[Preparation and characterization of Ad-ERα-36-Fc-GFP].

ERα-36 is a novel subtype of estrogen receptor α which promotes tumor cell proliferation, invasion and drug resistance, and it serves as a therapeutic target. However, only small-molecule compounds targeting ERα-36 are under development as anticancer drugs at present. Gene therapy approach targeting ERα-36 can be explored using recombinant adenovirus armed with decoy receptor. The recombinant shuttle plasmid pDC316-Ig κ-ERα-36-Fc-GFP was constructed via genetic engineering to express an Ig κ-signaling peptide-leading secretory recombinant fusion protein ERα-36-Fc. The recombinant adenovirus Ad-ERα-36-Fc-GFP was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system. The expression of fusion protein and functional outcome of Ad-ERα-36-Fc-GFP transduction were further analyzed with triple-negative breast cancer MDA-MB-231 cells. Results showed that the recombinant adenovirus Ad-ERα-36-Fc-GFP was successfully generated. The virus effectively infected MDA-MB-231 cells which resulted in expression and secretion of the recombinant fusion protein ERα-36-Fc, leading to significant inhibition of EGFR/ERK signaling pathway. Preparation of the recombinant adenovirus Ad-ERα-36-Fc-GFP provides a basis for further investigation on cancer gene therapy targeting ERα-36.

AdeR-AdeS mutations & overexpression of the AdeABC efflux system in ciprofloxacin-resistant Acinetobacter baumannii clinical isolates.

Background & objectives: : Overexpression of efflux pumps is a cause of acquired resistance to fluoroquinolones in Acinetobacter baumannii. The present study was done to investigate the presence and overexpression of AdeABC efflux system and to analyze the sequences of AdeR-AdeS regulatory system in ciprofloxacin-resistant A. baumannii isolates. Methods: : Susceptibility of 50 clinical A. baumannii isolates to ciprofloxacin, imipenem, ceftazidime, cefepime and gentamicin antimicrobials was evaluated by agar dilution method. Isolates were screened for the evidence of active efflux pump. Isolates were also examined for adeR-adeS and adeB efflux genes by polymerase chain reaction (PCR). The adeR and adeS regulatory genes were sequenced to detect amino acid substitutions. Expression of adeB was evaluated by quantitative reverse-transcriptase PCR. Results: : There were high rates of resistance to ciprofloxacin (88%), ceftazidime (88%), cefepime (74%) and imipenem (72%) and less resistance rate to gentamicin (64%). Phenotypic assay showed involvement of active efflux in decreased susceptibility to ciprofloxacin among 16 isolates. The 12.27-fold increase and 4.25-fold increase were found in adeB expression in ciprofloxacin-full-resistant and ciprofloxacin-intermediate-resistant isolates, respectively. Several effective mutations, including A91V, A136V, L192R, A94V, G103D and G186V, were detected in some domains of AdeR-AdeS regulators in the overexpressed ciprofloxacin-resistant isolates. Interpretation & conclusions: The results of this study indicated that overexpression of the AdeABC efflux pump was important to reduce susceptibility to ciprofloxacin and cefepime in A. baumannii that, in turn, could be triggered by alterations in the AdeR-AdeS two-component system. However, gene expression alone does not seem adequate to explain multidrug resistance phenomenon. These results could help plan improved active efflux pump inhibitors.

Relationship between expression and antimicrobial resistance of RND efflux pump of Acinetobacter baumannii / 中国感染控制杂志

Objective To detect the distribution of resistance-nodulation (RND)efflux pump system of Acineto-bacter baumannii (AB),and explore the relationship between its’expression and antimicrobial resistance.Methods Fifty-nine strains of multidrug-resistant AB isolated from clinical specimens in The First Affiliated Hospital of Nan-chang University were identified and performed antimicrobial susceptibility analysis,distribution of RND efflux sys-tem of AB was detected by polymerase chain reaction(PCR),expression of efflux pump genes in different drug-re-sistant phenotypes of AB was compared,relationship between the expression level and drug resistance was analyzed, amplified products of RND efflux system were sequenced.Results Resistance rates of AB to ampicillin/sulbactam, imipenem,gentamicin,ciprofloxacin,and levofloxacin were 93.2%,94.9%,88.1%,96.6%,and 52.5% respec-tively.PCR detection results of efflux pump and integron genes of 59 AB strains revealed that the carrying rates of adeR,adeS,adeB,adeJ,and adeG genes were 81.4%,91.5%,93.2%,100.0%,and 61.0% respectively.The expression of efflux pump genes in different strains was different,expression levels of ade B and adeJ genes among gentamicin,imipenem,ampicillin/sulbactam resistant AB group and non-resistant AB group were significantly dif- ferent (all P＜0.05).There was no mutation or insertion sequence in the base sequences of regulatory genes ade R and ade S of adeABC efflux pump.Conclusion RND efflux pump system is universally present in AB,the expres-sion upregulation of ade B and ade J genes in RND efflux pump system is related with antimicrobial resistance of bacteria to gentamycin,imipenem,and ampicillin-sulbactam.

Higher-order accurate space-time schemes for computational astrophysics-Part I: finite volume methods.

As computational astrophysics comes under pressure to become a precision science, there is an increasing need to move to high accuracy schemes for computational astrophysics. The algorithmic needs of computational astrophysics are indeed very special. The methods need to be robust and preserve the positivity of density and pressure. Relativistic flows should remain sub-luminal. These requirements place additional pressures on a computational astrophysics code, which are usually not felt by a traditional fluid dynamics code. Hence the need for a specialized review. The focus here is on weighted essentially non-oscillatory (WENO) schemes, discontinuous Galerkin (DG) schemes and PNPM schemes. WENO schemes are higher order extensions of traditional second order finite volume schemes. At third order, they are most similar to piecewise parabolic method schemes, which are also included. DG schemes evolve all the moments of the solution, with the result that they are more accurate than WENO schemes. PNPM schemes occupy a compromise position between WENO and DG schemes. They evolve an Nth order spatial polynomial, while reconstructing higher order terms up to Mth order. As a result, the timestep can be larger. Time-dependent astrophysical codes need to be accurate in space and time with the result that the spatial and temporal accuracies must be matched. This is realized with the help of strong stability preserving Runge-Kutta schemes and ADER (Arbitrary DERivative in space and time) schemes, both of which are also described. The emphasis of this review is on computer-implementable ideas, not necessarily on the underlying theory.

AdeR protein regulates adeABC expression by binding to a direct-repeat motif in the intercistronic spacer.

Overexpression of the efflux pump AdeABC is associated with tigecycline resistance of multi-drug resistant Acinetobacter baumannii (MDRAB). A two-component regulatory system, sensor AdeS and regulator AdeR proteins regulate the pump. However, the detailed mechanism of the AdeR protein to enhance the expression of adeABC operon is not well defined. We illustrated the biological characteristics of AdeR proteins by comparing a mutant AdeR protein of a tigecycline resistant MDRAB to the wild AdeR protein. By analyzing a series of deletion constructs, a minimal gene cassette of the intercistronic spacer DNA fragment specifically bound with the adeR protein and resulted in band shifting in electrophoresis mobility shifting assays (EMSA). A conserve direct repeat motif was observed in the intercistronic spacer DNA. We demonstrated the AdeR protein was a direct-repeat-binding protein. Two common residue mutations on the AdeR proteins of tigecycline resistant MDRAB isolates could reduce their binding affinity with the intercistronic spacer. The free intercistronic spacer may then more efficiently support the read-through of the adeABC operon during the co-transcriptional translation in tigecycline resistant MDRAB isolates.

Efficient conservative ADER schemes based on WENO reconstruction and space-time predictor in primitive variables.

We present a new version of conservative ADER-WENO finite volume schemes, in which both the high order spatial reconstruction as well as the time evolution of the reconstruction polynomials in the local space-time predictor stage are performed in primitive variables, rather than in conserved ones. To obtain a conservative method, the underlying finite volume scheme is still written in terms of the cell averages of the conserved quantities. Therefore, our new approach performs the spatial WENO reconstruction twice: the first WENO reconstruction is carried out on the known cell averages of the conservative variables. The WENO polynomials are then used at the cell centers to compute point values of the conserved variables, which are subsequently converted into point values of the primitive variables. This is the only place where the conversion from conservative to primitive variables is needed in the new scheme. Then, a second WENO reconstruction is performed on the point values of the primitive variables to obtain piecewise high order reconstruction polynomials of the primitive variables. The reconstruction polynomials are subsequently evolved in time with a novel space-time finite element predictor that is directly applied to the governing PDE written in primitive form. The resulting space-time polynomials of the primitive variables can then be directly used as input for the numerical fluxes at the cell boundaries in the underlying conservative finite volume scheme. Hence, the number of necessary conversions from the conserved to the primitive variables is reduced to just one single conversion at each cell center. We have verified the validity of the new approach over a wide range of hyperbolic systems, including the classical Euler equations of gas dynamics, the special relativistic hydrodynamics (RHD) and ideal magnetohydrodynamics (RMHD) equations, as well as the Baer-Nunziato model for compressible two-phase flows. In all cases we have noticed that the new ADER schemes provide less oscillatory solutions when compared to ADER finite volume schemes based on the reconstruction in conserved variables, especially for the RMHD and the Baer-Nunziato equations. For the RHD and RMHD equations, the overall accuracy is improved and the CPU time is reduced by about 25 %. Because of its increased accuracy and due to the reduced computational cost, we recommend to use this version of ADER as the standard one in the relativistic framework. At the end of the paper, the new approach has also been extended to ADER-DG schemes on space-time adaptive grids (AMR).

Factores asociados con la adherencia a la actividad física en el tiempo libre / Fatores associados com a aderência à atividade física no tempo libre / Associated factors with adherence to physical activity in leisure time

Introducci¢n: existe en Colombia poco conocimiento respecto a los facto- res asociados con la adherencia al ejercicio. Objetivo: evaluar los factores asociados con la adherencia a la actividad f¡sica en el tiempo libre (AFTL) de los adultos que participan en un programa de actividad f¡sica (AF). M‚- todos: se realiz¢ un estudio prospectivo de Casos y Controles. Se defini¢ como caso a quien realiz¢ durante un periodo de 5 meses un promedio 150 minutos o m s de AFTL por semana a una intensidad moderada ¢ 75 minutos a una intensidad vigorosa; como control se tom¢ a quien no cum- pli¢ con los anteriores requisitos. Se evaluaron variables independientes sociodemogr ficas, factores de riesgo cardiovascular y osteomuscular, y las motivacionales para la pr ctica del ejercicio. Los datos se analizaron por medio de un modelo de regresi¢n log¡stica multivariado. Resultados: la muestra la conformaron 176 personas usuarias de un programa de AF, en su mayor¡a mujeres (69,3%); el promedio de edad fue de 55,7 a¤os. Las variables asociadas con la adherencia a la AFTL fueron el antecedente personal de artrosis (OR 0,392; IC95% 0,184-0,837; p=0,016) y la edad (OR 1,027; IC95% 1,001-1,054; p=0,041), las cuales explicaron el 8,0% de la variabilidad. Conclusiones: las variables que se asociaron en forma independiente con la adherencia a la AFTL fueron la edad y el anteceden- te personal de artrosis. Se requiere realizar nuevos estudios que incluyan otras variables que puedan explicar en forma m s global este fen¢meno.

Introdu‡Æo: Existe na Col“mbia pouco conhecimento a respeito dos fatores associados com a aderˆncia ao exerc¡cio. Objetivo: Avaliar os fatores associados com a aderˆncia … atividade f¡sica no tempo livre (AFTL) dos adultos que participam em um programa de atividade f¡sica (AF). M‚todos: Se realizou um estudo prospectivo de casos e controles. Definiu-se como caso a quem realizou durante um per¡odo de 5 meses em m‚dia 150 minutos ou mais de AFTL por semana, a uma intensidade moderada ou 75 minutos a uma intensidade vigorosa; como controle tomou-se a quem nÆo cumpriu com os requisitos anteriores. Avaliou-se vari veis independentes s¢cios demogr ficas, fatores de risco cardiovascular, osteomuscular e as motiva‡ães para a pr tica de exerc¡cios. Analisaram-se os dados por meio de um modelo de regressÆo log¡stica multivariado. Resultados: conformaram a amostra 176 pessoas usu rias de um programa de AF, em sua maioria mulheres (69,3%); a m‚dia de idade foi de 55,7 anos. As vari veis associadas com a aderˆncia a AFTL foram o antecedente pessoal de artrose (OR 0,392; IC95% 0,184-0,837; p=0,016) e a idade (OR 1,027; IC95% 1,001-1,054; p=0,041), os quais explicaram os 8% de variabilidade. Conclusães: As vari veis que se associaram de forma independente com a aderˆncia … AFTL foram a idade e o antecedente pessoal de artrose. Faz-se necess rio realizar novos estudos que incluam outras vari veis que possam explicar de forma mais global este fen“meno.

Introduction: In Colombia there is a low knowledge related to the associated factors with adherence to exercise. Objective: To evaluate the associated factors with adherence to the physical activity in leisure time in adults (PALT) who participate in a physical activity program. Methods: The study was a prospective case-control. A case was defined as who performed during five months an average of 150 minutes or more per week of PALT, with a moderate intensity or strong intensity of 75 minutes or more of PALT per week; as a control was had in consideration, people who did not meet the above criteria. The independent variables measured were: socio-demographic factors, cardiovascular, musculoskeletal risk factors, and the motivation to exercise. The data was analyzed through a varied regressive logistic model. Results: The sample involved 176 female users of a PA program (69.3%); with an average age of 55.7 years old. The associated variables with the PALT adherence were the personal osteoarthritis record (OR 0,392; IC95% 0,184 – 0,837; P=0,016) and the age (OR 1,027; IC95% 1,001-1,054; p=0,041), which explained 8, 0 % of the variability. Conclusions: The variables that were independently associated to the adherence to PALT were age, and the personal osteoarthritis record. There is a necessity of new studies that include new variables to allow a global analysis of the phenomena.

A global multiscale mathematical model for the human circulation with emphasis on the venous system.

We present a global, closed-loop, multiscale mathematical model for the human circulation including the arterial system, the venous system, the heart, the pulmonary circulation and the microcirculation. A distinctive feature of our model is the detailed description of the venous system, particularly for intracranial and extracranial veins. Medium to large vessels are described by one-dimensional hyperbolic systems while the rest of the components are described by zero-dimensional models represented by differential-algebraic equations. Robust, high-order accurate numerical methodology is implemented for solving the hyperbolic equations, which are adopted from a recent reformulation that includes variable material properties. Because of the large intersubject variability of the venous system, we perform a patient-specific characterization of major veins of the head and neck using MRI data. Computational results are carefully validated using published data for the arterial system and most regions of the venous system. For head and neck veins, validation is carried out through a detailed comparison of simulation results against patient-specific phase-contrast MRI flow quantification data. A merit of our model is its global, closed-loop character; the imposition of highly artificial boundary conditions is avoided. Applications in mind include a vast range of medical conditions. Of particular interest is the study of some neurodegenerative diseases, whose venous haemodynamic connection has recently been identified by medical researchers.

Well-balanced high-order solver for blood flow in networks of vessels with variable properties.

We present a well-balanced, high-order non-linear numerical scheme for solving a hyperbolic system that models one-dimensional flow in blood vessels with variable mechanical and geometrical properties along their length. Using a suitable set of test problems with exact solution, we rigorously assess the performance of the scheme. In particular, we assess the well-balanced property and the effective order of accuracy through an empirical convergence rate study. Schemes of up to fifth order of accuracy in both space and time are implemented and assessed. The numerical methodology is then extended to realistic networks of elastic vessels and is validated against published state-of-the-art numerical solutions and experimental measurements. It is envisaged that the present scheme will constitute the building block for a closed, global model for the human circulation system involving arteries, veins, capillaries and cerebrospinal fluid.