bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibody validation by combining immunohistochemistry and protein extraction from formalin‐fixed paraffin‐embedded tissues
Authors: Schuster Claudia , Malinowsky Katharina , Liebmann Sibylle , Berg Daniela , Wolff Claudia , Tran Kai , Schott Christina , Reu Simone , Neumann Jens , Faber Claudius , Höfler Heinz , Kirchner Thomas , Becker Karl‐Friedrich , Hlubek Falk 
Published: March 2012

Schuster C, Malinowsky K, Liebmann S, Berg D, Wolff C, Tran K, Schott C, Reu S, Neumann J, Faber C, Höfler H, Kirchner T, Becker K‐F & Hlubek F(2012) Histopathology 60, E37–E50Antibody validation by combining immunohistochemistry and protein extraction from formalin‐fixed paraffin‐embedded tissuesAims:  Personalized cancer treatment strategies depend on comprehensive and detailed characterization of individual human malignancies. Clinical pathology, particularly immunohistochemical evaluation of biomarkers in tissues, is considered to be the approved standard for diagnostic and therapeutic decisions, having a direct influence on patient management and therapy. Although antibody‐based approaches are established and integrated successfully into both clinical and research applications, for personalized treatment regimens new demands have been placed on the quality, reproducibility and accuracy of antibody‐based assays. To ensure the accuracy of specific antigen detection in immunohistochemistry, we introduce a novel approach for antibody validation.Methods and results:  In a tandem approach we used the same archival tissue of interest for antibody validation by combining extraction of immunoreactive proteins from formalin‐fixed, paraffin‐embedded tissue with Western blot analysis and immunohistochemistry. This procedure allows for specification of the antigen detected and for localization of the protein in the tissue. Of the 32 antibodies tested used in research and routine diagnostics, 19 showed reliable specificity in both assays.Conclusion:  This study emphasizes the advantage of combining suitable methods to ensure reproducibility and specific antigen detection. Based on our results, we propose a novel step‐by‐step strategy to validate antibody specificity and reduce variability of immunohistochemical results.
CD133 expression is associated with poor outcome in neuroblastoma via chemoresistance mediated by the AKT pathway
Authors: Sartelet Hervé , Imbriglio Tina , Nyalendo Carine , Haddad Elie , Annabi Borhane , Duval Michel , Fetni Raouf , Victor Kokta , Alexendrov Lubo , Sinnett Daniel , Fabre Monique , Vassal Gilles 
Published: March 2012

Sartelet H, Imbriglio T, Nyalendo C, Haddad E, Annabi B, Duval M, Fetni R, Victor K, Alexendrov L, Sinnett D, Fabre M & Vassal G  (2012) Histopathology 60, 1144–1155CD133 expression is associated with poor outcome in neuroblastoma via chemoresistance mediated by the AKT pathwayAims:  Neuroblastoma is a frequent childhood cancer with a heterogeneous prognosis. CD133 expression is an independent prognostic marker for a low survival rate in several cancers. The aim of this study was to determine the prognostic value of CD133 expression in a large cohort of neuroblastoma cases, to define the chemoresistance of neuroblastoma cells expressing CD133, and to determine whether this chemoresistance is regulated by activation of the AKT pathway.Methods and results:  Two hundred and eighty samples of neuroblastoma were screened for CD133 expression. The sensitivity of purified CD133+ neuroblastoma cells isolated from two human cell lines to doxorubicin, vincristine and cisplatin, as single agents or in combination with LY294002, an AKT inhibitor, was evaluated in vitro. CD133 expression was found in 100 of 280 tumours. There was a significant association between CD133 expression and the following poor prognosis covariates: age, International Neuroblastoma Staging System stage, MYCN amplification, and phospho‐AKT (pAKT) expression. Patients with CD133− tumours had significantly better 3‐year event‐free and overall survival than patients with CD133+ tumours. In a multivariate model, CD133 expression was independently associated with decreased overall survival. CD133high neuroblastoma cells were significantly resistant to chemotherapy as compared with CD133low cells. Treatment of unsorted neuroblastoma cells with the three anticancer drugs significantly enriched the CD133+ subpopulation. CD133high cells expressed significantly higher levels of pAKT than CD133low cells. LY294002 treatment abolished the preferential survival of CD133high cells.Conclusions:  CD133 is associated with in‐vitro resistance to chemotherapy involving activation of the AKT pathway.
Immunohistochemical analysis of the endoribonucleases Drosha, Dicer and Ago2 in smooth muscle tumours of soft tissues
Authors: Papachristou Dionysios J , Sklirou Emilia , Corradi Domenico , Grassani Chiara , Kontogeorgakos Vasileios , Rao Uma N M 
Published: March 2012

Papachristou D J, Sklirou E, Corradi D, Grassani C, Kontogeorgakos V & Rao U N M(2012) Histopathology 60, E28–E36Immunohistochemical analysis of the endoribonucleases Drosha, Dicer and Ago2 in smooth muscle tumours of soft tissuesAims:  The aims of the present study were to determine the protein levels and cellular distribution of the endoribonucleases Drosha, Dicer and Ago2, major components of the microRNA‐processing machinery, in benign and malignant soft tissue smooth muscle tumours, and to correlate the cellular levels of these enzymes with clinicopathological parameters including tumour histopathological grade.Methods and results:  Cellular levels of Drosha, Dicer and Ago2 were evaluated in 110 soft tissue leiomyosarcomas (LMS), 31 leiomyomas (LM) and normal smooth muscle (NSM) using immunohistochemistry. Drosha and Dicer were barely detectable in NSM, while augmented levels of these enzymes were observed in LM and LMS. This finding suggests that Drosha and Dicer are implicated in the development of smooth muscle neoplasms. Notably, cellular levels of Dicer were significantly greater in high‐grade compared to low‐grade LMS, implying its participation in the progression of these neoplasms. Ago2 was detected in NSM, as well as LM and LMS; its cellular levels were not associated with tumour grade.Conclusions:  Our results provide novel evidence that Drosha, Dicer and Ago2 are probably involved in the pathobiology of human smooth muscle neoplasms and that Dicer could serve as potentially significant biomarker for LMS progression.
Endodontic management of mandibular central incisor fused to a supernumerary tooth associated with a talon cusp: a case report
Authors: Sachdeva G. S. , Malhotra D. , Sachdeva L. T. , Sharma N. , Negi A. 
Published: March 2012

Sachdeva GS, Malhotra D, Sachdeva LT, Sharma N, Negi A. Endodontic management of mandibular central incisor fused to a supernumerary tooth associated with a talon cusp: a case report. International Endodontic Journal, 45, 590–596, 2012.AbstractAim  To report the successful non‐surgical endodontic management of a mandibular central incisor fused to a supernumerary tooth associated with a talon cusp.Summary  Fusion and gemination are developmental anomalies of teeth that may require endodontic treatment. In this article, a case of successful endodontic management of a permanent mandibular right central incisor fused to its supernumerary counterpart associated with a talon cusp is reported. The incidence of fusion in mandibular anteriors is rare. A search of the literature failed to reveal any reports on the fusion of a permanent mandibular central incisor with its supernumerary counterpart associated with a talon cusp. Successful non‐surgical endodontic management of a case is reported.Key learning points
Fused and geminated teeth requiring endodontic treatment present diagnostic and technical challenges.
An exact differentiation between fusion and gemination may not be critically important for treatment.
The use of an operating microscope for detection of additional root canal orifices in complicated cases is recommended.
Ten‐year follow‐up observations of periapical and endodontic status in a Danish population
Authors: Kirkevang L.‐L. , Væth M. , Wenzel A. 
Published: March 2012

Kirkevang L‐L, Væth M, Wenzel A. 10‐year follow‐up observations of periapical and endodontic status in a Danish population. International Endodontic Journal, 45, 829–839, 2012.AbstractAim  To describe changes in periapical and endodontic status in a general adult Danish population examined in 1997, 2003 and 2008.Methodology  The study provides longitudinal, observational information on endodontic status for 327 individuals who participated in three consecutive full‐mouth radiographic examinations performed with an approximately 5‐year interval. The collected data allow changes to be investigated within and between age‐cohorts.Results  The median number of teeth declined slightly, but the number of individuals with apical periodontitis (AP) and with root filled teeth increased. The number of teeth with AP and of teeth with root fillings increased, whereas the number of root filled teeth with AP was almost stable.The proportion of individuals who had no AP and the proportion of individuals who had no root fillings decreased during the 10‐year observation period.The proportion of teeth with AP/individual increased steadily with age, unaffected by the different time periods. Differences were found between the age groups for the proportion of root filled teeth/individual where the younger age‐cohorts had root fillings performed later in life and at a slower rate. In the old age‐cohort, the proportion of lost teeth/individual was much larger and showed a stronger increase with age, than in the younger age‐cohorts.Conclusion  The younger age‐cohorts will probably never reach the proportion of root fillings or lost teeth seen in the oldest age‐cohort. However, it is disturbing that the proportion of teeth with AP/individual increased steadily with age, completely unaffected by the different time periods.
Serum from patients with erectile dysfunction inhibits circulating angiogenic cells from healthy men: relationship with cardiovascular risk, endothelial damage and circulating angiogenic modulators
Authors: Pelliccione F. , D’Angeli A. , Filipponi S. , Falone S. , Necozione S. , Barbonetti A. , Francavilla F. , Francavilla S. 
Published: March 2012

SummaryErectile dysfunction (ED) is an early manifestation of arteriosclerosis associated with endothelial damage/dysfunction and to a blunted ability of cultured mononuclear circulating cells (MNCs) to differentiate circulating angiogenic cells (CACs), putatively involved in endothelial damage repair. Here we explored effects of human serum (HS) from patients with ED and cardiovascular risk factors (VRFs) but no clinical atherosclerosis, on cultured MNCs of healthy men to differentiate CACs and to form colonies. Effect of HS on number of CACS and of colony forming units (CFUs) was correlated with circulating markers of endothelial damage and with angiogenic modulators. MNCs from healthy men were cultured in standard conditions or with 20% HS from 35 patients with ED and from 10 healthy men. CACs were identified after 7 days of culture by uptake of acetylated low‐density lipoprotein with concomitant binding of Ulex europaeus agglutinin I. CFUs were counted after 5 days of culture. Enzyme‐linked immunosorbent assays assessed plasmatic soluble (s) form of E‐selectin, Endothelin (ET)‐1, tissue type plasminogen activator (tPA), vascular endothelial growth factor (VEGF)165 and sVEGF receptor (R)‐1. The number of CACs and of CFUs from healthy men was reduced after culturing MNCs with HS compared to standard medium. The inhibitory effect was significantly higher with HS from ED patients with higher or lower VRF exposure compared to healthy men. Inhibition was positively correlated with VRFs exposure, with ED severity, with common carotid artery intima media thickness measured using B‐mode ultrasound, and to a lesser extent with plasmatic sE‐Selectin, sET‐1 and sVEGFR‐1. Dysfunction of cells involved in vascular homoeostasis is induced by soluble factors still unknown and already present in a very initial systemic vascular disease in men with ED and VRFs.
No effects of n−3 fatty acid supplementation on serum total testosterone levels in older men: the Alpha Omega Trial
Authors: Giltay E. J. , Geleijnse J. M. , Heijboer A. C. , de Goede J. , Oude Griep L. M. , Blankenstein M. A. , Kromhout D. 
Published: March 2012

SummaryThe intake of the n−3 fatty acids alpha‐linolenic acid (ALA), acid (EPA) and docosahexaenoic acid (DHA) has been related to testosterone levels in epidemiological analyses. The aim of this study was to assess whether the n−3 fatty acids affects testosterone levels in post‐myocardial infarction (MI) patients, who are at risk of testosterone deficiency. In a double‐blind, placebo‐controlled trial of low‐dose supplementation of n−3 fatty acids, we included 1850 male post‐MI patients aged 60–80 years who participated in the Alpha Omega Trial. Patients were randomly allocated to margarines that provided 400 mg/day of EPA–DHA (n = 453), 2 mg/day of ALA (n = 467), EPA–DHA plus ALA (n = 458), or placebo (n = 472). Serum testosterone levels were assessed at baseline and after 41 months using whole day blood samples obtained at the subjects’ home or at the hospital. Subjects were on average age of 68.4 (SD 5.3) years old and had baseline mean serum total testosterone of 14.8 (SD 5.6) nmol/L. The four randomized groups did not differ for baseline characteristics. ALA, EPA–DHA, and EPA–DHA plus ALA supplementation did not affect serum total testosterone compared to placebo. Moreover, n−3 fatty acid supplementation did not affect the risk of incident testosterone deficiency (n = 76 with total testosterone <8.0 nmol/L). We conclude that n−3 fatty acids supplementation did not affect serum total testosterone in men who had had a MI.
Development, reproducibility and validity of the food frequency questionnaire in the Poland arm of the Prospective Urban and Rural Epidemiological (PURE) study
Authors: Dehghan M. , Ilow R. , Zatonska K. , Szuba A. , Zhang X. , Mente A. , Regulska‐Ilow B. 
Published: March 2012

How to cite this article: Dehghan M., Ilow R., Zatonska K., Szuba A., Zhang X., Mente A. & Regulska‐Ilow B. (2012) Development, reproducibility and validity of the food frequency questionnaire in the Poland arm of the Prospective Urban and Rural Epidemiological (PURE) study. J Hum Nutr Diet. 25, 225–232AbstractBackground:  A food frequency questionnaire (FFQ) is the most commonly used method in large epidemiological studies. The validation of an FFQ is essential for specific populations because foods are culture‐dependent. The present study aimed to develop an FFQ and evaluate its validity and reproducibility in estimating the intake of nutrients in urban and rural areas of Poland.Methods:  Adult participants (n = 146) in the Polish arm of the ongoing Prospective Urban and Rural Epidemiological (PURE) study completed FFQs on two occasions, as well as four 24‐h dietary recalls (DRs) during a 12‐month period. Correlation coefficients (r) and de‐attenuated correlation coefficients between dietary recalls and both FFQs were calculated for selected macro‐ and micronutrients. Agreement between the two methods was evaluated by classification into quartiles and the Bland–Altman method. Reproducibility was assessed by the intra‐class correlation coefficient (ICC).Results:  The final food list contained 134 food items. For urban participants, FFQ2 generally underestimated energy, protein and fat compared to the FFQ1 and mean of DRs. In rural areas, compared to DRs, both FFQs overestimated energy and macronutrients. For both urban and rural settings, de‐attenuated correlation exceeded 0.4 for almost all nutrients and the exact agreement in quartile categorisation was >66%. When assessing repeatability, ICC varied from 0.39–0.63 in an urban setting and 0.19–0.45 in a rural setting.Conclusions:  This 134‐item FFQ has good validity and reproducibility in relation to the reference method and can be used to rank individuals based on their macro‐ and micronutrient intake.
Towards deciphering dynamic changes and evolutionary mechanisms involved in the adaptation to low salinities in Ectocarpus (brown algae)
Authors: Dittami Simon M. , Gravot Antoine , Goulitquer Sophie , Rousvoal Sylvie , Peters Akira F. , Bouchereau Alain , Boyen Catherine , Tonon Thierry 
Published: March 2012

SummaryColonizations of freshwater by marine species are rare events, and little information is known about the underlying mechanisms. Brown algae are an independent lineage of photosynthetic and multicellular organisms from which few species inhabit freshwater. As a marine alga that is also found in freshwater, Ectocarpus is of particular interest for studying the transition between these habitats. To gain insights into mechanisms of the transition, we examined salinity tolerance and adaptations to low salinities in a freshwater strain of Ectocarpus on physiological and molecular levels. We show that this isolate belongs to a widely distributed and highly stress‐resistant clade, and differed from the genome‐sequenced marine strain in its tolerance of low salinities. It also exhibited profound, but reversible, morphological, physiological, and transcriptomic changes when transferred to seawater. Although gene expression profiles were similar in both strains under identical conditions, metabolite and ion profiles differed strongly, the freshwater strain exhibiting e.g. higher cellular contents of amino acids and nitrate, higher contents of n‐3 fatty acids, and lower intracellular mannitol and sodium concentrations. Moreover, several stress markers were noted in the freshwater isolate in seawater. This finding suggests that, while high stress tolerance and plasticity may be prerequisites for the colonization of freshwater, genomic alterations have occurred that produced permanent changes in the metabolite profiles to stabilize the transition.
Detection of resistant chickpea (Cicer arietinum) genotypes to Cuscuta campestris (field dodder)
Authors: GOLDWASSER Y , MIRYAMCHIK H , SIBONY M , RUBIN B 
Published: March 2012

Goldwasser Y, Miryamchik H, Sibony M & Rubin B (2012). Detection of resistant chickpea (Cicer arietinum) genotypes to Cuscuta campestris (field dodder). Weed Research 52, 122–130.SummaryThe parasitic plant Cuscuta campestris (field dodder) is the most damaging dodder species, parasitising shoots of broad‐leaved crops. Cuscuta campestris control is extremely difficult because of its hard‐coated seeds, its nature of attachment and intimate host–parasite association. Chickpea is highly susceptible to C. campestris. However, there are few efficient means of selectively managing this parasitic plant in this crop. The aim of this study was to examine and detect the response of chickpea genotypes that are resistant to C. campestris, to identify, understand and utilise them to combat C. campestris. A large‐scale glasshouse screening was conducted to test the response of chickpea genotypes to C. campestris. Next, we tested outstanding resistant genotypes and Israeli varieties. Finally, we confirmed the response of the outstanding resistant genotypes and tested the resistance to secondary dodder parasitism. Most chickpea genotypes tested were susceptible to C. campestris. However, the chickpea genotypes ‘ICCV 95333’ and ‘Hazera 4’ exhibited high resistance; on more than 80% of the chickpea plants, there was no C. campestris development. The resistance phenomenon was characterised by the repelling of the parasite haustoria by the resistant host plant, thus leading to starvation and death of the obligate parasite. Further studies are in progress to elucidate the resistance phenomena and to confirm the resistance under field conditions.
Individual budgets for people with incontinence: results from a ‘shopping’ experiment within the British National Health Service
Authors: Fader Mandy J. , Cottenden Alan M. , Gage Heather M. , Williams Peter , Getliffe Katharine , Clarke‐O’Neill Sinead , Jamieson Katharine M. , Green Nicholas J. 
Published: March 2012

AbstractBackground and context  Most people with urinary incontinence are given limited choice when provided with absorbent products through the British National Health Service (NHS), even though the available range is large.Objective  To investigate users’ preferences for four disposable designs (inserts, all‐in‐ones, belted/T‐shaped and pull‐ups) and towelling washable/reusable products, day and night.Design  Shopping experiment.Setting and participants  Community‐dwelling women and men in England with moderate‐to‐heavy urinary incontinence recruited to a larger trial.Intervention  Participants tested each design and selected products they would prefer with a range of different budgets.Main outcome measures  Design preferences (rankings); ‘purchasing’ decisions from designated budgets.Results  Eighty‐five participants (49 men) tested products, 75 completed the shopping experiment. Inserts, most frequently supplied by the NHS, were ranked second to pull‐ups by women and lowest by men. When faced with budget constraints, up to 40% of participants opted to ‘mix‐and‐match’ designs. Over 15 different combinations of products were selected by participants in the shopping experiment. Most (91%) stated a willingness to ‘top‐up’ assigned budgets from income to secure preferred designs.Discussion  Participants displayed diverse preferences. Enabling user choice of absorbent product design through individual budgets could improve satisfaction of consumers and efficiency of allocation of limited NHS resources.Conclusion  Recent policy for the NHS seeks to provide consumers with more control in their care. Extension of the concept of individual budgets to continence supplies could be feasible and beneficial for patients and provide better value‐for‐money within the NHS. Further research is warranted.
Beyond needs and expectations: identifying the barriers and facilitators to written medicine information provision and use in Australia
Authors: Hamrosi Kim K. , Aslani Parisa , Raynor David K. 
Published: March 2012

AbstractPurpose This study aimed to explore peoples’ needs and expectations of written medicines information (WMI), and to determine the barriers and facilitators experienced or perceived in the context of WMI provision and use.Methods We conducted eight focus groups with 62 participants over 6 weeks in late 2008 in New South Wales, Australia. Using a semi‐structured topic schedule and examples of WMI from Australia and other English‐speaking countries as a guide, we explored themes relevant to WMI, including participant experiences, attitudes, beliefs and expectations.Findings Our findings suggest less than half had previously received WMI, with many unaware of its availability. Many, but not all, wanted WMI to supplement the spoken information they received but not to replace it, and it was predominantly used to facilitate informed choice, ascertain medicine suitability and review instructions. The current leaflets were considered technical and long, and a summary leaflet in addition to comprehensive information was favoured. Accurate side‐effect information was the most important element that participants desired. The most common barriers to effective WMI use were time constraints and patient confidence, with participants citing empowerment, time and health‐care professional (HCP)–patient relationships as important facilitators.Conclusion The findings provide insight and understanding of peoples needs and expectations, and clarify issues associated with use and non‐use of WMI. Challenges include addressing the barriers, especially of time and HCP attitudes to drive changes to workplace practices, and learning from the facilitating factors to encourage awareness and accessibility to WMI as a tool to empower patients.