Efficiency, Specificity and Temperature Sensitivity of Cas9 and Cas12a RNPs for DNA-free Genome Editing in Plants

Delivery of genome editing reagents using CRISPR-Cas ribonucleoproteins (RNPs) transfection offers several advantages over plasmid DNA-based delivery methods, including reduced off-target editing effects, mitigation of random integration of non-native DNA fragments, independence of vector constructions, and less regulatory restrictions. Compared to the use in animal systems, RNP-mediated genome editing is still at the early development stage in plants. In this study, we established an efficient and simplified protoplast-based genome editing platform for CRISPR-Cas RNP delivery, and then evaluated the efficiency, specificity, and temperature sensitivity of six Cas9 and Cas12a proteins. Our results demonstrated that Cas9 and Cas12a RNP delivery resulted in genome editing frequencies (8.7–41.2%) at various temperature conditions, 22°C, 26°C, and 37°C, with no significant temperature sensitivity. LbCas12a often exhibited the highest activities, while AsCas12a demonstrated higher sequence specificity. The high activities of CRISPR-Cas RNPs at 22° and 26°C, the temperature preferred by plant transformation and tissue culture, led to high mutagenesis efficiencies (34.0–85.2%) in the protoplast-regenerated calli and plants with the heritable mutants recovered in the next generation. This RNP delivery approach was further extended to pennycress (Thlaspi arvense), soybean (Glycine max) and Setaria viridis with up to 70.2% mutagenesis frequency. Together, this study sheds light on the choice of RNP reagents to achieve efficient transgene-free genome editing in plants.

Equipping Property Graduates for the Digital Age

Bridging the gaps between property graduates’ attributes and industry expectations has been touted to be the key driver of sustainable development for the next generation of the workforce. This study investigated property-related knowledge and skills from the perspective of property employers and graduates in Australia in the digital age by (1) identifying the most and least important knowledge and skillsets in the property sector; (2) examining the strategies used to develop the required knowledge and skillsets; and (3) ascertaining if there are differences in the knowledge and skillsets perceived to be necessary between employers and graduates. A questionnaire survey was undertaken across property employers affiliated with the Royal Institution of Chartered Surveyors (RICS) and graduates of an Australian university. The results showed that problem solving and time management are the most important knowledge and skills for property employers and graduates, respectively. Notably, there were statistically significant differences in the perceived importance of course directors maintaining close communication with employers, real-life case study-based school assessment, the use of various course delivery methods, inviting guest speakers, and internship training while studying between employers and graduates. This finding implies that Australian property professionals are yet to fully embrace technology in the digital age.

What Are We Measuring When We Evaluate Digital Interventions for Improving Lifestyle? A Scoping Meta-Review

Background: Lifestyle Medicine (LM) aims to address six main behavioral domains: diet/nutrition, substance use (SU), physical activity (PA), social relationships, stress management, and sleep. Digital Health Interventions (DHIs) have been used to improve these domains. However, there is no consensus on how to measure lifestyle and its intermediate outcomes aside from measuring each behavior separately. We aimed to describe (1) the most frequent lifestyle domains addressed by DHIs, (2) the most frequent outcomes used to measure lifestyle changes, and (3) the most frequent DHI delivery methods.Methods: We followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA-ScR) Extension for Scoping Reviews. A literature search was conducted using MEDLINE, Cochrane Library, EMBASE, and Web of Science for publications since 2010. We included systematic reviews and meta-analyses of clinical trials using DHI to promote health, behavioral, or lifestyle change.Results: Overall, 954 records were identified, and 72 systematic reviews were included. Of those, 35 conducted meta-analyses, 58 addressed diet/nutrition, and 60 focused on PA. Only one systematic review evaluated all six lifestyle domains simultaneously; 1 systematic review evaluated five lifestyle domains; 5 systematic reviews evaluated 4 lifestyle domains; 14 systematic reviews evaluated 3 lifestyle domains; and the remaining 52 systematic reviews evaluated only one or two domains. The most frequently evaluated domains were diet/nutrition and PA. The most frequent DHI delivery methods were smartphone apps and websites.Discussion: The concept of lifestyle is still unclear and fragmented, making it hard to evaluate the complex interconnections of unhealthy behaviors, and their impact on health. Clarifying this concept, refining its operationalization, and defining the reporting guidelines should be considered as the current research priorities. DHIs have the potential to improve lifestyle at primary, secondary, and tertiary levels of prevention—but most of them are targeting clinical populations. Although important advances have been made to evaluate DHIs, some of their characteristics, such as the rate at which they become obsolete, will require innovative research designs to evaluate long-term outcomes in health.

Hepatitis E, Schistosomiasis and Echinococcosis–Prevalence in a Cohort of Pregnant Migrants in Germany and Their Influence on Fetal Growth Restriction

Background: Infections, as well as adverse birth outcomes, may be more frequent in migrant women. Schistosomiasis, echinococcosis, and hepatitis E virus (HEV) seropositivity are associated with the adverse pregnancy outcomes of fetal growth restriction and premature delivery. Methods: A cohort study of 82 pregnant women with a history of migration and corresponding delivery of newborns in Germany was conducted. Results: Overall, 9% of sera tested positive for anti-HEV IgG. None of the patients tested positive for anti-HEV IgM, schistosomiasis, or echinococcus serology. Birth weights were below the 10th percentile for gestational age in 8.5% of the neonates. No association between HEV serology and fetal growth restriction (FGR) frequency was found. Conclusions: In comparison to German baseline data, no increased risk for HEV exposure or serological signs of exposure against schistosomiasis or echinococcosis could be observed in pregnant migrants. An influence of the anti-HEV serology status on fetal growth restriction could not be found.

Role of Nanoparticles in Cancer Therapy

The rapid growth of nanotechnology towards the development of nanomedicines has improved cancer treatment. Nanomedicine provides the opportunity to implement complex and targeted multifunctional strategies. Today, nanoparticles (NPs) have many uses in a number of scientific fields. In recent years, it has been repeatedly reported that NPs hold a significant place in the regulation of modern medicine by implementing a varying number of clinical approaches like drug carrying substances, genetic material delivery to tumors, as well as in radiography as a contrast media agent. Various nanomaterials based on organic, inorganic, lipid or glycan compounds, and synthetic polymers have been used to develop and improve new cancer treatments. In this chapter, the authors discussed the role of NPs in cancer treatment among various anticancer drug delivery methods.

Clinical Skills Development in the Virtual Learning Environment

The rapid transition to distance learning in response to the unexpected SARS-CoV-2/COVID-19 pandemic led to disruption of clinical skills development, which are typically conducted face-to-face. Consequently, faculty adapted their courses, using a multitude of active learning modalities, to meet student learning objectives in the didactic and experiential settings. Strategies and considerations to implement innovative delivery methods and address potential challenges are elucidated. Furthermore, integration of a layered learning approach may allow for more broad perspectives and allow additional interactions and feedback, which is especially necessary in the virtual environment.

Course Model Redesign for Continuity of Instruction

The purpose of the chapter is to illustrate instructional models that were implemented by Samford University McWhorter School of Pharmacy to comply with COVID-19 social distancing restrictions. While the second half of Spring 2020 was completely online (statewide shutdown), the university remained open in a hybrid manner for the 2020-21 academic year. There are three sections in the chapter: didactic, interprofessional, and advanced pharmacy practice experiences. The didactic section discusses course delivery methods and active learning, office hours, remote testing, student feedback, and contingency planning. The interprofessional section illustrates some of the school's synchronous and asynchronous interprofessional learning activities before and during the COVID-pandemic, as well as interprofessional education assessment methods. The last section of the chapter discusses how advanced pharmacy practice experience “direct patient care” was redefined, examples of the experiences, and contingency plans that were put into place to ensure on-time graduation for the classes of 2020 and 2021 pharmacy students.

Media and Human Resource Management

Easily mentioned, but not that much categorically identified, is the infrastructure beyond huge communication networks providers. Undoubtedly, the commitment for people of a certain country, in their transformation from citizens of a definite jurisdiction to netizens of a global community seeking advanced training and instruction with interactive educational TV, heavily relies on top management support over complex issues, starting from governmental initiatives and ardent information technology support along vigorous settlements with vendors, private funds, and international conglomerates. What the student body seeks, in national scale, is how to enhance the delivery methods of its learning, from face-to-face methodologies to self-paced learning, and recently, even further to live e-learning. For the first time, however, in-depth skill building, support in a range of practical subjects, animated knowledge acquisition, peer networking, and action-planning have been introduced as managerial educative aspects, along information and entertainment, in the world of broadcasting.