Gossypium hirsutum calmodulin-like protein (CML 11) interaction with geminivirus encoded protein using bioinformatics and molecular techniques.

Plant viruses are the most abundant destructive agents that exist in every ecosystem, causing severe diseases in multiple crops worldwide. Currently, a major gap is present in computational biology determining plant viruses interaction with its host. We lay out a strategy to extract virus-host protein interactions using various protein binding and interface methods for Geminiviridae, a second largest virus family. Using this approach, transcriptional activator protein (TrAP/C2) encoded by Cotton leaf curl Kokhran virus (CLCuKoV) and Cotton leaf curl Multan virus (CLCuMV) showed strong binding affinity with calmodulin-like (CML) protein of Gossypium hirsutum (Gh-CML11). Higher negative value for the change in Gibbs free energy between TrAP and Gh-CML11 indicated strong binding affinity. Consensus from gene ontology database and in-silico nuclear localization signal (NLS) tools identified subcellular localization of TrAP in the nucleus associated with Gh-CML11 for virus infection. Data based on interaction prediction and docking methods present evidences that full length and truncated C2 strongly binds with Gh-CML11. This computational data was further validated with molecular results collected from yeast two-hybrid, bimolecular fluorescence complementation system and pull down assay. In this work, we also show the outcomes of full length and truncated TrAP on plant machinery. This is a first extensive report to delineate a role of CML protein from cotton with begomoviruses encoded transcription activator protein.

Epidemiology of Brucellosis in Small Ruminants of Rural and Peri-Urban Areas of Multan, Pakistan.

Brucellosis is a widespread zoonotic disease of veterinary and public health importance with considerably higher prevalence in developing/underdeveloped countries. This study reports the prevalence and risk determinants of brucellosis in small ruminants of peri-urban and rural areas of district Multan, Southern Punjab, Pakistan. For this purpose, sera samples (n = 392) of small ruminants were collected and subjected to preliminary screening using commercially available RBPT reagents followed by serodetection of brucellosis using multispecies i-ELISA kit (ID.vet, France). All the ELISA positive samples were confirmed by PCR using genus-specific primers, and frequencies of Brucella species in positive samples were enumerated using species-specific primers. Results indicated seropositivity rates of 9.69, 9.95, and 10.20% in study population using RBPT reagents of IDEXX-USA, ID.Vet-France, and VRI-Pakistan, respectively, with a statistically nonsignificant difference (p > 0.05). Results of ELISA showed an overall seroprevalence rate of 7.14% in target population with a slightly higher rate in sheep (7.65%) as compared to goat (6.63%) population (p = 0.695; OR = 1.16, 95% CI = 0.53, 2.57). Results revealed that out of total positive samples, B. abortus was detected in 60.71% of seropositive samples and B. melitensis was detected in 14.28% of positive samples. It was revealed that risk factors including body condition scores, hygienic conditions of the housing facility, farming system, reproductive disorders, educational status of farmers, and awareness of farmers about brucellosis had significant association with brucellosis in small ruminants of study area (p < 0.05). Conversely, farm/herd size, locality, gender, age, weight, and parity showed a nonsignificant association (p > 0.05) with brucellosis. In conclusion, brucellosis is prevalent in small ruminants of Multan, Pakistan. It is recommended to devise and implement effective control strategies with a major focus on raising awareness about brucellosis in farmers for the containment of infection in the region.

A study on the epidemiology of brucellosis in bovine population of peri-urban and rural areas of district Multan, southern Punjab, Pakistan.

BACKGROUND: Brucellosis is a zoonotic disease caused by a bacterial pathogen belonging to the genus Brucella. It is one of the most frequent bacterial zoonoses globally but unfortunately, it is still considered as a neglected disease in the developing world. Keeping in view, this study was conducted to determine the prevalence and risk determinants of brucellosis in large ruminants of peri-urban and rural areas of district Multan-Pakistan. For this purpose, blood samples (n = 490) were collected from the cattle (n = 245) and buffalo (n = 245) population of the study area and subjected to preliminary screening of brucellosis using local and imported RBPT reagents. All the samples were further analyzed using commercially available multi-specie indirect ELISA kit followed by their confirmation by PCR using genus and species-specific primers. Data obtained from lab analysis and questionnaires were subjected to statistical analysis for Pearson Chi-square, Odds Ratio and Confidence intervals (95%). RESULTS: The results showed that the maximum seropositivity was recorded with local RBPT reagent (VRI, Pakistan; 12.45%; 95%CI = 9.72-15.65%) followed by RBPT-IDEXX (12.24%; 95%CI = 9.52-15.45%) and RBPT-ID.vet (11.84%; 95%CI = 9.18-14.95%) however statistical difference was non-significant (P = 0.956). The ELISA results showed an overall seroprevalence rate of 11.22% (95%CI = 8.59-14.33%) with comparatively higher rate in cattle (12.65%; 95%CI = 8.82-17.44%) as compared to buffaloes (9.80%; 95%CI = 6.49-14.15%). The PCR analysis confirmed the presence of genus Brucella in all seropositive samples whereas frequency of B. abortus and B. melitensis in seropositive samples was 80% and 20%, respectively. The co-existence of both species was also observed in 5.45% samples. The statistical analysis showed a significant association of bovine brucellosis with herd size, breed, reproductive disorders, mode of insemination, educational status and farmers' awareness about brucellosis (P < 0.05). Conversely, locality, age, weight, gender, pregnancy status, parity and puberty status had no associations with brucellosis (P > 0.05). CONCLUSION: In conclusion, brucellosis is prevalent in large ruminants of district Multan, Pakistan. It is suggested to devise and implement stringent policies for the effective control and prevention of brucellosis in the region. Further, the current situation also warrants the need to strengthen interdisciplinary coordination among veterinarians and physicians in one health perspective to ensure and strengthen the human and animal health care systems in the region.

Biotechnology and Solutions: Insect-Pest-Resistance Management for Improvement and Development of Bt Cotton (Gossypium hirsutum L.).

Cotton (Gossypium spp. L.) is a major origin of natural fiber, and is projected at 117 million bales worldwide for 2021/22. A variety of biotic and abiotic stresses have considerable negative impacts on cotton. The significantly decreased applications of chemical insecticidal sprays in the agro-ecosystem have greatly affected the biodiversity and dynamics of primary and secondary insects. Various control measures were taken around the globe to increase production costs. Temperature, drought, and salinity, and biotic stresses such as bacteria, viruses, fungi, nematodes, insects, and mites cause substantial losses to cotton crops. Here, we summarize a number of biotic and abiotic stresses upsetting Bt cotton crop with present and future biotechnology solution strategies that include a refuge strategy, multi-gene pyramiding, the release of sterile insects, seed mixing, RNAi, CRISPR/Cas9, biotic signaling, and the use of bioagents. Surveillance of insect resistance, monitoring of grower compliance, and implementation of remedial actions can lead to the sustainable use of cotton across the globe.

Drought and heat stress on cotton genotypes suggested agro-physiological and biochemical features for climate resilience.

This study aimed to investigate the impact of individual drought, heat, and combined drought and heat stress on twelve cotton genotypes, including eight tolerant and four susceptible genotypes. A field experiment was carried out by employing a randomized complete block split-plot design, with treatments (control, drought, heat, drought + heat), and cotton genotypes assigned to the main plots and sub-plots respectively. The results showed that the combined stress had a more severe impact on the yield and fiber quality of cotton genotypes compared to individual stresses. Among the studied genotypes, FB-Shaheen, FH-207, MNH-886, and White Gold exhibited superior performance in regard to agronomic and fiber quality characters under combined stress environments. Physiological parameters, including transpiration rate, stomatal conductance, relative water contents, and photosynthetic rate, were significantly reduced under combined stress. However, specific genotypes, MNH-886, FH-207, White Gold, and FB-Shaheen, demonstrated better maintenance of these parameters, indicating their enhanced tolerance to the combined stress. Furthermore, the accumulation of reactive oxygen species was more pronounced under combined stress compared to individual stressors. Tolerant genotypes showed lower levels of H2O2 and MDA accumulation, while susceptible genotypes exhibited higher levels of oxidative damage. Antioxidant enzyme activities, such as superoxide dismutase, peroxidase, and catalase, increased under combined stress, with tolerant genotypes displaying higher enzyme activities. Conversely, susceptible genotypes (AA-703, KZ 191, IR-6, and S-15) demonstrated lower increases in enzymatic activities under combined stress conditions. Biochemical traits, including proline, total phenolic content, flavonoids, and ascorbic acid, exhibited higher levels in resistant genotypes under combined stress, while sensitive genotypes displayed decreased levels of these traits. Additionally, chlorophyll a & b, and carotenoid levels were notably decreased under combined stress, with tolerant genotypes experiencing a lesser decrease compared to susceptible genotypes.

Prevalence of antibodies to Toxocara canis and its associated risk factors in socio-economically deprived nomadic communities of Pakistan.

Toxocariasis is an important zoonotic disease caused by Toxocara (T.) canis with considerably higher prevalence in developing countries. The data on its epidemiology, especially in socioeconomically deprived nomadic communities, are scarce in Pakistan. Therefore, this study was conducted to determine the prevalence of anti-T. canis antibodies and its associated risk factors in nomadic communities located in and around Multan, Pakistan. A total of 184 sera samples were collected from nomadic communities by simple random sampling technique. The descriptive epidemiological data of participants were collected on well-designed questionnaires. Prior consent was also obtained from the participants to use the data generated from their samples without showing their identity. All the samples were analysed for the detection of anti-T. canis antibodies using commercially available Enzyme-Linked-Immunosorbent-Assay (ELISA) kits having 91% sensitivity and 96% specificity (Bordier Affinity Products, Switzerland). The overall seroprevalence of toxocariasis among nomadic communities was 27.7% (51/184). Various factors, including age, known disease history, nutritional status, contact with dogs, practice of hand washing after contact with dogs, use of unwashed vegetables, body mass index, and drug abuse, showed significant correlation (p < 0.05) with toxocariasis in nomadic communities. Conversely, other factors, including gender, marital status, educational status, awareness about zoonotic diseases, source of drinking water, occupation, location, hand washing before taking food, exposure to soil, and hygienic eating behaviour, showed non-significant correlation (p > 0.05) with seroprevalence of toxocariasis. Results also showed that >50% of seropositive cases were asymptomatic, whereas cough and abdominal pain were recorded in 19.6% and 11.76% of seropositive cases, respectively. Keeping in view, it is suggested to conduct surveys at mass level to rule out the exact disease status at national level and to include nomadic communities in local, national, and regional disease control programs through provision of better healthcare facilities and awareness about the disease.

Breakthrough in CRISPR/Cas system: Current and future directions and challenges.

Targeted genome editing (GE) technology has brought a significant revolution in fictional genomic research and given hope to plant scientists to develop desirable varieties. This technology involves inducing site-specific DNA perturbations that can be repaired through DNA repair pathways. GE products currently include CRISPR-associated nuclease DNA breaks, prime editors generated DNA flaps, single nucleotide-modifications, transposases, and recombinases. The discovery of double-strand breaks, site-specific nucleases (SSNs), and repair mechanisms paved the way for targeted GE, and the first-generation GE tools, ZFNs and TALENs, were successfully utilized in plant GE. However, CRISPR-Cas has now become the preferred tool for GE due to its speed, reliability, and cost-effectiveness. Plant functional genomics has benefited significantly from the widespread use of CRISPR technology for advancements and developments. This review highlights the progress made in CRISPR technology, including multiplex editing, base editing (BE), and prime editing (PE), as well as the challenges and potential delivery mechanisms.

Blockchain in Healthcare: A Decentralized Platform for Digital Health Passport of COVID-19 Based on Vaccination and Immunity Certificates.

COVID-19 has become a very transmissible disease that has had a worldwide impact, resulting in a huge number of infections and fatalities. Testing is critical to the pandemic's successful response because it helps detect illnesses and so attenuate (isolate/cure) them and now vaccination is a life-safer innovation against the pandemic which helps to make the immunity system stronger and fight against this infection. Patient-sensitive information, on the other hand, is now held in a centralized or third-party storage paradigm, according to COVID-19. One of the most difficult aspects of using a centralized storage strategy is maintaining patient privacy and system transparency. The application of blockchain technology to support health initiatives that can minimize the spread of COVID-19 infections in the context of accessibility of the system and for verification of digital passports. Only by combining blockchain technology with advanced cryptographic algorithms can a secure and privacy-preserving solution to COVID-19 be provided. In this article, we investigate the issue and propose a blockchain-based solution incorporating conscience identity, encryption, and decentralized storage via interplanetary file systems (IPFS). For COVID-19 test takers and vaccination takers, our solution includes digital health passports (DHP) as a certification of test or vaccination. We explain smart contracts constructed and tested with Ethereum to preserve a DHP for test and vaccine takers, allowing for a prompt and trustworthy response from the necessary medical authorities. We use an immutable trustworthy blockchain to minimize medical facility response times, relieve the transmission of incorrect information, and stop the illness from spreading via DHP. We give a detailed explanation of the proposed solution's system model, development, and assessment in terms of cost and security. Finally, we put the suggested framework to the test by deploying a smart contract prototype on the Ethereum TESTNET network in a Windows environment. The study's findings revealed that the suggested method is effective and feasible.

Decentralized Patient-Centric Report and Medical Image Management System Based on Blockchain Technology and the Inter-Planetary File System.

Several academicians have been actively contributing to establishing a practical solution to storing and distributing medical images and test reports in the research domain of health care in recent years. Current procedures mainly rely on cloud-assisted centralized data centers, which raise maintenance expenditure, necessitate a large amount of storage space, and raise privacy concerns when exchanging data across a network. As a result, it is critically essential to provide a framework that allows for the efficient exchange and storage of large amounts of medical data in a secure setting. In this research, we describe a unique proof-of-concept architecture for a distributed patient-centric test report and image management (PCRIM) system that aims to facilitate patient privacy and control without the need for a centralized infrastructure. We used an Ethereum blockchain and a distributed file system technology called the Inter-Planetary File System in this system (IPFS). Then, to secure a distributed and trustworthy access control policy, we designed an Ethereum smart contract termed the patient-centric access control protocol. The IPFS allows for the decentralized storage of medical metadata, such as images, with worldwide accessibility. We demonstrate how the PCRIM system design enables hospitals, patients, and image requestors to obtain patient-centric data in a distributed and secure manner. Finally, we tested the proposed framework in the Windows environment by deploying a smart contract prototype on an Ethereum TESTNET blockchain. The findings of the study indicate that the proposed strategy is both efficient and practicable.

Impact of IL-21 Gene Polymorphisms (rs2055979) and the Levels of Serum IL-21 on the Risk of Multiple Sclerosis.

Multiple sclerosis (MS) is a chronic autoimmune disorder of the central nervous system. Genetic and environmental factors have important roles in the induction, onset, and progression of MS. In this study, the IL-21 genotype (rs2055979) (G/T) in Iraqi MS patients was compared with a healthy control group to investigate the possible association of any particular genotype or allele with multiple sclerosis. This study included 70 patients with relapsing-remitting MS and 50 healthy individuals as control. Following the extraction of genomic DNA, polymerase chain reaction-restriction fragment length polymorphism, the frequencies of genotypes, and alleles were calculated and statistically analyzed. The results of the study revealed a significant reduction in the distribution of the wild homozygous genotype (GG) in MS patients, in comparison to a healthy control group (14.3% vs. 34 %; 0.0129 at P≤0.05; odds ratio [OR] 3.0909, 95% confidence interval [CI]: 1.2704-7.5203). However, MS cases and controls did not differ significantly in neither GT nor TT genotypes, 62.9% (OR 0.6402, 95% CI: 0.3064-1.3374) and 52% (OR 0.5494, 95% CI: 0.2074-1.4557), respectively. The data of allele frequencies in patients and controls showed that the G allele frequencies were 0.46 vs. 0.60 in patients and controls, respectively, while T allele frequencies were 0.54 vs. 0.40 in patients and controls, respectively. The current conclusions indicated that in the study group, the GG genotype of IL-21(rs2055979) could be related to MS.

Heterologous expression of cry3Bb1 and cry3 genes for enhanced resistance against insect pests in cotton.

Transgenic technology played a crucial role in developing insect-resistant plants resulting in the reduced application of pesticides. This article reports the expression of two cry proteins (Cry3Bb1 and Cry3) in cotton for enhanced resistance against chewing insect pests. The aforementioned genes were synthetically developed and were cloned under appropriate regulatory sequences followed by transformation into Eagle-2 genotype (Gossypium hirsutum) of cotton through shoot apex-cut Agro-infiltration. The transgene integration was validated by polymerase chain reaction using primers flanking the aforementioned cry genes. Transgene expression was assessed by qRT-PCR using GADPH as a reference gene. The relative fold expression analyses revealed the highest expression of the transgene(s) in M1 plants, which is a 4.5-fold expression (Cry3 + Cry3Bb1) followed by M3 (fold expression, 3.0) (Cry3Bb1) and M2 (fold expression, 2.5) (Cry3) transformants of cotton. The confirmed transgenic plants were exposed to insect pests, pink bollworm (Pectinophora gossypiella), and army bollworm (Helicoverpa armigera). Bioassay results revealed that 60% mortality was observed against pink bollworm, and 75% mortality was observed against army bollworm in transgenic plants containing both Cry3Bb1 and Cry3 genes (M1 transgenic plants). In M2 transgenic plants containing only the Cry3Bb1 gene, the mortality was observed to be 40% in the pink bollworm population, whereas 45% mortality was observed in the army bollworm population. In the case of M3 transgenic plants containing single gene-Cry3, the mortality was 20% in the pink bollworm population, whereas 30% mortality was observed in the army bollworm population. Almost no mortality was observed in non-transgenic Eagle-2 control plants. Hence, the developed cotton transformants have improved resistance against chewing insect pests.

Potential Mechanisms of Transmission of Tick-Borne Viruses at the Virus-Tick Interface.

Ticks (Acari; Ixodidae) are the second most important vector for transmission of pathogens to humans, livestock, and wildlife. Ticks as vectors for viruses have been reported many times over the last 100 years. Tick-borne viruses (TBVs) belong to two orders (Bunyavirales and Mononegavirales) containing nine families (Bunyaviridae, Rhabdoviridae, Asfarviridae, Orthomyxovirida, Reoviridae, Flaviviridae, Phenuviridae, Nyamiviridae, and Nairoviridae). Among these TBVs, some are very pathogenic, causing huge mortality, and hence, deserve to be covered under the umbrella of one health. About 38 viral species are being transmitted by <10% of the tick species of the families Ixodidae and Argasidae. All TBVs are RNA viruses except for the African swine fever virus from the family Asfarviridae. Tick-borne viral diseases have also been classified as an emerging threat to public health and animals, especially in resource-poor communities of the developing world. Tick-host interaction plays an important role in the successful transmission of pathogens. The ticks' salivary glands are the main cellular machinery involved in the uptake, settlement, and multiplication of viruses, which are required for successful transmission into the final host. Furthermore, tick saliva also participates as an augmenting tool during the physiological process of transmission. Tick saliva is an important key element in the successful transmission of pathogens and contains different antimicrobial proteins, e.g., defensin, serine, proteases, and cement protein, which are key players in tick-virus interaction. While tick-virus interaction is a crucial factor in the propagation of tick-borne viral diseases, other factors (physiological, immunological, and gut flora) are also involved. Some immunological factors, e.g., toll-like receptors, scavenger receptors, Janus-kinase (JAK-STAT) pathway, and immunodeficiency (IMD) pathway are involved in tick-virus interaction by helping in virus assembly and acting to increase transmission. Ticks also harbor some endogenous viruses as internal microbial faunas, which also play a significant role in tick-virus interaction. Studies focusing on tick saliva and its role in pathogen transmission, tick feeding, and control of ticks using functional genomics all point toward solutions to this emerging threat. Information regarding tick-virus interaction is somewhat lacking; however, this information is necessary for a complete understanding of transmission TBVs and their persistence in nature. This review encompasses insight into the ecology and vectorial capacity of tick vectors, as well as our current understanding of the predisposing, enabling, precipitating, and reinforcing factors that influence TBV epidemics. The review explores the cellular, biochemical, and immunological tools which ensure and augment successful evading of the ticks' defense systems and transmission of the viruses to the final hosts at the virus-vector interface. The role of functional genomics, proteomics, and metabolomics in profiling tick-virus interaction is also discussed. This review is an initial attempt to comprehensively elaborate on the epidemiological determinants of TBVs with a focus on intra-vector physiological processes involved in the successful execution of the docking, uptake, settlement, replication, and transmission processes of arboviruses. This adds valuable data to the existing bank of knowledge for global stakeholders, policymakers, and the scientific community working to devise appropriate strategies to control ticks and TBVs.

Deep Neural Networks for Automatic Flower Species Localization and Recognition.

Deep neural networks are efficient methods of recognizing image patterns and have been largely implemented in computer vision applications. Object detection has many applications in computer vision, including face and vehicle detection, video surveillance, and plant leaf detection. An automatic flower identification system over categories is still challenging due to similarities among classes and intraclass variation, so the deep learning model requires more precisely labeled and high-quality data. In this proposed work, an optimized and generalized deep convolutional neural network using Faster-Recurrent Convolutional Neural Network (Faster-RCNN) and Single Short Detector (SSD) is used for detecting, localizing, and classifying flower objects. We prepared 2000 images for various pretrained models, including ResNet 50, ResNet 101, and Inception V2, as well as Mobile Net V2. In this study, 70% of the images were used for training, 25% for validation, and 5% for testing. The experiment demonstrates that the proposed Faster-RCNN model using the transfer learning approach gives an optimum mAP score of 83.3% with 300 and 91.3% with 100 proposals on ten flower classes. In addition, the proposed model could identify, locate, and classify flowers and provide essential details that include flower name, class classification, and multilabeling techniques.

Public opinion of the Irish "COVID Tracker" digital contact tracing App: A national survey.

Objective: This study aims to gather public opinion on the Irish "COVID Tracker" digital contact tracing (DCT) App, with particular focus on App usage, usability, usefulness, technological issues encountered, and potential changes to the App. Methods: A 35-item online questionnaire was deployed for 10 days in October 2020, 3 months after the launch of the Irish DCT App. Results: A total of 2889 completed responses were recorded, with 2553 (88%) respondents currently using the App. Although four in five users felt the App is easy to download, is easy to use and looks professional, 615 users (22%) felt it had slowed down their phone, and 757 (28%) felt it had a negative effect on battery life. Seventy-nine percent of respondents reported the App's main function is to aid contact tracing. Inclusion of national COVID-19 trends is a useful ancillary function according to 87% of respondents, and there was an appetite for more granular local data. Overall, 1265 (44%) respondents believed the App is helping the national effort, while 1089 (38%) were unsure. Conclusions: DCT Apps may potentially augment traditional contact tracing methods. Despite some reports of negative effects on phone performance, just 7% of users who have tried the App have deleted it. Ancillary functionality, such as up-to-date regional COVID-19, may encourage DCT App use. This study describes general positivity toward the Irish COVID Tracker App among users but also highlights the need for transparency on effectiveness of App-enabled contact tracing and for study of non-users to better establish barriers to use.

Genome-wide characterization and expression analysis of Erf gene family in cotton.

BACKGROUND: AP2/ERF transcription factors are important in a variety of biological activities, including plant growth, development, and responses to biotic and abiotic stressors. However, little study has been done on cotton's AP2/ERF genes, although cotton is an essential fibre crop. We were able to examine the tissue and expression patterns of AP2/ERF genes in cotton on a genome-wide basis because of the recently published whole genome sequence of cotton. Genome-wide analysis of ERF gene family within two diploid species (G. arboreum & G. raimondii) and two tetraploid species (G. barbadense, G. hirsutum) was performed. RESULTS: A total of 118, 120, 213, 220 genes containing the sequence of single AP2 domain were identified in G. arboreum, G. raimondii, G. barbadense and G. hirsutum respectively. The identified genes were unevenly distributed across 13/26 chromosomes of A and D genomes of cotton. Synteny and collinearity analysis revealed that segmental duplications may have played crucial roles in the expansion of the cotton ERF gene family, as well as tandem duplications played a minor role. Cis-acting elements of the promoter sites of Ghi-ERFs genes predict the involvement in multiple hormone responses and abiotic stresses. Transcriptome and qRT-PCR analysis revealed that Ghi-ERF-2D.6, Ghi-ERF-12D.13, Ghi-ERF-6D.1, Ghi-ERF-7A.6 and Ghi-ERF-11D.5 are candidate genes against salinity tolerance in upland cotton. CONCLUSION: Overwhelmingly, the present study paves the way to better understand the evolution of cotton ERF genes and lays a foundation for future investigation of ERF genes in improving salinity stress tolerance in cotton.

Digital Contact Tracing Apps for COVID-19: Development of a Citizen-Centered Evaluation Framework.

BACKGROUND: The silent transmission of COVID-19 has led to an exponential growth of fatal infections. With over 4 million deaths worldwide, the need to control and stem transmission has never been more critical. New COVID-19 vaccines offer hope. However, administration timelines, long-term protection, and effectiveness against potential variants are still unknown. In this context, contact tracing and digital contact tracing apps (CTAs) continue to offer a mechanism to help contain transmission, keep people safe, and help kickstart economies. However, CTAs must address a wide range of often conflicting concerns, which make their development/evolution complex. For example, the app must preserve citizens' privacy while gleaning their close contacts and as much epidemiological information as possible. OBJECTIVE: In this study, we derived a compare-and-contrast evaluative framework for CTAs that integrates and expands upon existing works in this domain, with a particular focus on citizen adoption; we call this framework the Citizen-Focused Compare-and-Contrast Evaluation Framework (C3EF) for CTAs. METHODS: The framework was derived using an iterative approach. First, we reviewed the literature on CTAs and mobile health app evaluations, from which we derived a preliminary set of attributes and organizing pillars. These attributes and the probing questions that we formulated were iteratively validated, augmented, and refined by applying the provisional framework against a selection of CTAs. Each framework pillar was then subjected to internal cross-team scrutiny, where domain experts cross-checked sufficiency, relevancy, specificity, and nonredundancy of the attributes, and their organization in pillars. The consolidated framework was further validated on the selected CTAs to create a finalized version of C3EF for CTAs, which we offer in this paper. RESULTS: The final framework presents seven pillars exploring issues related to CTA design, adoption, and use: (General) Characteristics, Usability, Data Protection, Effectiveness, Transparency, Technical Performance, and Citizen Autonomy. The pillars encompass attributes, subattributes, and a set of illustrative questions (with associated example answers) to support app design, evaluation, and evolution. An online version of the framework has been made available to developers, health authorities, and others interested in assessing CTAs. CONCLUSIONS: Our CTA framework provides a holistic compare-and-contrast tool that supports the work of decision-makers in the development and evolution of CTAs for citizens. This framework supports reflection on design decisions to better understand and optimize the design compromises in play when evolving current CTAs for increased public adoption. We intend this framework to serve as a foundation for other researchers to build on and extend as the technology matures and new CTAs become available.

Sentiment analysis of user feedback on the HSE's Covid-19 contact tracing app.

BACKGROUND: Digital Contact Tracing is seen as a key tool in reducing the propagation of Covid-19. But it requires high uptake and continued participation across the population to be effective. To achieve sufficient uptake/participation, health authorities should address, and thus be aware of, user concerns. AIM: This work manually analyzes user reviews of the Irish Heath Service Executive's (HSE) Contact Tracker app, to identify user concerns and to lay the foundations for subsequent, large-scale, automated analyses of reviews. While this might seem tightly scoped to the Irish context, the HSE app provides the basis for apps in many jurisdictions in the USA and Europe. METHODS: Manual analysis of (1287) user reviews from the Google/Apple playstores was performed, to identify the aspects of the app that users focused on, and the positive/negative sentiment expressed. RESULTS: The findings suggest a largely positive sentiment towards the app, and that users thought it handled data protection and transparency aspects well. But feedback suggests that users would appreciate more targeted feedback on the incidence of the virus, and facilities for more proactive engagement, like notifications that prompt users to submit their health status daily. Finally, the analysis suggests that the "android battery" issue and the backward-compatibility issue with iPhones seriously impacted retention/uptake of the app respectively. CONCLUSION: The HSE have responded to the public's desire for targeted feedback in newer versions, but should consider increasing the app's proactive engagement. The results suggest they should also raise the backward compatibility issue, regarding older iPhones, with Apple.

Transformation and Overexpression of Primary Cell Wall Synthesis-Related Zinc Finger Gene Gh_A07G1537 to Improve Fiber Length in Cotton.

Molecular interventions have helped to explore the genes involved in fiber length, fiber strength, and other quality parameters with improved characteristics, particularly in cotton. The current study is an extension and functional validation of previous findings that Gh_A07G1537 influences fiber length in cotton using a chromosomal segment substitution line MBI7747 through RNA-seq data. The recombinant Gh_A07G1537 derived from the MBI7747 line was over-expressed in CCRI24, a genotype with a low profile of fiber quality parameters. Putative transformants were selected on MS medium containing hygromycin (25mg/ml), acclimatized, and shifted to a greenhouse for further growth and proliferation. Transgene integration was validated through PCR and Southern Blot analysis. Stable integration of the transgene (ΔGh_A07G1537) was validated by tracking its expression in different generations (T0, T1, and T2) of transformed cotton plants. It was found to be 2.97-, 2.86-, and 2.92-folds higher expression in T0, T1, and T2 plants, respectively, of transgenic compared with non-transgenic cotton plants. Fiber quality parameters were also observed to be improved in the engineered cotton line. Genetic modifications of Gh_A07G1537 support the improvement in fiber quality parameters and should be appreciated for the textile industry.

Applications of copper and silver nanoparticles on wheat plants to induce drought tolerance and increase yield.

Drought adversely affects the yield, quality and nutritional value of the crop plants. Additionally, it causes unrest in society and economic loss to the farmers and governments. The present study involved the applications of nanoparticles to induce drought tolerance and improve yield in the wheat (Triticum aestivum L.) plants. Green chemical methods were used for the synthesis of silver nanoparticles (AgNPs) and copper nanoparticles (CuNPs). CuNPs were used in 0, 3, 5 and 7 mg/L and 0, 10, 20 and 30 mg/L of AgNPs were tested at -4, -6 and -8 bars osmotic potential in laboratory experiments and 40%, 60% and 80% field capacity (FC) in the glasshouse experiments were maintained. The solution culture experiments revealed significantly higher chlorophyll stability index (CSI), leaf succulence (LS) and leaf K (LK) content in plants treated with 03 mg/L of CuNPs and 10 mg/L of AgNPs, indicated the positive role of CuNPs and AgNPs in drought tolerance of wheat. A similar trend was observed for stomatal conductance (SC) and morphological parameters with the applications of Cu and Ag nanoparticles at different levels of field capacity. The results of this study provide the experimental evidence to use CuNPs and AgNPs to induce drought resistance and improve yield in the wheat plants by a satisfactory increase in nutrients uptake and water retention.

Pyramiding of cry toxins and methanol producing genes to increase insect resistance in cotton.

The idea of enhanced methanol production from cell wall by pectin methyl esterase enzymes (PME) combined with expression of cry genes from Bacillus thuringiensis as a strategy to improve insect pest control in cotton is presented. We constructed a cassette containing two cry genes (cry1Fa and Cry32Aa) and two pme genes, one from Arabidopsis thaliana (AtPME), and other from Aspergillus. niger (AnPME) in pCAMBIA1301 plant expression vector using CAMV-35S promoter. This construction was transformed in Eagle-2 cotton variety by using shoot apex-cut Agrobacterium-mediated transformation. Expression of cry genes and pme genes was confirmed by qPCR. Methanol production was measured in control and in the cry and pme transformed plants showing methanol production only in transformed plants, in contrast to the non-transgenic cotton plants. Finally, insect bioassays performed with transgenic plants expressing cry and pme genes showed 100% mortality for Helicoverpa armigera (cotton bollworm) larvae, 70% mortality for Pectinophora gossypiella (pink bollworm) larvae and 95% mortality of Earias fabia, (spotted bollworm) larvae, that was higher than the transgenic plants expressing only cry genes that showed 84%, 49% and 79% mortality, respectively. These results demonstrate that Bt. cry-genes coupled with pme genes are an effective strategy to improve the control of different insect pests.