Maternal mRNAs with distinct 3' UTRs define the temporal pattern of Ccnb1 synthesis during mouse oocyte meiotic maturation.

The final stages of female gamete maturation occur in the virtual absence of transcription, with gene expression driven by a program of selective unmasking, translation, and degradation of maternal mRNAs. Here we demonstrate that the timing of Ccnb1 mRNA translation in mouse oocytes is dependent on the presence of transcripts with different 3' untranslated regions (UTRs). This 3' UTR heterogeneity directs distinct temporal patterns of translational activation or repression. Inclusion or exclusion of cis-acting elements is responsible for these divergent regulations. Our findings reveal an additional layer of translation control through alternative polyadenylation usage required to fine-tune the timing of meiosis progression.

Peripheral axonal ensheathment is regulated by RalA GTPase and the exocyst complex.

Axon ensheathment is fundamental for fast impulse conduction and the normal physiological functioning of the nervous system. Defects in axonal insulation lead to debilitating conditions, but, despite its importance, the molecular players responsible are poorly defined. Here, we identify RalA GTPase as a key player in axon ensheathment in Drosophila larval peripheral nerves. We demonstrate through genetic analysis that RalA action through the exocyst complex is required in wrapping glial cells to regulate their growth and development. We suggest that the RalA-exocyst pathway controls the targeting of secretory vesicles for membrane growth or for the secretion of a wrapping glia-derived factor that itself regulates growth. In summary, our findings provide a new molecular understanding of the process by which axons are ensheathed in vivo, a process that is crucial for normal neuronal function.

Neural networks for parameter estimation in microstructural MRI: Application to a diffusion-relaxation model of white matter.

Specific features of white matter microstructure can be investigated by using biophysical models to interpret relaxation-diffusion MRI brain data. Although more intricate models have the potential to reveal more details of the tissue, they also incur time-consuming parameter estimation that may converge to inaccurate solutions due to a prevalence of local minima in a degenerate fitting landscape. Machine-learning fitting algorithms have been proposed to accelerate the parameter estimation and increase the robustness of the attained estimates. So far, learning-based fitting approaches have been restricted to microstructural models with a reduced number of independent model parameters where dense sets of training data are easy to generate. Moreover, the degree to which machine learning can alleviate the degeneracy problem is poorly understood. For conventional least-squares solvers, it has been shown that degeneracy can be avoided by acquisition with optimized relaxation-diffusion-correlation protocols that include tensor-valued diffusion encoding. Whether machine-learning techniques can offset these acquisition requirements remains to be tested. In this work, we employ artificial neural networks to vastly accelerate the parameter estimation for a recently introduced relaxation-diffusion model of white matter microstructure. We also develop strategies for assessing the accuracy and sensitivity of function fitting networks and use those strategies to explore the impact of the acquisition protocol. The developed learning-based fitting pipelines were tested on relaxation-diffusion data acquired with optimal and sub-optimal acquisition protocols. Networks trained with an optimized protocol were observed to provide accurate parameter estimates within short computational times. Comparing neural networks and least-squares solvers, we found the performance of the former to be less affected by sub-optimal protocols; however, model fitting networks were still susceptible to degeneracy issues and their use could not fully replace a careful design of the acquisition protocol.

Computing and visualising intra-voxel orientation-specific relaxation-diffusion features in the human brain.

Diffusion MRI techniques are used widely to study the characteristics of the human brain connectome in vivo. However, to resolve and characterise white matter (WM) fibres in heterogeneous MRI voxels remains a challenging problem typically approached with signal models that rely on prior information and constraints. We have recently introduced a 5D relaxation-diffusion correlation framework wherein multidimensional diffusion encoding strategies are used to acquire data at multiple echo-times to increase the amount of information encoded into the signal and ease the constraints needed for signal inversion. Nonparametric Monte Carlo inversion of the resulting datasets yields 5D relaxation-diffusion distributions where contributions from different sub-voxel tissue environments are separated with minimal assumptions on their microscopic properties. Here, we build on the 5D correlation approach to derive fibre-specific metrics that can be mapped throughout the imaged brain volume. Distribution components ascribed to fibrous tissues are resolved, and subsequently mapped to a dense mesh of overlapping orientation bins to define a smooth orientation distribution function (ODF). Moreover, relaxation and diffusion measures are correlated to each independent ODF coordinate, thereby allowing the estimation of orientation-specific relaxation rates and diffusivities. The proposed method is tested on a healthy volunteer, where the estimated ODFs were observed to capture major WM tracts, resolve fibre crossings, and, more importantly, inform on the relaxation and diffusion features along with distinct fibre bundles. If combined with fibre-tracking algorithms, the methodology presented in this work has potential for increasing the depth of characterisation of microstructural properties along individual WM pathways.

Requirements for Animal Experiments: Problems and Challenges.

In vivo models remain a principle screening tool in the drug discovery pipeline. Here, the challenges associated with the need for animal experiments, as well as their impact on research, individual/societal, and economic contexts are discussed. A number of alternatives that, with further development, optimization, and investment, may replace animal experiments are also revised.

Combined diffusion-relaxometry microstructure imaging: Current status and future prospects.

Microstructure imaging seeks to noninvasively measure and map microscopic tissue features by pairing mathematical modeling with tailored MRI protocols. This article reviews an emerging paradigm that has the potential to provide a more detailed assessment of tissue microstructure-combined diffusion-relaxometry imaging. Combined diffusion-relaxometry acquisitions vary multiple MR contrast encodings-such as b-value, gradient direction, inversion time, and echo time-in a multidimensional acquisition space. When paired with suitable analysis techniques, this enables quantification of correlations and coupling between multiple MR parameters-such as diffusivity, T1 , T2 , and T2∗ . This opens the possibility of disentangling multiple tissue compartments (within voxels) that are indistinguishable with single-contrast scans, enabling a new generation of microstructural maps with improved biological sensitivity and specificity.

Accuracy and precision of statistical descriptors obtained from multidimensional diffusion signal inversion algorithms.

In biological tissues, typical MRI voxels comprise multiple microscopic environments, the local organization of which can be captured by microscopic diffusion tensors. The measured diffusion MRI signal can, therefore, be written as the multidimensional Laplace transform of an intravoxel diffusion tensor distribution (DTD). Tensor-valued diffusion encoding schemes have been designed to probe specific features of the DTD, and several algorithms have been introduced to invert such data and estimate statistical descriptors of the DTD, such as the mean diffusivity, the variance of isotropic diffusivities, and the mean squared diffusion anisotropy. However, the accuracy and precision of these estimations have not been assessed systematically and compared across methods. In this article, we perform and compare such estimations in silico for a one-dimensional Gamma fit, a generalized two-term cumulant approach, and two-dimensional and four-dimensional Monte-Carlo-based inversion techniques, using a clinically feasible tensor-valued acquisition scheme. In particular, we compare their performance at different signal-to-noise ratios (SNRs) for voxel contents varying in terms of the aforementioned statistical descriptors, orientational order, and fractions of isotropic and anisotropic components. We find that all inversion techniques share similar precision (except for a lower precision of the two-dimensional Monte Carlo inversion) but differ in terms of accuracy. While the Gamma fit exhibits infinite-SNR biases when the signal deviates strongly from monoexponentiality and is unaffected by orientational order, the generalized cumulant approach shows infinite-SNR biases when this deviation originates from the variance in isotropic diffusivities or from the low orientational order of anisotropic diffusion components. The two-dimensional Monte Carlo inversion shows remarkable accuracy in all systems studied, given that the acquisition scheme possesses enough directions to yield a rotationally invariant powder average. The four-dimensional Monte Carlo inversion presents no infinite-SNR bias, but suffers significantly from noise in the data, while preserving good contrast in most systems investigated.

Diffusion tensor distribution imaging of an in vivo mouse brain at ultrahigh magnetic field by spatiotemporal encoding.

Diffusion tensor distribution (DTD) imaging builds on principles from diffusion, solid-state and low-field NMR spectroscopies, to quantify the contents of heterogeneous voxels as nonparametric distributions, with tensor "size", "shape" and orientation having direct relations to corresponding microstructural properties of biological tissues. The approach requires the acquisition of multiple images as a function of the magnitude, shape and direction of the diffusion-encoding gradients, leading to long acquisition times unless fast image read-out techniques like EPI are employed. While in previous in vivo human brain studies performed at 3 T this proved a viable option, porting these measurements to very high magnetic fields and/or to heterogeneous organs induces B0 - and B1 -inhomogeneity artifacts that challenge the limits of EPI. To overcome such challenges, we demonstrate here that high spatial resolution DTD of mouse brain can be carried out at 15.2 T with a surface-cryoprobe, by relying on SPatiotemporal ENcoding (SPEN) imaging sequences. These new acquisition and data-processing protocols are demonstrated with measurements on in vivo mouse brain, and validated with synthetic phantoms designed to mimic the diffusion properties of white matter, gray matter and cerebrospinal fluid. While still in need of full extensions to 3D mappings and of scanning additional animals to extract more general physiological conclusions, this work represents another step towards the model-free, noninvasive in vivo characterization of tissue microstructure and heterogeneity in animal models, at ≈0.1 mm resolutions.

DAZL and CPEB1 regulate mRNA translation synergistically during oocyte maturation.

Meiotic progression requires exquisitely coordinated translation of maternal messenger (m)RNA that has accumulated during oocyte growth. A major regulator of this program is the cytoplasmic polyadenylation element binding protein 1 (CPEB1). However, the temporal pattern of translation at different meiotic stages indicates the function of additional RNA binding proteins (RBPs). Here, we report that deleted in azoospermia-like (DAZL) cooperates with CPEB1 to regulate maternal mRNA translation. Using a strategy that monitors ribosome loading onto endogenous mRNAs and a prototypic translation target, we show that ribosome loading is induced in a DAZL- and CPEB1-dependent manner, as the oocyte reenters meiosis. Depletion of the two RBPs from oocytes and mutagenesis of the 3' untranslated regions (UTRs) demonstrate that both RBPs interact with the Tex19.1 3' UTR and cooperate in translation activation of this mRNA. We observed a synergism between DAZL and cytoplasmic polyadenylation elements (CPEs) in the translation pattern of maternal mRNAs when using a genome-wide analysis. Mechanistically, the number of DAZL proteins loaded onto the mRNA and the characteristics of the CPE might define the degree of cooperation between the two RBPs in activating translation and meiotic progression.

Engineered Multifunctional Albumin-Decorated Porous Silicon Nanoparticles for FcRn Translocation of Insulin.

The last decade has seen remarkable advances in the development of drug delivery systems as alternative to parenteral injection-based delivery of insulin. Neonatal Fc receptor (FcRn)-mediated transcytosis has been recently proposed as a strategy to increase the transport of drugs across the intestinal epithelium. FcRn-targeted nanoparticles (NPs) could hijack the FcRn transcytotic pathway and cross the epithelial cell layer. In this study, a novel nanoparticulate system for insulin delivery based on porous silicon NPs is proposed. After surface conjugation with albumin and loading with insulin, the NPs are encapsulated into a pH-responsive polymeric particle by nanoprecipitation. The developed NP formulation shows controlled size and homogeneous size distribution. Transmission electron microscopy (TEM) images show successful encapsulation of the NPs into pH-sensitive polymeric particles. No insulin release is detected at acidic conditions, but a controlled release profile is observed at intestinal pH. Toxicity studies show high compatibility of the NPs with intestinal cells. In vitro insulin permeation across the intestinal epithelium shows approximately fivefold increase when insulin is loaded into FcRn-targeted NPs. Overall, these FcRn-targeted NPs offer a toolbox in the development of targeted therapies for oral delivery of insulin.

Two-Dimensional Correlation of Isotropic and Directional Diffusion Using NMR.

Diffusion nuclear magnetic resonance (NMR) is a powerful technique for studying porous media, but yields ambiguous results when the sample comprises multiple regions with different pore sizes, shapes, and orientations. Inspired by solid-state NMR techniques for correlating isotropic and anisotropic chemical shifts, we propose a diffusion NMR method to resolve said ambiguity. Numerical data inversion relies on sparse representation of the data in a basis of radial and axial diffusivities. Experiments are performed on a composite sample with a cell suspension and a liquid crystal.

Sexual Compulsivity Scale, Compulsive Sexual Behavior Inventory, and Hypersexual Disorder Screening Inventory: Translation, Adaptation, and Validation for Use in Brazil.

Epidemiological, behavioral, and clinical data on sexual compulsivity in Brazil are very limited. This study sought to adapt and validate the Sexual Compulsivity Scale (SCS), the 22-item version of the Compulsive Sexual Behavior Inventory (CSBI-22), and the Hypersexual Disorder Screening Inventory (HDSI) for use in Brazil. A total of 153 participants underwent psychiatric assessment and completed self-reported measures. The adaptation process of the instruments from English to Portuguese followed the guidelines of the International Society for Pharmacoeconomics and Outcomes Research. The reliability and validity of the HDSI criteria were evaluated and the construct validity of all measures was examined. For the SCS and HDSI, factor analysis revealed one factor for each measure. For the CSBI-22, four factors were retained although we only calculated the scores of two factors (control and violence). All scores had good internal consistency (alpha >.75), presented high temporal stability (>.76), discriminated between patients and controls, and presented strong (ρ > .81) correlations with the Sexual Addiction Screening Test (except for the violence domain = .40) and moderate correlations with the Impulsive Sensation Seeking domain of the Zuckerman Kuhlman Personality Questionnaire (ρ between .43 and .55). The sensitivity of the HDSI was 71.93 % and the specificity was 100 %. All measures showed very good psychometric properties. The SCS, the HDSI, and the control domain of the CSBI-22 seemed to measure theoretically similar constructs, as they were highly correlated (ρ > .85). The findings support the conceptualization of hypersexuality as a cluster of problematic symptoms that are highly consistent across a variety of measures.

Poly(A)-binding proteins are functionally distinct and have essential roles during vertebrate development.

Translational control of many mRNAs in developing metazoan embryos is achieved by alterations in their poly(A) tail length. A family of cytoplasmic poly(A)-binding proteins (PABPs) bind the poly(A) tail and can regulate mRNA translation and stability. However, despite the extensive biochemical characterization of one family member (PABP1), surprisingly little is known about their in vivo roles or functional relatedness. Because no information is available in vertebrates, we address their biological roles, establishing that each of the cytoplasmic PABPs conserved in Xenopus laevis [PABP1, embryonic PABP (ePABP), and PABP4] is essential for normal development. Morpholino-mediated knockdown of PABP1 or ePABP causes both anterior and posterior phenotypes and embryonic lethality. In contrast, depletion of PABP4 results mainly in anterior defects and lethality at later stages. Unexpectedly, cross-rescue experiments reveal that neither ePABP nor PABP4 can fully rescue PABP1 depletion, establishing that PABPs have distinct functions. Comparative analysis of the uncharacterized PABP4 with PABP1 and ePABP shows that it shares a mechanistically conserved core role in promoting global translation. Consistent with this analysis, each morphant displays protein synthesis defects, suggesting that their roles in mRNA-specific translational regulation and/or mRNA decay, rather than global translation, underlie the functional differences between PABPs. Domain-swap experiments reveal that the basis of the functional specificity is complex, involving multiple domains of PABPs, and is conferred, at least in part, by protein-protein interactions.

NMR molecular dynamics study of chromonic liquid crystals Edicol Sunset Yellow doped with salts.

We investigate the effect of monoatomic salts on the molecular dynamics in the nematic and isotropic phases formed by the chromonic liquid crystal Edicol Sunset Yellow. The study was carried out using proton nuclear magnetic resonance relaxometry. To analyse the effect of incorporation of additional sodium chloride or lithium chloride on the solutions' molecular dynamics, the spin-lattice relaxation time was measured for Larmor frequencies between 10 kHz and 100 MHz. In the nematic phase, the presence of additional sodium or lithium ions seems to contribute to an increase of the rotations/reorientations corr elation times in comparison with the mixture without extra ions. The collective motions detected by proton NMR relaxometry are associated with collective fluctuations of molecules within the stacks in the nematic phase and with order parameter fluctuations in the isotropic phase.

Health care expenses impact on the disability-adjusted life years in non-communicable diseases in the European Union.

Background: Non-communicable diseases are a global health problem. The metric Disability-Adjusted Life Years was developed to measure its impact on health systems. This metric makes it possible to understand a disease's burden, towards defining healthcare policies. This research analysed the effect of healthcare expenditures in the evolution of disability-adjusted life years for non-communicable diseases in the European Union between 2000 and 2019. Methods: Data were collected for all 27 European Union countries from Global Burden of Disease 2019, Global Health Expenditure, and EUROSTAT databases. Econometric panel data models were used to assess the impact of healthcare expenses on the disability-adjusted life years. Only models with a coefficient of determination equal to or higher than 10% were analysed. Results: There was a decrease in the non-communicable diseases with the highest disability-adjusted life years: cardiovascular diseases (-2,952 years/105 inhabitants) and neoplasms (-618 years/105 inhabitants). Health expenditure significantly decreased disability-adjusted life years for all analysed diseases (p < 0.01) unless for musculoskeletal disorders. Private health expenditure did not show a significant effect on neurological and musculoskeletal disorders (p > 0.05) whereas public health expenditure did not significantly influence skin and subcutaneous diseases (p > 0.05). Conclusion: Health expenditure have proved to be effective in the reduction of several diseases. However, some categories such as musculoskeletal and mental disorders must be a priority for health policies in the future since, despite their low mortality, they can present high morbidity and disability.

A meta-analysis on the role of sonication in the diagnosis of cardiac implantable electronic device-related infections.

Introduction: One of the biggest obstacles in diagnosing Implant-Associated Infections is the lack of infection criteria and standardized diagnostic methods. These infections present a wide range of symptoms, and their diagnosis can be hampered by the formation of microbial biofilms on the surface of implants. This study aimed to provide insight into the performance of sonication in the diagnosis of infections associated with Cardiac Implantable Electronic Devices, to help define a consensus on the algorithm for the microbial diagnosis of these infections. Methods: We carried out a systematic review with meta-analysis. The PRISMA methodology guidelines were followed, and an advanced search was carried out in PubMed and Web of Science, which enabled 8 articles to be included in the review, in which a meta-analysis was also carried out. QUADAS-2 was used to assess the risk of bias and effect measures were calculated to assess publication bias. Results: The overall sensitivity of the method was 0.823 (95% CI: 0.682-0.910) and the specificity was 0.632 (95% CI: 0.506-0.743). Discussion: These results suggest that sonication may offer advantages in diagnosing these infections. However, it is essential to approach these findings carefully and take into account the recommendations provided in the EHRA 2019 guidelines. This study highlights the importance of more effective diagnostic approaches for implantable medical device-associated infections to improve the quality of treatment and minimize the risks associated with these challenging medical conditions.

Poly[µ2-aqua-µ4-[1-(4-chloro-phen-yl)-4,4,4-tri-fluoro-butane-1,3-dionato]-potassium].

In the title compound, [K(C10H5ClO2F3)(H2O)] n , the two independent K(+) ions are located on a twofold rotation axis. For each of the cations, the distorted cubic coordination environment is defined by two F and four O atoms of symmetry-related 1,4-chloro-phenyl-4,4,4-tri-fluoro-butane-1,3-dionate anions and by two O atoms of water mol-ecules. The µ4-bridging character of the anion and the µ2-bridging of the water mol-ecule lead to the formation of layers parallel to (100). The coordinating water mol-ecules are also involved in O-H⋯O hydrogen bonds that reinforce the mol-ecular cohesion within the layers, which are stacked along [100]. The ß-diketonate anion is not planar, with an angle of 31.78 (10)° between the mean planes of the diketonate group and the chloro-phenyl ring.

4,7-Diphenyl-1,10-phenanthroline methanol hemisolvate.

The asymmetric unit of the title compound, C24H16N2·0.5CH3OH, is comprised of two independent bathophenanthroline mol-ecules (systematic name: 4,7-diphenyl-1,10-phenanthroline) and one methanol mol-ecule. The bathophenanthroline mol-ecules are not planar as there is a considerable rotation of all terminal phenyl rings with respect to the central phenanthroline units [dihedral angles in the range 52.21 (12)-62.14 (10)°]. In addition, a non-negligible torsion is apparent in one of the phenanthroline units: the angle between the mean planes of the two pyridine rings is 14.84 (13)°. The methanol solvent mol-ecule is linked to both N atoms of a bathophenanthroline mol-ecule through a bifurcated O-H⋯(N,N) hydrogen bond.

Evaluation of cell membrane-derived nanoparticles as therapeutic carriers for pancreatic ductal adenocarcinoma using an in vitro tumour stroma model.

Here, we fabricated nanoparticles made solely from the membrane of cells found in the pancreatic tumour's microenvironment (TME), like the human MiaPaCa-2 cells and M2-polarized macrophages. The cell membrane-derived nanoparticles (CMNPs) deriving from the MiaPaCa-2 cells (MPC2-CMNPs) were loaded with the chemotherapeutic drug paclitaxel (PTX), and the CMNPs deriving from M2-polarized macrophages (M2-CMNPs) were loaded with the colony-stimulating factor 1 receptor inhibitor, pexidartinib (PXDB). The CMNPs' thorough morphological and physicochemical characterisation was followed by an in-depth study of their targeting ability and the endocytosis pathway involved during their internalisation. An in vitro model of the desmoplastic stroma comprising cancer-associated fibroblast-mimicking cells and M2-polarized macrophages was also developed. The model was characterised by collagen and α-smooth muscle actin (α-SMA) expression (overexpressed in desmoplasia) and was used to assess the CMNPs' ability to cross the stroma and target the tumour cells. Moreover, we assessed the effect of PXDB-loaded M2-CMNPs on the expression of M1 (CD80/CD86) and M2 (CD206/CD209) polarisation markers on activated macrophages. Finally, we evaluated the PTX and PXDB-loaded CMNPs' effect on the viability of all the used TME cell lines alone or in combination. Overall, this pilot study showed the potential of the CMNPs to cross an in vitro stroma model and act synergistically to treat PDAC.

Demand for emergency services during the COVID-19 pandemic and disease burden: a case study in Portugal.

Background: The COVID-19 pandemic brought changes in the pattern of care use. A significant increase in the volume of emergencies was expected. However, a significant decrease was observed worldwide. Methods: An observational, analytical and cross-sectional study of all records of emergency episodes of patients aged 18 years or older admitted to the emergency services of the University of Porto Hospital Centre (2018-2022) were analysed. Results: During the pandemic, a significant reduction in emergency episode admissions (up to 40% during lockdowns), an increase in pre-emergency services, and discharges from Infectious Diseases and Internal Medicine was observed. The discharges from General Practice and General Practice and Family Medicine were residual. Conclusion: The lower use and type of use of emergency services during the COVID-19 pandemic had a negative impact on the disease burden. This could be prevented in future pandemics through the development of strategies to promote confidence in the use of health resources and establishing contingency plans for virtual assistance.