Dynamic Single Cell Transcriptomics Defines Kidney FGF23/KL Bioactivity and Novel Segment-Specific Inflammatory Targets.

FGF23 via its coreceptor αKlotho (KL) provides critical control of phosphate metabolism, which is altered in rare and very common syndromes, however the spatial-temporal mechanisms dictating renal FGF23 functions remain poorly understood. Thus, developing approaches to modify specific FGF23-dictated pathways has proven problematic. Herein, wild type mice were injected with rFGF23 for 1, 4 and 12h and renal FGF23 bioactivity was determined at single cell resolution. Computational analysis identified distinct epithelial, endothelial, stromal, and immune cell clusters, with differential expressional analysis uniquely tracking FGF23 bioactivity at each time point. FGF23 actions were sex independent but critically relied upon constitutive KL expression mapped within proximal tubule (S1-S3) and distal tubule (DCT/CNT) cell sub-populations. Temporal KL-dependent FGF23 responses drove unique and transient cellular identities, including genes in key MAPK- and vitamin D-metabolic pathways via early- (AP-1-related) and late-phase (EIF2 signaling) transcriptional regulons. Combining ATACseq/RNAseq data from a cell line stably expressing KL with the in vivo scRNAseq pinpointed genomic accessibility changes in MAPK-dependent genes, including the identification of FGF23-dependent EGR1 distal enhancers. Finally, we isolated unexpected crosstalk between FGF23-mediated MAPK signaling and pro-inflammatory TNF receptor activation via NF-κB, which blocked FGF23 bioactivity in vitro and in vivo . Collectively, our findings have uncovered novel pathways at the single cell level that likely influence FGF23-dependent disease mechanisms. Translational statement: Inflammation and elevated FGF23 in chronic kidney disease (CKD) are both associated with poor patient outcomes and mortality. However, the links between these manifestations and the effects of inflammation on FGF23-mediated mineral metabolism within specific nephron segments remain unclear. Herein, we isolated an inflammatory pathway driven by TNF/NF-κB associated with regulating FGF23 bioactivity. The findings from this study could be important in designing future therapeutic approaches for chronic mineral diseases, including potential combination therapies or early intervention strategies. We also suggest that further studies could explore these pathways at the single cell level in CKD models, as well as test translation of our findings to interactions of chronic inflammation and elevated FGF23 in human CKD kidney datasets.

Inflammation and Oxidative Stress Induced by Obesity, Gestational Diabetes, and Preeclampsia in Pregnancy: Role of High-Density Lipoproteins as Vectors for Bioactive Compounds.

Inflammation and oxidative stress are essential components in a myriad of pathogenic entities that lead to metabolic and chronic diseases. Moreover, inflammation in its different phases is necessary for the initiation and maintenance of a healthy pregnancy. Therefore, an equilibrium between a necessary/pathologic level of inflammation and oxidative stress during pregnancy is needed to avoid disease development. High-density lipoproteins (HDL) are important for a healthy pregnancy and a good neonatal outcome. Their role in fetal development during challenging situations is vital for maintaining the equilibrium. However, in certain conditions, such as obesity, diabetes, and other cardiovascular diseases, it has been observed that HDL loses its protective properties, becoming dysfunctional. Bioactive compounds have been widely studied as mediators of inflammation and oxidative stress in different diseases, but their mechanisms of action are still unknown. Nonetheless, these agents, which are obtained from functional foods, increase the concentration of HDL, TRC, and antioxidant activity. Therefore, this review first summarizes several mechanisms of HDL participation in the equilibrium between inflammation and oxidative stress. Second, it gives an insight into how HDL may act as a vector for bioactive compounds. Third, it describes the relationships between the inflammation process in pregnancy and HDL activity. Consequently, different databases were used, including MEDLINE, PubMed, and Scopus, where scientific articles published in the English language up to 2023 were identified.

Osteocyte Egln1/Phd2 links oxygen sensing and biomineralization via FGF23.

Osteocytes act within a hypoxic environment to control key steps in bone formation. FGF23, a critical phosphate-regulating hormone, is stimulated by low oxygen/iron in acute and chronic diseases, however the molecular mechanisms directing this process remain unclear. Our goal was to identify the osteocyte factors responsible for FGF23 production driven by changes in oxygen/iron utilization. Hypoxia-inducible factor-prolyl hydroxylase inhibitors (HIF-PHI) which stabilize HIF transcription factors, increased Fgf23 in normal mice, as well as in osteocyte-like cells; in mice with conditional osteocyte Fgf23 deletion, circulating iFGF23 was suppressed. An inducible MSC cell line ('MPC2') underwent FG-4592 treatment and ATACseq/RNAseq, and demonstrated that differentiated osteocytes significantly increased HIF genomic accessibility versus progenitor cells. Integrative genomics also revealed increased prolyl hydroxylase Egln1 (Phd2) chromatin accessibility and expression, which was positively associated with osteocyte differentiation. In mice with chronic kidney disease (CKD), Phd1-3 enzymes were suppressed, consistent with FGF23 upregulation in this model. Conditional loss of Phd2 from osteocytes in vivo resulted in upregulated Fgf23, in line with our findings that the MPC2 cell line lacking Phd2 (CRISPR Phd2-KO cells) constitutively activated Fgf23 that was abolished by HIF1α blockade. In vitro, Phd2-KO cells lost iron-mediated suppression of Fgf23 and this activity was not compensated for by Phd1 or -3. In sum, osteocytes become adapted to oxygen/iron sensing during differentiation and are directly sensitive to bioavailable iron. Further, Phd2 is a critical mediator of osteocyte FGF23 production, thus our collective studies may provide new therapeutic targets for skeletal diseases involving disturbed oxygen/iron sensing.

Gastroesophageal reflux disease in patients with long standing type 1 diabetes mellitus: utility of two self-report questionnaires in a multifactorial disease.

BACKGROUND: Gastroesophageal pathologies are common and multifactorial in patients with type 1 diabetes (T1DM). The evaluation with endoscopy and 24 h pH esophageal monitoring is expensive and not always available in all medical centers, especially in developing countries so more cost-effective algorithms for diagnosis are required. Clinical questionnaires are easy to apply but its utility for gastroesophageal reflux disease screening in patients with long standing T1DM must be analyzed. OBJECTIVE: To evaluate the utility of the FSSG and Carlsson-Dent (CDQ) questionnaires to detect the frequency of gastroesophageal reflux disease in patients with T1DM. METHODS: Analytic cross-sectional study, included 54 randomly selected patients from the T1DM clinic in our hospital. Before their routine evaluation, were asked to answer FSSG and CDQ questionnaires, classifying them as positive with a score >8 or >4, respectively. we associated and compared the clinical and biochemical characteristics between patients with or without gastroesophageal reflux detected through questionnaires. RESULTS: Median age was 29 years (22-35), 67% were female (median of 16 years from diagnosis). In 39% of the patients FSSG was positive, CDQ was positive in 28%. A total of 71% of patients were taking medications to treat non-specific gastric symptoms. The concordance between questionnaires was 65% (p: <0.001). Those patients with tobacco consumption as well as those with poor glycemic control were more likely to score positive in either questionnaire. CONCLUSIONS: Patients T1DM had a high prevalence of gastroesophageal reflux disease. In those patients FSSG questionnaire detected a higher number of patients in comparison with CDQ.

INTRODUCCIÓN: Las patologías gastroesofágicas son comunes y multifactoriales en pacientes con diabetes tipo 1 (DM1). La evaluación por medio de panendoscopia y pHmetría es costosa y difícil de realizar en todos los centros de atención, por lo que se requieren algoritmos rentables para su diagnóstico. Existen cuestionarios sencillos y autoaplicables que pueden ser útiles para el diagnóstico de enfermedad por reflujo gastroesofágico en los pacientes con DM1. OBJETIVO: Evaluar la utilidad de los cuestionarios FSSG y Carlsson-Dent (CDQ) para detectar la enfermedad por reflujo gastroesofágico (ERGE) en pacientes con DM1. MÉTODOS: Estudio transversal, se incluyeron 54 pacientes, elegidos al azar de la clínica de DMT1. Previo a la consulta, se les solicitó contestaran los cuestionarios FSSG y el CDQ, considerándose positivos para diagnóstico de ERGE los puntajes >8 y >4, respectivamente. Se analizaron y compararon las características bioquímicas y clínicas entre los pacientes con y sin síntomas de ERGE detectada por medio de los cuestionarios. RESULTADOS: Los pacientes estudiados tenían edad de 29 años (22-35), 67% fueron mujeres, (mediana de diagnóstico de 16 años). El 39% de los pacientes tenían ERGE detectado mediante FSSG y 28% utilizando el cuestionario CDQ. El 71% de los pacientes reportó uso de medicamentos para reflujo. La concordancia entre ambos cuestionarios fue del 65% (p: <0.001). Pacientes que consumen tabaco y con descontrol glucémico, tenían más probabilidades de positividad en cualquier cuestionario. CONCLUSIONES: Existe una alta prevalencia de ERGE en los pacientes con DM1. En esta población el cuestionario FSSG detectó a un mayor número de pacientes en comparación con el CDQ.

Gastroesophageal reflux disease in patients with long standing type 1 diabetes mellitus: utility of two self-report questionnaires in a multifactorial disease / Enfermedad por Reflujo Gastroesofágico en pacientes con Diabetes Mellitus tipo 1: utilidad de dos cuestionarios auto-aplicables

Abstract Background: Gastroesophageal pathologies are common and multifactorial in patients with type 1 diabetes (T1DM). The evaluation with endoscopy and 24 h pH esophageal monitoring is expensive and not always available in all medical centers, especially in developing countries so more cost-effective algorithms for diagnosis are required. Clinical questionnaires are easy to apply but its utility for gastroesophageal reflux disease screening in patients with long standing T1DM must be analyzed. Objective: To evaluate the utility of the FSSG and Carlsson-Dent (CDQ) questionnaires to detect the frequency of gastroesophageal reflux disease in patients with T1DM. Methods: Analytic cross-sectional study, included 54 randomly selected patients from the T1DM clinic in our hospital. Before their routine evaluation, were asked to answer FSSG and CDQ questionnaires, classifying them as positive with a score >8 or >4, respectively. we associated and compared the clinical and biochemical characteristics between patients with or without gastroesophageal reflux detected through questionnaires. Results: Median age was 29 years (22-35), 67% were female (median of 16 years from diagnosis). In 39% of the patients FSSG was positive, CDQ was positive in 28%. A total of 71% of patients were taking medications to treat non-specific gastric symptoms. The concordance between questionnaires was 65% (p: <0.001). Those patients with tobacco consumption as well as those with poor glycemic control were more likely to score positive in either questionnaire. Conclusions: Patients T1DM had a high prevalence of gastroesophageal reflux disease. In those patients FSSG questionnaire detected a higher number of patients in comparison with CDQ.

Resumen Introducción: Las patologías gastroesofágicas son comunes y multifactoriales en pacientes con diabetes tipo 1 (DM1). La evaluación por medio de panendoscopia y pHmetría es costosa y difícil de realizar en todos los centros de atención, por lo que se requieren algoritmos rentables para su diagnóstico. Existen cuestionarios sencillos y autoaplicables que pueden ser útiles para el diagnóstico de enfermedad por reflujo gastroesofágico en los pacientes con DM1. Objetivo: Evaluar la utilidad de los cuestionarios FSSG y Carlsson-Dent (CDQ) para detectar la enfermedad por reflujo gastroesofágico (ERGE) en pacientes con DM1. Métodos: Estudio transversal, se incluyeron 54 pacientes, elegidos al azar de la clínica de DMT1. Previo a la consulta, se les solicitó contestaran los cuestionarios FSSG y el CDQ, considerándose positivos para diagnóstico de ERGE los puntajes >8 y >4, respectivamente. Se analizaron y compararon las características bioquímicas y clínicas entre los pacientes con y sin síntomas de ERGE detectada por medio de los cuestionarios. Resultados: Los pacientes estudiados tenían edad de 29 años (22-35), 67% fueron mujeres, (mediana de diagnóstico de 16 años). El 39% de los pacientes tenían ERGE detectado mediante FSSG y 28% utilizando el cuestionario CDQ. El 71% de los pacientes reportó uso de medicamentos para reflujo. La concordancia entre ambos cuestionarios fue del 65% (p: <0.001). Pacientes que consumen tabaco y con descontrol glucémico, tenían más probabilidades de positividad en cualquier cuestionario. Conclusiones: Existe una alta prevalencia de ERGE en los pacientes con DM1. En esta población el cuestionario FSSG detectó a un mayor número de pacientes en comparación con el CDQ.

Psychosis Crisis Associated with Thyrotoxicosis due to Graves' Disease.

We present the case of a patient with previous psychiatric illness, acutely exacerbated by thyroid storm due to Graves' disease, in whom treatment with antipsychotics induced catatonia. These associations are extremely rare and may be confused with Hashimoto's encephalopathy, especially in the presence of anti-thyroid antibodies in cerebrospinal fluid. The treatment consists in the control of the triggering disease (in this case the resolution of the thyrotoxicosis) and the use of benzodiazepines. However, in some cases, the resolution of psychiatric symptoms is partial and may require the use of electroconvulsive therapy.