bluuebunny/crossref_metadata_2025 huggingface dataset:
Best practice in the treatment of nonmuscle invasive bladder cancer
Authors: Anastasiadis Anastasios , de Reijke Theo M.
Published: December 2011

Bladder carcinoma is the most common malignancy of the urinary tract. Approximately 75–85% of patients with bladder cancer present with a disease that is confined to the mucosa (stage Ta, carcinoma in situ) or submucosa (stage T1). These categories are grouped as nonmuscle invasive bladder cancer (NMIBC). Although the management of NMIBC tumours has significantly improved during the past few years, it remains difficult to predict the heterogeneous outcome of such tumours, especially if high-grade NMIBC is present. Transurethral resection is the initial treatment of choice for NMIBC. However, the high rates of recurrence and significant risk of progression in higher-grade tumours mandate additional therapy with intravesical agents. We discuss the role of various intravesical agents currently in use, including the immunomodulating agent bacillus Calmette-Guérin (BCG) and chemotherapeutic agents. We also discuss the current guidelines and the role of these therapeutic agents in the context of higher-grade Ta and T1 tumours. Beyond the epidemiology, this article focuses on the risk factors, classification and diagnosis, the prediction of recurrence and progression in NMIBC, and the treatments advocated for this invasive disease.
Emerging novel therapies for advanced prostate cancer
Authors: Osanto Susanne , Van Poppel Hendrik
Published: December 2011

This review examines the development and efficacy of novel treatment options for advanced prostate cancer and discusses novel therapies that are on the horizon. Since the introduction of docetaxel as the standard treatment for patients with metastatic castration-resistant prostate cancer (CRPC), a number of different agents have been tested but failed to demonstrate improvement in overall survival (OS). Recently, three novel compounds have demonstrated OS benefit and one other showed reduction in skeletal-related events (SREs). Sipuleucel-T, a novel vaccine, was approved by the US regulatory authorities in April 2010 for patients with early advanced prostate cancer. A new taxane, cabazitaxel, and abiraterone acetate, an androgen biosynthesis inhibitor, have shown an OS benefit in advanced CRPC after docetaxel, leading to drug approval. A new bone-targeting agent, denosumab, a receptor activator of nuclear factor κB ligand (RANKL) antagonist, showed a modest reduction in SREs in comparison to zoledronic acid in patients with bone metastases. Other promising novel agents are currently being tested in the clinical setting of advanced CRPC. These include, androgen receptor inhibitors (MDV3100), androgen biosynthesis inhibitors, angiogenesis inhibitors (thalidomide, lenalidomine, aflibercept, tasquinimod), a novel form of radiotherapy (radium-223), and immune-modulating compounds (PROSTVAC-VF). Improvements in progression-free survival and OS rates, observed with novel agents, in metastatic prostate cancer have led to a shift in treatment paradigm. The challenge will be to position the current established and expected novel treatments in the new landscape of metastatic prostate cancer and to determine at what point and time in the disease course they can best be administered.
The potential for BRAF V600 inhibitors in advanced cutaneous melanoma: rationale and latest evidence
Authors: Lemech Charlotte , Infante Jeffrey , Arkenau Hendrik-Tobias
Published: December 2011

Historically, patients with advanced cutaneous melanoma have a poor prognosis and limited treatment options. The discovery of selective v-raf murine sarcoma viral oncogene homolog B1 (BRAF) V600 mutation as an oncogenic mutation in cutaneous melanoma and the importance of the mitogen-activated protein kinase (MAPK) pathway in its tumourigenesis have changed the treatment paradigm for melanoma. Selective BRAF inhibitors and now MEK inhibitors have demonstrated response rates far higher than standard chemotherapeutic options and we review the phase I–III results for these agents in this article. The understanding of mechanisms of resistance that may occur upstream, downstream, at the BRAF level or bypassing the MAPK pathway provides a platform for rational drug development and combination therapies.
Heads and Tails – Minds and Bodies: Reconsidering the Late Bronze Age Vestby Hoard in Light of Symbolist and Body Perspectives
Authors: Lund Julie , Melheim Lene
Published: December 2011

A pair of animal heads of the Faardal type is central in our reinterpretation of the Late Bronze Age Vestby hoard from eastern Norway. After a period of use and circulation, the heads were mounted on bodies belonging to a different animal species. We argue that the making of the animal figurines and the other high-quality objects of the hoard can only be properly understood against the backdrop of Scandinavian Bronze Age cosmology. This line of thought extends to the presence of a tin bead necklace, which we interpret as a lunar calendar. By combining a ‘body perspective’ – including understandings of body techniques, operational sequences and the ‘sociality’ of objects – with a ‘symbolist perspective’ – including symbol systems, cosmology and intentionality – we put the head back onto the body, so to speak. We also scrutinize the premises for earlier interpretations of the objects' ‘life stories’ and reinterpret their trajectories. This influences the understanding of the act of hoarding, and finally leads to a discussion of how hoarding was also somehow related to the ‘birth’ of the artefacts.
Interactions with the Dead: A Taphonomic Analysis of Burial Practices in Three Megalithic Tombs in County Clare, Ireland
Authors: Beckett Jessica F.
Published: December 2011

Burial is a highly symbolic activity through which concepts of the world are reflected in the representation and treatment of human remains. While mortuary studies in archaeology and anthropology have had a long history, our understanding of Neolithic societies through such analyses is lacking. This article has attempted to broaden our understandings of one such society, focusing upon the megalithic tomb tradition in Ireland, through an integrated study of the burial practices taking place at several sites located on the Burren, County Clare. The Parknabinnia chambered tomb, Poulnabrone portal tomb, and Poulawack Linkardstown-type cairn are located within three kilometres of each other and date to contemporary periods. Several questions are explored through the use of archaeological evidence, osteological analysis, and taphonomy to allow for a broader appreciation of social practices in the past – most notably burial practices. What types of burial practices were taking place; how do the sites compare to each other; and how do they fit within the overall scheme of Neolithic practices we have come to understand?
The Tribal Tattooing of Daunian Women
Authors: Norman Camilla
Published: December 2011

Using cross-cultural ethnographic and archaeological analogy, this paper offers a new interpretation for a specific societal attribute represented on the Iron Age stelae of Daunia, arguing that the designs on the forearms of the female stelae do not represent gloves but are instead tattoos. By questioning a single, long-held, belief about the stelae, it is hoped to highlight a very important principle of method: the need to identify and investigate the socio-cultural context of an image or artefact. For too long the Daunian stelae have been looked at through Hellenized eyes. Unlike other Italic societies of this period, the Daunians appear to have remained reasonably unaffected by Greek and wider Mediterranean influence, suggesting the Iron Age inhabitants of the region retained a strong ethnic identity. With limited evidence of their social and religious life deriving from other sources, any hope of understanding these people properly relies on a correct reading of their stelae, which can only be achieved by firmly placing these monuments within an Adriatic milieu.
Big Enclosures: The Later Neolithic Palisaded Enclosures of Scotland in their Northwestern European Context
Authors: Noble Gordon , Brophy Kenneth
Published: December 2011

Palisaded enclosures were huge enclosed spaces with timber boundaries found across Britain, Ireland, and Scandinavia in the Neolithic. Five such sites have been identified in Scotland dating to the later Neolithic, four of which have been excavated to varying degrees. These sites form the main focus of this paper, which draws in particular on interim results from the authors' excavations at Forteviot, Perth and Kinross, during 2007–2009. The palisaded enclosures of Scotland are part of a wider British and Irish tradition and there are a number of European parallels, the closest of which lie in southern Scandinavia. The palisaded enclosures in Scotland are tightly clustered geographically and chronologically, constructed in the centuries after 2800 cal BC. This paper explores the function, role, and meaning of palisaded enclosures in Scotland and more generally, drawing not just on the architecture of the monuments, but also the individual posts that were used to create the enclosures. The role of these monuments in reconstituting nature is also considered.
Fashioning Identities, Forging Inequalities: Late Neolithic/Copper Age Personal Ornaments of the Portuguese Estremadura
Authors: Thomas Jonathan T.
Published: December 2011

From an analysis of over 3,000 beads and pendants from seven contemporary Late Neolithic/Copper Age (3500–2500 BC) sites in the Portuguese Estremadura, two dominant patterns emerge: (1) most beads show a high degree of standardization in terms of size and shape and are made from local materials; and (2) a minority are made from non-local, rare, and visually distinctive materials (e.g.variscite, ivory), and are less standardized and more labour-intensive. The emphasis on a wide-range of materials suggests that uncommon ornaments may have functioned as ‘value added' materials with special significance, enhancing potential design combinations. Material preferences for beads, bracelets, pendants, plaques, and ground stone tools (da Veiga Ferreira 1951; Lillios 1997, 2008) appear to mirror other Western Mediterranean raw material preferences for ornaments and other polished stone objects (Goñi Quinteiro et al. 1999; Harrison and Orozco Köhler 2001; Pascual Benito 1998; Skeates 2010; Teruel Berbell 1986) suggesting that the Estremadura participated in aspects of a wider system of shared symbolic values.
Horse Burial in First Millennium AD Britain: Issues of Interpretation
Authors: Cross Pamela J.
Published: December 2011

Burial of horses and horse-elements occurred throughout Europe during the first millennium AD. These burials are prevalent in northwest Europe and are perhaps more significant in Britain than previously realised. This article explores the position and value of the horse within Britain during this period and why the burials are likely to represent ritual deposition. Both horse and human-horse burials, are linked to non-Christian burial and sacrificial practices of the Iron Age and Early Medieval period and are particularly associated with Anglo- Saxon and Viking Britain. Some of the traditions appear to reflect the culture described in the Icelandic Sagas, Beowulf, and other legends and chronicles. Archaeologically, the human-horse burials are also linked with high status individuals and ‘warrior graves’, while complete-horse and horse-element burials may represent ritual feasting and/or sacrificial rites which are probably linked with fertility, luck, and the ancestors.
Chronic myeloid leukemia stem cells are not dependent on Bcr-Abl kinase activity for their survival
Authors: Hamilton Ashley , Helgason G. Vignir , Schemionek Mirle , Zhang Bin , Myssina Svetlana , Allan Elaine K. , Nicolini Franck E. , Müller-Tidow Carsten , Bhatia Ravi , Brunton Valerie G. , Koschmieder Steffen , Holyoake Tessa L.
Published: December 2011

Recent evidence suggests chronic myeloid leukemia (CML) stem cells are insensitive to kinase inhibitors and responsible for minimal residual disease in treated patients. We investigated whether CML stem cells, in a transgenic mouse model of CML-like disease or derived from patients, are dependent on Bcr-Abl. In the transgenic model, after retransplantation, donor-derived CML stem cells in which Bcr-Abl expression had been induced and subsequently shut off were able to persist in vivo and reinitiate leukemia in secondary recipients on Bcr-Abl reexpression. Bcr-Abl knockdown in human CD34+ CML cells cultured for 12 days in physiologic growth factors achieved partial inhibition of Bcr-Abl and downstream targets p-CrkL and p-STAT5, inhibition of proliferation and colony forming cells, but no reduction of input cells. The addition of dasatinib further inhibited p-CrkL and p-STAT5, yet only reduced input cells by 50%. Complete growth factor withdrawal plus dasatinib further reduced input cells to 10%; however, the surviving fraction was enriched for primitive leukemic cells capable of growth in a long-term culture-initiating cell assay and expansion on removal of dasatinib and addition of growth factors. Together, these data suggest that CML stem cell survival is Bcr-Abl kinase independent and suggest curative approaches in CML must focus on kinase-independent mechanisms of resistance.
A mutation in the human Uncoordinated 119 gene impairs TCR signaling and is associated with CD4 lymphopenia
Authors: Gorska Magdalena M. , Alam Rafeul
Published: December 2011

Idiopathic CD4 lymphopenia (ICL) is an immunodeficiency disorder of unclear etiology. Here we describe a heterozygous dominant-negative missense mutation (codon 22 GGC→GTC; V22G) of the signaling adaptor protein Uncoordinated 119 (Unc119) in an ICL patient. The patient is a 32-year-old female with < 300 CD4 T cells/μL and with a history of recurrent sinusitis/otitis media, frequent episodes of shingles, a widespread fungal nail infection, fungal dermatitis, oral herpetic lesions, and bronchiolitis obliterans organizing pneumonia after 2 episodes of bacterial pneumonia. The patient's cells have reduced response to TCR stimulation, with impairment in both localization and enzymatic activation of the lymphocyte-specific kinase (Lck) resulting in decreased cell proliferation. Transduction of the mutant Unc119 but not wild-type Unc119 into normal T cells reproduces the signaling and proliferation defects. The mutation disrupts the Unc119-Lck interaction which is normally needed for stimulation of the Lck catalytic activity by TCR. The mutant protein also causes mislocalization of Lck to Rab11+ perinuclear endosomes. The mutation is not present in 2 other patients with ICL, patients with secondary CD4 lymphopenia or 60 healthy subjects. The V22G mutation of Unc119 represents a novel genetic defect in ICL.
The critical role of histone H2A-deubiquitinase Mysm1 in hematopoiesis and lymphocyte differentiation
Authors: Nijnik Anastasia , Clare Simon , Hale Christine , Raisen Claire , McIntyre Rebecca E. , Yusa Kosuke , Everitt Aaron R. , Mottram Lynda , Podrini Christine , Lucas Mark , Estabel Jeanne , Goulding David ,   Sanger Institute Microarray Facility,   Sanger Mouse Genetics Project, Adams Niels , Ramirez-Solis Ramiro , White Jacqui K. , Adams David J. , Hancock Robert E. W. , Dougan Gordon
Published: December 2011

Stem cell differentiation and lineage specification depend on coordinated programs of gene expression, but our knowledge of the chromatin-modifying factors regulating these events remains incomplete. Ubiquitination of histone H2A (H2A-K119u) is a common chromatin modification associated with gene silencing, and controlled by the ubiquitin-ligase polycomb repressor complex 1 (PRC1) and H2A-deubiquitinating enzymes (H2A-DUBs). The roles of H2A-DUBs in mammalian development, stem cells, and hematopoiesis have not been addressed. Here we characterized an H2A-DUB targeted mouse line Mysm1tm1a/tm1a and demonstrated defects in BM hematopoiesis, resulting in lymphopenia, anemia, and thrombocytosis. Development of lymphocytes was impaired from the earliest stages of their differentiation, and there was also a depletion of erythroid cells and a defect in erythroid progenitor function. These phenotypes resulted from a cell-intrinsic requirement for Mysm1 in the BM. Importantly, Mysm1tm1a/tm1a HSCs were functionally impaired, and this was associated with elevated levels of reactive oxygen species, γH2AX DNA damage marker, and p53 protein in the hematopoietic progenitors. Overall, these data establish a role for Mysm1 in the maintenance of BM stem cell function, in the control of oxidative stress and genetic stability in hematopoietic progenitors, and in the development of lymphoid and erythroid lineages.
A phase 1, multicenter, open-label, dose escalation study of elotuzumab in patients with advanced multiple myeloma
Authors: Zonder Jeffrey A. , Mohrbacher Ann F. , Singhal Seema , van Rhee Frits , Bensinger William I. , Ding Han , Fry John , Afar Daniel E. H. , Singhal Anil K.
Published: December 2011

This multicenter, first-in-human study evaluated the safety, tolerability, and pharmacokinetic and pharmacodynamic properties of the anti-CS1 monoclonal antibody elotuzumab. A standard 3 + 3 design was used to determine maximum tolerated dose; dose-limiting toxicities were assessed during cycle 1. Thirty-five patients with relapsed/refractory multiple myeloma were treated with intravenous elotuzumab at doses ranging from 0.5 to 20 mg/kg every 2 weeks. Patients who achieved at least stable disease after 4 treatments could receive another 4 treatments. No maximum tolerated dose was identified up to the maximum planned dose of 20 mg/kg. The most common adverse events, regardless of attribution, were cough, headache, back pain, fever, and chills. Adverse events were generally mild to moderate in severity, and adverse events attributed to study medication were primarily infusion-related. Plasma elotuzumab levels and terminal half-life increased with dose whereas clearance decreased, suggesting target-mediated clearance. CS1 on bone marrow–derived plasma cells was reliably saturated (≥ 95%) at the 10-mg/kg and 20-mg/kg dose levels. Using the European Group for Bone and Marrow Transplantation myeloma response criteria, 9 patients (26.5%) had stable disease. In summary, elotuzumab was generally well tolerated in this population, justifying further exploration of this agent in combination regimens.
IFN-α enhances cross-presentation in human dendritic cells by modulating antigen survival, endocytic routing, and processing
Authors: Spadaro Francesca , Lapenta Caterina , Donati Simona , Abalsamo Laura , Barnaba Vincenzo , Belardelli Filippo , Santini Stefano M. , Ferrantini Maria
Published: December 2011

Cross-presentation allows antigen-presenting cells to present exogenous antigens to CD8+ T cells, playing an essential role in controlling infections and tumor development. IFN-α induces the rapid differentiation of human mono-cytes into dendritic cells, known as IFN-DCs, highly efficient in mediating cross-presentation, as well as the cross-priming of CD8+ T cells. Here, we have investigated the mechanisms underlying the cross-presentation ability of IFN-DCs by studying the intracellular sorting of soluble ovalbumin and nonstructural-3 protein of hepatitis C virus. Our results demonstrate that, independently from the route and mechanism of antigen entry, IFN-DCs are extraordinarily competent in preserving internalized proteins from early degradation and in routing antigens toward the MHC class-I processing pathway, allowing long-lasting, cross-priming capacity. In IFN-DCs, both early and recycling endosomes function as key compartments for the storage of both antigens and MHC-class I molecules and for proteasome- and transporter-associated with Ag processing–dependent auxiliary cross-presentation pathways. Because IFN-DCs closely resemble human DCs naturally occurring in vivo in response to infections and other danger signals, these findings may have important implications for the design of vaccination strategies in neoplastic or chronic infectious diseases.
MicroRNA-27a/b controls endothelial cell repulsion and angiogenesis by targeting semaphorin 6A
Authors: Urbich Carmen , Kaluza David , Frömel Timo , Knau Andrea , Bennewitz Katrin , Boon Reinier A. , Bonauer Angelika , Doebele Carmen , Boeckel Jes-Niels , Hergenreider Eduard , Zeiher Andreas M. , Kroll Jens , Fleming Ingrid , Dimmeler Stefanie
Published: December 2011

MicroRNAs (miRs) are small RNAs that regulate gene expression at the posttranscriptional level. miR-27 is expressed in endothelial cells, but the specific functions of miR-27b and its family member miR-27a are largely unknown. Here we demonstrate that overexpression of miR-27a and miR-27b significantly increased endothelial cell sprouting. Inhibition of both miR-27a and miR-27b impaired endothelial cell sprout formation and induced endothelial cell repulsion in vitro. In vivo, inhibition of miR-27a/b decreased the number of perfused vessels in Matrigel plugs and impaired embryonic vessel formation in zebrafish. Mechanistically, miR-27 regulated the expression of the angiogenesis inhibitor semaphorin 6A (SEMA6A) in vitro and in vivo and targeted the 3′-untranslated region of SEMA6A. Silencing of SEMA6A partially reversed the inhibition of endothelial cell sprouting and abrogated the repulsion of endothelial cells mediated by miR-27a/b inhibition, indicating that SEMA6A is a functionally relevant miR-27 downstream target regulating endothelial cell repulsion. In summary, we show that miR-27a/b promotes angiogenesis by targeting the angiogenesis inhibitor SEMA6A, which controls repulsion of neighboring endothelial cells.
Mouse NK cell–mediated rejection of bone marrow allografts exhibits patterns consistent with Ly49 subset licensing
Authors: Sun Kai , Alvarez Maite , Ames Erik , Barao Isabel , Chen Mingyi , Longo Dan L. , Redelman Doug , Murphy William J.
Published: December 2011

Natural killer (NK) cells can mediate the rejection of bone marrow allografts and exist as subsets based on expression of inhibitory/activating receptors that can bind MHC. In vitro data have shown that NK subsets bearing Ly49 receptors for self-MHC class I have intrinsically higher effector function, supporting the hypothesis that NK cells undergo a host MHC-dependent functional education. These subsets also play a role in bone marrow cell (BMC) allograft rejection. Thus far, little in vivo evidence for this preferential licensing across mouse strains with different MHC haplotypes has been shown. We assessed the intrinsic response potential of the different Ly49+ subsets in BMC rejection by using β2-microglobulin deficient (β2m−/−) mice as donors. Using congenic and allogeneic mice as recipients and depleting the different Ly49 subsets, we found that NK subsets bearing Ly49s, which bind “self-MHC” were found to be the dominant subset responsible for β2m−/− BMC rejection. This provides in vivo evidence for host MHC class I–dependent functional education. Interestingly, all H2d strain mice regardless of background were able to resist significantly greater amounts of β2m−/−, but not wild-type BMC than H2b mice, providing evidence that the rheostat hypothesis regarding Ly49 affinities for MHC and NK-cell function impacts BMC rejection capability.
Normalization of pre-ASCT, FDG-PET imaging with second-line, non–cross-resistant, chemotherapy programs improves event-free survival in patients with Hodgkin lymphoma
Authors: Moskowitz Craig H. , Matasar Matt J. , Zelenetz Andrew D. , Nimer Stephen D. , Gerecitano John , Hamlin Paul , Horwitz Steven , Moskowitz Alison J. , Noy Ariela , Palomba Lia , Perales Miguel-Angel , Portlock Carol , Straus David , Maragulia Jocelyn C. , Schoder Heiko , Yahalom Joachim
Published: December 2011

We previously reported that remission duration < 1 year, extranodal disease, and B symptoms before salvage chemotherapy (SLT) can stratify relapsed or refractory Hodgkin lymphoma (HL) patients into favorable and unfavorable cohorts. In addition, pre-autologous stem cell transplant (ASCT) 18FDG-PET response to SLT predicts outcome. This phase 2 study uses both pre-SLT prognostic factors and post-SLT FDG-PET response in a risk-adapted approach to improve PFS after high-dose radio-chemotherapy (HDT) and ASCT. The first SLT uses 2 cycles of ICE in a standard or augmented dose (ICE/aICE), followed by restaging FDG-PET scan. Patients with a negative scan received a transplant. If the FDG-PET scan remained positive, patients received 4 biweekly doses of gemcitabine, vinorelbine, and liposomal doxorubicin. Patients without evidence of disease progression proceeded to HDT/ASCT; those with progressive disease were study failures. At a median follow-up of 51 months, EFS analyzed by intent to treat as well as for transplanted patients is 70% and 79%, respectively. Patients transplanted with negative FDG-PET, pre-HDT/ASCT after 1 or 2 SLT programs, had an EFS of > 80%, versus 28.6% for patients with a positive scan (P < .001). This prospective study provides evidence that the goal of SLT in patients with Hodgkin lymphoma should be a negative FDG-PET scan before HDT/ASCT. The study was registered at www.clinicaltrials.gov as NCT00255723.
Systemic CD4+ and CD8+ T-cell cytokine profiles correlate with GOLD stage in stable COPD
Authors: Paats Marthe S. , Bergen Ingrid M. , Hoogsteden Henk C. , van der Eerden Menno M. , Hendriks Rudi W.
Published: December 2011

Chronic obstructive pulmonary disease (COPD) is associated with pulmonary and systemic inflammation. Both CD4+ and CD8+ T-lymphocytes play a key role in COPD pathogenesis, but cytokine profiles in circulating T-lymphocytes have not been well characterised.

Here we report the analysis of peripheral blood T-cells from 30 stable COPD patients and 10 healthy never-smokers for interferon (IFN)-γ, interleukin (IL)-4, tumour necrosis factor (TNF)-α and the T-helper 17 cytokines IL-17A, IL-17F and IL-22 by intracellular flow cytometry.

We found significantly increased proportions of IFN-γ+ and TNF-α+ CD8+ T-cells in COPD patients, when compared with healthy controls. This was most evident in patients with less severe disease. In contrast, expression profiles in circulating CD4+ T-cells were similar in COPD patients and healthy controls for all cytokines tested, except for IL-17F. COPD patients with more severely reduced diffusing capacity had lower proportions of IL-17A+ CD4+ T-cells. Proportions of IL-22+ cells in the CD4+ memory T-cell population were significantly increased in active smokers, when compared with past smokers.

Collectively, this comprehensive cytokine analysis of circulating T-cells in COPD patients revealed a correlation for CD8+ T-cells between Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage and IFN-γ or TNF-α expression, but not for CD4+ T-cells.
Interventions to reduce antibiotic prescription for lower respiratory tract infections: Happy Audit study
Authors: Llor Carl , Cots Josep M. , López-Valcárcel Beatriz G. , Arranz Javier , García Guillermo , Ortega Jesús , Gómez Manuel , Guerra Gloria , Monedero María J. , Alcántara Juan D. , Paredes José , Hernández Silvia , Cid Marina , Pérez Carolina , Pineda Vicenta
Published: December 2011

This before–after study aimed to evaluate the effect of two interventions on lowering the prescription of antibiotics in lower respiratory tract infections (LRTI) in Spain.

General practitioners (GPs) registered all cases with LRTIs over 3-week periods before and after an intervention, in 2008 and 2009. Two types of intervention were considered: full-intervention group (FIG), consisting of discussion sessions of the results of the first registry, courses for GPs, guidelines, patient information leaflets, workshops on rapid tests and use of the C-reactive protein (CRP) test; GPs in the partial-intervention group (PIG) underwent all of the above interventions except for the workshop on rapid tests, and they did not have access to CRP. A multilevel logistic regression analysis was performed considering the prescription of an antibiotic as the dependent variable.

210 physicians were assigned to FIG and 70 to PIG. In 2009, 58 new physicians were included as a control group. 5,385 LRTIs were registered. Compared with the control group, the OR of antibiotic prescription after the intervention in the PIG was 0.42 (95% CI: 0.22–0.82) and 0.22 (95% CI: 0.12–0.38) in the FIG.

Intervention led to a reduction in the prescription of antibiotics, mainly when CRP testing was available.
Low affinity PEGylated hemoglobin from Trematomus bernacchii, a model for hemoglobin-based blood substitutes
Authors: Coppola Daniela , Bruno Stefano , Ronda Luca , Viappiani Cristiano , Abbruzzetti Stefania , di Prisco Guido , Verde Cinzia , Mozzarelli Andrea
Published: December 2011

Background

Conjugation of human and animal hemoglobins with polyethylene glycol has been widely explored as a means to develop blood substitutes, a novel pharmaceutical class to be used in surgery or emergency medicine. However, PEGylation of human hemoglobin led to products with significantly different oxygen binding properties with respect to the unmodified tetramer and high NO dioxygenase reactivity, known causes of toxicity. These recent findings call for the biotechnological development of stable, low-affinity PEGylated hemoglobins with low NO dioxygenase reactivity.

Results

To investigate the effects of PEGylation on protein structure and function, we compared the PEGylation products of human hemoglobin and Trematomus bernacchii hemoglobin, a natural variant endowed with a remarkably low oxygen affinity and high tetramer stability. We show that extension arm facilitated PEGylation chemistry based on the reaction of T. bernacchii hemoglobin with 2-iminothiolane and maleimido-functionalyzed polyethylene glycol (MW 5000 Da) leads to a tetraPEGylated product, more homogeneous than the corresponding derivative of human hemoglobin. PEGylated T. bernacchii hemoglobin largely retains the low affinity of the unmodified tetramer, with a p50 50 times higher than PEGylated human hemoglobin. Moreover, it is still sensitive to protons and the allosteric effector ATP, indicating the retention of allosteric regulation. It is also 10-fold less reactive towards nitrogen monoxide than PEGylated human hemoglobin.

Conclusions

These results indicate that PEGylated hemoglobins, provided that a suitable starting hemoglobin variant is chosen, can cover a wide range of oxygen-binding properties, potentially meeting the functional requirements of blood substitutes in terms of oxygen affinity, tetramer stability and NO dioxygenase reactivity.
Signatures of positive selection in Toll-like receptor (TLR) genes in mammals
Authors: Areal Helena , Abrantes Joana , Esteves Pedro J
Published: December 2011

Background

Toll-like receptors (TLRs) are a major class of pattern recognition receptors (PRRs) expressed in the cell surface or membrane compartments of immune and non-immune cells. TLRs are encoded by a multigene family and represent the first line of defense against pathogens by detecting foreigner microbial molecular motifs, the pathogen-associated molecular patterns (PAMPs). TLRs are also important by triggering the adaptive immunity in vertebrates. They are characterized by the presence of leucine-rich repeats (LRRs) in the ectodomain, which are associated with the PAMPs recognition. The direct recognition of different pathogens by TLRs might result in different evolutionary adaptations important to understand the dynamics of the host-pathogen interplay. Ten mammal TLR genes, viral (TLR3, 7, 8, 9) and non-viral (TLR1-6, 10), were selected to identify signatures of positive selection that might have been imposed by interacting pathogens and to clarify if viral and non-viral TLRs might display different patterns of molecular evolution.

Results

By using Maximum Likelihood approaches, evidence of positive selection was found in all the TLRs studied. The number of positively selected codons (PSC) ranged between 2-26 codons (0.25%-2.65%) with the non-viral TLR4 as the receptor with higher percentage of positively selected codons (2.65%), followed by the viral TLR8 (2.50%). The results indicated that viral and non-viral TLRs are similarly under positive selection. Almost all TLRs have at least one PSC located in the LRR ectodomain which underlies the importance of the pathogen recognition by this region.

Conclusions

Our results are not in line with previous studies on primates and birds that identified more codons under positive selection in non-viral TLRs. This might be explained by the fact that both primates and birds are homogeneous groups probably being affected by only a restricted number of related viruses with equivalent motifs to be recognized. The analyses performed in this work encompassed a large number of species covering some of the most representative mammalian groups - Artiodactyla, Rodents, Carnivores, Lagomorphs and Primates - that are affected by different families of viruses. This might explain the role of adaptive evolution in shaping viral TLR genes.
An insight into the sialome of Simulium guianense (DIPTERA:SIMulIIDAE), the main vector of River Blindness Disease in Brazil
Authors: Chagas Andrezza C , Calvo Eric , Pimenta Paulo FP , Ribeiro José MC
Published: December 2011

Background

Little is known about the composition and function of the saliva in black flies such as Simulium guianense, the main vector of river blindness disease in Brazil. The complex salivary potion of hematophagous arthropods counteracts their host's hemostasis, inflammation, and immunity.

Results

Transcriptome analysis revealed ubiquitous salivary protein families--such as the Antigen-5, Yellow, Kunitz domain, and serine proteases--in the S. guianense sialotranscriptome. Insect-specific families were also found. About 63.4% of all secreted products revealed protein families found only in Simulium. Additionally, we found a novel peptide similar to kunitoxin with a structure distantly related to serine protease inhibitors. This study revealed a relative increase of transcripts of the SVEP protein family when compared with Simulium vittatum and S. nigrimanum sialotranscriptomes. We were able to extract coding sequences from 164 proteins associated with blood and sugar feeding, the majority of which were confirmed by proteome analysis.

Conclusions

Our results contribute to understanding the role of Simulium saliva in transmission of Onchocerca volvulus and evolution of salivary proteins in black flies. It also consists of a platform for mining novel anti-hemostatic compounds, vaccine candidates against filariasis, and immuno-epidemiologic markers of vector exposure.
An expression map for Anopheles gambiae
Authors: MacCallum Robert M , Redmond Seth N , Christophides George K
Published: December 2011

Background

Quantitative transcriptome data for the malaria-transmitting mosquito Anopheles gambiae covers a broad range of biological and experimental conditions, including development, blood feeding and infection. Web-based summaries of differential expression for individual genes with respect to these conditions are a useful tool for the biologist, but they lack the context that a visualisation of all genes with respect to all conditions would give. For most organisms, including A. gambiae, such a systems-level view of gene expression is not yet available.

Results

We have clustered microarray-based gene-averaged expression values, available from VectorBase, for 10194 genes over 93 experimental conditions using a self-organizing map. Map regions corresponding to known biological events, such as egg production, are revealed. Many individual gene clusters (nodes) on the map are highly enriched in biological and molecular functions, such as protein synthesis, protein degradation and DNA replication. Gene families, such as odorant binding proteins, can be classified into distinct functional groups based on their expression and evolutionary history. Immunity-related genes are non-randomly distributed in several distinct regions on the map, and are generally distant from genes with house-keeping roles. Each immunity-rich region appears to represent a distinct biological context for pathogen recognition and clearance (e.g. the humoral and gut epithelial responses). Several immunity gene families, such as peptidoglycan recognition proteins (PGRPs) and defensins, appear to be specialised for these distinct roles, while three genes with physically interacting protein products (LRIM1/APL1C/TEP1) are found in close proximity.

Conclusions

The map provides the first genome-scale, multi-experiment overview of gene expression in A. gambiae and should also be useful at the gene-level for investigating potential interactions. A web interface is available through the VectorBase website http://www.vectorbase.org/. It is regularly updated as new experimental data becomes available.
Coupled transcriptome and proteome analysis of human lymphotropic tumor viruses: insights on the detection and discovery of viral genes
Authors: Dresang Lindsay R , Teuton Jeremy R , Feng Huichen , Jacobs Jon M , Camp David G , Purvine Samuel O , Gritsenko Marina A , Li Zhihua , Smith Richard D , Sugden Bill , Moore Patrick S , Chang Yuan
Published: December 2011

Background

Kaposi's sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV) are related human tumor viruses that cause primary effusion lymphomas (PEL) and Burkitt's lymphomas (BL), respectively. Viral genes expressed in naturally-infected cancer cells contribute to disease pathogenesis; knowing which viral genes are expressed is critical in understanding how these viruses cause cancer. To evaluate the expression of viral genes, we used high-resolution separation and mass spectrometry coupled with custom tiling arrays to align the viral proteomes and transcriptomes of three PEL and two BL cell lines under latent and lytic culture conditions.

Results

The majority of viral genes were efficiently detected at the transcript and/or protein level on manipulating the viral life cycle. Overall the correlation of expressed viral proteins and transcripts was highly complementary in both validating and providing orthogonal data with latent/lytic viral gene expression. Our approach also identified novel viral genes in both KSHV and EBV, and extends viral genome annotation. Several previously uncharacterized genes were validated at both transcript and protein levels.

Conclusions

This systems biology approach coupling proteome and transcriptome measurements provides a comprehensive view of viral gene expression that could not have been attained using each methodology independently. Detection of viral proteins in combination with viral transcripts is a potentially powerful method for establishing virus-disease relationships.
Multi-centre evaluation of the speed-oligo Mycobacteria assay for differentiation of Mycobacteriumspp. in clinical isolates
Authors: Hofmann-Thiel Sabine , Turaev Laziz , Alnour Tarig , Drath Lore , Müllerova Maria , Hoffmann Harald
Published: December 2011

Background

A new DNA line probe assay (Speed-oligo Mycobacteria, Vircell) has been launched for rapid differentiation of Mycobacterium spp. from cultures. Compared to other line-probe assays, Speed-oligo Mycobacteria covers a relatively limited spectrum of species but uses a simpler and faster dip-stick technique. The present multi-centre, multi-country study aimed at evaluating the utility and usability of Speed-oligo Mycobacteria in routine mycobacteriology diagnostics. Results from Speed-oligo Myobacteria were compared to those from Genotype CM (HAIN lifescience, Nehren, Germany), another line-probe assay.

Methods

Speed-oligo Mycobacteria assay was performed in three main steps: 1) DNA extraction from cultured material 2) PCR amplification of the target gene and an internal control and 3) hybridization of the PCR products to specific probes by means of a dip-stick.

Results

Two hundred forty-two clinical isolates were recovered from consecutive positive mycobacterial cultures at two German (IML Gauting, Bioscientia Ingelheim), one Czech (KLINLAB Prague), and at a Sudanese (Khartoum) laboratory. All Mycobacterium species covered by the assay were reliably recognized. The rate of false positive results was 1.2% and concerned only the species M. marinum and M. peregrinum. The identification rate, i.e. the proportion of isolates which was correctly differentiated to the level of species or complex by the assay, differed significantly among laboratories being 94.9%, 90.7%, and 75.0% at the study sites IML Gauting, KLINLAB Prague and Bioscientia Ingelheim, respectively. This difference was caused by different spectra of NTM species encountered by the laboratory centres in daily routine diagnostics.

Conclusions

Speed-oligo Mycobacteria assay was proved a rapid and easy-to-perform alternative to conventional line-probe assays. The assay showed excellent sensitivity with regard to identification of genus Mycobacterium and species/complexes covered by the test. However, due to its relatively limited spectrum of taxa, a varying proportion of NTM may not be identified by the assay in daily diagnostics demanding further analyses. The only significant shortcoming in terms of specificity was the misidentification of the clinically relevant species M. marinum.
"Single nucleotide polymorphisms of the OPG/RANKL system genes in primary hyperparathyroidism and their relationship with bone mineral density"
Authors: Piedra María , García-Unzueta María T , Berja Ana , Paule Blanca , Lavín Bernardo A , Valero Carmen , Riancho José A , Amado José A
Published: December 2011

Background

Primary hyperparathyroidism (PHPT) affects mainly cortical bone. It is thought that parathyroid hormone (PTH) indirectly regulates the activity of osteoclasts by means of the osteoprotegerin/ligand of the receptor activator of nuclear factor-κβ (OPG/RANKL) system. Several studies have confirmed thatOPG(osteoprotegerin) andRANKL(ligand of the receptor activator of nuclear factor-κβ) loci are determinants of bone mineral density (BMD) in the general population. The aim of this study is to analyze the relationship between fractures and BMD and the rs3102735 (163 A/G), rs3134070 (245 T/G) and rs2073618 (1181 G/C) SNPs of theOPGand the rs2277438 SNP of theRANKL, in patients with sporadic PHPT.

Methods

We enrolled 298 Caucasian patients with PHPT and 328 healthy volunteers in a cross-sectional study. We analyzed anthropometric data, history of fractures or renal lithiasis, biochemical determinants including markers for bone remodelling, BMD measurements in the lumbar spine, total hip, femoral neck and distal radius, and genotyping for the SNPs to be studied.

Results

Regarding the age of diagnosis, BMI, menopause status, frequency of fractures or renal lithiasis, we found no differences between genotypes in any of the SNPs studied in the PHPT group. Significant lower BMD in the distal radius with similar PTH levels was found in the minor allele homozygotes (GG) compared to heterozygotes and major allele homozygotes in bothOPGrs3102735 (163 A/G) andOPGrs3134070 (245 T/G) SNPs in those with PHPT compared to control subjects. We found no differences between genotypes of theOPGrs2073618 (1181 G/C) SNP with regard to BMD in the PHPT subjects. In the evaluation of rs2277438 SNP of theRANKLin PHPT patients, we found a non significant trend towards lower BMD in the 1/3 distal radius and at total hip in the minor allele homocygotes (GG) genotype group versus heterocygotes and major allele homocygotes (AA).

Conclusions

Our study provides the first evaluation of the relationship between SNPs of theOPG/RANKsystem and sporadic PHPT. Subjects with PHPT and minor homocygote genotype (GG) for theOPGrs3102735 (163 A/G) andOPGrs3134070 (245 T/G) SNPs have lower BMD in the distal radius, and this association does not appear to be mediated by differences in PTH serum levels.
Decreased expression of 17β-hydroxysteroid dehydrogenase type 1 is associated with DNA hypermethylation in colorectal cancer located in the proximal colon
Authors: Rawłuszko Agnieszka Anna , Horbacka Karolina , Krokowicz Piotr , Jagodziński Paweł Piotr
Published: December 2011

Background

The importance of 17β-estradiol (E2) in the prevention of large bowel tumorigenesis has been shown in many epidemiological studies. Extragonadal E2 may form by the aromatase pathway from androstenedione or the sulfatase pathway from estrone (E1) sulfate followed by E1 reduction to E2 by 17-β-hydroxysteroid dehydrogenase (HSD17B1), so HSD17B1 gene expression may play an important role in the production of E2 in peripheral tissue, including the colon.

Methods

HSD17B1 expression was analyzed in colorectal cancer cell lines (HT29, SW707) and primary colonic adenocarcinoma tissues collected from fifty two patients who underwent radical colon surgical resection. Histopathologically unchanged colonic mucosa located at least 10-20 cm away from the cancerous lesions was obtained from the same patients. Expression level of HSD17B1 using quantitative PCR and western blot were evaluated. DNA methylation level in the 5' flanking region of HSD17B1 CpG rich region was assessed using bisulfite DNA sequencing and HRM analysis. The influence of DNA methylation on HSD17B1 expression was further evaluated by ChIP analysis in HT29 and SW707 cell lines. The conversion of estrone (E1) in to E2 was determined by electrochemiluminescence method.

Results

We found a significant decrease in HSD17B1 transcript (p = 0.0016) and protein (p = 0.0028) levels in colorectal cancer (CRC) from the proximal but not distal colon and rectum. This reduced HSD17B1 expression was associated with significantly increased DNA methylation (p = 0.003) in the CpG rich region located in the 5' flanking sequence of the HSD17B1 gene in CRC in the proximal but not distal colon and rectum. We also showed that 5-dAzaC induced demethylation of the 5' flanking region of HSD17B1, leading to increased occupation of the promoter by Polymerase II, and increased transcript and protein levels in HT29 and SW707 CRC cells, which contributed to the increase in E2 formation.

Conclusions

Our results showed that reduced HSD17B1 expression can be associated with DNA methylation in the 5' flanking region of HSD17B1 in CRC from the proximal colon.
Measuring melasma patients' quality of life using willingness to pay and time trade-off methods in thai population
Authors: Leeyaphan Charussri , Wanitphakdeedecha Rungsima , Manuskiatti Woraphong , Kulthanan Kanokvalai
Published: December 2011

Background

Melasma is a common hyperpigmentation disorder that has a significant effect on an individual's quality of life. However, there is no preference-based measurement that reflects quality of life in patients with melasma. The objective of this study was to assess the impact of melasma on quality of life by using a health status measurement - the Dermatology Life Quality Index (DLQI) - and a preference-based measurement - Willingness to Pay (WTP) and Time Trade-Off (TTO).

Methods

A cross-sectional descriptive study was conducted. Seventy-eight patients with melasma who attended the melasma clinic at Siriraj Hospital from February to March 2009 were recruited in this study. The Thai version of the DLQI, questionnaires about WTP, standard TTO, and daily TTO were used to assess patients' quality of life.

Results

Seventy-seven (98.7%) patients were female with a mean age of 47.8 ± 7.9 years. The mean health utility based on standard TTO was 0.96. The utility obtained by the daily TTO method was 0.92 and was significantly correlated with an economically inactive occupation (p < 0.05). The mean monthly WTP for the most effective treatment was 1,157 baht (7.2% of monthly income), ranging from 100 to 5,000 baht (1 USD ~ 35.1 baht). The WTP was significantly correlated with monthly personal income and the total DLQI score.

Conclusion

The WTP method could be a useful tool with which to measure the quality of life of patients with melasma.