Controlling instabilities along a 3DVar analysis cycle by assimilating in the unstable subspace: a comparison with the EnKF

A hybrid scheme obtained by combining 3DVar with the Assimilation in the Unstable Subspace (3DVar-AUS) is tested in a QG model, under perfect model conditions, with a fixed observational network, with and without observational noise. The AUS scheme, originally formulated to assimilate adaptive observations, is used here to assimilate the fixed observations that are found in the region of local maxima of BDAS vectors (Bred vectors subject to assimilation), while the remaining observations are assimilated by 3DVar. The performance of the hybrid scheme is compared with that of 3DVar and of an EnKF. The improvement gained by 3DVar-AUS and the EnKF with respect to 3DVar alone is similar in the present model and observational configuration, while 3DVar-AUS outperforms the EnKF during the forecast stage. The 3DVar-AUS algorithm is easy to implement and the results obtained in the idealized conditions of this study encourage further investigation toward an implementation in more realistic contexts

Identification of domains in apoA-I susceptible to proteolysis by mast cell chymase. Implications for HDL function.

When stimulated, rat serosal mast cells degranulate and secrete a cytoplasmic neutral protease, chymase. We studied the fragmentation of apolipoprotein (apo) A-I during proteolysis of HDL(3) by chymase, and examined how chymase-dependent proteolysis interfered with the binding of eight murine monoclonal antibodies (Mabs) against functional domains of apoA-I. Size exclusion chromatography of HDL(3) revealed that proteolysis for up to 24 h did not alter the integrity of the alpha-migrating HDL, whereas a minor peak containing particles of smaller size with prebeta mobility disappeared after as little as 15 min of incubation. At the same time, generation of a large (26 kDa) polypeptide containing the N-terminus of apoA-I was detected. This large fragment and other medium-sized fragments of apoA-I produced after prolonged treatment with chymase were found to be associated with the alphaHDL; meanwhile, small lipid-free peptides were rapidly produced. Incubation of HDL(3) with chymase inhibited binding of Mab A-I-9 (specific for prebeta(1)HDL) most rapidly (within 15 min) of the eight studied Mabs. This rapid loss of binding was paralleled by a similar reduction in the ability of HDL(3) to induce high-affinity efflux of cholesterol from macrophage foam cells, indicating that proteolysis had destroyed an epitope that is critical for this function. In sharp contrast, prolonged degradation of HDL(3) by chymase failed to reduce the ability of HDL(3) to activate LCAT, even though it led to modification of three epitopes in the central region of apoA-I that are involved in lecithin cholesterol acyltransferase (LCAT) activation. This differential sensitivity of the two key functions of HDL(3) to the proteolytic action of mast cell chymase is compatible with the notion that, in reverse cholesterol transport, intactness of apoA-I is essential for prebeta(1)HDL to promote the high-affinity efflux of cellular cholesterol, but not for the alpha-migrating HDL particles to activate LCAT

Comparative mapping of the fragile histidine triad (FHIT) gene in cattle, river buffalo, sheep and goat by FISH and assignment to BTA22 by RH-mapping: a comparison with HSA3

Common fragile sites can be damaged by exposure to a variety of carcinogens. The fragile histidine triad (FHIT) gene, including the most active human chromosomal fragile site (FRA3B) at chromosome band HSA3p14.2,1 has been proposed as a tumour suppressor gene for a variety of tumours.2 The most common response to carcinogen exposure is deletions at the FHIT locus that alter the gene structure and function. In this study we assign the FHIT gene in cattle, river buffalo, sheep and goat chromosomes by comparative fluorescence in situ hybridization (FISH)-mapping. In addition, the assignment to BTA22 was confirmed by typing the marker across a bovine radiation hybrid (RH) panel

Cost of Bordetella pertussis illness in tertiary hospitals in Argentina

La Comisión Nacional de Inmunizaciones y el ProNaCEI (Programa Nacional de Control de Enfermedades Inmunoprevenibles) actualizaron la política de vacunación por Bordetella pertussis (BP) a partir del año 2009 con el objetivo de optimizar el control de esta enfermedad, de acuerdo con las recomendaciones internacionales. Para evaluar el impacto económico de esta nueva política de vacunación resulta necesario conocer inicialmente el costo que implica para el sistema de salud un niño internado o ambulatorio con infección por BP. El objetivo de este estudio fue describir el perfl de costos en niños internados o tratados ambulatoriamente, con infección confrmada por laboratorio de BP en tres hospitales de la Argentina. Estudio prospectivo de costo de la enfermedad durante el período diciembre de 2010 a marzo de 2012. Resultados. El costo total para toda la cohorte fue de 1 170 663,32 pesos (236 497,64 dólares); los costos médicos directos, de 1 124 052,31 pesos (227 081,27 dólares); los costos indirectos y gastos de bolsillo, de 46611 pesos (9 416,36 dólares), lo que permite inferir un costo total promedio por paciente de 10 546,52 pesos (IC 95% 9009 a 13 840) (2130,60 dólares, IC 95% 1820 a 2795), costos médicos directos por paciente de 10 126,6 pesos (IC 95% 8607 a 13 171) (2045,77 dólares, IC 95%1738 a 2660) y costos indirectos más de bolsillo (viajes y extras) de 419,92 pesos (IC 95% 344,7 a 565,3), (84 dólares, IC 95% 69 a 115). Conclusión. El costo de un caso confrmado hospitalizado por BP es 10 546,52 pesos (IC 95% 9009 a 13 840) (2130,60 dólares, IC 95% 1820 a 2795). Los costos directos no médicos y costos indirectos constituyen el 4% del total, lo que corresponde a 419,91 pesos por familia (84 dólares, IC 95% 69 a 115), un 8% del salario promedio.Fil: Gentile, Angela. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "ricardo Gutierrez"; ArgentinaFil: Salgueiro, Ana L.. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "ricardo Gutierrez"; ArgentinaFil: Garcia Bournissen, Facundo. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "r. Gutierrez". Servicio de Parasitología y Chagas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romanin, Viviana. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "ricardo Gutierrez"; ArgentinaFil: Bulgheroni, Sonia. Hospital Materno Infantil de San Isidro "Dr. Gianantonio"; ArgentinaFil: Gaiano, Alejandra. Hospital Materno Infantil de San Isidro "Dr. Gianantonio"; ArgentinaFil: Benegas, Liliana. Hospital de Niños "Víctor J. Vilela"; ArgentinaFil: Uboldi, Andrea. Hospital de Niños "Víctor J. Vilela"; ArgentinaFil: Giglio, Norberto. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "ricardo Gutierrez"; Argentin

Monoclonal antibodies to human low density lipoprotein identify distinct areas on apolipoprotein B-100 relevant to the low density lipoprotein-receptor interaction.

We have characterized the epitopes for ten murine monoclonal antibodies (Mabs) to human low density lipoprotein (LDL) and studied their ability to interfere with the LDL-receptor interaction. The epitopes for the antibodies were defined by using the following approaches: 1) interaction with apoB-48; 2) interaction with apoB-100 thrombolytic fragments; and 3) interaction with beta-galactosidase-apoB fusion proteins spanning different areas of the apoB-100 sequence. The results obtained are consistent with the following map of epitopes: Mab 6E, amino acids (aa) 1-1297, Mabs 5A and 6B, aa 1480-1693, Mabs 2A, 7A, 3B, and 4B, aa 2152-2377, Mabs 8A and 9A, aa 2657-3248 and 3H, aa 4082-4306. Four Mabs (2A, 5A, 7A, and 9A) whose epitopes are located in three different areas of apoB, dramatically reduced (up to 95%) the LDL-receptor interaction on cultured human fibroblasts; Fab fragments were as effective as the whole antibodies. Mab 3H, on the other hand, increased LDL binding up to threefold. These findings are consistent with the hypothesis that several areas of apoB-100 are involved independently or in concert in modulating the apoprotein B conformation required for interaction with the LDL receptor

Predictive value of HDL function in patients with coronary artery disease: relationship with coronary plaque characteristics and clinical events

BACKGROUND: HDL is endowed with several metabolic, vascular, and immunoinflammatory protective functions. Among them, a key property is to promote reverse cholesterol transport from cells back to the liver. The aim of this study was to estimate the association of scavenger receptor class B type I (SR-BI)- and ATP binding cassette transporter A1 (ABCA1)-mediated cholesterol efflux (the two major routes for cholesterol efflux to HDL) with the presence, extent, and severity of coronary artery disease (CAD), vascular wall remodelling processes, coronary plaque characteristics, and the incidence of myocardial infarction in the different subgroups of patients from the CAPIRE study. METHODS: Patients (n = 525) from the CAPIRE study were divided into two groups: low-risk factors (RF), with 0–1 RF (n = 263), and multiple-RF, with ≥2 RFs; within each group, subjects were classified as no-CAD or CAD based on the segment involvement score (SIS) evaluated by coronary computed tomography angiography (SIS = 0 and SIS > 5, respectively). SR-BI- and ABCA1-mediated cholesterol efflux were measured using the plasma of all patients. RESULTS: SR-BI-mediated cholesterol efflux was significantly reduced in patients with CAD in both the low-RF and multiple-RF groups, whereas ABCA1-mediated cholesterol efflux was similar among all groups. In CAD patients, multivariable analysis showed that SR-BI-mediated cholesterol efflux <25(th) percentile predicted cardiovascular outcome (odds ratio 4.1; 95% CI: 1.3–13.7; p = .019), whereas ABCA-1-mediated cholesterol efflux and HDL-C levels significantly did not. Despite this finding, reduced SR-BI-mediated cholesterol efflux was not associated with changes in high-risk plaque features or changes in the prevalence of elevated total, non-calcified, and low-attenuation plaque volume. CONCLUSION: KEY MESSAGES: Increased cholesterol efflux capacity, an estimate of HDL function, is associated with a reduced CVD risk, regardless of HDL-C levels. HDL-C levels are significantly lower in patients with CAD. Lower SR-BI-mediated cholesterol efflux capacity is observed in patients with diffuse coronary atherosclerosis and is associated with the worst clinical outcomes in patients with CAD, independently of atherosclerotic plaque features

Effect of Polyethylene Glycol Content and Molar Mass on Injection Molding of Hydroxypropyl Methylcellulose Acetate Succinate-Based Gastroresistant Capsular Devices for Oral Drug Delivery

Capsular devices for oral drug delivery were recently proposed and manufactured by injection molding (IM) as an evolution of traditional reservoir systems comprising a core and a functional coating. IM allowed the fabrication of capsule shells with release-controlling features based on the employed materials and the design characteristics. These features are independent of the drug, with significant savings in development time and costs. In previous work, IM was used to produce enteric-soluble capsules from blends of hydroxypropyl methylcellulose acetate succinate, with polyethylene glycol (PEG) as the plasticizer. In this work, the range of plasticizer concentrations and molar mass was broadened to evaluate in-depth how those parameters affect material processability and capsule performance over time. As expected, increasing the amount of the low molar mass plasticizer decreased the viscosity and modulus of the material. This simplified the molding process and enhanced the mechanical resistance of the shell, as observed during assembly. However, some samples turned out translucent, depending on several factors including storage conditions. This was attributed to plasticizer migration issues. Such results indicate that higher molar mass PEGs, while not significantly impacting on processability, lead to capsular devices with consistent performance in the investigated time lapse

PCSK9 deficiency reduces insulin secretion and promotes glucose intolerance: the role of the low-density lipoprotein receptor

Aims PCSK9 loss of function genetic variants are associated with lower low-density lipoprotein cholesterol but also with higher plasma glucose levels and increased risk of Type 2 diabetes mellitus. Here, we investigated the molecular mechanisms underlying this association. Methods and results Pcsk9 KO, WT, Pcsk9/Ldlr double KO (DKO), Ldlr KO, albumin AlbCre+/Pcsk9LoxP/LoxP (liver-selective Pcsk9 knock-out mice), and AlbCre-/Pcsk9LoxP/LoxP mice were used. GTT, ITT, insulin and C-peptide plasma levels, pancreas morphology, and cholesterol accumulation in pancreatic islets were studied in the different animal models. Glucose clearance was significantly impaired in Pcsk9 KO mice fed with a standard or a high-fat diet for 20\u2009weeks compared with WT animals; insulin sensitivity, however, was not affected. A detailed analysis of pancreas morphology of Pcsk9 KO mice vs. controls revealed larger islets with increased accumulation of cholesteryl esters, paralleled by increased insulin intracellular levels and decreased plasma insulin, and C-peptide levels. This phenotype was completely reverted in Pcsk9/Ldlr DKO mice implying the low-density lipoprotein receptor (LDLR) as the proprotein convertase subtilisin/kexin Type 9 (PCSK9) target responsible for the phenotype observed. Further studies in albumin AlbCre+/Pcsk9LoxP/LoxP mice, which lack detectable circulating PCSK9, also showed a complete recovery of the phenotype, thus indicating that circulating, liver-derived PCSK9, the principal target of monoclonal antibodies, does not impact beta-cell function and insulin secretion. Conclusion PCSK9 critically controls LDLR expression in pancreas perhaps contributing to the maintenance of a proper physiological balance to limit cholesterol overload in beta cells. This effect is independent of circulating PCSK9 and is probably related to locally produced PCSK9