Novel method for screening pigeonpea varieties for globulin content by radioimmunoassay

Summary A rapid and sensitive radioimmunoassay (RIA) is described for screening a large number of pigeonpea varieties for quantitation of globulin content. This method employs precipitation of iodine-labelled pigeonpea globulin by polyethylene glycol and measuring the labelled antigen-antibody complex. Among the 56 varieties screened, Gwalior-3 was found to contain the highest globulin content. RIA when used to quantitate the pigeonpea globulin levels during post anthesis revealed that maximum globulin is present between 24 to 28 days after flowering.NCL Communication No. 3819.     

Opioid Peptides: An Overview of Functional Significance

International Journal of Peptide Research and Therapeutics - Bioactive peptides have been reported to exhibit opioid-like activity and are termed as opioid peptides. Either these are produced...     

Exploration of highly selective fluorogenic ‘on–off' chemosensor for H2PO4− ions: ICT‐based sensing and ATPase activity profiling

Abstract In this study, the recognition contour of Chemosensor 1 was investigated using semiaqueous methanol (X_H, mole fraction = 0.31) for a range of anions and bioactive species. Host–receptor signalling based on the internal charge transfer mechanism for Chemosensor 1 was explored and reported. Structure of Chemosensor 1 and its plausible anion coordination based on hydrogen bonding is complemented with density functional theory. Consequently, we investigated the applicability of the synthesized probe in blood plasma, urine, tap water samples, and for monitoring of ATP in lysosomes by apyrase enzyme.     

Low oxygen tension potentiates proliferation and stemness but not multilineage differentiation of caprine male germline stem cells

The milieu of male germline stem cells (mGSCs) is characterized as a low-oxygen (O₂) environment, whereas, their in-vitro expansion is typically performed under normoxia (20–21% O₂). The comparative information about the effects of low and normal O₂ levels on the growth and differentiation of caprine mGSCs (cmGSCs) is lacking. Thus, we aimed to investigate the functional and multilineage differentiation characteristics of enriched cmGSCs, when grown under hypoxia and normoxia. After enrichment of cmGSCs through multiple methods (differential platting and Percoll-density gradient centrifugation), the growth characteristics of cells [population-doubling time (PDT), viability, proliferation, and senescence], and expression of key-markers of adhesion (β-integrin and E-Cadherin) and stemness (OCT-4, THY-1 and UCHL-1) were evaluated under hypoxia (5% O₂) and normoxia (21% O₂). Furthermore, the extent of multilineage differentiation (neurogenic, adipogenic, and chondrogenic differentiation) under different culture conditions was assessed. The survival, viability, and proliferation were significantly (p?<?0.05) improved, thus, yielding a significantly (p?<?0.05) higher number of viable cells with larger colonies under hypoxia. Furthermore, the expression of stemness and adhesion markers were distinctly upregulated under lowered O₂ conditions. Conversely, the differentiated regions and expression of differentiation-specific genes [C/EBPα (adipogenic), nestin and β-tubulin (neurogenic), and COL2A1 (chondrogenic)] were significantly (p?<?0.05) reduced under hypoxia. Overall, the results demonstrate that culturing cmGSCs under hypoxia augments the growth characteristics and stemness but not the multilineage differentiation of cmGSCs, as compared with normoxia. These data are important to develop robust methodologies for ex-vivo expansion and lineage-committed differentiation of cmGSCs for clinical applications.

     

Meta-analysis of Genetic Structure and Association of Prolactin Gene with Performance Traits in Dairy Cattle in India

The present meta-analysis was carried to provide the more reliable estimates of gene frequency and association of Rsa 1 generated candidate genotype of prolactin gene within exon-3 with performance traits in 1198 Indian dairy cows using data from 15 published studies. Six genetic models viz., codominant (AA vs. AB, AA vs. BB and AB vs. BB), dominant (AA+AB vs. BB), completely over dominant (AA+BB vs. AB) and recessive (AA vs. AB+BB) were used to obtain standardized mean difference (SMD) between genotypes. Meta-analysis showed that the gene frequency of A allele (156 bp) was 0.60 (95% confidence interval (CI) 0.54, 0.65). In association analysis, cows with AB genotype [SMD?=?0.65, 95% CI 0.00, 1.30] had significantly (P?<?0.05) higher lactation milk yield (LMY) as compared to BB genotype, whereas AA and AB genotypes had similar trend. Likewise, AA?+?AB also had larger effect [SMD?=?2.31, 95% CI 0.21, 4.10] on LMY as compared to BB. Cows with AB genotype had significantly lower age at first calving (AFC) with small effect [SMD (AA vs. AB)?=?1.38, 95% CI 0.06, 2.70] and medium effect [SMD (AB vs. BB)?=????3.83, 95% CI???6.41,???1.24] as compared to cows with AA and BB genotypes, respectively. This finding was confirmed under dominant and completely over dominant models. In case of fat%, AA genotype showed negative effect (SMD?=????0.51, 95% CI???0.84,???0.17) under recessive model. It was concluded that the propagation of allele A is promising to help dairy farmers to improve the genetic quality of their dairy cows.

     

The Synthesis of the Sixteen Possible 2′-O-Methyl MMI Dimer Phosphoramidites: Building Blocks for the Synthesis of Novel Antisense Oligonucleotides

Abstract

The synthesis of Methylene(methylimino) or MMI linked nucleoside dimers in all sixteen possible configurations has been accomplished via a reductive coupling of a nucleosidic aldehyde with an hydroxylamine. This has allowed us to prepare all of the necessary 2′-O-methyl MMI dimer building blocks necessary for use in an antisense motif.     

Density Gradient Multilayered Polymerization (DGMP): A Novel Technique for Creating Multi-compartment,Customizable Scaffolds for Tissue Engineering

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix structure (e.g. composition, concentration, or stiffness of the matrix) vary over space, would enable a wide range of investigations concerning how these variables affect cell differentiation, migration, and other phenomena. The major challenge in creating layered matrices is maintaining the structural integrity of layer interfaces without diffusion of individual components from each layer¹. Current methodologies to achieve this include photopatterning^2-3, lithography⁴, sequential functionalization5, freeze drying⁶, microfluidics⁷, or centrifugation⁸, many of which require sophisticated instrumentation and technical skills. Others rely on sequential attachment of individual layers, which may lead to delamination of layers⁹. DGMP overcomes these issues by using an inert density modifier such as iodixanol to create layers of varying densities¹⁰. Since the density modifier can be mixed with any prepolymer or bioactive molecule, DGMP allows each scaffold layer to be customized. Simply varying the concentration of the density modifier prevents mixing of adjacent layers while they remain aqueous. Subsequent single step polymerization gives rise to a structurally continuous multilayered scaffold, in which each layer has distinct chemical and mechanical properties. The density modifier can be easily removed with sufficient rinsing without perturbation of the individual layers or their components. This technique is therefore well suited for creating hydrogels of various sizes, shapes, and materials.A protocol for fabricating a 2D-polyethylene glycol (PEG) gel, in which alternating layers incorporate RGDS-350, is outlined below. We use PEG because it is biocompatible and inert. RGDS, a cell adhesion peptide¹¹, is used to demonstrate spatial restriction of a biological cue, and the conjugation of a fluorophore (Alexa Fluor 350) enables us to visually distinguish various layers. This procedure can be adapted for other materials (e.g. collagen, hyaluronan, etc.) and can be extended to fabricate 3D gels with some modifications¹⁰.