High-throughput sequencing provides sufficient means for determining genotypes of clinically important pharmacogenes that can be used to tailor medical decisions to individual patients. However, pharmacogene genotyping, also known as star-allele calling, is a challenging problem that requires accurate copy number calling, structural variation identification, variant calling, and phasing within each pharmacogene copy present in the sample. Here we introduce Aldy 4, a fast and efficient tool for genotyping pharmacogenes that uses combinatorial optimization for accurate star-allele calling across different sequencing technologies. Aldy 4 adds support for long reads and uses a novel phasing model and improved copy number and variant calling models. We compare Aldy 4 against the current state-of-the-art star-allele callers on a large and diverse set of samples and genes sequenced by various sequencing technologies, such as whole-genome and targeted Illumina sequencing, barcoded 10x Genomics, and Pacific Biosciences (PacBio) HiFi. We show that Aldy 4 is the most accurate star-allele caller with near-perfect accuracy in all evaluated contexts, and hope that Aldy remains an invaluable tool in the clinical toolbox even with the advent of long-read sequencing technologies.

Background: Response to the outbreak of poliomyelitis in mid-1950 led to recognition and consequent development of critical care. Seventy years later the humankind was struck by COVID-19, another major challenge for critical care medicine which was especially big in Low-Resources-Settings where more than two thirds of the world population live, including the population of the Republic of Srpska (RS). Design and methods: The main aim was to show an overview of all interventions in order to boost hospitals’ capacities to the level which is sufficient to manage high amount of critically ill COVID-19 patients in the RS. A before-after cohort study design was conducted to evaluate the effects of interventions for increase in preparedness and capacity of ICUs for admission and treatment of COVID-19 critically ill patients in nine hospitals in the RS. Results: Following interventions, the biggest and university affiliated hospital in the RS has increased ICU capacities: total number of ICU beds increased by 38% and number of ventilators by 114%. Availability of machines for veno-venous extracorporeal membrane oxygenation (vvECMO) increased by 100%. Number of doctors who were involved in treatment of critically ill patients increased by 47% and nurse/patient’s ratio reached 1:2.5. Similarly, all other hospitals experienced boosting of ICU beds by 189% and ventilators by 373% while number of doctors increased by 108% and nurse/patient’s ratio reached 1:4. Conclusion: All interventions implemented during COVID-19 pandemic outbreak in the RS resulted in increasing capacity for treatment of critically ill patients, but the education of health care professionals was identified as the most important conducted intervention.

Purpose Paediatric Type 1 Diabetes (T1D) patients are at greater risk for developing severe hypo and hyperglycaemic events due to poor glycaemic control. To reduce the risk of adverse events, patients need to achieve the best possible glycaemic management through frequent blood glucose monitoring with finger prick or Continuous Glucose Monitoring (CGM) systems. However, several non-invasive techniques have been proposed aiming at exploiting changes in physiological parameters based on glucose levels. The overall objective of this study is to validate an artificial intelligence (AI) based algorithm to detect glycaemic events using ECG signals collected through non-invasive device. Methods This study will enrol T1D paediatric participants who already use CGM. Participants will wear an additional non-invasive wearable device for recording physiological data and respiratory rate. Glycaemic measurements driven through ECG variables are the main outcomes. Data collected will be used to design, develop and validate the personalised and generalized classifiers based on a deep learning (DL) AI algorithm, able to automatically detect hypoglycaemic events by using few ECG heartbeats recorded with wearable devices. Results Data collection is expected to be completed approximately by June 2023. It is expected that sufficient data will be collected to develop and validate the AI algorithm. Conclusion This is a validation study that will perform additional tests on a larger diabetes sample population to validate the previous pilot results that were based on four healthy adults, providing evidence on the reliability of the AI algorithm in detecting glycaemic events in paediatric diabetic patients in free-living conditions. Trial registration ClinicalTrials.gov identifier: NCT03936634. Registered on 11 March 2022, retrospectively registered, https://www.clinicaltrials.gov/ct2/show/NCT05278143?titles=AI+for+Glycemic+Events+Detection+Via+ECG+in+a+Pediatric+Population&draw=2&rank=1 .

Artemisia annua L. has long been known for its medicinal properties and isolation of ingredients whose derivatives are used for therapeutic purposes. The CYP2B6 and CYP3A4 enzymes belong to a large family of cytochrome P450 enzymes. These enzymes are involved in the metabolism of drugs and other xeonobiotics. It is known that various compounds can induce or inhibit the activity of these enzymes. The aim of this study was to investigate the nature of the inhibitory effect of Artemisia annua extract on CYP2B6 and CYP3A4 enzymes, as well as the type of inhibition, the presence of reversible or pseudo-irreversible inhibition, and the possible heme destruction. The methanolic extract of Artemisia annua showed an inhibitory effect on CYP2B6 (by almost 90%) and CYP3A4 enzymes (by almost 70%). A significant decrease in heme concentration by 46.8% and 38.2% was observed in different assays. These results clearly indicate that the studied plant extracts significantly inhibited the activity of CYP2B6 and CYP3A4 enzymes. Moreover, they showed irreversible inhibition, which is even more important for possible interactions with drugs and dietary supplements.

With the goal of enhancing the quality of the environment, urban green infrastructure (UGI) is an essential element in sustainable cities, and nature-based solutions (NBS) are being carried out as new infrastructure solutions that increase the resilience of cities. In this research, the method of theoretical analysis and the content analysis as the basic fact-gathering technique was applied to answer to following questions: What are the hindrances and bottlenecks in implementing NBS? Are the current decision-making mechanisms helping NBS get in route to shape cities? Is there any binding policy in practice that promotes NBS? In Belgrade is planned Type 3 of the degree of intervention/level and engineering type—Creation and new ecosystem management in the classifications of intensive urban green space management; urban planning strategies; urban water management; ecological restoration of degraded terrestrial ecosystems; and restoration and creation of semi-natural water bodies and hydrographic networks. In the future, it is essential to implement policies and incentives on national, regional, and local scales that help encourage the usage of NBS in the development of urban infrastructure.

Purpose: Total elbow arthroplasty (TEA) is rarely performed compared to other arthroplasties. For many surgical procedures, literature shows better outcomes when they are performed by experienced surgeons and in so-called ‘high-volume’ hospitals. We systematically reviewed the literature on the relationship between surgical volume and outcomes following TEA. Methods: A literature search was performed using the MEDLINE, EMBASE and CINAHL databases. The literature was systematically reviewed for original studies comparing TEA outcomes among hospitals or surgeons with different annual or career volumes. For each study, data were collected on study design, indications for TEA, number of included patients, implant types, cut-off values for volume, number and types of complications, revision rate and functional outcome measures. The methodological quality of the included studies was assessed using the Newcastle–Ottawa Scale. Results: Two studies, which included a combined 2301 TEAs, found that higher surgeon volumes were associated with lower revision rates. The examined complication rates did not differ between high- and low-volume surgeons. In one study, low-hospital volume is associated with an increased risk of revision compared to high-volume hospitals, but for other complication types, no difference was found. Conclusions: Based on the results, the evidence suggests that high-volume centers have a lower revision rate in the long term. No minimum amount of procedures per year can be advised, as the included studies have different cut-off values between groups. As higher surgeon- and center-volume, (therefore presumably experience) appear to yield better outcomes, centralization of total elbow arthroplasty should be encouraged.

The emergence of SARS-CoV-2 variants of concern (VOC) is driven by mutations that mediate escape from neutralizing antibodies. There is also evidence that mutations can cause loss of T cell epitopes. However, studies on viral escape from T cell immunity have been hampered by uncertain estimates of epitope prevalence. Here, we map and quantify CD8 T cell responses to SARS-CoV-2-specific minimal epitopes in blood drawn from April to June 2020 from 83 COVID-19 convalescents. Among 37 HLA ligands eluted from five prevalent alleles and an additional 86 predicted binders, we identify 29 epitopes with an immunoprevalence ranging from 3% to 100% among individuals expressing the relevant HLA allele. Mutations in VOC are reported in 10.3% of the epitopes, while 20.6% of the non-immunogenic peptides are mutated in VOC. The nine most prevalent epitopes are conserved in VOC. Thus, comprehensive mapping of epitope prevalence does not provide evidence that mutations in VOC are driven by escape of T cell immunity.

Ancient Roman concretes have survived millennia, but mechanistic insights into their durability remain an enigma. Here, we use a multiscale correlative elemental and chemical mapping approach to investigating relict lime clasts, a ubiquitous and conspicuous mineral component associated with ancient Roman mortars. Together, these analyses provide new insights into mortar preparation methodologies and provide evidence that the Romans employed hot mixing, using quicklime in conjunction with, or instead of, slaked lime, to create an environment where high surface area aggregate-scale lime clasts are retained within the mortar matrix. Inspired by these findings, we propose that these macroscopic inclusions might serve as critical sources of reactive calcium for long-term pore and crack-filling or post-pozzolanic reactivity within the cementitious constructs. The subsequent development and testing of modern lime clast–containing cementitious mixtures demonstrate their self-healing potential, thus paving the way for the development of more durable, resilient, and sustainable concrete formulations.

Background Clinical guidelines recommend the identification of asthma comorbidities, especially treatable problems such as parental behaviours and child and parent anxiety. Purpose We aimed to (1) explore associations of asthma severity with child and parent state anxiety, trait anxiety, and asthma-related anxiety as well as with caregiver behaviours around physical activity and (2) explore if caregiver behaviours around physical activity were associated with use of inhaled β-Agonists when symptomatic, and with child and parent anxiety. Patients and methods Patients ages 3–17 years with asthma (n = 72) and their parents were recruited from the Pulmonology-Allergology Pediatric clinic University Hospital Centre Split in Split, Croatia during 2021. During a clinical visit, the pharmacological regimen was assessed and spirometry was performed. Children completed the State-Trait Anxiety Inventory for Children (STAI-C) and the Youth Asthma-Related Anxiety Scale (YASS). Parents completed the State-Trait Anxiety Inventory (STAI), the Parent Asthma-Related Anxiety Scale (PASS), and the Physical Activity Parenting Practices – Short Form (PAPP). Results Most patients had mild asthma (69.4%). Children with moderate to severe asthma had increased asthma-related anxiety (mean = 11.94 ± 6.1) compared with children with mild asthma (mean = 5.97 ± 6.39, p = 0.003). Parents of children with mild asthma reported behaviours allowing unsupervised physical activity outside more often when compared to parents of children with moderate or severe asthma. Physical activity facilitation parenting behaviour reduced the odds of a child's need for quick-reliever medication when symptomatic (OR = 0.376,95% CI = −1.885 to −0.072; p = 0.034); more coercive parenting increased the odds of a child's additional use of such medications (OR = 2.602; 95% CI = 0.005 to 1.908; p = 0.049). Parents of children in the highest quartile of trait anxiety showed less non-directive support (1.97 ± 1.01 vs. 2.89 ± 1.19, p = 0.031) and less autonomy support (3.14 ± 1.32 vs. 4.11 ± 1.23, p = 0.037) of physical activity in their children than those with less trait anxiety. Conclusion Asthma-related anxiety was an important construct in this sample of children, associated with their disease severity as well as their parent's behaviours around the child's physical activities. Current research, conducted during the COVID-19 pandemic, recognised the tangible ways that parents support or avoid the asthmatic children's physical activity participation. Child anxiety and recognised parental physical activity behaviours are potentially important factors to assess and target for intervention.

Abstract Biotic stress caused by pests on citrus trees has become a major problem for ecologists around the world, hence the need to develop ecological approaches to surmount this stress. To evaluate the effectiveness of diverse control methods on the pest Panonychus citri and its predators, Euseius stipulatus, Phytoseiulus persimilis, and Typhlodromus sp. Different treatments were applied in orchards planted by Citrus clementina in the Gharb area of Morocco (T0 = treated by water only, T1 = spirodiclofen (the molecular formula is C21H24Cl2O4). 0.5 L/ha, T2 = 5% black soap 125 L/ha, Its ingredients are: argan oil, olive oil, black olive, salt, water, vegetable potash, without chemicals. T3 = mixture of two detergents; Oni product is composed of the following ingredients (sodium C12-13 parenth sulfate, and sodium C14-17 alkyl sec sulfonate) and Tide product with the following ingredients (sodium C10-16 alkylbenzene sulfonate, propylene glycol and sodium borate). Results obtained during the follow-up period showed that the black soap, spirodiclofen, and two detergent treatments provided a decrease in the P. citri population in comparison with plot treated by water during the whole monitoring period. We found 5,044 mobile forms of P. citri in the plot treated by water only, 1,544 forms in the plot treated with spirodiclofen, 3,002 forms in the plot treated with black soap, and 1,567 mobile forms in the plot treated with two detergents. This study demonstrated that all three treatments had a negative effect on the predators that survived in comparison to the control (T0), while the negative impact of spirodiclofen on beneficial phytoseiid mites was greater than that of black soap and detergents. However, the increase in temperature according to the follow-up weeks showed a significant effect on the evolution of the density of the studied mites. Unfortunately, the chemical products like spirodiclofen used against various citrus pests are considered toxic to phytoseiids and to the agro-ecosystem in general. This comparison aims to detect the effectiveness of black soap and mixtures of two detergents on P. citri and their impacts on its predators: E. stipulatus, P. persimilis, and Typhlodromus sp. to replace the use of pesticides by these alternative methods, which remain effective and environmentally friendly.