Improving Sphenophorus levis Adult Mortality Through Solid Insecticide Applications and Increased Insecticide Dose.

The sugarcane weevil (Sphenophorus levis Vaurie, 1978) is currently considered the most important sugarcane pest in Brazil, causing significant yield losses. Application methods of insecticides for S. levis control have not been effective, mostly due to the insect's cryptic behavior below the soil surface which suppresses the correct placement of insecticide active ingredients on target. Two experiments were conducted using an innovative bioassay methodology that simulates sugarcane field conditions to effectively evaluate S. levis adult mortality and insecticide residues in the soil under different treatments. The first experiment aimed to assess the efficacy of two liquid- and solid-applied insecticides, while the second aimed to examine the effect of increasing the dose of lambda-cyhalothrin + thiamethoxam on S. levis adult control. The novel bioassays simulated liquid and solid insecticide applications on sugarcane by exposing S. levis adults to chemical residuals on rhizomes and in soil after insecticide application. In the first experiment, low S. levis adult control was detected (< 53% mortality) across all treatments, where both solid and liquid applications of lambda-cyhalothrin + thiamethoxam provided greater efficacy levels than imidacloprid and control treatments, respectively. Solid applications maintained higher insecticide concentrations in the soil for longer periods than liquid insecticide applications, providing maximum insect control levels 7 days after application. The second experiment revealed that solid applications at higher insecticide doses significantly improved control of S. levis adult (76.7% mortality) and resulted in greater insecticide concentrations in the soil compared to the recommended label rate (58.8% mortality).

Brain-first forms of Parkinson's disease are over-represented in patients with non-responsive resting tremor.

Motor subtypes in Parkinson's Disease (PD) are unstable over time, limiting mechanistic insights and biomarker discovery. We focused on Rest Tremor (RT) as a symptom to test for phenotype stability and link it to specific circuits and disease mechanisms. Using the PPMI cohort data over 5 years we found that RT is more stable than classical Tremor-Dominant definitions, a stability also seen for RT response to therapy. At time of diagnosis, the population of therapy-resistant RT patients was enriched with a brain-first PD profile as predicted by a-Synuclein origin site and connectome (SOC) model. Resistant-RT patients have lower gastrointestinal and cardiovascular symptoms, lower prevalence of probable REM-Sleep behaviour disorder, and higher dopaminergic asymmetry compared to therapy-responsive or no tremor patients. Treating RT as a distinct phenomenon revealed a relative phenotypic stability with treatment response being linked to different patterns of disease progression.

Aslanger's pattern with acute lesion in the left coronary system: A case report.

Acute Coronary Syndrome (ACS) is characterized by the suspicion or confirmation of acute ischemia or acute myocardial infarction (MI). The presence of ST segment elevation (STE) consists in the main criterion for indication of immediate reperfusion therapy due to acute coronary occlusion, although significant part of the acute coronary occlusion cases do not fit the ST-segment elevation myocardial infarction (STEMI) criteria. A case report of a 50 year-old male presenting typical anginal pain, whose electrocardiogram (EKG) presented Aslanger's pattern. The patient had severe lesion of the left anterior descending artery (ADA) and chronic occlusion of the right coronary artery on cardiac catheterization, with important collateral flow from branches of the left coronary artery to the right coronary artery. The dichotomy between STEMI and Non-ST segment elevation myocardial infarction (NSTEMI) must be contested, owing to the evidence that one third of the NSTEMI patients presents a total coronary occlusion. There are other electrocardiographic patterns which must be considered to diagnose ACS. Our case report outlines a different clinical presentation of Aslanger pattern, that shows a ADA acute occlusion, instead of the lesion being in the right coronary system. There is evidence that, in addition to the STEMI vs NSTEMI criteria, the Aslanger pattern and other electrocardiographic patterns characterize occlusive ACS. Recognizing those patterns in clinical practice is essential to improve the diagnosis and early treatment of patients with ACS.

Inactivation in vitro of the marine parasite Amyloodinium ocellatum.

The ectoparasite Amyloodinium ocellatum is a dinoflagellate that causes severe morbidity and mortality in both brackish and marine warmwater aquaculture fishes worldwide. A. ocellatum has a triphasic life cycle based on a free-living flagellate (the dinospore), a parasitic stage (the trophont) and a resting and reproductive cyst (the tomont). Current chemical treatments have shown some efficacy in eliminating dinospores but fail to inactivate the tomonts. Here we evaluated the efficacy of alternative treatments in vitro through sporulation tests and the quantification of dinospore production and motility. Hydrogen peroxide and peracetic acid efficiently decreased dinospore production at low concentrations, but total inactivation of tomonts was only achieved with higher dosages. Tomont inactivation was also observed with disinfectants such as sodium hypochlorite and Virkon S. This work provides insights into effective and environmentally friendly alternatives for the elimination of resistant forms of the marine parasite A. ocellatum.

Asparagopsis taxiformis as a Novel Antioxidant Ingredient for Climate-Smart Aquaculture: Antioxidant, Metabolic and Digestive Modulation in Juvenile White Seabream (Diplodus sargus) Exposed to a Marine Heatwave.

The increasing frequency and duration of marine heatwaves (MHWs) due to climate change pose severe threats to aquaculture, causing drastic physiological and growth impairments in farmed fish, undermining their resilience against additional environmental pressures. To ensure sustainable production that meets the global seafood demand and animal welfare standards, cost-effective and eco-friendly strategies are urgently needed. This study explored the efficacy of the red macroalga Asparagopsis taxiformis on juvenile white seabream Diplodus sargus reared under optimal conditions and upon exposure to a MHW. Fish were fed with four experimental diets (0%, 1.5%, 3% or 6% of dried powdered A. taxiformis) for a prophylactic period of 30 days (T30) and subsequently exposed to a Mediterranean category II MHW for 15 days (T53). Biometric data and samples were collected at T30, T53 and T61 (8 days post-MHW recovery), to assess performance indicators, biomarker responses and histopathological alterations. Results showed that A. taxiformis supplementation improved catalase and glutathione S-transferase activities and reduced lipid peroxidation promoted by the MHW, particularly in fish biofortified with 1.5% inclusion level. No histopathological alterations were observed after 30 days. Additionally, fish biofortified with 1.5% A. taxiformis exhibited increased citrate synthase activity and fish supplemented with 1.5% and 3% showed improved digestive enzyme activities (e.g., pepsin and trypsin activities). Overall, the present findings pointed to 1.5% inclusion as the optimal dosage for aquafeeds biofortification with A. taxiformis, and confirmed that this seaweed species is a promising cost-effective ingredient with functional properties and great potential for usage in a climate-smart context.

Unraveling the Impact of miR-146a in Pulmonary Arterial Hypertension Pathophysiology and Right Ventricular Function.

Pulmonary arterial hypertension (PAH) is a chronic disorder characterized by excessive pulmonary vascular remodeling, leading to elevated pulmonary vascular resistance and right ventricle (RV) overload and failure. MicroRNA-146a (miR-146a) promotes vascular smooth muscle cell proliferation and vascular neointimal hyperplasia, both hallmarks of PAH. This study aimed to investigate the effects of miR-146a through pharmacological or genetic inhibition on experimental PAH and RV pressure overload animal models. Additionally, we examined the overexpression of miR-146a on human pulmonary artery smooth muscle cells (hPASMCs). Here, we showed that miR-146a genic expression was increased in the lungs of patients with PAH and the plasma of monocrotaline (MCT) rats. Interestingly, genetic ablation of miR-146a improved RV hypertrophy and systolic pressures in Sugen 5415/hypoxia (SuHx) and pulmonary arterial banding (PAB) mice. Pharmacological inhibition of miR-146a improved RV remodeling in PAB-wild type mice and MCT rats, and enhanced exercise capacity in MCT rats. However, overexpression of miR-146a did not affect proliferation, migration, and apoptosis in control-hPASMCs. Our findings show that miR-146a may play a significant role in RV function and remodeling, representing a promising therapeutic target for RV hypertrophy and, consequently, PAH.

SCI1, a flower regulator of cell proliferation, and its partners NtCDKG2 and NtRH35 interact with the splicing machinery.

Successful plant reproduction depends on the adequate development of flower organs controlled by cell proliferation and other processes. The SCI1 gene regulates cell proliferation and affects the final size of the female reproductive organ. To unravel the molecular mechanism exerted by SCI1 in cell proliferation control, we searched for its interaction partners through semi-in vivo pulldown experiments, uncovering a cyclin-dependent kinase, NtCDKG;2. Bimolecular fluorescence complementation (BiFC) and co-localization experiments showed that SCI1 interacts with NtCDKG;2 and its cognate NtCyclin L in nucleoli and splicing speckles. The screening of a yeast two-hybrid (Y2H) cDNA library using SCI1 as bait revealed a novel DEAD-box RNA helicase (NtRH35). The interaction between the NtCDKG;2-NtCyclin L complex, and NtRH35 was also shown. Subcellular localization experiments showed that SCI1, NtRH35, and the NtCDKG;2-NtCyclin L complex associate with each other within splicing speckles. The Y2H screening of NtCDKG;2 and NtRH35 identified the conserved spliceosome components U2a', NKAP, and CACTIN. This work presents SCI1 and its interactors NtCDKG;2-NtCyclin L complex, and NtRH35 as new spliceosome-associated proteins. Our findings reveal a network of interactions and suggest that SCI1 may regulate cell proliferation through the splicing process. This study provides new valuable insights into the intricate molecular pathways governing plant development.

Exploring the Influence of Date Palm Cultivars on Soil Microbiota.

Plants thrive in diverse environments, where root-microbe interactions play a pivotal role. Date palm (Phoenix dactylifera L.), with its genetic diversity and resilience, is an ideal model for studying microbial adaptation to different genotypes and stresses. This study aimed to analyze the bacterial and fungal communities associated with traditional date palm cultivars and the widely cultivated "Deglet Nour" were explored using metabarcoding approaches. The microbial diversity analysis identified a rich community with 13,189 bacterial and 6442 fungal Amplicon Sequence Variants (ASVs). Actinobacteriota, Proteobacteria, and Bacteroidota dominated bacterial communities, while Ascomycota dominated fungal communities. Analysis of the microbial community revealed the emergence of two distinct clusters correlating with specific date palm cultivars, but fungal communities showed higher sensitivity to date palm genotype variations compared to bacterial communities. The commercial cultivar "Deglet Nour" exhibited a unique microbial composition enriched in pathogenic fungal taxa, which was correlated with its genetic distance. Overall, our study contributes to understanding the complex interactions between date palm genotypes and soil microbiota, highlighting the genotype role in microbial community structure, particularly among fungi. These findings suggest correlations between date palm genotype, stress tolerance, and microbial assembly, with implications for plant health and resilience. Further research is needed to elucidate genotype-specific microbial interactions and their role in enhancing plant resilience to environmental stresses.

Role of myocardial microRNAs in the long-term ventricular remodelling of patients with aortic stenosis.

Aims: We hypothesize that miRs are key players in the dynamics of the hypertrophy phenotype in aortic stenosis (AS) patients. In our study, we aimed to identify the transcriptional patterns (protein-coding transcripts and miRs) from myocardial sample biopsies that could be associated with the absence of left ventricular (LV) mass regression after aortic valve replacement (AVR) in patients with severe AS and LV hypertrophy. Methods and results: We prospectively included 40 patients with severe AS, LV hypertrophy, and preserved ejection fraction undergoing AVR. Myocardial biopsies obtained during surgery were analysed for transcriptomic analysis performed by next-generation sequencing. At a 1-year follow-up, no hypertrophy reversal was observed in about half of the patients in the absence of patient-prosthesis mismatch and prosthesis dysfunction of uncontrolled hypertension. Predictors of mass regression were assessed from clinical, echocardiographic, and biochemical variables as well as from 300 miRs obtained from myocardial specimens, allowing the identification 29 differentially expressed. miR-4709-3p was found as a positive independent predictor of hypertrophy regression together with high-sensitivity troponin T (cTNT-hs) as a negative predictor. Gene transcripts RFX1, SIX5, MAPK8IF3, and PKD1 were predicted as simultaneous targets of five upregulated miRs suggesting its importance in LV hypertrophy. Conclusion: In our cohort, tissue miR-4709-3p and cTNT-hs were independent predictors of hypertrophy regression. The hypertrophy reversal process will likely depend from a complex network where miRNAs may have an important role, allowing a potential opportunity for therapy.

Eco-innovative aquafeeds biofortified with Asparagopsis taxiformis to improve the resilience of farmed white seabream (Diplodus sargus) to marine heatwave events.

Extreme weather events, like marine heatwaves (MHWs), are becoming more frequent and severe due to climate change, posing several challenges to marine ecosystems and their services. As disease outbreaks are often prompted by these acute phenomena, it is essential to develop eco-innovative strategies that can efficiently improve farmed fish resilience, especially under sub-optimal rearing conditions, thereby ensuring a sustainable aquaculture production. This study aimed to unveil farmed juvenile white seabream (Diplodus sargus, 28.50 ± 1.10 g weight, n = 150) immune and antioxidant responses under a category II MHW in the Mediterranean Sea (+4 °C, 8 days of temperature increase plus 15 days of plateau at the peak temperature) and to investigate whether a 30 days period of prophylactic biofortification with Asparagopsis taxiformis (1.5 %, 3 % and 6 %) enhanced fish resilience to these extreme events. Several biomarkers from different organization levels (individual, cellular, biochemical and molecular) were assessed upon 30 days of biofortification (T30), exposure (after 8 days of temperature increase + 15 days at peak temperature, T53) and recovery (8 days of temperature decrease, T61) from the MHW. Results showed that MHW negatively affected the fish physiological status and overall well-being, decreasing specific growth rate (SGR) and haematocrit (Ht) and increasing erythrocyte nuclear abnormalities (ENAs) and lipid peroxidation (LPO). These adverse effects were alleviated through biofortification with A. taxiformis. Seaweed inclusion at 1.5 % was the most effective dose to minimize the severity of MHW effects, significantly improving immune responses of D. sargus (i.e. increased levels of immunoglobulin M, peroxidase activity and lysozyme expression) and modulating antioxidant responses (i.e. decreased LPO, catalase and glutathione S-transferase activity). These findings confirm that A. taxiformis is a functional ingredient of added value to the aquaculture industry, as its inclusion in marine fish diets can beneficially modulate fish immunity and resilience under optimal and adverse rearing conditions.

Quality of Informed Consent in Phase III Clinical Trials in Portugal: The Participants' Perspective.

INTRODUCTION: Some studies show that participants do not always fully understand the informed consent form (ICF), which is one of the reasons for dropouts. This study aimed to adapt the Quality of Informed Consent (QuIC) questionnaire into a valid instrument to be applied to the Portuguese population and to measure its reliability and validity in the Portuguese population, by applying it to a sample of participants in controlled trials. METHODS: The QuIC questionnaire was developed to assess the quality of informed consent in clinical trials and consists of two parts, addressing both the objective (part A) and the subjective (part B) understanding. After being translated and validated into Portuguese, it was implemented in 100 cardiac participants of phase III clinical trials in a University Hospital Center. RESULTS: The QuIC-PT questionnaire showed excellent stability over time and good validity. All patients evaluated their participation and their health positively and recognized the main purpose of the clinical trial. Almost all participants understood their role in helping future patients and the purpose of the trial and realized that, by signing the ICF, they were participating in a clinical trial. However, none of them knew that their experimental treatment was not proven to be the best alternative for their condition. CONCLUSION: The QuIC-PT questionnaire seems to be a valid and useful instrument to evaluate the participants' understanding of the ICF. In this study, we found that some concepts, like 'study protocol' or 'randomization', were not well understood by participants when signing the ICF, especially by participants with lower education levels. They also believed that the experimental intervention would solve their health condition. Greater awareness about the importance of the informed consent process and ICF is necessary so that participants can fully understand the protocol, especially the risks involved, and their rights as participants.

Multiple karyotype differences between populations of the Hoplias malabaricus (Teleostei; Characiformes), a species complex in the gray area of the speciation process.

Neotropical fishes exhibit remarkable karyotype diversity, whose evolution is poorly understood. Here, we studied genetic differences in 60 individuals, from 11 localities of one species, the wolf fish Hoplias malabaricus, from populations that include six different "karyomorphs". These differ in Y-X chromosome differentiation, and, in several cases, by fusions with autosomes that have resulted in multiple sex chromosomes. Other differences are also observed in diploid chromosome numbers and morphologies. In an attempt to start understanding how this diversity was generated, we analyzed within- and between-population differences in a genome-wide sequence data set. We detect clear genotype differences between karyomorphs. Even in sympatry, samples with different karyomorphs differ more in sequence than samples from allopatric populations of the same karyomorph, suggesting that they represent populations that are to some degree reproductively isolated. However, sequence divergence between populations with different karyomorphs is remarkably low, suggesting that chromosome rearrangements may have evolved during a brief evolutionary time. We suggest that the karyotypic differences probably evolved in allopatry, in small populations that would have allowed rapid fixation of rearrangements, and that they became sympatric after their differentiation. Further studies are needed to test whether the karyotype differences contribute to reproductive isolation detected between some H. malabaricus karyomorphs.

The role of obstructive sleep apnea and nocturnal hypoxia as predictors of mortality in cancer patients.

BACKGROUND: Obstructive sleep apnea (OSA), due to its high prevalence, has been associated with a number of comorbidities, frequently impacting the overall course of these other diseases if left untreated. Recent studies highlight a potential association between OSA and cancer. This study investigates how OSA severity and hypoxia affect cancer prognosis, aiming to elucidate how they interplay. METHODS: Retrospective study including patients with a diagnosis of OSA after any cancer type followed up in a tertiary center during a 10-year period. OSA was mainly diagnosed after level III polysomnographic studies. RESULTS: Nocturnal hypoxia was significantly more prevalent in patients presenting lung cancer versus other malignancies and was associated with higher rates of oncologic disease progression. Overall survival was significantly lower in severe OSA patients and also in patients presenting nocturnal hypoxia. A composite hypoxia score considering both OSA severity and significant hypoxia was an independent predictor of mortality regardless of clinical cancer staging and treatment. Shorter time between cancer and OSA diagnosis was also associated with worse prognosis. CONCLUSION: This study suggests an association between OSA severity and nocturnal hypoxia and increased cancer mortality independently from possible confounding factors such as age, cancer clinical staging at diagnosis, treatment modality and also progression. Neoplastic patients with severe OSA and/or complex hypoxia seem to have lower overall survival rates than those with less severe OSA and nocturnal hypoxia.

Time since fire as a driver of taxonomic and phylogenetic patterns of grassland plant communities.

Fire plays a key role in grasslands, determining the distribution and evolution of species and boundaries with neighboring ecosystems. Evidence of community-wide responses to fire is largely based on taxonomic and functional descriptors, while the phylogenetic dimension is overlooked. Here we evaluated how the taxonomic and phylogenetic structure of grassland plant communities responded to a time since fire (TSF) gradient. We sampled 12 communities in Southern Brazil under varying TSF and calculated taxonomic species richness (S) and dominance (D), phylogenetic diversity (PD), and mean phylogenetic distances (MPD). We used Structural Equation Models to test the relationships between the environmental gradient and community descriptors. Communities with longer TSF presented higher PD and MPD but lower species richness and increased taxonomic dominance. These sites were dominated by monocots, specifically C4 grasses, but also presented exclusive clades, whereas recently-burned sites presented lower taxonomic dominance and more species distributed in a wider variety of clades. Our results indicate that these scenarios are interchangeable and dependent on fire management. Fire adaptation was not constrained by phylogenetic relatedness, contrasting with previous findings for tropical savannahs and indicating that temperate and tropical non-forest ecosystems from South America respond differently to fire, possibly due to different evolutionary histories.

Pulmonary Hypertension Induced by Right Pulmonary Artery Occlusion: Hemodynamic Consequences of Bmpr2 Mutation.

BACKGROUND: The primary genetic risk factor for heritable pulmonary arterial hypertension is the presence of monoallelic mutations in the BMPR2 gene. The incomplete penetrance of BMPR2 mutations implies that additional triggers are necessary for pulmonary arterial hypertension occurrence. Pulmonary artery stenosis directly raises pulmonary artery pressure, and the redirection of blood flow to unobstructed arteries leads to endothelial dysfunction and vascular remodeling. We hypothesized that right pulmonary artery occlusion (RPAO) triggers pulmonary hypertension (PH) in rats with Bmpr2 mutations. METHODS AND RESULTS: Male and female rats with a 71 bp monoallelic deletion in exon 1 of Bmpr2 and their wild-type siblings underwent acute and chronic RPAO. They were subjected to full high-fidelity hemodynamic characterization. We also examined how chronic RPAO can mimic the pulmonary gene expression pattern associated with installed PH in unobstructed territories. RPAO induced precapillary PH in male and female rats, both acutely and chronically. Bmpr2 mutant and male rats manifested more severe PH compared with their counterparts. Although wild-type rats adapted to RPAO, Bmpr2 mutant rats experienced heightened mortality. RPAO induced a decline in cardiac contractility index, particularly pronounced in male Bmpr2 rats. Chronic RPAO resulted in elevated pulmonary IL-6 (interleukin-6) expression and decreased Gdf2 expression (corrected P value<0.05 and log2 fold change>1). In this context, male rats expressed higher pulmonary levels of endothelin-1 and IL-6 than females. CONCLUSIONS: Our novel 2-hit rat model presents a promising avenue to explore the adaptation of the right ventricle and pulmonary vasculature to PH, shedding light on pertinent sex- and gene-related effects.

Association of adverse childhood experiences with dietary patterns of school-age children: evidence from the birth cohort Generation XXI.

BACKGROUND: Adverse Childhood Experiences (ACEs) have been associated with health-risk behaviors and several chronic diseases in adulthood. However, the relationship between exposure to ACEs and dietary patterns at school age is unknown. OBJECTIVES: To investigate the association between ACEs and dietary patterns of 10-year-olds. METHODS: The study included 5034 children from the Generation XXI cohort, recruited in 2005/2006 in Porto, Portugal. ACEs were assessed through a self-administered questionnaire covering the first 10 years (y) of life, quantified and grouped into 5 dimensions: "abuse," "school problems," "death/severe disease," "life changes," and "household dysfunction." Dietary patterns were identified by latent class analysis using data collected with a validated food frequency questionnaire. Five dietary patterns were studied: "low consumption," "energy-dense foods," "snacking," "intermediate consumption," and "healthier" (used as reference). Multinomial regression analyses were conducted, adjusted for the child's sex, household income, family structure, and mother's age [odds ratio (OR) and 99% confidence intervals (CIs)]. RESULTS: Most children were exposed to ≥1 ACE (96%), and ∼27% had reported 6 or more ACEs throughout life. Those reporting 4-5 and ≥6 ACEs were more likely to follow the "Energy-dense foods" dietary pattern compared with those with no ACEs (OR: 2.41; 99% CI: 1.00, 5.77 and OR: 2.65; 99% CI: 1.10, 6.39, respectively). Children exposed to "abuse" in the first 10 y showed 28% higher odds of following the "low consumption" dietary pattern when compared to children with no reported ACEs and using the "healthier" dietary pattern as a reference (OR: 1.28; 99% CI: 1.00, 1.63). CONCLUSIONS: Exposure to ACEs was associated with less healthy dietary patterns in school-aged children. Results suggest a cumulative effect of the adverse experiences resulting in a dietary pattern higher in energy-dense foods. Children with ACEs reported under the dimension of "abuse" seemed to have reduced food consumption.

The effects of field strength on stimulated echo and motion-compensated spin-echo diffusion tensor cardiovascular magnetic resonance sequences.

BACKGROUND: In-vivo diffusion tensor cardiovascular magnetic resonance (DT-CMR) is an emerging technique for microstructural tissue characterization in the myocardium. Most studies are performed at 3T, where higher signal-to-noise ratio (SNR) should benefit this signal-starved method. However, a few studies have suggested that DT-CMR is possible at 1.5T, where echo planar imaging artifacts may be less severe and 1.5T hardware is more widely available. METHODS: We recruited 20 healthy volunteers and performed mid-ventricular short-axis DT-CMR at 1.5T and 3T. Acquisitions were performed at peak systole and end-diastole using both stimulated echo acquisition mode (STEAM) and motion-compensated spin-echo (MCSE) sequences at matched spatial resolutions. DT-CMR parameters were averaged over the left ventricle and compared between 1.5T and 3T sequences using both datasets with and without the blow reference data included. RESULTS: Eleven (1.5T) and 12 (3T) diastolic MCSE acquisitions were rejected as the helix angle (HA) demonstrated <50% normal appearance circumferentially or the acquisition was abandoned due to poor image quality; a maximum of one acquisition was rejected for other datasets. Subjective HA map quality was significantly better at 3T than 1.5T for STEAM (p < 0.05), but not for MCSE and other DT-CMR quality measures were consistent with improvements in STEAM at 3T over 1.5T. When blow data were excluded, no significant differences in mean diffusivity were observed between field strengths, but fractional anisotropy was significantly higher at 1.5T than 3T for STEAM systole (p < 0.05). Absolute second eigenvector orientation (E2A, sheetlet angle) was significantly higher at 1.5T than 3T for MCSE systole and STEAM diastole, but significantly lower for STEAM systole (all p < 0.05). Transmural HA distribution was less steep at 1.5T than 3T for STEAM diastole data (p < 0.05). SNR was higher at 3T than 1.5T for all acquisitions (p < 0.05). CONCLUSION: While 3T provides benefits in terms of SNR, both STEAM and MCSE can be performed at 1.5T. However, MCSE is unreliable in diastole at both field strengths and STEAM benefits from the improved SNR at 3T over 1.5T. Future clinical research studies may be able to leverage the wider availability of 1.5T CMR hardware where MCSE acquisitions are desirable.

Revisiting the reaction pathways for phospholipid hydrolysis catalyzed by phospholipase A2 with QM/MM methods.

Secreted phospholipase A2 (sPLA2) is a Ca2+-dependent, widely distributed enzyme superfamily in almost all mammalian tissues and bacteria. It is also a critical component of the venom of nearly all snakes, as well as many invertebrate species. In non-venomous contexts, sPLA2 assumes significance in cellular signaling pathways by binding cell membranes and catalyzing ester bond hydrolysis at the sn-2 position of phospholipids. Elevated levels of GIIA sPLA2 have been detected in the synovial fluid of arthritis patients, where it exhibits a pro-inflammatory function. Consequently, identifying sPLA2 inhibitors holds promise for creating highly effective pharmaceutical treatments. Beyond arthritis, the similarities among GIIA sPLA2s offer an opportunity for developing treatments against snakebite envenoming, the deadliest neglected tropical disease. Despite decades of study, the details of PLA2 membrane-binding, substrate-binding, and reaction mechanism remain elusive, demanding a comprehensive understanding of the sPLA2 catalytic mechanism. This study explores two reaction mechanism hypotheses, involving one or two water molecules, and distinct roles for the Ca2+ cofactor. Our research focuses on the human synovial sPLA2 enzyme bound to lipid bilayers of varying phospholipid compositions, and employing adiabatic QM/MM and QM/MM MD umbrella sampling methods to energetically and geometrically characterize the structures found along both reaction pathways. Our studies demonstrate the higher frequency of productive conformations within the single-water pathway, also revealing a lower free energy barrier for hydrolyzing POPC. Furthermore, we observe that the TS of this concerted one-step reaction closely resembles transition state geometries observed in X-ray crystallography complexes featuring high-affinity transition state analogue inhibitors.