti is the primary vector for the transmission of the dengue virus, which causes dengue fever, dengue hemorrhagic illness and dengue shock syndrome. There is now no antiviral medication available to treat DENV, which kills thousands of people each year and infects millions of individuals. A possible target for the creation of fresh and efficient dengue treatments is the DENV-3 NS5 MTase. So, Nigella sativa quinones were examined using in silico methods to find natural anti-DENV compounds. The in silico docking was conducted utilising the Discovery Studio software on the quinones of N. sativa and the active site of the target protein DENV-3 NS5 MTase. In addition, the druggability and pharmacokinetics of the lead compound were assessed. Dithymoquinone was comparable to the reference compound in terms of its ability to bind to the active site of target protein. Dithymoquinone met the requirements for drug likeness and Lipinski's principles, as demonstrated by the ADMET analysis and drug likeness results. The current study indicated that the dithymoquinone from N. sativa had anti-DENV activity, suggesting further drug development and dengue treatment optimisation.Communicated by Ramaswamy H. Sarma.

Usnic acid as potential inhibitors of BCL2 and P13K protein through network pharmacology-based analysis, molecular docking and molecular dynamic simulation.

Usnic acid (UA) lately piqued the interest of researchers for its extraordinary biological characteristics, including anticancer activity. Here, the mechanism was clarified through network pharmacology,molecular docking and molecular dynamic simulation. Sixteen proteins were selected through network pharmacology study as they are probable to interact with UA. Out of these proteins, 13 were filtered from PPI network analysis based on their significance of interactions (p < 0.05). KEGG pathway analysis has also aided us in determining the three most significant protein targets for UA, which are BCL2, PI3KCA and PI3KCG. Therefore molecular docking and molecular dynamic (MD) simulations throughout 100 ns were performed for usnic acid onto the three proteins mentioned. However, UA's docking score in all proteins is lower than their co-crystalised ligand, especially for BCL2 (-36.5158 kcal/mol) and PI3KCA (-44.5995 kcal/mol) proteins. The only exception is PI3KCG which has comparable results with the co-crystallised ligand with (-41.9351 kcal/mol). Furthermore, MD simulation has also revealed that usnic acid does not stay fit in the protein throughout the simulation trajectory for PI3KCA protein evident from RMSF and RMSD plots. Nevertheless, it still poses good ability in inhibiting BCL2 and PI3KCG protein in MD simulation. In the end, usnic acid has exhibited good potential in the inhibition of PI3KCG proteins, rather than the other proteins mentioned. Thus further study on structural modification of usnic acid could enhance the ability of usnic acid in the inhibition of PI3KCG as anti-colorectal and anti-small cell lung cancer drug candidate.Communicated by Ramaswamy H. Sarma.

Comparative Study of Onion (Allium cepa) and Leek (Allium ampeloprasum): Identification of Organosulphur Compounds by UPLC-QTOF/MS and Anticancer Effect on MCF-7 Cells.

Onion (Allium cepa) and leek (Allium ampeloprasum var. porrum) are common herbs and vegetables found in our daily life. It belongs to the genus Allium, which is usually known for their high antioxidant and anticancer properties. Medical researchers highly recommend the exploitation of herbs and plants as alternative ways in the treatment of cancer. This research was designed to study the anticancer effects of onion and leek extracts on MCF-7 human breast cancer cell. Crude extracts of ethanol, methanol, and water of onion and leek were obtained by maceration. MCF-7 cells were cultured in complete media at 37 °C and subjected to different treatments that involved varying concentrations (10, 50, and 100 µg/mL) of onion and leek extracts for 24, 48, and 72 h of incubation. The percentage of cell viability and the concentration of extracts on MCF-7 cells were determined using MTT assay. The water leek extract proved to be the most effective extract at 50 µg/mL, whereby it showed a significant inhibition ability due to the presence of entadamide A-ß-D-glucopyranoside as identified by ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF/MS). Further studies about the mechanism of both extracts in causing cell death and the determination of the presence of other bioactive compounds in the extracts are needed.

Cellulose Supported Pd(II) Complex Catalyzed Carbon-Carbon Bonds Formation.

Corn-cobs are an agro-industrial waste and composed of cellulose mostly. In this study cellulose was isolated from the waste corn-cobs and modified to polymeric hydroxamic acid palladium complex 1 and characterized by using a variety of spectroscopic methods such as field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and inductively coupled plasma atomic emission spectroscopy (ICP-AES). The complex 1 exhibited high catalytic activity towards Suzuki and Heck coupling reactions of activated and deactivated aryl halides to give the respective coupling products with high yield. Moreover, the complex 1 was recovered and recycled five times with no considerable loss of catalytic overall performance.

UPLC-QTOF/MS-based phenolic profiling of Melastomaceae, their antioxidant activity and cytotoxic effects against human breast cancer cell MDA-MB-231.

Eleven compounds were identified during profiling of polyphenols by UPLC-QTOF/MS. In abundance was quercetin-3-O-α-l-arabinofuranoside in M. malabathricum ethanolic leaves extract while 6-hydroxykaempferol-3-O-glucoside was present in the leaves extract of M. decenfidum (its rare variety). TPC and TFC were significantly higher in M. decemfidum extract than M. malabathricum extract. During DPPH, FRAF and ß-carotene bleaching assays, M. decemfidum extract exhibited greater antioxidant activity compared to M. malabathricum extract. Effect of M. malabathricum and M. decemfidum extracts on viability of MDA-MB-231 cell at concentrations 6.25-100 µg/mL were evaluated for 24, 48 and 72 h. After 48 and 72 h treatment, M. malabathricum and M. decemfidum leaves extracts exhibited significant activity in inhibiting MDA-MB-231 cancer cell line with M. malabathricum extract being more cytotoxic. M. malabathricum and M. imbricatum serves as potential daily dietary source of natural phenolics and to improve chemotherapeutic effectiveness.

Characterization of anti-Tn-antigen MLS128 binding proteins involved in inhibiting the growth of human colorectal cancer cells.

MLS128 monoclonal antibody, which binds an epitope consisting of two or three consecutive Tn-antigens, inhibits colon cancer cell growth by binding to a 110 kDa glycoprotein (GP). Previous studies suggested a possible association of insulin-like growth factor-I receptor (IGF-IR) signaling in the inhibition of colon cancer cell growth by MLS128 (Morita et al. Biosci Trends. 3, 32-37, 2009; Zamri et al. ibid. 6, 303-312, 2012). The current study thus investigated the nature of 110 kDa GP and its possible association with IGF-IR. MLS128 treatment for 3 days caused down-regulation of IGF-IR and disappearance of 110 kDa GP in HT29 colon cancer cells. Immunoprecipitation/immunoblotting experiments did not reveal a direct association between the two molecules in HT29 cells. In LS180 and HT29 cells, however, 110 kDa GP and IGF-IR were found in microdomains. Treatment of these cells with MLS128 for 3 days caused a reduction in the IGF-IR and 110 kDa GP associated with microdomains. Two-dimensional gel electrophoresis/MLS128 immunoblotting of HT29 and LS180 cell lysates and immunoprecipitates revealed three spots, from which tryptic peptides were recovered for protein sequencing. Identification of 110 kDa GP was unsuccessful due to its heterogeneity and resistance to tryptic digestion. During this study, however, limited proteolysis of 110 kDa GP was observed in the microdomain-associated 110 kDa GP from HT29 and LS180 cells, suggesting that protease-susceptible sites or domains exist in the middle of 110 kDa GP. This information on limited proteolysis may provide a clue to identifying 110 kDa GP.

A novel combined adjuvant for nasal delivery elicits mucosal immunity to influenza in aging.

Since a combination of flt3 ligand plasmid (pFL) and CpG-oligodeoxynucleotides (ODN)(3) as a dendritic cell (DC)-targeting double mucosal adjuvant elicited ovalbumin-specific secretory IgA (S-IgA) antibody (Ab) responses, we examined whether this double adjuvant could induce influenza-specific protective immunity in aged mice. A double adjuvant plus A/Puerto Rico/8/34 (PR8) hemagglutinin (HA) induced increased numbers of CD11b(+) CD11c(+) DCs and both CD4(+) Th1- and Th2-type responses in the nasopharyngeal-associated lymphoreticular tissue, nasal passages and cervical lymph nodes. Further, increased levels of PR8 HA-specific S-IgA Ab responses were detected in the upper respiratory tact (URT) of aged and young adult mice given nasal PR8 HA with this double adjuvant. Thus, when mice were challenged with PR8 virus via the nasal route, both aged and young adult mice given nasal vaccine exhibited complete protection. Further, IgA-deficient mice nasally immunized with a double adjuvant influenza vaccine failed to provide protection against PR8 challenge. These results indicate that a nasal double adjuvant successfully induces PR8 HA-specific IgA Ab responses in both young adult and aged mice, which are essential for the prevention of influenza infection in the murine URT.

Characterization of three different single chain antibodies recognizing non-reducing terminal mannose residues expressed in Escherichia coli by an inducible T7 expression system.

We previously isolated phage antibodies from a phage library displaying human single chain antibodies (scFvs) by screening with a mannotriose (Man3)-bearing lipid. Of four independent scFv genes originally characterized, 5A3 gene products were purified as fusion proteins such as a scFv-human IgG1 Fc form, but stable clones secreting 1A4 and 1G4 scFv-Fc proteins had never been established. Thus, bacterial expression systems were used to purify 1A4 and 1G4 scFv gene products as soluble forms. Purification of 1A4 and 1G4 scFv proteins from inclusion bodies was also carried out together with purification of 5A3 scFv protein in order to compare their Man3-binding abilities. The present studies demonstrated that 1A4 and 1G4 scFv proteins have a higher affinity for Man3 than 5A3 scFv protein, which may determine whether scFv-Fc proteins expressed in mammalian cells are retained in the ER or secreted. Furthermore, the inhibitory effects of anti-Man3 1G4 scFv and anti-Tn antigen scFv proteins on MCF-7 cell growth were evaluated. Despite the fact that no obvious difference was detected in cell growth, microscopic observations revealed inhibition of foci formation in cells grown in the presence of the anti-carbohydrate scFv proteins. This finding provides a basis for the development of cancer therapeutics.