Activity of glucose oxidase functionalized onto magnetic nanoparticles

BACKGROUND: Magnetic nanoparticles have been significantly used for coupling with biomolecules, due to their unique properties. METHODS: Magnetic nanoparticles were synthesized by thermal co-precipitation of ferric and ferrous chloride using two different base solutions. Glucose oxidase was bound to the particles by direct attachment via carbodiimide activation or by thiophene acetylation of magnetic nanoparticles. Transmission electron microscopy was used to characterize the size and structure of the particles while the binding of glucose oxidase to the particles was confirmed using Fourier transform infrared spectroscopy. RESULTS: The direct binding of glucose oxidase via carbodiimide activity was found to be more effective, resulting in bound enzyme efficiencies between 94–100% while thiophene acetylation was 66–72% efficient. Kinetic and stability studies showed that the enzyme activity was more preserved upon binding onto the nanoparticles when subjected to thermal and various pH conditions. The overall activity of glucose oxidase was improved when bound to magnetic nanoparticles CONCLUSION: Binding of enzyme onto magnetic nanoparticles via carbodiimide activation is a very efficient method for developing bioconjugates for biological application

Isozyme Analysis on Different Varieties of Sugarcane

Isozymic and protein diversity among five sugarcane varieties viz., Co 6304, Co 85019, Co 8371, Co 89003 and Co 91010 were studied to understand the varietal interrelationship and to identify the biochemical marker for the disease resistance and stress tolerance. The standard technique of vertical gel electrophoresis PAGE was employed for size separation of isozymes. The gel was stained with different staining solutions for different isozyme systems viz. peroxidase, esterase, acid phosphatase, alkaline phosphatase and proteins. Rf values of the banding profiles, similarity index and variation between the varieties were analysed. Among the four enzyme systems, peroxidase profile reveals the difference between the disease resistant / susceptible and abiotic stress tolerant / non tolerant varieties. The two isoperoxidase bands with Rf values 0.62 and 0.66 showed their presence in disease resistant and abiotic tolerant varieties. The presence of two marker bands (0.62, 0.66) of resistant and stress tolerant varieties suggest that the variety Co 6304 may also be resistant to smut, wilt and moderately resistant to red rot and tolerant to drought

How to Identify Rare and Endangered Ferns and Fern Allies

Identification of rare and endangered plant species is the first requirement for any conservation programme. The IUCN guideline is the only available method to identify the rare and endangered species and it requires vast data on the wild population of the target species. None of the biological characters, which are playing main role in the survival and distribution of several species, is used in IUCN guideline. In the meantime there are several difficulties in following IUCN guideline, particularly the non availability of complete field data. Moreover, the same guideline can not be used for all the groups of species in equal importance. The vascular cryptogams, pteridophytes, are also an important component of any mountainous flora and they have also to be conserved in nature. As they are the primitive vascular plants on the earth, they are getting depleted in the flora due to various reasons and it is the right time to identify the rare and endangered pteridophytes to conserve them. By considering various difficulties of IUCN method for the identification rare and endangered pteridophytes, a very simple method has been adopted by using just four criteria and this method can be applied to Pteridophytes from any region of the world

Quantification of 5-methylcytosine, 5-hydroxymethylcytosine and 5-carboxylcytosine from the blood of cancer patients by an Enzyme-based Immunoassay

BACKGROUND: Genome-wide aberrations of the classic epigenetic modification 5-methylcytosine (5mC), considered the hallmark of gene silencing, has been implicated to play a pivotal role in mediating carcinogenic transformation of healthy cells. Recently, three epigenetic marks derived from enzymatic oxidization of 5mC namely 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), have been discovered in the mammalian genome. Growing evidence suggests that these novel bases possess unique regulatory functions and may play critical roles in carcinogenesis. METHODS: To provide a quantitative basis for these rare epigenetic marks, we have designed a biotin-avidin mediated enzyme-based immunoassay (EIA) and evaluated its performance in genomic DNA isolated from blood of patients diagnosed with metastatic forms of lung, pancreatic and bladder cancer, as well as healthy controls. The proposed EIA incorporates spatially optimized biotinylated antibody and a high degree of horseradish-peroxidase (HRP) labeled streptavidin, facilitating signal amplification and sensitive detection. RESULTS: We report that the percentages of 5mC, 5hmC and 5caC present in the genomic DNA of blood in healthy controls as 1.025±0.081, 0.023±0.006 and 0.001±0.0002, respectively. We observed a significant (p<0.05) decrease in the mean global percentage of 5hmC in blood of patients with malignant lung cancer (0.013±0.003%) in comparison to healthy controls. CONCLUSION: The precise biological roles of these epigenetic modifications in cancers are still unknown but in the past two years it has become evident that the global 5hmC content is drastically reduced in a variety of cancers. To the best of our knowledge, this is the first report of decreased 5hmC content in the blood of metastatic lung cancer patients and the clinical utility of this observation needs to be further validated in larger sample datasets

Epigenetic Editing of Ascl1 Gene in Neural Stem Cells by Optogenetics

Enzymes involved in epigenetic processes such as methyltransferases or demethylases are becoming highly utilized for their persistent DNA or histone modifying efficacy. Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1), a candidate proneuron gene. Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing. A differentially methylated region at the Ascl1 promoter, isolated from murine dorsal root ganglion (hypermethylated) and striated cells (hypomethylated), was targeted with these optogenetic-epigenetic constructs. Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter. We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity. This proof of concept developed herein holds immense promise for the ability to regulate gene activity via epigenetic modulation with spatiotemporal precision

Effect of Heavy metal stress on spore germination of Pteris confusa T. G. Walker and Pteris argyraea T. Moore

Background: Plants have both constitutive and adaptive mechanisms for coping with the elevated metal concentrations and they are utilized to clean the polluted soil and water. Unlike angiosperms hyperaccumulators, fern hyperaccumulators are equipped with inherent biological characteristics that could be exploited in the phytoremediation strategies aimed at decontaminating polluted sites. Fern spores can be successfully used to screen the hyperaccumualting ferns and also to test the toxicity of the metal contaminated samples. Purpose of the Study: In the present study, a preliminary attempt was made to compare the tolerance capacity of the spores of two ferns; Pteris confusa T. G. Walker and Pteris argyraea T. Moore against the heavy metal zinc (Zinc sulphate). Spores of the two ferns were cultured in Knop’s liquid medium with various concentrations of zinc sulphate (0-200ppm). Results: In the case of P. confusa normal germination was observed in control, 120 ppm and 140 ppm and the germination of spores were failed in 160, 180 and 200 ppm of zinc supplemented cultures. In contrary, P. argyraea showed maximum percentage of spore germination in 140 ppm zinc supplemented cultures and the control and 120 ppm zinc sulphate supplemented cultures were failed to show the germination. The germination percentage and growth rate was decreased in high concentration of zinc sulphate. Rhizoids are showed more tolerance to heavy metal than protonema of P. argyraea. Conclusion: Difference in response of spores to the heavy metal zinc may be due the difference in the hyper-accumulating capacity of the ferns

PVT Property Measurements for Ethyl Propionate, Ethyl Butyrate, and Ethyl Pentanoate Esters from (298 to 393) K and up to 35 MPa

The results of PVT measurements of the liquid phase within the temperature range (298 to 393) K and up to 35 MPa are presented for some aliphatic esters. Measurements were made by means of a vibrating tube densimeter, model DMA 512P from Anton Parr. The calibration of the densimeter was performed with water and n-heptane as reference fluids. The experimental PVT data have been correlated by a Tait equation. This equation gives good results when used to predict the density of the esters using the method proposed by Thomson et al. Isothermal compressibilities, isobaric expansivities, thermal pressure coefficients, and changes in the isobaric heat capacity have been calculated from the Tait equation

Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy

Fluorescence-based single molecule techniques to interrogate gene expression in tissues present a very low signal-to-noise ratio due to the strong autofluorescence and other background signals from tissue sections. This report presents a background-free method using second-harmonic generation (SHG) nanocrystals as probes to quantify the messenger RNA (mRNA) of human epidermal growth receptor 2 (Her2) at single molecule resolution in specific phenotypes at single-cell resolution directly in tissues. Coherent SHG emission from individual barium titanium oxide (BTO) nanoprobes was demonstrated, allowing for a stable signal beyond the autofluorescence window. Her2 surface marker and Her2 mRNA were specifically labeled with BTO probes, and Her2 mRNA was quantified at single copy sensitivity in Her2 expressing phenotypes directly in cancer tissues. Our approach provides the first proof of concept of a cross-platform strategy to probe tissues at single-cell resolution in situ

Reading The Bible, Ecologically: Re-Imagining Our Theological Hermeneutics

Commencing with a personal prelude to ecological biblical hermeneutics and inspired by a prophetic call of Pope Francis, the present paper outlines how in times of profound crisis, the need to choose what counts is urgent. With recourse to the same model of crisis and choice, a brief overview of some of the salient eco-biblical proposals is presented which then sets the stage for the author’s eco-hermeneutical reading of Isaiah 34-35, which brings to the fore the prophetic portrayal wherein otherwise marginal/ized earth-companions meaningfully show similarity with the cherished Israelite traditions and theological visions

Daughter Zions Contemporary Sibling Dalit Assertions vis--vis a Postcolonial Reading of the Book of Lamentations

Taking a cue from a perceptive and a widely-received work of Carleen Mandalfo (Daughter Zion Talks Back to the Prophets: A Dialogic Theology of the Book of Lamentations), the present paper aims to glean some of the salient multi-disciplinary insights of the author. The same insights are grouped around some of the salient postcolonial features that R. S. Sugirtharajah outlines, which together constitute postcolonial construal with attendant illustrations. Then, the paper shifts the spotlight on commensurate features in contemporary Dalit assertions from the Indian subcontinent in order to propose that the latter can be viewed as a meaningful, contemporary sibling of Daughter Zion. Despite the chronological and cultural distances between the Dalits and Daughter Zion of Lamentations, it is proposed here that the assertions of these two likewise siblings may profitably be appropriated by todays readers of courage and commitment in order to come to terms with the angsts and outbursts of people, particularly of those who reel under immense pain and fissure