Biochemical analysis on crop shoots of Camellia sinensis (L.) O. Kuntze tea from the selected UPASI-16 clone.

Commercial tea comes from plants belonging to a relatively large group of cultivated species of Camellia sinensis (L) O. Kuntze. It is one of the oldest known beverages made from the tender leaves of the plants. The main aim of the present study is to analyze the presence of biochemicals in UPASI -16 clone. In the results, the UPASI -16 clone showed high catechin (22.88%), polyphenol (31.70%), polyphenol oxidase (1178.63 U/mg of protein) and tea enzyme peroxidase (729.72 µM of O2 formed min-1g-1 dry weight), catalase (1.89 µM H2O2 reduced min-1 mg-1 protein) and Super oxide dismutase (58.45 U/mg proteins). UPASI -16 showed high catechin content in segregated tea crop shoots from the first internodes (23.66%) followed by the first leaf (22.46%), second leaf and third leaf.  Tea quality flavonoid gene expression, while ANR (1.66%) and F3H (1.02%) were down regulated, F35H and ANS (2.82%) were up regulated in UPASI -16. Based on the results, it can be concluded that crop shoots of Camellia sinensis (L) O. Kuntze selected clone of UPASI-16 may possess high amount of biochemicals and may further lead to development of commercial tea

Molecular inversion probe-based SPR biosensing for specific, label-free and real-time detection of regional DNA methylation

DNA methylation has the potential to be a clinically important biomarker in cancer. This communication reports a real-time and label-free biosensing strategy for DNA methylation detection in the cancer cell line. This has been achieved by using surface plasmon resonance biosensing combined with the highly specific molecular inversion probe based amplification method, which requires only 50 ng of bisulfite treated genomic DNA

A Patient-centric, Attribute-based, Source-verifiable Framework for Health Record Sharing

The storage of health records in electronic format, and the wide-spread sharing of these records among different health care providers, have enormous potential benefits to the U.S. healthcare system. These benefits include both improving the quality of health care delivered to patients and reducing the costs of delivering that care. However, maintaining the security of electronic health record systems and the privacy of the information they contain is paramount to ensure that patients have confidence in the use of such systems. In this paper, we propose a framework for electronic health record sharing that is patient centric, i.e. it provides patients with substantial control over how their information is shared and with whom; provides for verifiability of original sources of health information and the integrity of the data; and permits fine-grained decisions about when data can be shared based on the use of attribute-based techniques for authorization and access control. We present the architecture of the framework, describe a prototype system we have built based on it, and demonstrate its use within a scenario involving emergency responders' access to health record information

Accurate detection of methylated cytosine in complex methylation landscapes

Monitoring DNA methylation can be a useful biomarker for disease diagnosis and prognosis. However, monitoring the methylation status of a specific cytosine biomarker is often confounded by heterogeneous peripheral DNA methylation. To address this issue, molecular inversion probes were designed with inosine strategically positioned to complement suspected DNA methylation sites. This enabled the methylation status of a specific cytosine to be accurately measured with a high level of specificity, irrespective of adjacent epigenetic modifications

Considerations of solid-phase DNA amplification

Solid-phase (SP) polymerase chain reaction (PCR) is an increasingly popular tool used to produce immobilized DNA for a variety of applications, including high-throughput DNA sequencing and SNP analysis. Despite its usefulness, the mechanism of DNA amplification using immobilized primers has not been thoroughly explored. Herein, we describe a SP-PCR process that was designed to explore and better understand some limitations of SP-DNA amplification. The rate of SP-DNA amplification was measured, and the ability to exponentially amplify DNA on a surface was demonstrated. Approximately 50 amol of DNA was amplified to detectable levels using SP-PCR. The mechanism and some limitations of the reaction were investigated by measuring the density of the primer on the surface prior to amplification and the amount of immobilized amplicon produced after SP-PCR. This enabled some of the practical limitations of the reaction to be addressed within a logical theoretical framework

Molecular Inversion Probe: A New Tool for Highly Specific Detection of Plant Pathogens

<div><p>Highly specific detection methods, capable of reliably identifying plant pathogens are crucial in plant disease management strategies to reduce losses in agriculture by preventing the spread of diseases. We describe a novel molecular inversion probe (MIP) assay that can be potentially developed into a robust multiplex platform to detect and identify plant pathogens. A MIP has been designed for the plant pathogenic fungus <i>Fusarium oxysporum</i> f.sp. <i>conglutinans</i> and the proof of concept for the efficiency of this technology is provided. We demonstrate that this methodology can detect as little as 2.5 ng of pathogen DNA and is highly specific, being able to accurately differentiate <i>Fusarium oxysporum</i> f.sp. <i>conglutinans</i> from other fungal pathogens such as <i>Botrytis cinerea</i> and even pathogens of the same species such as <i>Fusarium oxysporum</i> f.sp. <i>lycopersici</i>. The MIP assay was able to detect the presence of the pathogen in infected <i>Arabidopsis thaliana</i> plants as soon as the tissues contained minimal amounts of pathogen. MIP methods are intrinsically highly multiplexable and future development of specific MIPs could lead to the establishment of a diagnostic method that could potentially screen infected plants for hundreds of pathogens in a single assay.</p></div

Molecular inversion probe-based SPR biosensing for specific, label-free and real-time detection of regional DNA methylation

DNA methylation has the potential to be a clinically important biomarker in cancer. This communication reports a real-time and label-free biosensing strategy for DNA methylation detection in the cancer cell line. This has been achieved by using surface plasmon resonance biosensing combined with the highly specific molecular inversion probe based amplification method, which requires only 50 ng of bisulfite treated genomic DNA.This work was supported by the UQ fellowship (2012001456) awarded to LGC, ARC DECRA (DE120102503) awarded to MJAS, and National Breast Cancer Foundation (NBCF) of Australia (CG-08-07 and CG-12-07) awarded to MT.Peer Reviewe

Methylsorb: A Simple Method for Quantifying DNA Methylation Using DNA-gold Affinity Interactions

The analysis of DNA methylation is becoming increasingly important both in the clinic and also as a research tool to unravel key epigenetic molecular mechanisms in biology. Current methodologies for the quantification of regional DNA methylation are largely affected by comprehensive DNA sequencing methodologies which tend to be expensive, tedious, and time-consuming for many applications. Herein, we report an entirely new DNA methylation detection method referred to as 'Methylsorb', which is based on the inherent affinity of DNA bases to the gold surface (i.e., the trend of the affinity interactions is adenine > cytosine ≥ guanine > thymine) [1]. Since the degree of gold-DNA affinity interaction is highly sequence-dependent, it provides a new capability to detect DNA methylation by simply monitoring the relative adsorption of bisulfite treated DNA sequences onto a gold chip. Because the selective physical adsorption of DNA fragments to gold enable a direct read-out of regional DNA methylation, the current requirement for DNA sequencing is obviated. To demonstrate the utility of this method we present data on the regional methylation status of two CpG clusters located in the EN1 and MIR200B genes in MCF7 and MDA-MB-231 cells. The methylation status of these regions was obtained from the change in relative mass on gold surface with respect to relative adsorption of an unmethylated DNA source and this was detected using surface Plasmon resonance (SPR) in a label-free and real-time manner. We anticipate that the simplicity of this method, combined with the high level of accuracy for identifying the methylation status of cytosines in DNA, could find broad application in biology and diagnostics

Oligonucleotide sequences.

<p>Oligonucleotide sequences.</p