Promoting early neovascularization by allotransplanted adipose-derived Muse cells in an ovine model of acute myocardial infarction.

BACKGROUND: Recent preclinical studies have demonstrated that bone marrow (BM)-derived Muse cells have a homing mechanism to reach damaged cardiac tissue while also being able to reduce myocardial infarct size and improve cardiac function; however, the potential of BM-Muse cells to foster new blood-vessel formation has not been fully assessed. Up to date, adipose tissue (AT)-derived Muse cells remain to be studied in acute myocardial infarction (AMI). The aim of the present study was to analyze in vitro and in vivo the neovascularization capacity of AT-Muse cells while exploring their biodistribution and differentiation potential in a translational ovine model of AMI. METHODS AND RESULTS: AT-Muse cells were successfully isolated from ovine adipose tissue. In adult sheep, one or more diagonal branches of the left anterior descending coronary artery were permanently ligated for thirty minutes. Sheep were randomized in two groups and treated with intramyocardial injections: Vehicle (PBS, n = 4) and AT-Muse (2x107 AT-Muse cells labeled with PKH26 Red Fluorescent Dye, n = 4). Molecular characterization showed higher expression of angiogenic genes (VEGF, PGF and ANG) and increased number of tube formation in AT-Muse cells group compared to Adipose-derived mesenchymal stromal cells (ASCs) group. At 7 days post-IAM, the AT-Muse group showed significantly more arterioles and capillaries than the Vehicle group. Co-localization of PKH26+ cells with desmin, sarcomeric actin and troponin T implied the differentiation of Muse cells to a cardiac fate; moreover, PKH26+ cells also co-localized with a lectin marker, suggesting a possible differentiation to a vascular lineage. CONCLUSION: Intramyocardially administered AT-Muse cells displayed a significant neovascularization activity and survival capacity in an ovine model of AMI.

In silico performance analysis of web tools for CRISPRa sgRNA design in human genes.

Angiogenic gene overexpression has been the main strategy in numerous vascular regenerative gene therapy projects. However, most have failed in clinical trials. CRISPRa technology enhances gene overexpression levels based on the identification of sgRNAs with maximum efficiency and safety. CRISPick and CHOP CHOP are the most widely used web tools for the prediction of sgRNAs. The objective of our study was to analyze the performance of both platforms for the sgRNA design to angiogenic genes (VEGFA, KDR, EPO, HIF-1A, HGF, FGF, PGF, FGF1) involving different human reference genomes (GRCH 37 and GRCH 38). The top 20 ranked sgRNAs proposed by the two tools were analyzed in different aspects. No significant differences were found on the DNA curvature associated with the sgRNA binding sites but the sgRNA predicted on-target efficiency was significantly greater when CRISPick was used. Moreover, the mean ranking variation was greater for the same platform in EPO, EGF, HIF-1A, PGF and HGF, whereas it did not reach statistical significance in KDR, FGF-1 and VEGFA. The rearrangement analysis of the ranking positions was also different between platforms. CRISPick proved to be more accurate in establishing the best sgRNAs in relation to a more complete genome, whereas CHOP CHOP showed a narrower classification reordering.

Gene Therapy: Targeting Cardiomyocyte Proliferation to Repopulate the Ischemic Heart.

ABSTRACT: Adult mammalian cardiomyocytes show scarce division ability, which makes the heart ineffective in replacing lost contractile cells after ischemic cardiomyopathy. In the past decades, there have been increasing efforts in the search for novel strategies to regenerate the injured myocardium. Among them, gene therapy is one of the most promising ones, based on recent and emerging studies that support the fact that functional cardiomyocyte regeneration can be accomplished by the stimulation and enhancement of the endogenous ability of these cells to achieve cell division. This capacity can be targeted by stimulating several molecules, such as cell cycle regulators, noncoding RNAs, transcription, and metabolic factors. Therefore, the proposed target, together with the selection of the vector used, administration route, and the experimental animal model used in the development of the therapy would determine the success in the clinical field.

Effect of intramuscular baculovirus encoding mutant hypoxia-inducible factor 1-alpha on neovasculogenesis and ischemic muscle protection in rabbits with peripheral arterial disease.

BACKGROUND AIMS: Peripheral arterial disease (PAD) is a progressive, disabling ailment for which no effective treatment exists. Gene therapy-mediated neovascularization has emerged as a potentially useful strategy. We tested the angiogenic and arteriogenic efficacy and safety of a baculovirus (BV) encoding mutant, oxygen-resistant hypoxia-inducible factor 1-alpha (mHIF-1α), in rabbits with PAD. METHODS: After assessing the transfection efficiency of the BV.mHIF-1α vector and its tubulogenesis potential in vitro, we randomized rabbits with experimental PAD to receive 1 × 109 copies of BV.mHIF-1α or BV.null (n = 6 per group) 7 days after surgery. Two weeks post-treatment, collateralization (digital angiography) and capillary and arteriolar densities (immunohistochemistry) were measured in the posterior limbs. Ischemic damage was evaluated in adductor and gastrocnemius muscle samples. Tracking of viral DNA in injected zones and remote tissues at different time points was performed in additional rabbits using a BV encoding GFP. RESULTS: Angiographically visible collaterals were more numerous in BV.mHIF-1α-treated rabbits (8.12 ± 0.42 vs 6.13 ± 1.15 collaterals/cm2, P < 0.05). The same occurred with arteriolar (27.9 ± 7.0 vs 15.3 ± 4.0 arterioles/mm2) and capillary (341.8 ± 109.9 vs 208.8 ± 87.7 capillaries/mm2, P < 0.05) densities. BV.mHIF-1α-treated rabbits displayed less ischemic muscle damage than BV.null-treated animals. Viral DNA and GFP mRNA were detectable only at 3 and 7 days after injection in hind limbs. Neither the virus nor GFP mRNA was detected in remote tissues. CONCLUSIONS: In rabbits with PAD, BV.mHIF-1α induced neovascularization and reduced ischemic damage, exhibiting a good safety profile at 14 days post-treatment. Complementary studies to evaluate its potential usefulness in the clinic are needed.

Novel insights into cardiac regeneration based on differential fetal and adult ovine heart transcriptomic analysis.

The adult mammalian cardiomyocyte has a very limited capacity to reenter the cell cycle and advance into mitosis. Therefore, diseases characterized by lost contractile tissue usually evolve into myocardial remodeling and heart failure. Analyzing the cardiac transcriptome at different developmental stages in a large mammal closer to the human than laboratory rodents may serve to disclose positive and negative cardiomyocyte cell cycle regulators potentially targetable to induce cardiac regeneration in the clinical setting. Thus we aimed at characterizing the transcriptomic profiles of the early fetal, late fetal, and adult sheep heart by employing RNA-seq technique and bioinformatic analysis to detect protein-encoding genes that in some of the stages were turned off, turned on, or differentially expressed. Genes earlier proposed as positive cell cycle regulators such as cyclin A, cdk2, meis2, meis3, and PCNA showed higher expression in fetal hearts and lower in AH, as expected. In contrast, genes previously proposed as cell cycle inhibitors, such as meis1, p16, and sav1, tended to be higher in fetal than in adult hearts, suggesting that these genes are involved in cell processes other than cell cycle regulation. Additionally, we described Gene Ontology (GO) enrichment of different sets of genes. GO analysis revealed that differentially expressed gene sets were mainly associated with metabolic and cellular processes. The cell cycle-related genes fam64a, cdc20, and cdk1, and the metabolism-related genes pitx and adipoq showed strong differential expression between fetal and adult hearts, thus being potent candidates to be targeted in human cardiac regeneration strategies.NEW & NOTEWORTHY We characterized the transcriptomic profiles of the fetal and adult sheep hearts employing RNAseq technique and bioinformatic analyses to provide sets of transcripts whose variation in expression level may link them to a specific role in cell cycle regulation. It is important to remark that this study was performed in a large mammal closer to humans than laboratory rodents. In consequence, the results can be used for further translational studies in cardiac regeneration.

Functional analysis of six uncharacterised mutations in LDLR gene.

BACKGROUND AND AIMS: Familial hypercholesterolemia (FH) is a primary hyperlipemia. It is an autosomal dominant genetic disorder of lipoproteins metabolism mainly caused by mutations in the low density lipoprotein receptor gene (LDLR). We aimed to investigate the functional impact on the low density lipoprotein receptor (LDLR) activity of six uncharacterised variants located in the coding region of the LDLR gene, namely c.428G > T, c.640T > C, c.1708C > T, c.1736A > T, c.1981C > G and c.2114C > G (NM_000527.4) and to attempt to define their clinical status. METHODS: Functional studies were carried out using site-directed mutagenesis techniques and expression of LDLR protein in vitro. Results were correlated with clinical data and in silico analyses in order to assess the physiopathological role of these variants. RESULTS: This work provides functional information about 6 uncharacterised mutations in LDLR. CONCLUSIONS: The six variants studied here appeared to affect the LDLR function in vitro to different degrees, ranging from receptors with normal to slightly reduced activity to receptors exhibiting less than 10% of the wild-type activity. According to these studies and The American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines, two variants could be classified as "Likely Benign" (p.(Ala705Gly) and p.(Leu570Phe)), three variants as "Pathogenic" (p.(Asp579Val), p.(Cys143Phe) and p.(Trp214Arg)) and one variant as "Likely Pathogenic" (p.(Pro661Ala)).

High-dose intramyocardial HMGB1 induces long-term cardioprotection in sheep with myocardial infarction.

In rodents with acute myocardial infarction (AMI), high mobility group box 1 (HMGB1) injection has produced controversial results. Given the lack of data in large mammals, we searched the dose that would promote angiogenesis and expression of specific regenerative genes in sheep with AMI (protocol 1) and, subsequently, use this dose to study long-term effects on infarct size and left ventricular (LV) function (protocol 2). Protocol 1: Sheep with AMI received 250 µg (high-dose, n = 7), 25 µg (low-dose, n = 7) HMGB1, or PBS (placebo, n = 7) in 10 intramyocardial injections (0.2 ml each) in the peri-infarct area. Seven days later, only the high-HMGB1-dose group exhibited higher microvascular densities, Ki67-positive cardiomyocytes, and overexpression of VEGF, Ckit, Tbx20, Nkx2.5, and Gata4. Protocol 2: Sheep with AMI received HMGB1 250 µg (n = 6) or PBS (n = 6). At 60 days, HMGB1-treated sheep showed smaller infarcts (8.5 ± 2.11 vs. 12.2 ± 1.97% LV area, P < 0.05, ANOVA-Bonferroni) and higher microvascular density (capillaries, 1798 ± 252 vs. 1266 ± 250/mm2; arterioles, 18.3 ± 3.9 vs. 11.7 ± 2.2/mm2; both P < 0.01). Echocardiographic LV ejection fraction, circumferential shortening, and wall thickening increased from day 3 to 60 with HMGB1 (all P < 0.05). Conclusion: in ovine AMI, high-dose HMGB1 induces angio-arteriogenesis, reduces infarct size, and improves LV function at 2 months post-treatment.

Effect of poly (l-lactic acid) scaffolds seeded with aligned diaphragmatic myoblasts overexpressing connexin-43 on infarct size and ventricular function in sheep with acute coronary occlusion.

Diaphragmatic myoblasts (DM) are stem cells of the diaphragm, a muscle displaying high resistance to stress and exhaustion. We hypothesized that DM modified to overexpress connexin-43 (cx43), seeded on aligned poly (l-lactic acid) (PLLA) sheets would decrease infarct size and improve ventricular function in sheep with acute myocardial infarction (AMI). Sheep with AMI received PLLA sheets without DM (PLLA group), sheets with DM (PLLA-DM group), sheets with DM overexpressing cx43 (PLLA-DMcx43) or no treatment (control group, n = 6 per group). Infarct size (cardiac magnetic resonance) decreased ∼25% in PLLA-DMcx43 [from 8.2 ± 0.6 ml (day 2) to 6.5 ± 0.7 ml (day 45), p < .01, ANOVA-Bonferroni] but not in the other groups. Ejection fraction (EF%) (echocardiography) at 3 days post-AMI fell significantly in all groups. At 45 days, PLLA-DM y PLLA-DMcx43 recovered their EF% to pre-AMI values (PLLA-DM: 61.1 ± 0.5% vs. 58.9 ± 3.3%, p = NS; PLLA-DMcx43: 64.6 ± 2.9% vs. 56.9 ± 2.4%, p = NS), but not in control (56.8 ± 2.0% vs. 43.8 ± 1.1%, p < .01) and PLLA (65.7 ± 2.1% vs. 56.6 ± 4.8%, p < .01). Capillary density was higher (p < .05) in PLLA-DMcx43 group than in the remaining groups. In conclusion, PLLA-DMcx43 reduces infarct size in sheep with AMI. PLLA-DMcx43 and PLLA-DM improve ventricular function similarly. Given its safety and feasibility, this novel approach may prove beneficial in the clinic.

Estudio genético de hipercolesterolemia familiar en una población hospitalaria de la Ciudad de Buenos Aires / Genetic Testing of Familial Hypercholesterolemia in a Hospital-Based Population of the City of Buenos Aires

RESUMEN: Introducción: La hipercolesterolemia familiar es una hiperlipidemia primaria. Se trata de un trastorno genético autosómico dominante del metabolismo de las lipoproteínas, caracterizado por concentraciones plasmáticas elevadas de colesterol unido a lipoproteínas de baja densidad y presencia de xantomas tendinosos, y está asociado con el desarrollo prematuro de enfermedad cardiovascular. Objetivos: Investigar la presencia de mutaciones en el principal gen asociado al desarrollo de hipercolesterolemia familiar (LDLR) en un grupo de pacientes identificados como "casos índices", de entre aquellos que concurren al Servicio de Lípidos del Hospital Universitario Fundación Favaloro con diagnóstico clínico de hipercolesterolemia familiar. Determinar la composición ancestral de la población estudiada. Material y métodos: Se estudió una población de 38 pacientes con diagnóstico clínico de hipercolesterolemia familiar. La región codificante y las zonas intrónicas adyacentes del gen LDLR se secuenciaron automáticamente por el método de Sanger. Se investigó el componente ancestral de la población estudiada a partir del análisis de 46 marcadores informativos de ancestralidad (AIM-Indel). Resultados: Se identificaron 50 variantes diferentes, de las cuales el 48% se consideraron patogénicas. Se logró establecer una correlación genotipo-gravedad del fenotipo en el 60,5% de los pacientes estudiados. El componente ancestral de la población estudiada fue predominantemente europeo, seguido de un componente nativo-americano y, en menor proporción, africano. Conclusiones: El análisis genético por secuenciación del gen LDLR en pacientes identificados como "casos índices" con diagnóstico clínico de hipercolesterolemia familiar permite correlacionar el dato genético con la gravedad del fenotipo observado clínicamente y efectuar un diagnóstico en cascada en los miembros de la familia que presentan los criterios de inclusión considerados.

ABSTRACT: Background: Familial hypercholesterolemia is a primary hyperlipidemia. It is an autosomal dominant genetic disorder of lipoprotein metabolism, characterized by elevated plasma low-density lipoprotein cholesterol and presence of tendon xanthomas, and is associated with early cardiovascular disease. Objectives: The aim of this study was to investigate the presence of mutations in the main gene associated with the development of familial hypercholesterolemia (LDLR) in a group of patients identified as "index cases" attending the Lipid Clinic of the Hospital Universitario Fundación Favaloro with clinical diagnosis of familial hypercholesterolemia. The ancestral composition of the study population was determined. Methods: We evaluated 38 patients with clinical diagnosis of familial hypercholesterolemia. Mutation screening of the LDLR gene coding regions and adjacent intronic areas was performed using Sanger sequencing. The ancestral component of the study population was investigated using 46 ancestry inference markers (AIM-Indel). Results: Fifty different variants were identified, 48% of which were considered pathogenic. A genotype-phenotype severity correlation was established in 60.5% of the patients evaluated. The ancestral component of the study population was predominantly European, followed by native-American and African in lower proportion. Conclusions: Genetic testing by LDLR gene sequencing in patients identified as "index cases" with clinical diagnosis of familial hypercholesterolemia allows the correlation between the genetic information and the severity of the clinical phenotype to a cascade testing of the family members presenting the inclusion criteria considered.

Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (L-lactic acid) scaffolds for potential use in cardiac regeneration.

Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion threshold on account that they contract and relax 20 times/min over a lifespan, making them potentially useful in cardiac regeneration strategies. Besides, it has been shown that biomaterials for stem cell delivery improve cell retention and viability in the target organ. In the present study, we aimed at developing a novel approach based on the use of poly (L-lactic acid) (PLLA) scaffolds seeded with DMs overexpressing connexin-43 (cx43), a gap junction protein that promotes inter-cell connectivity. DMs isolated from ovine diaphragm biopsies were characterized by immunohistochemistry and ability to differentiate into myotubes (MTs) and transduced with a lentiviral vector encoding cx43. After confirming cx43 expression (RT-qPCR and Western blot) and its effect on inter-cell connectivity (fluorescence recovery after photobleaching), DMs were grown on fiber-aligned or random PLLA scaffolds. DMs were successfully isolated and characterized. Cx43 mRNA and protein were overexpressed and favored inter-cell connectivity. Alignment of the scaffold fibers not only aligned but also elongated the cells, increasing the contact surface between them. This novel approach is feasible and combines the advantages of bioresorbable scaffolds as delivery method and a cell type that on account of its features may be suitable for cardiac regeneration. Future studies on animal models of myocardial infarction are needed to establish its usefulness on scar reduction and cardiac function.

Combined VEGF gene transfer and erythropoietin in ovine reperfused myocardial infarction.

BACKGROUND: In reperfused acute myocardial infarction (RAMI), cardioprotective treatments may enhance myocardial salvage and hence reduce the area of necrosis. Based on studies showing that plasmid-mediated vascular endothelial growth factor (pVEGF) gene transfer reduces infarct size by combining angio-arteriogenic and cardiomyogenic effects and that erythropoietin (EPO) exerts anti-apoptotic actions in animal models of AMI, we aimed to assess if their association would reduce infarct size to a larger extent than any of them individually in a large mammalian model of RAMI. METHODS: Adult sheep subjected to 90-minute coronary artery occlusion received upon reperfusion intramyocardial pVEGF 3.8 mg plus intravenous EPO 1000 IU/kg (n=8), pVEGF (n=8), EPO (n=8) or placebo (n=8). RESULTS: Fifteen days after treatment, infarct size was smaller in the 3 treatment groups (pVEGF+EPO: 8 ± 1 %; pVEGF: 16 ± 5 %; EPO: 13 ± 4 %) compared to placebo (25 ± 7 %, p<0.001). However, in the EPO+VEGF group infarct size was significantly smaller than in the groups receiving EPO or VEGF individually (p<0.05). DNA fragmentation, a hallmark of late apoptosis, was significantly lower in sheep receiving EPO. The combined treatment, while not affecting global left ventricular performance, improved regional peri-infarct function and prevented over-time expansion of the post-infarct perfusion defect. CONCLUSIONS: Combined pVEGF and EPO treatment might be clinically useful to enhance the benefits of early revascularization in patients with acute myocardial infarction.

Brief report: The potential role of epigenetics on multipotent cell differentiation capacity of mesenchymal stromal cells.

Human umbilical cord perivascular cells (HUCPVCs) are a readily available source of mesenchymal stromal cells (MSCs) for cell therapy. We were interested in understanding how differences from human bone marrow (BM)-derived MSCs might yield insights into MSC biology. We found that HUCPVCs exhibited increased telomerase activity and longer telomeres compared with BM-MSCs. We also observed enhanced expression of the pluripotency factors OCT4, SOX2, and NANOG in HUCPVCs. The methylation of OCT4 and NANOG promoters was similar in both cell types, indicating that differences in the expression of pluripotency factors between the MSCs were not associated with epigenetic changes. MSC methylation at these loci is greater than reported for embryonic stem cells but less than in dermal fibroblasts, suggesting that multipotentiality of MSCs is epigenetically restricted. These results are consistent with the notion that the MSC population (whether BM- or HUCPV-derived) exhibits higher proliferative capacity and contains more progenitor cells than do dermal fibroblasts.

La rosuvastatina atenúa la progresión de la estenosis aórtica generada por hipertensión arterial, independientemente de sus efectos hipolipemiantes / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects

Introducción Existe evidencia epidemiológica que vincula factores de riesgo cardiovascular con la estenosis valvular aórtica. Recientemente se ha demostrado el desarrollo de estenosis valvular aórtica en un modelo de hipertensión arterial en animales. Planteamos la hipótesis de que el tratamiento con rosuvastatina modifica esta transformación. Objetivo Evaluar el efecto de la rosuvastatina sobre el desarrollo de estenosis valvular aórtica. Material y métodos Se instrumentaron conejos NZ machos (n = 43) con el modelo 1-riñón 1-clip de Goldblatt para generar hipertensión arterial. Los animales fueron aleatorizados a: HT (n = 17) que no recibió otro tratamiento, HT+R (n = 14) tratado con rosuvastatina 2,5 mg/kg/día y HT+R+C (n = 12) tratado con rosuvastatina 2,5 mg/kg/día + suplemento de colesterol dietético para mantener los niveles basales de colesterol plasmático. Un grupo control (GC) fue sometido a cirugía simulada (n = 15). Las características de la válvula aórtica se midieron por ecografía en condiciones basales y a los 3 y a los 6 meses de hipertensión arterial. Resultados A los 6 meses de seguimiento, los incrementos de PAS y PAD fueron más elevados en HT (49% y 40%, respectivamente; p < 0,001) en comparación con los grupos tratados con rosuvastatina (PAS = 23% y 25%; PAD = 28% y 26%; p < 0,001 para HT+R y HT+R+C, respectivamente). El colesterol total se redujo el 45,7% (p < 0,01) sólo en HT+R. El espesor valvar se incrementó en HT (0,50 ± 0,01 vs. 0,62 ± 0,02 mm; p < 0,01), sin mostrar diferencias en HT+R y HT+R+C. Finalmente, el área valvular aórtica mostró una reducción en HT (0,277 ± 0,024 vs. 0,208 ± 0,014 cm²; p < 0,05), sin cambios en HT+R y HT+R+C y un aumento no significativo en el GC (0,264 ± 0,022 vs. 0,32 ± 0,016 cm²; p = 0,07). Conclusiones La rosuvastatina atenúa la progresión de la estenosis valvular aórtica generada por hipertensión arterial. Esta protección podría ser mediada por efectos no hipolipemiantes de estas drogas.

Background There is epidemiological evidence associating cardiovascular risk factors with aortic valve stenosis. The development of aortic valve stenosis has been recently demonstrated in a hypertensive animal model. We hypothesize that treatment with rosuvastatin modifies this transformation. Objective To evaluate the effect of rosuvastatin on the development of aortic valve stenosis. Material and Methods Hypertension was induced in 43 male NZ rabbits by a onekidney, one-clip Goldblatt procedure. The animals were randomly assigned to 3 groups: HT (n=17) without treatment; HT+R (n=14) treated with rosuvastatin 2.5 mg/kg/day and HT+R+C (n=12) treated with rosuvastatin 2.5 mg/kg/day + cholesterol-enriched diet to keep baseline cholesterol levels. A control group (CG) underwent sham surgery (n=15). The characteristics of the aortic valve were measured by echocardiography at baseline, 3 and 6 months after inducing hypertension. Results After 6 months of follow-up, SBP and DBP presented greater increase in the group HT (49% and 40%, respectively; p<0.001) compared to groups treated with rosuvastatin (SBP = 23% and 25%; DBP = 28% and 26%; p <0.001 for HT+R and HT+R+C, respectively). Total cholesterol decreased by 45.7% (p <0.01) only in HT+R group. The aortic valve became thickened in the HT group (0.50 ± 0.01 vs. 0.62± 0.02 mm; p <0.01); there were no significant differences in HT+R and HT+R+C. Finally, the aortic valve area was reduced in HT (0.277 ± 0.024 vs. 0.208 ± 0.014 cm² ; p <0.05), had no differences in HT+R and HT+R+C, and presented a non-significant increase in CG (0.264 ± 0.022 vs. 0.32± 0.016 cm²; p=0.07). Conclusions Rosuvastatin slows the progression of aortic valve stenosis caused by hypertension. This protection might be independent of the lipid-lowering effect of the drug.

La rosuvastatina atenúa la progresión de la estenosis aórtica generada por hipertensión arterial, independientemente de sus efectos hipolipemiantes / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects

Introducción Existe evidencia epidemiológica que vincula factores de riesgo cardiovascular con la estenosis valvular aórtica. Recientemente se ha demostrado el desarrollo de estenosis valvular aórtica en un modelo de hipertensión arterial en animales. Planteamos la hipótesis de que el tratamiento con rosuvastatina modifica esta transformación. Objetivo Evaluar el efecto de la rosuvastatina sobre el desarrollo de estenosis valvular aórtica. Material y métodos Se instrumentaron conejos NZ machos (n = 43) con el modelo 1-riñón 1-clip de Goldblatt para generar hipertensión arterial. Los animales fueron aleatorizados a: HT (n = 17) que no recibió otro tratamiento, HT+R (n = 14) tratado con rosuvastatina 2,5 mg/kg/día y HT+R+C (n = 12) tratado con rosuvastatina 2,5 mg/kg/día + suplemento de colesterol dietético para mantener los niveles basales de colesterol plasmático. Un grupo control (GC) fue sometido a cirugía simulada (n = 15). Las características de la válvula aórtica se midieron por ecografía en condiciones basales y a los 3 y a los 6 meses de hipertensión arterial. Resultados A los 6 meses de seguimiento, los incrementos de PAS y PAD fueron más elevados en HT (49% y 40%, respectivamente; p < 0,001) en comparación con los grupos tratados con rosuvastatina (PAS = 23% y 25%; PAD = 28% y 26%; p < 0,001 para HT+R y HT+R+C, respectivamente). El colesterol total se redujo el 45,7% (p < 0,01) sólo en HT+R. El espesor valvar se incrementó en HT (0,50 ± 0,01 vs. 0,62 ± 0,02 mm; p < 0,01), sin mostrar diferencias en HT+R y HT+R+C. Finalmente, el área valvular aórtica mostró una reducción en HT (0,277 ± 0,024 vs. 0,208 ± 0,014 cm²; p < 0,05), sin cambios en HT+R y HT+R+C y un aumento no significativo en el GC (0,264 ± 0,022 vs. 0,32 ± 0,016 cm²; p = 0,07). Conclusiones La rosuvastatina atenúa la progresión de la estenosis valvular aórtica generada por hipertensión arterial. Esta protección podría ser mediada por efectos no hipolipemiantes de estas drogas.(AU)

Background There is epidemiological evidence associating cardiovascular risk factors with aortic valve stenosis. The development of aortic valve stenosis has been recently demonstrated in a hypertensive animal model. We hypothesize that treatment with rosuvastatin modifies this transformation. Objective To evaluate the effect of rosuvastatin on the development of aortic valve stenosis. Material and Methods Hypertension was induced in 43 male NZ rabbits by a onekidney, one-clip Goldblatt procedure. The animals were randomly assigned to 3 groups: HT (n=17) without treatment; HT+R (n=14) treated with rosuvastatin 2.5 mg/kg/day and HT+R+C (n=12) treated with rosuvastatin 2.5 mg/kg/day + cholesterol-enriched diet to keep baseline cholesterol levels. A control group (CG) underwent sham surgery (n=15). The characteristics of the aortic valve were measured by echocardiography at baseline, 3 and 6 months after inducing hypertension. Results After 6 months of follow-up, SBP and DBP presented greater increase in the group HT (49% and 40%, respectively; p<0.001) compared to groups treated with rosuvastatin (SBP = 23% and 25%; DBP = 28% and 26%; p <0.001 for HT+R and HT+R+C, respectively). Total cholesterol decreased by 45.7% (p <0.01) only in HT+R group. The aortic valve became thickened in the HT group (0.50 ± 0.01 vs. 0.62± 0.02 mm; p <0.01); there were no significant differences in HT+R and HT+R+C. Finally, the aortic valve area was reduced in HT (0.277 ± 0.024 vs. 0.208 ± 0.014 cm² ; p <0.05), had no differences in HT+R and HT+R+C, and presented a non-significant increase in CG (0.264 ± 0.022 vs. 0.32± 0.016 cm²; p=0.07). Conclusions Rosuvastatin slows the progression of aortic valve stenosis caused by hypertension. This protection might be independent of the lipid-lowering effect of the drug.(AU)

La rosuvastatina atenúa la progresión de la estenosis aórtica generada por hipertensión arterial, independientemente de sus efectos hipolipemiantes / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects / Rosuvastatin Slows the Progression of Aortic Stenosis Caused by Hypertension Regardless of Its Lipid-Lowering Effects

Introducción Existe evidencia epidemiológica que vincula factores de riesgo cardiovascular con la estenosis valvular aórtica. Recientemente se ha demostrado el desarrollo de estenosis valvular aórtica en un modelo de hipertensión arterial en animales. Planteamos la hipótesis de que el tratamiento con rosuvastatina modifica esta transformación. Objetivo Evaluar el efecto de la rosuvastatina sobre el desarrollo de estenosis valvular aórtica. Material y métodos Se instrumentaron conejos NZ machos (n = 43) con el modelo 1-riñón 1-clip de Goldblatt para generar hipertensión arterial. Los animales fueron aleatorizados a: HT (n = 17) que no recibió otro tratamiento, HT+R (n = 14) tratado con rosuvastatina 2,5 mg/kg/día y HT+R+C (n = 12) tratado con rosuvastatina 2,5 mg/kg/día + suplemento de colesterol dietético para mantener los niveles basales de colesterol plasmático. Un grupo control (GC) fue sometido a cirugía simulada (n = 15). Las características de la válvula aórtica se midieron por ecografía en condiciones basales y a los 3 y a los 6 meses de hipertensión arterial. Resultados A los 6 meses de seguimiento, los incrementos de PAS y PAD fueron más elevados en HT (49% y 40%, respectivamente; p < 0,001) en comparación con los grupos tratados con rosuvastatina (PAS = 23% y 25%; PAD = 28% y 26%; p < 0,001 para HT+R y HT+R+C, respectivamente). El colesterol total se redujo el 45,7% (p < 0,01) sólo en HT+R. El espesor valvar se incrementó en HT (0,50 ñ 0,01 vs. 0,62 ñ 0,02 mm; p < 0,01), sin mostrar diferencias en HT+R y HT+R+C. Finalmente, el área valvular aórtica mostró una reducción en HT (0,277 ñ 0,024 vs. 0,208 ñ 0,014 cm²; p < 0,05), sin cambios en HT+R y HT+R+C y un aumento no significativo en el GC (0,264 ñ 0,022 vs. 0,32 ñ 0,016 cm²; p = 0,07). Conclusiones La rosuvastatina atenúa la progresión de la estenosis valvular aórtica generada por hipertensión arterial. Esta protección podría ser mediada por efectos no hipolipemiantes de estas drogas.(AU)

Background There is epidemiological evidence associating cardiovascular risk factors with aortic valve stenosis. The development of aortic valve stenosis has been recently demonstrated in a hypertensive animal model. We hypothesize that treatment with rosuvastatin modifies this transformation. Objective To evaluate the effect of rosuvastatin on the development of aortic valve stenosis. Material and Methods Hypertension was induced in 43 male NZ rabbits by a onekidney, one-clip Goldblatt procedure. The animals were randomly assigned to 3 groups: HT (n=17) without treatment; HT+R (n=14) treated with rosuvastatin 2.5 mg/kg/day and HT+R+C (n=12) treated with rosuvastatin 2.5 mg/kg/day + cholesterol-enriched diet to keep baseline cholesterol levels. A control group (CG) underwent sham surgery (n=15). The characteristics of the aortic valve were measured by echocardiography at baseline, 3 and 6 months after inducing hypertension. Results After 6 months of follow-up, SBP and DBP presented greater increase in the group HT (49% and 40%, respectively; p<0.001) compared to groups treated with rosuvastatin (SBP = 23% and 25%; DBP = 28% and 26%; p <0.001 for HT+R and HT+R+C, respectively). Total cholesterol decreased by 45.7% (p <0.01) only in HT+R group. The aortic valve became thickened in the HT group (0.50 ñ 0.01 vs. 0.62ñ 0.02 mm; p <0.01); there were no significant differences in HT+R and HT+R+C. Finally, the aortic valve area was reduced in HT (0.277 ñ 0.024 vs. 0.208 ñ 0.014 cm² ; p <0.05), had no differences in HT+R and HT+R+C, and presented a non-significant increase in CG (0.264 ñ 0.022 vs. 0.32ñ 0.016 cm²; p=0.07). Conclusions Rosuvastatin slows the progression of aortic valve stenosis caused by hypertension. This protection might be independent of the lipid-lowering effect of the drug.(AU)

Atherogenic alterations in hypertriglyceridemic patients would not depend on insulin resistance.

BACKGROUND AND AIMS: Several studies have been carried out to characterize the different alterations associated with hypertriglyceridemia (HTG) and to identify this dyslipemia as an independent risk factor for cardiovascular disease (CVD). HTG is frequently, but not always, associated with insulin resistance (IR). The present study was aimed to evaluate if the alterations observed in biomarkers of CVD were similar in HTG states independently of IR. METHODS: HTG was defined as triglycerides ≥1.69 mmol/l and IR as HOMA-IR ≥3.1. HTG-IR patients (n=15) were compared with HTG subjects without IR (WIR) (n=15) and with normotriglyceridemic (NTG)-WIR individuals (n=30). RESULTS: Both HTG groups shared the increment in VLDL-C and non-HDL-C, HDL enrichment in triglycerides and depletion in phospholipids, the decrease in adiponectin concentration, and the increase in CETP activity. HDL-C and VCAM-1 levels were altered only in HTG-IR patients in comparison with the other groups, while oxidized LDL was only higher in HTG-IR than the control group. Multiple regression analysis identified triglycerides as the independent predictor of HDL-C, CETP activity and oxidized LDL levels. CONCLUSION: The increase in triglycerides is the major determinant factor of the atherogenic modifications observed, while IR would be an amplifier factor.

Seguimiento de pacientes sometidos a cirugía cardiovascular: impacto de la educación en la prevención secundaria / Follow-up of patients submitted to cardiovascular surgery: impact of teaching on the secondary prevention

Cardiac rehabilitation programs (CRP), include exercise training, medical advice and education related to cardiovascular pathologies, psychosocial support and behavioral characteristics. This approach shows a sustained positive impact over the cardiovascular risk factors, the physical training and the health-related quality of life of the patients and on adherence to dietary recommendations. During the 2004 the authors started the implementation of the Integral Teaching Program (ITP), focused to convalescent patients after a cardiovascular event and with the purpose to enter in a plan of cardiac rehabilitation and secondary preventions with exercise training programs and with a multidisciplinary approach. The aim of this report was to describe the results of the ITP in a cohort of patients submitted to cardiovascular surgery. In a second term, the impact of the ITP will be measured including the total and partial adherence to the different planes of cardiac rehabilitation programs and the management of vascular risk factors. Also it was studied the improvement in physiological outcomes and health-related quality of life in patients with acute myocardial infarction.

Failure of IL-8 to assess early reperfusion injury following lung transplantation of cardiac death donor pigs.

Although interleukins (IL) 8 and 10 predict lung viability in lung transplantation from heart beating donors (HBD) and IL-1beta is a marker of ex vivo performance from after cardiac death donors (ACDD), IL expression in the recipient remains unknown. This study assessed IL-1beta, IL-8 and IL-10 as indicators of functional performance in single-lung transplantation from ACDD pigs. Animals were divided into: (i) HBD: immediate lung excision; (ii) ACDD: fibrillation, 30 min warm ischemia and 3 h topical cooling. Left lungs of both groups were then flushed with Perfadex and stored at 3-4 degrees C for 3 h. IL in bronchoalveolar lavage fluid (BAL) and hemodynamic and graft function were measured in the donor and during the 2 h reperfusion period in the recipient. Myeloperoxidase, nuclear factor kappa beta, wet/dry weight ratio and a histologic injury score were assessed from biopsies in basal conditions in the donor and at the end of reperfusion. Despite similar pulmonary function and histologic markers of injury in both groups and higher IL-1beta in the donor of ACDD, IL-8 during reperfusion was significantly lower in ACDD (119 +/- 33% of basal) than in HBD (306 +/- 238%, P < 0.05) recipients. The paradoxical behavior of IL-8 makes it an unreliable predictor of ACDD early outcome in this transplantation model.

Categorización del síndrome metabólico de acuerdo con las variables que lo constituyen y su relación con la insulino-resistencia / Consensus definition of the metabolic syndrome and diagnostic criteria for identification and its relationshipo with the insulin-resistance syndrome

In recent years, the cardiovascular prevention has found a new clinical tool: the metabolic syndrome (MS). Arised in the observation of associations between risk factors, it is recognized as the clinical manifestation of insulin resistance. In the course of time, it was demonstrated that the presence of MS possess a prognostic value to predict death, nonfatal acute myocardial infarction, vascular stroke and the development of diabetes... The relationshipo to insulin resistance and diagnostic criteria for identification of the metabolic syndrome are given in the article.

Biomarcadores de aterosclerosis e indicadores de resistencia insulínica en pacientes acromegálicos no diabéticos / Biomarkers of Atherosclerosis and Indicators of Insulin Resistance in Non-Diabetic Acromegalic Patients

Introducción En la acromegalia, las comorbilidades cardiovasculares, respiratorias y metabólicas contribuyen a un aumento significativo de la mortalidad de los pacientes afectados. Asimismo, una proporción elevada de estos pacientes presentan diabetes mellitus. Pese a que el hallazgo de un perfil lipídico y lipoproteico anormal en pacientes acromegálicos suele ser habitual, cuando se intenta identificar y/o establecer el grado de modificaciones de parámetros específicos, los resultados son controversiales. Objetivos Evaluar la presencia de biomarcadores de aterosclerosis en pacientes con acromegalia activa no diabéticos y su asociación con la hormona del crecimiento (GH) y el factor de crecimiento similar a la insulina tipo 1 (IGF-1). Material y métodos Se estudiaron 14 pacientes y 14 controles sanos pareados por sexo y edad. Se midieron las concentraciones de GH e IGF-1 por inmunoensayos. Se evaluaron indicadores de resistencia insulínica (glucosa, insulina y HOMA), perfil lipoproteico, niveles plasmáticos de lipoproteínas de baja densidad oxidadas (LDLox), moléculas de adhesión celular vascular 1 (VCAM-1), endotelina-1 y actividad de fosfolipasa A2 asociada con lipoproteínas (LpPLA2). Resultados En comparación con los controles, los pacientes presentaron aumentos de GH (p < 0,05) e IGF-1 (p < 0,001), de los indicadores de resistencia insulínica (insulina p < 0,001; HOMA p < 0,001), triglicéridos (p < 0,05), apo B (p < 0,001), LDLox (117 ± 20 versus 89 ± 23 U/ L; p < 0,05) y endotelina-1 (0,9 ± 0,2 versus 0,7 ± 0,2 pg/ml; p < 0,05). Más aún, la GH y el IGF-1 se asociaron positivamente con (r; p <) insulina (0,40; 0,05 y 0,73; 0,001), HOMA (0,39; 0,05 y 0,74; 0,001), triglicéridos (0,57; 0,05 y 0,64; 0,001), colesterol de lipoproteínas de muy baja densidad (C-VLDL) (0,54; 0,05 y 0,47; 0,05), apo B (0,40; 0,05 y 0,54; 0,05), LDLox (0,59; 0,05 y 0,66; 0,05) y endotelina-1 (0,55; 0,05 y 0,51; 0,05). Conclusiones Los pacientes con acromegalia activa no diabéticos presentaron un estado de resistencia insulínica, así como modificaciones sutiles en el perfil lipoproteico y concentraciones elevadas de LDLox y endotelina-1. Las alteraciones descriptas podrían contribuir a un estado de mayor propensión al desarrollo de enfermedad cardiovascular aterosclerótica, la cual se sumaría a la miocardiopatía específica de la acromegalia.

Background Cardiovascular, respiratory and metabolic comorbidities associated with acromegaly contribute to a significant increase in the mortality of this disease. Many of these patients are also diabetic. Although it is frequent to find abnormal lipid and lipoprotein profiles in patients with acromegaly, controversial outcomes arise in an attempt to identify and/or establish the degree of the modifications of specific parameters. Objectives To assess the presence of biomarkers of atherosclerosis in non-diabetic patients with active acromegaly and its association with growth hormone (GH) and with insulin-like growth factor type 1 (IGF-1). Material and Methods The study included 14 patients and 14 healthy controls, pared by sex and age. Serum concentration of GH and IGF- 1 were determined by immunoassays. Indicators of insulin resistance (glucose, insulin and HOMA) were measured, as well as lipoprotein profile, plasmatic levels of oxidized LDL (oxLDL), vascular cell adhesion molecule 1 (VCAM-1), endothelin-1 and lipoprotein-associated phospholipase A2 activity (LpPLA2). Results Compared to controls, non-diabetic acromegalic patients had increased levels of GH (p<0.05) and IGF-1 (p<0.001), of indicators of insulin resistance (insulin p<0.001; HOMA p<0.001), triglycerides (p<0.05), apo B (p<0.001), oxLDL (117±20 versus 89±23 U/L; p<0.05) and endothelin-1 (0.9±0.2 versus 0.7±0.2 pg/ml; p<0.05). In addition, GH and IGF-1 were positively associated with (r; p <) insulin (0.40; 0.05 and 0.73; 0.001), HOMA (0.39; 0.05 and 0.74; 0.001), triglycerides (0.57; 0.05 and 0.64; 0.001), very low density lipoprotein-cholesterol (VLDL-C) (0.54; 0.05 and 0.47; 0.05), apo B (0.40; 0.05 and 0.54; 0.05), oxLDL (0.59; 0.05 and 0.66; 0.05) and endothelin-1 (0.55; 0.05 and 0.51; 0.05). Conclusions Non-diabetic patients with active acromegaly presented an insulin-resistant status, as well as subtle modifications of lipid profile and increased levels of oxLDL and endothelin- 1. These alterations could explain why these patients are more likely to develop atherosclerotic cardiovascular disease in addition to acromegalic cardiomyopathy.