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Education is a pillar for success and every teacher tries to make sure to provide a better way for a student to learn. As time passed, the way of teaching also changed. Starting from no visual to chart paper to images to slides or real objects, the way of teaching changed to enhance the student's experience in learning. Now we are at the stage where slides/chart papers are getting old and 3D/4D views are coming into the picture. New technology like Metaverse is going to change interactive learning. An immersive experience will be new for teachers as well as students.

Our earlier studies have established improved cold tolerance in the transgenic tomato (Solanum lycopersicum cv. Pusa Ruby) developed through constitutive over expression of Nicotiana tabacum Osmotin gene. To understand the molecular responses, micro-array analysis was performed in the transgenic and the wild type on cold (4.0 °C) treatment for 2 and 24 h using Affymetrix GeneChip® Tomato Genome Array. The principal component analysis plots placed the wild type and transgenic in different regions, indicating differential transcript regulation on cold exposure. The differential gene expression analysis performed using pairwise comparison for transgenic and wild type indicated modulation of 307 out of 3770 genes, by 10 or higher fold on cold treatment. Sugar and amino acid metabolism together constituted the maximum of the annotated products of the differentially expressed genes. The results revealed modulation of expression of responsive genes involved in various functions in the transgenic tomato on cold treatment.

Timur (Zanthoxylum armatum DC.) also known as toothache tree or winged prickly ash is an important large thorny wild shrub of Rutaceae family. Owing to its high medicinal properties, the plant population is rapidly depleting due to the unsustainable premature harvesting from open-access forests. Because of which the plant species can become endangered or rare in the absence of conservation measures through its mass propagation. Seed propagation in Timur is a time consuming and tedious process due to the requirement for stratification. Here, we report the effect of different seed pre-treatments including water soaking, potassium nitrate (300 mM), sodium chloride (75 mM) and concentrated sulfuric acid treatments followed by water soaking on seed germination in Timur. Seed pre-treatment with acid for 120 min resulted in rapid and synchronous seed germination whereas the control, water-soaked, potassium nitrate or sodium chloride-treated seeds failed to germinate during the observation period of 30 days. The plantlets from acid-treated seeds further established successfully. Thus, the results of the present study may be applicable in large-scale simple and low-cost mass propagation of the plant as a source of medicines and other purposes.

According to world health statistics 285 million out of 7.6 billion population suffers visual impairment; hence 4 out of 100 people are blind. Absence of vision restricts the mobility of a person to pronounced extent and hence there is a need to build an explicit device to conquer guiding aid to the prospect. This paper proposes to build a prototype that performs real time object detection using image segmentation and deep neural network. Further the object, its position with respect to the person and accuracy of detection is prompted through speech stimulus to the blind person. The accuracy of detection is also prompted to the device holder. This work uses a combination of single-shot multibox detection framework with mobileNet architecture to build rapid real time multi object detection for a compact, portable and minimal response time device construction.

Defence sector desires to attain energy self-sufficiency and security. In recent years, emergence of biofuel as an alternative source has raised the hopes of Defence. Ethanol and bio-diesel are currently being used as blends in different parts of the world. While, bio-diesel is mostly being blended in 2-20% in different parts of the world, ethanol blending has reached upto 85%. Owing to the sustainability reasons, the choice of feedstock for ethanol production is gradually changing from corn to lignocelluloses biomass. Jatropha curcas, is still the choice feedstockfor bio-diesel in most third world countries. This institute has put in rigrous efforts to identify high yielding varieties of Jatropha, improving its yield, standardizing trans-esterification to obtain high quality bio-diesel and its trials and testing in various vehicles and equipment. Second generation biofuels using biomass such as farm and forest wastes as feedstocks are promising in terms of their overall sustainability and volume produced. They can be used as drop in fuels. However, time is required to utilize their potential fully. Algae, the third generation biofuel feedstock still needs extensive R&D to make it economically sustainable. Whatever, the technology used, defence forces will accept any biofuel, which should be available constantly and priced below the existing petroleum fuels. The scope of producing by-products and finding a lucarative market for these products can ensure that prices ofbiofuels remain lower than the petroleum fuels

Crop yield is challenged every year worldwide by changing climatic conditions. The forecasted climatic scenario urgently demands stress-tolerant crop varieties to feed the ever-increasing global population. Molecular breeding and genetic engineering approaches have been frequently exploited for developing crops with desired agronomic traits. Recently, microRNAs (miRNAs) have emerged as powerful molecules, which potentially serve as expression markers during stress conditions. The miRNAs are small non-coding endogenous RNAs, usually 20–24 nucleotides long, which mediate post-transcriptional gene silencing and fine-tune the regulation of many abiotic- and biotic-stress responsive genes in plants. The miRNAs usually function by specifically pairing with the target mRNAs, inducing their cleavage or repressing their translation. This review focuses on the exploration of the functional role of miRNAs in regulating plant responses to abiotic and biotic stresses. Moreover, a methodology is also discussed to mine stress-responsive miRNAs from the enormous amount of transcriptome data available in the public domain generated using next-generation sequencing (NGS). Considering the functional role of miRNAs in mediating stress responses, these molecules may be explored as novel targets for engineering stress-tolerant crop varieties.

Here, we report a whole transcriptome analysis of LlaNAC gene (from Lepidium latifolium) containing transgenic tobacco line (NC10) and wild type (WT), to attain deeper knowledge into the downstream genes activated by the over-expressing transgene. Transcriptome sequencing of NC10 and WT samples generated huge data using Illumina platform. The maximum number of unigenes GO annotated were of Biological process (8988, 3209) followed by molecular function (5155, 2577) and cellular components (3826, 1583) for WT and NC10 samples respectively. KEGG Pathway analysis revealed the unigenes were enriched in different functional pathway categories. The unigenes whose products involved in carbohydrate metabolism, glycan metabolism, and secondary metabolites synthesis were more for NC10 library in comparison to WT. Greater variety of transcription factors were involved in transgenic than wild-type plants. Genes like, Copia-like retrotransposable element, Peroxidase 64-like, Peptidyl-prolyl cis-trans isomerise, Cytochrome P450, Lipoyl synthase, CBL-interacting serine/threonine-protein kinase 5-like etc. were found differentially expressed in both the samples. Promoter analysis of these differentially expressed genes have elements for defence and stress response, abscisic acid response, shoot specific expression and light response, etc. In summary this study reports the involvement of the overexpressed genes in the dual action of cold tolerance and biomass accumulation, as sugars participate in both of these activities of the cell.

Our earlier study has identified Lepidium latifolium Ran (LlaRan) as a cold stress downregulated gene from L. latifolium, a cold desert plant. Further, independent transgenic tobacco (Nicotiana tabacum L. var Xanthi) lines overexpressing LlaRan were developed for functional characterization of the gene. We analysed the tolerance of tobacco plants over expressing LlaRan gene to salt and PEG stresses at germination which represents a critical stage in life cycle of the plants. Germination performance of the four independent transgenic lines (T1 generation) were assessed for germination under distilled water, NaCl (150 mM) and osmotic (15% PEG 4000) stresses. The emergence of radical, plumule and cotyledonary leaf were recorded for 10 days on daily basis and its rate assessed in terms of Timson’s index. It was observed that the germination of wild type tobacco seeds was better than the assessed transgenic lines. It may be inferred that the transgenic lines seeds under study were more sensitive to salt and drought stress elucidating their hypersensitivity at germination level.

Four tobacco lines over-expressing LlaNAC (FJ423495) gene have been assessed for various parameters like abiotic stress response, phytochemical constituents, biomass composition, response to elevated carbon dioxide (CO2), etc. All the analyses have been carried out in T2 generation of the transgenic plants with proven genetic stability. In response to the heat and drought stress, the accumulation of antioxidant enzymes elevated up to 2.5-fold. Photosynthetic pigments, i.e., Chl a, Chl b and xanthophylls, and phenolics in the transgenic lines were found significantly (two–fourfold) enhanced in comparison to wild-type (WT) plants. The transgenic lines were also shown capturing 5–10 times more CO2 from the environment, which presumably gets accumulated as biomass. However, ratio of dry weight to fresh weight was slightly in favor of the WTs. Cellulose, hemi-cellulose and lignin contents in stem and leaves of each of the transgenic lines varied, and overlapped with the lignocellulosic contents of the WT plants. In conclusion, LlaNAC gene shows promise for exploitation in design of the future crops with various end uses.

Jatropha curcas (Jatropha), one of the most popular biofuel crops, is also one of the most controversial crops. It is generally hailed as biofuel mandate crop in Asia and Africa, but its low genetic diversity in these continents has failed most of the crop improvement programs initiated so far. Breeding through the utilization of germplasm from Mexico is one of the priority areas for its genetic improvement. Nevertheless, proper agricultural practices, which may vary from region to region, are also important, not only to obtain optimum yields from improved germplasm but also the best yield from the germplasm introduced in different agroecological systems from the wild around the Jatropha belt. It is desired that Jatropha plantations sustain themselves, and resources meant for agriculture for food shall not be diverted towards agriculture for fuels. Irrigation, nevertheless, plays an important role in ensuring economically beneficial yields from jatropha cultivation. Similarly, application of fertilizers (NPK) is important for obtaining a good harvest from Jatropha fields. While, biodiesel, obtained through transesterification, is the major fuel obtained from Jatropha, many other fuels like biogas, fuel briquettes, Fischer-Tropsch (FT) diesel, ethanol, etc. can also be obtained as by-products. Utilization of biodiesel blended with fossil diesel is least technologically challenging with regard to Jatropha-based economy. However, availability of desired quantities of fuel is a challenge. Despite all pros and cons associated with Jatropha, it is still considered as an ideal feedstock for biodiesel, which doesn’t compete with food crops, and returns are offered through several by-products.

There is much debate about the adequacy of the U.S. physician workforce and projections of its future size, distribution and composition. Beginning with 3 observations about the workforce we believe are largely not subject to dispute, we address the debate by providing an overview of the current state of the workforce and Graduate Medical Education in the United States; a brief history of both calls for graduate medical education reform since 1910 and the recent, intense debate about the reliability of workforce projections; and a discussion of the challenges to understanding the physician workforce. We draw 3 concluding observations: (1) Precisely because projections can be unpredictable in their impact on both physician workforce behavior and public policy development, policy makers need to devote more attention to workforce projections, not less. (2) More research devoted specifically to the workforce implications of delivery and payment reforms is strongly needed. (3) Such research must be pursued with a sense of urgency, given the rapid aging of the Baby Boom generation, which will put a disproportionate demand on the nation's physician workforce.

The ability of Camelina sativa to withstand salinity stress in vitro by adding NaCl (0, 25, 50, 75, 100, 125, 150, 175, 200mM) in Murashige and Skoog basal medium was studied. Performance of the plants was measured in terms of various growth parameters and physiological and biochemical tests performed on fully grown plants. The germination capacity, cotyledon unfolding and first true leaf emergence was reduced by 30.6, 17.3, and 28.8%, respectively in 200mM salt treatment with respect to control. The plant height, relative water content, and plant water content were decreased by 85.4, 10.8, and 9.8%, respectively, in stressed plants with respect to control. A decrease in chlorophyll a and b and total chlorophyll contents (by 81.3%), as well as of protein content was registered. Electrical conductivity increased by 52.8% in stressed plants over control, as expected. Other stress indicators like guiacol peroxidase activity and malondialdehyde also increased with respect to control. At salt concentrations lower than 200mM, no clear cut retardation effects were seen. Thus, the present study opens up the scope of further assessment of survivability of camelina in salt contaminated soils.

The present study reports engineered cold tolerance and toxicity analysis in genetically modified tomato (Solanum lycopersicum L. cv. Pusa Ruby) developed through constitutive over expression of Nicotiana tabacum Osmotin gene. Rate of seed germination, seedling establishment and growth remained unaffected in the transgenic tomato in response to a low temperature (15 °C) treatment, but were significantly (P ≤ 0.05) reduced in the wild type. At reproductive stage, the wild type plants failed to recover at the low temperature (4.0 °C) treatment for 10 days but the transgenic plants survived successfully without any leaf senescence or other visible chilling injury symptoms. The quantitative transcript expression analysis confirmed up regulation of the transgene by 55% in the transgenic plants on cold treatment for 2 h whereas, the transcripts were not detected in the wild type. Containment evaluation under normal environmental conditions revealed similar morphology in both the transgenic and wild type tomato plants however an average fruit yield was higher in the transgenic plants (725.91 ± 39.27 g) than the wild type (679.84 ± 28.80 g). The composition of mature fruits in terms of element content was at par in both the transgenic and wild type except significantly higher Ca and Mg contents in the transgenic fruits than that of the wild type. Further, acute and sub-acute toxicity tests conducted in the adult female Wister rats revealed no mortality or significant changes in general and psychological behaviour, at par food intake and body weight and, normal biochemical, and hematological parameters for animals fed with the wild type or transgenic tomato fruits as compared to the control group, confirming its safety for animal consumption.

The changes to Medicare payment documentation guidelines proposed by the Centers for Medicare & Medicaid Services may simplify documentation requirements. However, this commentary argues that the p...

The present study reports engineered cold tolerance and toxicity analysis in genetically modified tomato (Solanum lycopersicum L. cv. Pusa Ruby) developed through constitutive over expression of Nicotiana tabacum Osmotin gene. Rate of seed germination, seedling establishment and growth remained unaffected in the transgenic tomato in response to a low temperature (15 °C) treatment, but were significantly (P ≤ 0.05) reduced in the wild type. At reproductive stage, the wild type plants failed to recover at the low temperature (4.0 °C) treatment for 10 days but the transgenic plants survived successfully without any leaf senescence or other visible chilling injury symptoms. The quantitative transcript expression analysis confirmed up regulation of the transgene by 55% in the transgenic plants on cold treatment for 2 h whereas, the transcripts were not detected in the wild type. Containment evaluation under normal environmental conditions revealed similar morphology in both the transgenic and wild type tomato plants however an average fruit yield was higher in the transgenic plants (725.91 ± 39.27 g) than the wild type (679.84 ± 28.80 g). The composition of mature fruits in terms of element content was at par in both the transgenic and wild type except significantly higher Ca and Mg contents in the transgenic fruits than that of the wild type. Further, acute and sub-acute toxicity tests conducted in the adult female Wister rats revealed no mortality or significant changes in general and psychological behaviour, at par food intake and body weight and, normal biochemical, and hematological parameters for animals fed with the wild type or transgenic tomato fruits as compared to the control group, confirming its safety for animal consumption. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-018-1432-7) contains supplementary material, which is available to authorized users.

The fossil fuels, besides being polluting energy sources, are depleting at faster rate with the economic development. Therefore, development of an alternate renewable and clean energy source has gained serious attention worldwide. According to the feedstock and technology used, the biofuel technologies have been categorized as the first-, second-, third-, and fourth-generation biofuels. The traditional first-generation biofuels were derived from vegetable oil and other crops. However, they have serious economic and environmental constraints attached besides putting up competition to agri-food production. In contrast, second- and third-generation biofuels derived from biomass and algae, respectively, are far more energy efficient and sustainable at the ecological and societal level. Furthermore, omics approaches are being used for developing the fourth-generation biofuels from engineered organisms and are believed to be a potential renewable, cost effective, and cleaner energy source to meet the global energy demand. This chapter describes the recent technological developments for making the biofuel production economically competitive and environment friendly.

Molecular markers, due to their stability, cost-effectiveness and ease of use provide an immensely popular tool for a variety of applications including genome mapping, gene tagging, genetic diversity diversity, phylogenetic analysis and forensic investigations. In the last three decades, a number of molecular marker techniques have been developed and exploited worldwide in different systems. However, only a handful of these techniques, namely RFLPs, RAPDs, AFLPs, ISSRs, SSRs and SNPs have received global acceptance. A recent revolution in DNA sequencing techniques has taken the discovery and application of molecular markers to high-throughput and ultrahigh-throughput levels. Although, the choice of marker will obviously depend on the targeted use, microsatellites, SNPs and genotyping by sequencing (GBS) largely fulfill most of the user requirements. Further, modern transcriptomic and functional markers will lead the ventures onto high-density genetic map construction, identification of QTLs, breeding and conservation strategies in times to come in combination with other high throughput techniques. This review presents an overview of different marker technologies and their variants with a comparative account of their characteristic features and applications.

Repetitive sequence elements refer to sequences which are repeated a number of times in the genome and are generally composed of either the interspersed elements or tandem repeats, which may be coding or non-coding. Solanum genomes, like that of potato and tomato, are exceptionally rich in repetitive elements and have evolved by the multidirectional activities of the repetitive elements. Interspersed elements, in particular, have contributed to the evolution of Solanum genomes, not only by their amplification in the genome, but also by their association with different genes. The genomic proximity of interspersed elements to genes may influence the expression patterns, development of alternative functions, emergence of new genes or elimination of existing genes. Long terminal repeat (LTR) retrotransposons represent the most abundant class of repetitive elements among all the repeats and they are also considered the progenitors of some genes and other types of repeats such as microsatellites. Centromeric, telomeric, minisatellites and microsatellites constitute tandem repeats, which are abundant in the Solanum genomes. The patterns of occurrence and abundance of repeats vary among different species of Solanum, although a careful analysis reveals a number of perceptible evolutionary trends. Tandem repeats, particularly microsatellites, have frequently been exploited for the development of molecular markers for diverse applications in potato. The present chapter deals with the occurrence and distribution of different categories of repetitive elements in potato and the related Solanum genomes.

We report elevated biomass and altered growth characteristics of tobacco plants up on transformation with a NAC (NAM, ATAF1/2,CUC2) gene (GenBank Accession FJ754254) isolated from Lepidium latifolium L. (LlaNAC). Transgenic plants showed significant differences in fresh weight, midrib length of longest leaf, leaf area, height of the plant, root and shoot weights, etc. during vegetative phase. On 100th day after sowing (DAS), plants of transgenic lines were 2-3 times taller than the wild type plants, though no significant difference was recorded in moisture contents of any of the plant tissues. Over-expression of NAC gene up to 2,000 fold was recorded in leaves of transgenic plants on 100th DAS. Interestingly, transgenic plants showed significantly shortened (P(t) = 0.02-0.04) life cycle, as they showed a completely altered growth behaviour. Transgenic plants entered reproductive phase earlier by 60 days, with lines NC2 and NC7b entering first, followed by line NC10. However, the time period spent in the reproductive phase by the plant was nearly twice in case of transgenic lines NC2, NC7b and NC10, as compared to the wild type plants. Despite that, these lines completed their life cycle in 45-60 days lesser than the time taken by wild-type tobacco plants. No difference was recorded in fruit and seed yield of transgenic or wild type plants. To the best of our knowledge, this is the first report on over-expression of NAC gene causing altered growth and biomass patterns. We expect this study to become an important reference towards future engineering of plants for fuel and fodder purposes.

Lepidium latifolium L., a weed distributed in the Ladakh region of Himalayan range, belongs to Brassicaceae family and reported to withstand low temperature stress

We have isolated and in silico characterized a cold regulated plastocyanin encoding gene from Lepidium latifolium L designated as LlaDRT. Its cDNA sequence (JN214346) consists of a 504 bp ORF, 48 and 205 bp of 5' and 3' UTR regions, respectively encoding a protein of 17.07 KDa and pI 4.95. In silico and phylogenetic analysis of LlaDRT suggested that the protein has features of a typical plastocyanin family member and of a nearest relative of the predominant isoform of Arabidopsis (PETE2) plastocyanin. Validation of stress response of LlaDRT by qPCR under different abiotic stress regulators viz salicylic acid, jasmonic acid, calcium chloride, ethylene and abscisic acid revealed its possible regulation and crosstalk amongst different pathways.

The present study reports a simple, efficient and high-throughput method for confirmation of neomycin phosphotransferase transgenic tomato (Solanum lycopersicum L. cv. Pusa Ruby) plants. For confirmation at seedlings and maturity stage, transgenic (T7) and wild type plants were sprayed with various Kanamycin concentrations (100–400 mg/l). Only the wild type plants sprayed with concentrations ≥300 mg/l developed typical chlorotic symptoms after 2 days, thus differentiating from the transgenic. For transgenic confirmation at earlier stage, transgenic and wild type seeds were germinated on filter paper soaked with Kanamycin solutions (100–500 mg/l). Opening of cotyledonary leaves and growth of wild type seedlings reduced considerably in Kanamycin concentrations ≥200 mg/l, than the transgenic, thereby differentiating from the transgenic. The transgenic confirmation experiments were repeated in subsequent T8, T9 and T10 generations with the same results. Thus, the Kanamycin resistance based transgenic confirmation methods are of wide applications for research community, as well as for enforcing bio-safety regulations, especially during the containment and confined field trials.

Workforce planning in an era of health care reform is a challenge as both delivery systems and patient demographics change. Current workforce projections are based on a future health care system that is either an identified "ideal" or a modified version of the existing system. The desire to plan for such an "ideal system," however, may threaten access to necessary services if it does not come to fruition or is based on theoretical rather than empirical data.Historically, workforce planning that concentrated only on an "ideal system" has been centered on incorrect assumptions. Two examples of such failures presented in the 1980s when the Graduate Medical Education National Advisory Committee recommended a decrease in the physician workforce on the basis of predetermined "necessary and appropriate" services and in the 1990s, when planners expected managed care and health maintenance organizations to completely overhaul the existing health care system. Neither accounted for human behavior, demographic changes, and actual demand for health care services, leaving the nation ill-prepared to care for an aging population with chronic disease.In this article, the authors argue that workforce planning should begin with the current system and make adjustments based on empirical data that accurately reflect current trends. Actual health care use patterns will become evident as systemic changes are realized-or not-over time. No single approach will solve the looming physician shortage, but the danger of planning only for an ideal system is being unprepared for the actual needs of the population.

Jatropha curcas L. is gaining importance as a potential energy crop. However, lack of sufficient numbers of molecular markers hinder current research on crop improvement in Jatropha. The expressed sequences tags (EST) sequences deposited in public databases, offers an excellent opportunity to identify simple sequence repeats (SSRs) through data mining, for further research on molecular breeding. In the present study 42,477 ESTs of J. curcas were screened, out of which 5,673 SSRs were identified with 48.8 % simple (excluding mononucleotide repeats) and 52.2 % compound repeat motifs. Amongst these repeat motifs, dinucleotide repeats were abundant (26.5 %), followed by trinucleotide (23.1 %) and tetranucleotide repeats (0.8 %). From these microsatellites, 32 EST-SSR (genic microsatellite) primer pairs were designed. These primers were used to analyze the genetic diversity among 42 accessions collected from different parts of India. Out of the 32 EST-SSR primers, 24 primer pairs exhibited polymorphism among the genotypes, with amplicons varying from one to eight, giving an average of 2.33 alleles per polymorphic marker. Polymorphic information content value ranged from 0.02 to 0.5 with an average of 0.402 indicating moderate level of informativeness within these EST-SSRs markers. The EST-SSR markers developed here will serve as a valuable resource for genetic studies, like linkage mapping, diversity analysis, quantitative trait locus/association mapping, and molecular breeding. The current study also revealed low diversity in the screened Indian Jatropha germplasm. Therefore, the future efforts must be made to broaden the gene pool of Jatropha for the creation of genetic diversity that can be further used for crop improvement through breeding.

Pseudomonas syringae pv. phaseolicola is a major plant pathogen causing halo blight disease and has world-wide importance. The emerging post-genomics field of agrigenomics, together with the availability of whole genome sequences of a number of pathogens and host organisms, offer the promise for identification of potential drug targets using sequence comparison approaches. On the other hand, lack of gene expression data for most of the phytopathogenic microbes still remains a formidable barrier. The present study aimed at the prediction of drug targets in Pseudomonas syringae pv. phaseolicola by exploiting the knowledge of Codon Usage bias for gene expression subtractively, supported by gene expression analysis and sequence comparisons. Based on screening of the Database of Essential Genes using blastx, 158 of the total 5172 genes of P. syringae pv. phaseolicola were enlisted as vitally essential genes. Similarity search for these 158 essential genes against available host-plant sequences (Phaseolous vulgaris) led to the identification of homologues of 21 genes in the host genome, thus leaving behind a subset of 137 genes. Expression analysis of these 137 genes using RSCU(gene,) validated by microarray gene expression data suggested 22 genes had higher expression levels in the cell, and therefore their products have been identified as putative drug targets. The gene ontology analysis of these 22 genes revealed their indispensable roles in pivotal metabolic pathways of P. syringae pv. phaseolicola. Upon comparison of the sequences of these genes with other soil bacteria, we identified two genes that were unique to P. syringae pv. phaseolicola. The products of these genes can potentially be utilized for drug development so as to control the halo blight disease and thereby accelerate translation research in the nascent field of agrigenomics.

Cross species hybridization is an attractive option to understand the regulatory mechanisms and transcriptional networks underlying a biological activity, particularly for a non model organism. We have used the same approach to idenitfy cold responsive differentially expressed genes (DEGs) in Lepidium latifolium using Arabidopsis thaliana cDNA array chip. In all, 1315 DEGs were discovered. Putative functions of these cold-responsive genes were explored based on annotations using TAIR database and Blast2GO. Most of the up-regulated gene products were mapped to nucleus (20%) and plastids (15%) showing higher activities as DNA binding proteins, i.e., transcription factors. In all, 45 genes encoding transcription factors were identified belonging most frequently to AP2 family proteins (Seven genes) followed by NAM domain containing proteins (Five genes). Transcription factor binding sites (TFBS) were predicted for 1,100 genes belonging to 78 clusters of co-expressed genes. Interestingly, 50% of these regulatory sites contained a microsatellite repeat. Considering, Lepidium to be a native of high altitude cold regions, the genes identified in this study can be of wide importance to agricultural biotechnologists.

NAC genes contribute to enhance survivability of plants under conditions of environmental stress and in secondary growth of the plants, thereby building biomass. Thus, genetic transformation of plants using NAC genes provides a possibility to tailor made biofuel plants. Over-expression studies have indicated that NAC family genes can provide tolerance to various biotic and abiotic stresses, either by physiological or biochemical changes at the cellular level, or by affecting visible morphological and anatomical changes, for example by development of lateral roots in a number of plants. Over-expression of these genes also work as triggers for development of secondary cell walls. In our laboratory, we have observed a NAC gene from Lepidium latifolium contributing to both enhanced biomass as well as cold stress tolerance of model plants tobacco. Thus, we have reviewed all the developments of genetic engineering using NAC genes which could enhance the traits required for biofuel plants, either by enhancing the stress tolerance or by enhancing the biomass of the plants. KeywordsNAC, Genetically engineered plants, Abiotic stress tolerance, Secondary growth, Cell wall synthesis, Biomass

Lepidium latifoliumL. (Brassicaceae) grows successfully in a high-altitude cold arid environment. Little molecular data are available for this plant despite its immense ecological importance as a cold- and drought-adapted species. We used a novel approach to identify microsatellite regions using genome walker libraries, called as Random Scans at Microsatellite Regions (RaSMiR), and implemented them on genotypes collected from relatively different topographical conditions within a small geographical area. The success rate of finding a microsatellite sequence using this methodology was 100%, and on developing the RaSMiR technique itself as a molecular marker, 230 electrophoretic bands were obtained using 13 different RaSMiR primers in combination with a microsatellite sequence primer. On an average, 17 bands were obtained for each primer. The electrophoretic profiles generated by RaSMiR markers were distinct from those produced by inter-simple sequence repeat markers. This information has been documented as a dominant marker data, and has been used to construct a neighbour-joining tree that successfully distinguished all genotypes. RaSMiR is an attractive approach for the development of unique and informative microsatellites, or for genome scanning directly as a molecular marker that can potentially be employed for the estimation of genetic diversity or to identify polymorphic loci involved in adaptations particularly in the non-model species, for which sufficient genomic data are not available.

The high altitude regions owing to their unique climatic and geographic differentiation have specific agro-technological requirements, different from the ones employed elsewhere for vegetable production. The major high altitude abiotic stresses includes cold, frost, drought, salinity, low oxygen, high wind velocity and intense UV radiations etc. that negatively influence the survival, biomass production and yields of vegetable crops 1 up to 70 per cent. Since tolerance to these stresses is multigenic and quantitative in nature, a massive challenge exists to understand the key molecular mechanisms for advanced selective breeding purposes. A lot of efforts made to identify the suitable varieties/hybrids and cultivation practices, the complexity of trait of tolerance to abiotic stresses remains the major limitation in overcoming these barriers for better agricultural productivity of high altitude regions. Also, the mechanisms by which plants perceive environmental signals and further their transmission to cellular machinery to activate adaptive responses is of critical importance for the development of rational breeding and transgenic strategies to impart abiotic stress tolerance in vegetable crops 2 . Conventional breeding methods have met with limited success in improving the abiotic stress tolerance of important vegetables while biotechnology offers new and highly precise strategies that can be used to develop transgenics crop plants with improved tolerance to abiotic stress that would certainly enhance the productivity of vegetable crops at high altitude regions 3 . Although, recent advancements in genomics, transcriptomics and regulomics have enriched our understanding on stress physiology of plants, and it is hoped that this will contribute to the development of tolerant genotypes for such regions. Timely modulation of specific sets of genes is critical for

Hippophae salicifolia D. Don (Seabuckthorn) grows in stressful environment of high altitude under conditions of low temperature and low availability of water. We have studied gender based differences in physiochemical response to cold stress in male and female plants of Seabuckthorn. After 24 h of cold stress about 32 and 66 % higher electrolyte leakage (EL) was recorded in male and female plants respectively. Relative water content (RWC) at the end of 24 h stress was higher in male plants (~64 %) compared to female plants (~60 %). Proline content in leaf samples of cold stressed male and female plants also increased upon cold stress. After 24 h about 2.7 fold higher amount of proline was assessed in male and female in comparison to control plants. Similarly, about two fold increase in the specific activities of superoxide dismutase (SOD) and catalase was also observed upon cold stress in male and female plants. These findings have important inferences for community of molecular biologists exploring seabuckthorn genome for agronomically important genes.

Characterization of occurrence, density and motif sequence of tandem repeats in the transcribed regions is helpful in understanding the functional significance of these repeats in the modern genomes. We analyzed tandem repeats present in expressed sequences of thirteen species belonging to genera Capsicum, Nicotiana, Petunia and Solanum of family Solanaceae and the genus Coffea of Rubiaceae to investigate the propagation and evolutionary sustenance of these repeats. Tandem repeat containing sequences constituted 1.58% to 7.46% of sequences analyzed. Tandem repetitions of size 2, 15, 18 and 21 bp motifs were more frequent. Repeats with unit sizes 21 and 22 bp were also abundant in genomic sequences of potato and tomato. While mutations occurring in these repeats may alter the repeat number, genomes adjust to these changes by keeping the translated products unaffected. Surprisingly, in majority of the species under study, tandem repeat motif length did not exceed 228 bp. Conserved tandem repeat motifs of sizes 180, 192 and 204 bp were also abundant in the genomic sequences. Our observations lead us to propose that these tandem repeats are actually remnants of ancestral megasatellite repeats, which have split into multiple repeats due to frequent insertions over the course of evolution.

Phylogenetic footprinting is a method for the discovery of regulatory elements in a set of homologous regulatory regions, usually collected from multiple species. It does so by identifying the best conserved motifs in those homologous regions. There are two popular sets of methods-alignment-based and motif-based, which are generally employed for phylogenetic methods. However, serious efforts have lacked to develop a tool exclusively for phylogenetic footprinting, based on either of these methods. Nevertheless, a number of software and tools exist that can be applied for prediction of phylogenetic footprinting with variable degree of success. The output from these tools may get affected by a number of factors associated with current state of knowledge, techniques and other resources available. We here present a critical apprehension of various phylogenetic approaches with reference to prokaryotes outlining the available resources and also discussing various factors affecting footprinting in order to make a clear idea about the proper use of this approach on prokaryotes.

Seed dormancy is an important limiting factor in exploitation of an economically important species to its fullest. Hippophae salicifolia D. Don (seabuckthorn), a rich source of medicinal metabolites shows both exogenous and endogenous dormancy. Evidently, we recorded a high seed viability (94 %) but poor germination (22 %) of untreated seeds. We applied different pre-sowing seed priming treatments including NaCl (50, 100, 200, 500 mM), KNO3 (0.1, 0.2, 0.3 %), Thiourea (1, 2, 3 %), GA3 (100, 250, 500 mg/L), Sulphuric acid (98 %) and cold (4 °C) and warm water (65 °C) stratifications to explore improvements in its germination percentage, if any. We found KNO3 (0.1 %) and Thiourea (1 %) treatments to be superior to other methods for enhancement of mean seed germination percentage of H. salicifolia. Considering the practical applicability and cost effectiveness, these treatments can be applied to overcome seed dormancy and recommended for mass multiplication through seeds of H. salicifolia.

Background— Even as the burden of cardiovascular disease in the United States is increasing as the population grows and ages, the number of active cardiothoracic surgeons has fallen for the first time in 20 years. Meanwhile, the treatment of patients with coronary artery disease continues to evolve amid uncertain changes in technology. This study evaluates current and future requirements for cardiothoracic surgeons in light of decreasing rates of coronary artery bypass grafting procedures. Methods and Results— Projections of supply and demand for cardiothoracic surgeons are based on analysis of population, physician office, hospital, and physician data sets to estimate current patterns of healthcare use and delivery. Using a simulation model, we project the future supply of cardiothoracic surgeons under alternative assumptions about the number of new fellows trained each year. Future demand is modeled, taking into account patient demographics, under current and alternative use rates that include the elimination of open revascularization. By 2025, the demand for cardiothoracic surgeons could increase by 46% on the basis of population growth and aging if current healthcare use and service delivery patterns continue. Even with complete elimination of coronary artery bypass grafting, there is a projected shortfall of cardiothoracic surgeons because the active supply is projected to decrease 21% over the same time period as a result of retirement and declining entrants. Conclusion— The United States is facing a shortage of cardiothoracic surgeons within the next 10 years, which could diminish quality of care if non–board-certified physicians expand their role in cardiothoracic surgery or if patients must delay appropriate care because of a shortage of well-trained surgeons.

In 2006, after many months of consideration, the Association of American Medical Colleges (AAMC) recommended that medical schools in the United States increase their enrollment by 30% by 2015 and that residency positions be increased to accommodate the growth in U.S. medical school graduates1. This recommendation is based on the belief that there will be a substantial physician workforce shortage in the future as the economy continues to expand, physicians retire, and patients continue to demand more specialized care. How this prediction will affect the workforce dynamic of orthopaedic surgery and other specialties must be examined carefully. Orthopaedic surgery, like most specialties, has an interest in better understanding how many physicians will be required in the specialty in the future. This is not to suggest that there is one single correct number of physicians in a specialty; in fact, the medical system has proven to be highly adaptive. However, it is in the public's interest to have a distribution of orthopaedic surgeons that promotes high-quality care. Furthermore, it is in the specialty's interest that the number of physicians be sufficient to provide the services that the specialty is best qualified to perform but not so many that physicians are underutilized. Having too few orthopaedic surgeons can lead to access problems for patients and/or to less qualified providers caring for patients with particular problems. Having too many can increase competition, flatten incomes, and reduce procedural volume for individual surgeons and thereby affect quality and outcomes— not to mention possibly increasing unnecessary operations to maintain income levels. Therefore, it is beneficial to both the public and the orthopaedic surgery specialty to have a supply that is close to expected utilization. Achieving balance is easier said than done given the many factors that could influence future supply and demand. One must …

Nucleotide composition, codon usage and amino acid content are important molecular signatures that vary in different groups of organisms. AT-rich (or GC poor) hyperthermophiles have relatively been unexplored in these aspects. In this study, we have examined the compositional characteristics of AT rich genomes viz. Methanococcus jannaschii, Sulfolobus solfataricus, Sulfolobus tokodaii and Nanoarcheum equitans by their comparison with four mesophiles having similar genomic GC content. The analysis revealed a significant increase in purine content of ORFs due to increase in guanine content. Moreover, the influence of dinucleotide composition on protein thermostability was found even larger. Accordingly, increased usage of codons that are constituted of dinucleotides RR was observed. Arginine, proline, valine and tyrosine were most abundant amino acids in hyperthermophilic proteomes, and similar bias was seen when dipeptidic composition of proteins was compared. Further amino acid composition analysis of alpha helices indicates an increased usage of E, K, R and decreased usage of N and Q. Summing up, the study suggested that elevated growth temperature im- pose selective constraints at all the three molecular levels- nucleotide composition, codon usage and amino acid content.

During recent decades, microsatellites have become the most popular source of genetic markers. More recently, the availability of enormous sequence data for a large number of eukaryotic genomes has accelerated research aimed at understanding the origin and functions of microsatellites and searching for new applications. This review presents recent developments of in silico mining of microsatellites to reveal various facets of the distribution and dynamics of microsatellites in eukaryotic genomes. Two aspects of microsatellite search strategies – using a suitable search tool and accessing a relevant microsatellite database – have been explored. Judicious microsatellite mining not only helps in addressing biological questions but also facilitates better exploitation of microsatellites for diverse applications.

Microsatellites are useful tools to study the extent of divergence between two taxonomic groups that show high sequence similarity. We have compared microsatellite distribution to illustrate genetic variation between the two rice genomes, Oryza sativa L. ssp. indica and Oryza sativa L. ssp. japonica. Microsatellite distribution proved to be non random as certain regions of very high microsatellite density have been identified. Microsatellite density in the subspecies japonica was computed marginally higher than in the subspecies indica in the genomic regions compared between the two subspecies. Unexpectedly high microsatellite densities were observed in 5'-untranslated regions of genes. These regions also displayed a clear motif bias. Some of the longest microsatellite repeats were found in intron sequences. Frequency, as well as motif bias was also noted with respect to the association of microsatellites with transposable elements. Microsatellite mutability values were exemplarily estimated for 90 loci by aligning the microsatellite containing regions between the two genomes. Poor rates of finding an orthologue corresponded with high microsatellite mutability in rice. These insights are likely to play a significant role in selecting microsatellite loci to be used in molecular breeding and studying evolutionary dynamics of the two subspecies.

Background: Workforce studies may be disconnected from the policies that affect supply and demand for health professionals. Sporadic review of the physician workforce in the last century has led to wide swings in perception about its adequacy. However, workforce research has influenced federal policy as well as the policies of institutions responsible for training and regulation of physicians. This discussion is intended to address workforce issues in the context of public policy at the federal level. It is also intended to serve as a starting point for new approaches to shaping workforce policy. Discussion: The supply of and demand for physicians and other health professionals are affected by a number of factors that may or may not be under the control of policymakers and health professionals themselves. Productivity, practice patterns, the aging of the workforce and patients, and other major determinants are only minimally affected by most government policy. Despite several attempts throughout the 1980s and 1990s, demand for health care has been particularly difficult to control for policymakers. In contrast to the United States, most developed nations are extensively involved in the planning of the healthcare workforce, including the specialty mix, the number of physicians, and the number of other health professionals. Conclusion: There are many barriers to successful workforce policy. Successful public policy change often involves multiple stakeholders, in and out of government. The task before those concerned about workforce issues is to educate policymakers about how changes in the physician workforce will affect cost, access, and quality, and to impress upon them that serious efforts to improve quality of care and reduce costs will not be effective unless qualified physicians are there to provide that care.

Completion of rice genome sequencing has necessitated identification of transcripts encoded by the genome and their corresponding functions. In the present study we have catalogued cytoplasmic ribosomal protein complement of rice genome corresponding to 68 families. Mining TIGR (The Institute for Genomic Research) rice database and unigene sequences available from National Centre for Biotechnology Information (NCBI) produced a non-redundant set of these sequences. This resulted in identification of 209 candidate r-proteins of which 22 have not been reported previously. The number of genes per family ranges from 1 to 8 distributed throughout the genome with maximum occurrence at chromosome 7. Mapping of r-proteins on BAC clones revealed several small clusters of genes. Unigene sequences corresponding to most of the reported r-proteins were identified indicating these genes are being expressed.

Analyses at the county level show lower mortality rates where there are more primary care physicians, but this is not the case for specialist supply. These findings confirm those of previ...