bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Ozone Blood Treatment on the Metabolite Profile of Human Blood
Authors: Travagli Valter , Zanardi Iacopo , Bernini Patrizia , Nepi Stefano , Tenori Leonardo , Bocci Velio
Published: March 2010

Metabonomic characterization of the effects caused by ozone and other stressors on normal human blood was performed. Samples of blood obtained from healthy subjects were treated ex vivo with increasing concentrations of ozone and/or with UV radiation and heat. 1H-NMR analysis of plasma samples after treatments showed the quantitative variation of some metabolites and the formation of new metabolites normally absent. Both the increment of some metabolites like formate, acetoacetate, and acetate and the decrement of pyruvate were of particular interest. Moreover, the oxidation of ascorbic acid and the transformation of uric acid into allantoin after ozonation within the therapeutic concentration range were observed. In the ozonated spectra, 2 unidentified peaks appeared at 2.82 ppm and 8.08 ppm. They are related to the direct antioxidant activity of albumin in the presence of ozone and they could be considered as specific markers of the blood ozonation.
Critical action learning, policy learning and small firms: An inquiry
Authors: Ram Monder , Trehan Kiran
Published: March 2010

This article presents a conceptual and empirical synthesis of ‘critical action learning’ (CAL) and ‘policy learning’ (PL). We pursue this undertaking by reflecting upon a five-year inquiry into an initiative that aimed to provide business support to an action learning set comprising eight African-Caribbean entrepreneurs. CAL and PL share commitments that are riven with tensions, which we explore in our inquiry in relation to three themes that arose during the course of the investigation. The findings demonstrate how a synthesis of CAL and PL can: enrich CAL by recognizing the centrality of emotional and power relations; provide a vehicle to examine the tensions and dynamics that attend policy implementation; illustrate the merits of an experiential approach to evaluation. We contribute to debates on criticality in action learning and the need for more grounded approaches to the evaluation of initiatives directed at small firms.
Fighting the Last War
Authors: Needham Catherine
Published: March 2010

The concept of policy transfer (or ‘lesson-drawing’) has been used by theorists of public policy to explain how programmes developed in one context emerge in another. This article explores how far the insights of policy transfer can be used to understand better how campaign strategies move cross-nationally in cases where lender and borrower campaigns can be identified. Drawing on the policy transfer literature, it puts forward the following hypotheses: (1) transfers are from incumbent to opposition parties; (2) transfers are between parties with similar ideological positions; (3) the US is the dominant lender country; (4) party leaders, party staff and consultants are the agents of transfer; and (5) campaign strategy transfer is increasing over time. Based on a set of 32 election campaigns from the UK, USA, Israel and Germany, the article finds high levels of support for hypotheses 2—4 but more limited support for hypotheses 1 and 5. Like policy transfer, campaign strategy transfer involves a range of personnel from like-minded parties, with the United States as the predominant lender. However, campaigners appear more willing than policy-makers to learn ‘negative lessons’ (Dolowitz and Marsh, 2000: 12), and there is no evidence that campaign strategy transfer is becoming more prevalent over time.
Adhesive Layer Effects on PZT-induced Lamb Waves at Elevated Temperatures
Authors: Ha Sungwon , Lonkar Kuldeep , Mittal Amrita , Chang Fu-Kuo
Published: March 2010

The role of the adhesive layer on PZT-induced Lamb wave propagation in structures exposed to elevated temperatures is presented in this article. Both experiments and numerical simulations were performed to study the effects of the adhesive layer on sensor signal at elevated temperatures. Experimentally, signals from PZT transducers with different adhesive thicknesses (40 and 120 μm) were investigated up to 500 kHz. In model simulations, the spectral element package (PESEA), which was developed previously, was adopted to simulate the test results. The simulations agreed with the experimental data quite well. Parametric studies were was then performed using PESEA to evaluate the effect of adhesive layer on PZT-induced Lamb wave propagation at elevated temperatures as compared to other mechanical properties of the host structure and PZT materials; these studies revealed that the stiffness change of adhesive layer due to temperature is the most influential parameter for the change in sensor signals as compared to other mechanical properties, and that the thickness of the adhesive layer can affect a sensor signal in a different manner at elevated temperatures. This study shows PESEA can reasonably simulate the adhesive layer effect at elevated temperatures and hence can be a useful tool for understanding the behavior of Lamb wave propagation generated by adhesively bonded PZTs on structures.
Identification of Structural Damage in a Vehicular Bridge using Artificial Neural Networks
Authors: González-Pérez C. , Valdés-González J.
Published: March 2010

This article presents the application of artificial neural networks (ANNs) for the structural damage detection to bending in the girders of a vehicular bridge. An analytical model of the bridge was developed to generate 12,800 damage scenarios, in which the flexural stiffness of the elements was modified to simulate the damage. Such rigidities were used as output data for the network, while the modal strain energy differences were used as input data. To verify the NNs generalization capability in presence of noise in the measurements, four levels of noise were analyzed (2.5%, 5.0%, 7.5%, and 10.0%). It was observed that the developed NNs model is able to predict with high accuracy the location and severity of the damage in the studied bridge.
Media Uses and Effects in New Democracies: The Case of Poland’s 2005 Parliamentary and Presidential Elections
Authors: Tworzecki Hubert , Semetko Holli A.
Published: March 2010

To what extent are established democracies and new democracies moving closer together in terms of the impact of media messages in electoral campaigns? Drawing on a content analysis of main evening television news on flagship programs over two months during the Polish National Election Study’s two-wave panel study ( N = 1,200) spanning parliamentary and presidential elections in September and October 2005, the authors identify the impact of media use on political knowledge, political participation, and political learning. They demonstrate the positive effect of media use on political knowledge, on participation in a variety of campaign activities other than voting, and on the stabilization of issue positions. Findings support those from recent elections in established democracies. The study suggests that in the context of more highly volatile electorates in the new democracies of East-Central Europe, campaign information in the news media plays a critical role in campaigns and electoral outcomes.
Phenotypic effects of the circadian gene Cryptochrome 2 on cancer-related pathways
Authors: Hoffman Aaron E , Zheng Tongzhang , Ba Yue , Stevens Richard G , Yi Chun-Hui , Leaderer Derek , Zhu Yong
Published: March 2010

Background

Circadian genes continue to gain attention as important transcriptional regulators with the potential to influence a variety of biological pathways, including many cancer-related processes. The core circadian gene cryptochrome 2 (CRY2) is essential for proper circadian timing, and is a key component of the negative arm of the circadian feedback loop. As such, aberrant expression of CRY2 may influence carcinogenic processes and thereby impact cancer susceptibility.

Methods

We silenced CRY2 in breast cancer cell lines (MCF-7) using small-interfering oligos (siRNA) and measured the impact of CRY2 knockdown on a number of cancer-relevant parameters. Cell cycle distribution, cell viability, and apoptotic response were measured in CRY2 knockdown (CRY2-) and normal (CRY2+) cell populations using flow cytometry in cells with and without exposure to a mutagen challenge. DNA damage accumulation was measured using the single cell gel electrophoresis (comet) assay, and damage was quantified using the Olive tail moment, which considers the amount and distance of DNA migration away from the nucleus, indicative of DNA strand breaks. Expression changes in cancer-relevant transcripts were measured by whole genome microarray. The Student's t-test was used for statistical comparisons, and P-values obtained from the microarray were adjusted for multiple comparisons using the false discovery rate correction, in order to obtain an adjusted Q-value for each observation.

Results

The comet assay results indicated that upon exposure to the same dose of chemical mutagen, CRY2- cells accumulate significantly more unrepaired DNA damage than CRY2+ cells (P = 0.040), suggesting that CRY2 may be important for DNA repair. In addition, a number of transcripts with relevance for DNA damage repair displayed altered expression following CRY2 silencing. These included BCCIP (Q = 0.002), BCL2 (Q = 0.049), CCND1 (Q = 0.009), CDKN1A (Q < 0.001), GADD45A (Q = 0.002), HERC5 (Q < 0.001), MCM5 (Q = 0.042), PPP1R15A (Q < 0.001), SUMO1 (Q < 0.001), and UBA1 (Q = 0.023). However, no significant influence of CRY2 knockdown on cell cycle distributions, cell cycle checkpoints in response to mutagen challenge, or apoptotic response was detected.

Conclusions

In total, these data suggest a limited, but potentially important role for CRY2 in the regulation of DNA damage repair and the maintenance of genomic stability. Future investigations may focus on identifying the mechanisms by which CRY2 may regulate the expression of transcripts with known relevance for carcinogenesis.
Functional significance of the hepaCAM gene in bladder cancer
Authors: He Yunfeng , Wu Xiaohou , Luo Chunli , Wang Lie , Lin Jie
Published: March 2010

Background

The hepaCAM gene encodes a new immunoglobulin-like cell adhesion molecule, and its expression is suppressed in a variety of human cancers. Additionally, hepaCAM possesses properties often observed in tumor suppressor genes. However, the expression and biological function of hepaCAM has not been investigated in bladder cancer. Therefore we sought to examine hepaCAM expression and the relationship between its structure and function in human transitional cell carcinoma of bladder (TCCB).

Materials and methods

HepaCAM expression was evaluated in 28 normal and 34 TCCB bladder specimens and 2 TCCB cell lines using semi-quantitative RT-PCR. The wild-type hepaCAM and the extracellular domain-truncated mutant gene were transfected into the TCCB cell line T24, and the biological properties of both the wild-type gene and the domain-truncated mutant were then assessed.

Results

HepaCAM expression was down-regulated in 82% (28/34) of TCCB specimens and undetectable in the 2 TCCB cell lines tested. The localization of hepaCAM appeared to be dependent on cell density in T24 cells. In widely spread cells, hepaCAM accumulated on the perinuclear membrane and the cell surface protrusions, whereas in confluent cells, hepaCAM was predominantly localized at the sites of cell-cell contacts on the cell membrane. Functionally, hepaCAM expressed not only increased cell spreading, delayed cell detachment, enhanced wound healing and increased cell invasion; it also inhibited cell growth (P < 0.01). When the extracellular domain was deleted, the localization of hepaCAM was significantly altered, and it lost both its adhesive function and its influence on cell growth.

Conclusions

HepaCAM is involved in cell adhesion and growth control, and its expression is frequently silenced in TCCB. The extracellular domain of hepaCAM is essential to its physiological and biological functions.
Effect of an individually tailored one-year energy balance programme on body weight, body composition and lifestyle in recent retirees: a cluster randomised controlled trial
Authors: Werkman Andrea , Hulshof Paul JM , Stafleu Annette , Kremers Stef PJ , Kok Frans J , Schouten Evert G , Schuit Albertine J
Published: March 2010

Background

The increased prevalence of overweight and obesity warrants preventive actions, particularly among people in transitional stages associated with lifestyle changes, such as occupational retirement. The purpose is to investigate the effect of a one year low-intensity computer-tailored energy balance programme among recent retirees on waist circumference, body weight and body composition, blood pressure, physical activity and dietary intake.

Methods

A randomised controlled trial was conducted among recent retirees (N = 413; mean age 59.5 years). Outcome measures were assessed using anthropometry, bio-impedance, blood pressure measurement and questionnaires.

Results

Waist circumference, body weight and blood pressure decreased significantly in men of the intervention and control group, but no significant between-group-differences were observed at 12 or at 24-months follow-up. A significant effect of the programme was only observed on waist circumference (-1.56 cm (95%CI: -2.91 to -0.21)) at 12 month follow up among men with low education (n = 85). Physical activity and dietary behaviours improved in both the intervention and control group during the intervention period. Although, these behaviours changed more favourably in the intervention group, these between-group-differences were not statistically significant.

Conclusions

The multifaceted computer-tailored programme for recent retirees did not appear to be effective. Apparently the transition to occupational retirement and/or participation in the study had a greater impact than the intervention programme.

Trial registration

Clinical Trials NCT00122213.
Factors associated with educational aspirations among adolescents: cues to counteract socioeconomic differences?
Authors: Madarasova Geckova Andrea , Tavel Peter , van Dijk Jitse P , Abel Thomas , Reijneveld Sijmen A
Published: March 2010

Background

Our study aims to follow this effort and to explore the association between health, socioeconomic background, school-related factors, social support and adolescents' sense of coherence and educational aspirations among adolescents from different educational tracks and to contribute to the existing body of knowledge on the role of educational aspirations in the social reproduction of health inequalities. We expect that socioeconomic background will contribute to the development of educational aspirations, but this association will be modified by available social and individual resources, which may be particularly favourable for the group of adolescents who are on lower educational tracks, since for them such resources may lead to gaining a higher educational level.

Methods

We collected data on the socioeconomic background (mother's and father's education and employment status, doubts about affordability of future study), school-related factors (school atmosphere, school conditions, attitudes towards school), perceived social support, sense of coherence (manageability, comprehensibility, meaningfulness) and the self-rated health of a national sample of Slovak adolescents (n = 1992, 53.5% females, mean age 16.9 years). We assessed the association of these factors with educational aspirations, overall and by educational tracks (grammar schools, specialised secondary schools, vocational schools).

Results

We found statistically significant associations with educational aspirations for the factors parental educational level, father's unemployment, doubts about the affordability of future study, school atmosphere, attitude towards school, social support from the father and a sense of coherence. Social support from the mother and friends was not associated with educational aspiration, nor was self-rated health. Besides affinity towards school, the determinants of educational aspirations differed among adolescents on different educational tracks. Educational aspirations of grammar school students were associated with father's education, while the aspirations of their peers on lower educational tracks had a stronger association with mother's education and perceived social support from father and friends. Moreover, a sense of coherence contributes to the reporting of educational aspiration by students on different educational tracks.

Conclusions

Characteristics of the school environment, the family and the individual adolescent are all associated with the level of educational aspiration, but in a different way for different educational tracks. Interventions aimed at reducing socioeconomic inequalities in health via the educational system should, therefore, take this variation and the rather pivotal role of the father into account.
Different stress-related phenotypes of BALB/c mice from in-house or vendor: alterations of the sympathetic and HPA axis responsiveness
Authors: Olfe Jakob , Domanska Grazyna , Schuett Christine , Kiank Cornelia
Published: March 2010

Background

Laboratory routine procedures such as handling, injection, gavage or transportation are stressful events which may influence physiological parameters of laboratory animals and may interfere with the interpretation of the experimental results. Here, we investigated if female BALB/c mice derived from in-house breeding and BALB/c mice from a vendor which were shipped during their juvenile life differ in their HPA axis activity and stress responsiveness in adulthood.

Results

We show that already transferring the home cage to another room is a stressful event which causes an increased HPA axis activation for at least 24 hours as well as a loss of circulating lymphocytes which normalizes during a few days after transportation. However and important for the interpretation of experimental data, commercially available strain-, age- and gender-matched animals that were shipped over-night showed elevated glucocorticoid levels for up to three weeks after shipment, indicating a heightened HPA axis activation and they gained less body weight during adolescence. Four weeks after shipment, these vendor-derived mice showed increased corticosterone levels at 45-min after intraperitoneal ACTH challenge but, unexpectedly, no acute stress-induced glucocorticoid release. Surprisingly, activation of monoaminergic pathways were identified to inhibit the central nervous HPA axis activation in the vendor-derived, shipped animals since depletion of monoamines by reserpine treatment could restore the stress-induced HPA axis response during acute stress.

Conclusions

In-house bred and vendor-derived BALB/c mice show a different stress-induced HPA axis response in adulthood which seems to be associated with different central monoaminergic pathway activity. The stress of shipment itself and/or differences in raising conditions, therefore, can cause the development of different stress response phenotypes which needs to be taken into account when interpreting experimental data.
Influence of production process design on inclusion bodies protein: the case of an Antarctic flavohemoglobin
Authors: Parrilli Ermenegilda , Giuliani Maria , Marino Gennaro , Tutino Maria Luisa
Published: March 2010

Background

Protein over-production in Escherichia coli often results in formation of inclusion bodies (IBs). Some recent reports have shown that the aggregation into IBs does not necessarily mean that the target protein is inactivated and that IBs may contain a high proportion of correctly folded protein. This proportion is variable depending on the protein itself, the genetic background of the producing cells and the expression temperature. In this paper we have evaluated the influence of other production process parameters on the quality of an inclusion bodies protein.

Results

The present paper describes the recombinant production in Escherichia coli of the flavohemoglobin from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. Flavohemoglobins are multidomain proteins requiring FAD and heme cofactors. The production was carried out in several different experimental setups differing in bioreactor geometry, oxygen supply and the presence of a nitrosating compound. In all production processes, the recombinant protein accumulates in IBs, from which it was solubilized in non-denaturing conditions. Comparing structural properties of the solubilized flavohemoglobins, i.e. deriving from the different process designs, our data demonstrated that the protein preparations differ significantly in the presence of cofactors (heme and FAD) and as far as their secondary and tertiary structure content is concerned.

Conclusions

Data reported in this paper demonstrate that other production process parameters, besides growth temperature, can influence the structure of a recombinant product that accumulates in IBs. To the best of our knowledge, this is the first reported example in which the structural properties of a protein solubilized from inclusion bodies have been correlated to the production process design.
Inhibition of human immunodeficiency virus type-1 by cdk inhibitors
Authors: Guendel Irene , Agbottah Emmanuel T , Kehn-Hall Kylene , Kashanchi Fatah
Published: March 2010

Current therapy for human immunodeficiency virus (HIV-1) infection relies primarily on the administration of anti-retroviral nucleoside analogues, either alone or in combination with HIV-protease inhibitors. Although these drugs have a clinical benefit, continuous therapy with the drugs leads to drug-resistant strains of the virus. Recently, significant progress has been made towards the development of natural and synthetic agents that can directly inhibit HIV-1 replication or its essential enzymes. We previously reported on the pharmacological cyclin-dependent kinase inhibitor (PCI) r-roscovitine as a potential inhibitor of HIV-1 replication. PCIs are among the most promising novel antiviral agents to emerge over the past few years. Potent activity on viral replication combined with proliferation inhibition without the emergence of resistant viruses, which are normally observed in HAART patients; make PCIs ideal candidates for HIV-1 inhibition. To this end we evaluated twenty four cdk inhibitors for their effect on HIV-1 replication in vitro. Screening of these compounds identified alsterpaullone as the most potent inhibitor of HIV-1 with activity at 150 nM. We found that alsterpaullone effectively inhibits cdk2 activity in HIV-1 infected cells with a low IC50 compared to control uninfected cells. The effects of alsterpaullone were associated with suppression of cdk2 and cyclin expression. Combining both alsterpaullone and r-roscovitine (cyc202) in treatment exhibited even stronger inhibitory activities in HIV-1 infected PBMCs.
Histone H1 variant-specific lysine methylation by G9a/KMT1C and Glp1/KMT1D
Authors: Weiss Thomas , Hergeth Sonja , Zeissler Ulrike , Izzo Annalisa , Tropberger Philipp , Zee Barry M , Dundr Miroslav , Garcia Benjamin A , Daujat Sylvain , Schneider Robert
Published: March 2010

Background

The linker histone H1 has a key role in establishing and maintaining higher order chromatin structure and in regulating gene expression. Mammals express up to 11 different H1 variants, with H1.2 and H1.4 being the predominant ones in most somatic cells. Like core histones, H1 has high levels of covalent modifications; however, the full set of modifications and their biological role are largely unknown.

Results

In this study, we used a candidate screen to identify enzymes that methylate H1 and to map their corresponding methylation sites. We found that the histone lysine methyltransferases G9a/KMT1C and Glp1/KMT1D methylate H1.2 in vitro and in vivo, and we mapped this novel site to lysine 187 (H1.2K187) in the C-terminus of H1. This H1.2K187 methylation is variant-specific. The main target for methylation by G9a in H1.2, H1.3, H1.5 and H1.0 is in the C-terminus, whereas H1.4 is preferentially methylated at K26 (H1.4K26me) in the N-terminus. We found that the readout of these marks is different; H1.4K26me can recruit HP1, but H1.2K187me cannot. Likewise, JMJD2D/KDM4 only reverses H1.4K26 methylation, clearly distinguishing these two methylation sites. Further, in contrast to C-terminal H1 phosphorylation, H1.2K187 methylation level is steady throughout the cell cycle.

Conclusions

We have characterised a novel methylation site in the C-terminus of H1 that is the target of G9a/Glp1 both in vitro and in vivo. To our knowledge, this is the first demonstration of variant-specific histone methylation by the same methyltransferases, but with differing downstream readers, thereby supporting the hypothesis of H1 variants having specific functions.
Myofiber stress-response in myositis: parallel investigations on patients and experimental animal models of muscle regeneration and systemic inflammation
Authors: Vitadello Maurizio , Doria Andrea , Tarricone Elena , Ghirardello Anna , Gorza Luisa
Published: March 2010

Introduction

The endoplasmic reticulum (ER) stress-response, evoked in mice by the overexpression of class I major histocompatibility complex antigen (MHC-I), was proposed as a major mechanism responsible for skeletal muscle damage and dysfunction in autoimmune myositis. The present study was undertaken to characterize in more detail the ER stress-response occurring in myofibers of patients with inflammatory myopathies, focusing on the expression and distribution of Grp94, calreticulin and Grp75, three ER chaperones involved in immunomodulation.

Methods

Muscle biopsies were obtained from seven healthy subjects and 29 myositis patients, who were subdivided into groups based on the morphological evidence of inflammation and/or sarcolemmal immunoreactivity for MHC-I. Biopsies were analyzed by means of immunohistochemistry and western blot using anti-Grp94, anti-calreticulin and anti-Grp75 specific antibodies. Parallel analyses on these ER chaperones were conducted in rabbit and/or murine skeletal muscle after experimental induction of regeneration or systemic inflammation.

Results

Upregulation of Grp94 characterized regenerating myofibers of myositis patients (P = 0.03, compared with values detected in biopsies without signs of muscle regeneration) and developing and regenerating myofibers of mouse muscles. Conversely, levels of calreticulin and Grp75 increased about fourfold and twofold, respectively, in patient biopsies positive for sarcolemmal MHC-I immunoreactivity, compared with healthy subjects and patients negative for both inflammation and MHC-I labeling (P < 0.005). Differently from calreticulin, the Grp75 level increased significantly also in patient biopsies that displayed occasional sarcolemmal MHC-I immunoreactivity (P = 0.002), suggesting the interference of other mechanisms. Experimental systemic inflammation achieved in mice and rabbits by a single injection of bacterial lipopolysaccharide significantly increased Grp75 and calreticulin but not MHC-I expression in muscles.

Conclusions

These results indicate that, in myositis patients, muscle regeneration and inflammation, in addition to MHC-I upregulation, do evoke an ER stress-response characterized by the increased expression of Grp94 and Grp75, respectively. The increase in the muscle Grp75 level in patients showing occasional immunoreactivity for sarcolemmal MHC-I might be considered further as a broader indicator of idiopathic inflammatory myopathy.
Circulating Secretin Activates Supraoptic Nucleus Oxytocin and Vasopressin Neurons via Noradrenergic Pathways in the Rat
Authors: Velmurugan Sathya , Brunton Paula J. , Leng Gareth , Russell John A.
Published: March 2010

Secretin is a 27-amino acid brain-gut peptide from duodenal S-cells. We tested the effects of systemic administration of secretin to simulate its postprandial release on neuroendocrine neurons of the supraoptic nucleus (SON) in urethane-anesthetized female rats. Secretin dose-dependently increased the firing rate of oxytocin neurons, more potently than cholecystokinin, and dose-dependently increased plasma oxytocin concentration. The effect of secretin on SON vasopressin neurons was also predominantly excitatory, in contrast to the inhibitory actions of cholecystokinin. To explore the involvement of noradrenergic inputs in secretin-induced excitation, benoxathian, an α1-adrenoceptor antagonist, was infused intracerebroventricularly. Benoxathian intracerebroventricular infusion blocked the excitation by secretin of both oxytocin and vasopressin neurons. To test the role of local noradrenaline release in the SON, benoxathian was microdialyzed onto the SON. The basal firing rate of oxytocin neurons was slightly reduced and the secretin-induced excitation was attenuated during benoxathian microdialysis. Hence, noradrenergic pathways mediate the excitation by systemic secretin of oxytocin neurons via α1-adrenoceptors in the SON. As both systemic secretin and oxytocin are involved in regulating gastrointestinal functions and natriuresis, systemically released secretin might act partly through oxytocin.
Plasma 25-Hydroxyvitamin D Levels and Fracture Risk in a Community-Based Cohort of Elderly Men in Sweden
Authors: Melhus Håkan , Snellman Greta , Gedeborg Rolf , Byberg Liisa , Berglund Lars , Mallmin Hans , Hellman Per , Blomhoff Rune , Hagström Emil , Ärnlöv Johan , Michaëlsson Karl
Published: March 2010

Context: Blood levels of 25-hydroxyvitamin D [25(OH)D] is the generally accepted indicator of vitamin D status, but no universal reference level has been reached.

Objective: The objective of the study was to determine the threshold at which low plasma 25(OH)D levels are associated with fractures in elderly men and clarify the importance of low levels on total fracture burden.

Design and Participants: In the Uppsala Longitudinal Study of Adult Men, a population-based cohort (mean age, 71 yr, n = 1194), we examined the relationship between 25(OH)D and risk for fracture. Plasma 25(OH)D levels were measured with high-pressure liquid chromatography-mass spectrometry.

Setting: The study was conducted in the municipality of Uppsala in Sweden, a country with a high fracture incidence.

Main Outcome Measure: Time to fracture was measured.

Results: During follow-up (median 11 yr), 309 of the participants (26%) sustained a fracture. 25(OH)D levels below 40 nmol/liter, which corresponded to the fifth percentile of 25(OH)D, were associated with a modestly increased risk for fracture, multivariable-adjusted hazard ratio 1.65 (95% confidence interval 1.09–2.49). No risk difference was detected above this level. Approximately 3% of the fractures were attributable to low 25(OH)D levels in this population.

Conclusions: Vitamin D insufficiency is not a major cause of fractures in community-dwelling elderly men in Sweden. Despite the fact that cutaneous synthesis of previtamin D during the winter season is undetectable at this northern latitude of 60°, only one in 20 had 25(OH)D levels below 40 nmol/liter, the threshold at which the risk for fracture started to increase. Genetic adaptations to limited UV light may be an explanation for our findings.
Long-Term Efficacy of Lymph Node Reoperation for Persistent Papillary Thyroid Cancer
Authors: Al-Saif Osama , Farrar William B. , Bloomston Mark , Porter Kyle , Ringel Matthew D. , Kloos Richard T.
Published: March 2010

Objective: The objective of the study was to determine the outcome of surgical resection of metastatic papillary thyroid cancer (PTC) in cervical lymph nodes after failure of initial surgery and I131 therapy.

Design: This was a retrospective clinical study.

Setting: The study was conducted at a university-based tertiary cancer hospital.

Patients: A cohort of 95 consecutive patients with recurrent/persistent PTC in the neck underwent initial reoperation during 1999–2005. All had previous thyroidectomy (±nodal dissection) and I131 therapy. Twenty-five patients with antithyroglobulin (Tg) antibodies were subsequently excluded.

Main Outcome Measures: Biochemical complete remission (BCR) was stringently defined as undetectable TSH-stimulated serum Tg.

Results: A total of 107 lymphadenectomies were undertaken in these 70 patients through January 2010. BCR was initially achieved in 12 patients (17%). Of the 58 patients with detectable postoperative Tg, 28 had a second reoperation and BCR was achieved in five (18%), seven had a third reoperation, and none achieved BCR. No patient achieving BCR had a subsequent recurrence after a mean follow-up of 60 months (range 4–116 months). In addition, two more patients achieved BCR during long-term follow-up without further intervention. In total, 19 patients (27%) achieved BCR and 32 patients (46%) achieved a TSH-stimulated Tg less than 2.0 ng/ml. Patients who did not achieve BCR had significant reduction in Tg after the first (P < 0.001) and second (P = 0.008) operations. No patient developed detectable distant metastases or died from PTC.

Conclusions: Surgical resection of persistent PTC in cervical lymph nodes achieves BCR, when most stringently defined, in 27% of patients, sometimes requiring several surgeries. No biochemical or clinical recurrences occurred during follow-up. In patients who do not achieve BCR, Tg levels were significantly reduced. The long-term durability and impact of this intervention will require further investigation.
Dramaturgy at A.c.t.: An Interview with Dennis Powers and William Ball of the American Conservatory Theatre
Authors: Kleb William
Published: March 2010

Founded by William Ball in 1965, the American Conservatory Theatre settled permanently in San Francisco in 1967; it is now one of the largest and most successful regional repertory companies in the nation, presenting a season of nine classical and modern plays in nightly rotation over thirty-three weeks. A.C.T. has a staff of over 200 with a yearly budget of $4,000,000.00; only 28% of this figure comes from grants and foundations. The position of “dramaturg” at A.C.T. evolved as the pragmatic “literary” needs of the company made themselves apparent over the years. Dennis Powers has been chiefly responsible for this development and during the 1975–1976 season he first officially assumed the title. What follows is an edited record of an interview conducted in February 1978.
The Guthrie: An Interview with Alvin Epstein and Michael Feingold
Authors: Bly Mark , Schechter Joel
Published: March 2010

In 1963, the Tyrone Guthrie Theater was founded in Minneapolis as an alternative to Broadway and its commercialism. Despite occasional financial setbacks, Tyrone Guthrie and his British proteges gradually established it as a major classical repertory theater. Guthrie died in 1971, and the British director Michael Langham succeeded him. Under his artistic direction, the Guthrie Theater stabilized and flourished. When Langham announced he would resign at the end of the 1977 season, an international search for his successor commenced. For the first time during the Guthrie's existence, an American was selected for the post of artistic director. Alvin Epstein, formerly Associate Director of the Yale Repertory Theater, was chosen, beginning a new era in the history of the Guthrie. The repertory for Epstein's first season included such plays as The Pretenders, Hamlet, Tiebele and Her Demon, Bonjour, La, Bonjour, and The Marriage.

The following interview with Alvin Epstein and Literary Manager Michael Feingold was conducted by Joel Schechter and Mark Bly on March 8, 1979 in New York.
Long Wharf: An Interview with Arvin Brown
Authors: Paran Janice , Schechter Joel
Published: March 2010

The Long Wharf Theatre in New Haven, Connecticut opened July 4, 1965, and has since become one of American foremost regional theaters. Its American premiere productions include The Contractor and The Changing Room by David Storey, The National Health and Forget-Me-Not Lane by Peter Nichols, Sizwe Banzi Is Dead and The Island by Athol Fugard, and Spokesong by Stewart Parker. Several Long Wharf productions have been transferred to New York, including Streamers by David Rabe, The Shadow Box by Michael Cristofer, and The Gin Game by D. L. Coburn. Arvin Brown has been Long Wharfs artistic director for eleven years. In that time he has directed regularly: at the Long Wharf; at other regional theaters, and in New York. He has received two Tony Award nominations for his work on Broadway (The National Health and Ah! Wilderness) and has won the Vernon Rice Award for his Off Broadway Production of Long Day's Journey Into Night. Mr. Brown has also worked in television: he directed segments for “Theatre in America” and for the “Visions” series. His most recent directing venture was Strangers, a Broadway play based on the lives of Sinclair Lewis and Dorothy Thompson. The following interview was conducted in March, 1979.
The Rap on Euripides: Creating New Texts in Urban Schools
Authors: Mendelsohn Susan
Published: March 2010

The following interview was conducted after the first stage of an ongoing project, an adaptation of Euripides' Electra incorporating rap music, developed with students at a New Haven high school. Since Susan Mendelsohn has functioned on this project as playwright, dramaturg, teacher, and director, she has taken on the role of interviewer as well, so she could assess the process from every angle and make it complete.
The Speeds of Ambiguity: An Interview with Paul Virilio
Authors: Adams Jason Michael
Published: March 2010

This interview was conducted on the assumption that for all their seeming pessimism, Virilio's technocultural writings are more productively understood as a “Dionysian yes” than as a simple assertion of the no as such. In other words, they presuppose that the specificity of his thought derives not from “the political” itself (as it is explicitly stated) but from the aesthetic as itself political. The interview considers the extent to which Virilio in Deleuze's terms, “has placed the negative at the service of the power of affirming,” even though he has done so in a subterranean manner that largely remains unnoticed by most of his commentators. The assumption that his concept of speed is unidimensional, that he conceives no alternative to the contemporary technoscientific order, and that all forms of mass culture are denounced as equally complicit with late modern power are thus destabilized with the first several questions. Similarly, the notion that Virilio holds that the manipulation of emotion negates democratic possibility as such, that citizenship is entirely incompatible with globalization, and that the only option in our time is to either entirely converge virtual and actual reality or to abandon one in favor of the other are complicated in the remaining ones. Overall, the interview suggests that Virilio himself must be considered as a vector whose concepts are always already in motion, indeed as one who may seem to be making a definitive statement at one moment, but who complicates that very assumption at another. Thus, while the many criticisms of his work are certainly formidable, it seeks to show that they can never adequately represent the composite nature of his thought itself, even when they go to great pains to avoid the suppression of ambiguity.
Implications of pannexin 1 and pannexin 3 for keratinocyte differentiation
Authors: Celetti Steven J. , Cowan Kyle N. , Penuela Silvia , Shao Qing , Churko Jared , Laird Dale W.
Published: March 2010

Pannexin (Panx) 1 and Panx3 are integral membrane proteins that share some sequence homology with the innexin family of invertebrate gap junctions. They are expressed in mammalian skin. Pannexins have been reported to form functional mechanosensitive single-membrane channels, but their importance in regulating cellular function is poorly understood. In this study, Panx1 and Panx3 were detected in the epidermis of 13.5 day embryonic mice. Compared with newborn mice, there was less Panx1 expression in both thin and thick murine skin, whereas Panx3 expression was unchanged. To investigate the role of pannexins in keratinocyte differentiation, we employed rat epidermal keratinocytes (REKs) that have the capacity to differentiate into organotypic epidermis, and engineered them to overexpress Panx1, Panx1-GFP or Panx3. The expression of Panx1 or Panx3 resulted in the increased ability of REKs to take up dye, suggesting that cell-surface channels were formed. Compared with monolayer REKs, endogenous Panx1 levels remained unchanged, whereas the 70 kDa immunoreactive species of Panx3 was greatly increased in the organotypic epidermis. In monolayer cultures, ectopic Panx1 and Panx1-GFP localized to the plasma membrane, whereas Panx3 displayed both intracellular and plasma-membrane profiles. Although both pannexins reduced cell proliferation, only Panx1 disrupted the architecture of the organotypic epidermis and markedly dysregulated cytokeratin 14 expression and localization. Furthermore, ectopic expression of only Panx1 reduced the vital layer thickness of the organotypic epidermis. In summary, Panx1 and Panx3 are coexpressed in the mammalian epidermis, and the regulation of Panx1 plays a key role in keratinocyte differentiation.
Gain of oncogenic function of p53 mutants regulates E-cadherin expression uncoupled from cell invasion in colon cancer cells
Authors: Roger Lauréline , Jullien Laurent , Gire Véronique , Roux Pierre
Published: March 2010

Mutations in the p53 tumour suppressor gene are associated clinically with tumour progression and metastasis. Downregulation of the E-cadherin cell-cell adhesion molecule is a key event for epithelial to mesenchymal transition (EMT) in tumour progression. Here, we show that wild-type p53 induced to adopt a mutant conformation, and hot-spot p53 mutants, which are both transcriptionally inactive, downregulate E-cadherin expression in the colon carcinoma cell line HCT116. Downregulation of E-cadherin occurred concomitantly with the upregulation of Slug and Zeb-1, transcriptional factors known to repress E-cadherin gene expression. In addition, knockdown of Slug and Zeb-1 expression diminished p53-mediated E-cadherin repression. Knocking down endogenous mutant p53 in MDA-MB-231 and SW620 cancer cell lines lacking E-cadherin protein restored the expression of E-cadherin. Complete loss of E-cadherin expression in HCT116 cells induced morphological alterations along with upregulation of vimentin, a mesenchymal marker. These changes characteristic of the EMT phenotype were, however, not sufficient to confer invasiveness in a three-dimensional matrix. Downregulation of E-cadherin by mutant p53 was not required to promote the invasive phenotype induced by inactivation of p53. These findings indicate that independent control of E-cadherin expression and cell motility could be essential molecular events in p53 mutant-induced invasive phenotypes.
CCM3/PDCD10 stabilizes GCKIII proteins to promote Golgi assembly and cell orientation
Authors: Fidalgo Miguel , Fraile María , Pires Ana , Force Thomas , Pombo Celia , Zalvide Juan
Published: March 2010

Mutations in CCM3/PDCD10 result in cerebral cavernous malformations (CCMs), a major cause of cerebral hemorrhage. Despite intense interest in CCMs, very little is known about the function of CCM3. Here, we report that CCM3 is located on the Golgi apparatus, forming a complex with proteins of the germinal center kinase III (GCKIII) family and GM130, a Golgi-resident protein. Cells depleted of CCM3 show a disassembled Golgi apparatus. Furthermore, in wound-healing assays, CCM3-depleted cells cannot reorient the Golgi and centrosome properly, and demonstrate impaired migration. Golgi disassembly after either depletion of CCM3 or dissociation of CCM3 from the GM130-GCKIII complex is the result of destabilization of GCKIII proteins and dephosphorylation of their substrate, 14-3-3ζ. Significantly, the phenotype induced by CCM3 depletion can be reverted by expression of wild-type CCM3, but not by disease-associated mutants. Our findings suggest that Golgi dysfunction and the ensuing abnormalities of cell orientation and migration resulting from CCM3 mutations contribute to CCM pathogenesis.
p53 localizes to intranucleolar regions distinct from the ribosome production compartments
Authors: Krüger Tim , Scheer Ulrich
Published: March 2010

The tumor suppressor p53 has been implicated in the regulation of ribosome biogenesis based on its inhibitory effect on RNA polymerase I (pol I)-dependent transcription. Consistent with this, p53 has been described in nucleoli, albeit under specific experimental conditions. Since data on the intranucleolar localization of p53 are controversial, we have analyzed in detail its subnucleolar distribution. Our results show that p53 does not localize to one of the well-known structural components of the nucleolus involved in ribosome biogenesis, but rather occupies distinct intranucleolar regions that constitute nucleolar cavities. When cells were treated with the proteasome inhibitor MG132, the size and frequency of p53-containing nucleolar cavities increased, and the protein partially colocalized with inactivated proteasomes. Importantly, p53 did not colocalize with pol I at the transcription sites in fibrillar centers (FCs) as has previously been reported. The observed intranucleolar distribution and accumulation of p53 raises the question of how the protein influences rDNA transcription in vivo.
Aberrant expression of a β-catenin gain-of-function mutant induces hyperplastic transformation in the mouse cornea
Authors: Zhang Yujin , Call Mindy K. , Yeh Lung-Kun , Liu Hongshan , Kochel Tyler , Wang I-Jong , Chu Pao-Hsien , Taketo Makoto M. , Jester James V. , Kao Winston W.-Y. , Liu Chia-Yang
Published: March 2010

β-catenin signaling has been shown to play a fundamental role in embryonic development and tumorigenesis. In this study, we investigated the role of β-catenin (Ctnnb1) in corneal homeostasis and tumorigenesis. Conditional expression of a murine Ctnnb1 gain-of-function mutation alone caused corneal neoplasia and neovascularization, resembling human ocular surface squamous neoplasia (OSSN). These corneas displayed an upregulation of cell proliferative markers (PCNA and p63), while presenting downregulation of both the Pax-6 transcription factor and the corneal differentiation marker cytokeratin 12. In addition, the expression of limbal-type keratin 15 ectopically extended to cornea, but the pattern of conjunctival keratin 4 and epidermal keratin 10 were unchanged. Moreover, epithelial E-cadherin and laminins decreased concomitantly with elevated levels of MMP-7. We also noticed a dramatic upregulation of pro-angiogenic factors (Vegf-A, Vegfr1) and angiopoietins in these corneas. Interestingly, all human OSSN specimens examined revealed nuclear β-catenin immunoreactivity. Taken together, these results argue that β-catenin activation is a crucial step during OSSN pathogenesis. Thus, inhibition of β-catenin might be beneficial for treating this disease.
The plant formin AtFH4 interacts with both actin and microtubules, and contains a newly identified microtubule-binding domain
Authors: Deeks Michael J. , Fendrych Matyáš , Smertenko Andrei , Bell Kenneth S. , Oparka Karl , Cvrčková Fatima , Žárský Viktor , Hussey Patrick J.
Published: March 2010

The dynamic behaviour of the actin cytoskeleton in plants relies on the coordinated action of several classes of actin-binding proteins (ABPs). These ABPs include the plant-specific subfamilies of actin-nucleating formin proteins. The model plant species Arabidopsis thaliana has over 20 formin proteins, all of which contain plant-specific regions in place of the GTPase-binding domain, formin homology (FH)3 domain, and DAD and DID motifs found in many fungal and animal formins. We have identified for the first time a plant-specific region of the membrane-integrated formin AtFH4 that mediates an association with the microtubule cytoskeleton. In vitro analysis shows that this region (named the GOE domain) binds directly to microtubules. Overexpressed AtFH4 accumulates at the endoplasmic reticulum membrane and co-aligns the endoplasmic reticulum with microtubules. The FH1 and FH2 domains of formins are conserved in plants, and we show that these domains of AtFH4 nucleate F-actin. Together, these data suggest that the combination of plant-specific and conserved domains enables AtFH4 to function as an interface between membranes and both major cytoskeletal networks.