Classification of substances by health hazard using deep neural networks and molecular electron densities.

In this paper we present a method that allows leveraging 3D electron density information to train a deep neural network pipeline to segment regions of high, medium and low electronegativity and classify substances as health hazardous or non-hazardous. We show that this can be used for use-cases such as cosmetics and food products. For this purpose, we first generate 3D electron density cubes using semiempirical molecular calculations for a custom European Chemicals Agency (ECHA) subset consisting of substances labelled as hazardous and non-hazardous for cosmetic usage. Together with their 3-class electronegativity maps we train a modified 3D-UNet with electron density cubes to segment reactive sites in molecules and classify substances with an accuracy of 78.1%. We perform the same process on a custom food dataset (CompFood) consisting of hazardous and non-hazardous substances compiled from European Food Safety Authority (EFSA) OpenFoodTox, Food and Drug Administration (FDA) Generally Recognized as Safe (GRAS) and FooDB datasets to achieve a classification accuracy of 64.1%. Our results show that 3D electron densities and particularly masked electron densities, calculated by taking a product of original electron densities and regions of high and low electronegativity can be used to classify molecules for different use-cases and thus serve not only to guide safe-by-design product development but also aid in regulatory decisions. SCIENTIFIC CONTRIBUTION: We aim to contribute to the diverse 3D molecular representations used for training machine learning algorithms by showing that a deep learning network can be trained on 3D electron density representation of molecules. This approach has previously not been used to train machine learning models and it allows utilization of the true spatial domain of the molecule for prediction of properties such as their suitability for usage in cosmetics and food products and in future, to other molecular properties. The data and code used for training is accessible at https://github.com/s-singh-ivv/eDen-Substances .

Pneumothorax Secondary to a Traumatic Brazilian Jiu-Jitsu Injury.

A pneumothorax is a medical condition characterized by the presence of free air in the pleural cavity. Pneumothorax can be classified as spontaneous, traumatic, or iatrogenic. Spontaneous pneumothorax sustained from a jiu-jitsu-induced blunt trauma has not been described in any sports literature. This case report involves a 26-year-old male athlete who presented to the emergency room complaining of right-sided chest pain in the recumbent position and shortness of breath upon exertion. Breath sounds were diminished on the right with hyper resonance to percussion. Inspection of the chest revealed diffuse erythema on the right side. A chest X-ray revealed a right tension pneumothorax that was treated with a 20-French chest tube. This report aims to highlight the importance of recognizing the possibility of pneumothorax in jiu-jitsu athletes, implementing early treatment, and exploring potential causes of pneumothorax in otherwise healthy individuals.

Quick insights into whisky - investigating rapid and efficient methods for sensory evaluation and chemical analysis.

The evaluation of aroma properties of beverages, both analytically as well as with human sensory studies, is a challenging task and most often related to time- and cost-intensive analyses. Whisky is a spirit offering a wide variety of aroma impressions caused by a complex mixture of aroma active compounds. In the present study, methods for the efficient evaluation of aroma characteristics are evaluated for 16 whisky samples of different origins (Scotch and American). Rate all that apply (RATA) was applied as a rapid method for the sensory evaluation of whiskies. Sensory evaluation of the samples led to the determination of eight significant aroma attributes: caramel-/cream caramel-/toffee-like, vanilla-like, (canned) peach-like, phenolic, smoky, fruity, flowery and (fermented) apple-/cider-like. Chemical analysis was conducted by stir bar sorptive extraction (SBSE) coupled to gas chromatography-mass spectrometry in combination with an in-house data processing tool for semi-automated analyte detection. Through chemical analysis of the whisky samples and automated compound detection, we report over 200 mostly aroma-active volatiles. To test both approaches for their potential for sample classification, a simple classification problem (Scotch vs. American) was applied. Linear discriminant analysis (LDA) indicates both that sensory evaluation by RATA (97.86%) and the applied analytical procedure (96.94%) are suitable for the distinction between the two whisky types. In both approaches, potential markers were determined for the classification. These investigations build a solid foundation for the implementation of a versatile platform facilitating rapid and efficient aroma evaluation in various foodstuffs and beverages.

Nanoparticle-Shielded dsRNA Delivery for Enhancing RNAi Efficiency in Cotton Spotted Bollworm Earias vittella (Lepidoptera: Nolidae).

The spotted bollworm Earias vittella (Lepidoptera: Nolidae) is a polyphagous pest with enormous economic significance, primarily affecting cotton and okra. However, the lack of gene sequence information on this pest has a significant constraint on molecular investigations and the formulation of superior pest management strategies. An RNA-seq-based transcriptome study was conducted to alleviate such limitations, and de novo assembly was performed to obtain transcript sequences of this pest. Reference gene identification across E. vittella developmental stages and RNAi treatments were conducted using its sequence information, which resulted in identifying transcription elongation factor (TEF), V-type proton ATPase (V-ATPase), and Glyceraldehyde -3-phosphate dehydrogenase (GAPDH) as the most suitable reference genes for normalization in RT-qPCR-based gene expression studies. The present study also identified important developmental, RNAi pathway, and RNAi target genes and performed life-stage developmental expression analysis using RT-qPCR to select the optimal targets for RNAi. We found that naked dsRNA degradation in the E. vittella hemolymph is the primary reason for poor RNAi. A total of six genes including Juvenile hormone methyl transferase (JHAMT), Chitin synthase (CHS), Aminopeptidase (AMN), Cadherin (CAD), Alpha-amylase (AMY), and V-type proton ATPase (V-ATPase) were selected and knocked down significantly with three different nanoparticles encapsulated dsRNA conjugates, i.e., Chitosan-dsRNA, carbon quantum dots-dsRNA (CQD-dsRNA), and Lipofectamine-dsRNA conjugate. These results demonstrate that feeding nanoparticle-shielded dsRNA silences target genes and suggests that nanoparticle-based RNAi can efficiently manage this pest.

OWSum: algorithmic odor prediction and insight into structure-odor relationships.

We derived and implemented a linear classification algorithm for the prediction of a molecule's odor, called Olfactory Weighted Sum (OWSum). Our approach relies solely on structural patterns of the molecules as features for algorithmic treatment and uses conditional probabilities combined with tf-idf values. In addition to the prediction of molecular odor, OWSum provides insights into properties of the dataset and allows to understand how algorithmic classifications are reached by quantitatively assigning structural patterns to odors. This provides chemists with an intuitive understanding of underlying interactions. To deal with ambiguities of the natural language used to describe odor, we introduced descriptor overlap as a metric for the quantification of semantic overlap between descriptors. Thus, grouping of descriptors and derivation of higher-level descriptors becomes possible. Our approach poses a large leap forward in our capabilities to understand and predict molecular features.

Cultured meat - a patentometric analysis.

Cultured meat (CM) has emerged as a breakthrough technology, which has the potential to minimise future negative impacts associated with the rapidly growing world population; such as serious environmental, sustainability, global public health, and animal welfare concerns. Recently numerous researchers in academia and companies have been putting significant efforts into scientific and translational development in this field. Since various pillars of CM manufacturing hold substantial translational potential, there has been a steady interest in filing patent applications globally in the past decade. For the first time, we have performed a patentometric analysis for cultured meat by highlighting the current patents landscape, determining prolific organisations and geographical locations active in the area, and identifying the overall outlook for the field. In total, > 190 patents published in the English language were collected from 1997 to 2020 for the analysis. Relevant data were extracted from the patents, including technology focus, country of origin, assignees, priority date, cooperative patent classification (CPC) code, to analyse the current patent landscape.

Doublesex homolog is sex-specifically spliced and governs the sexual differentiation process in the whitefly Bemisia tabaci AsiaII-1.

The silverleaf whitefly Bemisia tabaci is one of the most destructive of crop pests globally. In Northern India cotton is predominately infested by the Asia II-1 species of B. tabaci. Though B. tabaci exhibits unique haplodiploidy in its reproductive behavior, to date very little is known regarding its sex determination mechanism. Here, an in-depth characterization of the AsiaII-1 doublesex (Btdsx) gene, which has been implicated in sex determination in B. tabaci, indicates the inclusion of six exons and five introns. The pre-mRNA is shown to sex-specifically splice, producing four male isoforms and one female isoform. These BtDsx proteins share common DNA binding (OD1) domains whereas they differ at their C-termini. RT-qPCR analysis revealed a significantly higher expression of Btdsx in female adults compared to that in male adults and earlier developmental stages. Functional characterization of Btdsx through RNA interference (RNAi) resulted in a significant reduction in its expression in both sexes. Btdsx knockdown concomitantly resulted in up-regulation of the expression of vitellogenin (vg) and vitellogenin receptor (vgr) genes in males and their down-regulation in females. Btdsx knockdown followed by mating resulted in reduced fecundity and percent egg hatching; however, no impact was observed on the female: male ratios in the progeny obtained from knockdown parents.

Novel application of intravascular lithotripsy in stent under-expansion: A single-center experience.

INTRODUCTION: Stent under-expansion due to calcification is associated with a less durable result. The development of intravascular lithotripsy (IVL) has provided clinicians with a readily available, simple-to-use treatment option for coronary calcification, but the use of IVL within a previously stented segment is currently off-license. There are, however, developing data suggesting that the use of IVL can be an effective treatment option for patients with calcific stent under-expansion. METHOD: This was a single-center study of all patients treated with IVL for calcific stent under-expansion between January 2019 and June 2021. The impact of IVL on quantitative coronary angiography (QCA) stenosis and on the minimal stent area (MSA) derived from intracoronary imaging were recorded. The presence of periprocedural complications and adverse cardiovascular events was obtained from the clinical record during the study timeframe. RESULTS: Thirty-nine patients underwent IVL for calcific stent under-expansion during the study time frame with one patient treated with more than one lesion in the same session. In all lesions, there was an improvement in the QCA stenosis with 37 (92.5%) having a residual stenosis of ≤30%. The mean QCA stenosis pre-IVL was 68 ± 21% and following IVL the mean QCA was 18 ± 9% (p < 0.001). In all lesions, there was an improvement in the MSA, with 26 (92.9%) achieving an MSA of more than 4.5 mm2 . The mean MSA pre-IVL was 3.88 ± 1.51 mm2 and following IVL the mean MSA was 7.41 ± 2.34 mm2 (p < 0.001). There were no major procedural complications. Over a mean follow-up of 506 ± 277 days, one patient died from ventricular arrhythmia but there were no other major adverse cardiovascular events. CONCLUSION: This single-center study demonstrates that IVL is a safe and effective treatment for calcific stent under-expansion with good medium-term results.

Unified trade-off optimization of quantum harmonic Otto engine and refrigerator.

We investigate quantum Otto engine and refrigeration cycles of a time-dependent harmonic oscillator operating under the conditions of maximum Ω function, a trade-off objective function which represents a compromise between energy benefits and losses for a specific job, for both adiabatic and nonadiabatic (sudden) frequency modulations. We derive analytical expressions for the efficiency and coefficient of performance of the Otto cycle. For the case of adiabatic driving, we point out that in the low-temperature regime, the harmonic Otto engine (refrigerator) can be mapped to Feynman's ratchet and pawl model which is a steady-state classical heat engine. For the sudden switch of frequencies, we obtain loop-like behavior of the efficiency-work curve, which is characteristic of irreversible heat engines. Finally, we discuss the behavior of cooling power at maximum Ω function.

CRISPR/Cas tool designs for multiplex genome editing and its applications in developing biotic and abiotic stress-resistant crop plants.

Crop plants are prone to several yield-reducing biotic and abiotic stresses. The crop yield reductions due to these stresses need addressing to maintain an adequate balance between the increasing world population and food production to avoid food scarcities in the future. It is impossible to increase the area under food crops proportionately to meet the rising food demand. In such an adverse scenario overcoming the biotic and abiotic stresses through biotechnological interventions may serve as a boon to help meet the globe's food requirements. Under the current genomic era, the wide availability of genomic resources and genome editing technologies such as Transcription Activator-Like Effector Nucleases (TALENs), Zinc Finger Nucleases (ZFNs), and Clustered-Regularly Interspaced Palindromic Repeats/CRISPR-associated proteins (CRISPR/Cas) has widened the scope of overcoming these stresses for several food crops. These techniques have made gene editing more manageable and accessible with changes at the embryo level by adding or deleting DNA sequences of the target gene(s) from the genome. The CRISPR construct consists of a single guide RNA having complementarity with the nucleotide fragments of the target gene sequence, accompanied by a protospacer adjacent motif. The target sequence in the organism's genome is then cleaved by the Cas9 endonuclease for obtaining a desired trait of interest. The current review describes the components, mechanisms, and types of CRISPR/Cas techniques and how this technology has helped to functionally characterize genes associated with various biotic and abiotic stresses in a target organism. This review also summarizes the application of CRISPR/Cas technology targeting these stresses in crops through knocking down/out of associated genes.

Introgressing cry1Ac for Pod Borer Resistance in Chickpea Through Marker-Assisted Backcross Breeding.

The gram pod borer Helicoverpa armigera is a major constraint to chickpea (Cicer arietinum L.) production worldwide, reducing crop yield by up to 90%. The constraint is difficult to overcome as chickpea germplasm including wild species either lacks pod borer resistance or if possessing resistance is cross-incompatible. This study describes conversion of elite but pod borer-susceptible commercial chickpea cultivars into resistant cultivars through introgression of cry1Ac using marker-assisted backcross breeding. The chickpea cultivars (PBG7 and L552) were crossed with pod borer-resistant transgenic lines (BS 100B and BS 100E) carrying cry1Ac that led to the development of BC1F1, BC1F2, BC1F3, BC2F1, BC2F2, and BC2F3 populations from three cross combinations. The foreground selection revealed that 35.38% BC1F1 and 8.4% BC1F2 plants obtained from Cross A (PBG7 × BS 100B), 50% BC1F1 and 76.5% BC1F2 plants from Cross B (L552 × BS 100E), and 12.05% BC2F2 and 82.81% (average) BC2F3 plants derived from Cross C (PBG7 × BS 100E) carried the cry1Ac gene. The bioassay of backcross populations for toxicity to H. armigera displayed up to 100% larval mortality. BC1F1 and BC1F2 populations derived from Cross B and BC2F3 population from Cross C segregated in the Mendelian ratio for cry1Ac confirmed inheritance of a single copy of transgene, whereas BC1F1 and BC1F2 populations obtained from Cross A and BC2F2 population from Cross C exhibited distorted segregation ratios. BC1F1 plants of Cross A and Cross B accumulated Cry1Ac protein ranging from 11.03 to 11.71 µgg-1 in leaf tissue. Cry1Ac-positive BC2F2 plants from Cross C demonstrated high recurrent parent genome recovery (91.3%) through background selection using SSR markers and phenome recovery of 90.94%, amongst these 30% plants, were homozygous for transgene. The performance of BC2F3 progenies derived from homozygous plants was similar to that of the recurrent parent for main agronomic traits, such as number of pods and seed yield per plant. These progenies are a valuable source for H. armigera resistance in chickpea breeding programs.

Pervasive Endosymbiont Arsenophonus Plays a Key Role in the Transmission of Cotton Leaf Curl Virus Vectored by Asia II-1 Genetic Group of Bemisia tabaci.

Insects often coevolved with their mutualistic partners such as gut endosymbionts, which play a key in the physiology of host. Studies on such interactions between Bemisia tabaci and its primary and secondary endosymbionts have gained importance due to their indispensable roles in the biology of this insect. Present study reports the predominance of two secondary endosymbionts, Arsenophonus and Cardinium in the Asia II-1 genetic group of whitefly and elucidates their role in the transmission of its vectored Cotton leaf curl virus. Selective elimination of endosymbionts was optimized using serial concentration of ampicillin, chloramphenicol, kanamycin, tetracycline, and rifampicin administered to viruliferous whiteflies through sucrose diet. Primary endosymbiont, Portiera was unresponsive to all the antibiotics, however, rifampicin and tetracycline at 90 µg/ml selectively eliminated Arsenophonus from the whitefly. Elimination of Arsenophonus resulted in significant decrease in virus titer from viruliferous whitefly, further the CLCuV transmission efficiency of these whiteflies was significantly reduced compared to the control flies. Secondary endosymbiont, Cardinium could not be eliminated completely even with higher concentrations of antibiotics. Based on the findings, Arsenophonus plays a key role in the retention and transmission of CLCuV in the Asia II-1 genetic group of B. tabaci, while the role of Cardinium could not be established due to its unresponsiveness to antibiotics.

Design and Evaluation of MeVisLab Networks for Co-Registration and Cropping of Positron Emission Tomography/Computed Tomography Scans.

Objective: The aim of the present study was to design and evaluate two MeVisLab networks, one for co-registration of positron emission tomography/computed tomography (PET/CT) images and second for cropping the co-registered PET/CT images. Materials and Methods: Two MeVisLab networks, one to co-register and export PET/CT DICOM images and second for cropping the co-registered PET/CT images were designed using different modules of registration toolkit MERIT. One hundred and twenty-five PET/CT studies were exported from Siemens and GE scanners in DICOM format. These images were co-registered and cropped with our designed networks. The images co-registered with our network were compared visually with the co-registered images of same PET/CT studies on vendor provided workstations by an experienced nuclear medicine physician (NMP). The perfection of the cropping of co-registered images was also assessed visually. Results: Visually, NMP found all 125 images co-registered using the network designed in our study similar to the co-registered images of vendor provided workstations. Furthermore, the cropping of all co-registered images was perfectly done by our network. Conclusion: Two MeVisLab networks designed and evaluated in the present study can be used for co-registration of PET/CT DICOM images and cropping the co-registered PET/CT images.

Identification and validation of stage-specific reference genes for gene expression analysis in Callosobruchus maculatus (Coleoptera: Bruchidae).

In light of a number of recent studies highlighting the increasing research interest in bruchids, it is crucial to validate suitable reference genes that could be used in quantitative gene expression studies. Callosobruchus maculatus is a serious pest of stored grains and field legumes in which reference genes have not been assessed and validated to date. The present study aimed to identify and validate reference genes in different developmental stages of C. maculatus shortlisted from commonly used reference genes such as VATPase, TRIP12, TBP, TF11D, ACTIN, GST, ANNEXIN, PTCD3, RPL32, and ß -Tub in various insects. Dedicated algorithms like GeNorm, NormFinder, and BestKeeper were used to analyze the stability of these candidate genes, which revealed GST for third instar, ANNEXIN and PTCD3 for the fourth instar, TF11D and VATPase for male pupa, RPL32 and ß-tub for female pupa, ß-tub and TBP for adult male and VATPase and GST for adult females as suitable reference genes for expression studies in C. maculatus. The final comprehensive ranking using RefFinder identified GST and TBP as the best reference genes for all the developmental stages of C. maculatus. To the best of our knowledge, this is the first report which evaluates and validates stable reference genes in C. maculatus. The information of stage-specific gene expression, generated in this study will be useful for future molecular, physiological, and biochemical studies on C. maculatus and other closely related bruchids.

Scaffolds for the manufacture of cultured meat.

The cultured meat market has been growing at an accelerated space since the first creation of cultured meat burger back in 2013. Substantial efforts have been made to reduce costs by eliminating serum in growth media and improving process efficiency by employing bioreactors. In parallel, efforts are also being made on scaffolding innovations to offer better cells proliferation, differentiation and tissue development. So far, scaffolds used in cultured meat research are predominantly collagen and gelatin, which are animal-derived. To align with cell-based meat vision i.e. environment conservation and animal welfare, plant-derived biomaterials for scaffolding are being intensively explored. This paper reviews and discusses the advantages and disadvantages of scaffold materials and potential scaffolding related to scale-up solution for the production of cultured meat.

The unspoken benefit of participation in a clinical trial.

BACKGROUND: Publicly funded trials do not usually offer financial incentives to volunteers. An intensive level of medical care could act as an additional motivator for participation. Our aim was to establish whether patients may draw any clinical benefit from volunteering in a clinical trial. METHODS: We analysed the recruitment process of a phase II randomised controlled trial, the Inorganic Nitrate in Angina Study. RESULTS: Two-hundred and thirteen patients with a history of stable angina and who had been under at least annual primary care review were screened for participation by history taking, examination, 12-lead electrocardiography, treadmill test and echocardiography. Thirty-five (16.4%) patients were found to have significant unstable or new clinical pathology, requiring urgent clinical attention. We identified 17 (7.9%) patients with unstable angina. Furthermore, we found new undiagnosed pathologies: amyloidosis in two (0.9%), hypertrophic cardiomyopathy in two (0.9%), left ventricular systolic dysfunction (ejection fraction <45%) in three (1.4%), left ventricular thrombus in one (0.4%), significant valvular disease in five (2.4%) and arrhythmias in six (2.8%). CONCLUSION: Compared with routine care, patients screened for a clinical trial may come under an increased level of scrutiny that may affect their clinical management. This may act as additional motivator to attract patients to future studies.

Next Generation dsRNA-Based Insect Control: Success So Far and Challenges.

RNA interference (RNAi) is a method of gene silencing where dsRNA is digested into small interfering RNA (siRNA) in the presence of enzymes. These siRNAs then target homologous mRNA sequences aided by the RNA-induced silencing complex (RISC). The mechanism of dsRNA uptake has been well studied and established across many living organisms including insects. In insects, RNAi is a novel and potential tool to develop future pest management means targeting various classes of insects including dipterans, coleopterans, hemipterans, lepidopterans, hymenopterans and isopterans. However, the extent of RNAi in individual class varies due to underlying mechanisms. The present review focuses on three major insect classes viz hemipterans, lepidopterans and coleopterans and the rationale behind this lies in the fact that studies pertaining to RNAi has been extensively performed in these groups. Additionally, these classes harbour major agriculturally important pest species which require due attention. Interestingly, all the three classes exhibit varying levels of RNAi efficiencies with the coleopterans exhibiting maximum response, while hemipterans are relatively inefficient. Lepidopterans on the other hand, show minimum response to RNAi. This has been attributed to many facts and few important being endosomal escape, high activity dsRNA-specific nucleases, and highly alkaline gut environment which renders the dsRNA unstable. Various methods have been established to ensure safe delivery of dsRNA into the biological system of the insect. The most common method for dsRNA administration is supplementing the diet of insects via spraying onto leaves and other commonly eaten parts of the plant. This method is environment-friendly and superior to the hazardous effects of pesticides. Another method involves submergence of root systems in dsRNA solutions and subsequent uptake by the phloem. Additionally, more recent techniques are nanoparticle- and Agrobacterium-mediated delivery systems. However, due to the novelty of these biotechnological methods and recalcitrant nature of certain crops, further optimization is required. This review emphasizes on RNAi developments in agriculturally important insect species and the major hurdles for efficient RNAi in these groups. The review also discusses in detail the development of new techniques to enhance RNAi efficiency using liposomes and nanoparticles, transplastomics, microbial-mediated delivery and chemical methods.

Differential expression of gut protein genes and population density of Arsenophonus contributes to sex-biased transmission of Bemisia tabaci vectored Cotton leaf curl virus.

Whitefly, Bemisia tabaci (Gennadius) is an important pest of cotton causing direct damage as sap feeder and vector of Cotton leaf curl virus (CLCuV). Previous few studies suggest that female whiteflies are more efficient vector of begomovirusthan males, however the sex-biased transmission efficiency is still not clearly understood. Present studies with B. tabaci AsiaII-1 haplotype showed higher virus transmission efficiency of females compared to males. This variable begomovirus transmission efficiency has been related to previously identifiedkey factors associated with B. tabaci. The higher density of endosymbiont Arsenophonus and variable expression of some midgut proteins genes i.e. Cyclophilin, Knottin, Hsp40, Hsp70 may be possibly imparting higher vector competency to the females compared to males. The present studies suggest low abundance of Arsenophonus spp. as well as lower expressionof Cyclophilin genein males as compared to females. This is further supplemented by overexpression of Knottin, Hsp40, and Hsp70 genes in males compared to females and thus collectively all these factors might be playing a key role in low virus transmission efficiency of males. The relative density of Arsenophonus spp. and expression of midgut proteins genes in male and female whitefly first time enriches our understanding about sex-biased transmission efficiency of begomovirus.

Nanotechnology Facilitated Cultured Neuronal Network and Its Applications.

The development of a biomimetic neuronal network from neural cells is a big challenge for researchers. Recent advances in nanotechnology, on the other hand, have enabled unprecedented tools and techniques for guiding and directing neural stem cell proliferation and differentiation in vitro to construct an in vivo-like neuronal network. Nanotechnology allows control over neural stem cells by means of scaffolds that guide neurons to reform synaptic networks in suitable directions in 3D architecture, surface modification/nanopatterning to decide cell fate and stimulate/record signals from neurons to find out the relationships between neuronal circuit connectivity and their pathophysiological functions. Overall, nanotechnology-mediated methods facilitate precise physiochemical controls essential to develop tools appropriate for applications in neuroscience. This review emphasizes the newest applications of nanotechnology for examining central nervous system (CNS) roles and, therefore, provides an insight into how these technologies can be tested in vitro before being used in preclinical and clinical research and their potential role in regenerative medicine and tissue engineering.

Transcriptome analysis unravels RNAi pathways genes and putative expansion of CYP450 gene family in cotton leafhopper Amrasca biguttula (Ishida).

Cotton Leafhopper, Amrasca biguttula is an important pest of cotton and okra in the Indian subcontinent. Presently limited genomic/transcriptomic information is available for this insect in any of open source databases. The present study reports the first assembled and annotated de novo transcriptome of cotton leafhopper. Out of 75,551 transcripts, 39,613 CDS (Coding Sequence) were predicted with 35,282 showing positive blast hits with NCBI nr database. The Gene ontology (GO) analysis annotated 7431 CDS  with KEGG pathway categorizing these CDS into 22 different functional groups. The majority of CDS were annotated in signal transduction and transport catabolism pathways. The sequence data was screened for RNAi pathway genes and presence of 37 transcripts associated with this process confirmed the existence of robust RNAi machinery. The role of core RNAi machinery genes (Dicer-2, Ago-2, Piwi and Staufen) has been validated through dsRNA feeding studies. The data resource has also been used to identify potential RNAi targets and genes associated with insecticide detoxification specifically CYP 450 family. The current study provides a useful sequence resource which can be used to initiate molecular studies in this insect with emphasis on insecticide resistance, RNAi and functional genomics.