bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic manipulation of myoblasts and a novel primary myosatellite cell culture system: comparing and optimizing approaches
Authors: Jackson Melissa F. , Hoversten Knut E. , Powers John M. , Trobridge Grant D. , Rodgers Buel D. 
Published: November 2012

The genetic manipulation of skeletal muscle cells in vitro is notoriously difficult, especially when using undifferentiated muscle cell lines (myoblasts) or primary muscle stem cells (myosatellites). We therefore optimized methods of gene transfer by overexpressing green fluorescent protein (GFP) in mouse C2C12 cells and in a novel system, primary rainbow trout myosatellite cells. A common lipid‐based transfection reagent was used (Lipofectamine 2000) along with three different viral vectors: adeno‐associated virus serotype 2 (AAV2), baculovirus (BAC) and lentivirus. Maximal transfection efficiencies of 49% were obtained in C2C12 cells after optimizing cell density and reagent : DNA ratio, although the GFP signal rapidly dissipated with proliferation and was not maintained with differentiation. The transduction efficiency of AAV2 was optimized to 65% by extending incubation time and decreasing cell density, although only 30% of cells retained expression after passing. A viral comparison revealed that lentivirus was most efficient at transducing C2C12 myoblasts as 97% of cells were transduced with only 106 viral genomes (vg) compared to 54% with 108 vg AAV2 and 23% with 109 vg BAC. Lentivirus also transduced 90% of primary trout myosatellites compared to 1–10% with AAV2 and BAC. The phosphoglycerate kinase 1 (pgk) promoter was 10‐fold more active than the cytomegalovirus immediate‐early promoter in C2C12 cells and both were effective in trout myosatellites. Maximal transduction of C2C12 myotubes was achieved by differentiating myoblasts previously transduced with lentivirus and the pgk promoter. Thus, our optimized protocol proved highly effective in diverse muscle cell systems and could therefore help overcome a common technological barrier.
Role of transcription factor Sp1 and RNA binding protein HuR in the downregulation of Dr+Escherichia coli receptor protein decay accelerating factor (DAF or CD55) by nitric oxide
Authors: Banadakoppa Manu , Liebenthal Daniel , Nowak David E. , Urvil Petri , Yallampalli Uma , Wilson Gerald M. , Kishor Aparna , Yallampalli Chandra 
Published: November 2012

We previously reported that nitric oxide (NO) reduces the rate of bacteremia and maternal mortality in pregnant rats with uterine infection by Escherichia coli expressing the Dr Fimbria (Dr+). The epithelial invasion of Dr+ E. coli is dependent on the expression level of its cellular receptor decay accelerating factor (DAF). NO reduces the rate of bacteremia by downregulating the expression of DAF. In this study, we elucidated the role of transcription factor Sp1 and RNA binding protein HuR in the downregulation of human DAF by NO. We generated a series of deletion mutant constructs of DAF gene 5′‐untranslated region and mapped the NO‐response region upstream to the core promoter region of the DAF gene. One of the several Sp1 binding sites in the DAF 5′‐untranslated region was located within the NO‐response region. The binding of Sp1 to this site was inhibited by NO. Furthermore, NO also promoted the degradation of DAF mRNA. The 3′‐untranslated region of DAF harbors an AU‐rich element and this element destabilized the mRNA transcript. NO promoted the rapid degradation of DAF mRNA by inhibiting the binding of mRNA stabilizing protein HuR to this AU‐rich region. The inhibition of binding of HuR to the AU‐rich region was due to the S‐nitrosylation of one or more cysteine residues by NO. Thus, these data reveal the molecular mediators of transcriptional and post‐transcriptional regulation of DAF by NO with implications in pathophysiology related to DAF.
Numerical simulation of the coupling between thermal dissipation and fish‐eye crack growth in very high cycle fatigue regime
Authors: NGUYEN H.Q. , GALLIMARD L. , BATHIAS C. 
Published: November 2012

ABSTRACTIn this paper, we study the temperature field associated with the propagation of a fatigue crack in a very high cycle fatigue regime during ultrasonic fatigue testing. We use a Paris–Hertzberg crack growth law to compute the evolution of the crack and a perfectly elastic–plastic constitutive law to compute the plastic dissipation per cycle at the tip of the crack. A thermomechanical finite element model is proposed to estimate the evolution of the temperature field during the crack propagation. Numerical results obtained agree fairly well with experimental results.
Assessment of the thrombin generation assay in haemophilia: Comparative study between fresh and frozen platelet‐rich plasma
Authors: Vila V. , Aznar J. A. , Moret A. , Marco A. , Navarro S. , Vila C. , España F. 
Published: November 2012

SummaryThe severity of haemophilia A has traditionally been classified by the dosage of factor VIII (FVIII) by one‐step coagulation tests. However, an homogeneous group of patients with similar FVIII levels show clinical heterogeneity and 10–15% of the patients classified as severe haemophilia do not have a severe bleeding phenotype. Traditional tests used for measuring FVIII are not capable of detecting other prohaemorrhagic or prothrombotic factors. Global tests as the thrombin generation assay (TGA) may detect these haemostatic factors. So TGA may be an additional tool for classifying the actual severity of haemophilia. Our group is carrying out correlation tests between FVIII and TGA in platelet‐poor and ‐rich plasmas (PPP and PRP, respectively). PRP has the inconvenience that must be done freshly soon after blood extraction. Our aim is to study the differences between TGA performed with fresh and frozen PRP and PPP and its implementation in multicenter studies. We included 70 patients with severe haemophilia A in prophylactic treatment. Venous blood drawing was obtained prior to administration of FVIII, at the trough levels. FVIII measurement and TGA were performed in fresh and frozen PRP and PPP. The platelet absence caused a significant decrease in TGA although PPP and PRP correlated well. Frozen samples gave different results in PPP, but there were no significant differences between fresh and frozen PRP. This fact enables using frozen PRP in multicenter studies with a TGA‐specialized laboratory for reclassifying haemophilia severity and for pharmacokinetic studies with TGA.
Management of surgical procedures in children with severe FV deficiency: experience of 13 surgeries
Authors: Mathias M. , Tunstall O. , Khair K. , Liesner R. 
Published: November 2012

SummaryHomozygous severe factor V (FV) deficiency has a prevalence of around one per million. Even in patients with FV levels of <0.01 IU mL−1 there appears to be a variation in bleeding phenotype in that there is a subgroup of affected individuals who present in later childhood and have a relatively mild bleeding phenotype, but there are children who present as neonates with intracerebral bleeding events and who have a much more severe bleeding phenotype. The only available current FV replacement is in the form of fresh frozen plasma (FFP) or solvent detergent FFP. We present here our experience with surgical haemostatic cover for 13 surgeries in three children with severe FV deficiency.
Correlation of connective tissue growth factor with liver stiffness measured by transient elastography in biliary atresia
Authors: Honsawek Sittisak , Udomsinprasert Wanvisa , Chirathaworn Chintana , Anomasiri Wilai , Vejchapipat Paisarn , Poovorawan Yong 
Published: November 2012

AimBiliary atresia (BA) is a neonatal liver disease defined as chronic progressive fibrotic obliteration of extrahepatic bile ducts. The objective of this study was to determine the association of serum connective tissue growth factor (CTGF) with clinical outcome and liver stiffness measurement.MethodsEighty‐two BA patients post‐Kasai operation and 28 healthy controls were recruited. BA patients were categorized into two groups based on their portal hypertension (PH) status. Serum CTGF levels were determined by enzyme‐linked immunosorbent assay. Liver stiffness scores were measured by transient elastography.ResultsBA patients had greater CTGF levels (905.9 ± 57.7 vs 238.3 ± 23.5 pg/mL, P < 0.001) and higher liver stiffness values than controls (28.2 ± 2.6 vs 5.0 ± 0.5 kPa, P < 0.001). Serum CTGF levels were remarkably elevated in BA patients with PH compared to those without PH (1092.4 ± 73.9 vs 582.6 ± 45.7 pg/mL, P < 0.001). Furthermore, BA patients with PH had significantly higher liver stiffness values compared to those without PH (37.3 ± 3.0 vs 10.6 ± 1.1 kPa, P < 0.001). Additionally, serum CTGF was positively correlated with liver stiffness (r = 0.875, P < 0.001) and total bilirubin (r = 0.462, P < 0.001). There was an inverse correlation between serum CTGF and serum albumin (r = −0.579, P < 0.001).ConclusionHigh serum CTGF was associated with a poor outcome in BA patients. Accordingly, serum CTGF and transient elastography may serve as non‐invasive biomarkers reflecting the disease severity in postoperative BA patients.
An evaluation of a Books on Prescription scheme in a UK public library authority
Authors: Furness Rebecca , Casselden Biddy 
Published: November 2012

AbstractThis article discusses an evaluation of a Books on Prescription (BOP) scheme in a UK public library authority. The research was carried out by Rebecca Furness and submitted as a dissertation for the MSc Information and Library Management to Northumbria University. The dissertation was supervised by Biddy Casselden at Northumbria University and was awarded a distinction. The dissertation identified areas for development for BOP schemes and made specific recommendations that could make the schemes more accessible, enabling significant numbers of people to lead more fulfilling lives. Because this study focuses on mental health and the role that UK public libraries have in supporting well‐being, it is a good illustration of the wide‐ranging nature of subjects welcomed for the Dissertations into practice feature.AM
Preparation and characteristics of squid pen β‐chitin prepared under optimal deproteinisation and demineralisation condition
Authors: Youn Dal Kyoung , No Hong Kyoon , Prinyawiwatkul Witoon 
Published: November 2012

SummarySquid (Todarodes pacifica) pen was an excellent source of β‐chitin with 25.5% yield. The optimal condition to prepare squid pen β‐chitin was established: deproteinisation with 3% NaOH for 30 min at 15 psi/121 °C and a solid/solvent ratio of 1:10 (w/v) and a subsequent demineralisation with 1 N HCl for 30 min at room temperature and a solid/solvent ratio of 1:10 (w/v). Squid pen β‐chitin contained 6.29% nitrogen, 0.25% ash, and negligible fat with degree of acetylation of 94.02%, residual amino acid of 0.499 g/100 g and bulk density of 0.28 g mL−1. Depending on its particle size, squid pen β‐chitin visually looked white (L* = 82.82, a* = −0.67, b* = 6.31; particle size of 0.15–0.18 mm) or light grey (L* = 62.88, a* = 0.33, b* = 10.66; particle size of 0.425–0.841 mm). Water, fat and dye‐binding capacity of squid pen β‐chitin was 694.67%, 194.03% and 79.81%, respectively.
Misapplication of the Clausius–Clapeyron equation for estimating heats of adsorption in foods
Authors: Caurie Matthew 
Published: November 2012

SummaryIt has been pointed out that the Clausius–Clapeyron equation which is currently used to measure isosteric heat of adsorption in reality measures the latent heat of evaporation. The usual Clausius–Clapeyron plot that decays rapidly to low values at low Aws has been compared with a suggested alternative equation derived directly from an expanded form of Caurie's unimolecular adsorption equation. The plot of the proposed equation is higher than that of the Clausius–Clapeyron equation, and it has the shape of an inverse sigmoid isotherm. Two discontinuities in the proposed plots are stated to demarcate three energy levels corresponding to three states of bound water recognised to exist along sigmoid food isotherm plots.
Watermelon juice pretreatment with microfiltration process for obtaining lycopene
Authors: Gomes Flávia S. , Costa Priscila A. , Campos Maria B. D. , Tonon Renata V. , Couri Sônia , Cabral Lourdes M. C. 
Published: November 2012

SummaryThe aim of this study was to evaluate the effect of microfiltration on the lycopene content and antioxidant capacity of watermelon juice using ceramic membranes with an average pore size of 0.1 μm and a permeable area of 0.022 m2. The influence of the tangential cross‐flow velocity (4.3–6.4 m s−1) and temperature (23–37 °C) on the permeation flux, lycopene content and antioxidant capacity was evaluated. A 22complete factorial experimental design was used. Increases in tangential cross‐flow velocity and temperature had positive effects on permeation flux, with a maximum of 92.4 l h−1 m−2achieved at 6.4 m s−1and 30 °C. With regard to the lycopene content and antioxidant capacity, tangential cross‐flow velocity was the only factor that exhibited significant effects on these responses. For the most concentrated extract, the lycopene concentration and antioxidant capacity were enhanced by 402.8% and 416.3%, respectively.
Acid gelation of native and heat‐denatured soy proteins and locust bean gum
Authors: Perrechil Fabiana A. , Braga Ana Luiza M. , Cunha Rosiane L. 
Published: November 2012

SummaryThe effects of protein concentration and locust bean gum (LBG) addition on the mechanical properties, microstructure and water holding capacity of acidified soy protein (SPI) gels were studied. The protein was employed in two different states: (i) native and (ii) heat denatured. A slow acidification rate was induced in both systems by applying glucono‐δ‐lactone (GDL). The results indicated that the gels of native SPI were weaker, less deformable and showed lower water holding capacity than the gels of heat‐denatured SPI. The LBG addition led to an increase in the strength and water holding capacity of SPI gels, independent of the protein state (native or denatured). These results indicated that the properties of texture and water holding capacity of the SPI acid gels can be modulated by the process conditions or by the addition of other ingredients, such as polysaccharides.
Enzymatic peeling and discoloration of Red Bartlett pears
Authors: Ibarz Albert , Pagán Jordi , Garvín Alfonso , Ibarz Raquel , Gatius Ferran , Falguera Víctor 
Published: November 2012

SummaryIn purees manufacturing from some fruits, problems related to undesirable colours that come from fruit skin may arise. In this article, the feasibility of using an enzyme preparation to reduce the colour of Red Bartlett pears to be used for industrial purees has been studied. Kinetic models that suitably describe changing colour parameters with treatment time have been tested, seeking for the optimum ones. It was found that the most appropriated processing conditions for Red Bartlett pears discoloration are as follows: 45 °C with an enzyme concentration of 5000 ppm and a time that does not exceed 200 min. Although this treatment constitutes a solution for the problem of skin colour, a high concentration of enzyme and high treatment times are required, which makes the process expensive from the economic standpoint.
Polyphenol and volatile profiles of pomegranate (Punica granatum L.) fruit extracts and liquors
Authors: Galego Ludovina R. , Jockusch Steffen , Da Silva José P. 
Published: November 2012

SummaryThis work reports on the preparation of new aromatic pomegranate liquors by maceration of pomegranate juice and arils in Arbutus unedo L. distillate. The volatile, anthocyanin and ellagitannin profiles were monitored after each step. Aromatic compounds of pomegranates, such as limonene, 1‐hexenol and trans‐caryophyllene, were detected in trace levels and showed little contribution to the liquor aroma. The main volatiles of the liquors, namely isobutanol, isopentanols, vitispiranes and volatile esters, are given by the fruit distillate. The anthocyanin contents of the pomegranate juice were around 135 mg L−1 but decreased to about 5% of this value in the final liquors. Anthocyanin diglucoside compounds showed higher stability than their monoglucoside analogs. Punicalagins and punicalins showed contents between 80 and 135 mg L−1, depending on the used starting pomegranate material. These ellagitannins account for 60% of the total polyphenols. The procedure can be used to prepare aromatic pomegranate liquors with high polyphenol content.
Effects of polyphenol oxidation on in vitro iron availability in faba bean (Vicia faba L.) flour
Authors: Luo Yu‐Wei , Xie Wei‐Hua 
Published: November 2012

SummaryTannic acid (TA) and green tea extract (GTE) were added to faba bean flour, before and after incubation with polyphenol oxidase (mushroom tyrosinase), and the effect on in vitro iron availability was investigated. The inhibitory effect of TA and GTE was dose dependent, and the in vitro iron availability decreased from 64.5% in the pure faba bean flour to 9.8% (P < 0.05) with the addition of 1 mg of TA 1 g−1 to faba bean flour. The addition of 5 mg of GTE resulted in low in vitro iron availability as after the addition of 10 mg of TA (11.6% against 10.3% for 1 mg TA). Incubation of the polyphenols with tyrosinase before addition to the faba bean flour significantly increased the in vitro iron availability. The increase in iron availability was significant (P < 0.05) when amounts of 0.2, 0.5 or 1 mg g−1 TA or GTE added to faba bean flour, respectively, were oxidised, even with the lowest amount of tyrosinase (150 u). Oxidation of 1 mg TA or GTE with 150 u tyrosinase increased the in vitro iron availability from 10.3% to 15.5% and from 19.2% to 26.1%, respectively. At the 300‐u level, the addition of higher amounts of enzyme (from 600 to 900 u) did not have any effect. The results from the study therefore suggest that the oxidation of polyphenols may be a promising way to increase the availability of iron in polyphenol‐containing legume foods.
Optimisation of preservatives for dealcoholised red wine using a survival model for spoilage yeasts
Authors: Taboada‐Rodríguez Amaury , Belisario‐Sánchez Yulissa Y. , Cava‐Roda Rita , Cano Juan A. , López‐Gómez Antonio , Marín‐Iniesta Fulgencio 
Published: November 2012

SummaryTo optimise the application of preservatives in a new commercial dealcoholised red wine (DRW), a survival model of Rhodotorula mucilaginosa and Saccharomyces cerevisiae was developed. A factorial design included three variables, each at three levels: potassium sorbate, sodium metabisulphite and dimethyl dicarbonate (0, 100 and 200 mg L−1). DRW samples were inoculated with a cocktail of yeasts and counts were performed after 1, 2, 3, 4, 6 and 8 weeks of incubation. Logistic regression analysis was used to create survival models. Potassium sorbate and sodium metabisulphite did not show any fungicide effect alone, while the model predicts that dimethyl dicarbonate at 185 mg L−1 has fungicide activity. The interaction of potassium sorbate and dimethyl dicarbonate was statistically significant, so both could act as synergic preservatives. The logistic model obtained was validated with a 100% of accuracy for the conditions tested. Combining use of the studied preservatives reduced the individual dose for preservation of DRW.
Glucocorticoid receptor BclI polymorphism associates with immunomodulatory response to stress hormone in human peripheral blood mononuclear cells
Authors: Xiang L. , Marshall G. D. 
Published: November 2012

SummaryGlucocorticoid receptor (GR) variants have been found to be associated with stress‐related disorders. Our previous in vivo study revealed that the CC allele of GR BclI single‐nucleotide polymorphism (SNP) was more common in the high‐stress group, which had lower levels of both regulatory T cells (Treg) and Th1 cytokine. The current study was to investigate the associations between GR BclI polymorphism and immunomodulatory response to stress hormone in vitro in human peripheral blood mononuclear cells (PBMC). Blood samples were collected from 18 normal volunteers including 9 subjects with BclI polymorphism GG allele and 9 with wild‐type (WT) CC allele. PBMC were cultured with 10−8 m dexamethasone (DEX), which mimics the plasma cortisol level observed during periods of psychological stress for 24 h and 11 days. Gene expressions of transcription factors, stress hormone and cytokine receptors were analysed by real‐time RT‐PCR. FoxP3 mRNA was significantly altered in the BclI WT (decreased at 24 h and increased at 11 days) but not in the GG allele. GR mRNA was up‐regulated at 24 h and down‐regulated at 11 days in CC alleles (P < 0.01 and P < 0.05), rather than in GG alleles. The expression of β‐2 adrenergic receptor (β2AR) was increased at 24 h in both CC and GG alleles (P < 0.01 and P < 0.001), but decreased significantly at 11 days in only GG alleles. Expression of T‐bet and GATA‐3 was altered simultaneously in 24‐h culture with DEX from both groups. The BclI polymorphism of GR identifies different immunomodulatory responses to corticosteroids, which may explain, at least in part, the variability in individual sensitivity to stressors.
Focus on systemic lupus erythematosus in Indigenous Australians: towards a better understanding of autoimmune diseases
Authors: Vincent F. B. , Bourke P. , Morand E. F. , Mackay F. , Bossingham D. 
Published: November 2012

AbstractThe incidence and prevalence of autoimmune diseases such as rheumatoid arthritis, primary Sjögren syndrome, scleroderma and systemic lupus erythematosus (SLE) varies with geography and ethnicity. For example, SLE is reported to be more common in populations such as African‐Caribbeans and Indigenous Australians (IA). As well as socio‐economic status, variation in severity of disease may also show ethnic variability. The initial presentation of SLE in IA, in the context of a unique genetic background and distinctive environmental influences, is often florid with a recurring spectrum of clinical phenotypes. These clinical observations suggest a unique pathway for autoimmunity pathogenesis in this population. For instance, the high prevalence of bacterial infections in IA, particularly group A streptococcus, may be a potential explanation not only for increased incidence and prevalence of SLE but also the commonly florid acute disease presentation and propensity for rapidly progressive end organ threatening disease. This article will review the state of research in autoimmune disease of IA, consider key findings related to autoimmune disease in this population and propose a model potentially to explain the involvement of innate immunity and chronic infection in autoimmune disease pathogenesis. Ultimately, understanding of SLE at this level could affect management and result in personalised and targeted therapies to improve the health status of IA as well as better understanding of SLE pathogenesis per se.