bluuebunny/crossref_metadata_2025 huggingface dataset:
Age-specific effects of estrogen receptors' polymorphisms on the bone traits in healthy fertile women: the BONTURNO study
Authors: Massart Francesco , Marini Francesca , Bianchi Gerolamo , Minisola Salvatore , Luisetto Giovanni , Pirazzoli Antonella , Salvi Sara , Micheli Dino , Masi Laura , Brandi Maria Luisa
Published: April 2009

Background

Skeletal characteristics such as height (Ht), bone mineral density (BMD) or bone turnover markers are strongly inherited. Common variants in the genes encoding for estrogen receptor alpha (ESR1) and beta (ESR2) are proposed as candidates for influencing bone phenotypes at the population level.

Methods

We studied 641 healthy premenopausal women aged 20–50 years (yrs) participating into the BONTURNO study. Exclusion criteria were irregular cyclic menses, low trauma fracture, metabolic bone or chronic diseases. Serum C-telopeptide of type I collagen (CTX), osteocalcin (OC), and N-terminal propeptide of type I procollagen (P1NP) were measured in all enrolled subjects, who underwent to lumbar spine (LS), total hip (TH) and femoral neck (FN) BMD evaluation by DXA. Five hundred seventy Caucasian women were genotyped for ESR1 rs2234693 and rs9340799 and ESR2 rs4986938 polymorphisms.

Results

Although no genotype differences were found in body parameters, subjects with combined ESR1 CCGG plus ESR2 AA-AG genotype were taller than those with opposite genotype (P = 0.044). Moreover, ESR1 rs2234693 genotypes correlated with family history of osteoporosis (FHO) and hip fracture (FHF) (P < 0.01), while ESR2 AA-AC genotypes were strongly associated with FHF (OR 2.387, 95% CI 1.432–3.977; P < 0.001).

When clustered by age, 20–30 yrs old subjects, having at least one ESR1 rs2234693 C allele presented lower LS- (P = 0.008) and TH-BMD (P = 0.047) than TT genotypes. In 41–50 yrs age, lower FN-BMD was associated with ESR2 AA (P = 0.0180) subjects than in those with the opposite genotype. ESR1 rs2234693 and rs9340799 and ESR2 rs4986938 polymorphisms did not correlate with age-adjusted values of OC, CTX and P1NP.

Conclusion

These findings support the presence of age-specific effects of ESR1 and ESR2 polymorphisms on various skeletal traits in healthy fertile women.
Scoring mechanisms of p16INK4a immunohistochemistry based on either independent nucleic stain or mixed cytoplasmic with nucleic expression can significantly signal to distinguish between endocervical and endometrial adenocarcinomas in a tissue microarray study
Authors: Koo Chiew-Loon , Kok Lai-Fong , Lee Ming-Yung , Wu Tina S , Cheng Ya-Wen , Hsu Jeng-Dong , Ruan Alexandra , Chao Kuan-Chong , Han Chih-Ping
Published: April 2009

Background

Endocervical adenocarcinomas (ECAs) and endometrial adenocarcinomas (EMAs) are malignancies that affect uterus; however, their biological behaviors are quite different. This distinction has clinical significance, because the appropriate therapy may depend on the site of tumor origin. The purpose of this study is to evaluate 3 different scoring mechanisms of p16INK4a immunohistochemical (IHC) staining in distinguishing between primary ECAs and EMAs.

Methods

A tissue microarray (TMA) was constructed using formalin-fixed, paraffin-embedded tissue from hysterectomy specimens, including 14 ECAs and 24 EMAs. Tissue array sections were immunostained with a commercially available antibody of p16INK4a. Avidin-biotin complex (ABC) method was used for antigens visualization. The staining intensity and area extent of the IHC reactions was evaluated using the semi-quantitative scoring system. The 3 scoring methods were defined on the bases of the following: (1) independent cytoplasmic staining alone (Method C), (2) independent nucleic staining alone (Method N), and (3) mean of the sum of cytoplasmic score plus nucleic score (Method Mean of C plus N).

Results

Of the 3 scoring mechanisms for p16INK4a expression, Method N and Method Mean of C plus N showed significant (p-values < 0.05), but Method C showed non-significant (p = 0.245) frequency differences between ECAs and EMAs. In addition, Method Mean of C plus N had the highest overall accuracy rate (81.6%) for diagnostic distinction among these 3 scoring methods.

Conclusion

According to the data characteristics and test effectiveness in this study, Method N and Method Mean of C plus N can significantly signal to distinguish between ECAs and EMAs; while Method C cannot do. Method Mean of C plus N is the most promising and favorable means among the three scoring mechanisms.
Birth weight and characteristics of endothelial and smooth muscle cell cultures from human umbilical cord vessels
Authors: Martín de Llano José Javier , Fuertes Graciela , Torró Isabel , García Vicent Consuelo , Fayos José Luis , Lurbe Empar
Published: April 2009

Background

Low birth weight has been related to an increased risk for developing high blood pressure in adult life. The molecular and cellular analysis of umbilical cord artery and vein may provide information about the early vascular characteristics of an individual. We have assessed several phenotype characteristics of the four vascular cell types derived from human umbilical cords of newborns with different birth weight. Further follow-up studies could show the association of those vascular properties with infancy and adulthood blood pressure.

Methods

Endothelial and smooth muscle cell cultures were obtained from umbilical cords from two groups of newborns of birth weight less than 2.8 kg or higher than 3.5 kg. The expression of specific endothelial cell markers (von Willebrand factor, CD31, and the binding and internalization of acetylated low-density lipoprotein) and the smooth muscle cell specific α-actin have been evaluated. Cell culture viability, proliferation kinetic, growth fraction (expression of Ki67) and percentage of senescent cells (detection of β-galactosidase activity at pH 6.0) have been determined. Endothelial cell projection area was determined by morphometric analysis of cell cultures after CD31 immunodetection.

Results

The highest variation was found in cell density at the confluence of endothelial cell cultures derived from umbilical cord arteries (66,789 ± 5,093 cells/cm2 vs. 45,630 ± 11,927 cells/cm2, p < 0.05). Morphometric analysis indicated that the projection area of the artery endothelial cells (1,161 ± 198 and 1,544 ± 472 μm2, p < 0.05), but not those derived from the vein from individuals with a birth weight lower than 2.8 kg was lower than that of cells from individuals with a birth weight higher than 3.5 kg.

Conclusion

The analysis of umbilical cord artery endothelial cells, which demonstrated differences in cell size related to birth weight, can provide hints about the cellular and molecular links between lower birth weight and increased adult high blood pressure risk.
Immunomodulatory parasites and toll-like receptor-mediated tumour necrosis factor alpha responsiveness in wild mammals
Authors: Jackson Joseph A , Friberg Ida M , Bolch Luke , Lowe Ann , Ralli Catriona , Harris Philip D , Behnke Jerzy M , Bradley Janette E
Published: April 2009

Background

Immunological analyses of wild populations can increase our understanding of how vertebrate immune systems respond to 'natural' levels of exposure to diverse infections. A major recent advance in immunology has been the recognition of the central role of phylogenetically conserved toll-like receptors in triggering innate immunity and the subsequent recruitment of adaptive response programmes. We studied the cross-sectional associations between individual levels of systemic toll-like receptor-mediated tumour necrosis factor alpha responsiveness and macro- and microparasite infections in a natural wood mouse (Apodemus sylvaticus) population.

Results

Amongst a diverse group of macroparasites, only levels of the nematodeHeligmosomoides polygyrusand the lousePolyplax serratawere correlated (negatively) with innate immune responsiveness (measured by splenocyte tumour necrosis factor alpha responses to a panel of toll-like receptor agonists).Polyplax serratainfection explained a strikingly high proportion of the total variation in innate responses. Contrastingly, faecal oocyst count in microparasiticEimeriaspp. was positively associated with innate immune responsiveness, most significantly for the endosomal receptors TLR7 and TLR9.

Conclusion

Analogy with relevant laboratory models suggests the underlying causality for the observed patterns may be parasite-driven immunomodulatory effects on the host. A subset of immunomodulatory parasite species could thus have a key role in structuring other infections in natural vertebrate populations by affecting the 'upstream' innate mediators, like toll-like receptors, that are important in initiating immunity. Furthermore, the magnitude of the present result suggests that populations free from immunosuppressive parasites may exist at 'unnaturally' elevated levels of innate immune activation, perhaps leading to an increased risk of immunopathology.
Amplitude-modulated electromagnetic fields for the treatment of cancer: Discovery of tumor-specific frequencies and assessment of a novel therapeutic approach
Authors: Barbault Alexandre , Costa Frederico P , Bottger Brad , Munden Reginald F , Bomholt Fin , Kuster Niels , Pasche Boris
Published: April 2009

Purpose

Because in vitro studies suggest that low levels of electromagnetic fields may modify cancer cell growth, we hypothesized that systemic delivery of a combination of tumor-specific frequencies may have a therapeutic effect. We undertook this study to identify tumor-specific frequencies and test the feasibility of administering such frequencies to patients with advanced cancer.

Patients and methods

We examined patients with various types of cancer using a noninvasive biofeedback method to identify tumor-specific frequencies. We offered compassionate treatment to some patients with advanced cancer and limited therapeutic options.

Results

We examined a total of 163 patients with a diagnosis of cancer and identified a total of 1524 frequencies ranging from 0.1 Hz to 114 kHz. Most frequencies (57–92%) were specific for a single tumor type. Compassionate treatment with tumor-specific frequencies was offered to 28 patients. Three patients experienced grade 1 fatigue during or immediately after treatment. There were no NCI grade 2, 3 or 4 toxicities. Thirteen patients were evaluable for response. One patient with hormone-refractory breast cancer metastatic to the adrenal gland and bones had a complete response lasting 11 months. One patient with hormone-refractory breast cancer metastatic to liver and bones had a partial response lasting 13.5 months. Four patients had stable disease lasting for +34.1 months (thyroid cancer metastatic to lung), 5.1 months (non-small cell lung cancer), 4.1 months (pancreatic cancer metastatic to liver) and 4.0 months (leiomyosarcoma metastatic to liver).

Conclusion

Cancer-related frequencies appear to be tumor-specific and treatment with tumor-specific frequencies is feasible, well tolerated and may have biological efficacy in patients with advanced cancer.

Trial registration

clinicaltrials.gov identifier NCT00805337
Pregnancy in the mature adult mouse does not alter the proportion of mammary epithelial stem/progenitor cells
Authors: Britt Kara L , Kendrick Howard , Regan Joseph L , Molyneux Gemma , Magnay Fiona-Ann , Ashworth Alan , Smalley Matthew J
Published: April 2009

Introduction

In humans, an early full-term pregnancy reduces lifetime breast cancer risk by up to 50% whereas a later pregnancy (>35 years old) can increase lifetime risk. Several mechanisms have been suggested, including changes in levels of circulating hormones, changes in the way the breast responds to these hormones, changes in gene expression programmes which may alter susceptibility to transformation and changes to mammary stem cell numbers or behaviour. Previous studies have shown that the mammary tissue isolated from both virgin and parous mice has the ability to repopulate a cleared mammary fat pad in transplant experiments. Limited dilution transplant assays have demonstrated that early pregnancy (at 5 weeks of age) reduces stem/progenitor cell numbers in the mouse mammary epithelium by twofold. However, the effects on stem/progenitor cell numbers in the mammary epithelium of a pregnancy in older animals have not yet been tested.

Methods

Mice were put through a full-term pregnancy at 9 weeks of age, when the mammary epithelium is mature. The total mammary epithelium was purified from parous 7-week post-lactation and age-matched virgin mice and analysed by flow cytometry and limiting dilution cleared fat pad transplants.

Results

There were no significant differences in the proportions of different mammary epithelial cell populations or numbers of CD24+/Low Sca-1- CD49fHigh cells (stem cell enriched basal mammary epithelial compartment). There was no significant difference in stem/progenitor cell frequency based on limiting dilution transplants between the parous and age-matched virgin epithelium.

Conclusions

Although differences between parous and virgin mammary epithelium at later time points post lactation or following multiple pregnancies cannot be ruled out, there are no differences in stem/progenitor cell numbers between mammary epithelium isolated from parous animals which were mated at 9 weeks old and virgin animals. However, a recent report has suggested that animals that were mated at 5 weeks old have a twofold reduction in stem/progenitor cell numbers. This is of interest given the association between early, but not late, pregnancy and breast cancer risk reduction in humans. However, a mechanistic connection between stem cell numbers and breast cancer risk remains to be established.
Remifentanil discontinuation and subsequent intensive care unit-acquired infection: a cohort study
Authors: Nseir Saad , Hoel Jérémy , Grailles Guillaume , Soury-Lavergne Aude , Di Pompeo Christophe , Mathieu Daniel , Durocher Alain
Published: April 2009

Introduction

Recent animal studies demonstrated immunosuppressive effects of opioid withdrawal resulting in a higher risk of infection. The aim of this study was to determine the impact of remifentanil discontinuation on intensive care unit (ICU)-acquired infection.

Methods

This was a prospective observational cohort study performed in a 30-bed medical and surgical university ICU, during a one-year period. All patients hospitalised in the ICU for more than 48 hours were eligible. Sedation was based on a written protocol including remifentanil with or without midazolam. Ramsay score was used to evaluate consciousness. The bedside nurse adjusted sedative infusion to obtain the target Ramsay score. Univariate and multivariate analyses were performed to determine risk factors for ICU-acquired infection.

Results

Five hundred and eighty-seven consecutive patients were included in the study. A microbiologically confirmed ICU-acquired infection was diagnosed in 233 (39%) patients. Incidence rate of ICU-acquired infection was 38 per 1000 ICU-days. Ventilator-associated pneumonia was the most frequently diagnosed ICU-acquired infection (23% of study patients).Pseudomonas aeruginosawas the most frequently isolated microorganism (30%). Multivariate analysis identified remifentanil discontinuation (odds ratio (OR) = 2.53, 95% confidence interval (CI) = 1.28 to 4.99,P= 0.007), simplified acute physiology score II at ICU admission (1.01 per point, 95% CI = 1 to 1.03,P= 0.011), mechanical ventilation (4.49, 95% CI = 1.52 to 13.2,P= 0.006), tracheostomy (2.25, 95% CI = 1.13 to 4.48,P= 0.021), central venous catheter (2.9, 95% CI = 1.08 to 7.74,P= 0.033) and length of hospital stay (1.05 per day, 95% CI = 1.03 to 1.08,P< 0.001) as independent risk factors for ICU-acquired infection.

Conclusions

Remifentanil discontinuation is independently associated with ICU-acquired infection.
Coregulation in human leukocytes of the long pentraxin PTX3 and TSG-6
Authors: Maina Virginia , Cotena Alessia , Doni Andrea , Nebuloni Manuela , Pasqualini Fabio , Milner Caroline M , Day Anthony J , Mantovani Alberto , Garlanda Cecilia
Published: April 2009

Coordinated expression of TSG-6 and PTX3 in myelomonocytic cells and MDDC support the hypothesis that these two proteins may have a role in extracellular matrix remodeling at sites of inflammation.

The prototypic long PTX3 is a multifunctional protein involved in innate resistance to pathogens and in controlling inflammation. TSG-6 is a hyaluronan-binding protein that is involved in ECM remodeling and has anti-inflammatory and chondroprotective functions. PTX3 and TSG-6 are coregulated by growth differentiation factor-9 in granulosa cells, where they are produced during the periovulatory period and play essential roles in the incorporation of hyaluronan into the ECM during cumulus expansion. The present study was designed to assess whether PTX3 and TSG-6 are coregulated in leukocytes, in particular, in phagocytes and DC. Monocytes, macrophages, and myeloid DC were found to produce high levels of TSG-6 and PTX3 in response to proinflammatory mediators (LPS or cytokines). Unstimulated neutrophil polymorphonuclear granulocytes expressed high levels of TSG-6 mRNA, but not PTX3 transcript, and stored both proteins in granules. In contrast, endothelial cells expressed substantial amounts of PTX3 mRNA and low levels of TSG-6 transcript under the conditions tested. Anti-inflammatory cytokines, such as IL-4, dampened LPS-induced TSG-6 and PTX3 expression. Divergent effects were observed with IL-10, which synergizes with TLR-mediated PTX3 induction but inhibits LPS-induced TSG-6 transcription. Immunohistochemical analysis confirms the colocalization of the two proteins in inflammatory infiltrates and in endothelial cells of inflamed tissues. Thus, here we show that myelomonocytic cells and MoDC are a major source of TSG-6 and that PTX3 and TSG-6 are coregulated under most of the conditions tested. The coordinated expression of PTX3 and TSG-6 may play a role in ECM remodeling at sites of inflammation.
Impaired M. tuberculosis-mediated apoptosis in alveolar macrophages from HIV+ persons: potential role of IL-10 and BCL-3
Authors: Patel Naimish R , Swan Katharine , Li Xin , Tachado Souvenir D , Koziel Henry
Published: April 2009

HIV alters the lung environment through increased IL-10, leading to an impaired macrophage apoptotic response to Mycobacterium tuberculosis via upregulation of macrophage BCL-3.

The mechanism of increased MTb disease susceptibility in HIV+ persons remains poorly understood. Apoptosis of macrophages in response to MTb represents a critical host defense response, and decreased apoptosis may represent a mechanism of increased susceptibility to MTb in HIV. In the current study, MTb-mediated apoptosis of human AM was reduced in HIV+ subjects compared with healthy subjects in a TNF-α-dependent manner. IL-10 levels in BALF from HIV+ persons were significantly elevated compared with HIV– persons, and exogenous IL-10 reduced MTb-mediated apoptosis in healthy AM, suggesting that IL-10 could mediate decreased apoptosis observed in HIV. Further study showed that IL-10 reduced TNF release in response to MTb in AM through a reduction in TNF mRNA levels, and exogenous TNF could partially reverse IL-10-associated effects on AM apoptosis. IL-10 did not influence p-IRAK, IκB degradation, or NF-κB p65 nuclear translocation in response to MTb, but IL-10 did increase levels of AM BCL-3, an inhibitor of NF-κB nuclear activity. BCL-3 knockdown in human macrophages increased MTb-mediated TNF release. Importantly, BCL-3 levels in AM from HIV+ subjects were higher compared with healthy subjects. Taken together, these data suggest that elevated lung levels of IL-10 may impair MTb-mediated AM apoptosis in HIV through a BCL-3-dependent mechanism. BCL-3 may represent a potential therapeutic target to treat or prevent MTb disease in HIV+ persons.
The Relationship between Central Adrenal Insufficiency and Sleep-Related Breathing Disorders in Children with Prader-Willi Syndrome
Authors: de Lind van Wijngaarden Roderick F. A. , Joosten Koen F. M. , van den Berg Sandra , Otten Barto J. , de Jong Frank H. , Sweep C. G. J. (Fred) , de Weerd Al W. , Hokken-Koelega Anita C. S.
Published: April 2009

Background: The annual death rate of patients with Prader-Willi syndrome (PWS) is high (3%). Many deaths of children are sudden and unexplained. Sleep apneas have been suggested to play a role in sudden deaths. Recently, we discovered that 60% of patients with PWS suffer from central adrenal insufficiency (CAI) during stress.

Objective: The aim was to study the relationship between CAI and sleep-related breathing disorders.

Design: In 20 children with PWS who underwent a metyrapone test (30 mg/kg at 2330 h), sleep-related breathing was evaluated by polysomnography before the metyrapone test. In addition, we recorded sleep-related breathing in 10 children with PWS during their metyrapone test. CAI was diagnosed when ACTH levels during the metyrapone test were below 33 pmol/liter at 0730 h. All tests were performed during healthy condition.

Setting: The study was conducted in a pediatric intensive care unit and specialized sleep center.

Results: Median (interquartile range) age was 8.4 yr (6.5–10.2). After metyrapone administration, median (interquartile range) central apnea index (number/hour) increased significantly from 2.2 (0.4–4.7) to 5.2 (1.5–7.9) (P = 0.007). The increase tended to be higher in children with CAI [2.8 (2.0–3.9) vs. 1.0 (−0.2 to 2.6); P = 0.09]. During polysomnography before the metyrapone test, sleep-related breathing was worse in children with CAI, who had a significantly higher central apnea index and tended to have a lower minimum oxygen saturation compared to those without CAI (P = 0.03 and P = 0.07).

Conclusions: In children with PWS, the central apnea index increased significantly after metyrapone administration, particularly in those with CAI during stress. In addition, children with CAI had a higher central apnea index compared to those without several months before the metyrapone test.
Hypoglycemia from IGF2 Overexpression Associated with Activation of Fetal Promoters and Loss of Imprinting in a Metastatic Hemangiopericytoma
Authors: Lawson Elizabeth A. , Zhang Xun , Crocker Jonathan T. , Wang Wei-Lien , Klibanski Anne
Published: April 2009

Context: The mechanism of IGF2 overexpression in non-islet-cell tumor hypoglycemia is not understood.

Objective: We investigated the imprinting control and promoter usage for IGF2 expression to identify a mechanism for increased IGF-II production in non-islet-cell tumor hypoglycemia.

Patient and Methods: A patient with metastatic hemangiopericytoma was studied. Tissue from the original hemangiopericytoma, metastatic tumor, and uninvolved liver was analyzed for IGF-II immunohistochemistry. IGF2, a paternally imprinted gene, shares a control region with maternally imprinted H19, a putative tumor suppressor. IGF-II and H19 mRNA expression was compared in metastatic tumor and uninvolved liver by quantitative RT-PCR. Imprinting of IGF2/H19 genes and IGF2 promoter usage in metastatic tumor was investigated by RT-PCR and sequence analysis, and the methylation pattern in the IGF2/H19 imprinting control region was analyzed.

Results: IGF-II protein expression was increased in metastatic tumor vs. uninvolved liver and original tumor. In the metastatic tumor, IGF-II mRNA was increased 60-fold, but H19 mRNA was comparable to uninvolved liver; loss of imprinting of IGF2, but not H19, was identified; no major change in methylation of the IGF2/H19 imprinting control regions was observed; and transcripts from four different IGF2 promoters were detected, compared to two in uninvolved liver.

Conclusions: IGF-2 overexpression, newly acquired in the metastatic tumor, was associated with loss of IGF2 gene imprinting and different promoter usage. The imprinting control mechanism governing the IGF2/H19 locus was intact, as evidenced by normal levels of H19, maintenance of H19 imprinting, and no major change in methylation of the imprinting control regions.
Political Conflict and Power Sharing in the Origins of Modern Colombia
Authors: Mazzuca Sebastián , Robinson James A.
Published: April 2009

After a long period of chaos, political order in Colombia emerged in the mid-1900s. This transition was driven by a change in the institutional allocation of political power. After the Thousand Days’ War, Colombia’s two parties agreed to share power by means of a new set of electoral rules. The incomplete vote, the cornerstone of the new electoral rules, was a strategic concession by the Conservative government to the Liberal opposition. In exchange for permanent representation in the legislatures, Liberals abandoned military insurrection as a political strategy. Transition to proportional representation was completed in 1929 with the introduction of the quotient rule. The quotient rule was also a concession from the government. However, it was not driven by Liberalism’s potential military power but by the institutional power that Liberalism had accumulated since the first concession.
Rethinking the Twelfth-Century Discovery of Nature
Authors: Ritchey Sara
Published: April 2009

According to long-standing scholarly opinion, the twelfth century discovered nature. This essay argues that earthly nature was not discovered in the twelfth century. The twelfth-century authors of the philosophia mundi or the sculptors who fashioned the acanthus capitals at Rheims Cathedral in fact did not think of their work as belonging to the category of nature but to something entirely different from nature–to the order of creation. Continuing to seek “nature” in the medieval past risks overlooking or misunderstanding some profoundly suggestive materials about how people once experienced God, each other, and the world. Examining metaphor and imagery that adopts features of the natural world, this essay thinks through the implications for twelfth-century people's spiritual lives of the idea that God, through the Incarnation, entered not nature, but creation. In particular, the essay examines the role of “trees of incarnation” as contemplative models in women's religious communities for making Christ present in the imagination and in the world. M. D. Chenu's attention to the category of nature in his historical and theological writings is then revisited in order to propose ways of rethinking the manner in which medieval religiosi perceived the material world as a medium for experiencing and continuing the Incarnation of Christ.
Cockfighting as Cultural Allegory in Early Modern England
Authors: Hamill Thomas A.
Published: April 2009

This essay examines the ways in which cockfighting in early modern England operates as an allegorical mode, as a trope for conscribing social relations and phenomena as distinct as male subject-formation and the realization of eschatological truth. Focusing on two distinct cockfighting texts (one that promotes the virtues of cockfighting as a sport and another that figures cockfighting as an extension of animal husbandry), the essay maps the range of cultural and behavioral practices that early modern cockfighting discourse makes possible. Located in allegories of the cockfight are conflicting systems of meaning that at once affirm and disrupt anthropological distinctions between human and animal activity and the anthropocentric ideologies that construct such demarcations. Reading the early modern cockfight challenges us to critique how we engage sport, early modern culture, animals, and allegory itself as signifying texts.
Distinct phases of cardiomyocyte differentiation regulate growth of the zebrafish heart
Authors: de Pater Emma , Clijsters Linda , Marques Sara R. , Lin Yi-Fan , Garavito-Aguilar Zayra V. , Yelon Deborah , Bakkers Jeroen
Published: April 2009

Amongst animal species, there is enormous variation in the size and complexity of the heart, ranging from the simple one-chambered heart of Ciona intestinalis to the complex four-chambered heart of lunged animals. To address possible mechanisms for the evolutionary adaptation of heart size, we studied how growth of the simple two-chambered heart in zebrafish is regulated. Our data show that the embryonic zebrafish heart tube grows by a substantial increase in cardiomyocyte number. Augmented cardiomyocyte differentiation, as opposed to proliferation, is responsible for the observed growth. By using transgenic assays to monitor developmental timing, we visualized for the first time the dynamics of cardiomyocyte differentiation in a vertebrate embryo. Our data identify two previously unrecognized phases of cardiomyocyte differentiation separated in time, space and regulation. During the initial phase, a continuous wave of cardiomyocyte differentiation begins in the ventricle, ends in the atrium, and requires Islet1 for its completion. In the later phase, new cardiomyocytes are added to the arterial pole, and this process requires Fgf signaling. Thus, two separate processes of cardiomyocyte differentiation independently regulate growth of the zebrafish heart. Together, our data support a model in which modified regulation of these distinct phases of cardiomyocyte differentiation has been responsible for the changes in heart size and morphology among vertebrate species.
The mood stabiliser lithium suppresses PIP3 signalling in Dictyostelium and human cells
Authors: King Jason S. , Teo Regina , Ryves Jonathan , Reddy Jonathan V. , Peters Owen , Orabi Ben , Hoeller Oliver , Williams Robin S. B. , Harwood Adrian J.
Published: April 2009

Bipolar mood disorder (manic depression) is a major psychiatric disorder whose molecular origins are unknown. Mood stabilisers offer patients both acute and prophylactic treatment, and experimentally, they provide a means to probe the underlying biology of the disorder. Lithium and other mood stabilisers deplete intracellular inositol and it has been proposed that bipolar mood disorder arises from aberrant inositol (1,4,5)-trisphosphate [IP3, also known as Ins(1,4,5)P3] signalling. However, there is no definitive evidence to support this or any other proposed target; a problem exacerbated by a lack of good cellular models. Phosphatidylinositol (3,4,5)-trisphosphate [PIP3, also known as PtdIns(3,4,5)P3] is a prominent intracellular signal molecule within the central nervous system (CNS) that regulates neuronal survival, connectivity and synaptic function. By using the genetically tractable organism Dictyostelium, we show that lithium suppresses PIP3-mediated signalling. These effects extend to the human neutrophil cell line HL60. Mechanistically, we show that lithium attenuates phosphoinositide synthesis and that its effects can be reversed by overexpression of inositol monophosphatase (IMPase), consistent with the inositol-depletion hypothesis. These results demonstrate a lithium target that is compatible with our current knowledge of the genetic predisposition for bipolar disorder. They also suggest that lithium therapy might be beneficial for other diseases caused by elevated PIP3 signalling.
Filaggrin in the frontline: role in skin barrier function and disease
Authors: Sandilands Aileen , Sutherland Calum , Irvine Alan D. , McLean W. H. Irwin
Published: April 2009

Recently, loss-of-function mutations in FLG, the human gene encoding profilaggrin and filaggrin, have been identified as the cause of the common skin condition ichthyosis vulgaris (which is characterised by dry, scaly skin). These mutations, which are carried by up to 10% of people, also represent a strong genetic predisposing factor for atopic eczema, asthma and allergies. Profilaggrin is the major component of the keratohyalin granules within epidermal granular cells. During epidermal terminal differentiation, the ∼400 kDa profilaggrin polyprotein is dephosphorylated and rapidly cleaved by serine proteases to form monomeric filaggrin (37 kDa), which binds to and condenses the keratin cytoskeleton and thereby contributes to the cell compaction process that is required for squame biogenesis. Within the squames, filaggrin is citrullinated, which promotes its unfolding and further degradation into hygroscopic amino acids, which constitute one element of natural moisturising factor. Loss of profilaggrin or filaggrin leads to a poorly formed stratum corneum (ichthyosis), which is also prone to water loss (xerosis). Recent human genetic studies strongly suggest that perturbation of skin barrier function as a result of reduction or complete loss of filaggrin expression leads to enhanced percutaneous transfer of allergens. Filaggrin is therefore in the frontline of defence, and protects the body from the entry of foreign environmental substances that can otherwise trigger aberrant immune responses.
CTCF and its protein partners: divide and rule?
Authors: Zlatanova Jordanka , Caiafa Paola
Published: April 2009

CTCF is a ubiquitous transcription factor that is involved in numerous, seemingly unrelated functions. These functions include, but are not limited to, positive or negative regulation of transcription, enhancer-blocking activities at developmentally regulated gene clusters and at imprinted loci, and X-chromosome inactivation. Here, we review recent data acquired with state-of-the-art technologies that illuminate possible mechanisms behind the diversity of CTCF functions. CTCF interacts with numerous protein partners, including cohesin, nucleophosmin, PARP1, Yy1 and RNA polymerase II. We propose that CTCF interacts with one or two different partners according to the biological context, applying the Roman principle of governance, `divide and rule' (divide et impera).
Claudin-10 exists in six alternatively spliced isoforms that exhibit distinct localization and function
Authors: Günzel Dorothee , Stuiver Marchel , Kausalya P. Jaya , Haisch Lea , Krug Susanne M. , Rosenthal Rita , Meij Iwan C. , Hunziker Walter , Fromm Michael , Müller Dominik
Published: April 2009

The tight junction protein claudin-10 is known to exist in two isoforms, resulting from two alternative exons, 1a and 1b (Cldn10a, Cldn10b). Here, we identified and characterized another four claudin-10 splice variants in mouse and human. One (Cldn10a_v1) results from an alternative splice donor site, causing a deletion of the last 57 nucleotides of exon 1a. For each of these three variants one further splice variant was identified (Cldn10a_v2, Cldn10a_v3, Cldn10b_v1), lacking exon 4. When transfected into MDCK cells, Cldn10a, Cldn10a_v1 and Cldn10b were inserted into the tight junction, whereas isoforms of splice variants lacking exon 4 were retained in the endoplasmic reticulum. Cldn10a transfection into MDCK cells confirmed the previously described increase in paracellular anion permeability. Cldn10a_v1 transfection had no direct effect, but modulated Cldn10a-induced organic anion permeability. At variance with previous reports in MDCK-II cells, transfection of high-resistance MDCK-C7 cells with Cldn10b dramatically decreased transepithelial resistance, increased cation permeability, and changed monovalent cation selectivity from Eisenman sequence IV to X, indicating the presence of a high field-strength binding site that almost completely removes the hydration shell of the permeating cations. The extent of all these effects strongly depended on the endogenous claudins of the transfected cells.
RalA and the exocyst complex influence neuronal polarity through PAR-3 and aPKC
Authors: Lalli Giovanna
Published: April 2009

Neuronal polarization requires localized cytoskeletal changes and polarized membrane traffic. Here, I report that the small GTPase RalA, previously shown to control neurite branching, also regulates neuronal polarity. RalA depletion, or ectopic expression of constitutively active RalA in cultured neurons inhibit axon formation. However, expression of a constitutively active RalA mutant that is unable to interact with the exocyst complex has no effect on neuronal polarization. Furthermore, depletion of the Sec6, Sec8 or Exo84 subunits of the exocyst complex also leads to unpolarized neurons. Early stages of neuronal polarization are accompanied by increasing levels of interaction of the exocyst complex with PAR-3 and atypical protein kinase C (aPKC), and by the RalA-dependent association of the exocyst complex with PAR-3. Thus, neuronal polarization involves a RalA-regulated association between mediators of vesicle trafficking (exocyst complex) and cell polarity (PAR-3).
Aberrant function and structure of retinal ribbon synapses in the absence of complexin 3 and complexin 4
Authors: Reim Kerstin , Regus-Leidig Hanna , Ammermüller Josef , El-Kordi Ahmed , Radyushkin Konstantin , Ehrenreich Hannelore , Brandstätter Johann Helmut , Brose Nils
Published: April 2009

Complexins regulate the speed and Ca2+ sensitivity of SNARE-mediated synaptic vesicle fusion at conventional synapses. Two of the vertebrate complexins, Cplx3 and Cplx4, are specifically localized to retinal ribbon synapses. To test whether Cplx3 and Cplx4 contribute to the highly efficient transmitter release at ribbon synapses, we studied retina function and structure in Cplx3 and Cplx4 single- and double-knockout mice. Electroretinographic recordings from single and double mutants revealed a cooperative perturbing effect of Cplx3 and Cplx4 deletion on the b-wave amplitude, whereas most other detected effects in both plexiform synaptic layers were additive. Light and electron microscopic analyses uncovered a disorganized outer plexiform layer in the retinae of mice lacking Cplx3 and Cplx4, with a significant proportion of photoreceptor terminals containing spherical free-floating ribbons. These structural and functional aberrations were accompanied by behavioural deficits indicative of a vision deficit. Our results show that Cplx3 and Cplx4 are essential regulators of transmitter release at retinal ribbon synapses. Their loss leads to aberrant adjustment and fine-tuning of transmitter release at the photoreceptor ribbon synapse, alterations in transmission at bipolar cell terminals, changes in the temporal structure of synaptic processing in the inner plexiform layer of the retina and perturbed vision.
Cyclization of 2-Acyl- and 2-Thioacylamino-Benzylcyclopropanes in the Gas Phase and Solution
Authors: Lebedev Albert T. , Giorgi Gianluca , Maloshitskaya Olga A. , Kuchumova Julia , Samgina Tatiana , Dem'yanov Piotr , Karakhanova Nadezhda , Mochalov Sergey , Fedotov Alexandr
Published: April 2009

Mass spectrometry proved itself to be a powerful tool to predict the directions and yields of mono-molecular reactions of organic compounds. Electron ionization (EI) and electrospray ionization (ESI) were used to study possible transformations of N-( ortho-cyclopropylmethylphenyl)arylamides I and N-( ortho-cyclopropylmethylphenyl)arylthioamides II as well as their para-isomers III and IV in a mass spectrometer and to predict directions and yields of their acid catalyzed cyclization reactions. Several five–eight-membered heterocycles were proposed as possible products of intramolecular transformations of compounds I and II. Reactions of compounds I and II in sulfuric acid solutions were carried out and the results obtained were compared with mass spectrometric data. Surprisingly, EI of the studied compounds mimics their solution reactions better than ESI.
How do UK medical graduates rate their knowledge and skills in sexual health and HIV medicine? A national survey
Authors: Estcourt C , Theobald N , Evans D , Lomax N , Copas A , David L , Edwards A , Fisher M
Published: April 2009

The objectives of this study are to determine self-assessed knowledge and skills in sexual health and HIV medicine in preregistration house officers and to explore undergraduate experiences of teaching and assessment in these subjects prior to the launch of National Core Learning Outcomes in Sexual and Reproductive Health and HIV. The study was designed as a postal questionnaire survey. The participants were all UK medical graduates of August 2004. The response rate 1737/4746 (36%). The main outcome measures were Doctors’ views on their preparedness to manage patients with sexual health and HIV-related problems. Since graduation, 90% of respondents had seen at least one patient with a sexually transmitted infection or HIV-related issue. Seventy-six percent felt confident to take a sexual history. In all, 63% and 53% felt competent in male and female genital examination, respectively. Forty-three percent felt they could conduct an appropriate HIV pretest discussion and 59% felt they could recognize clinical indicators suggestive of HIV. Seventy-eight percent had been formally assessed in sexual health and 55% in HIV medicine. Increased confidence in sexual history taking, HIV pretest discussion and recognition of HIV indicators was associated with a longer duration of teaching and formal examination. In conclusion, although the proportion of recent graduates confident in sexual history taking is encouraging, their lack of skill in discussing HIV testing, risk assessment and recognition of possible HIV presentations must be addressed. Integration of National Core Learning Outcomes into all undergraduate curricula is a key step in reducing inconsistencies in undergraduate training.
A survey of Cambodian health-care providers' HIV knowledge, attitudes and intentions to take a sexual history
Authors: Webber G , Edwards N , Graham I D , Amaratunga C , Gaboury I , Keane V , Ros S , McDowell I
Published: April 2009

Cambodia has one of the highest prevalence rates of HIV in Asia and is scaling up HIV testing. We conducted a cross-sectional survey with 358 health care providers in Phnom Penh, Cambodia to assess readiness for voluntary testing and counselling for HIV. We measured HIV knowledge and attitudes, and predictors of intentions to take a sexual history using the Theory of Planned Behaviour. Over 90% of health care providers correctly answered knowledge questions about HIV transmission, but their attitudes were often not positive towards people living with HIV. The Theory of Planned Behaviour constructs explained 56% of the variance in intention to take a sexual history: the control providers perceive they have over taking a sexual history was the strongest contributor (51%), while social pressure explained a further 3%. Attitudes about taking a sexual history did not contribute to intention. Interventions with Cambodian health care providers should focus on improving skills in sexual history-taking.
Concordance between nucleic acid amplification technique and culture for the diagnosis of gonorrhoea
Authors: Creighton S , Revell B , Barrow A
Published: April 2009

The objective of the study was to evaluate the concordance between nucleic acid amplification technique (NAAT) and culture for the diagnosis of Neisseria gonorrhoeae among attendees at a genitourinary medicine clinic in East London. All patients testing positive for N. gonorrhoeae on NAAT and/or culture between 1 April 2007 and 31 August 2008 at the Department of Sexual Health at Homerton University Hospital were included. Male patients had a first void urine sample for NAAT and urethral culture; female patients had a self-taken vulval swab or endocervical sample sent for NAAT and an endocervical culture sample. After interim analysis, discrepant results had both NAAT and culture repeated prior to treatment. Of 159 male patients with a positive NAAT, 22 (13%) had a negative culture. Among 135 female patients with a positive NAAT, 36 (27%) had a negative culture. Three men had a positive culture and negative NAAT. Nineteen of the discrepant samples were retested prior to treatment and 12 (63%) had spontaneously revered to negative. In conclusion, there was concordance in 84% of male and 67% of female samples. In two-thirds of the discrepant cases, the previously positive NAAT had become negative prior to treatment. This study highlights the importance of consideration of the clinical picture when assessing results.
Diagnosis, management and prevalence estimation of gonorrhoea: influences of Aptima Combo 2 assay with alternative target confirmation
Authors: Mahto M , Zia S , Ritchie D , Mallinson H
Published: April 2009

Case-notes and laboratory data were retrospectively reviewed for influences of dual testing by Aptima Combo 2 (AC2) for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) on the diagnosis, management and prevalence estimation of gonorrhoea in the genitourinary (GU) medicine clinic and community. NG positives by AC2 were confirmed by Aptima Gonococcus assay. Unconfirmed positives were rare. Our study showed that in the GU medicine clinic, AC2 detected about 20% extra cases of NG beyond culture. For best standard of care, NG culture and microscopy are still required in some patients to ensure that treatment is rapid and appropriate. Compared to self-referral at the GU medicine clinic, community tests made a substantial contribution to the overall number of NG cases found (40 community versus 35 Macclesfield GU medicine clinic). The ratio of female to male NG cases found was significantly higher ( P = 0.002) in the community (13 M, 27 F) than at the GU medicine clinic (25 M, 10 F). In the community, over 60% of NG infections occurred in chlamydia-negative patients. The overall prevalence of NG in the GU medicine clinic was 1.3%, the true prevalence being much lower at 0.9% on primary test. Prevalence in the community was 0.4%. Data from dual testing in the community can clarify NG prevalence beyond the existing KC60 (sexually transmitted infections) reports.
Increase in clinical prevalence of AIDS implies increase in unsafe medical injections
Authors: Reid S
Published: April 2009

A mass action model developed by the World Health Organization (WHO) estimates that the re-use of contaminated syringes for medical care accounted for 2.5% of HIV infections in sub-Saharan Africa in 2000. The WHO's model applies the population prevalence of HIV infection rather than the clinical prevalence to calculate patients' frequency of exposure to contaminated injections. This approach underestimates iatrogenic exposure risks when progression to advanced HIV disease is widespread. This sensitivity analysis applies the clinical prevalence of HIV to the model and re-evaluates the transmission efficiency of HIV in injections. These adjustments show that no less than 12–17%, and up to 34–47%, of new HIV infections in sub-Saharan Africa may be attributed to medical injections. The present estimates undermine persistent claims that injection safety improvements would have only a minor impact on HIV incidence in Africa.
Threadworm: an infrequent clinical finding in a genitourinary medicine clinic attendee presenting with ano-genital irritation
Authors: Day S
Published: April 2009

A 24-year-old lady presented to an evening genitourinary (GU) clinic with a short history of vulval and anal irritation. On perianal examination, several threadworms were visible. Symptoms resolved with oral mebendazole and strict personal and environmental hygiene. Threadworm is a common and easily treatable cause of pruritis ani, yet is underreported in GU literature. If the history is suggestive, consider performing the diagnostic cellophane test and/or prescribing empirical treatment.
The Influence of Sensitivity for Road Traffic Noise on Residential Location: Does it Trigger a Process of Spatial Selection?
Authors: Nijland Hans A. , Hartemink Sander , van Kamp Irene , van Wee Bert
Published: April 2009

People move to another house for different reasons. It is sometimes presumed that a process of self-selection might take place on the basis of noise sensitivity, i.e., sensitive people would either leave high noise areas or not move into these areas in the first place. Thus, a “survivor population” would remain in the high noise areas. This research aims to investigate whether such a process can be observed in the Netherlands. The study does not show evidence of a process of self-selection based on noise sensitivity. Nevertheless, the results suggest that noise-sensitive people are less satisfied with their living environment and are more willing to move than those who are not noise sensitive. Due to the limited sample size, external validity is limited.