Functional and Structural Analyses of CYP1B1 Variants Linked to Congenital and Adult-Onset Glaucoma to Investigate the Molecular Basis of These Diseases

Glaucoma, the leading cause of irreversible blindness, appears in various forms. Mutations in CYP1B1 result in primary congenital glaucoma (PCG) by an autosomal recessive mode of inheritance while it acts as a modifier locus for primary open angle glaucoma (POAG). We investigated the molecular basis of the variable phenotypes resulting from the defects in CYP1B1 by using subclones of 23 CYP1B1 mutants reported in glaucoma patients, in a cell based system by measuring the dual activity of the enzyme to metabolize both retinol and 17β-estradiol. Most variants linked to POAG showed low steroid metabolism while null or very high retinol metabolism was observed in variants identified in PCG. We examined the translational turnover rates of mutant proteins after the addition of cycloheximide and observed that the levels of enzyme activity mostly corroborated the translational turnover rate. We performed extensive normal mode analysis and molecular-dynamics-simulationsbased structural analyses and observed significant variation of fluctuation in certain segmental parts of the mutant proteins, especially at the B-C and F-G loops, which were previously shown to affect the dynamic behavior and ligand entry/exit properties of the cytochrome P450 family of proteins. Our molecular study corroborates the structural analysis,and suggests that the pathologic state of the carrier of CYP1B1 mutations is determined by the allelic state of the gene. To our knowledge, this is the first attempt to dissect biological activities of CYP1B1 for correlation with congenital and adult onset glaucomas

Microbe-Based Strategy for Plant Nutrient Management

The rapid industrialization and urbanization of developing countries such as India have encroached on cultivable lands to meet the demands of an ever-increasing population. The altered land use patterns with increased fertilizer use has increased crop yields with leaching of major portion of the applied nutrients from the soil. Nitrates and phosphates are the agricultural pollutants that are discharged into aquifers due to anthropogenic reasons causing severe environmental and health problems. Production of these nutrients requires energy and finite resources (rock phosphate, which has gradually depleting reserves). An alternative management strategy would be to sequester excess nutrients within a biomass that is reused for agriculture. Two discrete enriched microbial consortia with the potential of simultaneous nitrate and phosphate sequestration upon application as biofertilizer restricted them within the plant root zone, ensuring prevention of eutrophication through leaching while making it available for uptake by plants. The nutrient accumulated biomass enhanced the crop yield by 21.88% during mung bean cultivation with maintained elemental content and other nutritional qualities. The major drawback of conventional biofertilizer application (slow release and action) could be overcome using this formulation leading to environmental protection, crop yield enhancement and soil fertility maintenance post-cultivation

A Brief Review on the Regulatory Roles of MicroRNAs in Cystic Diseases and Their Use as Potential Biomarkers

miRNAs are small endogenous conserved non-coding RNA molecules that regulate post-transcriptional gene expression through mRNA degradation or translational inhibition, modulating nearly 60% of human genes. Cystic diseases are characterized by the presence of abnormal fluid-filled sacs in the body, and though most cysts are benign, they can grow inside tumors and turn malignant. Recent evidence has revealed that the aberrant expression of a number of miRNAs present in extracellular fluids, including plasma or serum, urine, saliva, follicular fluid, and semen, contribute to different cystic pathologies. This review aims to describe the role of different miRNAs in three worldwide relevant cystic diseases: polycystic ovarian syndrome (PCOS), polycystic kidney disease (PKD), and pancreatic cyst tumors (PCTs), as well as their potential use as novel biomarkers.publishedVersio

Measurement of tacit knowledge and its conversion to explicit knowledge: a case study of Bolpur college library

113-119A questionnaire survey was conducted to collect user feedback on various elements of library administration as well as users’ expectations to improve library services. A construct model has been deduced using exploratory factor analysis (EFA) by identifying optimal number of significant factors. The proposed construct has been validated by using structural equation modeling (SEM) in terms of convergent and discriminant validities. The proposed framework has been implemented on a survey of the users of Bolpur college library. The results indicate effectiveness in analyzing the TK and its conversion to explicit knowledge and has been proven to be a positive step towards improvement of library services

Measurement of tacit knowledge and its conversion to explicit knowledge: a case study of Bolpur college library

A questionnaire survey was conducted to collect user feedback on various elements of library administration as well as users’ expectations to improve library services. A construct model has been deduced using exploratory factor analysis (EFA) by identifying optimal number of significant factors. The proposed construct has been validated by using structural equation modeling (SEM) in terms of convergent and discriminant validities. The proposed framework has been implemented on a survey of the users of Bolpur college library. The results indicate effectiveness in analyzing the TK and its conversion to explicit knowledge and has been proven to be a positive step towards improvement of library services

MD simulation analysis of the R117P and wild type CYP1B1 structures.

<p><b><i>Panel A</i></b> shows a multiple sequence alignment of CYP1B1 homologs. The red color indicates the conserved Arginine 117 position in other homologs. Thelower panel shows the structural importance of Arginine 117in their interaction with the O1A/O2A atom of the heme ligand. <b><i>Panel B</i></b> shows a similar distribution of first three major principal components of R117P and wild type (WT) CYP1B1 structures. <i>Panel B</i> shows that the R117P mutant possesses a significantly altered flexibility pattern within the B-C, G-H, and J-K block regions. <b><i>Panel C</i></b> shows the altered flexibility pattern in the R117P mutant as compared to WT. The log₂ ratio is calculated as . Therefore, a positive value indicates an increase in flexibility in the R117P mutant and a negative value indicates a decrease in flexibility as compared to WT CYP1B1. The R117P mutant has a significantly altered flexibility pattern within the B-C, G-H and J-K block regions, shown separately in the right hand side of the panel. <b><i>Panels D</i> and <i>E</i></b> illustrate RMSD deviation and average bond angle deviation of the heme ligand in ΔRMSD and Δdegrees matrices, respectively. The difference matrices were calculated by subtracting RMSD and average bond angle values of mutant CYP1B1 from that of WT CYP1B1. The fluctuations and bond angle deviations in the initial stages of the MD simulation indicate a potential instability in the heme ligand binding affinity within the mutant protein.</p

Summary of possible effects of mutations on CYP1B1 structure observed through MD simulation.

<p>Summary of possible effects of mutations on CYP1B1 structure observed through MD simulation.</p

Genotype to phenotype correlation for the role of <i>CYP1B1</i> in glaucoma pathogenesis (PCG vs POAG).

<p>The flowchart shows the potential activity of CYP1B1 variants for two different substrates (estradiol and retinol), as estimated by an in vitro cell based assay in HEK293T cells and attempted correlation of the biochemical activities (based on genotype) with potential glaucoma pathogenesis. Ref.⁽¹⁾[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref018" target="_blank">18</a>]; Ref.^(2,3)[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref092" target="_blank">92</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref093" target="_blank">93</a>]; Ref.⁽⁴⁾[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref017" target="_blank">17</a>]; Ref.^(5,6)[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref017" target="_blank">17</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156252#pone.0156252.ref083" target="_blank">83</a>].</p