bluuebunny/crossref_metadata_2025 huggingface dataset:
The migration of paraxial and lateral plate mesoderm cells emerging from the late primitive streak is controlled by different Wnt signals
Authors: Sweetman Dylan , Wagstaff Laura , Cooper Oliver , Weijer Cornelis , Münsterberg Andrea
Published: June 2008

Background

Co-ordinated cell movement is a fundamental feature of developing embryos. Massive cell movements occur during vertebrate gastrulation and during the subsequent extension of the embryonic body axis. These are controlled by cell-cell signalling and a number of pathways have been implicated. Here we use long-term video microscopy in chicken embryos to visualize the migration routes and movement behaviour of mesoderm progenitor cells as they emerge from the primitive streak (PS) between HH stages 7 and 10.

Results

We observed distinct cell movement behaviours along the length of the streak and determined that this is position dependent with cells responding to environmental cues. The behaviour of cells was altered by exposing embryos or primitive streak explants to cell pellets expressing Wnt3a and Wnt5a, without affecting cell fates, thus implicating these ligands in the regulation of cell movement behaviour. Interestingly younger embryos were not responsive, suggesting that Wnt3a and Wnt5a are specifically involved in the generation of posterior mesoderm, consistent with existing mouse and zebrafish mutants. To investigate which downstream components are involved mutant forms of dishevelled (dsh) and prickle1 (pk1) were electroporated into the primitive streak. These had differential effects on the behaviour of mesoderm progenitors emerging from anterior or posterior regions of the streak, suggesting that multiple Wnt pathways are involved in controlling cell migration during extension of the body axis in amniote embryos.

Conclusion

We suggest that the distinct behaviours of paraxial and lateral mesoderm precursors are regulated by the opposing actions of Wnt5a and Wnt3a as they leave the primitive streak in neurula stage embryos. Our data suggests that Wnt5a acts via prickle to cause migration of cells from the posterior streak. In the anterior streak, this is antagonised by Wnt3a to generate non-migratory medial mesoderm.
atonal- and achaete-scute-related genes in the annelid Platynereis dumerilii: insights into the evolution of neural basic-Helix-Loop-Helix genes
Authors: Simionato Elena , Kerner Pierre , Dray Nicolas , Le Gouar Martine , Ledent Valérie , Arendt Detlev , Vervoort Michel
Published: June 2008

Background

Functional studies in model organisms, such as vertebrates andDrosophila, have shown that basic Helix-loop-Helix (bHLH) proteins have important roles in different steps of neurogenesis, from the acquisition of neural fate to the differentiation into specific neural cell types. However, these studies highlighted many differences in the expression and function of orthologous bHLH proteins during neural development between vertebrates andDrosophila. To understand how the functions of neural bHLH genes have evolved among bilaterians, we have performed a detailed study of bHLH genes during nervous system development in the polychaete annelid,Platynereis dumerilii, an organism which is evolutionary distant from bothDrosophilaand vertebrates.

Results

We have studiedPlatynereisorthologs of the most important vertebrate neural bHLH genes, i.e.achaete-scute, neurogenin, atonal, olig, andNeuroDgenes, the latter two being genes absent of theDrosophilagenome. We observed that all these genes have specific expression patterns during nervous system formation inPlatynereis. Our data suggest that inPlatynereis, like in vertebrates but unlikeDrosophila, (i)neurogeninis the main proneural gene for the formation of the trunk central nervous system, (ii)achaete-scuteandoliggenes are involved in neural subtype specification in the central nervous system, in particular in the specification of the serotonergic phenotype. In addition, we found that thePlatynereis NeuroDgene has a broad and early neuroectodermal expression, which is completely different from the neuronal expression of vertebrateNeuroDgenes.

Conclusion

Our analysis suggests that thePlatynereisbHLH genes have both proneural and neuronal specification functions, in a way more akin to the vertebrate situation than to that ofDrosophila. We conclude that these features are ancestral to bilaterians and have been conserved in the vertebrates and annelids lineages, but have diverged in the evolutionary lineage leading toDrosophila.
A simple and accurate SNP scoring strategy based on typeIIS restriction endonuclease cleavage and matrix-assisted laser desorption/ionization mass spectrometry
Authors: Hong Sun Pyo , Ji Seung Il , Rhee Hwanseok , Shin Soo Kyeong , Hwang Sun Young , Lee Seung Hwan , Lee Soong Deok , Oh Heung-Bum , Yoo Wangdon , Kim Soo-Ok
Published: June 2008

Background

We describe the development of a novel matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)-based single nucleotide polymorphism (SNP) scoring strategy, termed Restriction Fragment Mass Polymorphism (RFMP) that is suitable for genotyping variations in a simple, accurate, and high-throughput manner. The assay is based on polymerase chain reaction (PCR) amplification and mass measurement of oligonucleotides containing a polymorphic base, to which a typeIIS restriction endonuclease recognition was introduced by PCR amplification. Enzymatic cleavage of the products leads to excision of oligonucleotide fragments representing base variation of the polymorphic site whose masses were determined by MALDI-TOF MS.

Results

The assay represents an improvement over previous methods because it relies on the direct mass determination of PCR products rather than on an indirect analysis, where a base-extended or fluorescent report tag is interpreted. The RFMP strategy is simple and straightforward, requiring one restriction digestion reaction following target amplification in a single vessel. With this technology, genotypes are generated with a high call rate (99.6%) and high accuracy (99.8%) as determined by independent sequencing.

Conclusion

The simplicity, accuracy and amenability to high-throughput screening analysis should make the RFMP assay suitable for large-scale genotype association study as well as clinical genotyping in laboratories.
CRISTOPH – A cluster-randomised intervention study to optimise the treatment of patients with hypertension in General Practice
Authors: Mortsiefer Achim , Meysen Tobias , Schumacher Martin , Lintges Claudia , Stamer Maren , Schmacke Norbert , Wegscheider Karl , Abholz Heinz-Harald , in der Schmitten Jürgen
Published: June 2008

Background

Recent guidelines for the management of hypertension focus on treating patients according to their global cardiovascular risk (CVR), rather than strictly keeping blood pressure, or other risk factors, below set limit values. The objective of this study is to compare the effect of a simple versus a complex educational intervention implementing this new concept among General Practitioners (GPs).

Methods/design

A prospective longitudinal cluster-randomised intervention trial with 94 German GPs consecutively enroling 40 patients each with known hypertension. All GPs then received a written manual specifically developed to transfer the global concept of CVR into daily General Practice. After cluster-randomisation, half of the GPs additionally received a clinical outreach visit, with a trained peer discussing with them the concept of global CVR referring to example study patients from the respective GP. Main outcome measure is the improvement of calculated CVR six months after intervention in the subgroup of patients with high CVR (but no history of cardiovascular disease), defined as 10-year-mortality ≥ 5% employing the European SCORE formula. Secondary outcome measures include the intervention's effect on single risk factors, and on prescription rates of drugs targeting CVR. All outcome measures are separately studied in the three subgroups of patients with 1. high CVR (defined as above), 2. low CVR (SCORE < 5%), and 3. a history of cardiovascular disease. The influence of age, sex, social status, and the perceived quality of the respective doctor-patient-relation on the effects will be examined.

Discussion

To our knowledge, no other published intervention study has yet evaluated the impact of educating GPs with the goal to treat patients with hypertension according to their global cardiovascular risk.

Trial registration

ISRCTN44478543
Utility of 18F-fluoro-deoxyglucose emission tomography/computed tomography fusion imaging (18F-FDG PET/CT) in combination with ultrasonography for axillary staging in primary breast cancer
Authors: Ueda Shigeto , Tsuda Hitoshi , Asakawa Hideki , Omata Jiro , Fukatsu Kazuhiko , Kondo Nobuo , Kondo Tadaharu , Hama Yukihiro , Tamura Katsumi , Ishida Jiro , Abe Yoshiyuki , Mochizuki Hidetaka
Published: June 2008

Background

Accurate evaluation of axillary lymph node (ALN) involvement is mandatory before treatment of primary breast cancer. The aim of this study is to compare preoperative diagnostic accuracy between positron emission tomography/computed tomography with 18F-fluorodeoxyglucose (18F-FDG PET/CT) and axillary ultrasonography (AUS) for detecting ALN metastasis in patients having operable breast cancer, and to assess the clinical management of axillary 18F-FDG PET/CT for therapeutic indication of sentinel node biopsy (SNB) and preoperative systemic chemotherapy (PSC).

Methods

One hundred eighty-three patients with primary operable breast cancer were recruited. All patients underwent 18F-FDG PET/CT and AUS followed by SNB and/or ALN dissection (ALND). Using 18F-FDG PET/CT, we studied both a visual assessment of 18F-FDG uptake and standardized uptake value (SUV) for axillary staging.

Results

In a visual assessment of 18F-FDG PET/CT, the diagnostic accuracy of ALN metastasis was 83% with 58% in sensitivity and 95% in specificity, and when cut-off point of SUV was set at 1.8, sensitivity, specificity, and accuracy were 36, 100, and 79%, respectively. On the other hand, the diagnostic accuracy of AUS was 85% with 54% in sensitivity and 99% in specificity. By the combination of 18F-FDG PET/CT and AUS to the axilla, the sensitivity, specificity, and accuracy were 64, 94, and 85%, respectively. If either 18F-FDG PET uptake or AUS was positive in allixa, the probability of axillary metastasis was high; 50% (6 of 12) in 18F-FDG PET uptake only, 80% (4 of 5) in AUS positive only, and 100% (28 of 28) in dual positive. By the combination of AUS and 18F-FDG PET/CT, candidates of SNB were more appropriately selected. The axillary 18F-FDG uptake was correlated with the maximum size and nuclear grade of metastatic foci (p = 0.006 and p = 0.03).

Conclusion

The diagnostic accuracy of 18F-FDG PET/CT was shown to be nearly equal to ultrasound, and considering their limited sensitivities, the high radiation exposure by 18F-FDG PET/CT and also costs of the examination, it is likely that AUS will be more cost-effective in detecting massive axillary tumor burden. However, when we cannot judge the axillary staging using AUS alone, metabolic approach of 18F-FDG PET/CT for axillary staging would enable us a much more confident diagnosis.
Intravenous postoperative fluid prescriptions for children: A survey of practice
Authors: Davies Polly , Hall Tim , Ali Tariq , Lakhoo Kokila
Published: June 2008

Background

Postoperative deaths and neurological injury have resulted from hyponatraemia associated with the use of hypotonic saline solutions following surgery. We aimed to determine the rates and types of intravenous fluids being prescribed postoperatively for children in the UK.

Methods

A questionnaire was sent to members of the British Association of Paediatric Surgeons (BAPS) and Association of Paediatric Anaesthetists of Great Britain and Ireland (APAGBI) based at UK paediatric centres. Respondents were asked to prescribe postoperative fluids for scenarios involving children of different ages. The study period was between May 2006 and November 2006.

Results

The most frequently used solution was sodium chloride 0.45% with glucose 5% although one quarter of respondents still used sodium chloride 0.18% with glucose 4%. Isotonic fluids were used by 41% of anaesthetists and 9.8% of surgeons for the older child, but fewer for infants. Standard maintenance rates or greater were prescribed by over 80% of respondents.

Conclusion

Most doctors said they would prescribe hypotonic fluids at volumes equal to or greater than traditional maintenance rates at the time of the survey. A survey to describe practice since publication of National Patient Safety Agency (NPSA) recommendations is required.
ret/PTC-1 expression alters the immunoprofile of thyroid follicular cells
Authors: Denning Karen , Smyth Paul , Cahill Susanne , Li Jinghuan , Flavin Richard , Aherne Sinead , O' Leary John J , Sheils Orla
Published: June 2008

Background

Hashimoto Thyroiditis (H.T.) is a destructive autoimmune thyroid condition whose precise molecular pathogenesis remains unclear. ret/PTC-1 is a chimeric transcript which has been described in autoimmune thyroid disease (AITD) and thyroid neoplasia. The purpose of this study was to observe the immunogenic effect exposure to H.T. and control lymphocyte supernatant would have on normal (Nthy-ori) and ret/PTC-1 (TPC-1) expressing thyroid cell line models.

Results

A 2 × 2 matrix comprising Nthy-ori and TPC-1 cell lines and H.T. and control lymphocyte supernatant was designed and utilised as follows; activated lymphocytic supernatant from a H.T. and normal control were co-cultured with a cell line derived from normal thyroid (Nthy-ori) and also a cell line derived from a papillary thyroid carcinoma that endogenously expresses ret/PTC-1 (TPC-1). The co-cultures were harvested at 0, 6 and 18 hour time points. Gene expression analysis was performed on RNA extracted from thyrocytes using TaqMan® Immune profiling Low-Density Arrays (Applied Biosystems, CA, USA) comprising gene expression markers for 93 immune related targets plus 3 endogenous controls.

Stimulation of the normal thyroid cell line model with activated T cell supernatant from the H.T. donor yielded global up-regulation of immune targets when compared with control supernatant stimulation. In particular, a cohort of targets (granzyme B, CD3, CD25, CD152, CD45) associated with cytotoxic cell death; T cell receptor (TCR) and T cell signaling were up-regulated in the normal cell line model. When the ret/PTC-1 expressing thyroid cell line was co-cultured with H.T. lymphocyte supernatant, in comparison to control supernatant stimulation, down-regulation of the same subset of immune targets was seen.

Conclusion

Co-culturing H.T. lymphocyte supernatant with a normal thyroid cell line model leads to over-expression of a subset of targets which could contribute to the pathogenesis of H.T. via cytotoxic cell death and TCR signalling. Stimulation of the ret/PTC-1 positive cell line with the same stimulus led to a down-regulated shift in the gene expression pattern of the cohort of immune targets. We hypothesize that ret/PTC-1 activation may dampen immunogenic responses in the thyroid, which could possibly facilitate papillary thyroid carcinoma development.
Stabilizing mutation of CTNNB1/beta-catenin and protein accumulation analyzed in a large series of parathyroid tumors of Swedish patients
Authors: Björklund Peyman , Lindberg Daniel , Åkerström Göran , Westin Gunnar
Published: June 2008

Background

Aberrant accumulation of β-catenin plays an important role in a variety of human neoplasms. We recently reported accumulation of β-catenin in parathyroid adenomas from patients with primary hyperparathyroidism (pHPT). In CTNNB1 exon 3, we detected a stabilizing mutation (S37A) in 3 out of 20 analyzed adenomas. The aim of the present study was to determine the frequency and zygosity of mutations in CTNNB1 exon 3, and β-catenin accumulation in a large series of parathyroid adenomas of Swedish patients.

Results

The mutation S37A (TCT > GCT) was detected by direct DNA sequencing of PCR fragments in 6 out of 104 sporadic parathyroid adenomas (5.8%). Taking our previous study into account, a total of 9 out of 124 (7.3%) adenomas displayed the same mutation. The mutations were homozygous by DNA sequencing, restriction enzyme cleavage, and gene copy number determination using the GeneChip 500 K Mapping Array Set. All tumors analyzed by immunohistochemistry, including those with mutation, displayed aberrant β-catenin accumulation. Western blotting revealed a slightly higher expression level of β-catenin and nonphosphorylated active β-catenin in tumors with mutation compared to those without. Presence of the mutation was not related to distinct clinical characteristics.

Conclusion

Aberrant accumulation of β-catenin is very common in parathyroid tumors, and is caused by stabilizing homozygous mutation in 7.3% of Swedish pHPT patients.
Nutritional skewing of conceptus sex in sheep: effects of a maternal diet enriched in rumen-protected polyunsaturated fatty acids (PUFA)
Authors: Green Mark P , Spate Lee D , Parks Tina E , Kimura Koji , Murphy Clifton N , Williams Jim E , Kerley Monty S , Green Jonathan A , Keisler Duane H , Roberts R Michael
Published: June 2008

Background

Evolutionary theory suggests that in polygynous mammalian species females in better body condition should produce more sons than daughters. Few controlled studies have however tested this hypothesis and controversy exists as to whether body condition score or maternal diet is in fact the determining factor of offspring sex. Here, we examined whether maternal diet, specifically increased n-6 polyunsaturated fatty acid (PUFA) intake, of ewes with a constant body condition score around the time of conception influenced sex ratio.

Methods

Ewes (n = 44) maintained in similar body condition throughout the study were assigned either a control (C) diet or one (F) enriched in rumen-protected PUFA, but otherwise essentially equivalent, from four weeks prior to breeding until d13 post-estrus. On d13, conceptuses were recovered, measured, cultured to assess their capacity for interferon-tau (IFNT) production and their sex determined. The experiment was repeated with all ewes being fed the F diet to remove any effects of parity order on sex ratio. Maternal body condition score (BCS), plasma hormone and metabolite concentrations were also assessed throughout the study and related to diet.

Results

In total 129 conceptuses were recovered. Ewes on the F diet produced significantly more male than female conceptuses (proportion male = 0.69; deviation from expected ratio of 0.5, P < 0.001). Conceptus IFNT production was unaffected by diet (P > 0.1), but positively correlated with maternal body condition score (P < 0.05), and was higher (P < 0.05) in female than male conceptuses after 4 h culture. Maternal plasma hormone and metabolite concentrations, especially progesterone and fatty acid, were also modulated by diet.

Conclusion

These results provide evidence that maternal diet, in the form of increased amounts of rumen-protected PUFA fed around conception, rather than maternal body condition, can skew the sex ratio towards males. These observations may have implications to the livestock industry and animal management policies when offspring of one sex may be preferred over the other.
A subgroup of plant aquaporins facilitate the bi-directional diffusion of As(OH)3 and Sb(OH)3across membranes
Authors: Bienert Gerd P , Thorsen Michael , Schüssler Manuela D , Nilsson Henrik R , Wagner Annemarie , Tamás Markus J , Jahn Thomas P
Published: June 2008

Background

Arsenic is a toxic and highly abundant metalloid that endangers human health through drinking water and the food chain. The most common forms of arsenic in the environment are arsenate (As(V)) and arsenite (As(III)). As(V) is a non-functional phosphate analog that enters the food chain via plant phosphate transporters. Inside cells, As(V) becomes reduced to As(III) for subsequent extrusion or compartmentation. Although much is known about As(III) transport and handling in microbes and mammals, the transport systems for As(III) have not yet been characterized in plants.

Results

Here we show that the Nodulin26-like Intrinsic Proteins (NIPs) AtNIP5;1 and AtNIP6;1 from Arabidopsis thaliana, OsNIP2;1 and OsNIP3;2 from Oryza sativa, and LjNIP5;1 and LjNIP6;1 from Lotus japonicus are bi-directional As(III) channels. Expression of these NIPs sensitized yeast cells to As(III) and antimonite (Sb(III)), and direct transport assays confirmed their ability to facilitate As(III) transport across cell membranes. On medium containing As(V), expression of the same NIPs improved yeast growth, probably due to increased As(III) efflux. Our data furthermore provide evidence that NIPs can discriminate between highly similar substrates and that they may have differential preferences in the direction of transport. A subgroup of As(III) permeable channels that group together in a phylogenetic tree required N-terminal truncation for functional expression in yeast.

Conclusion

This is the first molecular identification of plant As(III) transport systems and we propose that metalloid transport through NIPs is a conserved and ancient feature. Our observations are potentially of great importance for improved remediation and tolerance of plants, and may provide a key to the development of low arsenic crops for food production.
Expression of parathyroid hormone-related protein during immortalization of human peripheral blood mononuclear cells by HTLV-1: Implications for transformation
Authors: Nadella Murali VP , Shu Sherry T , Dirksen Wessel P , Thudi Nanda K , Nadella Kiran S , Fernandez Soledad A , Lairmore Michael D , Green Patrick L , Rosol Thomas J
Published: June 2008

Background

Adult T-cell leukemia/lymphoma (ATLL) is initiated by infection with human T-lymphotropic virus type-1 (HTLV-1); however, additional host factors are also required for T-cell transformation and development of ATLL. The HTLV-1 Tax protein plays an important role in the transformation of T-cells although the exact mechanisms remain unclear. Parathyroid hormone-related protein (PTHrP) plays an important role in the pathogenesis of humoral hypercalcemia of malignancy (HHM) that occurs in the majority of ATLL patients. However, PTHrP is also up-regulated in HTLV-1-carriers and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients without hypercalcemia, indicating that PTHrP is expressed before transformation of T-cells. The expression of PTHrP and the PTH/PTHrP receptor during immortalization or transformation of lymphocytes by HTLV-1 has not been investigated.

Results

We report that PTHrP was up-regulated during immortalization of lymphocytes from peripheral blood mononuclear cells by HTLV-1 infection in long-term co-culture assays. There was preferential utilization of the PTHrP-P2 promoter in the immortalized cells compared to the HTLV-1-transformed MT-2 cells. PTHrP expression did not correlate temporally with expression of HTLV-1 tax. HTLV-1 infection up-regulated the PTHrP receptor (PTH1R) in lymphocytes indicating a potential autocrine role for PTHrP. Furthermore, co-transfection of HTLV-1 expression plasmids and PTHrP P2/P3-promoter luciferase reporter plasmids demonstrated that HTLV-1 up-regulated PTHrP expression only mildly, indicating that other cellular factors and/or events are required for the very high PTHrP expression observed in ATLL cells. We also report that macrophage inflammatory protein-1α (MIP-1α), a cellular gene known to play an important role in the pathogenesis of HHM in ATLL patients, was highly expressed during early HTLV-1 infection indicating that, unlike PTHrP, its expression was enhanced due to activation of lymphocytes by HTLV-1 infection.

Conclusion

These data demonstrate that PTHrP and its receptor are up-regulated specifically during immortalization of T-lymphocytes by HTLV-1 infection and may facilitate the transformation process.
HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells
Authors: Lemay Julie , Maidou-Peindara Priscilla , Bader Thomas , Ennifar Eric , Rain Jean-Christophe , Benarous Richard , Liu Lang Xia
Published: June 2008

Reverse transcription of the genetic material of human immunodeficiency virus type 1 (HIV-1) is a critical step in the replication cycle of this virus. This process, catalyzed by reverse transcriptase (RT), is well characterized at the biochemical level. However, in infected cells, reverse transcription occurs in a multiprotein complex – the reverse transcription complex (RTC) – consisting of viral genomic RNA associated with viral proteins (including RT) and, presumably, as yet uncharacterized cellular proteins. Very little is known about the cellular proteins interacting with the RTC, and with reverse transcriptase in particular. We report here that HIV-1 reverse transcription is affected by the levels of a nucleocytoplasmic shuttling protein – the RNA-binding protein HuR. A direct protein-protein interaction between RT and HuR was observed in a yeast two-hybrid screen and confirmed in vitro by homogenous time-resolved fluorescence (HTRF). We mapped the domain interacting with HuR to the RNAse H domain of RT, and the binding domain for RT to the C-terminus of HuR, partially overlapping the third RRM RNA-binding domain of HuR. HuR silencing with specific siRNAs greatly impaired early and late steps of reverse transcription, significantly inhibiting HIV-1 infection. Moreover, by mutagenesis and immunoprecipitation studies, we could not detect the binding of HuR to the viral RNA. These results suggest that HuR may be involved in and may modulate the reverse transcription reaction of HIV-1, by an as yet unknown mechanism involving a protein-protein interaction with HIV-1 RT.
Mammosphere culture of metastatic breast cancer cells enriches for tumorigenic breast cancer cells
Authors: Grimshaw Matthew J , Cooper Lucienne , Papazisis Konstantinos , Coleman Julia A , Bohnenkamp Hermann R , Chiapero-Stanke Laura , Taylor-Papadimitriou Joyce , Burchell Joy M
Published: June 2008

Introduction

The identification of potential breast cancer stem cells is of importance as the characteristics of stem cells suggest that they are resistant to conventional forms of therapy. Several techniques have been proposed to isolate or enrich for tumorigenic breast cancer stem cells, including (a) culture of cells in non-adherent non-differentiating conditions to form mammospheres and (b) sorting of the cells by their surface phenotype (expression of CD24 and CD44).

Methods

We have cultured metastatic cells found in pleural effusions from breast cancer patients in non-adherent conditions without serum to form mammospheres. Dissociated cells from these mammospheres were used to determine the tumorigenicity of these cultures. Expression of CD24 and CD44 on uncultured cells and mammospheres derived from the pleural effusions was documented.

Results

We found that the majority (20/27) of the pleural effusions tested contained cells capable of forming mammospheres of varying sizes that could be passaged. After dissociation and plating with serum onto adherent dishes, the cells can differentiate, as determined by the increased expression of cytokeratins and MUC1. Analysis of surface expression of CD24 and CD44 on uncultured cells from 21 of the samples showed that the cells from some samples separated into two populations, but some did not. The proportion of cells that could be considered CD44+/CD24low/- was highly variable and did not appear to correlate with the ability to form the larger mammospheres. Of eight pleural effusion mammospheres tested in severe combined immunodeficiency disease (SCID) mice, four were found to induce tumours when only 5,000 or fewer cells were injected, whereas the same number of uncultured cells did not form tumours. The ability to induce tumours appeared to correlate with the ability to produce the larger mammospheres. Uncultured cells from a highly tumorigenic sample (PE14) were uniformly negative for surface expression of both CD24 and CD44.

Conclusion

This paper shows, for the first time, that mammosphere culture of pleural effusions enriches for cells capable of inducing tumours in SCID mice. The data suggest that mammosphere culture of these metastatic cells could provide a highly appropriate model for studying the sensitivity of the tumorigenic 'stem' cells to therapeutic agents and for further characterisation of the tumour-inducing subpopulation of breast cancer cells.
Evidence of the Oncologic Superiority of Cylindrical Abdominoperineal Excision for Low Rectal Cancer
Authors: West Nicholas P. , Finan Paul J. , Anderin Claes , Lindholm Johan , Holm Torbjorn , Quirke Philip
Published: June 2008

Purpose

Abdominoperineal excision (APE) of the rectum and anus for rectal cancer continues to have greater local recurrence and poorer survival than that seen following anterior resection. Changing to an extended prone perineal dissection results in a more cylindrical specimen and should improve outcomes.

Patients and Methods

One hundred twenty-eight specimens from patients who underwent APE that was performed for potentially curable primary rectal adenocarcinoma were dissected according to standard protocol in Leeds and Stockholm between 1997 and 2007 and were studied. Tissue morphometry was performed on the cross sectional photographs of 93 patient cases.

Results

The cylindrical technique removed more tissue in the distal rectum and in all slices that contained tumor compared with the standard operation (both P < .0001). Greater distance was observed from the muscularis propria or internal sphincter to the anterior, posterior, and lateral resection margins (all P < .0001). This was associated with lower circumferential resection margin (CRM) involvement (14.8% v 40.6%; P = .013) and intraoperative perforations (3.7% v 22.8%; P = .0255). An increase in the amount of tissue removed in the distal rectum (P < .0001) was demonstrated by a single surgeon who changed from the standard to the cylindrical technique during the study period; the change was associated with a reduction in CRM positivity (from 36.2% to 12.5%) and in perforations (from 12.8% to 0.0%).

Conclusion

Cylindrical APE performed in the prone position for low rectal cancer removes more tissue around the tumor that leads to a reduction in CRM involvement and intraoperative perforations, which should reduce local disease recurrence. The cylindrical technique has the potential to improve patient outcomes substantially if appropriate surgical education programs are developed.
Intermittent Target Inhibition With Dasatinib 100 mg Once Daily Preserves Efficacy and Improves Tolerability in Imatinib-Resistant and -Intolerant Chronic-Phase Chronic Myeloid Leukemia
Authors: Shah Neil P. , Kantarjian Hagop M. , Kim Dong-Wook , Réa Delphine , Dorlhiac-Llacer Pedro E. , Milone Jorge H. , Vela-Ojeda Jorge , Silver Richard T. , Khoury H. Jean , Charbonnier Aude , Khoroshko Nina , Paquette Ronald L. , Deininger Michael , Collins Robert H. , Otero Irma , Hughes Timothy , Bleickardt Eric , Strauss Lewis , Francis Stephen , Hochhaus Andreas
Published: June 2008

Purpose

Dasatinib is a BCR-ABL inhibitor, 325-fold more potent than imatinib against unmutated BCR-ABL in vitro. Phase II studies have demonstrated efficacy and safety with dasatinib 70 mg twice daily in chronic-phase (CP) chronic myelogenous leukemia (CML) after imatinib treatment failure. In phase I, responses occurred with once-daily administration despite only intermittent BCR-ABL inhibition. Once-daily treatment resulted in less toxicity, suggesting that toxicity results from continuous inhibition of unintended targets. Here, a dose- and schedule-optimization study is reported.

Patients and Methods

In this open-label phase III trial, 670 patients with imatinib-resistant or -intolerant CP-CML were randomly assigned 1:1:1:1 between four dasatinib treatment groups: 100 mg once daily, 50 mg twice daily, 140 mg once daily, or 70 mg twice daily.

Results

With minimum follow-up of 6 months (median treatment duration, 8 months; range, < 1 to 15 months), marked and comparable hematologic (complete, 86% to 92%) and cytogenetic (major, 54% to 59%; complete, 41% to 45%) response rates were observed across the four groups. Time to and duration of cytogenetic response were similar, as was progression-free survival (8% to 11% of patients experienced disease progression or died). Compared with the approved 70-mg twice-daily regimen, dasatinib 100 mg once daily resulted in significantly lower rates of pleural effusion (all grades, 7% v 16%; P = .024) and grade 3 to 4 thrombocytopenia (22% v 37%; P = .004), and fewer patients required dose interruption (51% v 68%), reduction (30% v 55%), or discontinuation (16% v 23%).

Conclusion

Dasatinib 100 mg once daily retains the efficacy of 70 mg twice daily with less toxicity. Intermittent target inhibition with tyrosine kinase inhibitors may preserve efficacy and reduce adverse events.
Mature and Immature Extracranial Teratomas in Children: The UK Children's Cancer Study Group Experience
Authors: Mann Jillian R. , Gray Elizabeth S. , Thornton Claire , Raafat Faro , Robinson Kathleen , Collins Gary S. , Gornall Peter , Huddart Simon N. , Hale Juliet P. , Oakhill Anthony
Published: June 2008

Purpose

The purpose of this article is to describe the features, treatment, and risk factors for relapse of children with mature teratoma (MT) and immature teratoma (IT) to assist future treatment plans.

Patients and Methods

Patients were younger than 16 years of age and referred to the UK Children's Cancer Study Group centers with biopsy-proven extracranial MT and IT and no prior chemotherapy. Complete excision, with the coccyx in sacrococcygeal patients, and follow-up, including serum alpha-fetoprotein monitoring for early detection of malignant yolk sac tumor (YST) recurrence, were recommended. Carboplatin, etoposide, and bleomycin (JEB) were given for YST relapse, whereas relapsed MT and IT were treated at clinicians’ discretion, usually surgically. Pathology was reviewed and treatments, outcome, and prognostic features assessed.

Results

There were 351 patients, 227 with MT, 124 with IT. Tumor sites were: testis (n = 53), ovary (n = 130), sacrococcygeal region (n = 98), thorax (n = 23), and other (n = 47). Surgical resection was incomplete in 26% of MT and 40% of IT patients; 5-year event-free survival was 92.2% and 85.9%, respectively, and 5-year overall survival was 99% and 95.1%. Poorer outcome occurred with incomplete resection, tumor rupture, nongonadal site (particularly sacrococcygeal), young age, higher stage and grade, and gliomatosis peritonei, but not with cyst fluid aspiration/spillage, tumor enucleation, nodal gliomatosis, or microfoci of YST in the tumor (Heifetz lesions). JEB was effective for YST recurrence, but not for MT or IT.

Conclusion

Treatment remains primarily surgical, with JEB chemotherapy for YST relapse. No definite response followed JEB for pure MT and IT. Adjuvant chemotherapy after surgery for sacrococcygeal patients is not advocated.
The cis-acting replication elements define human enterovirus and rhinovirus species
Authors: Cordey Samuel , Gerlach Daniel , Junier Thomas , Zdobnov Evgeny M. , Kaiser Laurent , Tapparel Caroline
Published: June 2008

Replication of picornaviruses is dependent on VPg uridylylation, which is linked to the presence of the internal cis-acting replication element (cre). Cre are located within the sequence encoding polyprotein, yet at distinct positions as demonstrated for poliovirus and coxsackievirus-B3, cardiovirus, and human rhinovirus (HRV-A and HRV-B), overlapping proteins 2C, VP2, 2A, and VP1, respectively. Here we report a novel distinct cre element located in the VP2 region of the recently reported HRV-A2 species and provide evolutionary evidence of its functionality. We also experimentally interrogated functionality of recently identified HRV-B cre in the 2C region that is orthologous to the human enterovirus (HEV) cre and show that it is dispensable for replication and appears to be a nonfunctional evolutionary relic. In addition, our mutational analysis highlights two amino acids in the 2C protein that are crucial for replication. Remarkably, we conclude that each genetic clade of HRV and HEV is characterized by a unique functional cre element, where evolutionary success of a new genetic lineage seems to be associated with an invention of a novel cre motif and decay of the ancestral one. Therefore, we propose that cre element could be considered as an additional criterion for human rhinovirus and enterovirus classification.
Avoiding stimulus confounds in Implicit Association Tests by using the concepts as stimuli
Authors: Steffens Melanie C. , Kirschbaum Michael , Glados Petra
Published: June 2008

Implicit Association Tests (IATs) are supposed to measure associations between concepts. In order to achieve that aim, participants are required to assign individual stimuli to those concepts under time pressure in two different tasks. Previous research has shown that not only the associations of the concepts with each other, but also the stimuli's cross‐category associations influence the observed reaction time difference between these tasks (i.e. the IAT effect). Little is known about adequate stimulus selection. In this article, we introduce a variant of the IAT, the Concept Association Task (CAT) in which the concepts themselves or synonyms of them are used as stimuli. Three experiments on Germans' attitudes towards foreigners yielded evidence for the convergent validity of the CAT: (1) it correlated well with other IAT versions; (2) it correlated higher with spontaneous attitude‐related judgements than other IAT versions; and (3) it correlated with response‐window priming, another implicit measure based on reaction times. Furthermore, we showed that the CAT yielded reasonable findings when other IAT versions appear to yield distorted ones.
Pregnancy Intention and Preterm Birth: Differential Associations Among a Diverse Population of Women
Authors: Afable‐Munsuz Aimee , Braveman Paula
Published: June 2008

CONTEXT:  Studies published to date provide mixed evidence on the relationship between unintended pregnancy and preterm birth, and none take into consideration that the meaning of unintended pregnancy may vary across racial and ethnic groups.

METHODS:  Data from the 1999–2003 rounds of the Maternal and Infant Health Assessment, a population‐based, representative survey of postpartum women in California, were used to assess the relationship between pregnancy intention and preterm birth. For racial and ethnic groups in which an association was found, sequential logistic regression was conducted to further examine the relationship while controlling for socioeconomic characteristics.

RESULTS:  In unadjusted results, pregnancy intention was associated with preterm birth among both whites and immigrant Latinas, but not among blacks or U.S.‐born Latinas. Among whites, compared with women who reported that their pregnancy was intended, those who were unsure about their pregnancy had elevated odds of preterm birth (odds ratio, 1.4), as did those who reported their pregnancy was unwanted (1.7) or mistimed (1.4). Among immigrant Latinas, those who reported being unsure about their pregnancy were at higher risk of preterm birth than were those who reported an intended pregnancy (1.6). After adjustment for socioeconomic factors, the association remained significant for immigrant Latinas who were unsure about their pregnancy (1.5), but none of the associations remained significant for whites.

CONCLUSIONS:  Women’s interpretations of questions about pregnancy intention and their social experiences regarding pregnancy intention may vary by race or ethnicity. Studies on the association between pregnancy intention and preterm birth may need to be group‐specific.
Pathways from Family Religiosity to Adolescent Sexual Activity and Contraceptive Use
Authors: Manlove Jennifer , Logan Cassandra , Moore Kristin A. , Ikramullah Erum
Published: June 2008

CONTEXT:  Few studies with nationally representative longitudinal data have examined whether and how family religiosity is associated with adolescent sexual and contraceptive behavior.

METHODS:  Data from the 1997 National Longitudinal Survey of Youth were used to examine associations between a multidimensional measure of family religiosity assessed during early adolescence and reproductive health outcomes (sexual activity, number of partners and consistent contraceptive use) at age 17. Pathways through which family religiosity is associated with these outcomes were identified using structural equation models.

RESULTS:  Family religiosity was negatively associated with adolescent sexual activity, both directly (beta, −0.14) and indirectly (−0.02). The indirect association was mediated by family cohesion (as reflected in parental monitoring among the entire sample and among males, and in parent‐teenager relationship quality and family routine activities among females) and negative peer behaviors. Greater family religiosity was indirectly associated with having fewer sexual partners (−0.03) and with using contraceptives consistently (0.02); these relationships were mediated through later age at first sex, more positive peer environments and higher levels of parental monitoring and awareness. However, among sexually active males (but not females), family religiosity was directly and negatively associated with contraceptive consistency (−0.11).

CONCLUSION:  Cohesive family environments and positive peer networks contribute to reduced levels of risky sexual behavior among adolescents from religious families. Parents who monitor their children’s activities and peer environments, engage their families in regular activities and foster strong parent‐child relationships can help reduce risky sexual behavior, regardless of family religiosity. Parental involvement in prevention programs may help reduce rates of teenage pregnancy and STDs.
Expression of the GLP-1 Receptor in Mouse, Rat, and Human Pancreas
Authors: Tornehave Ditte , Kristensen Peter , Rømer John , Knudsen Lotte Bjerre , Heller R. Scott
Published: June 2008

We studied the intra-islet localization of the glucagon-like peptide 1 receptor (GLP-1R) by colocalization studies of the GLP-1R mRNA and protein with islet cell hormones in mice, rats, and humans. In contrast to previous reports, we show that the GLP-1R is selectively located on the β cells. The localization of GLP-1R in islets and ducts was studied using ISH and double and triple fluorescence microscopy. In normal pancreatic tissue from mice and rats, GLP-1R mRNA was only detectable in the β cells. Double and triple immunofluorescence using two different GLP-1R antisera and combinations of insulin, glucagon, pancreatic polypeptide, and somatostatin showed that GLP-1R protein is almost exclusively colocalized with insulin. The same pattern was observed in human pancreas, but the GLP-1R expression was more heterogeneous, with populations of insulin immunoreactive cells with high and low expression. This is the first time that the GLP-1R has been localized in human islets. Furthermore, GLP-1R immunoreactivity was found in the pancreatic ducts in mouse, rat, and human pancreas. As an important confirmation of the specificity of our methods, we found no signals for GLP-1R mRNA or protein in pancreatic tissue from gene-targeted GLP-1R—deficient mice. In conclusion, our data suggest that the GLP-1 receptor is restricted to the pancreatic β cells and the lack of receptor immunoreactivity on δ cells cannot be explained suitably to correspond with published in vivo and in vitro data. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials.
Pax7 Shows Higher Satellite Cell Frequencies and Concentrations within Intrafusal Fibers of Muscle Spindles
Authors: Kirkpatrick Lisa J. , Allouh Mohammed Z. , Nightingale Chantale N. , Devon Heidi G. , Yablonka-Reuveni Zipora , Rosser Benjamin W.C.
Published: June 2008

Intrafusal fibers within muscle spindles make up a small subpopulation of muscle fibers. These proprioceptive fibers differ from most extrafusal fibers because, even in maturity, their diameters remain small, and they retain expression of developmental myosins. Although both extrafusal and intrafusal fibers contain satellite cells (SCs), comparatively little is known about intrafusal SCs. Analyzing chicken fast-phasic posterior (PLD) and slow-tonic anterior (ALD) latissimus dorsi muscles, we show that SCs of both intrafusal and extrafusal fibers express Pax7. We further test the hypotheses that intrafusal fibers display parameters reflective of extrafusal immaturity. These hypotheses are that intrafusal fibers contain (a) higher SC frequencies (number of SC nuclei/all nuclei within basal lamina) and concentrations (closer together) and (b) smaller myonuclear domains than do adjacent extrafusal fibers. IHC techniques were applied to PLD and ALD muscles excised at 30 and 138 days posthatch. The hypotheses were validated, suggesting that intrafusal fibers have greater capacities for growth, regeneration, and repair than do adjacent extrafusal fibers. During maturation, extrafusal and intrafusal fibers show similar trends of decreasing SC frequencies and concentrations and increases in myonuclear domains. Thus, extrafusal and intrafusal fibers alike should exhibit reduced capacities for growth, regeneration, and repair during maturation.
Music transmission in an Auckland Tongan community youth band
Authors: Hebert David G
Published: June 2008

This article reports on findings from a 2006 ethnographic study of a Tongan community youth band in Auckland, New Zealand. To begin, the study's rationale is discussed in relation to previous research, followed by a description of the band's repertoire and rehearsal strategies, instrumentation and uniforms, notational practices and institutional context. The youth band's role in the Tongan community of Auckland is then considered in relation to previous descriptions of community music. The band's significance in terms of musical identity and its socio-economic context are also examined, followed by a discussion of this study's implications for community music workers in other settings. The findings suggest that community ensembles rooted in musical hybridity may generate innovative models of music learning and play a unique role in cultural preservation.
Closing the gap: does music-making have to stop upon graduation?
Authors: Mantie Roger , Tucker Lynn
Published: June 2008

This paper discusses research examining the perceptions of people who continue to be musically active beyond their formal schooling years. These are individuals who are neither professional musicians nor university music majors; all pursue music avocationally. Through interviews and
focus groups with members of three community ensembles (directed by the researchers), we sought to better understand how these avocational musicians viewed their own reasons for participating in music-making, and what, if any, connection they perceived between their school music experience
and their present experience.
Lifelong learners in music; research into musicians' biographical learning
Authors: Smilde Rineke
Published: June 2008

This article considers four learning biographies from professional musicians. Each musician holds a different portfolio career and is from a different age category. The key questions I ask are: How does one learn as a musician?, What knowledge, attitudes, values and artistic skills
are necessary to function effectively and creatively as a contemporary musician?, and What is the necessary framework of lifelong learning in music education? My findings are analysed in the light of lifelong learning for musicians with a focus toward teaching and learning.