Sedoanalgesia para procedimientos en Unidad de Cuidados Intensivos Pediátricos (UCIP). Farmacología, efectos secundarios y control de calidad / Sedoanalgesia for procedures in Pediatric Intensive Care Unit (PICU). Pharmacology, side effects and quality control

OBJETIVOS: Analizar los fármacos más utilizados para sedoanalgesia en procedimientos realizados en una Unidad de Cuidados Intensivos Pediátricos (UCIP), medir su efectividad (nivel de sedación), efectos secundarios y control de calidad. MATERIAL Y MÉTODOS: Estudio prospectivo, observacional y descriptivo. Se recogieron datos epidemiológicos y clínicos, fármaco/s utilizado/s, nivel de sedación alcanzado, incidencias o efectos adversos y escalas de satisfacción, de pacientes de 0 a 18 años sometidos a procedimientos que precisaron sedoanalgesia. RESULTADOS: Se incluyeron 112 pacientes con una edad media de 8,3 años. El fármaco más utilizado fue el propofol (64,3%), seguido de la asociación de ketamina con midazolam (16,1%) y del sevofluorano (12,5%). En el 70,5% de los pacientes se alcanzó un nivel de sedación profunda, sin diferencias estadísticamente significativas entre los distintos fármacos. Se registraron efectos adversos en un 51,8% de pacientes, principalmente desaturación, con una frecuencia mayor al emplear propofol (p< 0,05). La puntuación en la satisfacción alcanzó el valor máximo en todos los padres encuestados, sin hallarse diferencias significativas en función del procedimiento, fármaco, nivel de sedación o efectos adversos. En el 80% de los profesionales la puntuación alcanzó ese mismo valor. CONCLUSIONES: El fármaco más utilizado y con mayor eficacia es el propofol, aunque se asocia más frecuentemente con efectos adversos. El nivel de sedoanalgesia fue adecuado en el momento de iniciar los procedimientos. El grado de satisfacción es óptimo en la mayor parte de los encuestados, aunque fue registrado en menos de la mitad de los procedimientos

OBJECTIVES: To analyze the drugs most used for sedoanalgesia in procedures performed in a Pediatric Intensive Care Unit (PICU), to measure their effectiveness (level of sedation achieved), the main side effects and to carry out a quality control. MATERIAL AND METHODS: Prospective, observational and descriptive study. Epidemiological and clinical data, drug (s) used, level of sedation achieved, incidences or adverse effects, and satisfaction scales were collected from patients from 0 to 18 years of age who underwent procedures that required sedation and analgesia. RESULTS: 112 patients with an average age of 8.3 years were included. The most widely used drug was propofol (64.3%), followed by the association of ketamine with midazolam (16.1%) and sevofluorane (12.5%). In 70.5% of the patients, a level of deep sedation was reached, with no statistically significant differences between the different drugs used. Adverse effects were recorded in 51.8% of patients, mainly desaturation, with a higher frequency when using propofol (p <0.05). Satisfaction score was maximal in all the parents surveyed, without finding significant differences based on the procedure, drug, level of sedation or adverse effects. In 80% of the professionals the score was also maximal. CONCLUSION: The most used and with the highest efficacy in absolute values drug was propofol, although it was more frequently associated with adverse effects. The level of sedoanalgesia was adequate at the time of initiating the procedures. The degree of satisfaction was optimal in most of the respondents, although it was registered in less than half of the procedures

Microbial mucosal colonic shifts associated with the development of colorectal cancer reveal the presence of different bacterial and archaeal biomarkers.

BACKGROUND: Epidemiological studies demonstrate a link between gastrointestinal cancers and environmental factors such as diet. It has been suggested that environmental cancer risk is determined by the interaction between diet and microbes. Thus, the purpose of this study was to examine the hypothesis that microbiota composition during colorectal cancer (CRC) progression might differ depending on the stage of the disease. METHODS: A total of 28 age-matched and sex-matched subjects, seven with CRC adenocarcinoma, 11 with tubular adenomas and ten healthy subjects with intact colon, were included into the study. Microbiomes from mucosal and fecal samples were analyzed with 16S ribosomal RNA gene pyrosequencing, together with quantitative PCR of specific bacteria and archaea. RESULTS: The principal coordinates analysis clearly separated healthy tissue samples from polyps and tumors, supporting the presence of specific bacterial consortia that are associated with affected sites and that can serve as potential biomarkers of CRC progression. A higher presence of Fusobacterium nucleatum and Enterobacteriaceae was found by qPCR in samples from CRC compared to healthy controls. We observed a correlation between CRC process development and levels of Methanobacteriales (R = 0.537, P = 0.007) and Methanobrevibacterium (R = 0.574, P = 0.03) in fecal samples. CONCLUSION: Differences in microbial and archaeal composition between mucosal samples from healthy and disease tissues were observed in tubular adenoma and adenocarcinoma. In addition, microbiota from mucosal samples represented the underlying dysbiosis, whereas fecal samples seem not to be appropriate to detect shifts in microbial composition. CRC risk is influenced by microbial composition, showing differences according to disease progression step and tumor severity.

Molecular and functional characterization of LRP1 promoter polymorphism c.1-25 C>G (rs138854007).

The transcription of the Low-density lipoprotein receptor-related protein (LRP1) is upregulated by aggregated LDL (agLDL) and angiotensin II (AngII) in human vascular smooth muscle cells (hVSMC). The polymorphism c.1-25C>G creates a new GC-box in the LRP1 promoter recognized by Sp1/Sp3 transcription factors. The aims of this study were 1) to evaluate the impact of c.1-25C>G polymorphism on LRP1 transcriptional activity and expression, and 2) to examine the response of c.1-25C>G LRP1 promoter to LDL and AngII. EMSA and Luciferase assays in HeLa cells showed that -25G promoter has enhanced basal transcriptional activity and specific Sp1/Sp3 binding. hVSMC with GG genotype (GG-hVSMC) had higher LRP1 mRNA and protein levels, respectively than CC genotype (CC-hVSMC). EMSA assays showed that the polymorphism determines scarce amount of SRE-B/SREBP-2 complex formation and the failure of agLDL to further reduce these SRE-B/SREBP-2 complexes. Taken together, these results suggest that c.1-25C>G, by difficulting SREBP-2 binding, prevents SREBP-2 displacement required for LRP1 promoter response to LDL. In contrast, c.1-25C>G strongly favours Sp1/Sp3 binding and AngII-induced activity in Sp1/Sp3 dependent manner in GG-hVSMC. This increase is functionally translated into a higher capacity of GG-hVSMC to become foam cells from agLDL in presence of AngII. These results suggest that c.1-25C>G determines a lack of response to agLDL and an exacerbated response to AngII. It is thus conceivable that the presence of the polymorphism would be easily translated to vascular alterations in the presence of the pro-hypertensive autacoid, AngII.

Lipopolysaccharide downregulates CD91/low-density lipoprotein receptor-related protein 1 expression through SREBP-1 overexpression in human macrophages.

Sterol regulatory element-binding proteins (SREBPs) negatively modulate the expression of the CD91/low-density lipoprotein receptor-related protein (LRP1), a carrier and signaling receptor that mediates the endocytosis of more than 40 structurally and functionally distinct ligands. The aim of this work was to analyze whether lipopolysaccharide (LPS) can regulate LRP1 expression through SREBPs in human monocyte-derived macrophages (HMDM). LPS led to LRP1 mRNA and protein inhibition in a dose- and time-dependent manner. Concomitantly, a strong upregulation of SREBP-1 mRNA and SREBP-1 nuclear protein levels was observed in LPS-treated HMDM. The specific silencing of SREBP-1 efficiently prevented LRP1 reduction caused by LPS. SREBP-1 mRNA and nuclear protein levels remained high in HMDM treated with LPS unexposed or exposed to LDL. Native (nLDL) or aggregated LDL (agLDL) per se downregulated SREBP-2 expression levels and increased LRP1 expression. However, lipoproteins did not significantly alter the effect of LPS on SREBP-1 and LRP1 expression. Collectively, these data support that lipoproteins and LPS exert their modulatory effect on LRP1 expression through different SREBP isoforms, SREBP-2 and SREBP-1, respectively. These results highlight a crucial role of SREBP-1 as a mediator of the downregulatory effects of LPS on LRP1 expression in human macrophages, independently of the absence or presence of modified lipoproteins.

Selective role of sterol regulatory element binding protein isoforms in aggregated LDL-induced vascular low density lipoprotein receptor-related protein-1 expression.

Low density lipoprotein receptor-related protein (LRP1) is upregulated in vascular smooth muscle cells by intravascular aggregated LDL (agLDL) - LDL trapped in the arterial intima and systemic LDL. LRP1 upregulation in hypercholesterolemic aortas is concomitant with SREBP downregulation. However, the specific role of SREBP isoforms in LRP1 transcription and LDL-induced LRP1 upregulation in human vascular smooth muscle cells (VSMC) is unknown. In the present study we report that specific silencing of either SREBP-1 or SREBP-2 enhanced LRP1 whereas overexpression of the active SREBP isoforms decreased LRP1 expression. Gel mobility shift and ChIP assays demonstrated that SREBP-1a, SREBP-1c and SREBP-2 were able to bind to three putative SRE sequences; SRE-A (-1042 to -1028), SRE-B (-115 to -101) and SRE-C (+226 to +234). ChIP assays demonstrated that agLDL (100µg/mL, 24h) significantly and specifically decreased SREBP-2 binding to the LRP1 promoter. Luciferase assays demonstrated that agLDL increased the transcriptional activity of A/B or A/C double mutants but failed to increase that of the double B/C mutant. Our results show that both SREBP-1 and SREBP-2 negatively modulated LRP1 transcription. Furthermore, agLDL exerted an upregulatory effect on LRP1 expression by decreasing SREBP-2 binding to LRP1 promoter. Two SRE-like sequences control the response of LRP1 to agLDL.

Sterol regulatory element-binding protein-2 negatively regulates low density lipoprotein receptor-related protein transcription.

Low density lipoprotein receptor-related protein (LRP1) binds aggregated LDL (agLDL) leading to a high intracellular cholesteryl ester (CE) accumulation. AgLDL up-regulates LRP1 expression concomitantly with an LDL receptor (LDLR) and sterol regulatory element binding protein (SREBP-2) down-regulation. The objectives were to investigate whether SREBP-2 regulates LRP1 transcription and determine the molecular mechanisms involved in the process. Down-regulation of active SREBP-2 by nLDL and agLDL led to LDLR down-regulation and LRP1 up-regulation. Enforced expression of an active form of SREBP-2 (SREBP-2-NT, amino acid residues 1-468) decreased LRP1 expression and LRP1 promoter (WT-LRP1) luciferase activity in a dose-dependent manner. LDL did not exert any significant effect on LRP1 promoter activity when a putative sterol regulatory element (SRE) (5-GTGGGGTGA-3'; +225 to +233) was mutated (SRE-MT-LRP1). SREBP-2 overexpression exerted stronger down-regulatory effects on WT-LRP1 than on SRE-MT-LRP1 promoter activity both in control, nLDL- and agLDL-exposed HeLa cells. Gel mobility shift assays showed that recombinant SREBP-2-NT protein (1-468) binds to a double-stranded LRP1 DNA fragment (215 to 245) containing a wild-type (wt) SRE sequence but not to a mutated SRE (mt) sequence (5-GAATTCGA-3'). Our results demonstrate that LDL stimulates LRP1 transcription and decreases SREBP-2 active form which negatively regulates LRP1 transcription. SRE sequence (+225 to +233) plays a pivotal role for the down-regulatory effect of SREBP-2 on LRP1 promoter activity.