bluuebunny/crossref_metadata_2025_split huggingface dataset:
Single‐step affinity purification of recombinant proteins using a self‐excising module from Neisseria meningitidis FrpC
Authors: Sadilkova Lenka , Osicka Radim , Sulc Miroslav , Linhartova Irena , Novak Petr , Sebo Peter 
Published: July 2008

AbstractPurification of recombinant proteins is often a challenging process involving several chromatographic steps that must be optimized for each target protein. Here, we developed a self‐excising module allowing single‐step affinity chromatography purification of untagged recombinant proteins. It consists of a 250‐residue‐long self‐processing module of the Neisseria meningitidis FrpC protein with a C‐terminal affinity tag. The N terminus of the module is fused to the C terminus of a target protein of interest. Upon binding of the fusion protein to an affinity matrix from cell lysate and washing out contaminating proteins, site‐specific cleavage of the Asp–Pro bond linking the target protein to the self‐excising module is induced by calcium ions. This results in the release of the target protein with only a single aspartic acid residue added at the C terminus, while the self‐excising affinity module remains trapped on the affinity matrix. The system was successfully tested with several target proteins, including glutathione‐S‐transferase, maltose‐binding protein, β‐galactosidase, chloramphenicol acetyltransferase, and adenylate cyclase, and two different affinity tags, chitin‐binding domain or poly‐His. Moreover, it was demonstrated that it can be applied as an alternative to two currently existing systems, based on the self‐splicing intein of Saccharomyces cerevisiae and sortase A of Staphylococcus aureus.
Modulation of kinase‐inhibitor interactions by auxiliary protein binding: Crystallography studies on Aurora A interactions with VX‐680 and with TPX2
Authors: Zhao Baoguang , Smallwood Angela , Yang Jingsong , Koretke Kristin , Nurse Kelvin , Calamari Amy , Kirkpatrick Robert B. , Lai Zhihong 
Published: July 2008

AbstractVX‐680, also known as MK‐0457, is an ATP‐competitive small molecule inhibitor of the Aurora kinases that has entered phase II clinical trials for the treatment of cancer. We have solved the cocrystal structure of AurA/TPX2/VX‐680 at 2.3 Å resolution. In the crystal structure, VX‐680 binds to the active conformation of AurA. The glycine‐rich loop in AurA adopts a unique bent conformation, forming a π–π interaction with the phenyl group of VX‐680. In contrast, in the published AurA/VX‐680 structure, VX‐680 binds to AurA in the inactive conformation, interacting with a hydrophobic pocket only present in the inactive conformation. These data suggest that TPX2, a protein cofactor, can alter the binding mode of VX‐680 with AurA. More generally, the presence of physiologically relevant cofactor proteins can alter the kinetics, binding interactions, and inhibition of enzymes, and studies with these multiprotein complexes may be beneficial to the discovery and optimization of enzyme inhibitors as therapeutic agents.
Structure of AscE and induced burial regions in AscE and AscG upon formation of the chaperone needle‐subunit complex of type III secretion system in Aeromonas hydrophila
Authors: Tan Yih Wan , Yu Hong Bing , Leung Ka Yin , Sivaraman J. , Mok Yu‐Keung 
Published: July 2008

AbstractIn the type III secretion system (T3SS) of Aeromonas hydrophila, the putative needle complex subunit AscF requires both putative chaperones AscE and AscG for formation of a ternary complex to avoid premature assembly. Here we report the crystal structure of AscE at 2.7 Å resolution and the mapping of buried regions of AscE, AscG, and AscF in the AscEG and AscEFG complexes using limited protease digestion. The dimeric AscE is comprised of two helix–turn–helix monomers packed in an antiparallel fashion. The N‐terminal 13 residues of AscE are buried only upon binding with AscG, but this region is found to be nonessential for the interaction. AscE functions as a monomer and can be coexpressed with AscG or with both AscG and AscF to form soluble complexes. The AscE binding region of AscG in the AscEG complex is identified to be within the N‐terminal 61 residues of AscG. The exposed C‐terminal substrate‐binding region of AscG in the AscEG complex is induced to be buried only upon binding to AscF. However, the N‐terminal 52 residues of AscF remain exposed even in the ternary AscEFG complex. On the other hand, the 35‐residue C‐terminal region of AscF in the complex is resistant to protease digestion in the AscEFG complex. Site‐directed mutagenesis showed that two C‐terminal hydrophobic residues, Ile83 and Leu84, of AscF are essential for chaperone binding.
Recruitment variability of resident brown trout in peripheral populations from southern Europe
Authors: NICOLA GRACIELA G. , ALMODÓVAR ANA , JONSSON BROR , ELVIRA BENIGNO 
Published: July 2008

Summary1. Population regulation was studied for seven consecutive years (1992–98) in five rivers at the periphery of the distribution ofSalmo trutta, where the fish were living under environmental constraints quite different from those of the main distribution area.2. Recruitment is naturally highly variable and the populations had been earlier classified as overexploited. Thus we expected that densities of young trout in most populations would be too low for density‐dependent mortality to operate. We tested this by fitting the abundance of recruits to egg densities over seven consecutive years (stock–recruitment relationship), and used the results to judge whether exploitation should be restricted in the interests of conserving the populations.3. The density of 0+ trout in early September, as well as the initial density of eggs and parents, varied greatly among localities and years. The data for all populations fitted the Ricker stock–recruitment model. The proportion of variance explained by the population curves varied between 32% and 51%. However, in most cases the observations were in the density‐independent part of the stock–recruitment curve, where densities of the recruits increased proportionally with egg densities.4. Our findings suggest that recruitment densities in most rivers and years were below the carrying capacity of the habitats. Although density‐dependent mechanisms seemed to regulate fish abundance in some cases, environmental factors and harvesting appeared generally to preclude populations from reaching densities high enough for negative feedbacks to operate. The findings thus lend support toHaldane’s (1956)second hypothesis that changes in population density are primarily due to density‐independent factors in unfavourable areas and areas with low density due to exploitation. Exploitation should be reduced to allow natural selection to operate more effectively.
Transcription factors involved in pancreas development are expressed in paediatric solid pseudopapillary tumours
Authors: Galmiche L , Sarnacki S , Verkarre V , Boizet B , Duvillie B , Fabre M , Jaubert F 
Published: July 2008

Aims:  Solid pseudopapillary tumours (SPT) are rare pancreatic tumours, especially in children. The origin of this benign tumour remains unknown. Mutations of β‐catenin, a gene essential for pancreatic development, are constantly found, leading to delocalization of immunohistochemical signals from the cytoplasm to the nuclei of tumour cells. The aim was to report clinical and histological data of eight children with SPT and explore the immunohistochemical expression of pancreatic duodenal homeobox (PDX) 1 and Sox9, known to be crucial for pancreatic development and linked to the β‐catenin cascade.Methods and results:  Eight children with features suggestive of SPT underwent surgical resection. Tumours displayed typical histological appearances. One was incompletely resected and recurred. Immunolabelling revealed nuclear location of β‐catenin in all cases and strong cytoplasmic but no nuclear expression of PDX1 or Sox9 in all but one case.Conclusions:  The clinical behaviour of SPT in the paediatric population is similar to its adult counterpart. Complete surgical resection is essential. PDX1 and Sox9 proteins are exclusively expressed in the cytoplasmic compartment in SPT, suggesting overexpression of the corresponding genes linked to β‐catenin mutations. These findings favour the hypothesis that SPT originates from transformation of normally quiescent pancreatic stem cells.
The effect of par‐baking and frozen storage time on the quality of cup cake
Authors: Karaoğlu Mehmet Murat , Kotancilar Halis Gürbüz , Gerçekaslan Kamil Emre 
Published: July 2008

SummaryThe effects of frozen storage and initial baking time of par‐baked cake on baking loss, volume, moisture, colour and textural properties of cake obtained after thawing and rebaking were investigated. Cakes, par‐baked at 175 °C for 15, 20 and 25 min, were stored at −18 °C for 3, 6 and 9 months. After storage, par‐baked cakes were thawed and rebaked at 175 °C for 10, 15 and 20 min. Baking loss, moisture content, L and +b colour values, firmness, gumminess and chewiness of the resulting full‐baked cakes were significantly affected by both par‐baking and frozen storage time, while specific volume, cohesiveness, springiness and resilience values were significantly affected by frozen storage time. The increase in the time of frozen storage of the par‐baked cake leads to a decrease in the quality of the rebaked cake, namely an increase of baking loss and cake crumb firmness, and a loss in the moisture content and specific volume. Moisture of cake crumb, L and +b colour values, firmness, gumminess and chewiness significantly increased as the par‐baking time increased. However, regarding baking loss, specific volume, moisture content and textural properties, 3‐month intermediate storage at −18 °C and 20‐min initial baking time gave the best result among the cakes produced by using the two‐step baking procedure.
Comparison of rheological models for determining dark chocolate viscosity
Authors: Afoakwa Emmanuel Ohene , Paterson Alistair , Fowler Mark , Vieira Joselio 
Published: July 2008

SummaryParameters in chocolate rheology, namely shear viscosity and yield stress, are important in manufacture and directly influenced by product particle size distribution (PSD) and composition. The Casson model was the standard confectionery industry strategy to quantify rheological properties of molten chocolate until in 2000, the International Confectionery Association recommended the use of interpolation data to describe viscosity. The two strategies are compared and correlated in defining rheological properties of molten dark chocolates prepared using different PSD, fat and lecithin content. Rheological parameters were determined using a shear rate‐controlled rheometer and data examined using correlation, regression and principal component analyses to establish their inter‐relationships. Correlation and regression analyses showed high correlation (r = 0.89–1.00) and regression coefficients (R2 = 0.84–1.00). The newer International Confectionery Association technique gave higher correlation and regression coefficients than the Casson model, but multivariate principal component analysis showed that the two models were highly related and either could effectively quantify dark chocolate viscosity parameters.
Ethical dilemmas of a large national multi‐centre study in Australia: time for some consistency
Authors: Driscoll Andrea , Currey Judy , Worrall‐Carter Linda , Stewart Simon 
Published: July 2008

Aims and objectives.  To examine the impact and obstacles that individual Institutional Research Ethics Committee (IRECs) had on a large‐scale national multi‐centre clinical audit called the National Benchmarks and Evidence‐based National Clinical guidelines for Heart failure management programmes Study.Background.  Multi‐centre research is commonplace in the health care system. However, IRECs continue to fail to differentiate between research and quality audit projects.Methods.  The National Benchmarks and Evidence‐based National Clinical guidelines for Heart failure management programmes study used an investigator‐developed questionnaire concerning a clinical audit for heart failure programmes throughout Australia. Ethical guidelines developed by the National governing body of health and medical research in Australia classified the National Benchmarks and Evidence‐based National Clinical guidelines for Heart failure management programmes Study as a low risk clinical audit not requiring ethical approval by IREC.Results.  Fifteen of 27 IRECs stipulated that the research proposal undergo full ethical review. None of the IRECs acknowledged: national quality assurance guidelines and recommendations nor ethics approval from other IRECs. Twelve of the 15 IRECs used different ethics application forms. Variability in the type of amendments was prolific. Lack of uniformity in ethical review processes resulted in a six‐ to eight‐month delay in commencing the national study.Conclusions.  Development of a national ethics application form with full ethical review by the first IREC and compulsory expedited review by subsequent IRECs would resolve issues raised in this paper. IRECs must change their ethics approval processes to one that enhances facilitation of multi‐centre research which is now normative process for health services.Relevance to clinical practice.  The findings of this study highlight inconsistent ethical requirements between different IRECs. Also highlighted are the obstacles and delays that IRECs create when undertaking multi‐centre clinical audits. However, in our clinical practice it is vital that clinical audits are undertaken for evaluation purposes. The findings of this study raise awareness of inconsistent ethical processes and highlight the need for expedient ethical review for clinical audits.
Nurse supplementary prescribing for patients with diabetes: a national questionnaire survey
Authors: Carey Nicola , Courtenay Molly 
Published: July 2008

Aims and objectives.  To examine the prescribing practices of nurse supplementary prescribing in diabetes.Background.  Nurses in several roles are involved in the management of medicines for patients with diabetes. Nurse prescribing should help optimise these roles. Nurses in the UK have virtually the same independent prescribing rights as doctors. There is little or no evidence on the extent to which nurse supplementary prescribing is used, or the impact and activity of nurse supplementary prescribing for patients with diabetes.Design.  Survey.Method.  A random sample of 214 nurse supplementary prescribers self‐completed a written questionnaire.Results.  The majority of nurses held an academic qualification at degree level or higher, had a wealth of clinical experience, worked full‐time, were based in primary care and worked in general practice. The majority of nurses prescribed between one and five items a week. Oral anti‐diabetic drugs, hypertension and lipid‐regulating drugs and insulins were the products most often prescribed. Over 85% had undertaken specialist training in diabetes prior to undertaking the prescribing programme.Conclusion.  Supplementary prescribing provides a practical and useful framework within which to prescribe medicines for patients with diabetes and its associated complications. Specialist diabetes training is a necessary prerequisite for nurses prescribing in this area. It is evident that there is still a place for supplementary prescribing.Implications for clinical practice.  • Recent legislative changes mean that nurses can now independently prescribe practically any drug.• Nurses in general practice appear to prescribe most frequently as a nurse supplementary prescriber for patients with diabetes.• Nurse supplementary prescribers are likely to use this mode of prescribing to deliver medicines to patients with diabetes.• Over two‐thirds prescribe for common but serious complications of diabetes, e.g. hypertension, hyperlipidaemia and cardiovascular disease
Regulation of glucocorticoid receptor in nasal polyps by systemic and intranasal glucocorticoids
Authors: Pujols L. , Alobid I. , Benítez P. , Martínez‐Antón A. , Roca‐Ferrer J. , Fokkens W. J. , Mullol J. , Picado C. 
Published: July 2008

Background:  Poor response of nasal polyps to glucocorticoids (GCs) may be because of abnormal expression of GC receptors (GR) α and β or to downregulation of GRα. We aimed to evaluate the in vivo regulation of GR isoforms in GC‐treated nasal polyps and to assess the relationship between clinical response to GCs and GR levels.Methods:  Patients with nasal polyps were randomly (3:1) treated (n = 51) or not (n = 14) with oral prednisone and intranasal budesonide for 2 weeks, plus intranasal budesonide for 10 additional weeks. Nasal symptoms were evaluated. Biopsies were obtained before (w0) and after 2 (w2) and 12 (w12) weeks of treatment, and analysed for their inflammatory content and GR mRNA (102 cDNA copies/μg total RNA) and protein (% immunoreactive inflammatory cells) expression. Healthy nasal mucosa (n = 11) was also investigated. Data are presented as median and 25–75th percentile.Results:  At w0, nasal polyps expressed less GRα mRNA (1343;683–2263; P < 0.05) and GR protein (41;29–54; P < 0.05) than nasal mucosa (2474;1346–2933; 60;51–72, respectively). GRβ immunoreactivity was higher in nasal polyps (11;4–19; P < 0.05) than in nasal mucosa (5;2–5). At w2, increased GRα mRNA (2010;1037–2732; P < 0.01) and GR protein (56;27–71; P = 0.056) were found compared with w0 (1177;759–2058; 37;29–55, respectively). At w12, GRα mRNA and GR protein were similar to w0. GRβ expression was unaltered by treatment. Neither GRα nor GRβ correlated with nasal symptoms. GR immunoreactivity negatively correlated with eosinophils (r = −0.478; P < 0.001).Conclusions:  GRα is downregulated in nasal polyps and upregulated by GC treatment. Neither GRα nor GRβ appear to determine the sensitivity to GCs in nasal polyposis.
Quantitative Trait Loci for Resistance to Powdery Mildew in Cucumber under Seedling Spray Inoculation and Leaf Disc Infection
Authors: Longzhou Liu , Xiaojun Yuan , Run Cai , Junsong Pan , Huanle He , Lihua Yuan , Yuan Guan , Lihuang Zhu 
Published: July 2008

AbstractPowdery mildew (PM) is an economically important disease of cucumber (Cucumis sativus L.) caused by the obligate biotrophic pathogen, Podosphaera xanthii, in greenhouse and field. In this study, a resistant breeding line S06 (greenhouse European type) and a susceptible cucumber inbred line S94 (Northern China open‐field type) and their F2:3 populations were used to investigate PM resistance under seedling spray inoculation in two growing seasons and leaf disc infection in one season. Quantitative trait loci (QTL) analysis was undertaken based on a constructed molecular linkage map of the corresponding F2 population using composite interval mapping. Five QTL (PM1.1, PM1.2, PM2.1, PM2.2 and PM5.1) for PM resistance were identified and located on linkage groups (LG) 1, 2 and 5, explaining 3.4–45.0% of the phenotypic variation. Three consistent QTL (PM1.1, PM2.1 and PM5.1) were detected under the three test conditions. The present results of the QTL analysis for PM resistance have laid a basis for breeding PM‐resistant cucumber varieties by molecular marker‐assisted selection.
Sporulation, Inoculation Methods and Pathogenicity of Ustilaginoidea albicans, the Cause of White Rice False Smut in China
Authors: Wang Shu , Li Min , Dong Hai , Liu Xiao‐Zhou , Bai Yuan‐Jun , Yang Hao 
Published: July 2008

AbstractSporulation and inoculation methods were studied to determine the pathogenicity of Ustilaginoidea albicans S Wang and JK Bai, the cause of white rice false smut, isolated in China. High sporulation was obtained using solid culture, liquid fermentation and double layer incubation system. In liquid culture, the culture media composition contributed to observed differences in sporulation. Potato sucrose broth was the optimal media to promote conidia production, increasing the concentration to up to 4.8 × 108 spores/ml. Inoculation by injection and spraying resulted in disease development, demonstrating that conidia of white rice false smut are virulent to rice plants, despite apparent differences in disease incidence. Estimates of mean disease severity were higher for injection than for spraying as assessed by percent infected panicles. The two methods resulted in disease severity ranging from 12.0 to 28.7% and 8.2 to 20.7%, respectively. Low temperature exposure after inoculation had a strong stimulatory effect on disease development.
Differences in multidisciplinary team assessment and treatment between patients with stage IV colon and rectal cancer
Authors: Segelman J. , Singnomklao T. , Hellborg H. , Martling A. 
Published: July 2008

AbstractObjective  At diagnosis, 14–27% of patients with colorectal cancer (CRC) have distant metastases (stage IV) and a poor prognosis. Today, treatment decisions for CRC patients are often made at multidisciplinary team (MDT) conferences. The aim of this study was to evaluate the effects of development and implementation of MDT assessment and treatment in patients with stage IV colon cancer (CC) and rectal cancer (RC) in a large population.Method  All 1449 patients who had stage IV CRC at the time of diagnosis and were registered in the regional quality registry of Stockholm from 1995 to 2004 were included. Patients with CC and RC were grouped according to treatment and their characteristics were analysed separately.Results  In total, 1000 patients with CC and 449 patients with RC had stage IV disease. Of these, 689 (68.9%) CC patients and 352 (78.4%) RC patients were assessed by a MDT and the proportion increased over the study period (P < 0.001). Surgery for metastases was undertaken on 39 (3.9%) CC patients and 38 (8.5%) RC patients (P < 0.001). CRC patients selected for metastasis surgery had 37% 5‐year survival when compared with 2% in patients who were not selected for metastasis surgery (P < 0.001).Conclusion  Patients with CC were less often assessed by a MDT and less often had metastasis surgery than RC patients. The proportion of patients with CC and RC assessed by a MDT increased during the study period, as did the proportion who had surgery for metastases. MDT assessment opens up the opportunity for more aggressive treatment with better outcomes.
Adenocarcinoma complicating restorative proctocolectomy for ulcerative colitis with mucosectomy performed by Cavitron Ultrasonic Surgical Aspirator®
Authors: Branco B. C. , Sachar D. B. , Heimann T. , Sarpel U. , Harpaz N. , Greenstein A. J. 
Published: July 2008

AbstractThis is a report of adenocarcinoma arising in an ileal pouch after restorative proctocolectomy (RPC) with rectal mucosal stripping performed by Cavitron Ultrasonic Surgical Aspirator (CUSA®) for ulcerative colitis. The CUSA® was introduced to simplify and optimize ileal pouch–anal anastomosis with mucosectomy and has been shown to shorten the operative time and reduce blood loss. Its use however, may increase the number of pathology specimens made uninterpretable on account of tissue ablation. In the present case, even though preoperative colonoscopy had clearly shown dysplasia, the surgical pathology report could not detect any neoplasia in the specimen; hence, the patient was not surveyed for pouch cancer. Six years later, the patient presented with intestinal obstruction caused by cancer. While protocols for universal pouch surveillance remain somewhat controversial, we conclude on the basis of this case and a review of the literature that in RPC with mucosectomy performed by CUSA®, pouch cancer surveillance is particularly important because remnants of rectal epithelium may have been left behind and tissue ablation may have made the surgical pathology report uninterpretable.
Blow as well as pull: an innovative technique for dealing with a rectal foreign body.
Authors: Manimaran N. , Shorafa M. , Eccersley J. 
Published: July 2008

AbstractRemoval of a rectal foreign body (FB) has become increasingly common in the recent times. Nature of rectal FB is limited only by the imagination of the patient concerned. Many techniques have been described for removal and various theories have been put forward to explain the mechanics of the procedure. We report a case where a new technique had been employed.