Effect of human β-Globin second intron on transient gene expression in mammalian cell lines

Exogenous protein expression in mammalian cells is necessary to produce therapeutic proteins and modern medical applications like developing DNA vaccines and gene therapy. This study examines the human-Globin (hBG) second intron's capacity for intron-mediated enhancement (IME) in various mammalian cell lines. Our study's main aim is to investigate the effect of the incorporation and arrangement of the second intron of the human Beta-globin gene into the pVAX-1 expression cassette on improving the expression of foreign genes. Two plasmids were constructed, one with the hBG second intron positioned upstream and the other downstream in the expression cassette. EGFP expression was evaluated at the mRNA and protein levels after transfection using Lipofectamine 2000 using One-way ANOVA analysis. Results showed that the pVAX-1 harbouring the hBG second intron did not lead to enhanced transient EGFP expression and did not exhibit Intron Mediated Enhancement (IME) in tested mammalian cell lines. Further investigations are necessary to understand factors contributing to the lack of enhancement and explore alternative intron options for optimizing foreign gene expression in cell lines

Identification and functional validation of a unique set of drought induced genes preferentially expressed in response to gradual water stress in peanut

Peanut, found to be relatively drought tolerant crop, has been the choice of study to characterize the genes expressed under gradual water deficit stress. Nearly 700 genes were identified to be enriched in subtractive cDNA library from gradual process of drought stress adaptation. Further, expression of the drought inducible genes related to various signaling components and gene sets involved in protecting cellular function has been described based on dot blot experiments. Fifty genes (25 regulators and 25 functional related genes) selected based on dot blot experiments were tested for their stress responsiveness using northern blot analysis and confirmed their nature of differential regulation under different field capacity of drought stress treatments. ESTs generated from this subtracted cDNA library offered a rich source of stress-related genes including signaling components. Additional 50% uncharacterized sequences are noteworthy. Insights gained from this study would provide the foundation for further studies to understand the question of how peanut plants are able to adapt to naturally occurring harsh drought conditions. At present functional validation cannot be deemed in peanut, hence as a proof of concept seven orthologues of drought induced genes of peanut have been silenced in heterologous N. benthamiana system, using virus induced gene silencing method. These results point out the functional importance for HSP70 gene and key regulators such as Jumonji in drought stress response

Encryption and Decryption of Images with Pixel Data Modification Using Hand Gesture Passcodes

To ensure data security and safeguard sensitive information in society, image encryption and decryption as well as pixel data modifications, are essential. To avoid misuse and preserve trust in our digital environment, it is crucial to use these technologies responsibly and ethically. So, to overcome some of the issues, the authors designed a way to modify pixel data that would hold the hidden information. The objective of this work is to change the pixel values in a way that can be used to store information about black and white image pixel data. Prior to encryption and decryption, by using Python we were able to construct a passcode with hand gestures in the air, then encrypt it without any data loss. It concentrates on keeping track of simply two pixel values. Thus, pixel values are slightly changed to ensure the masked image is not misleading. Considering that the RGB values are at their border values of 254, 255 the test cases of masking overcome issues with the corner values susceptibility

Imaging spectrum of extrapulmonary manifestations of COVID-19 infection- a multi-centre descriptive study from Southern India

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is the causative agent for coronavirus disease 2019 (COVID-19) pandemic. While the primary organ of involvement in this disease is the lung, multiple other organ systems can be involved either due to direct viral cytopathic effects or due to thrombo-inflammation and immune system dysregulation. In this study we describe the spectrum of extrapulmonary imaging findings encountered in our patients with COVID-19.Methods: This was a retrospective observational study conducted in three tertiary care hospitals in the city of Chennai in southern India. All cross-sectional imaging studies (other than lung imaging studies) performed in patients who had proven COVID-19 infection by RT-PCR testing during the period from April 2020 to March 2021 were included as part of the study. Extrapulmonary findings in these imaging studies were recorded and collated system-wise.Results: A total of 96 non-lung imaging studies were performed in patients who had RT-PCR positivity for COVID-19 infection. Among these a total of 30 studies had extrapulmonary imaging findings. Vascular involvement was seen in 14 patients, central nervous system involvement in 13 patients, abdomen involvement in 2 patients, and cardiac involvement in 1 patient. Vascular manifestations included arterial and venous thrombosis. Neurological manifestations included stroke, encephalitis and demyelination. Abdominal manifestations included enteritis and acute kidney injury. Cardiac manifestation was in form of myocarditis.Conclusions: Extrapulmonary imaging findings in COVID-19 are uncommon but not rare. Multisystem thrombotic manifestations and central nervous system involvement account for majority of extrapulmonary imaging findings in COVID-19

Delineating the structural, functional and evolutionary relationships of sucrose phosphate synthase gene family II in wheat and related grasses

<p>Abstract</p> <p>Background</p> <p>Sucrose phosphate synthase (SPS) is an important component of the plant sucrose biosynthesis pathway. In the monocotyledonous Poaceae, five <it>SPS </it>genes have been identified. Here we present a detailed analysis of the wheat <it>SPSII </it>family in wheat. A set of homoeologue-specific primers was developed in order to permit both the detection of sequence variation, and the dissection of the individual contribution of each homoeologue to the global expression of <it>SPSII</it>.</p> <p>Results</p> <p>The expression in bread wheat over the course of development of various sucrose biosynthesis genes monitored on an Affymetrix array showed that the <it>SPS </it>genes were regulated over time and space. <it>SPSII </it>homoeologue-specific assays were used to show that the three homoeologues contributed differentially to the global expression of <it>SPSII</it>. Genetic mapping placed the set of homoeoloci on the short arms of the homoeologous group 3 chromosomes. A resequencing of the A and B genome copies allowed the detection of four haplotypes at each locus. The 3B copy includes an unspliced intron. A comparison of the sequences of the wheat <it>SPSII </it>orthologues present in the diploid progenitors einkorn, goatgrass and <it>Triticum speltoides</it>, as well as in the more distantly related species barley, rice, sorghum and purple false brome demonstrated that intronic sequence was less well conserved than exonic. Comparative sequence and phylogenetic analysis of <it>SPSII </it>gene showed that false purple brome was more similar to <it>Triticeae </it>than to rice. Wheat - rice synteny was found to be perturbed at the SPS region.</p> <p>Conclusion</p> <p>The homoeologue-specific assays will be suitable to derive associations between SPS functionality and key phenotypic traits. The amplicon sequences derived from the homoeologue-specific primers are informative regarding the evolution of <it>SPSII </it>in a polyploid context.</p

Sustainable optimizing WMN performance through meta-heuristic TDMA link scheduling and routing

Wireless mesh networks (WMNs) have become a popular solution for expanding internet service and communication in both urban and rural areas. However, the performance of WMNs depends on generating optimized time-division multiple access (TDMA) schedules, which distribute time into a list of slots called superframes. This study proposes novel meta-heuristic algorithms to generate TDMA link schedules in WMNs using two different interference/constraint models: multi-transmit-receive (MTR) and full-duplex (FD). The objectives of this study are to optimize the TDMA frame for packet transmission, satisfy the constraints, and minimize the end-to-end delay. The significant contributions of this study are: (1) proposing effective and efficient heuristic solutions to solve the NP-complete problem of generating optimal TDMA link schedules in WMNs; (2) investigating the new FD interference model to improve the network capacity above the physical layer. To achieve these objectives and contributions, the study uses two popular meta-heuristics, the artificial bee colony (ABC) and/or genetic algorithm (GA), to solve the known NP-complete problems of joint scheduling, power control, and rate control. The results of this study show that the proposed algorithms can generate optimized TDMA link schedules for both MTR and FD models. The joint routing and scheduling approach further minimizes end-to-end delay while maintaining the schedule's minimum length and/or maximum capacity. The proposed solution outperforms the existing solutions in terms of the number of active links, end-to-end delay, and network capacity. The research aims to improve the efficiency and effectiveness of WMNs in most applications that require high throughput and fast response time

Multiple RSV strains infecting HEp-2 and A549 cells reveal cell line-dependent differences in resistance to RSV infection

Background: Respiratory syncytial virus (RSV) is the major viral driver of a global pediatric respiratory disease burden disproportionately borne by the poor1. Thus, RSV, like SARS-CoV-2, combines with congenital and environmental and host-history-dependent factors to create a spectrum of disease with greatest severity most frequently occurring in those least able to procure treatment. Methods: Here we apply whole genome sequencing and a suite of other molecular biological techniques to survey host-virus dynamics in infections of two distinct cell lines (HEp2 and A549) with four strains representative of known RSV genetic diversity. Results: We observed non-gradient patterns of RSV gene expression and a single major difference in transcriptional readthrough correlating with a deep split in the RSV phylogenetic tree. We also observed increased viral replication in HEp2 cells along with a pro-inflammatory host-response; and decreased viral replication in A549 cells with a more potent antiviral response in host gene expression and levels of secreted cytokines. Conclusions: Our findings suggest HEp2 and A549 cell lines can be used as complementary models of host response leading to more or less severe RSV disease. In vitro perturbations inspired by actual environmental and host-history-dependent factors associated with greater disease can be tested for their ability to shift the antiviral response of A549 cells to the more pro-inflammatory response of HEp2 cells. Such studies would help illuminate the tragic costs of poverty and suggest public health-level interventions to reduce the global disease burden from RSV and other respiratory viruses

Pan-Cancer Analysis of lncRNA Regulation Supports Their Targeting of Cancer Genes in Each Tumor Context

Long noncoding RNAs (lncRNAs) are commonly dys-regulated in tumors, but only a handful are known toplay pathophysiological roles in cancer. We inferredlncRNAs that dysregulate cancer pathways, onco-genes, and tumor suppressors (cancer genes) bymodeling their effects on the activity of transcriptionfactors, RNA-binding proteins, and microRNAs in5,185 TCGA tumors and 1,019 ENCODE assays.Our predictions included hundreds of candidateonco- and tumor-suppressor lncRNAs (cancerlncRNAs) whose somatic alterations account for thedysregulation of dozens of cancer genes and path-ways in each of 14 tumor contexts. To demonstrateproof of concept, we showed that perturbations tar-geting OIP5-AS1 (an inferred tumor suppressor) andTUG1 and WT1-AS (inferred onco-lncRNAs) dysre-gulated cancer genes and altered proliferation ofbreast and gynecologic cancer cells. Our analysis in-dicates that, although most lncRNAs are dysregu-lated in a tumor-specific manner, some, includingOIP5-AS1, TUG1, NEAT1, MEG3, and TSIX, synergis-tically dysregulate cancer pathways in multiple tumorcontexts

Pan-cancer Alterations of the MYC Oncogene and Its Proximal Network across the Cancer Genome Atlas

Although theMYConcogene has been implicated incancer, a systematic assessment of alterations ofMYC, related transcription factors, and co-regulatoryproteins, forming the proximal MYC network (PMN),across human cancers is lacking. Using computa-tional approaches, we define genomic and proteo-mic features associated with MYC and the PMNacross the 33 cancers of The Cancer Genome Atlas.Pan-cancer, 28% of all samples had at least one ofthe MYC paralogs amplified. In contrast, the MYCantagonists MGA and MNT were the most frequentlymutated or deleted members, proposing a roleas tumor suppressors.MYCalterations were mutu-ally exclusive withPIK3CA,PTEN,APC,orBRAFalterations, suggesting that MYC is a distinct onco-genic driver. Expression analysis revealed MYC-associated pathways in tumor subtypes, such asimmune response and growth factor signaling; chro-matin, translation, and DNA replication/repair wereconserved pan-cancer. This analysis reveals insightsinto MYC biology and is a reference for biomarkersand therapeutics for cancers with alterations ofMYC or the PMN