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The recombinant caffeic acid 3-O-methyltransferase gene has been cloned and characterized from Neem. The gene is involved in ferulic acid biosynthesis, a key intermediate component of lignin biosynthesis. Azadirachta indica (Neem) is a highly reputed traditional medicinal plant and is phytochemically well-known for its limonoids. Besides limonoids, phenolics are also distinctively present, which add more medicinal attributes to Neem. Caffeic acid is one of such phenolic compound and it can be converted enzymatically into another bioactive phytomolecule, ferulic acid. This conversion requires transfer of a methyl group from a donor to caffeic acid under the catalytic action of an appropriate methyltransferase. In this study, caffeic acid 3-O-methyltransferase gene from Neem (NCOMT) fruits has been isolated and heterologously expressed in E. coli. The recombinant NCOMT enzyme was purified, which exhibited efficient catalytic conversion of caffeic acid into ferulic acid, a highly potential pharmaceutical compound. The purified recombinant enzyme was physico-kinetically characterized for its catalysis. The analysis of tissue-wide expression of NCOMT gene revealed interesting pattern of transcript abundance reflecting its role in the development of fruit tissues. Further, NCOMT was heterologously overexpressed in Withania somnifera and Ocimum species, to analyze its role in ferulic acid biosynthesis in planta. Thus, the study provides insight for the endogenous role of NCOMT in ferulic acid biosynthesis en route to lignin, an important structural component. To the best of our knowledge, NCOMT pertains to be the first enzyme of the secondary metabolism that has been purified and kinetically characterized from Neem. This study may also have important prospects of applications as the observation on heterologous expression of NCOMT showed its involvement in the maintenance of the in vivo pool of ferulic acid in the plants. Thus, the study involving NCOMT opens up new dimensions of metabolic engineering approaches for the biosynthesis of potential therapeutically important phytomolecules in heterologous systems.

The recombinant caffeic acid 3-O-methyltransferase gene has been cloned and characterized from Neem. The gene is involved in ferulic acid biosynthesis, a key intermediate component of lignin biosynthesis. Azadirachta indica (Neem) is a highly reputed traditional medicinal plant and is phytochemically well-known for its limonoids. Besides limonoids, phenolics are also distinctively present, which add more medicinal attributes to Neem. Caffeic acid is one of such phenolic compound and it can be converted enzymatically into another bioactive phytomolecule, ferulic acid. This conversion requires transfer of a methyl group from a donor to caffeic acid under the catalytic action of an appropriate methyltransferase. In this study, caffeic acid 3-O-methyltransferase gene from Neem (NCOMT) fruits has been isolated and heterologously expressed in E. coli. The recombinant NCOMT enzyme was purified, which exhibited efficient catalytic conversion of caffeic acid into ferulic acid, a highly potential pharmaceutical compound. The purified recombinant enzyme was physico-kinetically characterized for its catalysis. The analysis of tissue-wide expression of NCOMT gene revealed interesting pattern of transcript abundance reflecting its role in the development of fruit tissues. Further, NCOMT was heterologously overexpressed in Withania somnifera and Ocimum species, to analyze its role in ferulic acid biosynthesis in planta. Thus, the study provides insight for the endogenous role of NCOMT in ferulic acid biosynthesis en route to lignin, an important structural component. To the best of our knowledge, NCOMT pertains to be the first enzyme of the secondary metabolism that has been purified and kinetically characterized from Neem. This study may also have important prospects of applications as the observation on heterologous expression of NCOMT showed its involvement in the maintenance of the in vivo pool of ferulic acid in the plants. Thus, the study involving NCOMT opens up new dimensions of metabolic engineering approaches for the biosynthesis of potential therapeutically important phytomolecules in heterologous systems.

Nowadays the number of cars on road are increasing which leads to various problems of which one of them is parking. The public faces various problem such that parking has vacant slot or not. By using the proposed system one can preemptively know whether there is an empty slot or not by looking the LCD display outside the premises as well as android application which will show all the vacant parking slots. The system uses IR sensors alongside engines, LCD and microcontroller for controlling the system. The lcd shows void spaces to new car at gate of parking. On the off chance that no parking spot is accessible the system does not open the door and lcd says parking full.If space is vacant system enables car to enter the parking and shows void openings where client can stop. To identify vehicle opening inhabitance the system utilizes IR sensors. Additionally System utilizes IR sensors to distinguish vehicles touching base at stopping gates,to open the doors naturally on vehicle landing. The microcontroller is utilized to encourage the working of the whole system. Android application can also be used to find alternate parking premises in the area by displaying number of vacant parking slots in them too. It also allow user to book parking slot 15 minutes before user arrives else it will get cancelled if the user doesn’t arrive in the specified time.

The biosynthetic potential of life forms can be reinvigorated by genome manipulation and construction of genetic circuits and complex regulatory networks to program new functions in the platform organisms. This requires in-depth understanding of the integrative complexity of biological design principles and applications of different foundational scientific fields with engineering principles; this interdisciplinary approach of biology and engineering is called as synthetic biology. In recent years, fabrication of biological toolkits has expanded from constructing novel genetic parts to creating synthetic life. The construction of new biological parts or engineering of designer cells with precise control of cellular programs or redesigning of natural biological systems has been proved very beneficial in different agri-food and medical applications. This chapter discusses the basics of synthetic biology, the relevant molecular tools, and root systems in synthetic biology.

Establishment of an efficient in vitro regeneration system is a pre-requisite for genetic modification of plants. The present study reports a high efficiency regeneration protocol for cauliflower cv. Pusa Meghna using cotyledon and hypocotyl explants cultured on different growth media with varying combinations of plant growth regulators. Maximum shoot regeneration response from hypocotyl (95.76%) and cotyledon (83.01%) was obtained on MS medium containing 1mg/L BAP and 1 mg/L BAP+0.1 mg/L NAA+0.5 mg/L AgNO3+0.1 mg/L GA3, respectively. Further optimization of rooting media for development of full grown plants showed maximum root regeneration of 92.20% and 94.86% from hypocotyl and cotyledon explants grown on MS medium containing 0.5 mg/L IBA and 0.2 mg/L IBA, respectively. Utilization of the present established regeneration protocol would serve as a valuable tool for various crop improvement studies in cauliflower.

Ocimum species commonly referred to as "Tulsi" are well-known for their distinct medicinal and aromatic properties. The characteristic aroma of Ocimum species and cultivars is attributed to their specific combination of volatile phytochemicals mainly belonging to terpenoid and/or phenylpropanoid classes in their essential oils. The essential oil constituents are synthesized and sequestered in specialized epidermal secretory structures called as glandular trichomes. In this comparative study, inter- and intra-species diversity in structural attributes and profiles of expression of selected genes related to terpenoid and phenylpropanoid biosynthetic pathways have been investigated. This is performed to seek relationship of variations in the yield and phytochemical composition of the essential oils. Microscopic analysis of trichomes of O. basilicum, O. gratissimum, O. kilimandscharicum, and O. tenuiflorum (green and purple cultivars) revealed substantial variations in density, size, and relative proportions of peltate and capitate trichomes among them. The essential oil yield has been observed to be controlled by the population, dominance, and size of peltate and capitate glandular trichomes. The essential oil sequestration in leaf is controlled by the dominance of peltate glandular trichome size over its number and is also affected by the capitate glandular trichome size/number with variations in leaf area albeit at lower proportions. Comprehension and comparison of results of GC-MS analysis of essential oils showed that most of the Ocimum (O. basilicum, O. tenuiflorum, and O. gratissimum) species produce phenylpropanoids (eugenol, methyl chavicol) as major volatiles except O. kilimandscharicum, which is discrete in being monoterpenoid-rich species. Among the phenylpropanoid-enriched Ocimum (O. basilicum, O. gratissimum, O. tenuiflorum purple, O. tenuiflorum green) as well, terpenoids were important constituents in imparting characteristic aroma. Further, comparative abundance of transcripts of key genes of phenylpropanoid (PAL, C4H, 4CL, CAD, COMT, and ES) and terpenoid (DXS and HMGR) biosynthetic pathways was evaluated vis-a-vis volatile oil constituents. Transcript abundance demonstrated that richness of their essential oils with specific constituent(s) of a chemical group/subgroup was manifested by the predominant upregulation of phenylpropanoid/terpenoid pathway genes. The study provides trichomes as well as biosynthetic pathway-based knowledge for genetic improvement in Ocimum species for essential oil yield and quality.

Blockchain is another approach to store and record exchanges. It is particularly similar to a conventional database but the blocks are connected together cryptographically with a specific end goal to ensure that they are sealed. Blockchain technology was developed in 2008 to make digital money. The essential case that individuals know about is most likely a BITCOIN. Individuals use it as a digital money, however individuals have begun to use it more for things like their previous testament, endorsement and declare that the valuable stone is not a blood jewel, rather anyhow not to share information. Since, there is no regulatory framework in transactions between two or more parties but there is a backend activity (government) which make sure of all the transactions made by a person or a buyer who owns large number of cryptocurrencies.

There are millions of data been generated in various forms around us regularly which is approximately 2.5 quintillion bytes on the daily basis and most of the data are coming from social sites. Social sites are one of the biggest contributor of big data generation. Big data is a technology which deals with extremely large data which is used to analyze different patterns, associations, trends and largely in human behavior and interactions computationally where social media data came in use. As sharing information as per one's daily basis activities and many other forms online is the main objective of social media whereas, users are unable to understand the process and they keep on uploading their personal data without having the idea that all this is contributing big data and is continually collecting at various platforms which might affect users privacy. In this paper, here analyzed big data privacy in social media and some major issues related its privacy and heavy rate of growing data per day.

Ocimum kilimandscharicum is unique in possessing terpenoids whereas other Ocimum species are renowned for phenylpropanoids as major constituents of essential oil. The key enzyme of MVA/terpenoid metabolic pathway viz 3-hydroxy-3-methylglutaryl Co-A reductase (OkHMGR) of 1.7-Kb ORF encoding ~60-kDa protein was cloned from O. kilimandscharicum and its kinetic characteristics revealed the availability of HMG-CoA as a control point of MVA-pathway. Transcript profiling of the OkHMGR elucidated tissue-specific functions of the gene in flower and leaf tissues in accumulation of terpenoidal essential oil. OkHMGR was differentially regulated in response to exposure to methyl-jasmonate, salicylic-acid, and stress conditions such-as salt and temperature stress, demonstrating its key role in managing signaling and stress-responses. To elucidate its functional role, OkHMGR was transiently over-expressed in homologous and heterologous plants such as O. sanctum, O. basilicum, O. gratissimum, Withania somnifera and Artemisia annua. The over-expression and inhibition dual strategy revealed that the additional OkHMGR in-planta could afford endogenous flow of isoprenoid units towards synthesis of terpenoids. The present study provides in-depth insight of OkHMGR in regulation of biosynthesis of non-plastidal isoprenoids. This is first report on any gene of MVA/isoprenoid pathway from under-explored Camphor Tulsi belonging to genus Ocimum. Studies also suggested that OkHMGR could be a potential tool for attempting metabolic engineering for enhancing medicinally important terpenoidal metabolites in plants.

Withania somnifera one of the most reputed Indian medicinal plant has been extensively used in traditional and modern medicines as active constituents. A high frequency genotype and chemotype independent Agrobacterium-mediated transformation protocol has been developed for W. somnifera by optimizing several factors which influence T-DNA delivery. Leaf and node explants of Withania chemotype was transformed with A. tumefaciens strain GV3101 harboring pIG121Hm plasmid containing the gusA gene encoding β-glucuronidase (GUS) as a reporter gene and the hptII and the nptII gene as selection markers. Various factors affecting transformation efficiency were optimized; as 2 days preconditioning of explants on MS basal supplemented with TDZ 1 μM, Agrobacterium density at OD600 0.4 with inclusion of 100 μM acetosyringone (As) for 20 min co-inoculation duration with 48 h of co-cultivation period at 22 °C using node explants was found optimal to improved the number of GUS foci per responding explant from 36 ± 13.2 to 277.6 ± 22.0, as determined by histochemical GUS assay. The PCR and Southern blot results showed the genomic integration of transgene in Withania genome. On average basis 11 T0 transgenic plants were generated from 100 co-cultivated node explants, representing 10.6 % transformation frequency. Our results demonstrate high frequency, efficient and rapid transformation system for further genetic manipulation in Withania for producing engineered transgenic Withania shoots within very short duration of 3 months.

System based distributed computing is quickly growing as a contrasting option to conventional office based figuring. As distributed computing turns out to be more far reaching, the vitality utilization of the system and processing assets for the cloud will develop. It is going on during an era when there is expanding consideration being paid to the need to oversee energy utilization in Data and Correspondence Innovation (ICT) area. In this paper, I have introduced an investigation of vitality utilization and stage free administration (PaaS). PaaS give offers by and large the stage administrations for application advancement. The stage encourages the development of uses which guarantees framework vitality productivity. It is a product vitality proficiency which measures the vitality required to perform sorting calculation that can deal with huge measure of information. Distributed computing can empower more vitality productive utilization of figuring force. Under a few circumstances distributed computing can expend more vitality than conventional registering where every client plays out all figuring all alone PC.

In this paper, a 3-D crossbar switch is designed for 3-D mesh Network-on-Chip to meet the current requirements of smaller area, power efficiency and high speed. To increase the overall data transfer rate of the network, the number of ports are added in 3-D crossbar switch and found that it has been working at the same power as the 2-D crossbar switch (with lesser ports). The functional verification of the crossbar switch design has been done on ModelSim6.4a. Xilinx13.3 has been used for synthesis. Also, FPGA verification of the proposed design has been done.

Network-on-Chip is an emerging paradigm for integrating very high number of Intellectual Property blocks on a single Integrated Chip. In this paper, crossbar switch is designed for 2-D Mesh Network-on-Chip to meet the current requirements of high speed networks. The design reduced the power consumption significantly and can meet the scalable bandwidth. The functional verification of the crossbar switch design using ModelSim 6.4a has been shown. Synthesis has been performed on Xilinx 13.3 and it is verified using FPGA.

Rose-scented geranium (Pelargonium spp.) is one of the most important aromatic plants and is well known for its diverse perfumery uses. Its economic importance is due to presence of fragrance rich essential oil in its foliage. The essential oil is a mixture of various volatile phytochemicals which are mainly terpenes (isoprenoids) in nature. In this study, on the geranium foliage genes related to isoprenoid biosynthesis (DXS, DXR and HMGR) were isolated, cloned and confirmed by sequencing. Further, the first gene of 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway, 1-deoxy-d-xylulose-5-phosphate synthase (GrDXS), was made full length by using rapid amplification of cDNA ends strategy. GrDXS contained a 2157 bp open reading frame that encoded a polypeptide of 792 amino acids having calculated molecular weight 77.5 kDa. This study is first report on heterologous expression and kinetic characterization of any gene from this economically important plant. Expression analysis of these genes was performed in different tissues as well as at different developmental stages of leaves. In response to external elicitors, such as methyl jasmonate, salicylic acid, light and wounding, all the three genes showed differential expression profiles. Further GrDXS was over expressed in the homologous (rose-scented geranium) as well as in heterologous (Withania somnifera) plant systems through genetic transformation approach. The over-expression of GrDXS led to enhanced secondary metabolites production (i.e. essential oil in rose-scented geranium and withanolides in W. somnifera). To the best of our knowledge, this is the first report showing the expression profile of the three genes related to isoprenoid biosynthesis pathways operated in rose-scented geranium as well as functional characterization study of any gene from rose-scented geranium through a genetic transformation system.

Withania somnifera Dunal, is one of the most commonly used medicinal plant in Ayurvedic and indigenous medicine traditionally owing to its therapeutic potential, because of major chemical constituents, withanolides. Withanolide biosynthesis requires the activities of several enzymes in vivo. Cycloartenol synthase (CAS) is an important enzyme in the withanolide biosynthetic pathway, catalyzing cyclization of 2, 3 oxidosqualene into cycloartenol. In the present study, we have cloned full-length WsCAS from Withania somnifera by homology-based PCR method. For gene function investigation, we constructed three RNAi gene-silencing constructs in backbone of RNAi vector pGSA and a full-length over-expression construct. These constructs were transformed in Agrobacterium strain GV3101 for plant transformation in W. somnifera. Molecular and metabolite analysis was performed in putative Withania transformants. The PCR and Southern blot results showed the genomic integration of these RNAi and overexpression construct(s) in Withania genome. The qRT-PCR analysis showed that the expression of WsCAS gene was considerably downregulated in stable transgenic silenced Withania lines compared with the non-transformed control and HPLC analysis showed that withanolide content was greatly reduced in silenced lines. Transgenic plants over expressing CAS gene displayed enhanced level of CAS transcript and withanolide content compared to non-transformed controls. This work is the first full proof report of functional validation of any metabolic pathway gene in W. somnifera at whole plant level as per our knowledge and it will be further useful to understand the regulatory role of different genes involved in the biosynthesis of withanolides.

An efficient and reproducible Agrobacterium-mediated genetic transformation of Withania coagulans was achieved using leaf explants of in vitro multiple shoot culture. The Agrobacterium strain LBA4404 harboring the binary vector pIG121Hm containing β-glucuronidase gene (gusA) under the control of CaMV35S promoter was used in the development of transformation protocol. The optimal conditions for the Agrobacterium-mediated transformation of W. coagulans were found to be the co-cultivation of leaf explants for 20 min to agrobacterial inoculum (O.D. 0.4) followed by 3 days of co-cultivation on medium supplemented with 100 μM acetosyringone. Shoot bud induction as well as differentiation occurred on Murashige and Skoog medium supplemented with 10.0 μM 6-benzylaminopurine, 8.0 μM indole 3-acetic acid, and 50.0 mgl(-1) kanamycin after three consecutive cycles of selection. Elongated shoots were rooted using a two-step procedure involving root induction in a medium containing 2.5 μM indole 3-butyric acid for 1 week and then transferred to hormone free one-half MS basal for 2 weeks. We were successful in achieving 100 % frequency of transient GUS expression with 5 % stable transformation efficiency using optimized conditions. PCR analysis of T0 transgenic plants showed the presence of gusA and nptII genes confirming the transgenic event. Histochemical GUS expression was observed in the putative transgenic W. coagulans plants. Thin layer chromatography showed the presence of similar type of withanolides in the transgenic and non-transgenic regenerated plants. A. tumefaciens mediated transformation system via leaf explants developed in this study will be useful for pathway manipulation using metabolic engineering for bioactive withanolides in W. coagulans, an important medicinal plant.

E-Advertisements have made possible to allow marketers for approaching target segments in the most measurable, interactive and more essentially, cost-effective ways. However, Neural Network is a forecasting tool for dynamic and changing market environments. A Strong advantage of neural networks is that a properly trained network can be considered experts with regard to the particular output project for which it was designed to examine. This paper gives brief view about various e-advertisement Payment trends. Various sector wise e-advertisement related data from 2008 to 2013 have been collected from IAB (Internet Advertisement Bureau) and applied the Back Propagation technique of Neural Network for predicting ratio of cost models in E-advertisements. Effective use of data mining will ear mark of E-advertisement in various industries like consumer service, retail, auto, travel, computing, media, financial service, telecommunication etc.

Clustering is one of the data analysis methods that are widely used in data mining. In this method, we partitioned the data into different subset which is known as cluster. Cluster analysis is the data reduction toll for classifying a “mountain‟ of information into manageable meaningful piles. This method is vast research area in the field of data mining. In this paper, a partitioning clustering method that is K-Medoids algorithm is used with Bat algorithm. We proposed a new algorithm based on the echolocation behaviour of bats to know the initial value to overcome the K-Medoids issues. In this algorithm, we can find the initial representative object easily with the help of using Bat algorithm. They provide us better cluster analysis and we can achieve efficiency. This paper introduces the combination of K-Medoids clustering algorithm and Bat Algorithm. In this paper we show the difference between K-Medoid Clustering Technique with Bat Algorithm & K-medoid itself.

Secondary metabolites, the biological compounds secreted by plants as an aid to support their growth and development under stress conditions or as a part of their defense mechanism, now hold equal importance for mankind who employs it immensely for medication, flavorings, aroma, etc. Wide applicability of these compounds instigates one to understand the biosynthesis, structure and regulation of these bioactive molecules. Terpenoids form the largest group of secondary metabolites which comprise of a wide range of structurally and functionally distinct metabolites synthesized either via mevalonate pathway or non-mevalonate pathway. Targeting a key regulatory enzyme of this pathway, modulation of which would alter the carbon flux would be beneficial to enhance our knowledge about the above issue. For this the transcriptome (from SRA) of different Ocimum species was mined out for important pathway genes using various bioinformatics approaches. Amongst them 3-hydoxy 3-methyl glutaryl CoA reductase (HMGR) was selected which is the rate limiting enzyme in mevalonate pathway which controls the conversion of HMG-CoA to mevalonic acid. Isolation, cloning, protein expression, purification, etc. would be discussed in detail in the meeting. Full length protein was also characterized through bioinformatics tools to study its structure, properties, conserved domains, etc. Increase in secondary metabolite production by alteration of HMGR pool along with transcript modulation studies in planta revealed that HMGR gene governs the biosynthesis of secondary metabolites. Transcriptome mapping of different HMGR homologs which on comparison within member of same genus revealed its divergent nature which could account to its multifunctional role in different plants. Besides, providing a deep insight about the enzyme function combination of such molecular, transgenic and bioinformatics tools would help to develop strategies to engineer the HMGR mediated flux and also valuable metabolites in plants.

Withania somnifera Dunal is one of the most commonly used plants in Ayurvedic and indigenous system of medicine in India for over thousands of years. In view of its varied therapeutic potential, the plant has also been the subject of considerable scientific attention. The major chemical constituents of the Withania genus, the withanolides, are a group of naturally occurring C28-steroidal lactones built on an intact or rearranged ergostane framework, in which C22 and C26 are oxidized to form a six-member lactone ring. In recent years, numerous pharmacological investigations have been carried out utilizing W. somnifera extracts and several patents have been filed on pharmacological and medicinal importance of withanolides and extracts of W. somnifera, individually or in combination. Considering the immense importance of withanolides for medicinal purposes, the establishment of strategies to improve withanolides yield are highly desirable. Under natural conditions, W. somnifera possesses restricted levels of withanolides then, alternatives for obtaining withanolides in better yields are imperative. In vitro approaches followed by metabolic engineering could be attractive tools to achieve this goal. Therefore, we present here an overview of the development of various protocols for in vitro tissue regeneration from W. somnifera and in vitro secondary metabolite production as well. The review also gives an account of selected patents on various important activities of phytochemicals and extracts of W. somnifera.