Chemical Synthesis of Sensitive DNA

Over the past decades, researchers have tried various chemical methods to synthesize modified oligodeoxynucleotides (ODNs, i.e. short segments of DNAs). Traditional ODN synthesis methods require strong basic, and nucleophilic conditions for the deprotection and cleavage of the ODN from the solid support. However, the sensitive ODNs containing labile functionalities are vulnerable to such harsh conditions. Sensitive ODNs have a wide range of applications in research and pharmaceuticals. To synthesize sensitive ODNs, researchers devised different strategies but no practical methods have been developed. To overcome these challenges, we developed alkyl Dim alkyl Dmoc technology. This innovative technology uses weakly basic and non-nucleophilic conditions to synthesize the sensitive ODNs. To demonstrate the feasibility of alkyl Dim alkyl Dmoc technology different ODNs were synthesized, purified using RP-HPLC, and analyzed using MALDI-TOF MS and capillary electrophoresis. Using the method, we successfully synthesized various sensitive ODNs that cannot be synthesized or are highly challenging to synthesize using any known methods. The longest ODN synthesized using methyl Dmoc technology was 23-mer. To further extend the utility and capability of alkyl Dim alkyl Dmoc technology, we aimed to synthesize longer sensitive ODNs. To achieve this goal, PEGylated Dmoc (pDmoc) technology was developed. The introduction of the PEG Dmoc group increased the ODN solubility which facilitated the synthesis of longer ODNs. The longest ODN synthesized was a 49-mer. Moreover, a sensitive 4acC epigenetically modified ODN was synthesized. The success of synthesizing this naturally occurring sensitive ODN will make many projects in the area of epigenetics feasible

A novel optimized deep learning method for protein-protein prediction in bioinformatics

Proteins have been shown to perform critical activities in cellular processes and are required for the organism's existence and proliferation. On complicated protein-protein interaction (PPI) networks, conventional centrality approaches perform poorly. Machine learning algorithms based on enormous amounts of data do not make use of biological information's temporal and spatial dimensions. As a result, we developed a sequence-dependent PPI prediction model using an Aquila and shark noses-based hybrid prediction technique. This model operates in two stages: feature extraction and prediction. The features are acquired using the semantic similarity technique for good results. The acquired features are utilized to predict the PPI using hybrid deep networks long short-term memory (LSTM) networks and restricted Boltzmann machines (RBMs). The weighting parameters of these neural networks (NNs) were changed using a novel optimization approach hybrid of aquila and shark noses (ASN), and the results revealed that our proposed ASN-based PPI prediction is more accurate and efficient than other existing techniques

Improved Parallel Rabin-Karp Algorithm Using Compute Unified Device Architecture

String matching algorithms are among one of the most widely used algorithms in computer science. Traditional string matching algorithms efficiency of underlaying string matching algorithm will greatly increase the efficiency of any application. In recent years, Graphics processing units are emerged as highly parallel processor. They out perform best of the central processing units in scientific computation power. By combining recent advancement in graphics processing units with string matching algorithms will allows to speed up process of string matching. In this paper we proposed modified parallel version of Rabin-Karp algorithm using graphics processing unit. Based on that, result of CPU as well as parallel GPU implementations are compared for evaluating effect of varying number of threads, cores, file size as well as pattern size.Comment: Information and Communication Technology for Intelligent Systems (ICTIS 2017

Sexual Offender Treatment: A Paradigm Analysis of Academic Journals

Many criminologists and psychologists have theorized the possible causes behind an individual who engages in sex offenses; some of which will be reviewed. Through this paradigm analysis, I hope to identify how each discipline addresses the causes of sex offending and what treatments they offer in response to their theories. Additionally, this thesis will examine the Good Lives, Relapse Prevention, and the Risks, Need, Responsivity (RNR) models of treatment. The main objective of this thesis is to address the different facets of sex offending, so that the importance of finding an effective treatment model can be understood. By raising awareness to the multiple typologies of sex offenders and differentiating contact and non-contact offenses, this thesis aims to allow for a better understanding of the causes of sexual offending so that we may develop effective treatment options that address such issues

Determination of Optical Density (OD) of Oligodeoxynucleotide from HPLC Peak Area

Oligodeoxynucleotides (ODNs) are typically purified and analysed with HPLC equipped with a UV-Vis detector. Quantities of ODNs are usually determined using a UV-Vis spectrometer separately after HPLC, and are reported as optical density at 260 nm (OD260). Here, we describe a method for direct determination of OD260 of ODNs using the area of the peaks in HPLC profiles

Long oligodeoxynucleotides: chemical synthesis, isolation via catching-by-polymerization, verification via sequencing, and gene expression demonstration

Long oligodeoxynucleotides (ODNs) are segments of DNAs having over one hundred nucleotides (nt). They are typically assembled using enzymatic methods such as PCR and ligation from shorter 20 to 60 nt ODNs produced by automated de novo chemical synthesis. While these methods have made many projects in areas such as synthetic biology and protein engineering possible, they have various drawbacks. For example, they cannot produce genes and genomes with long repeats and have difficulty to produce sequences containing stable secondary structures. Here, we report a direct de novo chemical synthesis of 400 nt ODNs, and their isolation from the complex reaction mixture using the catching-by-polymerization (CBP) method. To determine the authenticity of the ODNs, 399 and 401 nt ODNs were synthesized and purified with CBP. The two were joined together using Gibson assembly to give the 800 nt green fluorescent protein (GFP) gene construct. The sequence of the construct was verified via Sanger sequencing. To demonstrate the potential use of the long ODN synthesis method, the GFP gene was expressed in E. coli. The long ODN synthesis and isolation method presented here provides a pathway to the production of genes and genomes containing long repeats or stable secondary structures that cannot be produced or are highly challenging to produce using existing technologies

Measuring Procedural Justice Policy Adherence During Use of Force Events: The Body-Worn Camera as a Performance Monitoring Tool

This study capitalizes on a successful researcher–practitioner partnership to conduct a systematic social observation (SSO) of police body-worn camera (BWC) footage in Newark, NJ. To demonstrate the utility of BWCs as performance monitoring tools, we measure officer adherence to procedural justice standards throughout use of force events as mandated in the Newark Police Division’s updated policies pursuant to an ongoing federal consent decree. Overall, a slim majority of use of force events are procedurally just. However, results indicate several instances of policy noncompliance. Results are discussed, and policy recommendations related to procedural justice policy violations and BWCs for performance monitoring are provided

An agonist sensitive, quick and simple cell-based signaling assay for determination of ligands mimicking prostaglandin E2 or E1 activity through subtype EP1 receptor: Suitable for high throughput screening

<p>Abstract</p> <p>Background</p> <p>Conventionally the active ingredients in herbal extracts are separated into individual components, by fractionation, desalting, and followed by high-performance liquid chromatography (HPLC). In this study we have tried to directly screen water-soluble fractions of herbs with potential active ingredients before purification or extraction. We propose that the herbal extracts mimicking prostaglandin E₁(PGE₁) and E₂(PGE₂) can be identified in the water-soluble non-purified fraction. PGE₁is a potent anti-inflammatory molecule used for treating peripheral vascular diseases while PGE₂is an inflammatory molecule.</p> <p>Methods</p> <p>We used cell-based assays (CytoFluor multi-well plate reader and fluorescence microscopy) in which a calcium signal was generated by the recombinant EP₁receptor stably expressed in HEK293 cells (human embryonic kidney). PGE₁and PGE₂were tested for their ability to generate a calcium signal. Ninety-six water soluble fractions of Treasures of the east (single Chinese herb dietary supplements) were screened.</p> <p>Results</p> <p>After screening, the top ten stimulators were identified. The identified herbs were then desalted and the calcium fluorescent signal reconfirmed using fluorescence microscopy. Among these top ten agonists identified, seven stimulated the calcium signaling (1-40 μM concentration) using fluorescence microscopy.</p> <p>Conclusions</p> <p>Fluorescence microscopy and multi-well plate readers can be used as a target specific method for screening water soluble fractions with active ingredients at a very early stage, before purification. Our future work consists of purifying and separating the active ingredients and repeating fluorescence microscopy. Under ordinary circumstances we would have to purify the compounds first and then test all the extracts from 96 herbs. Conventionally, for screening natural product libraries, the procedure followed is the automated separation of all constituents into individual components using fractionation and high performance liquid chromatography. We, however, demonstrated that the active ingredients of the herbal extracts can be tested before purification using an agonist sensitive, quick and simple cell-based signaling assay for ligands mimicking the agonists, PGE₁and PGE₂.</p

Addressing climate change with behavioral science:A global intervention tournament in 63 countries

Effectively reducing climate change requires marked, global behavior change. However, it is unclear which strategies are most likely to motivate people to change their climate beliefs and behaviors. Here, we tested 11 expert-crowdsourced interventions on four climate mitigation outcomes: beliefs, policy support, information sharing intention, and an effortful tree-planting behavioral task. Across 59,440 participants from 63 countries, the interventions' effectiveness was small, largely limited to nonclimate skeptics, and differed across outcomes: Beliefs were strengthened mostly by decreasing psychological distance (by 2.3%), policy support by writing a letter to a future-generation member (2.6%), information sharing by negative emotion induction (12.1%), and no intervention increased the more effortful behavior-several interventions even reduced tree planting. Last, the effects of each intervention differed depending on people's initial climate beliefs. These findings suggest that the impact of behavioral climate interventions varies across audiences and target behaviors.</p