bluuebunny/crossref_metadata_2025 huggingface dataset:
Intrinsic Activity Rhythms in Macaca mulatta: Their Entrainment to Light and Melatonin
Authors: Masuda Ken , Zhdanova Irina V.
Published: September 2010

Mounting evidence that circadian abnormalities are a risk factor for cancer and for cardiovascular, psychiatric, and other disorders calls for in-depth investigation of intrinsic clock-dependent processes in diurnal animal models phylogenetically close to humans. Rhesus monkey ( Macaca mulatta) is the most extensively studied diurnal nonhuman primate. Similar to humans, it features consolidated nighttime sleep and advanced cardiovascular, neuroendocrine, and cognitive responses. However, the intrinsic circadian rhythmicity in this species remains to be fully characterized. Here it is demonstrated that under constant dim light (~10 lx) conditions, young adult rhesus monkeys maintain robust intrinsic circadian rhythms of activity, with periods ranging from 23.4 to 25.1 h. Constant environmental light of moderate intensity (~100 lx) slows down the circadian clock in rhesus monkeys. The exposure to light or melatonin shifts the phase of intrinsic circadian rhythms, with the direction and magnitude of the shift dependent on the circadian phase at which a stimulus was administered. The length of the intrinsic period largely defines an individual’s chronotype (morningness or eveningness) and affects the stability of intrinsic rhythms and the phase angle of entrainment to melatonin and light. This first detailed characterization of intrinsic circadian rhythms of activity and their responses to light and melatonin in rhesus monkeys shows principal similarities to those in humans. These findings should provide new opportunities for translational research on the effects of diverse agents, environmental conditions, aging, and disease on the circadian clock and its outputs.
The Social Clock of the Honeybee
Authors: Bloch Guy
Published: September 2010

The honeybee has long been an important model for studying the interplay between the circadian clock and complex behaviors. This article reviews studies further implicating the circadian clock in complex social behaviors in bees. The article starts by introducing honeybee social behavior and sociality and then briefly summarizes current findings on the molecular biology and neuroanatomy of the circadian system of honeybees that point to molecular similarities to the mammalian clockwork rather than to that of Drosophila. Foraging is a social behavior in honeybees that relies on the circadian clock for timing visits to flowers, time-compensated sun-compass navigation, and dance communication used by foragers to recruit nestmates to rewarding flower patches. The circadian clock is also important for the social organization of honeybee societies. Social factors influence the ontogeny of circadian rhythms and are important for social synchronization of worker activities. Both queen and worker bees switch between activities with and without circadian rhythms. In workers this remarkable plasticity is associated with the division of labor; nurse bees care for the brood around the clock with similar levels of clock gene expression throughout the day, whereas foragers have strong behavioral circadian rhythms with oscillating brain clock gene levels. This plasticity in circadian rhythms is regulated by direct contact with the brood and is context-specific in that nurse bees that are removed from the hive exhibit activity with strong behavioral and molecular rhythms. These studies on the sociochronobiology of honeybees and comparative studies with other social insects suggest that the evolution of sociality has influenced the characteristics of the circadian system in honeybees.
The Massachusetts Baby-Friendly Collaborative: Lessons Learned From an Innovation to Foster Implementation of Best Practices
Authors: Bartick Melissa , Edwards Roger A. , Walker Marsha , Jenkins Lucia
Published: September 2010

Collaborative efforts among hospitals can facilitate the exchange of ideas, provide a forum for discussing the development of new policies or practices or changes to existing policies and practices, and increase the implementation of best practices. In November 2008, the Massachusetts Breastfeeding Coalition formed a collaborative of maternity facilities wishing to pursue Baby-Friendly designation. Members provided insights from experiences and shared models and examples from outside. We describe highlights from the first 15 months of the Collaborative and present 4 recommendations for overcoming barriers: (1) manage expectations of patients, family/friends, and staff; (2) restrict access to materials that can undermine breastfeeding; (3) adopt the appropriate perspectives to creatively implement change; and (4) bundle, reframe, and harness larger forces. The strategies can be applied across diverse hospital settings.
Mitigation of arsenic-mediated renal oxidative stress in rat by Pleurotus florida lectin
Authors: Asit Kumar Bera  , Rana Tanmoy , Das Subhashree , Bhattacharya Debasis , Pan Diganta , Bandyopadhyay Subhasish , Subrata Kumar Das
Published: September 2010

Oyster mushroom, Pleurotus florida is regarded as one of the popular food with biopharmaceutical properties. Here, the study aimed to investigate the antioxidative effects of mushroom (Pleurotus florida) lectin against arsenic-induced nephrotoxicity in rats. Animals were divided into four groups; Group 1 was control. Groups 2, 3 and 4 were exposed to arsenic (20 parts per million [ppm] in drinking water), arsenic plus oral supplementation of ascorbic acid (25 mg/kg body weight) and arsenic plus oral supplementation of mushroom lectin (150 mg/kg body weight) respectively. Both ascorbic acid and mushroom lectin prevented the arsenic-mediated growth retardation and normalized the elevated kidney weight. Disrupted activities of superoxide dismutase (SOD) and catalase (CAT) and enhanced lipid peroxidation (LPO), protein carbonyl (PC) and nitric oxides (NO) production in kidney caused by arsenic could also be maintained towards normalcy by supplementation of mushroom lectin and ascorbic acid. These antioxidative effects were exhibited in a time-dependant manner. Further, arsenic-mediated down-regulation of messenger RNA (mRNA) expression of superoxide dismutase 2 (SOD2) gene was obstructed by these agents. Thus it was found that mushroom lectin reversed the effect of arsenic-mediated oxidative stress in a time-dependent manner.
“My Head is Like a Bag Full of Rubbish”: Concepts of Illness and Treatment Expectations in Traumatized Migrants
Authors: Maier Thomas , Straub Mirjam
Published: September 2010

Traumatized migrants increasingly burden health care services in industrialized countries. These patients usually suffer from a broad range of physical and psychological symptoms, with assessment and treatment being complicated and often unsatisfactory. A large potential for misunderstandings between clinicians and traumatized individuals is not only because of a language barrier; very little is known about the concepts of illness and treatment expectations in these patients. We performed semistructured interviews with 13 traumatized migrants, focusing on their concepts of illness and expectations concerning medical treatment. An analysis of the recorded and transcribed statements revealed that most participants had no clear or defined expectations concerning appropriate treatment. The patients knew very little about psychotherapy, and usually had negative prejudices. Nevertheless, participants firmly believed that medical treatment would help them and considered their current suffering to be the result of a multitude of stressors impacting their lives over long periods of time. The patients did not identify single traumatic events as the causes of their complaints. The prevailing concepts of illness among traumatized migrants can be characterized as multifactorial and psychosocial.
Review Article: Eplerenone: An Underused Medication?
Authors: Abuannadi Mohammad , O'Keefe James H.
Published: September 2010

In this article, we review the evidence supporting the use of eplerenone for improving cardiovascular prognosis. Activation of the renin-angiotensin-aldosterone system plays a major role in the pathogenesis of heart disease, and blockage of this system has been shown to improve prognosis in several cardiovascular conditions. The 2 marketed aldosterone antagonists, spironolactone and eplerenone, improve prognosis in patients with left ventricular (LV) dysfunction and are effective antihypertensive medications. In addition, a potential role for aldosterone antagonists in the treatment of patients with heart failure and preserved LV function has been suggested and is currently being evaluated in clinical trials. In patients with myocardial infarction having LV dysfunction and evidence of heart failure, eplerenone improves cardiovascular outcomes and attenuates myocardial remodeling. In addition, eplerenone is effective for the treatment of hypertension, where it regresses both LV hypertrophy and proteinuria (2 powerful markers of increased cardiovascular risk). In contrast to spironolactone, eplerenone essentially lacks the sexual side effects that sometimes limit the use of spironolactone. Hyperkalemia is the main potential side effect of eplerenone, especially when used in combination with other medications that can cause hyperkalemia. Adequate patient selection and monitoring are therefore of utmost importance when using this medication. In conclusion, eplerenone is a medication that offers the cardiovascular therapeutic and prognostic benefits of aldosterone antagonism but with fewer side effects compared to spironolactone.
Effects of High- and Low-Sodium Diets on Ambulatory Blood Pressure in Patients With Hypertension Receiving Aliskiren
Authors: Weir Matthew R. , Yadao Anthony M. , Purkayastha Das , Charney Alan N.
Published: September 2010

Dietary sodium reduction and, as necessary, pharmacologic treatment are recommended for hypertension management. This prospective, randomized, open-label, blinded-end point, multicenter, crossover study investigated the effect of dietary sodium intake on mean ambulatory systolic blood pressure (maSBP) in patients with hypertension receiving aliskiren 300 mg once daily. Following a 2- to 4-week washout period, patients were randomized to a high- (≥200 mmol/d) or low- (≤100 mmol/d) sodium diet and were started on aliskiren, 300 mg/d. After 4 weeks, patients were crossed over to the alternate diet for an additional 4 weeks. The primary efficacy variable was change in maSBP between diets. During treatment with aliskiren, maSBP was significantly lower with the low-sodium diet compared with the high-sodium diet (least squares mean difference, 9.4 mm Hg; 95% CI, 7.5-11.4; P < .0001). The percentage of patients achieving a maSBP response to aliskiren (<130 mm Hg or a ≥20-mm Hg reduction from baseline) was greater with the low- (76.5%) versus the high-sodium diet (42.6%; P < .0001). Overall, 40.9% patients had ≥1 adverse event and the rates were similar between groups. In this study, aliskiren was well tolerated and a low-sodium diet accentuated its antihypertensive effect.
The autism diagnostic experiences of French parents
Authors: Chamak Brigitte , Bonniau Béatrice , Oudaya Lila , Ehrenberg Alain
Published: September 2010

This survey focused on French parents’ views of the diagnostic process relating to their child with autism. Data were collected on the age at diagnosis, the time taken to obtain a diagnosis and the difficulties encountered. Questionnaires filled in by the parents ( n = 248) and in-depth interviews ( n = 43) were analyzed in order to obtain quantitative and qualitative results. The ages of the children ranged from 4 to 45. This approach enabled us to compare the practices of professionals now and in the past. The mean age of diagnosis was 10 ± 8 years from 1960 to 1990, 5 ± 3 years from 1990 to 2005 (3 ± 1 from 2003 to 2005). The results showed that the mean delays between first consultation and diagnosis were reduced. Regarding the way the diagnosis was announced, 63% of the parents of children with autism and 93% of the parents of adults with autism were dissatisfied. We discuss the parents’ reactions and the changes in the diagnostic process.
Evaluating lexical cohesion in telephone conversations
Authors: Gómez González María de los Ángeles
Published: September 2010

Ever since the publication of Halliday and Hasan (1976), cohesion analysis has received much attention in several branches of linguistics. Lexical cohesion in particular has been shown to contribute to the coherence of discourse in a number of ways (Ferstl and von Cramon, 2001; Hellman, 1995; Hoey, 1991b; Sanders and Pander Maat, 2006), and specific patterns of lexical cohesion have emerged as relevant for the description of different registers and genres (Louwerse et al ., 2004; Taboada, 2004; Tanskanen, 2006; Thompson, 1994). In the present article I challenge existing models of lexical cohesion and offer a revised one which affords particular attention to what I call ‘associative cohesion’. I go on to test the adequacy of the proposed framework in a corpus of 15 telephone conversations (20,043 words) extracted from the International Corpus of English-Great Britain. Using both quantitative and qualitative methods the article analyses 3480 ties and demonstrates that telephone conversations are lexically cohesive mostly due to repetitions (52.6%), associative cohesion (24%) and inclusive relations (10.5%), which overwhelmingly occur across the turns (87.6%) and over remote-mediated spans (71.9%). In addition, lexical patterns are shown to collaborate in topic management processes and to be sensitive to genre-specific factors, thereby demonstrating the descriptive potential and applicability of the framework.
Comparing rhetorical structures in different languages: The influence of translation strategies
Authors: da Cunha Iria , Iruskieta Mikel
Published: September 2010

The study we report in this article addresses the results of comparing the rhetorical trees from two different languages carried out by two annotators starting from the Rhetorical Structure Theory (RST). Furthermore, we investigate the methodology for a suitable evaluation, both quantitative and qualitative, of these trees. Our corpus contains abstracts of medical research articles written both in Spanish and Basque, and extracted from Gaceta Médica de Bilbao (‘Medical Journal of Bilbao’). The results demonstrate that almost half of the annotator disagreement is due to the use of translation strategies that notably affect rhetorical structures.
Lessons learned from implementing a psycho-educational intervention for African American dementia caregivers
Authors: Morano Carmen L. , King M. Denise
Published: September 2010

Despite the fact that African American, Latino, and Asian Americans represent the fastest growing segments of the population in the United States over the age of 65, they remain understudied in intervention research. This article describes the process of developing and implementing a psycho-educational intervention for African American caregivers which was informed by a review of the caregiving and intervention literature, and stress, appraisal, and coping theory. The intervention included 12 modules that lasted approximately 90 minutes each. Participants were recruited from the local Alzheimer’s Association, local area churches and social clubs, and primary care physicians. The article discusses issues that came up during the pilot test and highlights the importance of participant input, in the design, implementation, and evaluation of intervention programmes.
News Values and Resource Allocation as the Determinants of Election Coverage: Analysis of a Hong Kong Election
Authors: Lee Francis L. F.
Published: September 2010

While past research has shown that the news media can mobilize citizens for voting participation, little has been done on the determinants of the level of attention paid by the media to elections. Drawing on theories of news production, this study postulates that considerations of news values and the dynamics of resource allocation would combine to shape the quantity of election news. The empirical analysis focuses on a local election in Hong Kong, in which the news media paid different levels of attention to the contests in different constituencies. It shows that characteristics of the electoral contests relevant to the news values of prominence and conflict can explain how much coverage a constituency received, and the influences of news values factors can be interactive as well as additive. At the same time, the influence of resource allocation is illustrated by a relatively stronger media focus on the contests in urban center areas as well as the support for a resource spillover hypothesis. The general implications of the findings are discussed.
Who Tube? How YouTube’s News and Politics Space Is Going Mainstream
Authors: May Albert L.
Published: September 2010

In its drive to make a profit and avoid copyright challenges, YouTube, the largest online video platform, is becoming a more hospitable place for news organizations that are targeting the medium’s young audience with an attractive fare of raw video and light content. But this changing YouTube environment appears to be less welcoming to the politically motivated online operations that found a voice there in the 2008 presidential election. This article explores the developments and tracks the audience changes to the largest YouTube news and politics sites from just before the election through early 2010. Corporate media were found to be more successful in building large sustained audiences, while online-only operations with a political bent have lost audience share since the election and appear to be confined to a niche.
Health Care Reform and Asthma Care Disparities in Minority Populations
Authors: Cambridge Lisa
Published: September 2010

Asthma affects almost 23 million people in the United States and ranks as a leading cause of health disparities among minority and low socioeconomic groups, with the largest impact on minority children. This article will examine how fragmented health care in the United States contributes to asthma care disparities in minority populations and will discuss how health care reform may help overcome these disparities. Causes of asthma care disparities are marked by the socioeconomic relationship of the disease including, but not limited to, underprescribing and underuse of preventative medication, low primary care follow-up after acute exacerbations, and the limited availability of pharmacy resources in urban communities. Furthermore, the importance of health care coverage cannot be overemphasized. With the recent passage of the Patient Protection and Affordable Care Act, health care reform offers significant opportunities to reduce or eliminate disparities in minorities. For health care reform to succeed, it must address the specific needs of minorities and support the development of a wide range of community-based interventions and programs with an emphasis on prevention, primary care, chronic care management, and care coordination. In addition, policies that promote cultural competence and improve access and continuity of care are needed, as they serve as predictors of higher-quality asthma care in minorities. Current asthma initiatives targeting minorities need better coordination and increased funding. Solutions to improve adherence to evidence-based clinical practice guidelines may be a potentially powerful tool for reducing or eliminating disparities in asthma care. For asthma care interventions and policies to be effective in targeting asthma care disparities, they must be implemented in a variety of health care settings.
A CpG-loaded tumor cell vaccine induces antitumor CD4+ T cells that are effective in adoptive therapy for large and established tumors
Authors: Goldstein Matthew J. , Varghese Bindu , Brody Joshua D. , Rajapaksa Ranjani , Kohrt Holbrook , Czerwinski Debra K. , Levy Shoshana , Levy Ronald
Published: September 2010

We designed a whole tumor cell vaccine by “loading” lymphoma tumor cells with CG-enriched oligodeoxynucleotide (CpG), a ligand for the Toll-like receptor 9 (TLR9). CpG-loaded tumor cells were phagocytosed, delivering both tumor antigen(s) and the immunostimulatory CpG molecule to antigen-presenting cells (APCs). These APCs then expressed increased levels of costimulatory molecules and induced T-cell immunity. TLR9 was required in the APCs but not in the CpG-loaded tumor cell. We demonstrate that T cells induced by this vaccine are effective in adoptive cellular therapy for lymphoma. T cells from vaccinated mice transferred into irradiated, syngeneic recipients protected against subsequent lymphoma challenge and, remarkably, led to regression of large and established tumors. This therapeutic effect could be transferred by CD4+ but not by CD8+ T cells. A CpG-loaded whole-cell vaccination is practical and has strong potential for translation to the clinical setting. It is currently being tested in a clinical trial of adoptive immunotherapy for mantle-cell lymphoma.
Zebrafish fin immune responses during high mortality infections with viral haemorrhagic septicemia rhabdovirus. A proteomic and transcriptomic approach
Authors: Encinas Paloma , Rodriguez-Milla Miguel A , Novoa Beatriz , Estepa Amparo , Figueras Antonio , Coll Julio
Published: September 2010

Background

Despite rhabdoviral infections being one of the best known fish diseases, the gene expression changes induced at the surface tissues after the natural route of infection (infection-by-immersion) have not been described yet. This work describes the differential infected versus non-infected expression of proteins and immune-related transcripts in fins and organs of zebrafish Danio rerio shortly after infection-by-immersion with viral haemorrhagic septicemia virus (VHSV).

Results

Two-dimensional differential gel electrophoresis detected variations on the protein levels of the enzymes of the glycolytic pathway and cytoskeleton components but it detected very few immune-related proteins. Differential expression of immune-related gene transcripts estimated by quantitative polymerase chain reaction arrays and hybridization to oligo microarrays showed that while more transcripts increased in fins than in organs (spleen, head kidney and liver), more transcripts decreased in organs than in fins. Increased differential transcript levels in fins detected by both arrays corresponded to previously described infection-related genes such as complement components (c3b, c8 and c9) or class I histocompatibility antigens (mhc1) and to newly described genes such as secreted immunoglobulin domain (sid4), macrophage stimulating factor (mst1) and a cluster differentiation antigen (cd36).

Conclusions

The genes described would contribute to the knowledge of the earliest molecular events occurring in the fish surfaces at the beginning of natural rhabdoviral infections and/or might be new candidates to be tested as adjuvants for fish vaccines.
Burst expansion, distribution and diversification of MITEs in the silkworm genome
Authors: Han Min-Jin , Shen Yi-Hong , Gao Ying-Hui , Chen Li-Yong , Xiang Zhong-Huai , Zhang Ze
Published: September 2010

Background

Miniature inverted-repeat transposable elements (MITEs) are widespread in plants and animals. Although silkworm (Bombyx mori) has a large amount of and a variety of transposable elements, the genome-wide information of the silkworm MITEs is unknown.

Results

We used structure-based and homology approaches to search for MITEs in the silkworm genome. We identified 17 MITE families with a total of 5785 members, accounting for ~0.4% of the genome. 7 of 17 MITE families are completely novel based on the nucleotide composition of target site duplication (TSD) and/or terminal inverted repeats (TIR). Silkworm MITEs were widely and nonrandom distributed in the genome. One family named BmMITE-2 might experience a recent burst expansion. Network and diversity analyses for each family revealed different diversification patterns of the silkworm MITEs, reflecting the signatures of genome-shocks that silkworm experienced. Most silkworm MITEs preferentially inserted into or near genes and BmMITE-11 that encodes a germline-restricted small RNA might silence its the closest genes in silkworm ovary through a small RNA pathway.

Conclusions

Silkworm harbors 17 MITE families. The silkworm MITEs preferred to reside in or near genes and one MITE might be involved in gene silence. Our results emphasize the exceptional role of MITEs in transcriptional regulation of genes and have general implications to understand interaction between MITEs and their host genome.
Overproduced Brucella abortus PdhS-mCherry forms soluble aggregates in Escherichia coli, partially associating with mobile foci of IbpA-YFP
Authors: Van der Henst Charles , Charlier Caroline , Deghelt Michaël , Wouters Johan , Matroule Jean-Yves , Letesson Jean-Jacques , De Bolle Xavier
Published: September 2010

Background

When heterologous recombinant proteins are produced in Escherichia coli, they often precipitate to form insoluble aggregates of unfolded polypeptides called inclusion bodies. These structures are associated with chaperones like IbpA. However, there are reported cases of "non-classical" inclusion bodies in which proteins are soluble, folded and active.

Results

We report that the Brucella abortus PdhS histidine kinase fused to the mCherry fluorescent protein forms intermediate aggregates resembling "non-classical" inclusion bodies when overproduced in E. coli, before forming "classical" inclusion bodies. The intermediate aggregates of PdhS-mCherry are characterized by the solubility of PdhS-mCherry, its ability to specifically recruit known partners fused to YFP, suggesting that PdhS is folded in these conditions, and the quick elimination (in less than 10 min) of these structures when bacterial cells are placed on fresh rich medium. Moreover, soluble PdhS-mCherry foci do not systematically colocalize with IpbA-YFP, a marker of inclusion bodies. Instead, time-lapse experiments show that IbpA-YFP exhibits rapid pole-to-pole shuttling, until it partially colocalizes with PdhS-mCherry aggregates.

Conclusion

The data reported here suggest that, in E. coli, recombinant proteins like PdhS-mCherry may transit through a soluble and folded state, resembling previously reported "non-classical" inclusion bodies, before forming "classical" inclusion bodies. The dynamic localization of IbpA-YFP foci suggests that the IbpA chaperone could scan the E. coli cell to find its substrates.
Detection of sexually transmitted infection and human papillomavirus in negative cytology by multiplex-PCR
Authors: Shim Hyo-Sub , Noh Songmi , Park Ae-Ran , Lee Young-Nam , Kim Jong-Kee , Chung Hyun-Jae , Kang Keum-Soon , Cho Nam Hoon
Published: September 2010

Background

The aim of this study was to determine the prevalence of human papillomavirus (HPV) and 15 species that cause sexually transmitted infections (STIs) in negative cytology. In addition, we compared the diagnostic performance of multiplex polymerase chain reaction (PCR) with widely available techniques used to detect HPV.

Methods

We recruited 235 women of reproductive age who had negative cytology findings in a liquid-based cervical smear. STIs were identified by multiplex PCR, and HPV genotypes by multiplex PCR, hybrid capture 2, and DNA microaray; discordant results were analyzed by direct sequencing.

Results

Approximately 96.6% of patients with negative cytology results were positive for pathogens that cause STIs. The pathogens most frequently detected were Gardnerella vaginalis, Ureaplasma urealyticum. The incidence of HPV in negative cytology was 23.3%. Low-risk HPV infection was significantly correlated with Chalmaydia trachomatis, and high-risk HPV infection was significantly correlated with Group β streptococcus. The analytical sensitivities of the multiplex PCR and DNA microarray were higher than 80%, and the analytical specificity was nearly 100% for all tests.

Conclusions

Multiplex PCR yielded results that most of patients with negative cytology were positive for pathogens that cause STIs, and were more similar to that of DNA microarray, than that of hybrid capture 2 in terms of analytical sensitivity and prediction value of HPV infection.
Transcriptional profiling of ErbB signalling in mammary luminal epithelial cells - interplay of ErbB and IGF1 signalling through IGFBP3 regulation
Authors: Worthington Jenny , Bertani Mariana , Chan Hong-Lin , Gerrits Bertran , Timms John F
Published: September 2010

Background

Members of the ErbB family of growth factor receptors are intricately linked with epithelial cell biology, development and tumourigenesis; however, the mechanisms involved in their downstream signalling are poorly understood. Indeed, it is unclear how signal specificity is achieved and the relative contribution each receptor has to specific gene expression.

Methods

Gene expression profiling of a human mammary luminal epithelial cell model of ErbB2-overexpression was carried out using cDNA microarrays with a common RNA reference approach to examine long-term overlapping and differential responses to EGF and heregulin beta1 treatment in the context of ErbB2 overexpression. Altered gene expression was validated using quantitative real time PCR and/or immunoblotting. One gene of interest was targeted for further characterisation, where the effects of siRNA-mediated silencing on IGF1-dependent signalling and cellular phenotype were examined and compared to the effects of loss of ErbB2 expression.

Results

775 genes were differentially expressed and clustered in terms of their growth factor responsiveness. As well as identifying uncharacterized genes as novel targets of ErbB2-dependent signalling, ErbB2 overexpression augmented the induction of multiple genes involved in proliferation (e.g. MYC, MAP2K1, MAP2K3), autocrine growth factor signalling (VEGF, PDGF) and adhesion/cytoskeletal regulation (ZYX, THBS1, VCL, CNN3, ITGA2, ITGA3, NEDD9, TAGLN), linking them to the hyper-poliferative and altered adhesive phenotype of the ErbB2-overexpressing cells. We also report ErbB2-dependent down-regulation of multiple interferon-stimulated genes that may permit ErbB2-overexpressing cells to resist the anti-proliferative action of interferons. Finally, IGFBP3 was unique in its pattern of regulation and we further investigated a possible role for IGFBP3 down-regulation in ErbB2-dependent transformation through suppressed IGF1 signalling. We show that IGF1-dependent signalling and proliferation were enhanced in ErbB2-overexpressing cells, whilst loss of ErbB2 expression by siRNA silencing reduced IGF1 signalling. Furthermore, IGFBP3 knockdown resulted in basal ERK and Akt activation in luminal epithelial cells and increased invasiveness and anchorage-independent colony formation in SKBR3 cells.

Conclusions

These data show IGFBP3 as a negative regulator of transformation and that its down-regulation enhances IGF1-dependent signalling. They also show that ErbB2 can up-regulate IGF1-dependent signalling, possibly via the regulated expression of IGFBP3.
Hyaluronan synthases (HAS1-3) and hyaluronidases (HYAL1-2) in the accumulation of hyaluronan in endometrioid endometrial carcinoma
Authors: Nykopp Timo K , Rilla Kirsi , Tammi Markku I , Tammi Raija H , Sironen Reijo , Hämäläinen Kirsi , Kosma Veli-Matti , Heinonen Seppo , Anttila Maarit
Published: September 2010

Background

Hyaluronan accumulation correlates with the degree of malignancy in many solid tumor types, including malignant endometrial carcinomas. To elucidate the mechanism of hyaluronan accumulation, we examined the expression levels of the hyaluronan synthases (HAS1, HAS2 and HAS3) and hyaluronidases (HYAL1 and HYAL2), and correlated them with hyaluronan content and HAS1-3 immunoreactivity.

Methods

A total of 35 endometrial tissue biopsies from 35 patients, including proliferative and secretory endometrium (n = 10), post-menopausal proliferative endometrium (n = 5), complex atypical hyperplasia (n = 4), grade 1 (n = 8) and grade 2 + 3 (n = 8) endometrioid adenocarcinomas were divided for gene expression by real-time RT-PCR, and paraffin embedded blocks for hyaluronan and HAS1-3 cytochemistry.

Results

The mRNA levels of HAS1-3 were not consistently changed, while the immunoreactivity of all HAS proteins was increased in the cancer epithelium. Interestingly, HAS3 mRNA, but not HAS3 immunoreactivity, was increased in post-menopausal endometrium compared to normal endometrium (p = 0.003). The median of HYAL1 mRNA was 10-fold and 15-fold lower in both grade 1 and grade 2+3 endometrioid endometrial cancers, as compared to normal endometrium (p = 0.004-0.006), and post-menopausal endometrium (p = 0.002), respectively. HYAL2 mRNA was also reduced in cancer (p = 0.02) and correlated with HYAL1 (r = 0.8, p = 0.0001). There was an inverse correlation between HYAL1 mRNA and the epithelial hyaluronan staining intensity (r = -0.6; P = 0.001).

Conclusion

The results indicated that HYAL1 and HYAL2 were coexpressed and significantly downregulated in endometrioid endometrial cancer and correlated with the accumulation of hyaluronan. While immunoreactivity for HASs increased in the cancer cells, tumor mRNA levels for HASs were not changed, suggesting that reduced turnover of HAS protein may also have contributed to the accumulation of hyaluronan.
Short-hairpin RNA-induced suppression of adenine nucleotide translocase-2 in breast cancer cells restores their susceptibility to TRAIL-induced apoptosis by activating JNK and modulating TRAIL receptor expression
Authors: Jang Ji-Young , Jeon Yoon-Kyung , Choi Yun , Kim Chul-Woo
Published: September 2010

Background

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL; apo2 ligand) induces apoptosis in cancer cells but has little effect on normal cells. However, many cancer cell types are resistant to TRAIL-induced apoptosis, limiting the clinical utility of TRAIL as an anti-cancer agent. We previously reported that the suppression of adenine nucleotide translocase-2 (ANT2) by short-hairpin RNA (shRNA) induces apoptosis of breast cancer cells, which frequently express high levels of ANT2. In the present study, we examined the effect of RNA shRNA-induced suppression of ANT2 on the resistance of breast cancer cells to TRAIL-induced apoptosis in vitro and in vivo.

Results

ANT2 shRNA treatment sensitized MCF7, T47 D, and BT474 cells to TRAIL-induced apoptosis by up-regulating the expression of TRAIL death receptors 4 and 5 (DR4 and DR5) and down-regulating the TRAIL decoy receptor 2 (DcR2). In MCF7 cells, ANT2 knockdown activated the stress kinase c-Jun N-terminal kinase (JNK), subsequently stabilizing and increasing the transcriptional activity of p53 by phosphorylating it at Thr81; it also enhanced the expression and activity of DNA methyltransferase 1 (DNMT1). ANT2 shRNA-induced overexpression of DR4/DR5 and TRAIL sensitization were blocked by a p53 inhibitor, suggesting that p53 activation plays an important role in the transcriptional up-regulation of DR4/DR5. However, ANT2 knockdown also up-regulated DR4/DR5 in the p53-mutant cell lines BT474 and T47 D. In MCF7 cells, ANT2 shRNA treatment led to DcR2 promoter methylation and concomitant down-regulation of DcR2 expression, consistent with the observed activation of DNMT1. Treatment of the cells with a demethylating agent or JNK inhibitor prevented the ANT2 shRNA-induced down-regulation of DcR2 and activation of both p53 and DNMT1. In in vivo experiments using nude mice, ANT2 shRNA caused TRAIL-resistant MCF7 xenografts to undergo TRAIL-induced cell death, up-regulated DR4/DR5, and down-regulated DcR2. Co-treatment with ANT2 shRNA and TRAIL efficiently suppressed tumor growth in these mice.

Conclusions

ANT2 suppression by shRNA might be exploited to overcome TRAIL-resistance in cancer.
Protease Activated Receptors 1 and 4 Sensitize TRPV1 in Nociceptive Neurones
Authors: Vellani Vittorio , Kinsey Anna M , Prandini Massimiliano , Hechtfischer Sabine C , Reeh Peter , Magherini Pier C , Giacomoni Chiara , McNaughton Peter A
Published: September 2010

Protease-activated receptors (PAR1–4) are activated by proteases released by cell damage or blood clotting, and are known to be involved in promoting pain and hyperalgesia. Previous studies have shown that PAR2 receptors enhance activation of TRPV1 but the role of other PARs is less clear. In this paper we investigate the expression and function of the PAR1, 3 and 4 thrombin-activated receptors in sensory neurones. Immunocytochemistry and in situ hybridization show that PAR1 and PAR4 are expressed in 10 – 15% of neurons, distributed across all size classes. Thrombin or a specific PAR1 or PAR4 activating peptide (PAR1/4-AP) caused functional effects characteristic of activation of the PLCp/PKC pathway: intracellular calcium release, sensitisation of TRPV1, and translocation of the epsilon isoform of PKC (PKCe) to the neuronal cell membrane. Sensitisation of TRPV1 was significantly reduced by PKC inhibitors. Neurons responding to thrombin or PAR1-AP were either small nociceptive neurones of the peptidergic subclass, or larger neurones which expressed markers for myelinated fibres. Sequential application of PAR1-AP and PAR4-AP showed that PAR4 is expressed in a subset of the PAR1-expressing neurons. Calcium responses to PAR2-AP were by contrast seen in a distinct population of small IB4+ nociceptive neurones. PAR3 appears to be non-functional in sensory neurones. In a skin-nerve preparation the release of the neuropeptide CGRP by heat was potentiated by PAR1-AP. Culture with nerve growth factor (NGF) increased the proportion of thrombin-responsive neurons in the IB4- population, while glial-derived neurotropic factor (GDNF) and neurturin upregulated the proportion of thrombin-responsive neurons in the IB4+ population. We conclude that PAR1 and PAR4 are functionally expressed in large myelinated fibre neurons, and are also expressed in small nociceptors of the peptidergic subclass, where they are able to potentiate TRPV1 activity.
On the traces of XPD: cell cycle matters - untangling the genotype-phenotype relationship of XPD mutations
Authors: Cameroni Elisabetta , Stettler Karin , Suter Beat
Published: September 2010

Mutations in the human gene coding for XPD lead to segmental progeria - the premature appearance of some of the phenotypes normally associated with aging - which may or may not be accompanied by increased cancer incidence. XPD is required for at least three different critical cellular functions: in addition to participating in the process of nucleotide excision repair (NER), which removes bulky DNA lesions, XPD also regulates transcription as part of the general transcription factor IIH (TFIIH) and controls cell cycle progression through its interaction with CAK, a pivotal activator of cyclin dependent kinases (CDKs). The study of inherited XPD disorders offers the opportunity to gain insights into the coordination of important cellular events and may shed light on the mechanisms that regulate the delicate equilibrium between cell proliferation and functional senescence, which is notably altered during physiological aging and in cancer.

The phenotypic manifestations in the different XPD disorders are the sum of disturbances in the vital processes carried out by TFIIH and CAK. In addition, further TFIIH- and CAK-independent cellular activities of XPD may also play a role. This, added to the complex feedback networks that are in place to guarantee the coordination between cell cycle, DNA repair and transcription, complicates the interpretation of clinical observations. While results obtained from patient cell isolates as well as from murine models have been elementary in revealing such complexity, the Drosophila embryo has proven useful to analyze the role of XPD as a cell cycle regulator independently from its other cellular functions. Together with data from the biochemical and structural analysis of XPD and of the TFIIH complex these results combine into a new picture of the XPD activities that provides ground for a better understanding of the patophysiology of XPD diseases and for future development of diagnostic and therapeutic tools.
First-time patellar dislocation with resultant habitual dislocation two years later, which was not demonstrated on plain X-rays halfway: a case report
Authors: Ohki Satoshi , Enomoto Hiroyuki , Nomura Eiki , Tanikawa Hidenori , Niki Yasuo , Matsumoto Hideo , Toyama Yoshiaki , Suda Yasunori
Published: September 2010

We present an instructive case of habitual left patellar dislocation in which the patella had appeared odd due to lateral tilt relative to contralateral side, but had been radiologically confirmed to be on the trochlea at 1 year prior to the referral. An 11-year-old girl presented to our hospital 2 years after the left patella had dislocated with a 'giving way' when cutting to the left. Our physical and radiological examinations confirmed that the left patella was laterally tilted in the patellar groove with the knee in extension but was dislocated in flexion beyond 45°. In spite of these findings, she had been untreated at the previous hospital since all plain X-rays, including a skyline patellar view, had failed to demonstrate the dislocation. Consequently, in addition to reconstruction of medial patellofemoral ligament, she had to undergo a lateral retinacular release, which might have been unnecessary if treated earlier. This case illustrates that first-time patellar dislocation can gradually lead to habitual dislocation subsequently, and that cautious physical examinations in regard to patella tracking are essential since radiological examinations do not always reveal the pathophysiology of patellar instability.
Extreme loss of immunoreactive p-Akt and p-Erk1/2 during routine fixation of primary breast cancer
Authors: Pinhel Isabel F , MacNeill Fiona A , Hills Margaret J , Salter Janine , Detre Simone , A'Hern Roger , Nerurkar Ashutosh , Osin Peter , Smith Ian E , Dowsett Mitch
Published: September 2010

Introduction

Very few studies have investigated whether the time elapsed between surgical resection and tissue fixation or the difference between core-cut and excision biopsies impact on immunohistochemically measured biomarkers, including phosphorylated proteins in primary breast cancer. The aim of this study was to characterise the differences in immunoreactivity of common biomarkers that may occur (1) as a result of tissue handling at surgery and (2) between core-cuts and resected tumours.

Methods

Core-cuts taken from surgical breast cancer specimens immediately after resection (sample A) and after routine X-ray of the excised tumour (sample B) were formalin-fixed and paraffin-embedded and compared with the routinely fixed resection specimen (sample C). The variation in immunohistochemical expression of Ki67, oestrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor 2 (HER2), p-Akt and p-Erk1/2 were investigated.

Results

Twenty-one tissue sets with adequate tumour were available. Median time between collection of core-cuts A and B was 30 minutes (range, 20 to 80 minutes). None of the markers showed significant differences between samples A and B. Similarly, Ki67, ER, PgR and HER2 did not differ significantly between core-cuts and main resection specimen, although there was a trend for lower resection values for ER (P = 0.06). However, p-Akt and p-Erk1/2 were markedly lower in resections than core-cuts (median, 27 versus 101 and 69 versus 193, respectively; both P < 0.0001 [two-sided]). This difference was significantly greater in mastectomy than in lumpectomy specimens for p-Erk1/2 (P = 0.01).

Conclusions

The delay in fixation in core-cuts taken after postoperative X-ray of resection specimens has no significant impact on expression of Ki67, ER, PgR, HER2, p-Akt or p-Erk1/2. However, extreme loss of phospho-staining can occur during routine fixation of resection specimens. These differences are likely attributable to suboptimal fixation and may have major repercussions for clinical research involving these markers.
Genome-Wide Associations and Functional Genomic Studies of Musculoskeletal Adverse Events in Women Receiving Aromatase Inhibitors
Authors: Ingle James N. , Schaid Daniel J. , Goss Paul E. , Liu Mohan , Mushiroda Taisei , Chapman Judy-Anne W. , Kubo Michiaki , Jenkins Gregory D. , Batzler Anthony , Shepherd Lois , Pater Joseph , Wang Liewei , Ellis Matthew J. , Stearns Vered , Rohrer Daniel C. , Goetz Matthew P. , Pritchard Kathleen I. , Flockhart David A. , Nakamura Yusuke , Weinshilboum Richard M.
Published: September 2010

Purpose

We performed a case-control genome-wide association study (GWAS) to identify single nucleotide polymorphisms (SNPs) associated with musculoskeletal adverse events (MS-AEs) in women treated with aromatase inhibitors (AIs) for early breast cancer.

Patients and Methods

A nested case-control design was used to select patients enrolled onto the MA.27 phase III trial comparing anastrozole with exemestane. Cases were matched to two controls and were defined as patients with grade 3 or 4 MS-AEs (according to the National Cancer Institute's Common Terminology Criteria for Adverse Events v3.0) or those who discontinued treatment for any grade of MS-AE within the first 2 years. Genotyping was performed with the Illumina Human610-Quad BeadChip.

Results

The GWAS included 293 cases and 585 controls. A total of 551,358 SNPs were analyzed, followed by imputation and fine mapping of a region of interest on chromosome 14. Four SNPs on chromosome 14 had the lowest P values (2.23E-06 to 6.67E-07). T-cell leukemia 1A (TCL1A) was the gene closest (926-7000 bp) to the four SNPs. Functional genomic studies revealed that one of these SNPs (rs11849538) created an estrogen response element and that TCL1A expression was estrogen dependent, was associated with the variant SNP genotypes in estradiol-treated lymphoblastoid cells transfected with estrogen receptor alpha and was directly related to interleukin 17 receptor A (IL17RA) expression.

Conclusion

This GWAS identified SNPs associated with MS-AEs in women treated with AIs and with a gene (TCL1A) which, in turn, was related to a cytokine (IL17). These findings provide a focus for further research to identify patients at risk for MS-AEs and to explore the mechanisms for these adverse events.
Value of Early Referral to Fertility Preservation in Young Women With Breast Cancer
Authors: Lee Sanghoon , Ozkavukcu Sinan , Heytens Elke , Moy Fred , Oktay Kutluk
Published: September 2010

Purpose

To determine whether early referral to reproductive specialists improves fertility preservation (FP) outcomes and reduces delay in adjuvant treatment in young women with breast cancer.

Patients and Methods

A secondary analysis of a prospective database of patients with breast cancer undergoing ovarian stimulation (OS) for FP by oocyte or embryo cryopreservation was performed.

Results

Of the 154 patients, 93 met the inclusion criteria (mean age, 35.2 ± 4.4 years). Thirty-five of the 93 patients were referred before breast surgery (PreS), and 58 patients were referred after surgery (PostS). The time periods from initial diagnosis (ID) to initiation of OS (42.6 ± 27.7 days for PreS v 71.9 ± 30.7 days for PostS; P < .001) and from ID to initiation of chemotherapy (83.9 ± 24.3 days for PreS v 107.8 ± 42.9 days for PostS; P = .045) were significantly shorter for the PreS group versus the PostS group. Nine (25.7%) of 35 patients in the PreS group versus one (1.7%) of 58 patients in the PostS group were able to undergo two FP cycles (P < .001), resulting in an increased yield of oocytes in the PreS group (18.2% [93 of 511 oocytes] v 0.6% [five of 800 oocytes], respectively; P < .001) and embryos (17.2% [40 of 233 embryos] v 0.6% [two of 357 embryos], respectively; P < .001). Patients who had an oocyte retrieval within 5 weeks of the surgery were able to complete a second cycle within 9 weeks of the surgery.

Conclusion

FP referral before breast surgery enables earlier initiation of cryopreservation cycles and chemotherapy and, when appropriate, multiple FP cycles. Women who can undergo multiple cycles may be at advantage for FP because of a larger number of oocytes or embryos cryopreserved. This is the first study demonstrating the benefit of early FP referral in patients with cancer.