Improved protocol for plasma microRNA extraction and comparison of commercial kits

IntroductionMicroRNAs are small, non-coding RNA molecules that are becoming popular biomarkers in several diseases. However, their low abundance in serum/plasma poses a challenge in exploiting their potential in clinics. Several commercial kits are available for rapid isolation of microRNA from plasma. However, reports guiding the selection of appropriate kits to study downstream assays are scarce. Hence, we compared four commercial kits to evaluate microRNA-extraction from plasma and provided a modified protocol that further improved the superior kit’s performance.Materials and methodsWe compared four kits (miRNeasy Serum/Plasma, miRNeasy Mini Kit from Qiagen; RNA-isolation, and Absolutely-RNA MicroRNA Kit from Agilent technologies) for quality and quantity of microRNA isolated, extraction efficiency, and cost-effectiveness. Bioanalyzer-based Agilent Small RNA kit was used to evaluate quality and quantity of microRNA. Extraction efficiency was evaluated by detection of four endogenous control microRNA using real-time-PCR. Further, we modified the manufacturer’s protocol for miRNeasy Serum/Plasma kit to improve yield.ResultsmiRNeasy Serum/Plasma kit outperformed the other three kits in microRNA-quality (P < 0.005) and yielded maximum microRNA-quantity. Recovery of endogenous control microRNA i.e. hsa-miR-24-3p, hsa-miR-191-5p, hsa-miR-423-5p and hsa-miR-484 was higher as well. Modification with the inclusion of a double elution step enhanced yield of microRNA extracted with miRNeasy Serum/Plasma kit significantly (P < 0.001).ConclusionWe demonstrated that miRNeasy Serum/Plasma kit outperforms other kits and can be reliably used with a limited plasma quantity. We have provided a modified microRNA-extraction protocol with improved microRNA output for downstream analyses.     

Peasant cultures of the twenty‐first century 1

Abstract

In the last quarter of a century there has been a fundamental change in the historical situation of post‐coloniality. The new conditions under which global flows of capital, commodities, information and people are now regulated have created both new opportunities and new obstacles for post‐colonial countries. The old idea of a Third World, sharing a common history of colonial oppression and backwardness, is no longer as persuasive as it was in the 1960s. The phenomenal growth of China and India in recent years has set in motion a process of social change that, in its scale and speed, is unprecedented in human history. I will argue that the forms of capitalist industrial growth in the twenty‐first century may, in large agrarian countries like China, India and the countries of South‐east Asia, make room for the preservation of peasant production and peasant cultures, but under completely altered conditions. The analysis of these emergent forms of postcolonial capitalism requires new conceptual work.     

Theory of Dielectric Optical Waveguides,by Dietrich Marcuse. 275 pages,diagrams, illustrations, 612× 912in. New York,Academic Press Inc., 1974. Price, $21.00.

A thin shell model of the uterus is proposed and a kinematic solution of uterine deformation during labor derived. In vivo multi-point strain measurements performed during the various stages of labor up to delivery form boundary conditions for the numerical solution, which yields curvature and local obliqueness variations of the uterus during a contraction. The results are discussed in the light of their clinical applications.     

Total Serum Immunoglobulin E in Children with Asthma

Immunoglobulin (Ig) E has been shown to be a major contributing factor for the development of bronchial hyperresponsiveness in asthma. An elevation in serum IgE levels contributes to asthma and is considered a potent predictor of the development of asthma. The objectives of the present study were to estimate the levels of total serum IgE in asthmatic and healthy control subjects and to investigate the relationship of various demographic and clinical characteristics with the total serum IgE level in asthmatics. We measured the levels of total serum IgE using the ELISA kits (AccuBind, Monobind Inc., USA). The relevant demographic and clinical data were obtained using the questionnaire. The results showed that asthmatic children had significantly elevated level of total serum IgE compared to that of the healthy controls. The levels of total IgE and IL-4 in sera of 44 asthmatic children showed a significant positive correlation. Total serum IgE >150 IU/mL was found to be significantly associated with the age, exposure to cigarette smoke, and raised eosinophil count in asthmatic children. In conclusion, the elevated level of total serum IgE may demonstrate the allergic etiology of asthma in the subjects studied.     

The Brain on Music

Permeating across societies and cultures, music is a companion to millions across the globe. Despite being an abstract art form, music affects all of us deeply, encouraging us in periods of difficulty, comforting us in moments of sadness and celebrating with us in times of joy. In recent years, neuroscientists have recognized that listening to and producing music involves a tantalizing mix of practically every human cognitive function. Thus, music has emerged as an invaluable tool to study varying aspects of the human brain such as auditory and motor perception and learning, attention, memory, and emotion. In this paper, we will discuss the different attributes of music and the neural structures associated with them. We will also highlight some recent work in our laboratory at the National Brain Research Centre in India on emotions associated with Hindustani rāga music.     

Statistics and Intuition for the Classroom

Examples and case studies are presented of statistical reasoning, thinking and intuition that may arise in perception of randomness and in particular for random walks. The relationship between art and science can be explored through various notions of the statistical concepts of randomness.