The MADS-box genes SOC1 and AGL24 antagonize XAL2 functions in Arabidopsis thaliana root development.

MADS-domain transcription factors play pivotal roles in numerous developmental processes in Arabidopsis thaliana. While their involvement in flowering transition and floral development has been extensively examined, their functions in root development remain relatively unexplored. Here, we explored the function and genetic interaction of three MADS-box genes (XAL2, SOC1 and AGL24) in primary root development. By analyzing loss-of-function and overexpression lines, we found that SOC1 and AGL24, both critical components in flowering transition, redundantly act as repressors of primary root growth as the loss of function of either SOC1 or AGL24 partially recovers the primary root growth, meristem cell number, cell production rate, and the length of fully elongated cells of the short-root mutant xal2-2. Furthermore, we observed that the simultaneous overexpression of AGL24 and SOC1 leads to short-root phenotypes, affecting meristem cell number and fully elongated cell size, whereas SOC1 overexpression is sufficient to affect columella stem cell differentiation. Additionally, qPCR analyses revealed that these genes exhibit distinct modes of transcriptional regulation in roots compared to what has been previously reported for aerial tissues. We identified 100 differentially expressed genes in xal2-2 roots by RNA-seq. Moreover, our findings revealed that the expression of certain genes involved in cell differentiation, as well as stress responses, which are either upregulated or downregulated in the xal2-2 mutant, reverted to WT levels in the absence of SOC1 or AGL24.

APOE Christchurch-mimetic therapeutic antibody reduces APOE-mediated toxicity and tau phosphorylation.

INTRODUCTION: We discovered that the APOE3 Christchurch (APOE3Ch) variant may provide resistance to Alzheimer's disease (AD). This resistance may be due to reduced pathological interactions between ApoE3Ch and heparan sulfate proteoglycans (HSPGs). METHODS: We developed and characterized the binding, structure, and preclinical efficacy of novel antibodies targeting human ApoE-HSPG interactions. RESULTS: We found that one of these antibodies, called 7C11, preferentially bound ApoE4, a major risk factor for sporadic AD, and disrupts heparin-ApoE4 interactions. We also determined the crystal structure of a Fab fragment of 7C11 and used computer modeling to predict how it would bind to ApoE. When we tested 7C11 in mouse models, we found that it reduced recombinant ApoE-induced tau pathology in the retina of MAPT*P301S mice and curbed pTau S396 phosphorylation in brains of systemically treated APOE4 knock-in mice. Targeting ApoE-HSPG interactions using 7C11 antibody may be a promising approach to developing new therapies for AD.

Establishment of Skeletal Myogenic Progenitors from Non-Human Primate Induced Pluripotent Stem Cells.

Pluripotent stem (PS) cells enable the scalable production of tissue-specific derivatives with therapeutic potential for various clinical applications, including muscular dystrophies. Given the similarity to human counterparts, the non-human primate (NHP) is an ideal preclinical model to evaluate several questions, including delivery, biodistribution, and immune response. While the generation of human-induced PS (iPS)-cell-derived myogenic progenitors is well established, there have been no data for NHP counterparts, probably due to the lack of an efficient system to differentiate NHP iPS cells towards the skeletal muscle lineage. Here, we report the generation of three independent Macaca fascicularis iPS cell lines and their myogenic differentiation using PAX7 conditional expression. The whole-transcriptome analysis confirmed the successful sequential induction of mesoderm, paraxial mesoderm, and myogenic lineages. NHP myogenic progenitors efficiently gave rise to myotubes under appropriate in vitro differentiation conditions and engrafted in vivo into the TA muscles of NSG and FKRP-NSG mice. Lastly, we explored the preclinical potential of these NHP myogenic progenitors in a single wild-type NHP recipient, demonstrating engraftment and characterizing the interaction with the host immune response. These studies establish an NHP model system through which iPS-cell-derived myogenic progenitors can be studied.

Nosocomial Infections in Adult Patients Supported by Extracorporeal Membrane Oxygenation in a Cardiac Intensive Care Unit.

The use of venoarterial (VA) extracorporeal membrane oxygenation therapy (ECMO) in patients admitted to cardiac intensive care units (CICU) has increased. Data regarding infections in this population are scarce. In this retrospective study, we analyzed the risk factors, outcome, and predictors of in-hospital mortality due to nosocomial infections in patients with ECMO admitted to a single coronary intensive care unit between July 2013 and March 2019 treated with VA-ECMO for >48 h. From 69 patients treated with VA-ECMO >48 h, (median age 58 years), 29 (42.0%) patients developed 34 episodes of infections with an infection rate of 0.92/1000 ECMO days. The most frequent were ventilator-associated pneumonia (57.6%), tracheobronchitis (9.1%), bloodstream infections (9.1%), skin and soft tissue infections (9.1%), and cytomegalovirus reactivation (9.1%). In-hospital mortality was 47.8%, but no association with nosocomial infections was found (p = 0.75). The number of days on ECMO (OR 1.14, 95% CI 1.01-1.30, p = 0.029) and noninfectious complications were higher in the infected patients (OR: 3.8 95% CI = 1.05-14.1). A higher baseline creatinine value (OR: 8.2 95% CI = 1.12-60.2) and higher blood lactate level at 4 h after ECMO initiation (OR: 2.0 95% CI = 1.23-3.29) were significant and independent risk factors for mortality. Conclusions: Nosocomial infections in medical patients treated with VA-ECMO are very frequent, mostly Gram-negative respiratory infections. Preventive measures could play an important role for these patients.

Off-Hours versus Regular-Hours Implantation of Peripheral Venoarterial Extracorporeal Membrane Oxygenation in Patients with Cardiogenic Shock.

BACKGROUND: The "weekend effect" has been associated with worse clinical outcomes. Our aim was to compare off-hours vs. regular-hours peripheral venoarterial extracorporeal membrane oxygenation (VA-ECMO) in cardiogenic shock patients. METHODS: We analyzed in-hospital and 90-day mortality among 147 consecutive patients treated with percutaneous VA-ECMO for medical reasons between July 1, 2013, and September 30, 2022, during regular-hours (weekdays 8:00 a.m.-10:00 p.m.) and off-hours (weekdays 10:01 p.m.-7:59 a.m., weekends, and holidays). RESULTS: The median patient age was 56 years (interquartile range [IQR] 49-64 years) and 112 (72.6%) were men. The median lactate level was 9.6 mmol/L (IQR 6.2-14.8 mmol/L) and 136 patients (92.5%) had a Society for Cardiovascular Angiography and Interventions (SCAI) stage D or E. Cannulation was performed off-hours in 67 patients (45.6%). In-hospital mortality was similar in off-hours and regular hours (55.2% vs. 56.3%, p = 0.901), as was the 90-day mortality (58.2% vs. 57.5%, p = 0.963), length of hospital stay (31 days [IQR 16-65.8 days] vs. 32 days [IQR 18-63 days], p = 0.979), and VA-ECMO related complications (77.6% vs. 70.0%, p = 0.305). CONCLUSIONS: Off-hours and regular-hours percutaneous VA-ECMO implantation in cardiogenic shock of medical cause have similar results. Our results support well-designed 24/7 VA-ECMO implantation programs for cardiogenic shock patients.

Mobile-phone text messaging to promote ideal cardiovascular health in women.

OBJECTIVE: Ideal cardiovascular health (CVH) was developed to promote CVH as a key component of primordial prevention. Mobile short message service (SMS) is useful for improving health behaviours. We aim to test the effectiveness of SMS intervention in women to improve CVH. METHODS: In a single-blinded, randomised, controlled study, 620 women, aged 35-70 years, without cardiovascular disease, were enrolled in SMS intervention versus no SMS. CVH metrics by self-report, and biochemical laboratory, anthropometric and blood pressure measurements were collected during home visits at baseline and 9 months. Women were categorised as having poor (0-2), intermediate (3-4) or ideal (5-7) CVH according to the number of ideal CVH metrics. Participants were randomised 1:1 to SMS intervention versus control. SMS was sent every 5-6 days for 9 months. The primary outcome was the difference in the proportion of women with ideal CVH between SMS and control groups at 9 months. Rates of intermediate CVH, poor CVH and each of the seven ideal CV health metrics at 9 months were key secondary endpoints. RESULTS: At 9 months, there was no significant difference between groups for the primary outcome (16.3% at baseline and 13.3% at 9 months, and 10.1% and 11.1%, in SMS and control groups, respectively, adjusted RR 1.0; 95% CI 0.6 to 1.6). Similarly, there were no significant differences between groups for the key secondary endpoints. SMS had an acceptance rate of 94.9%. CONCLUSIONS: Behavioural SMS intervention did not improve rates of ideal CVH in women, despite being feasible and well received. TRIAL REGISTRATION NUMBER: 6377.

Lactate levels as a prognostic predict in cardiogenic shock under venoarterial extracorporeal membrane oxygenation support.

INTRODUCTION AND OBJECTIVES: Lactate and its evolution are associated with the prognosis of patients in shock, although there is little evidence in those assisted with an extracorporeal venoarterial oxygenation membrane (VA-ECMO). Our objective was to evaluate its prognostic value in cardiogenic shock assisted with VA-ECMO. METHODS: Study of patients with cardiogenic shock treated with VA-ECMO for medical indication between July 2013 and April 2021. Lactate clearance was calculated: [(initial lactate - 6 h lactate) / initial lactate × exact time between both determinations]. RESULTS: From 121 patients, 44 had acute myocardial infarction (36.4%), 42 implant during cardiopulmonary resuscitation (34.7%), 14 pulmonary embolism (11.6%), 14 arrhythmic storm (11.6%), and 6 fulminant myocarditis (5.0%). After 30 days, 60 patients (49.6%) died, mortality was higher for implant during cardiopulmonary resuscitation than for implant in spontaneous circulation (30 of 42 [71.4%] vs 30 of 79 [38.0%], P=.030). Preimplantation GPT and lactate (both baseline, at 6hours, and clearance) were independently associated with 30-day mortality. The regression models that included lactate clearance had a better predictive capacity for survival than the ENCOURAGE and ECMO-ACCEPTS scores, with the area under the ROC curve being greater in the model with lactate at 6 h. CONCLUSIONS: Lactate (at baseline, 6h, and clearance) is an independent predictor of prognosis in patients in cardiogenic shock supported by VA-ECMO, allowing better risk stratification and predictive capacity.

Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies.

Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell-derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell-derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.

A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy.

Mutations in the fukutin-related protein (FKRP) gene result in a broad spectrum of muscular dystrophy (MD) phenotypes, including the severe Walker-Warburg syndrome (WWS). Here, we develop a gene-editing approach that replaces the entire mutant open reading frame with the wild-type sequence to universally correct all FKRP mutations. We apply this approach to correct FKRP mutations in induced pluripotent stem (iPS) cells derived from patients displaying broad clinical severity. Our findings show rescue of functional α-dystroglycan (α-DG) glycosylation in gene-edited WWS iPS cell-derived myotubes. Transplantation of gene-corrected myogenic progenitors in the FKRPP448L-NSG mouse model gives rise to myofiber and satellite cell engraftment and, importantly, restoration of α-DG functional glycosylation in vivo. These findings suggest the potential feasibility of using CRISPR-Cas9 technology in combination with patient-specific iPS cells for the future development of autologous cell transplantation for FKRP-associated MDs.

NAD+ enhances ribitol and ribose rescue of α-dystroglycan functional glycosylation in human FKRP-mutant myotubes.

Mutations in the fukutin-related protein (FKRP) cause Walker-Warburg syndrome (WWS), a severe form of congenital muscular dystrophy. Here, we established a WWS human induced pluripotent stem cell-derived myogenic model that recapitulates hallmarks of WWS pathology. We used this model to investigate the therapeutic effect of metabolites of the pentose phosphate pathway in human WWS. We show that functional recovery of WWS myotubes is promoted not only by ribitol but also by its precursor ribose. Moreover, we found that the combination of each of these metabolites with NAD+ results in a synergistic effect, as demonstrated by rescue of α-dystroglycan glycosylation and laminin binding capacity. Mechanistically, we found that FKRP residual enzymatic capacity, characteristic of many recessive FKRP mutations, is required for rescue as supported by functional and structural mutational analyses. These findings provide the rationale for testing ribose/ribitol in combination with NAD+ to treat WWS and other diseases associated with FKRP mutations.

Healthy muscles are complex machines that require a myriad of finely tuned molecules to work properly. For instance, a protein called alpha-DG sits at the surface of healthy muscle cells, where it strengthens the tissue by latching onto other proteins in the environment. To perform its role correctly, it first needs to be coated with sugar molecules, a complex process which requires over 20 proteins, including the enzyme FKRP. Faulty forms of FKRP reduce the number of sugars added to alpha-DG, causing the muscle tissue to weaken and waste away, potentially leading to severe forms of diseases known as muscular dystrophies. Drugs that can restore alpha-DG sugar molecules could help to treat these conditions. Previous studies on mice and fish have highlighted two potential candidates, known as ribitol and NAD+, which can help to compensate for reduced FKRP activity and allow sugars to be added to alpha-DG again. Yet no model is available to test these molecules on actual human muscle cells. Here, Ortiz-Cordero et al. developed such a model in the laboratory by growing muscle cells from naïve, undifferentiated cells generated from skin given by a muscular dystrophy patient with a faulty form of FKRP. The resulting muscle fibers are in essence identical to the ones present in the individual. As such, they can help to understand the effect various drugs have on muscular dystrophies. The cells were then put in contact with either NAD+, ribitol, or a precursor of ribitol known as ribose. Ortiz-Cordero et al. found that ribitol and ribose restored the ability of FKRP to add sugars to alpha-DG, reducing muscle damage. Combining NAD+ with ribitol or ribose had an even a bigger impact, further increasing the number of sugars on alpha-DG. The human muscle cell model developed by Ortiz-Cordero et al. could help to identify new compounds that can treat muscular conditions. In particular, the findings point towards NAD+, ribose and ribitol as candidates for treating FKRP-related muscular dystrophies. Further safety studies are now needed to evaluate whether these compounds could be used in patients.

Canulación urgente a pie de cama y sin anticoagulación de membrana de oxigenación extracorpórea venovenosa en un paciente con hemoptisis masiva y shock refractario / Emergent Bedside and Anticoagulation-free Veno-venal Extracorporeal Oxygenation Membrane Cannulation in a Patient with Massive Hemoptysis and Unresponsive Shock

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Prevalence and determinants of ideal cardiovascular health in a latin women cohort: a cross-sectional study.

Background: Ideal cardiovascular health (CVH) sought to reduce cardiovascular (CV) morbidity and mortality. In Chile, CV mortality in women is high. The study's main aim was to determine the prevalence of ideal CVH, and the factors and behaviors associated with ideal CVH in women from Santiago de Chile. Methods: Cross-sectional study in women between 35 - 70 years old who were selected through a probabilistic, multistage, and geographically stratified sampling. The study included a survey on demographic and CV risk factors and anthropometric, blood pressure, and biochemical measurements. Three categories were used to characterize low (0-2), intermediate (3-4), and high (5-7) levels of AHA's Ideal CVH index. We assessed the prevalence of ideal CVH by age, education level, and socioeconomic status and determined the independent associations of different variables with ideal CVH. Findings: 620 women, mean age 51± 4 years old, were recruited. Ideal CVH prevalence was 14.3%; none of the women presented an ideal healthy diet, and only 22.6% reached an ideal BMI. The best predictors of ideal CVH were a high education level (OR= 2.85; 1.43 to 5.92; p < 0.01), having less than two alcoholic drinks per day (OR= 4.09; 1.60 to 13.77; p< 0.01), and having a pregnancy history without preeclampsia and/or gestational diabetes (OR=1.94; 1.07 to 3.71; p=0.04). Interpretation: This study demonstrates a low ideal CVH prevalence in Chilean women. Education level was a significant factor associated with ideal CVH. But also, women-specific risk factors, such as a history of preeclampsia/gestational diabetes, and alcohol consumption, were important factors related to CVH. Funding: This study was supported by grants from Fundación SOCHICAR de la Sociedad Chilena de Cardiología y Cirugía Cardiovascular, the American Heart Association and an unrestricted grant by TEVA Pharmaceuticals.

Fukutin-Related Protein: From Pathology to Treatments.

Fukutin-related protein (FKRP) is a glycosyltransferase involved in the functional glycosylation of α-dystroglycan (DG), a key component in the link between the cytoskeleton and the extracellular matrix (ECM). Mutations in FKRP lead to dystroglycanopathies with broad severity, including limb-girdle and congenital muscular dystrophy. Studies over the past 5 years have elucidated the function of FKRP, which has expanded the number of therapeutic opportunities for patients carrying FKRP mutations. These include small molecules, gene delivery, and cell therapy. Here we summarize recent findings on the function of FKRP and describe available models for studying diseases and testing therapeutics. Lastly, we highlight preclinical studies that hold potential for the treatment of FKRP-associated dystroglycanopathies.

Low Cardiovascular Disease Awareness in Chilean Women: Insights from the ESCI Project.

Background: Although cardiovascular disease (CVD) is the leading cause of mortality in Latin American women, limited data exist on CVD perceptions in this population. This study aimed to assess CVD awareness and knowledge of women from Santiago, Chile. Methods: This was a cross-sectional study conducted in women 35 to 70 years old. A multistage probability sampling (stratified by age and socioeconomic level) was used for participant selection. Participants completed a home survey about knowledge of CVD, risk factors, and perceived risk (based on standardized questions from the American Heart Association awareness survey). Results: 723 women participated in the study (mean age: 51 ± 9 years; 17.6% with high education level). Only 9.3% of the respondents mentioned CVD as women's primary health problem, whereas 22.7% and 16.1%, respectively, listed breast cancer and other cancers. When asked to identify the leading cause of women's death, only 14.4% identified CVD compared to 69.1% who recorded cancer. Older women (≥ 55 years) more likely identified CVD as the main cause of death: (OR 2.9: 95% CI = 1.8-4.5) versus younger women (<55 years). CVD family history was also associated with higher awareness of CVD as the leading cause of death (OR 1.7: 95% IC; p = 1.1-2.6). Instead, women with middle education level were less likely to mention CVD as the main women's killer. Conclusions: Chilean women from Santiago have a low awareness of CVD as the leading cause of death and do not recognize CVD as their prominent health problem. Efforts should focus on increasing awareness and knowledge about CVD especially in young women.

Efficient engraftment of pluripotent stem cell-derived myogenic progenitors in a novel immunodeficient mouse model of limb girdle muscular dystrophy 2I.

BACKGROUND: Defects in α-dystroglycan (DG) glycosylation characterize a group of muscular dystrophies known as dystroglycanopathies. One of the key effectors in the α-DG glycosylation pathway is the glycosyltransferase fukutin-related protein (FKRP). Mutations in FKRP lead to a large spectrum of muscular dystrophies, including limb girdle muscular dystrophy 2I (LGMD2I). It remains unknown whether stem cell transplantation can promote muscle regeneration and ameliorate the muscle wasting phenotype associated with FKRP mutations. RESULTS: Here we transplanted murine and human pluripotent stem cell-derived myogenic progenitors into a novel immunodeficient FKRP-mutant mouse model by intra-muscular injection. Upon both mouse and human cell transplantation, we observe the presence of donor-derived myofibers even in absence of pre-injury, and the rescue of α-DG functional glycosylation, as shown by IIH6 immunoreactivity. The presence of donor-derived cells expressing Pax7 under the basal lamina is indicative of satellite cell engraftment, and therefore, long-term repopulation potential. Functional assays performed in the mouse-to-mouse cohort revealed enhanced specific force in transplanted muscles compared to PBS-injected controls. CONCLUSIONS: Altogether, our data demonstrate for the first time the suitability of a cell-based therapeutic approach to improve the muscle phenotype of dystrophic FKRP-mutant mice.

Reanimación cardiopulmonar con ECMO percutáneo en parada cardiaca refractaria hospitalaria: experiencia de un centro / Cardiopulmonary resuscitation with percutaneous ECMO in refractory in-hospital cardiac arrest: a single-center experience

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Uso del oxigenador extracorpóreo de membrana venoarterial en pacientes con tromboembolia pulmonar de alto riesgo / Use of extracorporeal membrane oxygenator in massive pulmonary embolism

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Desarrollo y validación externa de un modelo pronóstico precoz para supervivientes de una parada cardiaca extrahospitalaria / Development and external validation of an early prognostic model for survivors of out-of-hospital cardiac arrest

Introducción y objetivos: A pesar del uso de la hipotermia terapéutica, los pacientes recuperados tras sufrir una parada cardiaca extrahospitalaria tienen un elevado riesgo de muerte o deterioro neurológico grave. Se analizaron la utilidad de diversas variables disponibles al ingreso hospitalario para predecir su evolución a los 6 meses. Métodos: Se desarrolló un estudio multicéntrico en 3 unidades de cuidados intensivos cardiacos. El análisis se realizó sobre 153 pacientes ingresados en dos centros tras sufrir una parada cardiaca extrahospitalaria recuperada y que se trataron con control de temperatura, entre enero de 2007 y julio de 2015. Se consideraron secuelas neurológicas significativas si la Cerebral Performance Categories Scale > 2 a los 6 meses. Los resultados se validaron externamente con los datos procedentes de otros 91 pacientes ingresados en un tercer hospital, durante el mismo periodo de tiempo. Resultados: Del total de 244 pacientes (mediana de edad, 60 años; 77,1% varones; 50,0% en el contexto de isquemia miocárdica aguda), 107 (43,8%) sobrevivieron a los 6 meses con una evolución neurológica favorable. Se calculó un modelo predictivo que incluyó 5 variables (primer ritmo, edad, lactato al ingreso, tiempo hasta recuperación de circulación espontánea y diabetes), con un área bajo la curva de 0,90 (IC95%, 0,85-0,95). Cuando se realizó la validación externa del modelo, la sensibilidad fue de 73,5%, con una especificidad de 78,6% y un área bajo la curva de 0,82 (IC95%, 0,73-0,91). Conclusiones: Un modelo predictivo que incluye cinco variables disponibles en el momento de ingreso de pacientes recuperados tras sufrir una parada cardiaca extrahospitalaria puede ayudar a predecir la probabilidad de supervivencia libre de secuelas neurológicas graves en el seguimiento

Introduction and objectives: Despite therapeutic hypothermia, unconscious survivors of out-of-hospital cardiac arrest have a high risk of death or poor neurologic function. Our objective was to assess the usefulness of the variables obtained in the early moments after resuscitation in the prediction of 6-month prognosis. Methods: A multicenter study was performed in 3 intensive cardiac care units. The analysis was done in 153 consecutive survivors of out-of-hospital cardiac arrest who underwent targeted temperature management between January 2007 and July 2015. Significant neurological sequelae at 6 months were considered to be present in patients with Cerebral Performance Categories Scale > 2. An external validation was performed with data from 91 patients admitted to a third hospital in the same time interval. Results: Among the 244 analyzed patients (median age, 60 years; 77.1% male; 50.0% in the context of acute myocardial ischemia), 107 patients (43.8%) survived with good neurological status at 6 months. The prediction model included 5 variables (Shockable rhythm, Age, Lactate levels, Time Elapsed to return of spontaneous circulation, and Diabetes - SALTED) and provided an area under the curve of 0.90 (95%CI, 0.85-0.95). When external validation was performed, the predictive model showed a sensitivity of 73.5%, specificity of 78.6%, and area under the curve of 0.82 (95%CI, 0.73-0.91). Conclusions: A predictive model that includes 5 clinical and easily accessible variables at admission can help to predict the probability of survival without major neurological damage following out-of-hospital cardiac arrest