Elucidating The Diagnostic And Therapeutic Potential Of An RNA Aptamer Against Human Pituitary Tumour Transforming Gene 1

Nucleic acid aptamers are a class of molecular recognition elements capable of target interaction at high specificity and affinity. This study was aimed to generate an RNA aptamer against the Human Pituitary Tumour Transforming Gene 1 (PTTG1) oncoprotein which can serve as a potential diagnostic and therapeutic tool. PTTG1 gene was first cloned into the pET14b vector to incorporate the small 6X Histidine tag. Protein expression and purification ensued with the E. coli Rosetta 2(DE3)pLysS strain under the urea denaturing method. The protein identity was confirmed by western blot using monoclonal anti-PTTG1 antibody. Intrinsic florescence spectra analysis and GST-pulldown interaction analysis with GST-p53 conferred signs of successful protein refolding. 11 SELEX cycles with dual partitioning strategy were performed with positive indication of enrichment. Sequencing of the cycle 8 pool revealed a putative aptamer (SECURA-3) which depicted binding to PTTG1. SECURA-3 had a dissociation constant estimated at 16.41 ± 6.4 nM. SECURA-3 was employed for development of aptamer-based diagnostic assays namely ALISA, aptamer-based dot blot, aptamer based western blot and aptahistostaining. SECURA-3 was extended with a poly A-tail at the 3’-end to functionally labelled with the biotin by hybridization with the biotin labelled oligo dT sequence. SECURA-3 aptamer demonstrated detection of up to 0.0256 nM PTTG1 in the direct ALISA assay and with the sandwich configuration, SECURA-3 demonstrated detection of PTTG1 up to 3.2 nM with the aptamer as the capturing agent and 16 nM with the aptamer as the detection agent

Reconstructing the germination pathway from the Rafflesia seed transcriptome

Rafflesia (Rafflesiaceae) produces the largest flowers in the world but has no stems, roots, or leaves. It is a holoparasitic angiosperm that derives all its nutrients from its host vine, Tetrastigma. All species are also threatened with extinction, but propagating it has been incredibly challenging. Its germination from seed has never been observed. The thesis aims to reconstruct the germination pathway of Rafflesia from its published seed transcriptome to gain insight into its molecular genetics and understand what germination genes can be stimulated to facilitate infection of its host for ex-situ propagation. The published seed transcriptome of Rafflesia speciosa was annotated. These were then bioinformatically compared to the seed germination pathway genes in the model photosynthetic plant Arabidopsis (Brassicaceae) and in another parasitic plant, Striga (Orobanchaceae), to determine if there are similar genes conserved, especially the Strigolactones-responsive gene which promotes Striga germination in response to hormones called strigolactones. The assembled Rafflesia speciosa seed transcriptome contained more than 123,000 transcripts. Out of more than 123,000 de novo assembled transcripts, 7025 with blast hits were mapped and annotated. Among these, genes involved in abscisic acid, auxin, brassinosteroids, cytokinin, and ethylene signalling were detected in Rafflesia, in common with those in Arabidopsis. Still, no Strigolactones-responsive genes were detected in Rafflesia. This study showed that some germination signalling genes are conserved between Rafflesia and Arabidopsis but not with another parasitic plant. This suggests that strigolactones will not be useful in stimulating Rafflesia germination for propagation attempts

Predicting Soaked CBR Value of Fine Grained Soils Using Index and Compaction Characteristics

California Bearing Ratio (CBR) value is an indicator of subgrade soil strength and is used often for design of flexible pavements. The conventional soaked CBR testing method is expensive and time consuming. To overcome this situation, it is appreciable to predict CBR value of subgrade soil with simple properties of soils such as index properties which include grain size analysis (% Gravel, % Sand, % Fines), Liquid Limit (LL), Plastic Limit (PL), and compaction characteristics; namely Maximum Dry Density (MDD) and Optimum Moisture Content (OMC). The present study aims at developing regression-based models for predicting soaked CBR value for fine-grained subgrade soils in terms of gran size analysis, LL, PL, MDD and OMC

A Suit of Record Normalization Methods, From Naive Ones, Globally Mine a Group of Duplicate Records

The promise of Big Data pivots after tending to a few big data integration challenges, for example, record linkage at scale, continuous data combination, and incorporating Deep Web. Although much work has been directed on these issues, there is restricted work on making a uniform, standard record from a gathering of records comparing to a similar genuine element. We allude to this errand as record normalization. Such a record portrayal, instituted normalized record, is significant for both front-end and back-end applications. In this paper, we formalize the record normalization issue, present top to bottom examination of normalization granularity levels (e.g., record, field, and worth segment) and of normalization structures (e.g., common versus complete). We propose an exhaustive structure for registering the normalized record. The proposed system incorporates a suit of record normalization techniques, from guileless ones, which utilize just the data accumulated from records themselves, to complex methodologies, which all around mine a gathering of copy records before choosing an incentive for a quality of a normalized record

An invariant property of Mahler measure

We exhibit a change of variables that maintains the Mahler measure of a given polynomial. This method leads to the construction of highly non-trivial polynomials with given Mahler measure and settles some conjectural numerical formulas due to Boyd and Brunault.Comment: 12 page

Gasification of Low Ash Partially Composted Dairy Biomass with Enriched Air Mixture

Biomass is one of the renewable and non-conventional energy sources and it includes municipal solid wastes and animal wastes in addition to agricultural residue. Concentrated animal feeding operations produce large quantities of cattle biomass which might result in land and water pollution if left untreated. Different methods are employed to extract the available energy from the cattle biomass (CB) which includes co-firing and gasification. There are two types of CB: Feedlot biomass (FB), animal waste from feedlots and dairy biomass (DB), animal waste from dairy farms. Experiments were performed in the part on gasification of both FB and DB. Earlier studies on gasification of DB with different steam-fuel ratios resulted in increased production of hydrogen. In the present study, dairy biomass was gasified in a medium with enriched oxygen percentage varying from 24% to 28%. The effect of enriched air mixture, equivalence ratio and steam-fuel ratio on the performance of gasifier was studied. Limited studies were done using a mixture of carbon dioxide and oxygen as the gasification medium and also a methodology was developed to determine the gasification efficiency based on mass and heat contents of gas. The results show that the peak temperature within the bed increases with increase in oxygen concentration in the gasification medium. Also carbon dioxide concentration in the mixture increases with corresponding decrease in carbon monoxide with increase in oxygen concentration of the incoming gasification medium. The peak temperature increased from 988°C to 1192°C as the oxygen concentration increased from 21% to 28% at ER=2.1. The upper limit on oxygen concentration is limited to 28% due to high peak temperature and resulting ash agglomeration. Higher heating value (HHV) of the gases decreases with increase in equivalence ratio. The gases produced using carbon dioxide and oxygen mixture had a higher HHV when compared to that of air and enriched air gasification. Typically the HHV of the gases increased from 2219 kJ/m³ to 3479 kJ/m³ when carbon dioxide and oxygen mixture is used for gasification instead of air at ER=4.2 in the absence of steam

Synthesis and Bioassay of Azolyl Benzothiazine Carboxamides

Cyclic sulfonamides (sultams) gained importance due to their potential pharmaceutical activity. Amongst these, 1,2-benzothiazine-1,1-dioxide and their derivatives represent the most familiar class of non-steroidal anti-inflammatory drugs (NSAIDs) available in the market. In fact, Meloxicam and Piroxicam, the members of oxicam family are used as an analgesic and anti-inflammatory drugs. Azoles are potential chemical entities with a wide range of pharmacological activities such as anticancer, antibacterial, antifungal, antiviral and antiparasitic. In continuation of our efforts to develop potential multifunctional heterocycles, we have synthesized novel azolylbenzothiazine carboxamides and studied their cytotoxic and antioxidant activities. The results of these aspects will be discussed. © 2020 Author(s).The authors Venkatapuram Padmavathi and Panga Siva Sankar are grateful to CSIR (Council of Scientific and Industrial Research), New Delhi for financial assistance under the major research project

Ecological implications of trace metals in seaweeds: Bio-indication potential for metal contamination in Wandoor, South Andaman Island

AbstractConcentration levels of Mn, Pb, Zn, Cd, Cu and Cr in six seaweed samples (Acetabularia calyculus, Corallina sp., Galaxura marginata, Sargassum duplicatum, Sargassum swartzi and Dictyota bartayresiana) were determined from Wandoor, south Andaman Island. Metals were extracted from sample homogenates and quantified by atomic absorption spectrometry. In the present investigation, heavy metal concentration levels in the following order: Mn>Pb>Cd. It is observed that Zn was only present in free floating brown seaweed S. swartzi. Cu and Cr did not show its presence in any of the seaweeds that was sampled. Metal pollution index (MPI) of six seaweed species were observed in the following decending order: A. calyculus>Corallina sp.>D. bartayresiana>G. marginata>S. duplicatum>S. swartzi. Results showed that chlorophyta, A. calyculus contained the highest concentration of heavy metals as compared to other algal species. One-way analysis of variance (ANOVA) showed that the concentration of metals was significantly different (p<0.05) with respect to different species around the study area

Comparative Analysis of QoS-Aware Routing Protocols for Wireless Sensor Networks

The main ability of wireless sensor networks (WSNs) is communicating and sensing between nodes, which are deployed in a wide area with a large number of nodes. Wireless sensor networks are composed of a large number of sensor nodes with limited energy resources. One critical issue in wireless sensor networks is how to gather sensed information in an energy efficient way, since their energy is limited. The limiting factors of the sensor nodes, such as their finite energy supplies and their moderate processing abilities, as well as the unreliable wireless medium restrict the performance of wireless sensor networks While contemporary best-effort routing approaches address unconstrained traffic, QoS routing is usually performed through resource reservation in a connection-oriented communication in order to meet the QoS requirements for each individual connection. This article surveys a sample of existing QoS-Aware Routing Protocols for Wireless Sensor Networks and highlights their key features, including merits and limitations. Keywords: Wireless sensor networks, Routing protocols, QoS-Aware Routing Protocols