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Abstract Background Insecticide resistance is jeopardising malaria control efforts in Africa. Deciphering the evolutionary dynamics of mosquito populations country-wide is essential for designing effective and sustainable national and subnational tailored strategies to accelerate malaria elimination efforts. Here, we employed genome-wide association studies through pooled template sequencing to compare four eco-geographically different populations of the major vector, Anopheles funestus, across a South North transect in Cameroon, aiming to identify genomic signatures of adaptive responses to insecticides. Results Our analysis revealed limited population structure within Northern and Central regions (F ST 0.049). Greater genetic differentiation was observed at known resistance-associated loci, resistance-to-pyrethroids 1 (rp1) (2R chromosome) and CYP9 (X chromosome), with varying signatures of positive selection across populations. Allelic variation between variants underscores the pervasive impact of selection pressures, with rp1 variants more prevalent in Central and Northern populations (F ST >0.3), and the CYP9 associated variants more pronounced in the Littoral/Coastal region (F ST =0.29). Evidence of selective sweeps was supported by negative Tajima’s D and reduced genetic diversity in all populations, particularly in Central (Elende) and Northern (Tibati) regions. Genomic variant analysis identified novel missense mutations and signatures of complex genomic alterations such as duplications, deletions, transposable element (TE) insertions, and chromosomal inversions, all associated with selective sweeps. A 4.3 kb TE insertion was fixed in all populations with Njombe Littoral/Coastal population, showing higher frequency of CYP9K1 (G454A), a known resistance allele and TE upstream compared to elsewhere. Conclusion Our study uncovered regional variations in insecticide resistance candidate variants, emphasizing the need for a streamlined DNA-based diagnostic assay for genomic surveillance across Africa. These findings will contribute to the development of tailored resistance management strategies crucial for addressing the dynamic challenges of malaria control in Cameroon.

Abstract Background Gaining a comprehensive understanding of the genetic mechanisms underlying insecticide resistance in malaria vectors is crucial for optimising the effectiveness of insecticide-based vector control methods and developing diagnostic tools for resistance management. Considering the heterogeneity of metabolic resistance in major malaria vectors, the implementation of tailored resistance management strategies is essential for successful vector control. Here, we provide evidence demonstrating that two highly selected mutations in CYP6P4a and CYP6P4b are driving pyrethroid insecticide resistance in the major malaria vector Anopheles funestus, in West Africa. Results Continent-wide polymorphism survey revealed escalated signatures of directional selection of both genes between 2014 and 2021. In vitro insecticide metabolism assays with recombinant enzymes from both genes showed that mutant alleles under selection exhibit higher metabolic efficiency than their wild-type counterparts. Using the GAL4-UAS expression system, transgenic Drosophila flies overexpressing mutant alleles exhibited increased resistance to pyrethroids. These findings were consistent with in silico predictions which highlighted changes in enzyme active site architecture that enhance the affinity of mutant alleles for type I and II pyrethroids. Furthermore, we designed two DNA-based assays for the detection of CYP6P4a-M220I and CYP6P4b-D284E mutations, showing their current confinement to West Africa. Genotype/phenotype correlation analyses revealed that these markers are strongly associated with resistance to types I and II pyrethroids and combine to drastically reduce killing effects of pyrethroid bed nets. Conclusions Overall, this study demonstrated that CYP6P4a and CYP6P4b contribute to pyrethroid resistance in An. funestus and provided two additional insecticide resistance molecular diagnostic tools that would contribute to monitoring and better management of resistance.

Abstract Objectives To implement and evaluate a super-fast and high-quality biparametric MRI (bpMRI) protocol for prostate imaging acquired at a new ultra-high gradient 3.0-T MRI system. Methods Participants with clinically suspected prostate cancer prospectively underwent a multiparametric MRI (mpMRI) on a new 3.0-T MRI scanner (maximum gradient strength: 200 mT/m, maximum slew rate: 200 T/m/s). The bpMRI protocol was extracted from the full mpMRI protocol, including axial T2-weighted and diffusion-weighted (DWI) sequences (b0/800, b1500). Overall image quality was rated by two readers on a five-point Likert scale from (1) non-diagnostic to (5) excellent. PI-RADS 2.1 scores were assessed by three readers separately for the bpMRI and mpMRI protocols. Cohen’s and Fleiss’ κ were calculated for PI-RADS agreement between protocols and interrater reliability between readers, respectively. Results Seventy-seven male participants (mean age, 66 ± 8 years) were included. Acquisition time of the bpMRI protocol was reduced by 62% (bpMRI: 5 min, 33 ± 21 s; mpMRI: 14 min, 50 ± 42 s). The bpMRI protocol showed excellent overall image quality for both the T2-weighted (median score both readers: 5 [IQR: 4–5]) and DWI (b1500) sequence (median score reader 1: 4 [IQR: 4–5]; reader 2: 4 [IQR: 4–4]). PI-RADS score agreement between protocols was excellent (Cohen’s κ range: 0.91–0.95 [95% CI: 0.89, 0.99]) with an overall good interrater reliability (Fleiss’ κ, 0.86 [95% CI: 0.80, 0.92]). Conclusion Ultra-high gradient MRI allows the establishment of a high-quality and rapidly acquired bpMRI with high PI-RADS agreement to a full mpMRI protocol. Trials registration Clinicaltrials.gov, NCT06244680, Registered 06 February 2024, retrospectively registered, https://classic.clinicaltrials.gov/ct2/show/NCT06244680 . Critical relevance statement A novel 3.0-Tesla MRI system with an ultra-high gradient performance enabled high-quality biparametric prostate MRI in 5.5 min while achieving excellent PI-RADS agreement with a standard multiparametric protocol. Key Points Multi- and biparametric prostate MRIs were prospectively acquired utilizing a maximum gradient of 200 mT/m. Super-fast biparametric MRIs showed excellent image quality and had high PI-RADS agreement with multiparametric MRIs. Implementation of high gradient MRI in clinical routine allows accelerated and high-quality biparametric prostate examinations. Graphical Abstract

Abstract Objectives To assess the prognostic value of cardiac MRI (CMR) parameters for the occurrence of major adverse cardiac events (MACE) in patients with infarct-like myocarditis. Methods In this retrospective single-center study, patients with CMR-confirmed acute myocarditis with infarct-like presentation were identified (2007–2020). Functional and structural parameters were analyzed including late gadolinium enhancement (LGE). The primary endpoint was the occurrence of MACE up to 5 years after discharge. Results In total, 130 patients (mean age, 40 ± 19 years; 97 men, 75%) with infarct-like myocarditis were included. CMR was conducted a median of 3 days (interquartile range [IQR], 1–5) after symptom onset. MACE occurred in 18/130 patients (14%) during a median follow-up of 19.3 months (IQR, 4.5–53). The median extent of LGE was 7% (IQR, 4–10). LGE affected the subepicardium in 111/130 patients (85%), the midwall in 45/130 patients (35%), and both the subepicardium and midwall in 27/130 patients (21%). Transmural extension of non-ischemic LGE lesions was observed in 15/130 patients (12%) and septal LGE in 42/130 patients (32%). In univariable Cox regression analysis, a significant association was found between the occurrence of MACE and both, quantified LGE extent and transmural LGE pattern. In multivariable analysis, transmural extension of LGE was an independent predictor for MACE (hazard ratio, 6.34; 95% confidence interval: 2.29–17.49; p

Abstract Background Higher educational attainment is important for economic wellbeing and associated with better health and longevity. Previous research focused on intelligence, socioeconomic status and mental health or individual risk behaviours as predictors of educational attainment, but the role of multiple domains of adolescent risk behaviours is less clear. This study examined the association between multiple domains of risk behaviour in adolescence and educational attainment by 22 years-of-age. Methods Young people (Generation 2, Gen2) and their parents (Generation 1, Gen1) participating in the Raine Study completed questionnaires at years 1, 5, 8, 10 (Gen1 only), 14, 17 (both) and 22 (Gen2 only). The Raine Study is an ongoing longitudinal study initiated in Perth, Western Australia, between 1989 and 1991. The 1,102 Gen2 participants who responded to questions about highest educational attainment were included in this study. The association between Gen2 self-reported risk behaviours (including age at commencement of drinking alcohol, smoking, sexual intercourse and drug use) and educational attainment (defined as self-reported years of completed high school: ≤10, 11, 12 or tertiary education (> 12)) at year 22, after adjusting for mother’s age and combined parental education level, participant sex, and family income, educational performance and adolescent mental health, was explored using ordinal regression models and presented as odds ratios (OR) with 95% confidence intervals (CI). Results Ordinal models suggested that never smoking or starting older than 18 compared with smoking before age 15 (OR 2.02, 95%CI: 1.28–2.14); first drinking alcohol between 15 and 17 years compared with younger than 15 (OR 1.52, 95%CI: 1.08–2.14); and, first sexual intercourse aged ≥ 18 years compared with under 15 (OR 1.67, 95%CI: 1.08–2.57) were associated with higher levels of educational attainment at 22-year follow-up. Additionally, lower (“better”) behavioural scores increased the odds of higher levels of attainment. Conclusions Absence of health risk behaviours at a younger age or later commencement was associated with higher educational attainment. Evidence-based interventions that address the societal influences underpinning risk behaviours in adolescents may support longer school retention.

Abstract Background Digital technology is ubiquitous in the lives of many children and parents. To better understand any influence of technology use by infants, and mothers, on child development, technology use should be considered within the broader family system context in which children develop. This study aimed to investigate associations between infant and maternal technology use and infant 12-month development, taking into consideration other family factors. Methods This cross-sectional study used data from ORIGINS participants, collected at 12-months of age: maternal and child technology use (TechU-Q), sociodemographic factors (e.g. child sex, household income), parental mental health (DASS-21), and child development (Ages and Stages Questionnaire). Linear regression was used for analyses. Results When family factors were considered, higher infant mobile touchscreen device (MTSD) use was associated with poorer infant development for gross motor, problem-solving, and total ASQ-3 scores. In contrast there were no associations between infant television (TV) watching or maternal technology use and total ASQ-3 scores. Higher maternal technology use was associated with higher infant technology use. Poorer maternal and paternal mental health were associated with poorer infant development. Poorer maternal mental health was also associated with higher infant TV watching and higher maternal MTSD use. Conclusion There is a complex relationship between technology use, parental mental health and other family factors that together influence infant development. To improve infant development outcomes, less focus should be on infant or maternal technology use, and more on supporting the family as a whole, and parental mental health in particular.

Two human induced pluripotent stem cell lines, LEIi021-A and LEIi021-B, were derived from dermal fibroblasts from a healthy control subject from an Australian Aboriginal family with retinitis pigmentosa-11. Reprogramming was performed using episomal vectors expressing OCT4, SOX2, LIN28, L-MYC, KLF4 and mp53DD. Pluripotency markers were expressed in both LEIi021-A and LEIi021-B lines. The two cell lines displayed normal karyotypes and demonstrated the ability to differentiate into embryoid bodies with the three primary germ layers and retinal pigment epithelial cells.

ABSTRACT The widespread prevalence of Asaia in mosquitoes makes it a potential candidate for paratrangenic control in Anopheles. To better understand whether this bacterium could be used for malaria control, we quantified Asaia in An. gambiae s.l populations in malaria endemic regions examining co-infection with Plasmodium falciparum. Adult Anopheles mosquitoes were collected across two different eco-geographical localities in Cameroon, during both the dry and wet seasons. DNA was extracted from whole individual mosquitoes, and real time-qPCR amplification of the 16S ribosomal RNA was used to quantify Asaia in both An. gambiae and An. coluzzii samples. We also detected and quantified P. falciparum infection in the same mosquitoes. The density of Asaia was successfully quantified in a total of 864 field mosquitoes, comprising of 439 An. gambiae from Bankeng and 424 An. coluzii collected from Gounougou. Interestingly, a higher prevalence of Asaia in An. gambiae (88.3%) compared to An. coluzzii (80.9%) was observed. Moreover, the density of Asaia in both species was significantly affected by seasonal changes in the two localities. Furthermore, a significant difference between the infection densities of Asaia and the Plasmodium infection status in the two species was recorded. However, no correlation was observed between the number of Asaia and P. falciparum infections. This study provides evidence that naturally occurring Asaia infection is not correlated to P. falciparum development within An. gambiae and An. coluzzii. Nevertheless, further studies incorporating experimental infections are required to better investigate the correlation between Anopheles mosquitoes, Asaia, and Plasmodium.IMPORTANCEThe symbiont Asaia has emerged as a promising candidate for paratransgenic control of malaria, but further analysis of its biology and genetics across Africa is necessary. In this study, we investigated and quantified the influence of Asaia in naturally infected An. gambiae s.l. populations with the malaria parasite Plasmodium falciparum. Genomic DNA was extracted from whole individual mosquitoes collected from two localities, and Asaia was quantified using real-time qPCR by amplification of the 16S ribosomal RNA gene. We also detected and quantified Plasmodium falciparum infection in the same mosquitoes and established the correlation between Asaia and Plasmodium coinfection. This study provides evidence that naturally occurring Asaia infection is not correlated with P. falciparum development within An. gambiae and An. coluzzii mosquitoes.

Background We recently found that loss of endothelial cell APOE3 disrupts neurovascular and synaptic function. However, whether endothelial APOE4 is detrimental or protective for neural function under physiological conditions is unknown. Therefore, the goal of this study was to determine the role of endothelial cell APOE4 in regulating brain function in vivo. Methods and Results We developed APOE4fl/fl/Cdh5(PAC)‐CreERT2+/− and APOE4fl/fl/Cdh5(PAC)‐CreERT2−/− (control) mice. Knockdown of endothelial cell APOE4 was induced at ≈4 to 5 weeks of age. Experiments were conducted at 9 months of age to evaluate neurovascular and neuronal function via biochemistry, immunohistochemistry, behavior tests, and electrophysiology. Endothelial cell APOE4 knockdown resulted in higher neurovascular permeability, lower claudin‐5 vessel coverage, impaired trace fear memory extinction, and disruption of cortical excitatory‐inhibitory balance of synaptic activity. Conclusions Our data support the novel concept that endothelial cell APOE4 is protective for brain function when other cell types express APOE4.

Most of the short-chain fatty acids (SCFAs) are produced by Bifidobacterium, Lactobacillus, Lachnospiraceae, Blautia, Coprococcus, Roseburia, Facealibacterium and Oscillospira. Butyrate (C4H7O2−) supplies 70% of energy to intestinal epithelial cells (IECs), supports tight-junction protein formation, induces the production of inflammatory cytokines, and inhibits histone deacetylase (HDAC). Butyrate is also associated with the recovery of brain trauma, improvement of dementia, the alleviation of autoimmune encephalitis, and several intestinal disorders. Low levels of SCFAs are associated with hypertension, cardiovascular disease (CVD), strokes, obesity, and diabetes mellitus. Cis-palmitoleic acid (C16H30O2), a mono-unsaturated fatty acid (MUFA), increases insulin sensitivity and reduces the risk of developing CVD. Lipokine palmitoleic acid reduces the expression of pro-inflammatory cytokines IL-1β (pro-IL1β), tumor necrosis factor α (TNF-α), and isoleucine 6 (IL-6). Polyunsaturated fatty acids (PUFAs), such as omega-3 and omega-6, are supplied through the diet. The conversion of PUFAs by cyclooxygenases (COX) and lipoxygenases (LOX) leads to the production of anti-inflammatory prostaglandins and leukotrienes. Oxidation of linoleic acid (LA, C18H32O2), an omega-6 essential fatty acid, leads to the formation of 13-hydroperoxy octadecadienoic acid (13-HPODE, C18H32O4), which induces pro-inflammatory cytokines. Omega-3 PUFAs, such as eicosapentaenoic acid (EPA, C20H30O2) and docosahexaenoic acid (DHA, C22H32O2), lower triglyceride levels, lower the risk of developing some sort of cancers, Alzheimer’s disease and dementia. In this review, the importance of SCFAs, MUFAs, PUFAs, and saturated fatty acids (SFAs) on human health is discussed. The use of fatty acids in the treatment of diseases is investigated.

Background The acquisition of contrast‐enhanced T1 maps to calculate extracellular volume (ECV) requires contrast agent administration and is time consuming. This study investigates generative adversarial networks for contrast‐free, virtual extracellular volume (vECV) by generating virtual contrast‐enhanced T1 maps. Methods and Results This retrospective study includes 2518 registered native and contrast‐enhanced T1 maps from 1000 patients who underwent cardiovascular magnetic resonance at 1.5 Tesla. Recent hematocrit values of 123 patients (hold‐out test) and 96 patients from a different institution (external evaluation) allowed for calculation of conventional ECV. A generative adversarial network was trained to generate virtual contrast‐enhanced T1 maps from native T1 maps for vECV creation. Mean and SD of the difference per patient (ΔECV) were calculated and compared by permutation of the 2‐sided t test with 10 000 resamples. For ECV and vECV, differences in area under the receiver operating characteristic curve (AUC) for discriminating hold‐out test patients with normal cardiovascular magnetic resonance versus myocarditis or amyloidosis were tested with Delong's test. ECV and vECV showed a high agreement in patients with myocarditis (ΔECV: hold‐out test, 2.0%±1.5%; external evaluation, 1.9%±1.7%) and normal cardiovascular magnetic resonance (ΔECV: hold‐out test, 1.9%±1.4%; external evaluation, 1.5%±1.2%), but variations in amyloidosis were higher (ΔECV: hold‐out test, 6.2%±6.0%; external evaluation, 15.5%±6.4%). In the hold‐out test, ECV and vECV had a comparable AUC for the diagnosis of myocarditis (ECV AUC, 0.77 versus vECV AUC, 0.76; P=0.76) and amyloidosis (ECV AUC, 0.99 versus vECV AUC, 0.96; P=0.52). Conclusions Generation of vECV on the basis of native T1 maps is feasible. Multicenter training data are required to further enhance generalizability of vECV in amyloidosis.