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An SDS-NaOH-based method to isolate genome of recombinant adeno-associated virus vectors for physical titer measurement

Recombinant adeno-associated viruses (rAAVs) vectors are promising for their safety and sustained expression of genetic payloads across various tissues. These vectors consist of a protein capsid enclosing a 4.7 kb single-stranded DNA genome. Rapid and accurate determination of the physical titers of rAAV vector is crucial for quality control in rAAV manufacturing and precise drug dosage in clinical trials. To prepare vector DNA for genome titer assessment, it is essential to completely degrade unencapsulated DNA and dissociate the capsid. Conventional methods typically involve co-incubation with DNase I to degrade unencapsidated DNA, followed by co-incubation with Proteinase K to cleave protein shells. Here, we present a “Benzonase & SDS-NaOH" pretreatment as an effective alkaline lysis for releasing the vector DNA. In the presence of producer cell crude extract, Benzonase demonstrated superior efficacy in degrading unencapsidated DNA compared to DNase I. Additionally, the use of SDS-NaOH, effective at 65 °C for 30 min, significantly reduces the time required compared to that of Proteinase K at 56 °C for 2 hours. We also showed that the “Benzonase & SDS-NaOH" pretreatment is applicable for vector genome titration in rAAV production, harvest, and purified stock. Moreover, our method is effective for both scAAV and ssAAV forms and across all serotypes, including the thermally stable rAAV5. Overall, this method offers a rapid and straightforward solution to determine rAAV vector genome titers in both purified preparations and during the manufacturing process.

Network pharmacology and in silico analysis reveal Kochiae Fructus as a potential therapeutic against atopic dermatitis through immunomodulatory pathway interactions

Atopic dermatitis (AD), a chronic inflammatory disorder, poses significant therapeutic challenges owing to its complex pathophysiology, involving disrupted epidermal barrier function and immune dysregulation. This study investigated the therapeutic potential of Kochiae Fructus in AD treatment using bioinformatics, including network pharmacology and molecular docking techniques. We identified 19 key phytochemicals from Kochiae Fructus and 268 potential targets using the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) and SwissTarget Prediction. Using GeneCards, 1786 AD-related genes were retrieved, resulting in 116 intersecting gene targets for further analysis. Protein-protein interaction (PPI) networks and Molecular Complex Detection (MCODE) analyses highlighted 78 anti-AD key targets, including SRC, MAPK3, MAPK1, JUN, PIK3CA, ESR1, PTGS2, PTPN11, IL-6, and ALOX5, among the top ten anti-AD core targets. Gene ontology (GO) enrichment analysis revealed that Kochiae Fructus affects biological processes and molecular functions, such as positive regulation of the apoptotic response, inflammatory response, and hormone-mediated signaling pathways, which may be associated with its anti-AD effects. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the C-type lectin receptor signaling pathway is the main pathway involved in the anti-AD effects of Kochiae Fructus, which interacts with a notably larger number of anti-AD core targets and plays a direct role in intensifying crucial inflammatory and immune responses in the heart of AD pathogenesis. Molecular docking demonstrated robust binding affinities of key phytochemicals, particularly ecdysterone and 11,14-eicosadienoic acid, to the anti-AD core targets. Molecular dynamics simulations of over 1000 ns confirmed the stability and potential efficacy of these interactions. Hence, this study underscores the therapeutic potential of Kochiae Fructus in AD management, offering a mechanistic basis for its clinical application and paving the way for novel anti-AD strategies that leverage TCM phytochemicals.

Journal targeted by paper mill still grappling with the aftermath years later

Research on plasma arc flame length detection technology based on region of interest

With the rapid advancement of metal 3D printing technology, there is a growing demand for spherical metal powder as a primary material for 3D printing. The process technology that ensures the production of high-quality spherical metal powder has become a focal area of research for numerous enterprises and research institutions globally. In the conventional plasma rotating electrode method for powder production, the feed speed of the servo feeding mechanism is manually predetermined, leading to potential variations in the distance between the end face of the metal rod and the plasma gun that generates the plasma arc. Such inconsistency can compromise the quality of the metal powder produced and pose safety hazards if the gap between the metal rod and the plasma gun is too narrow. To address these issues, this study presents a novel plasma arc length detection system based on the concept of the region of interest. The proposed system leverages image processing technology for efficiently detecting the plasma arc length. By incorporating image detection within the region of interest alongside an arc length correction function, the system enhances real-time performance and detection precision. Additionally, real-time monitoring of the detection site is enabled through KingView. Experimental findings indicate that the image target area post plasma arc detection exhibits well-defined edges, clear brightness, and minimal noise, thereby meeting the prerequisites for subsequent image processing and monitoring tasks. The corrected plasma arc length averages around 40mm, with a detection error of less than 1mm when compared to the desired controlled plasma arc length. Moreover, the length variation remains relatively stable, thus fulfilling the measurement criteria. Over time, the detected plasma arc length exhibits negligible fluctuations, suggesting consistent proximity between the plasma gun and the end face of the metal rod during the melting process. The controller can dynamically control the feed speed of the servo feeding mechanism according to the detected plasma arc length, ensuring a constant distance between the plasma arc and the end face of the metal rod throughout the powder production process, thus aligning with practical industrial requirements.

The early origins of bone-tool manufacturing traditions by hominins 1.5 million years ago

An examination of the interrelationships among NASA-TLX dimensions utilizing the DEMATEL method

The NASA TLX is a survey-based method widely used to evaluate cognitive workload across six specific dimensions related to an employee’s tasks. While academic research recognizes that these dimensions may have unequal contributions to mental workload, they are often weighted using multi-criteria decision-making techniques. However, prior studies have not investigated possible relationships between the dimensions. The main objective of this paper is to explore the interconnections and dependencies among these dimensions. This study distinguishes itself by employing the Decision-Making Trial and Evaluation Laboratory (DEMATEL) technique to clarify these relationships and dependencies, and to examine how interactions between dimensions shift under different threshold conditions. The research includes three distinct impact diagrams tailored for individuals performing tasks with varying levels of cognitive workload. By considering the interdependencies among the NASA TLX dimensions, this study offers a significant advancement in the field, as these interrelationships could greatly influence the derived weights. This theoretical contribution has the potential to be groundbreaking in the realm of survey-based mental workload assessment techniques, such as the NASA TLX.

Rethinking Shadowing for Aspiring Physicians

Repeatability and genetic advances in early maturing maize hybrid trials conducted under Striga-infested and non-infested conditions

In sub-Saharan Africa (SSA), maize (Zea mays L.) is both a cash crop and an important staple crop. However, Striga hermonthica infection constrains its production and productivity. A total of 159 hybrids from 21 international trials were evaluated under Striga-infested (STRINF) and Striga non-infested (STRNON) conditions at Mokwa and Abuja, Nigeria, from 2010 to 2021. The data were used to (i) determine the genetic enhancements in grain yield and Striga adaptive traits and (ii) assess the repeatability of the trials in identification of promising hybrids. Significant annual genetic gains in grain yield of 3.40% and 3.71% with increases of 76.87 and 127.02 kg ha − 1 yr − 1 were recorded under STRINF and STRNON conditions, respectively. The genetic gains in grain yield were associated with 3.04 and 2.25% decreases in Striga damage ratings at 8 and 10 weeks after planting (WAP), respectively, and 1.27% in the number of emerged Striga plants at 10 WAP. The results indicated that ears per plant and flowering dates had the highest consistency in repeatability estimates while the number of emerged Striga plants at 8 and 10 WAP recorded the lowest consistency in repeatability estimates. Generally, substantial progress has been achieved with a good level of repeatability estimates for the early maturing maize hybrid trials evaluated under STRINF and STRNON conditions.Those results have demonstrated that the breeding strategies adopted during the 12-year breeding period have been effective, and that the potential of the trials in the exploration of the genetic potential of the hybrids for commercialization in the SSA for food security and poverty alleviation.

Ménétrier’s Disease

How does the brain control consciousness? This deep-brain structure

Author Correction: B-cell-specific checkpoint molecules that regulate anti-tumour immunity

The balance evaluation systems test (BESTest), mini-BESTest and brief-BESTest as clinical tools to assess balance control across different populations: A reliability generalization meta-analysis

Background The Balance Evaluation Systems Test (BESTest) and two abbreviated versions, Mini-BESTest and Brief-BESTest are used to assess functioning of balance control systems. Its reliability across different populations remains to be determined. Objective The present study followed reliability generalization procedures to estimate an average internal consistency and inter and intra-rater reliability for the BESTest, Mini-BESTest and Brief-BEStest. In this study, the heterogeneity of reliability coefficients in each instrument is evaluated. If heterogeneity is significant, a moderator analysis is performed to identify the characteristic which explains such variability. Methods A search of the PubMed, Embase, PsycINFO, Web of Science, Scopus and CINAHL databases was carried out to February 10th 2024. Two reviewers independently selected empirical studies published in English or Spanish that applied the BESTest, Mini-BESTest and/or Brief-BESTest and reported any reliability coefficient and/or internal consistency with data at hand. Results Sixty-four studies reported any reliability estimate BESTest, Mini-BESTest and/or Brief-BESTest scores (N. = 5225 participants). Mean Cronbach alpha for the Mini-BESTest and Brief-BESTest (total score = 0.92) indicating no variability in estimated internal consistency. Likewise, no variability was obtained for inter-rater and intra-rater mean agreement of the BESTest (ICC = 0.97; 0.94), Mini-BESTest (ICC = 0.95; 0.94) and Brief-BESTest (ICC = 0.96; 0.95). Mean scores, standard deviation of scores, mean age, gender, population type, mean history of the disorder, disease, raters´ experience, number of raters, rater formation, continent of study and design type presented statistically significant relationships with ICC and/or Cronbach´s alpha for BESTest and the two abbreviated versions. Conclusions The mean intraclass correlations and Cronbach alpha obtained for BESTest, Mini-BESTest and Brief-BESTest exhibited an excellent inter and intra-rater reliability and internal consistency. The average reliability obtained three scales adequate to be applied for screening balance problems in different populations. Some continuous and categorical moderator variables increase reliability and internal consistency of these scales.

Medical School Grading — Is “Good Enough” Good Enough?

Take Nature’s poll: How will Trump’s policies affect US science?

Effects of the Family Nurse Partnership on all eligible mothers: a data linkage cohort study in England

Background An intensive programme of home visiting, the Family Nurse Partnership (FNP), is received by around one in four first-time adolescent mothers in selected areas in England. During home visits, nurses support mothers to make choices about healthy pregnancies, improving child development, and fulfilling their own aspirations and ambitions. Evidence is needed of the wider effects of the FNP, including for mothers not enrolled in the programme (who might experience unintended effects). We evaluated child and maternal outcomes for all eligible mothers giving birth before, during, and after the period in which FNP was active in local areas. Methods We created a linked cohort of 237,185 eligible mothers, aged 13-19, who gave birth between April 2010 and March 2019, and who had a first antenatal booking appointment (or a date of 28 completed weeks of gestation, if missing) when FNP was active in their area. We used administrative hospital data to identify unplanned maternal/child hospitalisations up to 2 years after birth for children born and mothers delivering before, during and after FNP was active. Generalised linear models were used to adjust for background regional time trends, maternal characteristics, and clustering of outcomes within residential areas. Results We found no evidence of differences in unplanned hospital admissions for children born during the FNP period (36.9% versus 36.0%, relative risk [RR] 1.01; 95% CI 0.99-1.02), or after FNP was active (37.1%, RR 1.0; 95% CI 0.95-1.06), compared with those born before FNP was active. There was no evidence of differences in child admissions for maltreatment/injury-related diagnoses or for maternal admissions for adversity-related diagnoses. Conclusion Child and maternal outcomes were similar before, during and after FNP active periods, suggesting that the FNP did not have a wider impact on outcomes in all eligible mothers, including those not participating in the FNP.

Left Atrial Appendage Closure after Ablation for Atrial Fibrillation

Potential of saccharomyces cerevisiae fermentation-derived postbiotic technology in mitigating multiple drug-resistant Salmonella enterica serovars in an in vitro broiler cecal model

Diamond V Original XPC® is a Saccharomyces cerevisiae fermentation-derived postbiotic technology (SCFP) designed to interact synergistically with the animal to provide health benefits by enhancing immune function, supporting digestive integrity and absorption, and maintaining gastrointestinal (GIT) microbial balance in the host. The current study investigated the effects of 1.25% SCFP on multidrug-resistant (MDR) Salmonella serovars: S. Typhimurium (ATCC 14028), S. Enteritidis, S. Infantis, S. Heidelberg, S. Typhimurium DT104, and S. Reading, and shifts in cecal microbiota populations. Using an anaerobic in vitro poultry cecal model, cecal contents were inoculated with ~ 108 colony forming units (CFU) of MDR Salmonella serovars and incubated for 24 h at 37°C anaerobically. The treatments included: control group consisting of 0.2 g of crushed poultry feed, and a treatment group 0.25 g of feed +  1.25% inclusion of Original XPC® (SCFP). The SCFP significantly reduced five of the six serovars: S. Typhimurium ATCC, S. Enteritidis, S. Infantis, S. Heidelberg, and S. Reading (P <  0.05). Time significantly impacted S. Typhimurium DT104 reduction (P <  0.001). The most significant decrease was observed for S. Enteritidis (3.9 log10 CFU/mL), followed by S. Heidelberg (3.8 log10 CFU/mL), S. Infantis (3.4 log10 CFU/mL), S. Typhimurium ATCC (3 log10 CFU/mL), and S. Reading (1.8 log10 CFU/mL) compared to controls that averaged approximately 1 log10 CFU/mL reduction. Microbiota analysis at 24 h involved genomic DNA extraction, amplification using custom dual-indexed primers, and sequencing on the Illumina MiSeq platform. Sequencing data were analyzed using QIIME2-2021.11. S. Infantis and S. Heidelberg inoculated samples were the only groups that significantly enhanced microbial richness and evenness with SCFP addition at 24 h (P <  0.05). Pairwise comparisons revealed that samples inoculated with S. Reading and S. Typhimurium DT104 exhibited a minor change in microbial composition with SCFP, compared to other serovars that demonstrated increased microbial diversity with SCFP. Additionally, S. Infantis and S. Heidelberg inoculated samples exhibited phylogenetic diversity and microbial abundance with SCFP compared to controls at 24 h (P <  0.05). Lachnospiraceae CHKCI001 was significantly more abundant in SCFP-treated samples compared to controls (ANCOM, P <  0.05), suggesting SCFP impact on cecal fermenters and production of fermentation end products that may impact the ecosystem and inhibit pathogen growth. Although various serovars may exhibit somewhat different responses, SCFP effectively mitigated multiple MDR serovars of Salmonella under in vitro incubation conditions.

NEJM at AHA — Left Atrial Appendage Closure after Ablation for Atrial Fibrillation

Exclusive: Trump White House directs NIH to study ‘regret’ after transgender people transition

Strengths, weaknesses, opportunities and threats of mobile applications in undergraduate nursing: a scoping review protocol

Aim To describe the protocol for a scoping review on the main strengths, weaknesses, opportunities and threats of mobile applications in undergraduate nursing. Background Some studies have shown that mobile applications have been objects of increasing interest in nursing education. Although these technologies offer potential benefits to the educational process, their implementation and use are not always without challenges and implications. In this sense, the SWOT analysis will be used to summarize the strengths, weaknesses, opportunities and threats of those technologies. Design Scoping review protocol, conducted in accordance with JBI guidelines. Method This is a protocol for a scoping review to be carried out following the JBI guidelines, in three phases and in the following data sources: Nursing Database, Latin American and Caribbean Literature in Health Sciences, Medical Literature Analysis and Retrieval System Online, Scopus, Web of Science and ScienceDirect, later, Google Scholar and, finally, in the references of the primarily collected studies. The inclusion criteria are studies that addressed the topic of mobile applications in Nursing teaching, available in full, free of charge and without time or language cut-off. The exclusion criteria are studies that did not address the topic, opinion articles, letters to the editor and editorials. The results will be presented through tables and a SWOT chart, with the study variables. Expected Results The results will cover the characterization of the sample studies, followed by a SWOT chart containing the main strengths, weaknesses, opportunities and threats of mobile applications in undergraduate nursing. Conclusion The SWOT analysis will identify strengths and weaknesses, promote self-knowledge and strategies for mobile applications in undergraduate nursing. It can also assist in decision-making, communication with partners and publication, improving scientific research.