OBJECTIVE: It has been shown that variable compared to fixed task prioritization during dual task practice more effectively improves motor (i.e., postural control) and cognitive (i.e., memory) performance in older adults. However, it is unclear whether this finding is also valid in young adults. Thus, the present study examined the effect of fixed (allocate equal priority on both tasks) versus variable (vary priority between both tasks) priority during short-term motor-cognitive dual task practice on single and dual task performance in healthy young adults (age range: 20-30 years). RESULTS: During two days of practice, significant improvements of motor (i.e., balance task: reduced root mean square error; p < 001, ηp2 = .72) and cognitive (i.e., arithmetic task: increased serial three subtractions; p < .001, ηp2 = .78) task performance were observed and that was irrespective of group ("fixed priority" and "variable priority"). Further, the statistical analysis of post-practice single and dual task performance revealed no significant differences between groups, irrespective of task (i.e., motor or cognitive). This indicates that in young as opposed to old adults, single and dual task performance improvements are independent of task prioritization (i.e., fixed or variable priority) during short-term motor-cognitive dual task practice.

OBJECTIVE: The mosquito transmitted dengue virus (DENV) the causative agent of dengue fever (DF) remains a significant public health burden in many countries. Thailand, along with many countries in Asia and elsewhere, has a long history of using traditional medicines to combat febrile diseases such as DF. Screening bioactive compounds from traditional medicines reported to have antipyretic or anti-inflammatory activity may lead to the development of potent antivirals. In this study oroxylin A (OA), a flavonoid derivative found in Oroxylum indicum (commonly called the Indian trumpet flower or tree of Damocles), was screened for antiviral activity towards DENV. RESULTS: Cytotoxicity analysis in BHK-21 cells showed a 50% cytotoxic concentration (CC50) of 534.17 µM. The compound showed no direct virucidal activity towards DENV, and pre-treatment of cells had no effect on virus production. A deficit was seen in virus production when cells were post-infection treated with oroxylin A. Under conditions of post-infection treatment, the EC50 value was 201.1 µM, giving a selectivity index (SI) value of 2.66. Accumulation of DENV E protein inside the cell was seen under conditions of post-infection treatment, suggesting that oroxylin A may exert some effects at the virus assembly/egress stages of the replication cycle.

OBJECTIVE: The determination of the location of quantitative trait loci (QTL) (i.e., QTL mapping) is essential for identifying new genes. Various statistical methods are being incorporated into different QTL mapping functions. However, statistical errors and limitations may often occur in a QTL mapping, implying the risk of false positive errors and/or failing to detect a true positive QTL effect. We simulated the power to detect four simulated QTL in tomato using cim() and stepwiseqtl(), widely adopted QTL mapping functions, and QTL.gCIMapping(), a derivative of the composite interval mapping method. While there is general agreement that those three functions identified simulated QTL, missing or false positive QTL were observed, which were prevalent when more realistic data (such as smaller population size) were provided. RESULTS: To address this issue, we developed postQTL, a QTL mapping R workflow that incorporates (i) both cim() and stepwiseqtl(), (ii) widely used R packages developed for model selection, and (iii) automation to increase the accuracy, efficiency, and accessibility of QTL mapping. QTL mapping experiments on tomato F2 populations in which QTL effects were simulated or calculated showed advantages of postQTL in QTL detection.

OBJECTIVES: To investigate free-living, accelerometry-derived step cadence and walking strategy parameters in 263 adult women (19-56 years) within a rural African setting. Participants were categorised into weight groups: Under-to-Normal Weight (UW/NW: < 25 kg/m2), Overweight-to-Obese (OW/OB: ≥ 25 kg/m2). From the minute-by-minute uni-axial accelerometry data, outcomes describing physical activity intensity, step volume, step cadence and step bouts were extracted. In addition, walking pattern parameters for step bout length and step cadence were determined. RESULTS: Average step volume was 13,568 steps/day, and > 85% of participants were classified as active-to-highly-active. Overall, ≈ 45% of daily steps was accumulated in the low-to-moderate intensity range. Peak cadence indices were higher in the UW/NW group (p ≤ 0.0112). For both groups, 75% of steps were accumulated in bouts > 15 min, and 95% of bouts were accumulated at 1-39 steps/min. The UW/NW group employed a more varied step cadence, and higher cadences contributed more to step accumulation than the OW/OB group (p ≤ 0.05). There were no significant group differences in bout length strategy parameters (p ≥ 0.0861). Despite no difference between the weight groups in step volume, there were differences in some step cadence indices which reflect higher step intensities, and in cadence strategies chosen to accumulate steps.

OBJECTIVE: The present study aimed to investigate the effects of nano-curcumin supplementation on adipokines levels and clinical signs in obese and overweight patients with migraine. RESULTS: Forty-four patients with episodic migraine participated in this clinical trial and were divided into two groups nano-curcumin (80 mg/day) and the control group over 2-month period. At the baseline and the end of the research, the serum levels of MCP-1, Resistin, and Visfatin were measured using the ELISA method. In addition, the headache attack frequencies, severity, and duration of pain were recorded. The results of the present study showed that nano-curcumin can significantly reduce MCP-1 serum levels in the nano-curcumin supplemented group (P = 0.015, size effect = 13.4%). In the case of resistin and visfatin, nano-curcumin supplementation exerted no statistically significant changes in serum levels (P > 0.05). Nano-curcumin also significantly reduced the attack frequencies, severity, and duration of headaches (P < 0.05). These findings indicate that targeting curcumin can be a promising approach to migraine management. However, further comprehensive human trials are needed to confirm these findings. TRIAL REGISTRATION: This study was registered in the Iranian Registry of Clinical Trials (IRCT) with ID number: IRCT20160626028637N2 on the date 2020-07-10.

OBJECTIVE: To introduce salvage prostatectomy in Denmark. Prior to this, no national curative treatment for recurrent prostate cancer following radiation therapy existed in Denmark. This pilot study represent our initial experiences and the feasibility of performing salvage robot-assisted radical prostatectomy for true local, high-risk recurrence after initial therapy with external beam radiation for high-risk prostate cancer. RESULTS: Five patients underwent sRARP between April 2020 and July 2021. All patients were discharged within 48 h and no major complications were observed within 3 months. All patients had unmeasurable PSA (< 0.1 ng/ml) at follow-up 6 months after surgery. One patient with longer follow-up than 6 months experienced biochemical recurrence. At 3-months follow-up all patients reported considerable incontinence, at 6-month follow-up, pad usage decreased to 1 or 2 pads daily. Based on our initial results, the idea to introduce sRARP as a nationwide option remains and further patients will be included to establish the true role of sRARP in patients with recurrence after primary radiotherapy for PCa.

OBJECTIVE: Toxoplasma gondii is a ubiquitous parasite of medical and veterinary importance; however, there exists no cure for chronic toxoplasmosis. Metabolic enzymes required for the production and maintenance of tissue cysts represent promising targets for novel therapies. Here, we use reverse genetics to investigate the role of Toxoplasma phosphoglucomutase 1, PGM1, in Toxoplasma growth and cystogenesis. RESULTS: We found that disruption of pgm1 did not significantly affect Toxoplasma intracellular growth and the lytic cycle. pgm1-defective parasites could differentiate into bradyzoites and produced cysts containing amylopectin in vitro. However, cysts produced in the absence of pgm1 were significantly smaller than wildtype. Together, our findings suggest that PGM1 is dispensable for in vitro growth but contributes to optimal Toxoplasma cyst development in vitro, thereby necessitating further investigation into the function of this enzyme in Toxoplasma persistence in its host.

OBJECTIVE: Person-to-person transmission can occur during outbreaks of verotoxigenic Escherichia coli (VTEC), however the impact of this transmission route is not well understood. This study aimed to examine the role of person-to-person transmission during a VTEC outbreak, and how targeting this route may reduce outbreak size. A deterministic compartmental model describing a VTEC outbreak was constructed and fit to data from a 2008 outbreak in Ontario, Canada. Using the best-fit model, simulations were run to calculate the: reduction in transmission rate after implementing interventions, proportion of cases infected through both transmission routes, and number of cases prevented by interventions. Latin hypercube sensitivity analysis was conducted to examine the sensitivity of the outbreak size to the model parameters. RESULTS: Based on the best-fit model, ~ 14.25% of the cases likely arose due to person-to-person transmission. Interventions reduced this transmission rate by ~ 73%, causing a reduction in outbreak size of ~ 17% (47 cases). Sensitivity analysis showed that the model was highly sensitive to changes in all parameters of the model. The model demonstrates that person-to-person could be an important transmission route during VTEC outbreaks. Targeting this route of transmission through hand hygiene and work exclusions could reduce the final outbreak size.

OBJECTIVES: We assessed Case Fatality Rate (CFR) of COVID-19 as an indicator to situate the performance of Nigeria relative to other selected countries. We obtained case fatality rates of different countries from data sets available from open-sources. The CFRs were calculated as the rate of deaths compared with total cases. The values were compared with Nigeria's COVID-19 CFR. Other relevant statistical comparisons were also conducted. RESULTS: The worst performing countries with regards to CFR in descending order were Yemen (19.5%), Peru (9.0%) Mexico (7.6%), Sudan (7.4%) and Ecuador (6.3%) while the best performing nations were Bhutan (0.11%), Burundi (0.19%), Iceland (0.20%), Laos (0.21%) and Qatar (0.25%). The CFR of Nigeria was 1.39% which falls below the 50th percentile. Other comparison done showed significant difference in the CFR values between countries similar to Nigeria and countries that are dissimilar when HDI is used. (Mann-Whitney U test 126.0, p = 0.01). The trend of the CFR in Nigeria showed a steady decline and flattening of the CFR curve which does not seem to be affected by the spikes in the daily declared cases.

One major source of exhaustion for researchers is the redundant paperwork of three different documents-research papers, ethics review applications, and research grant applications-for the same research plan. This is a wasteful and redundant process for researchers, and it has a more direct impact on the career development of early-career researchers. Here, we propose a trinity review system based on Registered Reports that integrates scientific, ethics, and research funding reviews. In our proposed trinity review system, scientific and ethics reviews are undertaken concurrently for a research protocol before running the study. After the protocol is approved in principle through these review processes, a funding review will take place, and the researchers will begin their research. Following the experiments or surveys, the scientific review will be conducted on a completed version of the paper again, including the results and discussions (i.e., the full paper), and the full paper will be published once it has passed the second review. This paper provides the brief process of the trinity review system and discusses the need for and benefits of the proposed system. Although the trinity review system only applies to a few appropriate disciplines, it helps improve reproducibility and integrity.

OBJECTIVE: The fungus Metarhizium brunneum produces ergot alkaloids of the lysergic acid amide class, most abundantly lysergic acid α-hydroxyethylamide (LAH). Genes for making ergot alkaloids are clustered in the genomes of producers. Gene clusters of LAH-producing fungi contain an α/β hydrolase fold protein-encoding gene named easP whose presence correlates with LAH production but whose contribution to LAH synthesis in unknown. We tested whether EasP contributes to LAH accumulation through gene knockout studies. RESULTS: We knocked out easP in M. brunneum via a CRISPR/Cas9-based approach, and accumulation of LAH was reduced to less than half the amount observed in the wild type. Because LAH accumulation was reduced and not eliminated, we identified and mutated the only close homolog of easP in the M. brunneum genome, a gene we named estA. An easP/estA double mutant did not differ from the easP mutant in lysergic acid amide accumulation, indicating estA had no role in the pathway. We conclude EasP contributes to LAH accumulation but is not absolutely required. Either a gene encoding redundant function and lacking sequence identity with easP resides outside the ergot alkaloid synthesis gene cluster, or EasP plays an accessory role in the synthesis of LAH.

OBJECTIVE: To reanalyze a clinical trial on the effectiveness of a Brief Intervention (BI) delivered by non-professionals to reduce risky alcohol drinking. Our previous null-hypothesis test of the effects of the BI yielded a 'non-significant' p-value, yet remained uninformative. Here we use the Bayesian paradigm which allows for expressing the probability of different effect sizes to better inform public policy decisions. RESULTS: The posterior probability of the odds of risky drinking at follow-up favored a marked effect of the BI, with 96% of the probability mass being less than OR = 1, and 84% being less than OR = 0.8. Our findings show that there is a high probability that the BI delivered by health technicians lowered risky alcohol use. The posterior distributions of the BI's effects are presented to help contextualize the evidence for policy making in Chile.

OBJECTIVE: Social scientists have suggested two typical ways of acquiring social power: dominance approach (gaining social power by applying violence, coercion, threat, and punishment) and prestige approach (gaining admiration and liking by demonstrating competence and sharing experience and knowledge). However, little is known about how people recognize and evaluate the differentiated process of the approaches, and even less about the early development of these processes. In the current study, 5-6-year old children heard stories about pairs comprising a dominance-based and a prestige-based powerholder, chose one of the powerholders as their friend and leader, and predicted which powerholder will gain the contested resources. RESULTS: Compared to a dominance-based powerholder, children were more likely to choose a prestige-based powerholder as a friend and leader in different situations. Moreover, children predicted that prestige-based powerholders, and not dominance-based powerholders, would gain contested resources. These findings suggest that since childhood, human beings tend to be biased to not only judge prestige-based aspects as socially preferable, but also endorse the prestige-based powerholders' priority to possess valuable resources, which subsequently strengthens their high social status. These early childhood preferences can be instrumental in providing more harmonious environments for children in educational and daily contexts.

Research integrity, an essential precept of scientific inquiry and discovery, comprises norms such as Rigor, Reproducibility, and Responsibility (the 3R's). Over the past decades, numerous issues have arisen that challenge the reliability of scientific studies, including irreproducibility crises, lack of good scientific principles, and erroneous communications, which have impacted the public's trust in science and its findings. Here, we highlight one important component of research integrity that is often overlooked in the discussion of proposals for improving research quality and promoting robust research; one that spans from the lab bench to the dissemination of scientific work: responsible science communication. We briefly outline the role of education and institutions of higher education in teaching the tenets of good scientific practice and within that, the importance of adequate communications training. In that context, we present our framework of responsible science communication that we live by and teach to our students in courses and workshops that are part of the Johns Hopkins Bloomberg School of Public Health R3 Center for Innovation in Science Education.

OBJECTIVE: The relationship of sagittal spinal shape in the pathogenesis of adolescent idiopathic scoliosis (AIS) is recognised. What is not clear is the relationship between the sagittal shape of those without scoliosis and the potential development of AIS, including the greater prevalence in females. The use of a new parameter, the kyphosis-lordosis (KL) difference, was developed to explore this further. RESULTS: The KL difference was calculated for 117 males and 79 females over seven years with 831 measures made. For females, the KL difference, between the ages of 9 and 12 ½ years, decreases from 5° to nearly 0° until starting to climb again from the age of 14 years, back to 5° by the age of 16 ½ years. For males, there is a gradual decline from 9° at age 9 years to 5° at age 17 years. Both age and sex were statistically significant in the development of the parameter. When comparing to previously published data around the true, de-rotated, sagittal shape of the scoliotic spine, the KL difference has utility in explaining the female predominance in the prevalence of AIS. This adds to the weight of evidence behind understanding why AIS develops.

OBJECTIVES: In children, achieving an acceptable degree of accuracy from dietary or physical activity (PA) assessments remains a challenge. Children tend to overestimate their time spent in daily PA and underestimate their dietary intake of fat and sugar. Because parents play a key role in family lifestyle decisions, including children's food choices and PA levels, it is important to investigate the responses of parents regarding their children's lifestyle habits. We aimed to develop a Self-Administered Parental Assessment of Children's Lifestyle (SAPLACL) questionnaire and test its feasibility and reliability in 191 parents (29 fathers and 162 mothers). RESULTS: For each part of the questionnaire, the rate of missing or improper responses ranged from 0 to 24%. The highest proportion of problems in understanding was reported for the dietary intake dimension, especially for snacking in front of the TV. Some difficulty was also found regarding the question on leisure PA. Test-retest agreement was observed in 54.7-100% of the respondents. Overall, the kappa coefficients were favorable. Thus, the parent self-report questionnaire is a valid and accurate tool for analyzing children's lifestyle habits in France.

OBJECTIVES: Mitral regurgitation (MR) and microvascular obstruction (MVO) are common complications of myocardial infarction (MI). This study aimed to investigate the association between MR in ST-elevation MI (STEMI) subjects with MVO post-reperfusion. STEMI subjects undergoing primary percutaneous intervention were enrolled. Cardiovascular magnetic resonance (CMR) imaging was performed within 48-hours of initial presentation. 4D flow images of CMR were analysed using a retrospective valve tracking technique to quantify MR volume, and late gadolinium enhancement images of CMR to assess MVO. RESULTS: Among 69 patients in the study cohort, 41 had MVO (59%). Patients with MVO had lower left ventricular (LV) ejection fraction (EF) (42 ± 10% vs. 52 ± 8%, P < 0.01), higher end-systolic volume (98 ± 49 ml vs. 73 ± 28 ml, P < 0.001) and larger scar volume (26 ± 19% vs. 11 ± 9%, P < 0.001). Extent of MVO was associated with the degree of MR quantified by 4D flow (R = 0.54, P = 0.0003). In uni-variate regression analysis, investigating the association of CMR variables to the degree of acute MR, only the extent of MVO was associated (coefficient = 0.27, P = 0.001). The area under the curve for the presence of MVO was 0.66 (P = 0.01) for MR > 2.5 ml. We conclude that in patients with reperfused STEMI, the degree of acute MR is associated with the degree of MVO.

OBJECTIVE: Exercise has been demonstrated to be beneficial for improving physical capacity and quality of life in people with scleroderma, although knowledge of its impact on the respiratory system is limited. This study evaluated the impact of therapist-oriented home rehabilitation (TOHR) on impulse oscillometry (IOS) and lung ultrasound (LUS) findings in patients with scleroderma-associated interstitial lung disease (ILD). RESULTS: Twelve women with scleroderma underwent spirometry, IOS, and LUS before and after performing TOHR. Regarding spirometry, a normal pattern and restrictive damage were observed in five (41.7%) and seven (58.3%) participants pre-TOHR and post-TOHR, respectively. For IOS, an abnormal result was detected in nine (75%) pre-TOHR participants and six (50%) post-TOHR participants. Heterogeneity of resistance between 4-20 Hz (R4-R20) > 20% of the predicted value was observed in eight (66.7%) pre-TOHR participants and three (25%) post-TOHR participants (P = 0.031). An abnormal LUS result was observed in nine (75%) participants both pre-TOHR and post-TOHR. The main change observed was B-lines > 2, which was noted in nine (75%) participants both pre-TOHR and post-TOHR. Our findings suggest that TOHR for women with scleroderma-associated ILD improves the resistance and reactance measured by IOS, including small airway disease. Trial Registration ClinicalTrials.gov ID: NCT05041868 Registered on: 13th September 2021.

Poor quality medical research causes serious harms by misleading healthcare professionals and policymakers, decreasing trust in science and medicine, and wasting public funds. Here we outline underlying problems including insufficient transparency, dysfunctional incentives, and reporting biases. We make the following recommendations to address these problems: Journals and funders should ensure authors fulfil their obligation to share detailed study protocols, analytical code, and (as far as possible) research data. Funders and journals should incentivise uptake of registered reports and establish funding pathways which integrate evaluation of funding proposals with initial peer review of registered reports. A mandatory national register of interests for all those who are involved in medical research in the UK should be established, with an expectation that individuals maintain the accuracy of their declarations and regularly update them. Funders and institutions should stop using metrics such as citations and journal's impact factor to assess research and researchers and instead evaluate based on quality, reproducibility, and societal value. Employers and non-academic training programmes for health professionals (clinicians hired for patient care, not to do research) should not select based on number of research publications. Promotions based on publication should be restricted to those hired to do research.

OBJECTIVE: Portal mesenchymal cells induce the epithelial differentiation of the bile ducts in the developing liver via one of the Delta-Notch signaling components, JAGGED1. Although this differential induction is crucial for normal liver physiology as its genetic disorder (Alagille syndrome) causes jaundice, the molecular mechanism behind JAGGED1 expression remains unknown. Here, we searched for upstream regulatory transcription factors of JAGGED1 using an integrated bioinformatics method. RESULTS: According to the DoRothEA database, which integrates multiple lines of evidence on the relationship between transcription factors and their downstream target genes, three transcription factors were predicted to be upstream of JAGGED1: SLUG, SOX2, and EGR1. Among these, SLUG and EGR1 were enriched in ACTA2-expressing portal mesenchymal cells in two previously reported human fetal liver single-cell RNA-seq datasets. JAGGED1-expressing portal mesenchymal cells tended to express SLUG rather than EGR1, supporting that SLUG induced JAGGED1 expression. Together with the higher confidentiality of SLUG (DoRothEA level A) over EGR1 (DoRothEA level D), we concluded that SLUG was one of the most important candidate transcription factors upstream of JAGGED1. These results add mechanistic insights into the developmental biology of how portal mesenchymal cells support biliary development in the liver.

OBJECTIVE: We aimed to evaluate cytocompatibility of hyaluronic acid (HA) and gelatin (Gela) conjugation with phenolic groups (Phs) via enzyme-mediated crosslinking. Phenolic moieties were substituted on the backbone of HA (HA-Ph) and Gela (Gela-Ph) and subsequently were subjected for horseradish peroxidase crosslinking in the presence of H2O2 as an electron donor to create a stable hybrid microenvironment for cellular behavior and cartilage tissue engineering. RESULTS: Successful synthesis of biopolymers confirmed by NRM and UV-Vis spectrophotometry. The physical characteristic of hydrogels including mechanical properties and water contact angle of hydrogels enhanced with addition of Gela-Ph in HA-based hydrogel. The Gela-Ph showed longest gelation time and highest degradation rate. The cellular studies showed cells did not attach to HA-Ph hydrogel. While, proper cell attachment and proliferation observed on blend hydrogel surface compared with the neat hydrogels which interpret by the existence of cell-adhesive motifs of utilized Gela-Ph in this hydrogel. The encapsulated cells in HA-Ph hydrogel were spheroid and just maintained their viability. Hydrogels containing Gela-Ph, the cells were spindle shape with high degrees of cytoplasmic extension. Overall, the results suggest that hybrid biomimetic hydrogel can provide a superior biological microenvironment for chondrocytes in 3D cartilage tissue engineering.

OBJECTIVE: The admixture of domestic pig into wild boar populations is controlled until now, by cytogenetic analysis. Even if a first-generation hybrid animal is discernable because of its 37-chromosome karyotype, the cytogenetic method is not applicable in the case of advanced intercrosses. The aim of this study is therefore to evaluate the use of SNP (Single Nucleotide Polymorphism) markers as an alternative technology to characterize recent or past hybridization between the two sub-species. The final goal would be to develop a molecular diagnostic tool. DATA DESCRIPTION: The Geneseek Genomic Profiler High-Density porcine beadchip (GGP70KHD, Illumina, USA), comprising 68,516 porcine SNPs, was used on a set of 362 wild boars with diverse chromosomal statuses collected from different areas and breeding environments in France. We generated approximately 62,192-64,046 genotypes per wild boar. The present dataset might be useful for the community (i) for developing molecular tools to evaluate the admixture of domestic pig into wild boar populations, and (ii) for genetic diversity studies including wild boar species or phylogeny analyses of Suidae populations. Raw data files and a processed matrix data file were deposited in the ArrayExpress at European Molecular Biology Laboratory-European Bioinformatics Institute (EMBL-EBI) data portal under accession number E-MTAB-10591.

OBJECTIVE: Bactofilins can assemble into polymeric structures and play important roles in cell shape maintenance, chromosome segregation and motility. Bacillus subtilis bactofilins BacE and BacF were shown to be important for swimming motility in strain PY79, and single gene deletions were reported to be lethal, in contrast to a double bacEF deletion. RESULTS: Extending this work, we show that motility defects vary between different B. subtilis strains, with strain 168 showing no defect in motility, and 3610 showing delayed induction of swimming. Generation of single gene deletions in PY79 was possible by transferring corresponding deletions from 168. In the natural isolate 3610, gene deletions also showed a negative effect on biofilm formation, revealing an additional function for BacE and BacF. A spontaneous arising suppressor mutation in PY79 was mapped to the flhO gene, a constituent of the flagellum, which obtained an 18 amino acid extension at its C-terminus. Our findings show that bactofilin gene deletions lead to different motility phenotypes dependent on the strain background, and affect biofilm formation in the natural isolate 3610. Our data reinforce the idea of a connection between bactofilins and motion via the flagellum, and suggest that they operate in a switch like manner.

Erratum in BMC Res Notes. 2022 Jul 25;15(1):262.

OBJECTIVE: The detection and monitoring of DNA methylation status in circulating tumor cell DNA (ctDNA) provides critical insights into cancer diagnosis and progression. The methylation status of the Dickkopf-related protein 3 (DKK3) promoter region is correlated with the metastasis and recurrence of multiple cancers. Thus, detecting the methylation status via non-invasive methods is essential for the diagnosis and prognosis of cancers. Using a droplet digital polymerase chain reaction approach, we have developed a highly sensitive and quantitative measurement of methylated and unmethylated DKK3 derived from circulating cell-free DNA (ccfDNA). RESULTS: We confirmed the specificity of droplet digital methylation specific polymerase chain reaction (ddMSP). We selected the optimal bisulfite conversion method using commercially available kits. We validated the ddMSP analysis system by analyzing the methylation status of genomic DNA extracted from cultured mesothelioma cells and mesothelial cells. Our system quantified approximately 30 copies of cell-free DNA per 4 mL, which is sufficient for detecting ctDNA. Finally, we quantified methylated and unmethylated DKK3 copies in ccfDNA from 21 patients with malignant mesothelioma.

OBJECTIVE: The aim is to describe and reflect upon potentially pandemic-related impact on self-assessments of active ageing. As part of the baseline data collection in the Prospective RELOC-AGE (ClinicalTrials.gov NCT04765696) study, telephone interviews, including the University of Jyvaskyla Active Aging Scale (UJACAS) were conducted with 820 people 55 years or older listed with an interest of relocation at three housing companies in Sweden. Field notes alongside the interviews focused on two topics: (1) how respondents reasoned and replied to the questions included in the UJACAS; (2) whether there were specific items that seemed to be affected by the pandemic. RESULTS: For four items (Participating in events, Exercising, Maintaining friendships, Getting to know new people), recurrent comments indicated that respondents had been affected by the pandemic situation regarding one or more of the facets in UJACAS: will to act, ability to act, opportunity to act, or frequency or extent of doing the activity. Opportunities to act was most frequently commented on as a factor affected by restricted participation in activities. As Prospective RELOC-AGE is a longitudinal study focused on associations between housing, relocation and active ageing, it is imperative to consider the potential pandemic-related impact on baseline data in future analyses.

OBJECTIVE: Patterns of cryopreservation of explanted skull bone flaps have long been a matter of debate, in particular the appropriate temperature of storage. To the best of our knowledge no study to date has compared the microbiological profile and the infection potential of skull bone flaps cryostored at the same institution at disparate degrees for neurosurgical purposes. In the context of our clinical trial DRKS00023283, we performed a bacterial culture of explanted skull bone flaps, which were cryopreserved lege artis at a temperature of either - 23 °C or - 80 °C after a decompressive hemicraniectomy. In a further step, we contaminated the bone fragments in a s uspension with specific pathogens (S. aureus, S. epidermidis and C. acnes, Colony forming unit CFU 103/ml) over 24 h and conducted a second culture. RESULTS: A total of 17 cryopreserved skull flaps (8: - 23 °C; 9: - 80 °C) explanted during decompressive hemicraniectomies performed between 2019 and 2020 as well as 2 computer-aided-designed skulls (1 vancomycin-soaked) were analyzed. Median duration of cryopreservation was 10.5 months (2-17 months). No microorganisms were detected at the normal bacterial culture. After active contamination of our skull flaps, all samples showed similar bacterial growth of above-mentioned pathogens; thus, our study did not reveal an influence of the storage temperature upon infectious dynamic of the skulls.

OBJECTIVE: Environmental enrichment is used to treat social, communication, and behavioral deficits and is known to modify the expression of synaptic receptors. We compared the effects of environmental enrichment in the expression of glutamate and endocannabinoid receptors, which are widely expressed in the cerebellar cortex. These two receptors interact to regulate neuronal function and their dysregulation is associated with behavioral changes. We used BTBR + Itpr3tf/J mice, a strain that models behavioral disorders, and C57BL/6 mice for comparison. We studied the effects of genetic background, sex, environmental conditions, and layer of the cerebellar cortex on the expression of each receptor. RESULTS: The influence of genetic background and environmental enrichment had the same pattern on glutamate and endocannabinoid receptors in males. In contrast, in females, the effect of environmental enrichment and genetic background were different than the ones obtained for males and were also different between the glutamate and endocannabinoid receptors. Furthermore, an analysis of both receptors from tissue obtained from the same animals show that their expression is correlated in males, but not in females. Our results suggest that environmental enrichment has a receptor dependent and sexual dimorphic effect on the molecular expression of different receptors in the cerebellar cortex.

OBJECTIVE: The combined use of action observation and motor imagery (AOMI) is a promising technique in neurorehabilitation that can be usefully applied in addition to conventional forms of therapy. Previous studies with healthy participants showed that the mere passive observation of walking results in a phase-dependent reflex modulation in the tibialis anterior muscle that resembles the pattern occurring when walking. In patients after stroke, a similar reflex modulation was found in several lower limb muscles during the real execution of walking, but responses were blunted. To clarify whether and how lower limb reflex responses are also modulated in such patients during the combined synchronous observation and imagery of walking, medium-latency cutaneous reflexes from the tibialis anterior muscle were measured. We compared the reflex responses of seven patients after stroke during the AOMI of walking from two different conditions: (a) elicited during the end stance phase and (b) during the end swing phase, both normalized to a baseline condition. RESULTS: So far, using the identical methodological set-up as in our study with healthy individuals, we could not find any noteworthy reflex response modulation. The study was registered with the German Clinical Trials Register (DRKS00028255). TRIAL REGISTRATION: The study was registered with the German Clinical Trials Register: DRKS00028255.

OBJECTIVES: We present a database on Brazilian spatial, demographic, and socioeconomic characteristics from 1996 to 2020. This database aims for integration and harmonization with epidemiological data from two major studies. It can also be a valuable database for designing and conducting various types of epidemiologic research, such as health inequality studies, ecological studies (mapping and time-trends), and multi-level analysis. DATA DESCRIPTION: The database gathers official information obtained via open sources from the Brazilian Institute of Geography and Statistics, the Institute for Applied Economic Research, and the Ministry of Health. It includes 139,153 observations and 26 attributes aggregated by years and policy-relevant geographic units on geocoding of municipality centroids, total population size, child population by age-group, birth and mortality measures, Brazilian Municipal Human Development Index, Gini coefficient, Gross Domestic Product, and sanitation. We automated all data processing and curation in the free and open software R.

OBJECTIVE: Aberrant expression in skeletal muscle of DUX4, a double homeobox transcription factor, underlies pathogenesis in facioscapulohumeral muscular dystrophy (FSHD). Although previous studies of FSHD muscle biopsies detected mRNAs encoding DUX4 and its target genes, no studies had reported detection of DUX4 protein. Our objective was to develop a proximity ligation assay (PLA) for DUX4 and to determine if this assay could detect DUX4 protein in FSHD muscle sections. RESULTS: We developed a PLA protocol using two DUX4 antibodies previously reported by Stephen Tapscott's group: P2G4, a mouse mAb specific for an epitope in the N-terminal region, and E5-5, a rabbit mAb specific for an epitope in the C-terminal region, in combination with commercial PLA secondary reagents. We validated the DUX4 PLA using cultured human myogenic cells in which DUX4 was ectopically expressed in a small fraction of nuclei. Using this two primary mAb PLA on an FSHD1 biceps biopsy, we observed nuclei with apparent DUX4 PLA signals associated with a small subset of myofibers (~ 0.05-0.1%). Though a limited pilot study, these results suggest that the two primary mAb PLA protocol could be useful for detecting DUX4 protein in FSHD muscle biopsies.

OBJECTIVES: 'Integrative and Conjugative Elements' (ICEs) and 'Integrative and Mobilizable Elements' (IMEs) are two classes of mobile genetic elements that are complex to detect and delineate. Therefore, they are yet poorly annotated in bacterial genomes. FirmiData provides to the scientific community of microbiologists and bioinformaticians a reference resource of annotated ICEs and of IMEs from Firmicutes. It illustrates their prevalence and their diversity but also gives information on their organization. FirmiData was designed to assist the scientific community in identifying and annotating these elements by using the sequences of these ICEs and IMEs for the identification of related elements in other genomes of Firmicutes. Therefore, Firmidata meets the needs of the scientific community. DATA DESCRIPTION: Firmidata provides a manually curated annotation of 98 ICEs and 148 IMEs identified in 40 chromosomes of Firmicutes. The delineation at the nucleotide level of almost all of these elements allows for the characterization of the genes they carry.

OBJECTIVE: The advent of new techniques such as video-assisted thoracoscopic surgery (VATS) for the removal of lung segments leads to compression of the surgical specimen, with the possible dissemination of neoplastic cells. The sheer volume of surgeries performed using these techniques has caused many institutions to stop removing the surgical specimen using an endobag, even when retractors/protectors are used in the instrumentalization incision. This study aimed to collect data from patients undergoing lung resection by VATS and analyze the cytopathological results of the collected material. RESULTS: A total of 47 endobag fluid samples were collected from patients who underwent VATS. The surgical specimen was subjected to histopathological analysis, and all patients underwent pathological TNM staging. In the cytopathological analyses, only 2 (4.3%) specimens of endobag fluid aspirate were positive for neoplastic cells. In these two cases, the tumors were peripheral, both with diagnoses of moderately differentiated adenocarcinoma and with classifications of T1bN0M0 and T3N0M0. These results indicate that although there is a low incidence of tumor cells in endobag fluid, it is always better to perform surgery using all available protective measures to avoid tumor implantation in the thoracic cavity to the greatest extent possible.

OBJECTIVE: The mesoderm-specific transcript (Mest) is an imprinted gene that is transcribed from the paternal allele. It is a marker of adipose tissue expansion; however, it is uncertain whether Mest expression promotes or suppresses adipogenic differentiation. To elucidate the effects of Mest expression on adipogenic differentiation, we transfected an expression vector or siRNA for mouse Mest into 3T3-L1 mouse preadipocyte cell line. RESULTS: In differentiated 3T3-L1 adipocytes, Mest overexpression decreased lipid accumulation. Conversely, gene silencing of Mest increased the accumulation of lipid droplets in adipocytes. These results demonstrate that Mest negatively regulates adipocyte differentiation. Further, Mest induced trans-differentiation of 3T3-L1 cells into hepatocytes, and its overexpression induced the expression of hepatocyte marker genes, including albumin and α-fetoprotein. In the presence of dexamethasone, the forced expression of the Mest caused morphological changes in 3T3-L1 cells. Cells were flat and polygonal shapes, with an increased accumulation of intracellular glycogen and other features that are typical of hepatocytes. Therefore, Mest inhibits adipogenic differentiation of 3T3-L1 preadipocytes by inducing hepatocyte trans-differentiation.

OBJECTIVES: The increasing spread of severe acute respiratory syndrome coronavirus-2 has prompted Canada to take unprecedented measures. The objective of this study was to examine the impact of the implemented public health measures on the incidence of COVID-19 in Manitoba. RESULTS: Using the COVID-19 dataset, we examined the temporal trends of daily reported COVID-19 cases and the coinciding public health measures implemented from March 12, 2020 to February 28, 2022. We calculated the 7-day moving average and crude COVID-19 infection rate/100,000 Manitobans. Due to the restrictions applied, the infection rate decreased from 2.4 (April 1) to 0.07 infections (May 1, 2020). Between May 4 and July 17, 2020, the reported cases stabilized, and some restrictions were lifted. However, in November, the cases peaked with infection rate of 29. Additional restrictions were implemented, and the rate dropped to 3.6 infections on March 31, 2021. As of August 2021, 62.8% of eligible Manitobans received two vaccine doses. The infection rate increased to 128.3 infections on December 31, 2021 and mitigation measures were implemented. This study describes how physical distancing in conjunction with other containment measures can reduce the COVID-19 burden. Future studies into the extent of the implementation of the restrictions are necessary.

OBJECTIVES: Research examining the age of diagnosis of autism spectrum disorder (ASD) and its influencing factors mostly originate from developed Western countries, providing little to no systematic information about the understanding and management of ASD in the rest of the world. The present exploratory study examined the influence of child and family characteristics on the age of ASD diagnosis in Saudi Arabia. RESULTS: The median age at diagnosis was 3.0 years and was associated with some child and family characteristics. A 1 year increase in child's age was associated with a 0.1 year increase in age of diagnosis (95% CI 0.05, 0.12). Children who did not respond to their name were diagnosed 0.3 years earlier than other children (95% CI - 0.60, - 0.05), and engaging in challenging behavior was associated with a 0.5 year increase in age of diagnosis (95% CI 0.20, 0.81). A lack of comorbidity was associated with a 0.6 year increase in the age of diagnosis compared to the diagnosis age of children with comorbidity (95% CI 0.13, 1.01). Finally, those residing outside of Saudi Arabia were diagnosed with ASD 0.9 years earlier than those residing in Saudi Arabia (95% CI - 0.171, - 0.11).

OBJECTIVE: The objective of this study was to identify the diagnostic performance of video capsule endoscopy (VCE) among patients presenting with iron deficiency anaemia (IDA) and negative bidirectional endoscopy to a gasteroendoscopy practice in regional Australia. The secondary objectives were to identify the distribution of findings and factors predictive of positive findings in a regional setting. RESULTS: In total 123 procedures were included in the study. Mean age of the patients was 67.9 years. Females made up 60.2% (n = 74) of the study population. Mean haemoglobin and ferritin levels were 93.3 g/L and 11.9 ug/L, respectively. Positive findings were present in 67 procedures (54.5%) with the most frequent finding being small bowel angiodysplasia (53.7%, n = 36/67), followed by ulceration/significant erosion (26.8%, n = 18/67), fresh blood (20.8%, n = 14/67) and tumour/polyp (16.4%, n = 11/67). Haemoglobin level was the only variable associated with positive findings (p = 0.005) in the study population. Of the procedures reporting positive findings outside the small bowel, the majority (80%) were within reach of conventional upper endoscopy and may have implications for future practice, particularly when allocating health resources in a rural setting.