m6A modification inhibits miRNAs' intracellular function, favoring their extracellular export for intercellular communication.

Epitranscriptomics represents a further layer of gene expression regulation. Specifically, N6-methyladenosine (m6A) regulates RNA maturation, stability, degradation, and translation. Regarding microRNAs (miRNAs), while it has been reported that m6A impacts their biogenesis, the functional effects on mature miRNAs remain unclear. Here, we show that m6A modification on specific miRNAs weakens their coupling to AGO2, impairs their function on target mRNAs, determines their delivery into extracellular vesicles (EVs), and provides functional information to receiving cells. Mechanistically, the intracellular functional impairment is caused by m6A-mediated inhibition of AGO2/miRNA interaction, the EV loading is favored by m6A-mediated recognition by the RNA-binding protein (RBP) hnRNPA2B1, and the EV-miRNA function in the receiving cell requires their FTO-mediated demethylation. Consequently, cells express specific miRNAs that do not impact endogenous transcripts but provide regulatory information for cell-to-cell communication. This highlights that a further level of complexity should be considered when relating cellular dynamics to specific miRNAs.

m6a methylation orchestrates IMP1 regulation of microtubules during human neuronal differentiation.

Neuronal differentiation requires building a complex intracellular architecture, and therefore the coordinated regulation of defined sets of genes. RNA-binding proteins (RBPs) play a key role in this regulation. However, while their action on individual mRNAs has been explored in depth, the mechanisms used to coordinate gene expression programs shaping neuronal morphology are poorly understood. To address this, we studied how the paradigmatic RBP IMP1 (IGF2BP1), an essential developmental factor, selects and regulates its RNA targets during the human neuronal differentiation. We perform a combination of system-wide and molecular analyses, revealing that IMP1 developmentally transitions to and directly regulates the expression of mRNAs encoding essential regulators of the microtubule network, a key component of neuronal morphology. Furthermore, we show that m6A methylation drives the selection of specific IMP1 mRNA targets and their protein expression during the developmental transition from neural precursors to neurons, providing a molecular principle for the onset of target selectivity.

Semaglutide Effects on Metabolic Outcomes in Diabetes Mellitus Patients - Real World Study.

Purpose: Diabetes is a public health problem that requires strategies to impact glycemic control and reduce the risk of long-term medical complications. Pharmacological management is a necessary treatment for this disease. Therefore, semaglutide is an essential tool to achieve the treatment targets. The present study aimed to evaluate the semaglutide effects on a cohort with type 2 diabetes mellitus (T2DM) in Colombia. Materials and Methods: The cohort included 49 patients with T2DM that have been treated in a specialized care center. Their glycemic outcomes, weight, renal function, and adverse events were evaluated through a 3-, 6- and 12-month follow-up. Results: Significant differences were observed in the outcome evaluation: reduction of glycated hemoglobin levels (MD -2.74 CI -1.95 to -3.52 in 6 months), fasting plasma glucose levels, body weight (MD -7.11 CI -5.97 to -8.24), and the albumin-to-creatinine ratio. The results were maintained throughout the treatment period. The adverse event rate was 16.3%, predominating gastrointestinal events. Conclusion: This real-world evidence shows the efficacy of semaglutide in achieving treatment goals in patients with T2DM.

From the Operating Room to the Laboratory: Role of the Neuroscience Tissue Biorepository in the Clinical, Translational, and Basic Science Research Pipeline.

OBJECTIVE: To establish a neurologic disorder-driven biospecimen repository to bridge the operating room with the basic science laboratory and to generate a feedback cycle of increased institutional and national collaborations, federal funding, and human clinical trials. METHODS: Patients were prospectively enrolled from April 2017 to July 2022. Tissue, blood, cerebrospinal fluid, bone marrow aspirate, and adipose tissue were collected whenever surgically safe. Detailed clinical, imaging, and surgical information was collected. Neoplastic and nonneoplastic samples were categorized and diagnosed in accordance with current World Health Organization classifications and current standard practices for surgical pathology at the time of surgery. RESULTS: A total of 11,700 different specimens from 813 unique patients have been collected, with 14.2% and 8.5% of patients representing ethnic and racial minorities, respectively. These include samples from a total of 463 unique patients with a primary central nervous system tumor, 88 with metastasis to the central nervous system, and 262 with nonneoplastic diagnoses. Cerebrospinal fluid and adipose tissue dedicated banks with samples from 130 and 16 unique patients, respectively, have also been established. Translational efforts have led to 42 new active basic research projects; 4 completed and 6 active National Institutes of Health-funded projects; and 2 investigational new drug and 5 potential Food and Drug Administration-approved phase 0/1 human clinical trials, including 2 investigator initiated and 3 industry sponsored. CONCLUSION: We established a comprehensive biobank with detailed notation with broad potential that has helped us to transform our practice of research and patient care and allowed us to grow in research and clinical trials in addition to providing a source of tissue for new discoveries.

Development of a specific and potent IGF2BP1 inhibitor: A promising therapeutic agent for IGF2BP1-expressing cancers.

IGF2BP1 is a protein that controls the stability, localization, and translation of various mRNA targets. Poor clinical outcomes in numerous cancer types have been associated with its overexpression. As it has been demonstrated to impede tumor growth and metastasis in animal models, inhibiting IGF2BP1 function is a promising strategy for combating cancer. A lead chemical, 7773, which specifically decreased IGF2BP1 RNA binding and cellular activities, was previously identified in a high-throughput screen for effective IGF2BP1 inhibitors. Additional optimization of 7773 described in this manuscript led to the discovery of six compounds that performed equally well or better than 7773. In cell lines with high levels of endogenous IGF2BP1, one of 7773 derivatives, AVJ16, was found to be most efficient at preventing cell migration. Further, AVJ16 was found to be IGF2BP1-specific because it had no effect on cell lines that expressed little or no IGF2BP1 protein. The direct binding of AVJ16 to IGF2BP1 was validated by binding tests, with a 12-fold increase in binding efficiency over the lead compound. AVJ16 was shown to bind to a hydrophobic region at the protein's KH34 di-domain interface between the KH3 and KH4 domains. Overall, the findings imply that AVJ16 is a potent and specific inhibitor of IGF2BP1 activity.

Affinity-enhanced RNA-binding domains as tools to understand RNA recognition.

Understanding how the RNA-binding domains of a protein regulator are used to recognize its RNA targets is a key problem in RNA biology, but RNA-binding domains with very low affinity do not perform well in the methods currently available to characterize protein-RNA interactions. Here, we propose to use conservative mutations that enhance the affinity of RNA-binding domains to overcome this limitation. As a proof of principle, we have designed and validated an affinity-enhanced K-homology (KH) domain mutant of the fragile X syndrome protein FMRP, a key regulator of neuronal development, and used this mutant to determine the domain's sequence preference and to explain FMRP recognition of specific RNA motifs in the cell. Our results validate our concept and our nuclear magnetic resonance (NMR)-based workflow. While effective mutant design requires an understanding of the underlying principles of RNA recognition by the relevant domain type, we expect the method will be used effectively in many RNA-binding domains.

Direct m6A recognition by IMP1 underlays an alternative model of target selection for non-canonical methyl-readers.

m6A methylation provides an essential layer of regulation in organismal development, and is aberrant in a range of cancers and neuro-pathologies. The information encoded by m6A methylation is integrated into existing RNA regulatory networks by RNA binding proteins that recognise methylated sites, the m6A readers. m6A readers include a well-characterised class of dedicated proteins, the YTH proteins, as well as a broader group of multi-functional regulators where recognition of m6A is only partially understood. Molecular insight in this recognition is essential to build a mechanistic understanding of global m6A regulation. In this study, we show that the reader IMP1 recognises the m6A using a dedicated hydrophobic platform that assembles on the methyl moiety, creating a stable high-affinity interaction. This recognition is conserved across evolution and independent from the underlying sequence context but is layered upon the strong sequence specificity of IMP1 for GGAC RNA. This leads us to propose a concept for m6A regulation where methylation plays a context-dependent role in the recognition of selected IMP1 targets that is dependent on the cellular concentration of available IMP1, differing from that observed for the YTH proteins.

Offerings and User Demands of eHealth Services in Spain: National Survey.

BACKGROUND: The accelerated development of information and communication technologies has made health care one of the pioneering fields in the incorporation of these tools. As new technologies have been applied, existing technologies have been sophisticated and improved and the concept of eHealth has expanded. However, these advances and expansion of eHealth do not seem to have served to adapt the supply of services to users' demands; rather, supply seems to be governed by other variables. OBJECTIVE: The main objective of this work was to review the existing differences between user demands and the supply of eHealth services in Spain and their causes. The aim is to provide information on the level of use of the services and the causes of the variation in demand for these services, which can be useful in correcting existing differences and adapting them to the needs of users. METHODS: A survey, "Use and Attitudes Toward eHealth in Spain," was applied by telephone to a sample of 1695 people aged 18 years and over, taking into account sociodemographic profile characteristics (sex, age, habitat, educational level). The confidence level was set at 95% and the margin of error was ±2.45 for the whole sample. RESULTS: The survey results showed that the online doctor's appointment service is the most frequently used eHealth service by users: 72.48% of respondents used this service at some point and 21.28% stated that they use it regularly. The other services showed significantly lower percentages of use, including "managing health cards" (28.04%), "consulting medical history" (20.37%), "managing test results" (20.22%), "communicating with health professionals" (17.80%), and "requesting a change of doctor" (13.76%). Despite this low usage, a large majority of respondents (80.00%) attach great importance to all the services offered. Overall, 16.52% of the users surveyed were willing to make new service requests to the regional websites, with 9.33% of them highlighting services such as "the availability of a complaints and claims mailbox," "the possibility of consulting medical records," and "the availability of more detailed information on medical centers (location, medical directory, waiting lists, etc)." Other outstanding requests (8.00%) were to simplify the procedures for using certain existing services. CONCLUSIONS: The data from the survey show that eHealth services are widely known and highly valued by users, but not all services are used with the same frequency or intensity. It appears that users find it difficult to suggest new services that might be useful to them in terms of demand for new services that do not currently exist. It would be useful to use qualitative studies to gain a deeper understanding of currently unmet needs and the possibilities of eHealth. The lack of access to and use of these services and the unmet needs particularly affect more vulnerable populations who have the greatest difficulty in meeting their needs through alternative means to eHealth.

Physiological intron retaining transcripts in the cytoplasm abound during human motor neurogenesis.

Intron retention (IR) is now recognized as a dominant splicing event during motor neuron (MN) development; however, the role and regulation of intron-retaining transcripts (IRTs) localized to the cytoplasm remain particularly understudied. Here we show that IR is a physiological process that is spatiotemporally regulated during MN lineage restriction and that IRTs in the cytoplasm are detected in as many as 13% (n = 2297) of the genes expressed during this process. We identify a major class of cytoplasmic IRTs that are not associated with reduced expression of their own genes but instead show a high capacity for RNA-binding protein and miRNA occupancy. Finally, we show that ALS-causing VCP mutations lead to a selective increase in cytoplasmic abundance of this particular class of IRTs, which in turn temporally coincides with an increase in the nuclear expression level of predicted miRNA target genes. Altogether, our study identifies a previously unrecognized class of cytoplasmic intronic sequences with potential regulatory function beyond gene expression.

Tratamiento farmacológico del paciente que vive con diabetes mellitus tipo 2 / Pharmacological treatment of the patient living with type 2 diabetes mellitus

Resumen Introducción: el abordaje del paciente con diabetes mellitus tipo 2 ha cambiado a lo largo del tiempo. Años anteriores el enfoque terapéutico giraba en torno al nivel de hemoglobina glicada (HbA1c); ahora, con el advenimiento de nuevas moléculas, y luego de entender que la meta terapéutica no es sólo un punto de corte de HbA1c, se ha modificado el objetivo, para estar centrado en el paciente, en sus comorbilidades macro y microvasculares, riesgo cardiovascular y falla cardíaca. En esta revisión de la literatura se exponen las preguntas que se deben resolver al momento de definir una estrategia terapéutica en el paciente con diabetes mellitus tipo 2. Métodos: valoración de guías locales e internacionales de tratamiento de diabetes mellitus tipo 2. Resultados: se valoraron las guías del tratamiento de la diabetes mellitus tipo 2 realizadas por la Asociación Colombiana de Endocrinología, Asociación Colombiana de Medicina Interna, Asociación Colombiana de Nefrología, Sociedad Colombiana de Cardiología y Cirugía Cardiovascular, American Diabetes Association, American Association of Clinical Endocrinology, European Association for the Study of Diabetes y European Society of Cardiology. Además, se revisaron los ensayos clínicos que respaldan el uso de los diferentes grupos farmacológicos disponibles para el tratamiento de los pacientes con diabetes mellitus tipo 2. Conclusión: es menester del clínico realizar el abordaje terapéutico de los pacientes con diabetes mellitus tipo 2 de forma individualizada, centrado en el riesgo cardiovascular, comorbilidades asociadas, riesgo de hipoglucemia y costos de la intervención brindada.

Abstract Introduction: the approach to the patient with diabetes has changed over the past decade. Some years ago, the primary goal of treatment was to achieve a certain level of A1c as a marker of glucose control. Now, with new molecules in the market that can modify the disease and its complications the treatment is focused on the individual patient, his micro and macrovascular complications and the risk of cardiovascular, heart failure and renal complications. In this review of the literature, we expose some questions that need to be solved when approaching the treatment of a patient with type 2 diabetes. Methods: we reviewed local and international guidelines on the treatment of type 2 diabetes. Results: we reviewed guidelines in the treatment of diabetes, local guidelines; Asociación Colombiana de Endocrinología, Asociación Colombiana de Medicina Interna, Asociación Colombiana de Nefrología, Sociedad Colombiana de Cardiología y Cirugía Cardiovascular. International guidelines; American Diabetes Association, American Association of Clinical Endocrinology, European Association for the Study of Diabetes y European Society of Cardiology Additionally, we revised clinical trial that support the use of the different drugs that are recommended by the guidelines as first, second- or third-line treatment. Conclusion: as clinicians we are expected to give the patient an individualized approach, based on their own comorbidities, risk of cardiovascular and renal disease and risk of hypoglycemia guided by national and international guidelines and latest clinical trial that support our approach.

The TH1 cell lineage-determining transcription factor T-bet suppresses TH2 gene expression by redistributing GATA3 away from TH2 genes.

Lineage-determining transcription factors (LD-TFs) drive the differentiation of progenitor cells into a specific lineage. In CD4+ T cells, T-bet dictates differentiation of the TH1 lineage, whereas GATA3 drives differentiation of the alternative TH2 lineage. However, LD-TFs, including T-bet and GATA3, are frequently co-expressed but how this affects LD-TF function is not known. By expressing T-bet and GATA3 separately or together in mouse T cells, we show that T-bet sequesters GATA3 at its target sites, thereby removing GATA3 from TH2 genes. This redistribution of GATA3 is independent of GATA3 DNA binding activity and is instead mediated by the T-bet DNA binding domain, which interacts with the GATA3 DNA binding domain and changes GATA3's sequence binding preference. This mechanism allows T-bet to drive the TH1 gene expression program in the presence of GATA3. We propose that redistribution of one LD-TF by another may be a common mechanism that could explain how specific cell fate choices can be made even in the presence of other transcription factors driving alternative differentiation pathways.

The Present and Future of eHealth in Spain From a Health Management Perspective.

eHealth has grown significantly over the last decade. The aim of this study was to determine the level of use of information and communication technologies for health care in Spain and identify the main barriers to development. We used a qualitative study based on data obtained from 8 Spanish autonomous communities through semi-structured, in-depth interviews with key informants in eHealth management and planning. Programs present varying degrees of implementation. Services such as electronic prescriptions, digital medical records, and appointment requests via the Internet are advanced and widespread; others, such as digital imaging, are advanced but not fully deployed; and some, such as telecare programs, are experimental. The study also revealed diverse levels of interoperability and barriers to the expansion of these technologies, which can be classified into 4 fields: technological, organizational, human, and economic. eHealth might evolve more slowly in the coming years. Unless the payoff is clearly seen, major budget cuts in the current economic climate will prevent the implementation of new projects. Programs that help reduce health spending are more likely to be implemented, to the detriment of projects involving simple techniques or even clear health care improvements.

Small molecule inhibitor of Igf2bp1 represses Kras and a pro-oncogenic phenotype in cancer cells.

Igf2bp1 is an oncofetal RNA binding protein whose expression in numerous types of cancers is associated with upregulation of key pro-oncogenic RNAs, poor prognosis, and reduced survival. Importantly, Igf2bp1 synergizes with mutations in Kras to enhance signalling and oncogenic activity, suggesting that molecules inhibiting Igf2bp1 could have therapeutic potential. Here, we isolate a small molecule that interacts with a hydrophobic surface at the boundary of Igf2bp1 KH3 and KH4 domains, and inhibits binding to Kras RNA. In cells, the compound reduces the level of Kras and other Igf2bp1 mRNA targets, lowers Kras protein, and inhibits downstream signalling, wound healing, and growth in soft agar, all in the absence of any toxicity. This work presents an avenue for improving the prognosis of Igf2bp1-expressing tumours in lung, and potentially other, cancer(s).

The distinct RNA-interaction modes of a small ZnF domain underlay TUT4(7) diverse action in miRNA regulation.

TUT4 and the closely related TUT7 are non-templated poly(U) polymerases required at different stages of development, and their mis-regulation or mutation has been linked to important cancer pathologies. While TUT4(7) interaction with its pre-miRNA targets has been characterized in detail, the molecular bases of the broader target recognition process are unclear. Here, we examine RNA binding by the ZnF domains of the protein. We show that TUT4(7) ZnF2 contains two distinct RNA binding surfaces that are used in the interaction with different RNA nucleobases in different targets, i.e that this small domain encodes diversity in TUT4(7) selectivity and molecular function. Interestingly and unlike other well-characterized CCHC ZnFs, ZnF2 is not physically coupled to the flanking ZnF3 and acts independently in miRNA recognition, while the remaining CCHC ZnF of TUT4(7), ZnF1, has lost its intrinsic RNA binding capability. Together, our data suggest that the ZnFs of TUT4(7) are independent units for RNA and, possibly, protein-protein interactions that underlay the protein's functional flexibility and are likely to play an important role in building its interaction network.

Equity policies in health plans: accessibility and something more?

OBJECTIVE: To examine the approach adopted by the health plans of the autonomous communities of Spain, verifying the weight given to the concept of equity; to detect referenced communities or situations, as well as to distinguish the perspective of approaching it, from access, equity or equalization. METHODS: Qualitative study, of content analysis using Nvivo12, carried out in 2020 on health plans in force since 2019 in the different regions (autonomous communities) of Spain. Sixteen current regional health plans were compiled to establish base categories (equity, accessibility and equality) and determine associated terms using Nvivo12, from which a content analysis was performed. RESULTS: The concept of equity is not emphasized in the regional health plans and its relevance is surpassed by the concepts of accessibility and equality. The use of these three concepts is associated with various categories indicating circumstances, conditions or groups to which the plans give greater attention. CONCLUSIONS: The results obtained coincide with previous studies on the contents and orientation of health plans, revealing a discrete presence of the concept of equity in the approaches adopted, although this does not undermine the alignment of health policies with the visions emanating from transnational organizations. It is detected the existence of a group to which special attention is given from the accessibility approach, the population with functional diversity.

Biolayer Interferometry: Protein-RNA Interactions.

RNA-binding proteins often contain multiple RNA-binding domains connected by short flexible linkers. This domain arrangement allows the protein to bind the RNA with greater affinity and specificity than would be possible with individual domains and sometimes to remodel its structure. It is therefore important to understand how multiple modules interact with RNA because it is the modular nature of these proteins which specifies their biological function. This chapter is concerned with the use of biolayer interferometry to study protein-RNA interactions.