bluuebunny/crossref_metadata_2025_split huggingface dataset:
Refolding, purification and crystallization of the FrpB outer membrane iron transporter fromNeisseria meningitidis
Authors: Saleem Muhammad , Prince Stephen M. , Patel Hema , Chan Hannah , Feavers Ian M. , Derrick Jeremy P. 
Published: January 2012

FrpB is an integral outer membrane protein from the human pathogenNeisseria meningitidis. It is a member of the TonB-dependent transporter family and promotes the uptake of iron across the outer membrane. There is also evidence that FrpB is an antigen and hence a potential component of a vaccine against meningococcal meningitis. FrpB incorporating a polyhistidine tag was overexpressed inEscherichia coliinto inclusion bodies. The protein was then solubilized in urea, refolded and purified to homogeneity. Two separate antigenic variants of FrpB were crystallized by sitting-drop vapour diffusion. Crystals of the F5-1 variant diffracted to 2.4 Å resolution and belonged to space groupC2, with unit-cell parametersa= 176.5,b= 79.4,c= 75.9 Å, β = 98.3°. Crystal-packing calculations suggested the presence of a monomer in the asymmetric unit. Crystals of the F3-3 variant also diffracted to 2.4 Å resolution and belonged to space groupP212121, with unit-cell parametersa= 85.3,b = 104.6,c= 269.1 Å. Preliminary analysis suggested the presence of an FrpB trimer in the asymmetric unit.
Crystallization and preliminary X-ray analysis of theClostridium botulinumtype D nontoxic nonhaemagglutinin
Authors: Miyata Keita , Inui Ken , Miyashita Shin-Ichiro , Sagane Yoshimasa , Hasegawa Kimiko , Matsumoto Takashi , Yamano Akihito , Niwa Koichi , Watanabe Toshihiro , Ohyama Tohru 
Published: January 2012

Clostridium botulinumproduces botulinum neurotoxin (BoNT) as a large toxin complex assembled with nontoxic nonhaemagglutinin (NTNHA) and/or haemagglutinin components. Complex formation with NTNHA is considered to be critical in eliciting food poisoning because the complex shields the BoNT from the harsh conditions in the digestive tract. In the present study, NTNHA was expressed inEscherichia coliand crystallized. Diffraction data were collected to 3.9 Å resolution. The crystal belonged to the trigonal space groupP321 orP3121/P3221, with unit-cell parametersa=b= 147.85,c= 229.74 Å. The structure of NTNHA will provide insight into the assembly mechanism that produces the unique BoNT–NTNHA complex.
The dynamic cytoskeleton of the developing male germ cell
Authors: Sperry Ann O. 
Published: January 2012

AbstractMammalian spermatogenesis is characterised by dramatic cellular change to transform the non‐polar spermatogonium into a highly polarised and functional spermatozoon. The acquisition of cell polarity is a requisite step for formation of viable sperm. The polarity of the spermatozoon is clearly demonstrated by the acrosome at the apical pole of the cell and the flagellum at the opposite end. Spermatogenesis consists of three basic phases: mitosis, meiosis and spermiogenesis. The final phase represents the period of greatest cellular change where cell‐type specific organelles such as the acrosome and the flagellum form, the nucleus migrates to the plasma membrane and elongates, chromatin condenses and residual cytoplasm is removed. An important feature of spermatogenesis is the change in the cytoskeleton that occurs throughout this pathway. In this review, the author will provide an overview of these transformations and provide insight into possible modes of regulation of these rearrangements during spermatogenesis. Although primary focus will be given to the microtubule cytoskeleton, the importance of actin filaments to the cellular transformation of the male germ cell will also be discussed.
Replication and fine‐mapping of a QTL for recurrent airway obstruction in European Warmblood horses
Authors: Shakhsi‐Niaei M. , Klukowska‐Rötzler J. , Drögemüller C. , Swinburne J. , Ehrmann C. , Saftic D. , Ramseyer A. , Gerber V. , Dolf G. , Leeb T. 
Published: January 2012

SummaryRecurrent airway obstruction (RAO), or ‘heaves’, is a common performance‐limiting allergic respiratory disease of mature horses. It is related to sensitization and exposure to mouldy hay and has a familial basis with a complex mode of inheritance. In a previous study, we detected a QTL for RAO on ECA 13 in a half‐sib family of European Warmblood horses. In this study, we genotyped additional markers in the family and narrowed the QTL down to about 1.5 Mb (23.7–25.2 Mb). We detected the strongest association with SNP BIEC2‐224511 (24 309 405 bp). We also obtained SNP genotypes in an independent cohort of 646 unrelated Warmblood horses. There was no genome‐wide significant association with RAO in these unrelated horses. However, we performed a genotypic association study of the SNPs on ECA 13 in these unrelated horses, and the SNP BIEC2‐224511 also showed the strongest association with RAO in the unrelated horses (praw = 0.00037). The T allele at this SNP was associated with RAO both in the family and the unrelated horses. Thus, the association study in the unrelated animals provides independent support for the previously detected QTL. The association study allows further narrowing of the QTL interval to about 0.5 Mb (24.0–24.5 Mb). We sequenced the coding regions of the genes in the critical region but did not find any associated coding variants. Therefore, the causative variant underlying this QTL is likely to be a regulatory mutation.
Understanding the evidence for and against the role of breastfeeding in allergy prevention
Authors: Matheson M. C. , Allen K. J. , Tang M. L. K. 
Published: January 2012

SummaryThe relationship between breastfeeding and allergic disease risk has been controversial. This article reviews the current evidence for the role of breastfeeding in the prevention of allergic disease. We found considerable methodological limitations inherent in most studies evaluating the effect of breastfeeding in allergic disease. Nevertheless, since randomized control trials in breast feeding research would be considered unethical, the evidence remains limited to poorer quality observational studies where participation and recall bias can severely affect the objectivity of the data collected. Furthermore, reporting of type of breastfeeding (exclusive, full or partial) may be biased by a participant's inherent belief system of what they think they should be doing. Current evidence is inconclusive regarding the effect of breastfeeding on the development of eczema, with the most recent systemic review reporting no protective effect. There is insufficient data regarding the effects of breastfeeding on objective measures of food allergy at any age. Studies show a paradoxical effect of breastfeeding on the prevention of asthma, with an apparent protective effect against early wheezing illness in the first years of life yet an increased risk of asthma in later life; however, these findings must be interpreted with caution. Existing studies fail to adequately adjust for confounders, including the critical issues of protection against early life respiratory illnesses and reverse causation. Therefore, it is possible that the effect of breastfeeding on early wheezing illness reflects protection against respiratory infection, the predominant trigger of wheezing in early childhood, rather than a true reduction in risk of asthma. In summary, future research that takes into account the potential contribution of confounding factors and effect modifiers is needed to clarify the role of breastfeeding in development of allergic disease and to inform current clinical guidelines on the prevention of allergic disease.
Immunohistochemical features of post‐radiation vaginal recurrences of endometrioid carcinomas of the endometrium: role for proteins involved in resistance to apoptosis and hypoxia
Authors: Santacana Maria , Yeramian Andree , Velasco Ana , Bergada Laura , Gatius Sonia , García Virginia , Azueta Ainara , Palacios Jose , Dolcet Xavier , Oliva Esther , Matias‐Guiu Xavier 
Published: January 2012

Santacana M, Yeramian A, Velasco A, Bergada L, Gatius S, García V, Azueta A, Palacios J, Dolcet X, Oliva E & Matias‐Guiu X  (2012) Histopathology 60, 460–471  Immunohistochemical features of post‐radiation vaginal recurrences of endometrioid carcinomas of the endometrium: role for proteins involved in resistance to apoptosis and hypoxiaAims:  Endometrioid carcinoma of the endometrium (EEC) is treated with surgery and radiotherapy. Post‐radiation recurrences are associated with increased risk of metastases. Comparison of the expression of genes important in the development and progression of EEC, and others involved in resistance to apoptosis and hypoxia and adaptation to radiation, was performed between post‐radiation vaginal recurrences (PVRs) and primary EECs. We tried to reproduce the results by exposing an EEC cell line to hypoxia and radiation.Methods and results:  Immunohistochemistry and tissue microarrays were used to compare 24 PVRs with 82 primary EECs. PVRs exhibited increased expression of p53 (P < 0.0001), cytoplasmic FLICE‐inhibitory protein (FLIP) (P < 0.0001), and Ki67 (P < 0.0001), and nuclear staining for FLIP, nuclear factor kappaB (NF‐κB) family members (p50, P < 0.0001; c‐Rel, P = 0.0077; RelB, P = 0.0157), and β‐catenin (P = 0.0001). Differences regarding p50, hypoxia‐inducible factor 1α (HIF‐1α), and cytoplasmic FLIP were statistically significant when PVRs and primary EECs were matched for histological grade. Exposure of the EEC cell line to hypoxia induced nuclear expression of β‐catenin, FLIP, and HIF‐1α, as well as increased NF‐κB activity. No changes in FLIP, HIF‐1α or NF‐κB were seen when cells were exposed to radiation. Nuclear expression of β‐catenin was seen at 3 Gy, but not at 1 Gy.Conclusions:  Genes involved in resistance to hypoxia are expressed in PVRs, and may play a role in the development of post‐radiation recurrences.
Patients with thymomas have an increased risk of developing additional malignancies: lack of immunological surveillance?
Authors: Granato Felice , Ambrosio Maria R , Spina Donatella , Lazzi Stefano , Rocca Bruno J , Voltolini Luca , Bongiolatti Stefano , Luzzi Luca , Gotti Giuseppe , Leoncini Lorenzo , Tosi Piero 
Published: January 2012

Granato F, Ambrosio M R, Spina D, Lazzi S, Rocca B J, Voltolini L, Bongiolatti S, Luzzi L, Gotti G, Leoncini L & Tosi P  (2012) Histopathology 60, 270–277  Patients with thymomas have an increased risk of developing additional malignancies: lack of immunological surveillance?Aims:  To assess the risk, for patients with thymoma, of developing an additional malignancy (AM).Methods and results:  We studied 68 patients with thymomas. Based on the World Health Organization classification, the tumours were categorised as A, AB or B (B1, B2, B3) thymomas. Control populations comprised 114 patients with colorectal cancer, 108 patients with lymphoma and 123 patients with thyroid carcinoma. Patients with thymomas showed a higher risk of developing an AM (22 of 68 patients versus 11 of 114, eight of 108, and eight of 123 patients, respectively; P = 0.0002). The association between thymomas and AMs was related to the thymoma histotype, with B1, B2, B3 and AB tumours showing a higher risk of developing an AM than A thymomas (P = 0.0474).Conclusions:  Patients affected by thymomas showed a significantly higher risk of developing additional malignancies than those in the control groups, and cases that exhibited a predominantly cortical component were more likely to develop other neoplasms. This may be related to the functions of cortical thymic epithelial cells in providing for T lymphocyte maturation through interaction with major histocompatibility complexes.
Immunohistochemical analysis of hepatocyte growth factor and c‐Met in plasma cell disease
Authors: Wader Karin F , Fagerli Unn‐Merete , Børset Magne , Lydersen Stian , Hov Håkon , Sundan Anders , Bofin Anna , Waage Anders 
Published: January 2012

Wader K F, Fagerli U‐M, Børset M, Lydersen S, Hov H, Sundan A, Bofin A & Waage A  (2012) Histopathology60,443–451  Immunohistochemical analysis of hepatocyte growth factor and c‐Met in plasma cell diseaseAims:  Interaction with the bone marrow microenvironment is important for homing and survival of myeloma cells. One cytokine involved in this process is hepatocyte growth factor (HGF). HGF, by binding to the receptor tyrosine kinase c‐Met, mediates a broad range of tumour progression activities. Our aims were to investigate whether HGF and c‐Met are present in bone marrow and extramedullary tumours from patients with monoclonal plasma cell disease, and whether c‐Met is activated.Methods and results:  Expression of HGF, c‐Met and phospho‐c‐Met was studied by immunohistochemistry in biopsies from 80 patients with monoclonal plasma cell disease. Cytoplasmic staining for HGF in plasma cells was demonstrated in 58 of 68 biopsies from multiple myeloma patients (85%), but also in biopsies from nine of 10 healthy individuals. Membranous staining for c‐Met was found in 25 of 63 multiple myeloma patients (40%) and in none of 10 healthy individuals. Membranous staining for phospho‐c‐Met was found in biopsies from 15 of 21 c‐Met‐positive myeloma patients (71%).Conclusions:  Our data point to c‐Met expression as one of the factors that distinguishes normal from malignant plasma cells, and indicate that the HGF/c‐Met system is activated in multiple myeloma patients.
HMGA1 and HMGA2 protein expression correlates with advanced tumour grade and lymph node metastasis in pancreatic adenocarcinoma
Authors: Piscuoglio Salvatore , Zlobec Inti , Pallante Pierlorenzo , Sepe Romina , Esposito Francesco , Zimmermann Arthur , Diamantis Ioannis , Terracciano Luigi , Fusco Alfredo , Karamitopoulou Eva 
Published: January 2012

Piscuoglio S, Zlobec I, Pallante P, Sepe R, Esposito F, Zimmermann A, Diamantis I, Terracciano L, Fusco A & Karamitopoulou E  (2012) Histopathology 60, 397–404  HMGA1 and HMGA2 protein expression correlates with advanced tumour grade and lymph node metastasis in pancreatic adenocarcinomaAims:  Pancreatic ductal adenocarcinoma follows a multistep model of progression through precursor lesions called pancreatic intraepithelial neoplasia (PanIN). The high mobility group A1 (HMGA1) and high mobility group A2 (HMGA2) proteins are architectural transcription factors that have been implicated in the pathogenesis and progression of malignant tumours, including pancreatic cancer. The aim of this study was to explore the role of HMGA1 and HMGA2 in pancreatic carcinogenesis.Methods and results:  HMGA1 and HMGA2 expression was examined in 210 ductal pancreatic adenocarcinomas from resection specimens, combined on a tissue microarray also including 40 examples of PanIN and 40 normal controls. The results were correlated with the clinicopathological parameters of the tumours and the outcome of the patients. The percentage of tumour cells showing HMGA1 and HMGA2 nuclear immunoreactivity correlated positively with increasing malignancy grade and lymph node metastasis. Moreover, HMGA1 and HMGA2 expression was significantly higher in invasive carcinomas than in PanINs. No, or very low, expression was found in normal pancreatic tissue.Conclusions:  Our results suggest that HMGA1 and HMGA2 are implicated in pancreatic carcinogenesis and may play a role in tumour progression towards a more malignant phenotype.
Gastro‐oesophageal reflux disease is associated with up‐regulation of desmosomal components in oesophageal mucosa
Authors: Wex Thomas , Mönkemüller Klaus , Stahr Antje , Kuester Doerthe , Fry Lucia C , Völkel Simone , Kandulski Arne , Roessner Albert , Malfertheiner Peter 
Published: January 2012

Wex T, Mönkemüller K, Stahr A, Kuester D, Fry L C, Völkel S, Kandulski A, Roessner A & Malfertheiner P  (2012) Histopathology 60, 405–415  Gastro‐oesophageal reflux disease is associated with up‐regulation of desmosomal components in oesophageal mucosaAims:  Gastro‐oesophageal reflux disease (GERD) is associated with impaired epithelial barrier function. This study was aimed at investigating the role of desmosomal proteins in relation to GERD.Methods and results:  Ninety‐five patients with GERD‐related symptoms (erosive, n = 51; non‐erosive, n = 44) and 27 patients lacking those symptoms were included. Endoscopic and histological characterization of oesophagitis was performed according to the Los Angeles and Ismeil–Beigi criteria, respectively. Multiple biopsies were taken from the oesophageal mucosa of each patient. Gene expression analysis of plakoglobin, desmoglein‐1, desmoglein‐2 and desmoglein‐3 was performed by quantitative real time (RT)‐polymerase chain reaction and immunohistochemistry in the oesophageal mucosa. Routine histology revealed specific GERD‐related alterations, such as dilatation of intercellular spaces (DIS), basal cell hyperplasia (BCH), and elongation of the papillae, in the oesophageal mucosa of patients with GERD, as compared with controls (all parameters: P < 0.05). All four genes and corresponding proteins were found to be up‐regulated by between 1.7 and 8.1‐fold (transcript level, P < 0.05; protein level, P < 0.05). Induced gene expression levels of plakoglobin, desmoglein‐1 and desmoglein‐2 correlated significantly with DIS and BCH.Conclusions:  Taken together, the uniform up‐regulation of desmosomal genes/proteins in the oesophageal mucosa of patients with GERD supports the concept of architectural and molecular changes in the desmosomal compartment in the pathogenesis of GERD.
A simple immunohistochemical algorithm predicts the risk of distant metastases in right‐sided colon cancer
Authors: Neumann Jens , Horst David , Kriegl Lydia , Maatz Susanne , Engel Jutta , Jung Andreas , Kirchner Thomas 
Published: January 2012

Neumann J, Horst D, Kriegl L, Maatz S, Engel J, Jung A & Kirchner T  (2012) Histopathology 60, 416–426  A simple immunohistochemical algorithm predicts the risk of distant metastases in right‐sided colon cancerAims:  A test predicting distant metastases would be valuable for prognostication in colon cancer (CC). In previous studies, CC with microsatellite instability (MSI) showed a reduced risk of distant metastases. High expression of CD133 and β‐catenin, both related to cancer stem cell phenotypes, might be predictive markers for metastasis. The aim of this study was to develop a simple and robust test for risk assessment of distant metastases in CC.Methods and results:  In a case–control study, 57 cases of right‐sided CC specimens with synchronous distant metastases were matched with 57 CC without distant metastases. Immunohistochemistry for MLH1, CD133 and nuclear β‐catenin was carried out. To define the diagnostic algorithm the tumours were first stratified according to their MLH1 expression. Loss of MLH1 expression was correlated significantly with a very low risk of distant metastases (5.3%; P = 0.00003). In MLH1‐positive cases, combined high scores of CD133 and β‐catenin were associated with a very high rate of distant metastases (94.4%), whereas the risk was intermediate for carcinomas with either low CD133 and/or low β‐catenin expression (P = 0.0007). A validation study using an independent set of 68 right‐sided CC specimens showed a clear trend towards risk stratification according to the algorithm; however, sample sizes were small, and associations were not statistically significant.Conclusions:  By the use of three markers, this algorithm allowed identification of subgroups of right‐sided CC patients with extremely high and extremely low risk of distant metastases.
Performance of fluorescence‐based and conventional methods of occlusal caries detection in primary molars – an in vitro study
Authors: NOVAES TATIANE F. , MATOS RONILZA , GIMENEZ THAIS , BRAGA MARIANA M. , DE BENEDETTO MONIQUE S. , MENDES FAUSTO M. 
Published: January 2012

International Journal of Paediatric Dentistry 2012; 22: 459–466Aim.  This in vitro study aimed to test the performance of fluorescence‐based methods in detecting occlusal caries lesions in primary molars compared to conventional methods.Design.  Two examiners assessed 113 sites on 77 occlusal surfaces of primary molars using three fluorescence devices: DIAGNOdent (LF), DIAGNOdent pen (LFpen), and fluorescence camera (VistaProof‐FC). Visual inspection (ICDAS) and radiographic methods were also evaluated. One examiner repeated the evaluations after one month. As reference standard method, the lesion depth was determined after sectioning and evaluation in stereomicroscope. The area under the ROC curve (Az), sensitivity, specificity, and accuracy of the methods were calculated at enamel (D1) and dentine caries (D3) lesions thresholds. The intra and interexaminer reproducibility were calculated using the intraclass correlation coefficient (ICC) and kappa statistics.Results.  At D1, visual inspection presented higher sensitivities (0.97–0.99) but lower specificities (0.18–0.25). At D3, all the methods demonstrated similar performance (Az values around 0.90). Visual and radiographic methods showed a slightly higher specificity (values higher than 0.96) than the fluorescence based ones (values around 0.88). In general, all methods presented high reproducibility (ICC higher than 0.79).Conclusions.  Although fluorescence‐based and conventional methods present similar performance in detecting occlusal caries lesions in primary teeth, visual inspection alone seems to be sufficient to be used in clinical practice.
Microleakage and penetration of a hydrophilic sealant and a conventional resin‐based sealant as a function of preparation techniques: a laboratory study
Authors: KHOGLI AHMED ELTIGANI , CAUWELS RITA , VERCRUYSSE CHRIS , VERBEECK RONALD , MARTENS LUC 
Published: January 2012

International Journal of Paediatric Dentistry 2013; 23: 13–22
Background.  Optimal pit and fissure sealing is determined by surface preparation techniques and choice of materials.Aim.  This study aimed (i) to compare the microleakage and penetration depth of a hydrophilic sealant and a conventional resin‐based sealant using one of the following preparation techniques: acid etching (AE) only, a diamond bur + AE, and Er:YAG laser combined with AE, and (ii) to evaluate the microleakage and penetration depth of the hydrophilic pit and fissure sealant on different surface conditions.Design.  Eighty recently extracted 3rd molars were randomly assigned to eight groups of ten teeth according to the material, preparation technique, and surface condition. For saliva contamination, 0.1 mL of fresh whole human saliva was used. All samples were submitted to 1000 thermal cycles and immersed in 2% methylene blue dye for 4 h. Sections were examined by a light microscope and analysed using image analysis software (Sigmascan®).Results.  The combination of Er:YAG + AE + conventional sealant showed the least microleakage. The sealing ability of the hydrophilic sealant was influenced by the surface condition.Conclusion.  Er:YAG ablation significantly decreased the microleakage at the tooth–sealant interface compared to the non‐invasive technique. The hydrophilic sealant applied on different surface conditions showed comparable result to the conventional resin‐based sealant.