bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of Human Anti-Murine T-Cell Receptor Antibodies in Both Responding and Nonresponding Patients Enrolled in TCR Gene Therapy Trials
Authors: Davis Jeremy L. , Theoret Marc R. , Zheng Zhili , Lamers Cor H.J. , Rosenberg Steven A. , Morgan Richard A.
Published: December 2010

Purpose: Immune responses to gene-modified cells are a concern in the field of human gene therapy, as they may impede effective treatment. We conducted 2 clinical trials in which cancer patients were treated with lymphocytes genetically engineered to express murine T-cell receptors (mTCR) specific for tumor-associated antigens p53 and gp100.

Experimental Design: Twenty-six patients treated with autologous lymphocytes expressing mTCR had blood and serum samples available for analysis. Patient sera were assayed for the development of a humoral immune response. Adoptive cell transfer characteristics were analyzed to identify correlates to immune response.

Results: Six of 26 (23%) patients' posttreatment sera exhibited specific binding of human anti-mTCR antibodies to lymphocytes transduced with the mTCR. Antibody development was found in both responding and nonresponding patients. The posttreatment sera of 3 of these 6 patients mediated a 60% to 99% inhibition of mTCR activity as measured by a reduction in antigen-specific interferon-γ release. Detailed analysis of posttreatment serum revealed that antibody binding was β-chain specific in 1 patient whereas it was α-chain specific in another.

Conclusions: A subset of patients treated with mTCR-engineered T cells developed antibodies directed to the mTCR variable regions and not to the constant region domains common to all mTCR. Overall, the development of a host immune response was not associated with the level of transduced cell persistence or response to therapy. In summary, patients treated with mTCR can develop an immune response to gene-modified cells in a minority of cases, but this may not affect clinical outcome. Clin Cancer Res; 16(23); 5852–61. ©2010 AACR.
Specific Removal of C-Reactive Protein by Apheresis in a Porcine Cardiac Infarction Model
Authors: Slagman Anna Christine , Bock Christopher , Abdel-Aty Hassan , Vogt Birgit , Gebauer Frank , Janelt Gunnar , Wohlgemuth Franziska , Morgenstern Rene , Yapici Gülcan , Puppe Astrid , Modersohn Diethelm , Mans Dörte , Jerichow Timo , Ott Sascha , Kunze Rudolf , Schrödl Wieland , Janko Christina , Hermann Martin , Kalden Joachim R. , Kern Peter , Parsch Hans , Kirschfink Michael , Schulz-Menger Jeanette , Röttgen Rainer , Unger Juliane K. , Frei Ulrich , Schindler Ralf , Möckel Martin , Sheriff Ahmed
Published: December 2010

Background: C-reactive protein (CRP) is a possible causative factor of the destructive processes observed during the weeks after myocardial infarction. Methods: We developed a clinically relevant animal model including the removal of CRP from blood plasma utilizing a specific CRP adsorber and the visualization of the infarct scar in the living animal by cardiovascular magnetic resonance imaging as a tool to investigate the impact of CRP after acute myocardial infarction. Results: We describe the facets of this model system and kinetics of clinical blood parameters like CRP and troponin. In addition, we demonstrate the potency of CRP apheresis reducing CRP levels by ∼70% in the established treatment system. Conclusion: We showed for the first time that it is possible to conduct apheresis at the following 2 days after acute myocardial infarction in a porcine infarction model and to analyze the infarct by cardiovascular magnetic resonance imaging at day 1 and 14.
Comparison of Interleukin-6 Removal Properties among Hemofilters Consisting of Varying Membrane Materials and Surface Areas: An in vitro Study
Authors: Hirayama Yo , Oda Shigeto , Wakabayashi Kiyohito , Sadahiro Tomohito , Nakamura Masataka , Watanabe Eizo , Tateishi Yoshihisa
Published: December 2010

Background/Aims: We sought to identify the most relevant hemofilter for cytokine removal based on the mechanisms of filtration and adsorption. Methods: Ascites were filtered using four types of hemofilters composed of different membrane materials (polymethyl methacrylate, PMMA, cellulose triacetate, CTA, or polysulfone, PS) and different surface areas (1.0 or 2.1 m2) to investigate the rate of interleukin-6 (IL-6) filtration. Next, ascites were perfused through each hemofilter without obtaining a filtrate to study each filter’s adsorptive capability. Results: The PMMA hemofilters resulted in a marginal observed IL-6 filtration rates, whereas the CTA and PS hemofilters resulted in highly effective IL-6 filtration. Regarding the IL-6 adsorptive capabilities of the filters, the PMMA hemofilter with a large surface area showed the highest level of IL-6 clearance. Conclusion: The present findings suggest that when cytokine removal based on filtration is desired, CTA or PS hemofilters should be selected. When IL-6 removal based on adsorption is desired, a PMMA hemofilter with a large surface area should be selected.
Shear-dependent suppression of platelet thrombus formation by phosphodiesterase 3 inhibition requires low levels of concomitant Gs-coupled receptor stimulation
Authors: Yoshida Hideo , Okamura Yosuke , Watanabe Naohide , Ikeda Yasuo , Handa Makoto
Published: December 2010

Phosphodiesterase (PDE)3 inhibitors exert potent antiplatelet effects through maintaining elevated intracellular cyclic adenosine monophosphate levels, but do not prolong bleeding time. To resolve this discrepancy, we hypothesised that PDE3 inhibitors effectively suppress shear-induced platelet thrombus formation initiated by the interaction of the platelet receptor GPIb/V/IX with its ligand, von Willebrand factor (VWF), since arterial thrombosis is more dependent on shear stress as compared with haemostatic plug formation. To test the hypothesis, we compared the in vitro effects of K-134 (a PDE3 inhibitor), tirofiban (a GPIIb/IIIa inhibitor) and acetylsalicylic acid (ASA) on ristocetin-induced platelet aggregation and platelet thrombus formation on VWF or collagen surfaces under flow conditions. K-134 inhibited GPIIb/IIIa-dependent platelet aggregation to the same extent as tirofiban and more potently than ASA. Likewise, K-134 and tirofiban effectively inhibited stable platelet thrombus formation (platelet firm adhesion and subsequent aggregation) on the VWF or collagen surface under high shear, but ASA only inhibited aggregation. Notably, inhibition by K-134 became evident only when a low concentration of PGE1 was present. These inhibitors did not block shear-induced initial platelet contact with VWF via GPIb/V/IX. In contrast, under low shear, the inhibitory effects of K-134 on platelet aggregation on the collagen surface were lower than tirofiban or ASA. The observed shear-dependent suppression of platelet thrombus formation by PDE3 inhibitor in the presence of low levels of adenylate cyclase stimulator may contribute to high therapeutic benefit with low risk of bleeding.
Outcomes of Percutaneous Endoscopic Gastrostomy Tube Placement Using a T‐Fastener Gastropexy Device in Head and Neck and Esophageal Cancer Patients
Authors: Chadha Krishdeep Singh , Thatikonda Chandana , Schiff Michael , Nava Hector , Sitrin Michael D.
Published: December 2010

Background: Percutaneous endoscopic gastrostomy (PEG) tube placement by the pull technique in head, neck, and esophageal cancer patients has a high complication rate, particularly in infections, and a small risk of tumor implantation. The T‐fastener gastropexy technique uses a transabdominal approach to place the PEG device. Objectives: The objective of this study was to review the clinical outcomes and complications related to endoscopic PEG placement with the T‐fastener gastropexy technique in patients with head, neck, and esophageal cancers. Methods: This study was a retrospective review of all patients with head, neck, and esophageal cancers with PEG placement from January 1998 to June 2008. Clinical data including patient's age, gender, type and stage of cancer, date of PEG placement and removal, and reason for PEG removal was recorded. Results: The study group consisted of a total of 356 patients of which 244 were male and 112 were female with a mean age of 63.3 years. There were 276 patients with head and neck cancer, 75 patients with esophageal cancer, and 5 with gastro‐esophageal junction cancer. Staging data was available for 326 patients of which 56 (17.1%) had early stage disease (stage 1 and 2) and 270 patients (82.9%) had late stage (3 and 4) disease. None of these patients received antibiotic prophylaxis prior to PEG placement. Cellulitis around the PEG site occurred in 8.4% of cases and an abscess in 3.7% of cases. Only 4 (1.2%) infectious complications were within the first 30 days of PEG placement, and there were no intraoperative deaths or cases of tumor implantation. Conclusions: In the authors’ experience, the T‐fastener gastropexy technique for PEG placement in head, neck, and esophageal cancer patients carried a low overall complication rate and compared favorably with the results of pull method reported in the literature.
Compensatory Increased Enteral Feeding Goal Rates
Authors: Lichtenberg Kristen , Guay‐Berry Patricia , Pipitone Angela , Bondy Anna , Rotello Leo
Published: December 2010

Background: Traditionally, enteral nutrition (EN) goal rates have been calculated based on an intended continuous 24‐hour infusion rate. Many factors in the care of critically ill patients result in interruption of EN infusions, often for several hours daily, which may lead to significant underfeeding. The objective of this study was to evaluate the difference of daily EN volume deficits between a traditionally calculated infusion rate and a compensatory, higher calculated infusion rate in which the 24‐hour volume was delivered over a 20‐hour infusion period. Methods: Data collection consisted of daily EN volume deficit (intended volume – actual volume infused), based on intensive care unit nursing flow sheets. The primary outcome was daily EN volume deficit from a standard 24‐hour calculated goal rate, compared with volume deficit from delivery of the same volume over 20 hours. For the 20‐hour group, the calculated daily requirement of EN was divided by 20 rather than 24 for the higher hourly rate but still delivered for 24 hours. Results: One hundred ten baseline (24‐hour) EN days were evaluated with a mean infusion volume deficit of (±)247 mL/d. This compared with 158 patients in the 20‐hour infusion group, in which the mean volume deficit was (±)45 mL/d (P < .001). Enteral nutrition was most often held for extubation or procedures. A higher level of overfeeding was noted in the 20‐hour infusion group. Conclusion: Calculating and prescribing higher EN infusion rates, assuming 20 hours of actual infusion daily, promoted delivery of optimal nutrient provisions and avoidance of unintended malnutrition by significantly reducing caloric deficit from frequent EN holding.
Clinical Relevancy of the Levothyroxine–Continuous Enteral Nutrition Interaction
Authors: Dickerson Roland N. , Maish George O. , Minard Gayle , Brown Rex O.
Published: December 2010

Background: The objective of this study was to determine if patients requiring levothyroxine therapy develop hypothyroidism during concurrent continuous enteral nutrition (EN). Methods: Adult patients with a history of hypothyroidism, given levothyroxine via the feeding tube at the same dose given prior to hospital admission and who were referred to the nutrition support service for EN were evaluated. Thyroid function tests (TFTs) were performed prior to administration of levothyroxine–continuous EN, then approximately weekly thereafter. Patients who received less than 14 days of concurrent EN‐levothyroxine therapy were excluded from the analysis. Results: Thirteen patients who received 20 ± 5 days of concurrent EN and levothyroxine were evaluated. Two patients developed subclinical hypothyroidism (thyrotropin [TSH] >6 and <10 mcIU/mL + normal fT4), and 6 developed overt hypothyroidism (TSH >10 mcIU/mL + low fT4) within 2 to 3 weeks of concurrent EN‐levothyroxine therapy. Five patients remained euthyrotic. Differences between those who developed subclinical or overt hypothyroidism versus those who remained euthyrotic could not be explained by age, weight, levothyroxine dose, type of EN formula, or amount of EN received. Conclusions: More than half of the patients receiving concurrent levothyroxine–continuous EN developed subclinical or overt hypothyroidism requiring therapeutic intervention. Routine weekly monitoring of TFTs for patients receiving concurrent levothyroxine–continuous EN is recommended.
Article Commentary: Conformity to Traditional Mediterranean Diet and Breast Cancer Risk in the Greek EPIC (European Prospective Investigation into Cancer and nutrition) Cohort
Authors: Mullin Gerard E.
Published: December 2010

Conformity to Traditional Mediterranean Diet and Breast Cancer Risk in the Greek EPIC (European Prospective Investigation into Cancer and nutrition) Cohort Trichopoulou A Bamia C Lagiou P Trichopoulos D Am J Clin Nutr. doi: 10. 3945/ajcn.2010.29619.
The Standard American Diet and Its Relationship to the Health Status of Americans
Authors: Grotto David , Zied Elisa
Published: December 2010

The Standard American Diet (SAD) has long been implicated in contributing to the health challenges experienced in the United States. Significant changes to the SAD have occurred since the 1950s, including a greater abundance and accessibility to calorie‐dense and nutrient‐poor food and beverage choices. The disparity of present consumption patterns to diet and nutrition recommendations from the Dietary Guidelines for Americans are addressed.
A garter snake transcriptome: pyrosequencing, de novo assembly, and sex-specific differences
Authors: Schwartz Tonia S , Tae Hongseok , Yang Youngik , Mockaitis Keithanne , Van Hemert John L , Proulx Stephen R , Choi Jeong-Hyeon , Bronikowski Anne M
Published: December 2010

Background

The reptiles, characterized by both diversity and unique evolutionary adaptations, provide a comprehensive system for comparative studies of metabolism, physiology, and development. However, molecular resources for ectothermic reptiles are severely limited, hampering our ability to study the genetic basis for many evolutionarily important traits such as metabolic plasticity, extreme longevity, limblessness, venom, and freeze tolerance. Here we use massively parallel sequencing (454 GS-FLX Titanium) to generate a transcriptome of the western terrestrial garter snake (Thamnophis elegans) with two goals in mind. First, we develop a molecular resource for an ectothermic reptile; and second, we use these sex-specific transcriptomes to identify differences in the presence of expressed transcripts and potential genes of evolutionary interest.

Results

Using sex-specific pools of RNA (one pool for females, one pool for males) representing 7 tissue types and 35 diverse individuals, we produced 1.24 million sequence reads, which averaged 366 bp in length after cleaning. Assembly of the cleaned reads from both sexes with NEWBLER and MIRA resulted in 96,379 contigs containing 87% of the cleaned reads. Over 34% of these contigs and 13% of the singletons were annotated based on homology to previously identified proteins. From these homology assignments, additional clustering, and ORF predictions, we estimate that this transcriptome contains ~13,000 unique genes that were previously identified in other species and over 66,000 transcripts from unidentified protein-coding genes. Furthermore, we use a graph-clustering method to identify contigs linked by NEWBLER-split reads that represent divergent alleles, gene duplications, and alternatively spliced transcripts. Beyond gene identification, we identified 95,295 SNPs and 31,651 INDELs. From these sex-specific transcriptomes, we identified 190 genes that were only present in the mRNA sequenced from one of the sexes (84 female-specific, 106 male-specific), and many highly variable genes of evolutionary interest.

Conclusions

This is the first large-scale, multi-organ transcriptome for an ectothermic reptile. This resource provides the most comprehensive set of EST sequences available for an individual ectothermic reptile species, increasing the number of snake ESTs 50-fold. We have identified genes that appear to be under evolutionary selection and those that are sex-specific. This resource will assist studies on gene expression and comparative genomics, and will facilitate the study of evolutionarily important traits at the molecular level.
Elevated AKR1C3 expression promotes prostate cancer cell survival and prostate cell-mediated endothelial cell tube formation: implications for prostate cancer progressioan
Authors: Dozmorov Mikhail G , Azzarello Joseph T , Wren Jonathan D , Fung Kar-Ming , Yang Qing , Davis Jeffrey S , Hurst Robert E , Culkin Daniel J , Penning Trevor M , Lin Hsueh-Kung
Published: December 2010

Background

Aldo-keto reductase (AKR) 1C family member 3 (AKR1C3), one of four identified human AKR1C enzymes, catalyzes steroid, prostaglandin, and xenobiotic metabolism. In the prostate, AKR1C3 is up-regulated in localized and advanced prostate adenocarcinoma, and is associated with prostate cancer (PCa) aggressiveness. Here we propose a novel pathological function of AKR1C3 in tumor angiogenesis and its potential role in promoting PCa progression.

Methods

To recapitulate elevated AKR1C3 expression in cancerous prostate, the human PCa PC-3 cell line was stably transfected with an AKR1C3 expression construct to establish PC3-AKR1C3 transfectants. Microarray and bioinformatics analysis were performed to identify AKR1C3-mediated pathways of activation and their potential biological consequences in PC-3 cells. Western blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and anin vitroMatrigel angiogenesis assays were applied to validate the pro-angiogenic activity of PC3-AKR1C3 transfectants identified by bioinformatics analysis.

Results

Microarray and bioinformatics analysis suggested that overexpression of AKR1C3 in PC-3 cells modulates estrogen and androgen metabolism, activates insulin-like growth factor (IGF)-1 and Akt signaling pathways, as well as promotes tumor angiogenesis and aggressiveness. Levels of IGF-1 receptor (IGF-1R) and Akt activation as well as vascular endothelial growth factor (VEGF) expression and secretion were significantly elevated in PC3-AKR1C3 transfectants in comparison to PC3-mock transfectants. PC3-AKR1C3 transfectants also promoted endothelial cell (EC) tube formation on Matrigel as compared to the AKR1C3-negative parental PC-3 cells and PC3-mock transfectants. Pre-treatment of PC3-AKR1C3 transfectants with a selective IGF-1R kinase inhibitor (AG1024) or a non-selective phosphoinositide 3-kinases (PI3K) inhibitor (LY294002) abolished ability of the cells to promote EC tube formation.

Conclusions

Bioinformatics analysis followed by functional genomics demonstrated that AKR1C3 overexpression promotes angiogenesis and aggressiveness of PC-3 cells. These results also suggest that AKR1C3-mediated tumor angiogenesis is regulated by estrogen and androgen metabolism with subsequent IGF-1R and Akt activation followed by VEGF expression in PCa cells.
Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya
Authors: Gatei Wangeci , Kariuki Simon , Hawley William , ter Kuile Feiko , Terlouw Dianne , Phillips-Howard Penelope , Nahlen Bernard , Gimnig John , Lindblade Kim , Walker Edward , Hamel Mary , Crawford Sara , Williamson John , Slutsker Laurence , Shi Ya Ping
Published: December 2010

Background

Insecticide-treated bed nets (ITNs) reduce malaria transmission and are an important prevention tool. However, there are still information gaps on how the reduction in malaria transmission by ITNs affects parasite genetics population structure. This study examined the relationship between transmission reduction from ITN use and the population genetic diversity of Plasmodium falciparum in an area of high ITN coverage in western Kenya.

Methods

Parasite genetic diversity was assessed by scoring eight single copy neutral multilocus microsatellite (MS) markers in samples collected from P. falciparum- infected children (< five years) before introduction of ITNs (1996, baseline, n = 69) and five years after intervention (2001, follow-up, n = 74).

Results

There were no significant changes in overall high mixed infections and unbiased expected heterozygosity between baseline (%MA = 94% and He = 0.75) and follow up (%MA = 95% and He = 0.79) years. However, locus specific analysis detected significant differences for some individual loci between the two time points. Pfg377 loci, a gametocyte-specific MS marker showed significant increase in mixed infections and He in the follow up survey (%MA = 53% and He = 0.57) compared to the baseline (%MA = 30% and He = 0.29). An opposite trend was observed in the erythrocyte binding protein (EBP) MS marker. There was moderate genetic differentiation at the Pfg377 and TAA60 loci (FST = 0.117 and 0.137 respectively) between the baseline and post-ITN parasite populations. Further analysis revealed linkage disequilibrium (LD) of the microsatellites in the baseline (14 significant pair-wise tests and I

S

A
= 0.016) that was broken in the follow up parasite population (6 significant pairs and I

S

A
= 0.0003). The locus specific change in He, the moderate population differentiation and break in LD between the baseline and follow up years suggest an underlying change in population sub-structure despite the stability in the overall genetic diversity and multiple infection levels.

Conclusions

The results from this study suggest that although P. falciparum population maintained an overall stability in genetic diversity after five years of high ITN coverage, there was significant locus specific change associated with gametocytes, marking these for further investigation.
Rac1-mediated signaling plays a central role in secretion-dependent platelet aggregation in human blood stimulated by atherosclerotic plaque
Authors: Dwivedi Suman , Pandey Dharmendra , Khandoga Anna L , Brandl Richard , Siess Wolfgang
Published: December 2010

Background

Platelet activation requires rapid remodeling of the actin cytoskeleton which is regulated by small GTP-binding proteins. By using the Rac1-specific inhibitor NSC23766, we have recently found that Rac1 is a central component of a signaling pathway that regulates dephosphorylation and activation of the actin-dynamising protein cofilin, dense and α-granule secretion, and subsequent aggregation of thrombin-stimulated washed platelets.

Objectives

To study whether NSC23766 inhibits stimulus-induced platelet secretion and aggregation in blood.

Methods

Human platelet aggregation and ATP-secretion were measured in hirudin-anticoagulated blood and platelet-rich plasma (PRP) by using multiple electrode aggregometry and the Lumi-aggregometer. Platelet P-selectin expression was quantified by flow cytometry.

Results

NSC23766 (300 μM) inhibited TRAP-, collagen-, atherosclerotic plaque-, and ADP-induced platelet aggregation in blood by 95.1%, 93.4%, 92.6%, and 70%, respectively. The IC50 values for inhibition of TRAP-, collagen-, and atherosclerotic plaque-, were 50 ± 18 μM, 64 ± 35 μM, and 50 ± 30 μM NSC23766 (mean ± SD, n = 3-7), respectively. In blood containing RGDS to block integrin αIIbβ3-mediated platelet aggregation, NSC23766 (300 μM) completely inhibited P-selectin expression and reduced ATP-secretion after TRAP and collagen stimulation by 73% and 85%, respectively. In ADP-stimulated PRP, NSC23766 almost completely inhibited P-selectin expression, in contrast to aspirin, which was ineffective. Moreover, NSC23766 (300 μM) decreased plaque-stimulated platelet adhesion/aggregate formation under arterial flow conditions (1500s-1) by 72%.

Conclusions

Rac1-mediated signaling plays a central role in secretion-dependent platelet aggregation in blood stimulated by a wide array of platelet agonists including atherosclerotic plaque. By specifically inhibiting platelet secretion, the pharmacological targeting of Rac1 could be an interesting approach in the development of future antiplatelet drugs.
An integrated assessment of wild vegetable resources in Inner Mongolian Autonomous Region, China
Authors: Wujisguleng Wujisguleng , Khasbagen Khasbagen
Published: December 2010

Background

This paper was based on ethnobotanical investigations conducted from 2004-2006 in Inner Mongolian Autonomous Region of northern China. Today, due to their nutritious and relatively pollution-free characteristics, wild vegetables are playing an increasingly important role in peoples' health and well-being. This paper aims to provide scientific clues for the selection of special and high quality wild vegetables species.

Methods

An ethnobotanical study, consisting of a literature survey, open-ended and semi-structured interviews, and collection and identification of voucher specimens was carried out to gather information on wild vegetables in Inner Mongolia. Next, an integrated assessment of 90 species of wild vegetables was performed using the linearity weighted integrative mathematical analysis method.

Results

According to an integrated assessment of 90 species of wild vegetables in Inner Mongolia, there are 5 species with the highest integrated value, 40 species of high-integrated value, 43 species of general integrated value, and 2 species of low value. The results indicate that the vast majority of wild vegetables have high value in Inner Mongolia.

Conclusions

Inner Mongolia is rich in wild vegetable resources. A comprehensive assessment indicates that the vast majority of wild vegetables are of high value. However, these wild vegetables are seldom collected or cultivated by local people. Most of the collected species require further research and investigation into their nutrient content, toxicity and ethnobotany to illuminate their potential as new cultivars or products.
Prediction of the responsiveness to pharmacological chaperones: lysosomal human alpha-galactosidase, a case of study
Authors: Andreotti Giuseppina , Guarracino Mario R , Cammisa Marco , Correra Antonella , Cubellis Maria Vittoria
Published: December 2010

Background

The pharmacological chaperones therapy is a promising approach to cure genetic diseases. It relies on substrate competitors used at sub-inhibitory concentration which can be administered orally, reach difficult tissues and have low cost. Clinical trials are currently carried out for Fabry disease, a lysosomal storage disorder caused by inherited genetic mutations of alpha-galactosidase. Regrettably, not all genotypes respond to these drugs.

Results

We collected the experimental data available in literature on the enzymatic activity of ninety-six missense mutants of lysosomal alpha-galactosidase measured in the presence of pharmacological chaperones. We associated with each mutation seven features derived from the analysis of 3D-structure of the enzyme, two features associated with their thermo-dynamic stability and four features derived from sequence alone. Structural and thermodynamic analysis explains why some mutants of human lysosomal alpha-galactosidase cannot be rescued by pharmacological chaperones: approximately forty per cent of the non responsive cases examined can be correctly associated with a negative prognostic feature. They include mutations occurring in the active site pocket, mutations preventing disulphide bridge formation and severely destabilising mutations. Despite this finding, prediction of mutations responsive to pharmacological chaperones cannot be achieved with high accuracy relying on combinations of structure- and thermodynamic-derived features even with the aid of classical and state of the art statistical learning methods.

We developed a procedure to predict responsive mutations with an accuracy as high as 87%: the method scores the mutations by using a suitable position-specific substitution matrix. Our approach is of general applicability since it does not require the knowledge of 3D-structure but relies only on the sequence.

Conclusions

Responsiveness to pharmacological chaperones depends on the structural/functional features of the disease-associated protein, whose complex interplay is best reflected on sequence conservation by evolutionary pressure. We propose a predictive method which can be applied to screen novel mutations of alpha galactosidase. The same approach can be extended on a genomic scale to find candidates for therapy with pharmacological chaperones among proteins with unknown tertiary structures.
Autocrine transforming growth factor β signaling regulates extracellular signal-regulated kinase 1/2 phosphorylation via modulation of protein phosphatase 2A expression in scleroderma fibroblasts
Authors: Samuel Glady H , Bujor Andreea M , Nakerakanti Sashidhar S , Hant Faye N , Trojanowska Maria
Published: December 2010

Background

During scleroderma (SSc) pathogenesis, fibroblasts acquire an activated phenotype characterized by enhanced production of extracellular matrix (ECM) and constitutive activation of several major signaling pathways including extracellular signal-related kinase (ERK1/2). Several studies have addressed the role of ERK1/2 in SSc fibrosis however the mechanism of its prolonged activation in SSc fibroblasts is still unknown. Protein phosphatase 2A (PP2A) is a key serine threonine phosphatase responsible for dephosphorylation of a wide array of signaling molecules. Recently published microarray data from cultured SSc fibroblasts suggests that the catalytic subunit (C-subunit) of PP2A is downregulated in SSc. In this study we examined the role and regulation of PP2A in SSc fibroblasts in the context of ERK1/2 phosphorylation and matrix production.

Results

We show for the first time that PP2A mRNA and protein expression are significantly reduced in SSc fibroblasts and correlate with an increase in ERK1/2 phosphorylation and collagen expression. Furthermore, transforming growth factor β (TGFβ), a major profibrotic cytokine implicated in SSc fibrosis, downregulates PP2A expression in healthy fibroblasts. PP2A-specific small interfering RNA (siRNA) was utilized to confirm the role of PP2A in ERK1/2 dephosphorylation in dermal fibroblasts. Accordingly, blockade of autocrine TGFβ signaling in SSc fibroblasts using soluble recombinant TGFβ receptor II (SRII) restored PP2A levels and decreased ERK1/2 phosphorylation and collagen expression. In addition, we observed that inhibition of ERK1/2 in SSc fibroblasts increased PP2A expression suggesting that ERK1/2 phosphorylation also contributes to maintaining low levels of PP2A, leading to an even further amplification of ERK1/2 phosphorylation.

Conclusions

Taken together, these studies suggest that decreased PP2A levels in SSc is a result of constitutively activated autocrine TGFβ signaling and could contribute to enhanced phosphorylation of ERK1/2 and matrix production in SSc fibroblasts.
Global turnover of histone post-translational modifications and variants in human cells
Authors: Zee Barry M , Levin Rebecca S , DiMaggio Peter A , Garcia Benjamin A
Published: December 2010

Background

Post-translational modifications (PTMs) on the N-terminal tails of histones and histone variants regulate distinct transcriptional states and nuclear events. Whereas the functional effects of specific PTMs are the current subject of intense investigation, most studies characterize histone PTMs/variants in a non-temporal fashion and very few studies have reported kinetic information about these histone forms. Previous studies have used radiolabeling, fluorescence microscopy and chromatin immunoprecipitation to determine rates of histone turnover, and have found interesting correlations between increased turnover and increased gene expression. Therefore, histone turnover is an understudied yet potentially important parameter that may contribute to epigenetic regulation. Understanding turnover in the context of histone modifications and sequence variants could provide valuable additional insight into the function of histone replacement.

Results

In this study, we measured the metabolic rate of labeled isotope incorporation into the histone proteins of HeLa cells by combining stable isotope labeling of amino acids in cell culture (SILAC) pulse experiments with quantitative mass spectrometry-based proteomics. In general, we found that most core histones have similar turnover rates, with the exception of the H2A variants, which exhibit a wider range of rates, potentially consistent with their epigenetic function. In addition, acetylated histones have a significantly faster turnover compared with general histone protein and methylated histones, although these rates vary considerably, depending on the site and overall degree of methylation. Histones containing transcriptionally active marks have been consistently found to have faster turnover rates than histones containing silent marks. Interestingly, the presence of both active and silent marks on the same peptide resulted in a slower turnover rate than either mark alone on that same peptide. Lastly, we observed little difference in the turnover between nearly all modified forms of the H3.1, H3.2 and H3.3 variants, with the notable exception that H3.2K36me2 has a faster turnover than this mark on the other H3 variants.

Conclusions

Quantitative proteomics provides complementary insight to previous work aimed at quantitatively measuring histone turnover, and our results suggest that turnover rates are dependent upon site-specific post-translational modifications and sequence variants.
Experiencing antiretroviral adherence: helping healthcare staff better understand adherence to paediatric antiretrovirals
Authors: Phelps Benjamin R , Hathcock Sarah J , Werdenberg Jennifer , Schutze Gordon E
Published: December 2010

Background

Lack of adherence to antiretroviral medications is one of the key challenges for paediatric HIV care and treatment programmes. There are few hands‐on opportunities for healthcare workers to gain awareness of the psychosocial and logistic challenges that caregivers face when administering daily antiretroviral therapy to children. This article describes an educational activity that allows healthcare workers to simulate this caregiver role.

Methods

Paediatric formulations of several antiretroviral medications were dispensed to a convenience sample of staff at the Baylor College of Medicine‐Bristol‐Myers Squibb Children's Clinical Center of Excellence in Mbabane, Swaziland. The amounts of the medications remaining were collected and measured one week later. Adherence rates were calculated. Following the exercise, a brief questionnaire was administered to all staff participants.

Results

The 27 clinic staff involved in the exercise had varying and low adherence rates over the week during which the exercise was conducted. Leading perceived barriers to adherence included: “family friends don't help me remember/tell me I shouldn't take it” and “forgot”. Participants reported that the exercise was useful as it allowed them to better address the challenges faced by paediatric patients and caregivers.

Conclusions

Promoting good adherence practices among caregivers of children on antiretrovirals is challenging but essential in the treatment of paediatric HIV. Participants in this exercise achieved poor adherence rates, but identified with many of the barriers commonly reported by caregivers. Simulations such as this have the potential to promote awareness of paediatric ARV adherence issues among healthcare staff and ultimately improve adherence support and patient outcomes.
The importance of clinic attendance in the first six months on antiretroviral treatment: a retrospective analysis at a large public sector HIV clinic in South Africa
Authors: Brennan Alana T , Maskew Mhairi , Sanne Ian , Fox Matthew P
Published: December 2010

Background

Adherence to care and treatment are essential for HIV‐infected individuals to benefit from antiretroviral therapy (ART). We sought to quantify the effects on treatment outcomes of missing visits soon after initiating ART.

Methods

We analyzed data from HIV‐infected patients initiating ART at Themba Lethu Clinic, Johannesburg, South Africa, from April 2004 to August 2008. We used log‐binomial regression to evaluate the relative risk of missing visits during the first six months of ART on immunological response and virologic suppression. Cox models were used to evaluate the relationship between missed visits and mortality and loss to follow up over 12 months.

Results

Of 4476 patients, 65% missed no visits, while 26% missed one visit, 7% missed two and 1.6% missed three or more visits during the first six months on treatment. Patients who missed three or more medical or antiretroviral (ARV) visits had a two‐fold increased risk of poor CD4 response by six months, while the risk of failing to achieve virologic suppression by six months increased two‐ to five‐fold among patients who missed two and three or more medical or ARV visits. Adjusted Cox models showed that patients who missed two (HR 2.1; 95% CI: 1.0‐4.3) and three or more (HR 4.7; 95% CI: 1.4‐16.2) medical visits had an increased risk of death, while those who missed two ARV (HR 3.8; 95% CI: 2.5‐5.8) or three or more medical (HR 3.0; 95% CI: 1.1‐8.1) visits had an increased risk of loss to follow up.

Conclusions

Thirty‐five percent of patients missed one or more visits in the first six months on treatment, increasing their risk of poorer outcomes. These patients could be targeted for additional adherence counselling to help improve ART outcomes.
Impact of Age at Diagnosis on Prostate Cancer Treatment and Survival
Authors: Bechis Seth K. , Carroll Peter R. , Cooperberg Matthew R.
Published: December 2010

Purpose

Older men are more likely to be diagnosed with high-risk prostate cancer and to have lower overall survival. As a result, age often plays a role in treatment choice. However, the relationships among age, disease risk, and prostate cancer–specific survival have not been well established.

Patients and Methods

We studied men in the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) database with complete risk, treatment, and follow-up information. High-risk patients were identified by using the validated Cancer of the Prostate Risk Assessment (CAPRA) score. Competing risks regression was used to identify the independent impact of age on cancer-specific survival. We also analyzed the effect of local treatment on survival among older men with high-risk disease.

Results

In all, 26% of men age ≥ 75 years presented with high-risk disease (CAPRA score 6 to 10). Treatment varied markedly with age across risk strata; older men were more likely to receive androgen deprivation monotherapy. Controlling for treatment modality alone, or for treatment and risk, age did not independently predict cancer-specific survival. Furthermore, controlling for age, comorbidity, and risk, older men with high-risk tumors receiving local therapy had a 46% reduction in mortality compared with those treated conservatively.

Conclusion

Older patients are more likely to have high-risk prostate cancer at diagnosis and less likely to receive local therapy. Indeed, underuse of potentially curative local therapy among older men with high-risk disease may in part explain observed differences in cancer-specific survival across age strata. These findings support making decisions regarding treatment on the basis of disease risk and life expectancy rather than on chronologic age.
Our Tangled Web
Authors: Nardo Anna K.
Published: December 2010

In research-intensive universities, a complex web of inter-relations between mandates for research productivity and for general education teaching perpetuates the division into a two-tiered faculty described in the ADE survey of staffing patterns in departments of English. Other published and planned MLA and ADE reports—specifically, on the evaluation of scholarship for tenure and promotion, and on the master's degree—further illuminate the inter-relations between graduate education and general education staffing practices. MLA (in its “Academic Workforce Advocacy Kit”) and the Coalition for the Academic Workforce (in its issue brief entitled “On Faculty Serving All Students”) provide leadership for productive workforce changes.
Teaching Native Autobiographies as Acts of Narrative Resistance
Authors: Beard Laura J.
Published: December 2010

“Teaching Native Autobiographies as Acts of Narrative Resistance” is written for non-specialists in Native literature who include a Native-authored work in their classes. This article offers strategies to increase our understanding and appreciation of Native literature by opening up classroom discussions to critical issues in the study of Native literary texts.
Engaging Death, Drama, the Classroom, and Real Life
Authors: Curtin Adrian
Published: December 2010

In teaching a course on death in modern theater to fifteen undergraduates, I had to engage with a real-life death “drama” (the death of a peer of my students) that impinged on my class, presenting me with an uncomfortable pedagogical conundrum. I had to re-think my objectives as an instructor and my conception of the classroom as a safe space. In this article, I rehearse this complicated and potentially fractious class scenario and scrutinize my approach to it. I investigate the potential merits of thinking, feeling, and working through crisis in a classroom situation, thereby fashioning a type of pedagogical “third space” in which ideational and circumstantial crossover is allowed. Some of the issues that arise are the ways in which we can situate pedagogy in praxis with “real life” and what challenges are provided?
The interactive presentation of 3D information obtained from reconstructed datasets and 3D placement of single histological sections with the 3D portable document format
Authors: de Boer Bouke A. , Soufan Alexandre T. , Hagoort Jaco , Mohun Timothy J. , van den Hoff Maurice J. B. , Hasman Arie , Voorbraak Frans P. J. M. , Moorman Antoon F. M. , Ruijter Jan M.
Published: December 2010

Interpretation of the results of anatomical and embryological studies relies heavily on proper visualization of complex morphogenetic processes and patterns of gene expression in a three-dimensional (3D) context. However, reconstruction of complete 3D datasets is time consuming and often researchers study only a few sections. To help in understanding the resulting 2D data we developed a program (TRACTS) that places such arbitrary histological sections into a high-resolution 3D model of the developing heart. The program places sections correctly, robustly and as precisely as the best of the fits achieved by five morphology experts. Dissemination of 3D data is severely hampered by the 2D medium of print publication. Many insights gained from studying the 3D object are very hard to convey using 2D images and are consequently lost or cannot be verified independently. It is possible to embed 3D objects into a pdf document, which is a format widely used for the distribution of scientific papers. Using the freeware program Adobe Reader to interact with these 3D objects is reasonably straightforward; creating such objects is not. We have developed a protocol that describes, step by step, how 3D objects can be embedded into a pdf document. Both the use of TRACTS and the inclusion of 3D objects in pdf documents can help in the interpretation of 2D and 3D data, and will thus optimize communication on morphological issues in developmental biology.
Lewtonian space: Val Lewton's films and the new space of horror
Authors: Telotte J. P
Published: December 2010

During the 1940s the Val Lewton unit at RKO Studios produced a string of horror films that were highly lauded for their subtle approach to the genre, which represented a distinct break from earlier horror films that were characterized by their emphasis on monstrous figures and exaggerated, expressionist-influenced imagery. A significant element of that influence, however, has so far gone unexplored, particularly what we might term thespaceof horror. Drawing on architectural developments and theory in the late modernist period, particularly as articulated by Anthony Vidler, this article examines how the Lewton films drew on this new sense of space, a space that emphasized not structures or containment, but rather the emerging psychological and social dimensions of the era. Because of wartime restrictions and the economical practices of B-film production, the Lewton films (and as illustrations this article draws examples from each of the three directors who worked in this unit Jacques Tourneur, Mark Robson and Robert Wise) almost had to function in a different register than the earlier wave of horror films with their emphases on old dark houses and castles, on werewolves and vampires. The new spatial strategy that they evolved, however, not only accommodated those period and industrial limitations, but also opened up a new possibility for representing narratives of power and dread one that mobilized space as a placeholder for all of our psychic projections and fears.
Lost in translation? Ghost music in recent Japanese Kaidan films and their Hollywood remakes
Authors: Wierzbicki James
Published: December 2010

Western audiences in recent years have been intrigued by the cinematic phenomenon popularly known as J-Horror. Critical attention has been paid to more to the Hollywood remakes than to the original Japanese films. Comparing such Hollywood films as The Ring, The Grudge, and Dark
Water with their models, this essay focuses on the various soundtracks' use of a particular ghostly noise. It argues that whereas the soundtracks of the Japanese Kaidan films in various ways sustain an aesthetic/theatrical tradition that is centuries old, the Hollywood remakes miss
an important point by appropriating the sound but not its context.
Time is wasting: con/sequence and s/pace in the Saw series
Authors: Jones Steve
Published: December 2010

Horror film sequels have not received as much serious critical attention as they deserve this is especially true of the Saw franchise, which has suffered a general dismissal under the derogatory banner Torture Porn. In this article I use detailed textual analysis of the Saw
series to expound how film sequels employ and complicate expected temporal and spatial relations in particular, I investigate how the Saw sequels tie space and time into their narrative, methodological and moral sensibilities. Far from being a gimmick or a means of ensuring loyalty
to the franchise (one has to be familiar with the events of previous episodes to ascertain what is happening), it is my contention that the Saw cycle directly requests that we examine the nature of space and time, in terms of both cinematic technique and our lived, off-screen temporal/spatial
orientations.
Make a copy, pass it on: The Ring Two and the Ghost of Verbinski
Authors: Hu Brian
Published: December 2010

As a Hollywood production helmed by Japanese director Hideo Nakata, The Ring Two upsets categories like remake and sequel. Running below the sinuous narrative and generic entanglements of adaptation, translation, and sequelization is a pattern of shifting authorship. By analyzing the
discourse of authorship in industrial texts such as trade journals, newspaper articles, press kits, and DVD featurettes, this article argues that the logic of shifting authorship reflects Hollywood's flexible accumulation of international content and labor. The fetish of the original, discussed
and reinterpreted continuously in each subsequent installment of the Ringu/Ring franchise, becomes the basis for self-mythologizing and justification for Hollywood's new international division of cultural labor. Under these circumstances, Nakata's auteur status serves as (multi)cultural capital,
while his labor serves to ventriloquize Hollywood horror conventions and the style of director Gore Verbinski, whose presence continues to haunt the franchise as it is further passed along.
Review
Authors: Tubrett Dion
Published: December 2010

Horror Zone: The Cultural Experience of Contemporary Horror Cinema, Edited by Ian Conrich, (2010) London: I.B. Tauris, 320pp., ISBN: 9781848851511, Paperback, 51.50