DELVE: feature selection for preserving biological trajectories in single-cell data.

Single-cell technologies can measure the expression of thousands of molecular features in individual cells undergoing dynamic biological processes. While examining cells along a computationally-ordered pseudotime trajectory can reveal how changes in gene or protein expression impact cell fate, identifying such dynamic features is challenging due to the inherent noise in single-cell data. Here, we present DELVE, an unsupervised feature selection method for identifying a representative subset of molecular features which robustly recapitulate cellular trajectories. In contrast to previous work, DELVE uses a bottom-up approach to mitigate the effects of confounding sources of variation, and instead models cell states from dynamic gene or protein modules based on core regulatory complexes. Using simulations, single-cell RNA sequencing, and iterative immunofluorescence imaging data in the context of cell cycle and cellular differentiation, we demonstrate how DELVE selects features that better define cell-types and cell-type transitions. DELVE is available as an open-source python package: https://github.com/jranek/delve .

Novel Approaches to Studying SLC13A5 Disease.

The role of the sodium citrate transporter (NaCT) SLC13A5 is multifaceted and context-dependent. While aberrant dysfunction leads to neonatal epilepsy, its therapeutic inhibition protects against metabolic disease. Notably, insights regarding the cellular and molecular mechanisms underlying these phenomena are limited due to the intricacy and complexity of the latent human physiology, which is poorly captured by existing animal models. This review explores innovative technologies aimed at bridging such a knowledge gap. First, I provide an overview of SLC13A5 variants in the context of human disease and the specific cell types where the expression of the transporter has been observed. Next, I discuss current technologies for generating patient-specific induced pluripotent stem cells (iPSCs) and their inherent advantages and limitations, followed by a summary of the methods for differentiating iPSCs into neurons, hepatocytes, and organoids. Finally, I explore the relevance of these cellular models as platforms for delving into the intricate molecular and cellular mechanisms underlying SLC13A5-related disorders.

Characterization of the MT-2 Treg-like cell line in the presence and absence of forkhead box P3 (FOXP3).

CD4+ forkhead box P3 (FOXP3)+ regulatory T cells (Tregs) are essential in maintaining immune tolerance and suppressing excessive immune responses. Tregs also contribute to tissue repair processes distinct from their roles in immune suppression. For these reasons, Tregs are candidates for targeted therapies for inflammatory and autoimmune diseases, and in diseases where tissue damage occurs. MT-2 cells, an immortalized Treg-like cell line, offer a model to study Treg biology and their therapeutic potential. In the present study, we use clustered regularly interspaced palindromic repeats (CRISPR)-mediated knockdown of FOXP3 in MT-2 cells to understand the transcriptional and functional changes that occur when FOXP3 is lost and to compare MT-2 cells with primary human Tregs. We demonstrate that loss of FOXP3 affects the transcriptome of MT-2 cells and that FOXP3's potential downstream targets include a wide range of transcripts that participate in the cell cycle, promote growth and contribute to inflammatory processes, but do not wholly simulate previously reported human primary Treg transcriptional changes in the absence of FOXP3. We also demonstrate that FOXP3 regulates cell cycling and proliferation, expression of molecules crucial to Treg function and MT-2 cell-suppressive activities. Thus, MT-2 cells offer opportunities to address regulatory T-cell functions in vitro.

Organotypic 3D Cell-Architecture Impacts the Expression Pattern of miRNAs-mRNAs Network in Breast Cancer SKBR3 Cells.

BACKGROUND: Currently, most of the research on breast cancer has been carried out in conventional two-dimensional (2D) cell cultures due to its practical benefits, however, the three-dimensional (3D) cell culture is becoming the model of choice in cancer research because it allows cell-cell and cell-extracellular matrix (ECM) interactions, mimicking the native microenvironment of tumors in vivo. METHODS: In this work, we evaluated the effect of 3D cell organization on the expression pattern of miRNAs (by Small-RNAseq) and mRNAs (by microarrays) in the breast cancer SKBR3 cell line and analyzed the biological processes and signaling pathways regulated by the differentially expressed protein-coding genes (DE-mRNAs) and miRNAs (DE-microRNAs) found in the organoids. RESULTS: We obtained well-defined cell-aggregated organoids with a grape cluster-like morphology with a size up to 9.2 × 105 µm3. The transcriptomic assays showed that cell growth in organoids significantly affected (all p < 0.01) the gene expression patterns of both miRNAs, and mRNAs, finding 20 upregulated and 19 downregulated DE-microRNAs, as well as 49 upregulated and 123 downregulated DE-mRNAs. In silico analysis showed that a subset of 11 upregulated DE-microRNAs target 70 downregulated DE-mRNAs. These genes are involved in 150 gene ontology (GO) biological processes such as regulation of cell morphogenesis, regulation of cell shape, regulation of canonical Wnt signaling pathway, morphogenesis of epithelium, regulation of cytoskeleton organization, as well as in the MAPK and AGE-RAGE signaling KEGG-pathways. Interestingly, hsa-mir-122-5p (Fold Change (FC) = 15.4), hsa-mir-369-3p (FC = 11.4), and hsa-mir-10b-5p (FC = 20.1) regulated up to 81% of the 70 downregulated DE-mRNAs. CONCLUSION: The organotypic 3D cell-organization architecture of breast cancer SKBR3 cells impacts the expression pattern of the miRNAs-mRNAs network mainly through overexpression of hsa-mir-122-5p, hsa-mir-369-3p, and hsa-mir-10b-5p. All these findings suggest that the interaction between cell-cell and cell-ECM as well as the change in the culture architecture impacts gene expression, and, therefore, support the pertinence of migrating breast cancer research from conventional cultures to 3D models.

RNA-binding deficient TDP-43 drives cognitive decline in a mouse model of TDP-43 proteinopathy.

TDP-43 proteinopathies including frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) are neurodegenerative disorders characterized by aggregation and mislocalization of the nucleic acid-binding protein TDP-43 and subsequent neuronal dysfunction. Here, we developed endogenous models of sporadic TDP-43 proteinopathy based on the principle that disease-associated TDP-43 acetylation at lysine 145 (K145) alters TDP-43 conformation, impairs RNA-binding capacity, and induces downstream mis-regulation of target genes. Expression of acetylation-mimic TDP-43K145Q resulted in stress-induced nuclear TDP-43 foci and loss of TDP-43 function in primary mouse and human-induced pluripotent stem cell (hiPSC)-derived cortical neurons. Mice harboring the TDP-43K145Q mutation recapitulated key hallmarks of FTLD, including progressive TDP-43 phosphorylation and insolubility, TDP-43 mis-localization, transcriptomic and splicing alterations, and cognitive dysfunction. Our study supports a model in which TDP-43 acetylation drives neuronal dysfunction and cognitive decline through aberrant splicing and transcription of critical genes that regulate synaptic plasticity and stress response signaling. The neurodegenerative cascade initiated by TDP-43 acetylation recapitulates many aspects of human FTLD and provides a new paradigm to further interrogate TDP-43 proteinopathies.

SOX9 knockout decreases stemness properties in colorectal cancer cells.

Background: Colorectal cancer (CRC) is a leading cause of death worldwide. SRY-box transcription factor 9 (SOX9) participates in organogenesis and cell differentiation in normal tissues but has been involved in carcinogenesis development. Cancer stem cells (CSCs) are a small population of cells present in solid tumors that contribute to increased tumor heterogeneity, metastasis, chemoresistance, and relapse. CSCs have properties such as self-renewal and differentiation, which can be modulated by many factors. Currently, the role of SOX9 in the maintenance of the stem phenotype has not been well elucidated, thus, in this work we evaluated the effect of the absence of SOX9 in the stem phenotype of CRC cells. Methods: We knockout (KO) SOX9 in the undifferentiated CRC cell line HCT116 and evaluated their stemness properties using sphere formation assay, differentiation assay, and immunophenotyping. Results: SOX9-KO affected the epithelial morphology of HCT116 cells and stemness characteristics such as its pluripotency signature with the increase of SOX2 as a compensatory mechanism to induce SOX9 expression, the increase of KLF4 as a differentiation feature, as well as the inhibition of the stem cell markers CD44 and CD73. In addition, SOX9-KO cells gain the epithelial-mesenchymal transition (EMT) phenotype with a significant upregulation of CDH2. Furthermore, our results showed a remarkable effect on first- and second-sphere formation, being SOX9-KO cells less capable of forming high-size-resistant spheres. Nevertheless, CSCs surface markers were not affected during the differentiation assay. Conclusions: Collectively, our findings supply evidence that SOX9 promotes the maintenance of stemness properties in CRC-CSCs.

Intermediate filament dysregulation in astrocytes in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN).

Giant Axonal Neuropathy (GAN) is a pediatric neurodegenerative disease caused by KLHL16 mutations. KLHL16 encodes gigaxonin, which regulates intermediate filament (IF) turnover. Previous neuropathological studies and examination of postmortem brain tissue in the current study revealed involvement of astrocytes in GAN. To develop a clinically-relevant model, we reprogrammed skin fibroblasts from seven GAN patients to pluripotent stem cells (iPSCs), which were used to generate neural progenitor cells (NPCs), astrocytes, and brain organoids. Multiple isogenic control clones were derived via CRISPR/Cas9 gene editing of one patient line carrying the G332R gigaxonin mutation. All GAN iPSCs were deficient for gigaxonin and displayed patient-specific increased vimentin expression. GAN NPCs had lower nestin expression and fewer nestin-positive cells compared to isogenic controls, but nestin morphology was unaffected. GAN brain organoids were marked by the presence of neurofilament and GFAP aggregates. GAN iPSC-astrocytes displayed striking dense perinuclear vimentin and GFAP accumulations and abnormal nuclear morphology. In over-expression systems, GFAP oligomerization and perinuclear aggregation were augmented in the presence of vimentin. GAN patient cells with large perinuclear vimentin aggregates accumulated significantly more nuclear KLHL16 mRNA compared to cells without vimentin aggregates. As an early effector of KLHL16 mutations, vimentin may be a potential target in GAN.

Modulation of tau tubulin kinases (TTBK1 and TTBK2) impacts ciliogenesis.

Tau tubulin kinase 1 and 2 (TTBK1/2) are highly homologous kinases that are expressed and mediate disease-relevant pathways predominantly in the brain. Distinct roles for TTBK1 and TTBK2 have been delineated. While efforts have been devoted to characterizing the impact of TTBK1 inhibition in diseases like Alzheimer's disease and amyotrophic lateral sclerosis, TTBK2 inhibition has been less explored. TTBK2 serves a critical function during cilia assembly. Given the biological importance of these kinases, we designed a targeted library from which we identified several chemical tools that engage TTBK1 and TTBK2 in cells and inhibit their downstream signaling. Indolyl pyrimidinamine 10 significantly reduced the expression of primary cilia on the surface of human induced pluripotent stem cells (iPSCs). Furthermore, analog 10 phenocopies TTBK2 knockout in iPSCs, confirming a role for TTBK2 in ciliogenesis.

Intermediate filament dysregulation and astrocytopathy in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN).

Giant Axonal Neuropathy (GAN) is a pediatric neurodegenerative disease caused by KLHL16 mutations. KLHL16 encodes gigaxonin, a regulator of intermediate filament (IF) protein turnover. Previous neuropathological studies and our own examination of postmortem GAN brain tissue in the current study revealed astrocyte involvement in GAN. To study the underlying mechanisms, we reprogrammed skin fibroblasts from seven GAN patients carrying different KLHL16 mutations to iPSCs. Isogenic controls with restored IF phenotypes were derived via CRISPR/Cas9 editing of one patient carrying a homozygous missense mutation (G332R). Neural progenitor cells (NPCs), astrocytes, and brain organoids were generated through directed differentiation. All GAN iPSC lines were deficient for gigaxonin, which was restored in the isogenic control. GAN iPSCs displayed patient-specific increased vimentin expression, while GAN NPCs had decreased nestin expression compared to isogenic control. The most striking phenotypes were observed in GAN iPSC-astrocytes and brain organoids, which exhibited dense perinuclear IF accumulations and abnormal nuclear morphology. GAN patient cells with large perinuclear vimentin aggregates accumulated nuclear KLHL16 mRNA. In over-expression studies, GFAP oligomerization and perinuclear aggregation were potentiated in the presence of vimentin. As an early effector of KLHL16 mutations, vimentin may serve as a potential therapeutic target in GAN.

Using liver models generated from human-induced pluripotent stem cells (iPSCs) for evaluating chemical-induced modifications and disease across liver developmental stages.

The liver is a pivotal organ regulating critical developmental stages of fetal metabolism and detoxification. Though numerous studies have evaluated links between prenatal/perinatal exposures and adverse health outcomes in the developing fetus, the central role of liver to health disruptions resulting from these exposures remains understudied, especially concerning early development and later-in-life health outcomes. While numerous in vitro methods for evaluating liver toxicity have been established, the use of iPSC-derived hepatocytes appears to be particularly well suited to contribute to this critical research gap due to their potential to model a diverse range of disease phenotypes and different stages of liver development. The following key aspects are reviewed: (1) an introduction to developmental liver toxicity; (2) an introduction to embryonic and induced pluripotent stem cell models; (3) methods and challenges for deriving liver cells from stem cells; and (4) applications for iPSC-derived hepatocytes to evaluate liver developmental stages and their associated responses to insults. We conclude that iPSC-derived hepatocytes have great potential for informing liver toxicity and underlying disease mechanisms via the generation of patient-specific iPSCs; implementing large-scale drug and chemical screening; evaluating general biological responses as a potential surrogate target cell; and evaluating inter-individual disease susceptibility and response variability.

Teaching Rheumatology in the undergraduate: It is enough? / Educación y reumatología en el pregrado: ¿enseñamos suficiente?

ABSTRACT Introduction: Musculoskeletal disorders (MSD) are the second leading cause of disability worldwide. There are difficulties in the early diagnosis and therapeutic approach to these pathologies, with a negative impact on their outcomes. Access to rheumatology is limited, with a low supply in the face of growing demand, which makes the general practitioner the first contact for care. Objectives: Describe the perception and confidence that general practitioners have regarding the training in rheumatology received at undergraduate level. Materials and methods: Observational cross-sectional study, with a Likert-type survey tool being used. The study included general practitioners graduated from the Colombian Medicine program between 2009 and 2019. The variables studied were those related to the curriculum, acquired knowledge or skills, and proficiency in content in rheumatol ogy compared to practice. Subjects who attended a specialist or who had an employment relationship with a specialist rheumatology centre were excluded. Results and Conclusions: A total of 102 physicians were surveyed, and 86 completed ques tionnaires were included in the final analysis. Of these, 83.4% were graduates of private universities. Over two-thirds (37%9) had a formal subject in rheumatology, 16% received training with specific strategies, 54% expressed security when performing the MS physical examination, and 47% were sure in the diagnostic approach, and prescription of disease-modifying drugs. In order to strengthen the training in rheumatology required by the undergraduate, a joint effort is required with the medical schools in defining the competen cies and skills of the primary care physician, together with the health needs and available educational strategies.

RESUMEN Introducción: Las enfermedades musculoesqueléticas (ME) son la segunda causa de discapa cidad mundial. Se presentan dificultades en el enfoque diagnóstico y terapéutico temprano de estas enfermedades, lo cual tiene un impacto negativo en sus desenlaces. El acceso a Reumatología es limitado, con una baja oferta frente a la creciente demanda, lo que convierte al médico general en el primer contacto de atención. Objetivos: Describir la percepción y la confianza que tienen los médicos generales respecto a la formación en reumatología recibida en el pregrado. Materiales y métodos: Estudio observacional de corte transversal en el cual se indagó a médi cos generales, egresados de programas de medicina colombianos entre el 2009 y 2019, mediante un cuestionario con respuesta tipo Likert, sobre variables relacionadas con el planteamiento curricular, los conocimientos o habilidades adquiridas y la suficiencia en el contenido en reumatología con respecto a la práctica. Se excluyó a los sujetos que cur saran algún programa de especialización o que tuvieran relación laboral con un centro especializado de Reumatología. Resultados y conclusiones: Se encuestó a 102 médicos; 86 encuestas fueron incluidas en el análisis final. El 83,4% de ellos eran egresados de universidades privadas, el 37% contó con una asignatura formal de reumatología, el 16% recibió formación con estrategias específicas, el 54% manifestó seguridad al realizar el examen físico musculoesquelético, el 47% expresó sentirse seguro en el enfoque diagnóstico y la prescripción de medicamentos modificadores de la enfermedad. Es necesario fortalecer la formación en Reumatología en el pregrado; se requiere un trabajo conjunto con las facultades de medicina en la definición de competen cias del médico de Atención Primaria, alineado con las necesidades de salud y las estrategias educacionales disponibles.

Mesoscience in cell biology and cancer research.

Mesoscale characteristics and their interdimensional correlation are the focus of contemporary interdisciplinary research. Mesoscience is a discipline that has the potential to radically update the existing knowledge structure, which differs from the conventional unit-scale and system-scale research models, revealing a previously untouchable area for scientific research. Integrative biology research aims to dissect the complex problems of life systems by conducting comprehensive research and integrating various disciplines from all biological levels of the living organism. However, the mesoscientific issues between different research units are neglected and challenging. Mesoscale research in biology requires the integration of research theories and methods from other disciplines (mathematics, physics, engineering, and even visual imaging) to investigate theoretical and frontier questions of biological processes through experiments, computations, and modeling. We reviewed  integrative paradigms and methods for the biological mesoscale problems (focusing on oncology research) and prospected the potential of their multiple dimensions and upcoming challenges. We expect to establish an interactive and collaborative theoretical platform for further expanding the depth and width of our understanding on the nature of biology.

Prevalencia de tuberculosis latente determinada mediante la prueba de derivado proteico purificado (PPD) en una población de pacientes adultos con artritis reumatoide llevados a terapia biotecnológica / Prevalence of latent tuberculosis determined using the Purified Protein Derivative (PPD) test in adult patients with rheumatoid arthritis receiving biotechnological therapy

RESUMEN Introducción: Colombia es un país endémico para tuberculosis (TB), con una prevalencia de 26 casos por millón. Sin embargo, no se cuenta con datos recientes y claros respecto a la prevalencia de tuberculosis latente (TBL) en la población con artritis reumatoide (AR) candidata a terapia biotecnológica. Metodología: Estudio de corte transversal con componente analítico para determinar la pre-valencia de TBL en pacientes con AR candidatos a terapia biotecnológica. Resultados: La prevalencia de TBL en pacientes con AR candidatos a terapia biotecnológica es alta, del 18,3% (IC 95% 14,7-21,9), y en los cruces exploratorios se encontró una relación entre TBL y la variable género masculino (p ≤ 0,001), hallazgos anormales en la radiografía de tórax (p = 0,039) y el tabaquismo (p = 0,028). Conclusión: La prevalencia de TBL en pacientes con AR candidatos a terapia biotecnológica es alta. Se requieren estudios prospectivos para evaluar la incidencia de TB en este grupo de pacientes y así corroborar su asociación.

ABSTRACT Introduction: Although tuberculosis (TB) is endemic in Columbia, with a prevalence of 26 cases per million, there are no recent and clear data regarding the prevalence of latent tuberculosis (LBT) in the population with rheumatoid arthritis (RA), candidates for biotechnological therapy. Methodology: A cross-sectional study with an analytical component to determine the prevalence of LBT in patients with RA who are candidates for biotechnological therapy. Results: The prevalence of LTB in RA patients who are candidates for biotechnological therapy is high, 18.3% (95% CI: 14.7-21.9). In the exploratory analysis, a relationship between LBT and male gender was found (P ≤ .001), as well as abnormal findings on chest radiography (P = .039), and smoking (P = .028). Conclusion: The prevalence of LTB in patients with RA who are candidates for biotechnological therapy is high. Prospective studies are needed to evaluate the incidence of TB in this group of patients and corroborate this association.

Generation of an induced pluripotent stem cell line (UNCCi002-A) from a healthy donor using a non-integration system to study Cerebral Cavernous Malformation (CCM).

The generation of induced pluripotent stem cells (iPSCs) from healthy individuals is an invaluable resource as reference control in disease modeling and drug discovery. This paper details the reprogramming of peripheral blood mononuclear cells (PBMCs) isolated from a healthy 27 years-old male using non-integration technology. The derived iPSCs displayed typical pluripotent stem cell morphology, the capacity to differentiate into the three germ layers, and normal karyotype. This iPSC line will be used as a reference control to study the Cerebral Cavernous Malformation disease mechanism.

Adherencia a medicamentos orales en pacientes con artritis reumatoide, una experiencia colombiana / Adherence to oral disease-modifying anti-rheumatic drugs in patients with rheumatoid arthritis: a Colombian experience

RESUMEN Introducción: El uso de los fármacos modificadores de la enfermedad (FAME) en pacientes con artritis reumatoide (AR) es esencial para alcanzar y mantener un control adecuado de la enfermedad y prevenir un daño funcional irreversible. Sin embargo, la tasa de adherencia a la terapia farmacológica varía entre el 20% y el 107%. Esta variabilidad puede deberse a los métodos de medición utilizados en los diferentes estudios. Objetivo: Evaluar la adherencia global al tratamiento oral con FAME en pacientes con AR mediante el autodiligenciamiento del cuestionario Compliance Questionnaire on Rheumatology (CQR) y la actividad de la enfermedad e identificar los factores potenciales asociados con la baja adherencia. Métodos: Estudio descriptivo transversal que incluyó pacientes mayores de 18 años con AR clasificados por criterios ACR-EULAR 2010, con prescripción de FAME durante al menos 3 meses y control con reumatólogo en el último año. Los participantes llenaron el cuestionario CQR y se les indagó acerca del conocimiento de la enfermedad, la percepción sobre el tratamiento, los efectos adversos, el índice de comorbilidad de Charlson, el índice global de apoyo social, el número de medicamentos y FAME prescritos, el acceso a los servicios de salud y la actividad de la enfermedad por DAS 28 o CDAI. Se definió como buena adherencia un punto de corte de CQR >80 y ausencia de actividad (remisión o actividad baja). Para la búsqueda de factores asociados con adherencia se analizaron los datos por medio de mediana y rango intercuartílico, así como frecuencias y proporciones. La comparación entre los grupos de adherentes y no adherentes se hizo con comparaciones absolutas, por medio de test de Mann-Whitney para las variables continuas y chi-cuadrado (para valores esperados > 5), o Fisher (para valor esperado < 5) para variables categóricas, tomando como nivel de significancia un valor de p<0,05. Se utilizaron OR y sus respectivos intervalos de confianza al 95% (IC95%). Resultados: De los 170 participantes incluidos, el 50% (n = 85) tuvo un valor de CQR mayor de 80 (buena adherencia). La mayoría de los pacientes se encontraba en remisión (60,6%) o baja actividad de la enfermedad (17%). El análisis posterior únicamente encontró asociación estadísticamente significativa entre adherencia medida por CQR y el número de amigos (p = 0,0012) y entre adherencia medida por actividad de la enfermedad y el índice de soporte social global (p = 0,004). Conclusiones: Este estudio muestra un nivel de adherencia similar al reportado en otras poblaciones, lo cual puede deberse a comportamientos propios de nuestra población, aunque los autores percibieron dificultades reportadas por los pacientes para entender los enunciados del instrumento en todos los niveles de escolaridad. Únicamente las variables de soporte social tuvieron una asociación estadísticamente significativa con la adherencia, asociación descrita en la literatura. Se requieren más estudios para evaluar las características operacionales del CQR en nuestra población.

ABSTRACT Introduction: The use of disease modifying antirheumatic drugs (DMARDs) in patients with rheumatoid arthritis (RA) is essential in order to achieve and maintain adequate disease control, and thus preventing irreversible functional damage. However, the rate of adherence to drug therapy has been reported to be between 20% and 107%. This variability may be due to the measurement methods used in the different studies. Objective: To test the overall medication adherence to oral treatment with DMARD in patients with RA using the self-report Spanish version Compliance Questionnaire on Rheumatology (CQR) and to identify potential factors associated with non-adherence. Methods: A cross-sectional descriptive study was conducted that included patients older than 18 years with RA diagnosed according to the ACR-EULAR 2010 criteria. They also had to have been prescribed oral DMARD for the previous 3 months, and had been seen by a rheumatologist in the last year. Patients completed the CQR to assess adherence, and were asked about disease knowledge, perception about treatment, side effects, Charlson Comorbidity Index, global index of social support, number of medications and DMARD prescribed, access to health resources, and disease activity measured by DAS 28 or CDAI. Good adherence was defined as a cut-off point of CQR > 80 or non-activity (remission or low activity). In the search for factors associated with adherence, the data were analysed using means of median and interquartile range, as well as frequencies and proportions. The comparison between adherent and non-adherent groups was performed using absolute comparisons, with the Mann-Whitney test for continuous, and chi-squared (for expected values > 5), or Fisher (for expected values < 5) tests for categorical variables, taking as a level of significance a value of P < .05. OR and their respective 95% confidence intervals (95% CI) were used. Results: Of 170 participants included, 50% (n = 85) had a value greater than 80% (good drug adherence). Most patients had remission (60.6%) or low disease activity (17%). The subsequent analysis showed statistically significant association between adherence measured by CQR and the number of friends (P = .0012). An association was also found between disease activity as an indirect indicator of adherence and the global social support index (P = .004). Conclusions: This study found a similar level of adherence to that reported in other populations, which could be due to the behaviour of our population, although the authors perceived difficulties reported by patients in understanding the statements of the questionnaire at all levels of education. Only the social support variables had a statistically significant relationship with adherence, which had also been described in the literature. Further studies are required to evaluate the operational characteristics of the CQR in our population.

P060 Screening Criteria of Inflammatory Bowel Disease: Application in Colombian Patients With Spondyloarthritis.

BACKGROUND: The Spondyloarthritis (SpA) is a group of chronic inflammatory rheumatic diseases, in which 5-10% of extra-articular manifestations are gastrointestinal such as the inflammatory bowel disease. Objective: To apply the clinical criteria for the screening of inflammatory bowel disease (IBD) in patients with SpA with gastrointestinal symptoms and its association with disease activity and function. METHODS: A Cross-sectional study included 82 patients with SpA, according to ASAS classification criteria without diagnosis of IBD. We applied the Screening criteria for IBD developed by Sanz et al, in the SpA patients. Clinical evaluation by rheumatologist and in patients with ≥ 2 gastrointestinal symptoms clinical evaluation by gastroenterologist and IBD screening criteria were performed. Digital chromoendoscopy, magnification colonoscopy, and histological analysis were performed. Lab tests included, C-reactive protein, sedimentation rate, serum levels of transferrin, ferritin and vitamin B12. The association between clinical variables and colonoscopy and histological variables were evaluated using the Chi-square or Fisher's exact test (Ethical / Cod. 2017-023). RESULTS: Of the 82 individuals evaluated, 58 of them were referred to gastroenterology with a direction to perform colonoscopy with chromeondospia, and 41 of them were able to intervene to whom the IBD screening criteria were applied. 53.7% are men, 7.3% actively smoke. 100% of the population presented some gastrointestinal symptoms, the most frequent being diarrhea of more than 4 weeks in 61%. 68.3% had at least one of the three major criteria. Rectorrhagia was associated with BASFI>4, p=0.050, axial compromise p = 0.043, diagnosis of PsA p = 0.090 and alterations in the architecture of the ileum p=0.034. Diarrhea was associated with ESR> 20, p = 0.050, BASFI>4 p = 0.012. In addition, 70.75 of the patients had at least one of the minor screening criteria associated with higher BASFI levels, p = 0.01. Aphthous stomatitis was reported as extra-intestinal manifestations in 7.3% and abdominal pain in 87.8% of the patients, which was associated with BASDAI>4 p = 0.023, ASDASCRP> 2.1, p = 0.043 and inflammation in the ileum, p = 0.046. No patients with positive iron deficiency anemia were found. However, ferritin alteration was observed in 22% associated with chronic inflammation of the colon, p = 0.042. There were no cases of fever or family history of IBD. Noting that in 17.1% of the cases a decrease in vitamin B12 levels was detected, associated with the presence of ulcers (p = 0.035) and acute inflammation in the ileum, p = 0.032. Weight loss was found in 31.7% of the cases and was associated with smoking history p = 0.039. CONCLUSION: We found a high frequency of major and minor symptoms of IBD, both of which were associated with a high activity of spondyloarthritis and an important functional compromise as well as inflammation markers in this group of patients. The application of the screening criteria for IBD in SpA without IBD reflects a high frequency of intestinal symptoms of sufficient intensity that affect quality of life and disease activity. Early detection of gastrointestinal compromise allows patients to benefit from comprehensive treatment of the disease in its initial stages.

P062 Chromoendoscopy With Magnification Colonoscopy: Analysis of Mucosa of Colon And Ileum In Patients With Gastrointestinal Symptoms Without IBD.

BACKGROUND: Digital chromoendoscopy (Narrow Band Imaging By Olympus) or BLI (Blue Light Imaging By Fujifilm), with the magnification endoscope, allows a detailed evaluation of the mucosal surface and its vascular network, which facilitates the diagnosis and monitoring of early lesions. This technique has demonstrated a better detection, which allows optical diagnosis during a colonoscopy examination. Patients with SpA with nonspecific gastrointestinal symptoms, subclinical intestinal inflammation are defined as endoscopic and histologically. The aim was to detect early structural inflammatory changes by chromoendoscopy and magnification colonoscopy in colonic/ileum digestive mucosa, and establish its association with clinical variables in SpA and gastrointestinal symptoms. Study approved by Institutional Ethics Committee, code HMC 2017-023. METHODS: Clinical evaluation by rheumatologist in SpA patients (ASAS/criteria), fecal calprotectin levels, and HLA-B*27 were evaluated. In patients with ≥2 gastrointestinal symptoms, clinical evaluation by gastroenterologist, digital chromoendoscopy (NBI) or (BLI), magnification colonoscopy, and histological analysis were performed. The association between clinical and colonoscopy variables were established using the Chi-square or Fisher's exact test. RESULTS: In total, 62 SpA patients were included, with mean age of 45.1 ± 11.3 years, axial SpA (77.4%) peripheral SpA (12.9%), biological treatment (69.4%), ASDAS-CRP>2,1 (67.7%), presence of HLA-B*27 (41.9%). Patients with ≥2 gastrointestinal symptoms were found in 67.7%. The most important symptoms were abdominal pain (66.1%), abdominal distension (64.5%), and food intolerance (59.7%). 22.6% of patients showed high level of calprotectin. In those patients with gastrointestinal symptoms, chromoendoscopy and magnification colonoscopy were performed. The mean age of those patients was 45.4 ± 10.5, 57.6% were male, BMI>25 in 69.7%, presence of HLA-B*27 in 39.4%, 33.3% were former smokers, axial SpA in 84.8% and ASDAS-CRP>21 in 78.8%. In total, 27.27% of the patients presented high levels of calprotectin, of which 66.0% had more than two gastrointestinal symptoms (p = 0.015). 77.8% presented alterations in ileal mucosa (p=0.060). The most frequent alteration was the loss of vascular pattern (p = 0.002). By histological analysis, 5 patients had acute inflammation in the ileum, of which 4 had increased levels of fecal calprotectin (p = 0.013). 30.8% of patients positive for HLAB*27:05:02 had ulcers in ileum (p = 0.017) and 61.5% had chronic inflammatory patterns (p=0.020). CONCLUSION: Chromoendoscopy provided an enhanced, detailed contrast of the gastrointestinal mucosa surface, mainly in the loss of vascular pattern in ileum. The active search for symptoms, signs, and biomarkers of gastrointestinal involvement in addition to an objective endoscopic and histological evaluation may offer new perspectives at the evaluation of SpA patients and may provide guidance for specific clinical and therapeutic management.

DNMTs and Impact of CpG Content, Transcription Factors, Consensus Motifs, lncRNAs, and Histone Marks on DNA Methylation.

DNA methyltransferases (DNMTs) play an essential role in DNA methylation and transcriptional regulation in the genome. DNMTs, along with other poorly studied elements, modulate the dynamic DNA methylation patterns of embryonic and adult cells. We summarize the current knowledge on the molecular mechanism of DNMTs' functional targeting to maintain genome-wide DNA methylation patterns. We focus on DNMTs' intrinsic characteristics, transcriptional regulation, and post-transcriptional modifications. Furthermore, we focus special attention on the DNMTs' specificity for target sites, including key cis-regulatory factors such as CpG content, common motifs, transcription factors (TF) binding sites, lncRNAs, and histone marks to regulate DNA methylation. We also review how complexes of DNMTs/TFs or DNMTs/lncRNAs are involved in DNA methylation in specific genome regions. Understanding these processes is essential because the spatiotemporal regulation of DNA methylation modulates gene expression in health and disease.

Generation of an integration-free induced pluripotent stem cell line (UNC001-A) from blood of a healthy individual.

Induced pluripotent stem cells (iPSCs) generated from young, healthy individuals are valuable tools for investigating molecular disease mechanisms during the early development of the brain vasculature. We generated an iPSC line from peripheral blood mononuclear cells (PBMCs) isolated from a healthy 13-yeard old female donor using the Sendai virus. The iPSCs differentiated into endothelial cells, astrocytes, and neurons. This iPSC line can serve as a healthy reference control for comparative studies in drug development and modeling the early onset of Cerebral Cavernous Malformation (CCM).

Caracterización de pacientes con lupus y fiebre: actividad, infección o ambas / Characterisation of patients with lupus and fever: Activity, infection or both

RESUMEN Objetivo: Describir las características clínicas y de laboratorio en pacientes mayores de 15 arios con diagnóstico de lupus eritematoso sistémico (LES) hospitalizados por fiebre, cuyo diagnóstico final fue infección, actividad lúpica o ambas (actividad e infección). Métodos: Se realizó un estudio descriptivo retrospectivo en el que se incluyeron pacientes con diagnóstico de LES admitidos por fiebre en el servicio de urgencias del Hospital Universitario Clínica San Rafael; se estudiaron variables clínicas y paraclínicas, dividiéndose en 3 grupos de interés: pacientes con actividad de la enfermedad, de la infección o de ambas, de acuerdo con el diagnóstico definitivo una vez se daba el alta hospitalaria. Se estudiaron variables clínicas y de laboratorio, realizándose una descripción de la población en los 3 estados. Resultados: Se evaluaron en total 115 pacientes, incluyéndose en el análisis final 108 pacientes. La mediana de edad fue de 36 años y el 86% fueron mujeres. La mediana del puntaje del Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) en toda la población fue de 6 (R 1-15), en los pacientes con infección y actividad el puntaje mostró una mediana de 9,5 (R 6-15). No hubo diferencias significativas entre los síntomas clínicos y los hallazgos de laboratorio en los diferentes grupos. El uso de prednisolona en los últimos 3 meses fue mayor en el grupo de infección (p = 0,001), pero sin diferencias significativas entre las otras terapias de inmunosupresión. Conclusión: El uso de esteroides en los últimos 3 meses, el puntaje de la escala SLEDAI y el tiempo transcurrido desde el diagnóstico de LES podrían ser variables que pueden ayudar a discriminar entre infección y actividad en pacientes con historia de LES y fiebre. Los hallazgos clínicos y paraclínicos no discriminan entre estas condiciones de enfermedad.

ABSTRACT Objective: To describe clinical characteristics and laboratory findings of patients older than 15 years of age diagnosed with Systemic Lupus Erythematosus (SLE) that were admitted to hospital with fever and with a final diagnosis of infection, disease flare-up, or both. Methods: A retrospective chart review of medical charts was performed to identify patients with a diagnosis of SLE with a fever, admitted to the Emergency Department of Hospital Universitario Clinica San Rafael. The data collected included demographics, SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) scores, clinical symptoms, laboratory findings, and steroid use. Results analysis was based on three groups (according to final diagnosis at discharge): patients diagnosed with SLE and infection, SLE and disease flare-up, and SLE with both infection and disease flare-up. Results: This study included 108 patients with a mean age of 36, of whom 86% were female. The mean SLEDAI score of all patients analysed was 6 (R 1-15), and for patients with SLE with both infection and disease flare-up was 9.5 (R 6-15). No significant differences were found with clinical symptoms and laboratory findings for any group analysed. SLE patients diagnosed with infections took significantly more prednisolone in the last three months before admission (P = .001). No differences were found with other immunosuppressants. Conclusion: The use of steroids in the last three months, the SLEDAI score, and time since a SLE diagnosis may be variables used to distinguish between infection and disease flare-up in SLE patients presenting with fever. Clinical symptoms and laboratory results are not useful markers to make this distinction.