Reverse Protocol Research Articles

Mapping and quantification of potato virus A RNA genomes within viral particles and polysomes in infected plant cells.

Potato virus A belongs to the genus Potyvirus, a group of single-stranded positive sense RNA viruses infecting crops worldwide. To initiate infection in a host, its genome takes part in different activities, viz., translation, replication, encapsidation during the infection cycle. Extensive research has been carried out to scrutinize the stages of potyviral infection cycle and decipher the strategies it employs to cause disease. Nonetheless, the amount of viral RNA taking part in translation and virion formation, at a given time point, is missing. In this study, we quantified the percentage of viral RNA that exists as virions and those that associates with host polysome, relative to total viral RNA in infected plant tissue. We employed a revised version of immune-capture reverse transcription PCR and polysome profiling to address our queries. We tested three different coating antibody concentrations and further optimized the immuno-capture reverse transcription PCR protocol to address its limitation of binding and retaining viral particles. Our results indicate that most of the viral RNA (69%) exists as encapsidated genomes, while 3% of total viral RNA associates with host polysomes. These findings are crucial for correct interpretation of quantitative translational studies in which correlation must be made between the number of polysome-associated transcripts and the amount of protein synthesized.

Profiling of 24-nt phasiRNAs in Rice Anther by Massively Parallel Sequencing and Stem-Loop Reverse Transcription PCR.

Small RNAs are highly abundant and play important roles in plant reproduction. Profiling of small RNAs in reproductive tissues is a critical step in understanding their biology. Here, we describe a protocol for small RNA profiling in rice anthers, with a focus on an abundantly expressed but little-understood reproductive small RNA class named 24-nucleotide phased secondary small interfering RNAs (24-nt phasiRNAs). The protocol details small RNA library preparation steps for high-throughput sequencing, with a subsequent bioinformatic analysis framework. A low-throughput stem-loop reverse transcription PCR protocol for quick semi-quantification of small RNAs is also included as a complementary approach. The collective protocol, although focused on 24-nt phasiRNAs, should be amenable to other small RNA classes in various plant reproductive tissues.

Colombian cyanobacteria with cytotoxic activity in cancer cell lines

In the last decade, cyanobacteria have emerged as significant reservoirs of bioactive molecule for the pharmaceutical, cosmetic, and alimentary industries, given the wide spectrum of new possibilities of different organic pigments, proteins, carbohydrates, lipids, and powerful antioxidant sources for specific and stronger cancer treatments, autoimmune syndromes, obesity, and inflammatory diseases. A bioactivity screening was executed for 12 strains of Colombian cyanobacteria (10 marine and 2 freshwater) representative of the orders present in the LAUN culture collection, performing methanol extracts per sample, which was fractionated by reverse phase HPLC protocol, obtaining 8 fractions for each crude extract. All these crude extracts were tested for antimicrobial activities through disk diffusion methodology, and fractions were tested for cytotoxic activity against cancer cell lines by cell viability detection with MTT. Cyanobacteria's extracts showed considerable cytotoxic activity for osteosarcoma (MG063) and colon cancer (HCT116) cell lines (61 and 66 % of reduced viability compared to untreated controls, respectively) and a surprising cell growth promotion for the control group fibroblast (3T3L1) and brain endothelial (HCMEC) cell lines (121 and 127 % growth, respectively), no bioactivity was observed in the antimicrobial tests. These findings underscore the expansive cytotoxic potential of Colombian cyanobacteria against cancer cell lines and, notably, their growth-promoting effects on healthy cell lines. This positions them as promising sources of bioactive compounds for future pharmaceutical developments.

Learning competence of the temperate facultative cleaner wrasse Thalassoma pavo (Labridae).

Cleaner fishes remove parasites from other fishes called "clients," thereby contributing to a healthier ecosystem. Although the behavior and learning abilities of dedicated and tropical cleaner fishes have been broadly studied, a limited number of studies investigated the behavior of facultative and temperate cleaner fishes and, to the best of our knowledge, none focused on their cognitive abilities. Here, we tested the learning abilities of a species of temperate facultative cleaner, the ornate wrasse Thalassoma pavo in laboratory conditions, based on two problems. These two problems, or tasks, are relevant in two different contexts: the first one, deemed as mutualistically relevant, the cue-based discrimination task, and the second one, the spatial-based discrimination task, which is relevant in a non-cleaning context, when fish navigate through their environment to find food and return to their territories. We found that T. pavo individuals were able to solve these two tasks but excelled at the spatial task rather than with the cue discrimination. The same individuals were also challenged to learn the reverse protocol of these tasks and were again most successful in learning the reverse spatial discrimination problem, but not the cue. Contrary to the dedicated cleaner wrasse Labroides dimidiatus, this temperate facultative cleaner wrasse is slower to learn mutualistic relevant problems but competent in solving spatially derived problems. This may be due to the specific demands of their socio-ecological environment, with facultative cleaners having a greater component of non-mutualistic skills (spatial component), which prepares them to search for alternative food sources if necessary (e.g., feeding on the substrate) or even to expand territories more easily and less prepared to deal with mutualistic exchanges compared to dedicated cleaners that specialize to become increasingly socially competent.

Reverse scan body: Complete digital workflow for double full-arch zirconia prostheses.

During intraoral complete-arch digital implant scanning, one of the most technique-sensitive steps is the implant data acquisition and merging of different surface geometry data sets for prototype prosthesis fabrication due to the absence of fixed landmarks. Recently developed extraoral techniques have allowed for an alternative approach for the digital acquisition of implant position in completely edentulous patients. Specifically, extraoral digital scanning of the conversion prosthesis after connecting reverse scan bodies has been proposed as an efficient approach. The reverse scan body protocol digitally simulates the traditional back-pouring technique long utilized in analog workflows. The present article describes a technique for simplifying the digital workflow for the fabrication of passive-fitting definitive prostheses using the reverse scan body concept.

Combining Crystallization-Driven Self-Assembly with Reverse Sequence Polymerization-Induced Self-Assembly Enables the Efficient Synthesis of Hydrolytically Degradable Anisotropic Block Copolymer Nano-objects Directly in Concentrated Aqueous Media.

Herein we combine the well-known processing advantages conferred by polymerization-induced self-assembly (PISA) with crystallization-driven self-assembly (CDSA) to achieve the efficient synthesis of hydrolytically degradable, highly anisotropic block copolymer nano-objects directly in aqueous solution at 30% w/w solids. This new strategy involves a so-called reverse sequence PISA protocol that employs poly(l-lactide) (PLLA) as the crystallizable core-forming block and poly(N,N'-dimethylacrylamide) (PDMAC) as the water-soluble non-ionic coronal block. Such syntheses result in PDMAC-rich anisotropic nanoparticles. Depending on the target diblock copolymer composition, either rod-like nanoparticles or diamond-like platelets can be obtained. Furthermore, N-Acryloylmorpholine is briefly evaluated as an alternative hydrophilic vinyl monomer to DMAC. Given that the PLLA block can undergo either hydrolytic or enzymatic degradation, such nanoparticles are expected to offer potential applications in various fields, including next-generation sustainable Pickering emulsifiers.

Applications of Surface Plasmon Resonance (SPR) to the Study of Diverse Protein-Ligand Interactions.

Functional characterization of enzymes/proteins requires determination of the binding affinity of small molecules or other biomolecules with the target proteins. Several available techniques, such as proteomics and drug discovery strategies, require a precise and high-throughput assay for rapid and reliable screening of potential candidates for further testing. Surface plasmon resonance (SPR), a well-established label-free technique, directly measures biomolecular affinities. SPR assays require immobilization of one interacting component (ligand) on a conductive metal (mostly gold or silver) and a continuous flow of solution containing potential binding partner (analyte) across the surface. The SPR phenomenon occurs when polarized light excites the electrons at the interface of the metal and the dielectric medium to generate electromagnetic waves that propagate parallel to the surface. Changes in the refractive index due to interaction between the ligand and analyte are measured by detecting the reflected light, providing real-time data on kinetics and specificity. A prominent use of SPR is identifying compounds in crude plant extracts that bind to specific molecules. Procedures that utilize SPR are becoming increasingly applicable outside the laboratory setting, and SPR imaging and localized SPR (LSPR) are cheaper and more portable alternative for in situ detection of plant or mammalian pathogens and drug discovery studies. LSPR, in particular, has the advantage of direct attachment to test tissues in live-plant studies. Here, we describe three protocols utilizing SPR-based assays for precise analysis of protein-ligand interactions. © 2024 Wiley Periodicals LLC. Basic Protocol 1: SPR comparison of binding affinities of viral reverse transcriptase polymorphisms Basic Protocol 2: SPR screening of crude plant extract for protein-binding agents Basic Protocol 3: Localized SPR-based antigen detection using antibody-conjugated gold nanoparticles.

A Multiplex RT-PCR for the Detection of Three Viruses and One Viroid Infecting Hemp

Hemp ( Cannabis sativa L.) is an emerging industrial crop in the United States. In recent surveys of hemp growers, disease identification and management were determined to be significant priority areas for future research. From previous studies on disease identification in Western U.S. hemp production systems, hop latent viroid (HLVd), beet curly top virus (BCTV), Cannabis sativa mitovirus 1 (CasaMV1), and citrus yellow vein associated virus (CYVaV) were detected at high incidences. In the present study, a multiplex reverse transcription PCR protocol was developed to detect this viroid and viruses from hemp leaf samples simultaneously. Using the developed protocol, the multiplex reverse transcription PCR could detect each virus and viroid specifically in 10−1-diluted cDNA. The protocol was validated on 6 hemp metavirome leaf samples from field-grown hemp grown in 2019 and 23 uncharacterized leaf samples in 2021 from Colorado. Of the samples tested, 95% tested positive for CasaMV1 in 2021. This was greater than infections of BCTV (4%), HLVd (0%), and CYVaV (0%). Coinfections accounted for 4% of samples in 2021. This method offers sensitive, specific, and fast simultaneous detection of three viruses and one viroid from hemp, contributing to large-scale pathogen-free hemp certification schemes. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Reverse Genetics of Murine Rotavirus: A Comparative Analysis of the Wild-Type and Cell-Culture-Adapted Murine Rotavirus VP4 in Replication and Virulence in Neonatal Mice.

Small-animal models and reverse genetics systems are powerful tools for investigating the molecular mechanisms underlying viral replication, virulence, and interaction with the host immune response in vivo. Rotavirus (RV) causes acute gastroenteritis in many young animals and infants worldwide. Murine RV replicates efficiently in the intestines of inoculated suckling pups, causing diarrhea, and spreads efficiently to uninoculated littermates. Because RVs derived from human and other non-mouse animal species do not replicate efficiently in mice, murine RVs are uniquely useful in probing the viral and host determinants of efficient replication and pathogenesis in a species-matched mouse model. Previously, we established an optimized reverse genetics protocol for RV and successfully generated a murine-like RV rD6/2-2g strain that replicates well in both cultured cell lines and in the intestines of inoculated pups. However, rD6/2-2g possesses three out of eleven gene segments derived from simian RV strains, and these three heterologous segments may attenuate viral pathogenicity in vivo. Here, we rescued the first recombinant RV with all 11 gene segments of murine RV origin. Using this virus as a genetic background, we generated a panel of recombinant murine RVs with either N-terminal VP8* or C-terminal VP5* regions chimerized between a cell-culture-adapted murine ETD strain and a non-tissue-culture-adapted murine EW strain and compared the diarrhea rate and fecal RV shedding in pups. The recombinant viruses with VP5* domains derived from the murine EW strain showed slightly more fecal shedding than those with VP5* domains from the ETD strain. The newly characterized full-genome murine RV will be a useful tool for dissecting virus-host interactions and for studying the mechanism of pathogenesis in neonatal mice.

Epidemiologic Survey of Crimean-Congo Hemorrhagic Fever Virus in Suids, Spain.

We conducted a cross-sectional study in wild boar and extensively managed Iberian pig populations in a hotspot area of Crimean-Congo hemorrhagic fever virus (CCHFV) in Spain. We tested for antibodies against CCHFV by using 2 ELISAs in parallel. We assessed the presence of CCHFV RNA by means of reverse transcription quantitative PCR protocol, which detects all genotypes. A total of 113 (21.8%) of 518 suids sampled showed antibodies against CCHFV by ELISA. By species, 106 (39.7%) of 267 wild boars and 7 (2.8%) of 251 Iberian pigs analyzed were seropositive. Of the 231 Iberian pigs and 231 wild boars analyzed, none tested positive for CCHFV RNA. These findings indicate high CCHFV exposure in wild boar populations in endemic areas and confirm the susceptibility of extensively reared pigs to CCHFV, even though they may only play a limited role in the enzootic cycle.

An Optimized Mouse Embryonic Stem Cell Based Reverse Poly-Transfection Technique for Rapid Exploration of Nucleic Acid Ratios.

Due to its relative simplicity and ease of use, transient transfection of mammalian cell lines with nucleic acids has become a mainstay in biomedical research. While most widely used cell lines have robust protocols for transfection in adherent two-dimensional culture, these protocols often do not translate well to less-studied lines or those with atypical, hard-to-transfect morphologies. Using mouse pluripotent stem cells grown in 2i/LIF media, a widely used culture model for regenerative medicine, this method outlines an optimized, rapid reverse transfection protocol capable of achieving higher transfection efficiency. Leveraging this protocol, a three-plasmid poly-transfection is performed, taking advantage of the higher-than-normal efficiency in plasmid delivery to study an expanded range of plasmid stoichiometry. This reverse poly-transfection protocol allows for a one-pot experimental method, enabling users to optimize plasmid ratios in a single well, rather than across several co-transfections. By facilitating the rapid exploration of the effect of DNA stoichiometry on the overall function of delivered genetic circuits, this protocol minimizes the time and cost of embryonic stem cell transfection.

Seismic performance evaluation of plastered cellular lightweight concrete (CLC) block masonry walls

The current research presents a novel and sustainable load-bearing system utilizing cellular lightweight concrete block masonry walls. These blocks, known for their eco-friendly properties and increasing popularity in the construction industry, have been studied extensively for their physical and mechanical characteristics. However, this study aims to expand upon previous research by examining the seismic performance of these walls in a seismically active region, where cellular lightweight concrete block usage is emerging. The study includes the construction and testing of multiple masonry prisms, wallets, and full-scale walls using a quasi-static reverse cyclic loading protocol. The behavior of the walls is analyzed and compared in terms of various parameters such as force–deformation curve, energy dissipation, stiffness degradation, deformation ductility factor, response modification factor, and seismic performance levels, as well as rocking, in-plane sliding, and out-of-plane movement. The results indicate that the use of confining elements significantly improves the lateral load capacity, elastic stiffness, and displacement ductility factor of the confined masonry wall in comparison to an unreinforced masonry wall by 102%, 66.67%, and 5.3%, respectively. Overall, the study concludes that the inclusion of confining elements enhances the seismic performance of the confined masonry wall under lateral loading.

Screening of camphene as a potential inhibitor targeting SARS-CoV-2 various structural and functional mutants: Through reverse docking approach

Background: SARS-CoV was first identified in 2003 but SARS-CoV-2, which gained its recognition again in 2019 as COVID-19, has been a crucial threat worldwide and has caused more death rates than the SARS-CoV but till now no confined treatments are available. The present study aimed to investigate the efficacy of camphene against various structural and functional mutants of SARS-CoV-2 using reverse docking protocol. Methods: To investigate the efficacy of camphene as a potential antiviral drug against COVID-19, against of all possible target proteins in SARS-CoV-2, which could lead to a new platform for drug discovery. Reverse pharmacology (Reverse docking) approach was performed, which involved docking of camphene and 20 structural and non-structural proteins (NSPs) of SARS-CoV-2 performed using maestro 12.8 of Schrödinger. Results: The results were evaluated since the minimum binding energy obtained after docking and camphene was effective against most of the proteins responsible for SARS-CoV-2, but camphene showed greater efficacy against the main protease (protease 9), which is main functional protein of SARS-CoV-2. Hence, the study proves that camphene can be a good drug candidate for different mutants of SARS-CoV-2. Conclusion: Protease 9, which is the main functional protein of SARS-CoV-2, expressed the best binding affinity with camphene having the minimum binding energy (-5.616). Hence, it is concluded that camphene could be the drug contender against protease 9 as it is a more potent target in SARS-CoV-2. This could be a major finding, as camphene is related to camphor, which is already very beneficial against many respiratory problems.

Medial and lateral dual plating of native distal femur fractures: a systematic literature review.

Lateral locked plating (LLP) development has improved outcomes for distal femur fractures. However, there is still a modest rate of nonunion in fractures treated with LLP alone, with higher nonunion risk in high-energy fractures, intra-articular involvement, poor bone quality, severe comminution, or bone loss. Several recent studies have demonstrated both the safety and the biomechanical advantage of dual medial and lateral plating (DP). The purpose of this study was to evaluate the clinical outcomes of DP for native distal femoral fractures by performing a systematic review of the literature. Studies reporting clinical outcomes for DP of native distal femur fractures were identified and systematically reviewed. Publications without full-text manuscripts, those solely involving periprosthetic fractures, or fractures other than distal femur fractures were excluded. Fracture type, mean follow-up, open versus closed fracture, number of bone grafting procedures, nonunion, reoperation rates, and complication data were collected. Methodologic study quality was assessed using the Coleman methodology score. The initial electronic review and reverse inclusion protocol identified 1484 publications. After removal of duplicates and abstract review to exclude studies that did not discuss clinical treatment of femur fractures with dual plating, 101 potential manuscripts were identified and manually reviewed. After final review, 12 studies were included in this study. There were 199 fractures with average follow-up time of 13.72 months. Unplanned reoperations and nonunion occurred in 19 (8.5%) and 9 (4.5%) cases, respectively. The most frequently reported complications were superficial infection (n = 6, 3%) and deep infection (n = 5, 2.5%) postoperatively. Other complications included delayed union (n = 6, 3%) not requiring additional surgical treatment and knee stiffness in four patients (2%) necessitating manipulation under anesthesia or lysis of adhesions. The average Coleman score was 50.5 (range 13.5-72), suggesting that included studies were of moderate-to-poor quality. Clinical research interest in DP of distal femoral fractures has markedly increased in the past few decades. The current data suggest that DP of native distal femoral fractures is associated with favorable nonunion and reoperation rates compared with previously published rates associated with LLP alone. In the current review, DP of distal femoral fractures was associated with acceptable rates of complications and generally good functional outcomes. More high-quality, directly comparable research is necessary to validate the conclusions of this review.

Clinical Practice Guidelines for the Use of Transcranial Direct Current Stimulation in Psychiatry.

Clinical Practice Guidelines for the Use of Transcranial Direct Current Stimulation in Psychiatry.

Modeling and analysis of traffic warning message dissemination system in VANETs

As the preferred underpinning technology of intelligent transportation systems (ITS), vehicular ad hoc networks (VANETs) have been widely used to cope with various traffic problems. However, implementing application-specific message dissemination in VANETs with fewer message dissemination is a challenge since, in fact, it is not meaningful for some vehicles to get such a message for the application requirements for alleviating traffic congestion. Thus, this paper proposes a traffic warning message dissemination system (TWMDS) framework together with a protocol for traffic warning messages (TWMs) dissemination called the reverse routing protocol (RRP). With TWMDS, when an abnormal traffic event (ATE) occurs, the vehicle causing this event generates a standardized TWM. This message is then forwarded only to the vehicles that may enter the potential congestion area (PCA) via vehicle-to-vehicle and vehicle-to-intersection interactions. In addition, three colored petri net models are developed to model and analyze the interaction behavior of TWMDS. In this way, we show the correctness and reliability of RRP and provide a feasibility measure for the subsequent development and implementation of TWMDS. Finally, we use Veins framework integrating OMNeT++ and SUMO to simulate the TWMDS functional modules, which further validates that RRP has a higher application value compared with three other routing protocols in TWMDS. With TWMDS, drivers can re-route their traveling paths as soon as possible to alleviate traffic congestion.

Reverse auction protocol based on quantum teleportation

Reverse auction is a revolutionary procurement technology, widely used in enterprises and governments, which can increase purchasing transparency and reduce purchasing costs. In this paper, we combine reverse auction with quantum theory and propose a novel protocol that guarantees security by adopting quantum teleportation, quantum key distribution and so on. This protocol also has the properties of anonymity of supplier, verifiability, disavowal impossibility and traceability, properties that ensure the fairness of transactions. The analysis results show that the protocol is simple, high performing and reliable. We hope that it will be applied in real life.

A Real-World Population Pharmacokinetic (PK) Study of Two Recombinant Factor VIII (rFVIII) Concentrates in Boys with Severe Hemophilia a without Inhibitors

Introduction To personalize long-term prophylaxis in boys with hemophilia A with a severe bleeding phenotype, the individual's pharmacokinetic (PK) profile must be established. Traditional FVIII PK assessment has required a 2-3-day washout, which can place boys at risk of bleeding, and up to 11 post-infusion blood samples collected over a 48h period (Lee et al., ISTH guidelines, 2001). Population PK models with Bayesian forecasting are currently available that permit the use of a no-washout PK protocol with sparse sampling, which is clinically practical and less demanding for patients and their families. Our previous experience with a no-washout, single-clinic visit, reverse 2-sampling time point (24 and 3h) PK protocol established that this practical, patient-friendly PK protocol was sufficiently accurate for generating clinically useful individual PK profiles in patients receiving a standard half-life (SHL) FVIII concentrate [antihemophilic factor (recombinant) plasma/albumin-free method (Advate®)] for long-term prophylaxis (Blanchette et al., Thromb Haemost, 2021). Aim To determine the accuracy of a clinically practical single-clinic visit, reverse 2-sampling time point population PK protocol for generating individual PK profiles [clearance (Cl) and terminal half-life (t1/2)] in boys with severe hemophilia A without inhibitors receiving either an SHL [antihemophilic factor (recombinant) BAY 81-8973 (Kovaltry®)] or an extended half-life (EHL) FVIII concentrate [antihemophilic factor (recombinant) PEGylated BAY 94-9027 (Jivi®)] for long-term prophylaxis. Methods In this multicenter observational study, participants had their first infusion given at home. Of a total of 14 boys with hemophilia A without inhibitors receiving long-term prophylaxis, 5 receiving a SHL completed a single clinic visit, reverse 2-point (24 and 3h) and a multiple clinic visit 6-point (baseline, 1, 3, 8, 24 and 48h) PK sampling protocol; and 4 boys completed a SHL single clinic visit reverse 2-point (48 and 3h) and a 2 clinic visit reverse 3-point (48, 3 and 24h) PK sampling protocol. 5 boys receiving EHL completed single clinic visit, reverse 2-point (72 and 3h) and 2-clinic visit 3-point (72, 3 and 48h) PK sampling protocols and they had multiple spot samplings (n=12) at various post-infusion times to evaluate the precision of their predicted PK profiles based on their 2-point PK sampling protocol. One-stage (OS) and chromogenic (CHR) FVIII:C and von Willebrand factor antigen (VWF:Ag) assays were performed in a central reference coagulation laboratory in Kingston, Ontario. PK parameters (Cl and t1/2) were estimated using the WAPPS-Hemo population PK program. The median of the % difference between the 2-sampling time point PK protocol compared to the 6- and 3- sampling time point PK protocols is reported. Results 14 boys with hemophilia A (FVIII:C <2%; median age: 14.5 yrs, range: 5-19 yrs) participated in the study. The 2-point PK protocol for both SHL and EHL showed a higher Cl and lower t1/2 when compared to either a 6-sampling time point (for SHL) or a 3-sampling time point (for either SHL or EHL) PK assessment (Table 1). The difference was greatest for the 2-point (24 and 3h) vs the 6-point (Table 1). In the case of the 2-point (48 and 3h for SHL and 72 and 3h for EHL) vs 3-point (an additional 24h time point for SHL and 48h for EHL) PK parameter estimates, the 2-point protocol gave values that were all within 6% of those obtained with the 3-sampling time point PK (Table 1). The OS and CHR assays gave similar estimations for Cl and t1/2 with the exception of t1/2 when comparing the 6-point to the 2-point PK protocol (Table 1). Evaluation of the 2-sampling time point PK protocol through spot sampling showed approximately 66% and 81% of FVIII:C values falling within the 95% prediction limits for the OS and CHR assays, respectively (n=12 for EHL OS; n=11 for EHL CHR). There was a strong association between the observed FVIII:C levels from the spot sampling and their predicted values from the 2-point PK for EHL (R2=94% and 71% for the OS and CHR assays, respectively) (Fig. 1). Conclusion The reverse 2-point, single clinic visit, PK protocol (24/48 or 72h and 3h) is a practical approach to generate PK parameters that provide clinically useful information for guiding personalized prophylaxis regimens for boys receiving SHL or EHL for long-term prophylaxis. Funding Supported by Bayer (Investigator-Initiated Research grant) Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

CETS: Enabling Sustainable IoT with Cooperative Energy Transfer Schedule towards 6G Era.

The large scale of the Internet of Things necessitates using long-lasting physical layer devices for data collection. Deploying large numbers of Wi-Fi-enabled devices is expensive, so the Internet of Everything (IoE) is equipped with multiple communication modules to collect data where Wi-Fi is unavailable. However, because of their extended communication capabilities, IoE devices face energy limitations. As a result, IoE devices must be provided with the necessary energy resources. This paper introduces a novel multi-hop cooperation communication mechanism for Wireless Energy Transfer (WET) in the Wireless Powered-Internet of Everything (WP-IoE). IoE devices are outfitted here with various communication devices such as RF, Bluetooth, and Wi-Fi. This research proposes a two-phase energy transmission schedule to address the energy requirements. For data collection, the first phase provides a distributed tree-based data communication plan. The proposed model’s second phase used the reverse data collection protocol to implement wireless energy transmission. By combining these two phases, an optimized WET framework was created without unmanned aerial vehicles or robots. The experimental findings show that the proposed method in this research increases the average lifetime of the network and has a more significant charge latency and average charge throughput than other models.

An Efficient Modern Strategy to Screen Drug Candidates Targeting RdRp of SARS-CoV-2 With Potentially High Selectivity and Specificity.

Desired drug candidates should have both a high potential binding chance and high specificity. Recently, many drug screening strategies have been developed to screen compounds with high possible binding chances or high binding affinity. However, there is still no good solution to detect whether those selected compounds possess high specificity. Here, we developed a reverse DFCNN (Dense Fully Connected Neural Network) and a reverse docking protocol to check a given compound’s ability to bind diversified targets and estimate its specificity with homemade formulas. We used the RNA-dependent RNA polymerase (RdRp) target as a proof-of-concept example to identify drug candidates with high selectivity and high specificity. We first used a previously developed hybrid screening method to find drug candidates from an 8888-size compound database. The hybrid screening method takes advantage of the deep learning-based method, traditional molecular docking, molecular dynamics simulation, and binding free energy calculated by metadynamics, which should be powerful in selecting high binding affinity candidates. Also, we integrated the reverse DFCNN and reversed docking against a diversified 102 proteins to the pipeline for assessing the specificity of those selected candidates, and finally got compounds that have both predicted selectivity and specificity. Among the eight selected candidates, Platycodin D and Tubeimoside III were confirmed to effectively inhibit SARS-CoV-2 replication in vitro with EC50 values of 619.5 and 265.5 nM, respectively. Our study discovered that Tubeimoside III could inhibit SARS-CoV-2 replication potently for the first time. Furthermore, the underlying mechanisms of Platycodin D and Tubeimoside III inhibiting SARS-CoV-2 are highly possible by blocking the RdRp cavity according to our screening procedure. In addition, the careful analysis predicted common critical residues involved in the binding with active inhibitors Platycodin D and Tubeimoside III, Azithromycin, and Pralatrexate, which hopefully promote the development of non-covalent binding inhibitors against RdRp.