Decreased levels of serum soluble complement receptor-II (CR2/CD21) in patients with rheumatoid arthritis

Objective. The soluble cluster of differentiation 21 (sCD21) represents the extracellular portion of the CD21 glycoprotein and is released by shedding from cell surfaces into plasma. Soluble CD21 binds complement fragments and activates monocytes through binding to membrane CD23. Elevated levels of sCD21 are found during Epstein-Barr virus EBV infections, B-cell lymphoma and other lymphoblastoid tumours. The present study was undertaken to investigate levels of sCD21 in rheumatoid arthritis. Methods. A specific enzyme-linked immunoassay was developed using sCD21, biochemically purified to homogeneity from human plasma as a standard for the determination of sCD21 concentration in patient sera. Peripheral blood B and T lymphocytes were isolated from healthy donors and rheumatoid arthritis patients and cultured, and supernatants were analysed for CD21 shedding. Results. The normal values of serum sCD21 in healthy individuals between 20 and 40 yr of age ranged from 100 to 477 ng/ml (median 292 ng/ml), decreasing with age but not differing with gender. In rheumatoid arthritis patients, sCD21 levels ranged from 50 to 300 ng/ml (median 182 ng/ml), did not differ with age and were independent of rheumatoid factor. Conclusions. In contrast to healthy donors, patients with rheumatoid arthritis have significantly lower sCD21 levels (P < 0.0001), independently of the age of the patients. Sorted B cells from rheumatoid arthritis patients released amounts of CD21 comparable with those of normal controls. Possible causes and consequences of the findings are discusse

NOVEL IN SILICO APPROACH OF ANTICANCER ACTIVITY BY INHIBITING HEMOPEXIN PROTEINS WITH INDIGOFERA ASPALATHOIDESPLANT CONSTITUENTS AT ACTIVE SITE

Objective: The aim of the study was to investigate the anti-cancer activity using the phytoconstituents of Indigofera aspalathoides.Methods: The plant extract has been largely used as cell proliferation inhibitors. In this study, specific phytoconstituent has been targeted towardsmatrix metalloproteinases (MMPs).Results: MMPs are group of proteinases that are associated with cell invasion inhibition and also inhibit proliferation. The C-terminal domain ofMMPs mimics the serum protein hemopexin (HPX). According to various literatures, a reason for the failure of MMP as anti-cancer agent is thepresence of this HPX binding at the active site.Conclusion: A novel approach was carried to inhibit this binding by Carotal, (-)-Spathulenol, Tau.-Cadinol proteins from the plant I. aspalathoides. Keywords: Hemopexin, Matrix metalloproteinase, Indigofera aspalathoides, Molecular docking, Carotal, (-)-Spathulenol, Tau.-Cadinol

Associated technologies ensures complete loop mediated isothermal amplification platform for pathogen diagnosis

Loop Mediated Isothermal Amplification (LAMP) assay could be a useful adjunct diagnostic assay along with the conventional methods that would preclude the requirement of continuous maintenance of pure cultures. Moreover, LAMP assay is simple, rapid, specific and sensitive for the detection of pathogens. Having developed and validated LAMP method for the detection of an isolated pathogen, Escherichia coli O157:H7, an attempt was made to progress the LAMP platform to realistic point of care for resource-poor endemic areas. Reaction time of the LAMP method was only 1 h and also, the amplification products of O157, which had the corresponding target genes, turned green by visual inspection when added with Calcein/Sybr green. However, sample preparation and lyophilized master mix preparation before LAMP assay as well as developing a closed detection system for detection of LAMP amplified products remained a quest. Hence, the current study was conducted to develop a lyophilized LAMP master mix for easy platform to take LAMP to realistic point of care and Dot-Elisa based ‘lateral flow dipstick’ that could ease the detection of LAMP products in a closed environment. Sample preparation is another associated technology that is yet-to-be developed.Keywords: Loop mediated isothermal amplification assay, polymerase chain reaction, lyophilized master mix, and closed amplification system.African Journal of Biotechnology Vol. 12(42), pp. 6049-605

ANALYSIS OF MOLECULAR DOCKING EFFICIENCY OF CLEISTANTHIN-A, AS AN ALTERNATIVE FOR NICOTINE ADDICTION

Objective: The present research was aimed to understand the molecular docking efficiency of a plant-derived compound cleistanthin-A and a common ingredient in tobacco consumption nicotine with nicotinic acetylcholine receptor (nAChR).Methods: The 3-D structure of nAChR was retrieved from the protein data bank (ID 5AFH). Ligand was obtained from the PUBCHEM. The in silico protocol comprised of three steps: high-throughput virtual screening (HTVS), standard preci­sion (SP) and extra precision (XP). The screened molecules were ranked accordingly using glide score. Schrödinger tool was used to perform the docking analysis.Results: The binding efficiency of the nicotine and cleistanthin-A was found to be docked at the cys-cys loop of the receptor. Based upon the glide score and glide energy it can be reported that, nicotine binding can be inhibited by the binding of cleistanthin-A to the nAChR.Conclusion: The docking efficiency of cleistanthin-A was good compared to nicotine towards nAChR. Hence, cleistanthin–A was derived as a better choice as an alternative for nicotine in smoke therapy

Mass Spectrometry-based Sequencing of Venom Peptides (Conotoxins) from Vermivorous Cone Snail, Conus Loroisii: Toxicity of its Natural Venom

Conus loroisii is a marine vermivorous snail found profusely in the southern seas of India. They harbor several toxic peptide components commonly called as ‘conotoxins’. In this study, we have identified and sequenced five conotoxins using proteome based tandem mass spectrometry analysis through Data analysis 4.1 software. Among them, we found Lo959 as contryphan which is previously described. All other conotoxins Lo1702, Lo1410, Lo1385 and Lo1686 belong to M-Superfamily conotoxins and novel to C. loroisii. Lo1410 is completely novel to conotoxin research with 3 disulfides and the amino acid sequence is derived as CCSTNCAVCIPCCP. All the identified M-Superfamily conotoxins are sub categorised to mini M2 superfamily conotoxins. Lo1702 and Lo1686 possess C- terminal amidation which is the key feature in conotoxins. Moreover, we have screened the natural venom for the occurrence of toxicity in the zebrafish model and brine shrimp

Evidence for the Double Excimer State of conjugated polymer in a liquid solution

In this paper, the spectral properties of a conjugated polymer poly [2-methoxy-5-(2-ethylhexyloxy)-1, 4-phenylenevinylene] (MEH-PPV) in benzene have been studied. The results showed that the fluorescence spectra of MEH-PPV under low concentrations had two peaks; the dominant one due to monomer was around 560 nm, and the shoulder one attributed to the excimer was around 600 nm. Under higher concentrations, it was found that there was only one band around 600 nm due to the excimeric state. By increasing the concentrations of MEH-PPV, it could be seen that there was a new band around 640nm. This band is being attributed to the double excimer. Under high power pulsed laser excitation, we observed amplified spontaneous emission (ASE) at 570 nm, 605 nm and 650 nm. These ASE peaks could arise from the monomer, excimer and double excimer states of the macromolecule respectively. To the best of our knowledge this is perhaps the first report on ASE from double excimer of the conjugated polymer, MEH-PPV in liquid solution

Antigen targeting to dendritic cells elicits long-lived T cell help for antibody responses

Resistance to several prevalent infectious diseases requires both cellular and humoral immune responses. T cell immunity is initiated by mature dendritic cells (DCs) in lymphoid organs, whereas humoral responses to most antigens require further collaboration between primed, antigen-specific helper T cells and naive or memory B cells. To determine whether antigens delivered to DCs in lymphoid organs induce T cell help for antibody responses, we targeted a carrier protein, ovalbumin (OVA), to DCs in the presence of a maturation stimulus and assayed for antibodies to a hapten, (4-hydroxy-3-nitrophenyl) acetyl (NP), after boosting with OVA-NP. A single DC-targeted immunization elicited long-lived T cell helper responses to the carrier protein, leading to large numbers of antibody-secreting cells and high titers of high-affinity antihapten immunoglobulin Gs. Small doses of DC-targeted OVA induced higher titers and a broader spectrum of anti-NP antibody isotypes than large doses of OVA in alum adjuvant. Similar results were obtained when the circumsporozoite protein of Plasmodium yoelii was delivered to DCs. We conclude that antigen targeting to DCs combined with a maturation stimulus produces broad-based and long-lived T cell help for humoral immune responses

Effect of Humic Acid on Seed Germination of Raphanus sativus L

Abstract: In the present study, we have tested the effect of humic acid on seed germination of Radish (Raphanus sativus). Seeds were soaked in various concentrations (0.1%, 0.25%, 0.5%, 0.75% and 1%) of humic acid at different time periods (10, 60, 120, 180 and 240 minutes). After 7 days, the seeds were analysed for their germination capacity, root and shoot length. The study infers that humic acid with the concentration of 0.25% showed maximum seed germination (100%) and the optimum shoot and root length was recorded as 6.175cm and 11.46cm respectively after 60 minutes soaking

Aldosterone and vasopressin affect α- and γ-ENaC mRNA translation

Vasopressin and aldosterone play key roles in the fine adjustment of sodium and water re-absorption in the nephron. The molecular target of this regulation is the epithelial sodium channel (ENaC) consisting of α-, β- and γ-subunits. We investigated mRNA-specific post-transcriptional mechanisms in hormone-dependent expression of ENaC subunits in mouse kidney cortical collecting duct cells. Transcription experiments and polysome gradient analysis demonstrate that both hormones act on transcription and translation. RNA-binding proteins (RBPs) and mRNA sequence motifs involved in translational control of γ-ENaC synthesis were studied. γ-ENaC–mRNA 3′-UTR contains an AU-rich element (ARE), which was shown by RNA affinity chromatography to interact with AU-rich element binding proteins (ARE-BP) like HuR, AUF1 and TTP. Some RBPs co-localized with γ-ENaC mRNA in polysomes in a hormone-dependent manner. Reporter gene co-expression experiments with luciferase γ-ENaC 3′-UTR constructs and ARE-BP expression plasmids demonstrate the importance of RNA–protein interaction for the up-regulation of γ-ENaC synthesis. We document that aldosterone and the V2 receptor agonist dDAVP act on synthesis of α- and γ-ENaC subunits mediated by RBPs as effectors of translation but not by mRNA stabilization. Immunoprecipitation and UV-crosslinking analysis of γ-ENaC–mRNA/HuR complexes document the significance of γ-ENaC–mRNA–3′-UTR/HuR interaction for hormonal control of ENaC synthesis

Clathrin and LRP-1-Independent Constitutive Endocytosis and Recycling of uPAR

Background: The urokinase receptor (uPAR/CD87) is highly expressed in malignant tumours. uPAR, as a GPI anchored protein, is preferentially located at the cell surface, where it interacts with its ligands urokinase (uPA) and the extracellular matrix protein vitronectin, thus promoting plasmin generation, cell-matrix interactions and intracellular signalling events. Interaction with a complex formed by uPA and its inhibitor PAI-1 induces cell surface down regulation and recycling of the receptor via the clathrin-coated pathway, a process dependent on the association to LRP-1. Methodology/Principal Findings: In this study, we have found that along with the ligand-induced down-regulation, uPAR also internalizes and recycles constitutively through a second pathway that is independent of LRP-1 and clathrin but shares some properties with macropinocytosis. The ligand-independent route is amiloride-sensitive, does not require uPAR partitioning into lipid rafts, is independent of the activity of small GTPases RhoA, Rac1 and Cdc42, and does not require PI3K activity. Constitutively endocytosed uPAR is found in EEA1 positive early/recycling endosomes but does not reach lysosomes in the absence of ligands. Electron microscopy analysis reveals the presence of uPAR in ruffling domains at the cell surface, in macropinosome-like vesicles and in endosomal compartments. Conclusions/Significance: These results indicate that, in addition to the ligand-induced endocytosis of uPAR, efficient surface expression and membrane trafficking might also be driven by an uncommon macropinocytic mechanism couple