Genome editing: An insight into disease resistance, production efficiency, and biomedical applications in livestock.

One of the primary concerns for the survival of the human species is the growing demand for food brought on by an increasing global population. New developments in genome-editing technology present promising opportunities for the growth of wholesome and prolific farm animals. Genome editing in large animals is used for a variety of purposes, including biotechnology to improve food production, animal health, and pest management, as well as the development of animal models for fundamental research and biomedicine. Genome editing entails modifying genetic material by removing, adding, or manipulating particular DNA sequences from a particular locus in a way that does not happen naturally. The three primary genome editors are CRISPR/Cas 9, TALENs, and ZFNs. Each of these enzymes is capable of precisely severing nuclear DNA at a predetermined location. One of the most effective inventions is base editing, which enables single base conversions without the requirement for a DNA double-strand break (DSB). As reliable methods for precise genome editing in studies involving animals, cytosine and adenine base editing are now well-established. Effective zygote editing with both cytosine and adenine base editors (ABE) has resulted in the production of animal models. Both base editors produced comparable outcomes for the precise editing of point mutations in somatic cells, advancing the field of gene therapy. This review focused on the principles, methods, recent developments, outstanding applications, the advantages and disadvantages of ZFNs, TALENs, and CRISPR/Cas9 base editors, and prime editing in diverse lab and farm animals. Additionally, we address the methodologies that can be used for gene regulation, base editing, and epigenetic alterations, as well as the significance of genome editing in animal models to better reflect real disease. We also look at methods designed to increase the effectiveness and precision of gene editing tools. Genome editing in large animals is used for a variety of purposes, including biotechnology to improve food production, animal health, and pest management, as well as the development of animal models for fundamental research and biomedicine. This review is an overview of the existing knowledge of the principles, methods, recent developments, outstanding applications, the advantages and disadvantages of zinc finger nucleases (ZFNs), transcription-activator-like endonucleases (TALENs), and clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR/Cas 9), base editors and prime editing in diverse lab and farm animals, which will offer better and healthier products for the entire human race.

Mammalian IRE1α dynamically and functionally coalesces with stress granules.

Upon endoplasmic reticulum (ER) stress, activation of the ER-resident transmembrane protein kinase/endoribonuclease inositol-requiring enzyme 1 (IRE1) initiates a key branch of the unfolded protein response (UPR) through unconventional splicing generation of the transcription factor X-box-binding protein 1 (XBP1s). Activated IRE1 can form large clusters/foci, whose exact dynamic architectures and functional properties remain largely elusive. Here we report that, in mammalian cells, formation of IRE1α clusters is an ER membrane-bound phase separation event that is coupled to the assembly of stress granules (SGs). In response to different stressors, IRE1α clusters are dynamically tethered to SGs at the ER. The cytosolic linker portion of IRE1α possesses intrinsically disordered regions and is essential for its condensation with SGs. Furthermore, disruption of SG assembly abolishes IRE1α clustering and compromises XBP1 mRNA splicing, and such IRE1α-SG coalescence engenders enrichment of the biochemical components of the pro-survival IRE1α-XBP1 pathway during ER stress. Our findings unravel a phase transition mechanism for the spatiotemporal assembly of IRE1α-SG condensates to establish a more efficient IRE1α machinery, thus enabling higher stress-handling capacity.

Multiple RNA Profiling Reveal Epigenetic Toxicity Effects of Oxidative Stress by Graphene Oxide Silver Nanoparticles in-vitro.

Introduction: The increasing industrial and biomedical utilization of graphene oxide silver nanoparticles (GO-AgNPs) raises the concern of nanosafety: exposure to the AgNPs or GO-AgNPs increases the generation of reactive oxygen species (ROS), causes DNA damage and alters the expression of whole transcriptome including mRNA, miRNA, tRNA, lncRNA, circRNA and others. Although the roles of different RNAs in epigenetic toxicity are being studied during the last decade, but still we have little knowledge about the role of circle RNAs (circRNAs) in epigenetic toxicity. Methods: Rabbit fetal fibroblast cells (RFFCs) were treated with 0, 8, 16, 24, 32 and 48 µg/mL GO-AgNPs to test the cell viability and 24 µg/mL GO-AgNPs was selected as the experimental dose. After 24 h treatment with 24 µg/mL GO-AgNPs, the level of ROS, malondialdehyde (MDA), superoxide dismutase (SOD), intracellular ATP, glutathione peroxidase (GPx), and glutathione reductase (Gr) were measured in the RFFCs. High-throughput whole transcriptome sequencing was performed to compare the expression of circRNAs, long non-coding RNAs (lncRNA) and mRNA between 24 µg/mL GO-AgNPs-treated RFFCs and control cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to validate the accuracy of circRNA sequencing data. Bioinformatics analyses were performed to reveal the potential functional roles and related pathways of differentially expressed circRNAs, lncRNA and mRNA and to construct a circRNA-miRNA-mRNA interaction network. Results: We found that 57 circRNAs, 75 lncRNAs, and 444 mRNAs were upregulated while 35 circRNAs, 21 lncRNAs, and 186 mRNAs were downregulated. These differentially expressed genes are mainly involved in the transcriptional mis-regulation of cancer through several pathways: MAPK signaling pathway (circRNAs), non-homologous end-joining (lncRNAs), as well as PPAR and TGF-beta signaling pathways (mRNAs). Conclusion: These data revealed the potential roles of circRNAs in the GO-AgNPs induced toxicity through oxidative damage, which would be the basis for further research to determine their roles in the regulation of different biological processes.

Identification of circular RNAs expression pattern in caprine fetal fibroblast cells exposed to a chronic non-cytotoxic dose of graphene oxide-silver nanoparticle nanocomposites.

The widespread use of graphene oxide-silver nanoparticle nanocomposites (GO-AgNPs) in biomedical sciences is increasing the chances of human and animal exposure to its chronic non-toxic doses. Exposure to AgNPs-related nanomaterials may result in the negative effect on the dam, fetus and offspring. However, there are only little available information for profound understanding of the epigenetic alteration in the cells and animals caused by low-dose chronic exposure of GO-AgNPs. The present study investigated the effect of 0.5 µg/mL GO-AgNPs for 10 weeks on the differential expression of circular RNAs (circRNAs) in caprine fetal fibroblast cells (CFFCs), and this dose of GO-AgNPs did not affect cell viability and ROS level. We predicted the functions of those differentially expressed (DE) circRNAs in CFFCs by bioinformatics analysis. Furthermore, we validated the expression of ten DE circRNAs using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) to ensure the reliability of the sequencing data. Our results showed that the DE circRNAs may potentially regulate the GO-AgNPs-inducing epigenetic toxicity through a regulatory network consisted of circRNAs, miRNAs and messenger RNAs (mRNAs). Therefore, the epigenetics toxicity is essential to assess the biosafety level of GO-AgNPs.

Adipocyte IRE1α promotes PGC1α mRNA decay and restrains adaptive thermogenesis.

Adipose tissue undergoes thermogenic remodeling in response to thermal stress and metabolic cues, playing a crucial role in regulating energy expenditure and metabolic homeostasis. Endoplasmic reticulum (ER) stress is associated with adipose dysfunction in obesity and metabolic disease. It remains unclear, however, if ER stress-signaling in adipocytes mechanistically mediates dysregulation of thermogenic fat. Here we show that inositol-requiring enzyme 1α (IRE1α), a key ER stress sensor and signal transducer, acts in both white and beige adipocytes to impede beige fat activation. Ablation of adipocyte IRE1α promotes browning/beiging of subcutaneous white adipose tissue following cold exposure or ß3-adrenergic stimulation. Loss of IRE1α alleviates diet-induced obesity and augments the anti-obesity effect of pharmacologic ß3-adrenergic stimulation. Notably, IRE1α suppresses stimulated lipolysis and degrades Ppargc1a messenger RNA through its RNase activity to downregulate the thermogenic gene program. Hence, blocking IRE1α bears therapeutic potential in unlocking adipocytes' thermogenic capacity to combat obesity and metabolic disorders.

Predicting Avoidable Emergency Department Visits Using the NHAMCS Dataset.

Despite the important role avoidable emergency department (ED) visits play in healthcare costs and quality of care, there has been little work in development of predictive models to identify patients likely to present with an avoidable ED visit. We use a conservative definition of 'avoidable' ED visits defined as visits that do not require diagnostic or screening services, procedures, or medications, and were discharged home to classify visits as avoidable. Models trained using data from emergency departments across the US yielded a training AUC of 0.723 and a testing AUC of 0.703. Models trained using the full dataset were tested on demographic groups (race, gender, insurance status), finding comparable performance between white/black patients and male/female with reductions in performance in Hispanic populations and patients with Medicaid. Predictors strongly associated with non-avoidable ED visits included increased age, increasing number of total chronic diseases, and general as well as digestive symptoms. Reasons for visit stated as injuries and psychiatric symptoms influenced the model to predict an avoidable visit.

The ER stress sensor inositol-requiring enzyme 1α in Kupffer cells promotes hepatic ischemia-reperfusion injury.

Hepatic ischemia/reperfusion (I/R) injury is an inflammation-mediated process arising from ischemia/reperfusion-elicited stress in multiple cell types, causing liver damage during surgical procedures and often resulting in liver failure. Endoplasmic reticulum (ER) stress triggers the activation of the unfolded protein response (UPR) and is implicated in tissue injuries, including hepatic I/R injury. However, the cellular mechanism that links the UPR signaling to local inflammatory responses during hepatic I/R injury remains largely obscure. Here, we report that IRE1α, a critical ER-resident transmembrane signal transducer of the UPR, plays an important role in promoting Kupffer-cell-mediated liver inflammation and hepatic I/R injury. Utilizing a mouse model in which IRE1α is specifically ablated in myeloid cells, we found that abrogation of IRE1α markedly attenuated necrosis and cell death in the liver, accompanied by reduced neutrophil infiltration and liver inflammation following hepatic I/R injury. Mechanistic investigations in mice as well as in primary Kupffer cells revealed that loss of IRE1α in Kupffer cells not only blunted the activation of the NLRP3 inflammasome and IL-1ß production, but also suppressed the expression of the inducible nitric oxide synthase (iNos) and proinflammatory cytokines. Moreover, pharmacological inhibition of IRE1α's RNase activity was able to attenuate inflammasome activation and iNos expression in Kupffer cells, leading to alleviation of hepatic I/R injury. Collectively, these results demonstrate that Kupffer cell IRE1α mediates local inflammatory damage during hepatic I/R injury. Our findings suggest that IRE1α RNase activity may serve as a promising target for therapeutic treatment of ischemia/reperfusion-associated liver inflammation and dysfunction.

Evaluation of Patient Experience During Virtual and In-Person Urgent Care Visits: Time and Cost Analysis.

Urgent care (UC) is one of the fastest growing venues of health care delivery for nonemergent conditions. This study compared the patient experience during virtual and in-person urgent care visits. We conducted a cross-sectional study of patients with the same diagnosis during Virtual Urgent Care (VUC) and in-person UC visits with the same diagnosis with regard to time and cost over a period of one year. We recorded and analyzed 16 685 urgent care visits: In-person UC (n = 14 734), VUC (n = 1262). Significant differences were found in the average total time for a visit in an in-person UC (70.89 minutes), and VUC (9.38 minutes). The average total cost of VUC ($49) and in-person UC ($142.657) differed significantly. Significant difference was found between UC turnaround time and VC turnaround time (Dependent variable (DV): 53.77, P < .01). We found significant differences in cost and time between in favor of virtual visits. Our findings suggest additional policy reform to expand the use of virtual care among target populations to improve access, reduce costs, meet the needs of patients, and reduce emergency department visits.

How does medical scribes' work inform development of speech-based clinical documentation technologies? A systematic review.

OBJECTIVE: Use of medical scribes reduces clinician burnout by sharing the burden of clinical documentation. However, medical scribes are cost-prohibitive for most settings, prompting a growing interest in developing ambient, speech-based technologies capable of automatically generating clinical documentation based on patient-provider conversation. Through a systematic review, we aimed to develop a thorough understanding of the work performed by medical scribes in order to inform the design of such technologies. MATERIALS AND METHODS: Relevant articles retrieved by searching in multiple literature databases. We conducted the screening process following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) in guidelines, and then analyzed the data using qualitative methods to identify recurring themes. RESULTS: The literature search returned 854 results, 65 of which met the inclusion criteria. We found that there is significant variation in scribe expectations and responsibilities across healthcare organizations; scribes also frequently adapt their work based on the provider's style and preferences. Further, scribes' job extends far beyond capturing conversation in the exam room; they also actively interact with patients and the care team and integrate data from other sources such as prior charts and lab test results. DISCUSSION: The results of this study provide several implications for designing technologies that can generate clinical documentation based on naturalistic conversations taking place in the exam room. First, a one-size-fits-all solution will be unlikely to work because of the significant variation in scribe work. Second, technology designers need to be aware of the limited role that their solution can fulfill. Third, to produce comprehensive clinical documentation, such technologies will likely have to incorporate information beyond the exam room conversation. Finally, issues of patient consent and privacy have yet to be adequately addressed, which could become paramount barriers to implementing such technologies in realistic clinical settings. CONCLUSIONS: Medical scribes perform complex and delicate work. Further research is needed to better understand their roles in a clinical setting in order to inform the development of speech-based clinical documentation technologies.

Understanding the Association Between Electronic Health Record Satisfaction and the Well-Being of Nurses: Survey Study.

BACKGROUND: Intensive care unit (ICU) nurses experience high levels of burnout related to the high-stress environment. Management of electronic health records (EHR) is a contributing factor to physician burnout. However, limited research has established the relationship between the nurse's well-being and EHR use. OBJECTIVE: The objective of this study was to examine the association between EHR use and the well-being of nurses. METHODS: We surveyed registered nurses employed at a major Southeastern medical center in the United States about their demographics, experience with EHRs, satisfaction with EHRs, and elements of well-being. The correlation between subgroup demographics and survey questions was examined using Kendall and Fisher tests. RESULTS: A total of 113 ICU registered nurses responded to the survey, of which 93 (82.3%) were females. The population had a mean age of 35.18 years (SD 10.65). A significant association was found between satisfaction and well-being scores, where higher EHR satisfaction was associated with higher self-reported well-being (correlation 0.35, P<.001). Nurses who were unhappy with the time spent in EHR use compared with direct patient care reported higher levels of stress (P<.001) and isolation (P=.009). Older nurses reported higher dissatisfaction with the amount of time spent on EHR tasks related to direct patient care compared to younger nurses (P<.001). CONCLUSIONS: Although nurses reported acceptable satisfaction scores with EHR use, deeper analysis suggests that EHR indirectly affects the well-being of nurses. These findings strongly indicate that lower EHR satisfaction can impact the well-being of nurses. More research is needed to optimize the nurse-EHR experience through more user-centered design approaches.

Unusually Dispersed AgI Quantum Dots For Efficient HTL-Free CH3NH3PbI3 Photovoltaics.

The utilization of quantum dots (QDs) to improve the performance of perovskite solar cells is attracting much attention due to their unique optical and electronic properties. Most of QDs have to be prepared in advance and then incorporated into the perovskite hosts, which could not ensure the maintenance of their QD characteristics. In this work, we intelligently developed an in situ preparation strategy to disperse AgI QDs homogeneously in the perovskite host for the MAPbI3:AgI(QDs) cross-blended layer directly on indium tin oxide (ITO) via a common and convenient spin-coating process. We combine transmission electron microscopy, X-ray photoelectron spectroscopy, and Raman techniques to demonstrate the cross-blended MAPbI3:AgI(QDs) structure in the final perovskite devices. Furthermore, a series of simply inverted ITO/MAPbI3:AgI(QDs)/PCBM/Ag devices have been designed and fabricated. The photovoltaic performance of these solar cells shows significantly improved short-circuit current density (Jsc) and a champion power conversion efficiency of 16.41% even without a hole transport layer. The current technique induced the crystal growth toward high-quality perovskite films with a homogeneous structure, good crystallinity, less grain boundaries and defects, increased optical path length, and uniform thickness for better solar cell performance. Besides, the impact of the current strategy also lies in an accommodation effect of the hole collection at the ITO side induced by AgI QDs, which modifies the Fermi level of perovskite films, leading to significantly decreased level difference in the Fermi level/work function between the perovskite layer and ITO substrates by ultraviolet photoelectron spectra analysis. More importantly, the charge carrier dynamics of such novel MAPbI3:AgI(QDs) structures were also scrutinized by transient photovoltage analysis.

Analyzing the Demographics of Virtual Care Users.

Using telemedicine to provide care is an attractive alternative for patients. However, few studies have examined the demographics of the patients using virtual care. In this paper, we investigate the demographic features of the Virtual Care (VC) users; Age, gender, roles, and preferred methods of communication are studied.

Advancing health equity and access using telemedicine: a geospatial assessment.

INTRODUCTION: Health disparity affects both urban and rural residents, with evidence showing that rural residents have significantly lower health status than urban residents. Health equity is the commitment to reducing disparities in health and in its determinants, including social determinants. OBJECTIVE: This article evaluates the reach and context of a virtual urgent care (VUC) program on health equity and accessibility with a focus on the rural underserved population. MATERIALS AND METHODS: We studied a total of 5343 patient activation records and 2195 unique encounters collected from a VUC during the first 4 quarters of operation. Zip codes served as the analysis unit and geospatial analysis and informatics quantified the results. RESULTS: The reach and context were assessed using a mean accumulated score based on 11 health equity and accessibility determinants calculated for each zip code. Results were compared among VUC users, North Carolina (NC), rural NC, and urban NC averages. CONCLUSIONS: The study concluded that patients facing inequities from rural areas were enabled better healthcare access by utilizing the VUC. Through geospatial analysis, recommendations are outlined to help improve healthcare access to rural underserved populations.

Factors Determining Patients' Choice Between Mobile Health and Telemedicine: Predictive Analytics Assessment.

BACKGROUND: The solution to the growing problem of rural residents lacking health care access may be found in the use of telemedicine and mobile health (mHealth). Using mHealth or telemedicine allows patients from rural or remote areas to have better access to health care. OBJECTIVE: The objective of this study was to understand factors influencing the choice of communication medium for receiving care, through the analysis of mHealth versus telemedicine encounters with a virtual urgent clinic. METHODS: We conducted a postdeployment evaluation of a new virtual health care service, Virtual Urgent Clinic, which uses mHealth and telemedicine modalities to provide patient care. We used a multinomial logistic model to test the significance and predictive power of a set of features in determining patients' preferred method of telecare encounters-a nominal outcome variable of two levels (mHealth and telemedicine). RESULTS: Postdeployment, 1403 encounters were recorded, of which 1228 (87.53%) were completed with mHealth and 175 (12.47%) were telemedicine encounters. Patients' sex (P=.004) and setting (P<.001) were the most predictive determinants of their preferred method of telecare delivery, with significantly small P values of less than .01. Pearson chi-square test returned a strong indication of dependency between chief concern and encounter mediums, with an extremely small P<.001. Of the 169 mHealth patients who responded to the survey, 154 (91.1%) were satisfied by their encounter, compared with 31 of 35 (89%) telemedicine patients. CONCLUSIONS: We studied factors influencing patients' choice of communication medium, either mHealth or telemedicine, for a virtual care clinic. Sex and geographic location, as well as their chief concern, were strong predictors of patients' choice of communication medium for their urgent care needs. This study suggests providing the option of mHealth or telemedicine to patients, and suggesting which medium would be a better fit for the patient based on their characteristics.

Multivariate Analysis of Physicians' Practicing Behaviors in an Urgent Care Telemedicine Intervention.

When assessing the characteristics and performance of telemedicine interventions, most studies followed a patient- centric approach, leaving the telemedicine providers' role out of consideration. As a result, little was known about the demographics and prescription pattern of telemedicine physicians, the knowledge of which is integral to a holistic evaluation of the virtual delivery of accountable care. To fill this gap, our study explored how physicians' traits and encounter-specific characteristics correlate with prescription outcomes, using multivariate analyses. Significant inter-physician variation in prescription behaviors was observed and analyzed in sub-groups. The average Virtual Urgent Care physician's prescription likelihood was 69% with a mean prescription count of 0.98; male physicians and primary care providers tended to prescribe both more often and with a greater number of medications. This study called attention to the quality and reproducibility of telemedicine providers' prescription decision and warned the likely absence of well-defined practice guidelines for delivering virtual care.