bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enrichment and analysis of secretory lysosomes from lymphocyte populations
Authors: Schmidt Hendrik , Gelhaus Christoph , Lucius Ralph , Nebendahl Melanie , Leippe Matthias , Janssen Ottmar 
Published: July 2009

AbstractBackgroundIn specialized cells, such as mast cells, macrophages, T lymphocytes and Natural Killer cells in the immune system and for instance melanocytes in the skin, secretory lysosomes (SL) have evolved as bifunctional organelles that combine degradative and secretory properties. Mutations in lysosomal storage, transport or sorting molecules are associated with severe immunodeficiencies, autoimmunity and (partial) albinism. In order to analyze the function and content of secretory lysosomes in different cell populations, an efficient enrichment of these organelles is mandatory.ResultsBased on a combination of differential and density gradient centrifugation steps, we provide a protocol to enrich intact SL from expanded hematopoietic cells, here T lymphocytes and Natural Killer cells. Individual fractions were initially characterized by Western blotting using antibodies against an array of marker proteins for intracellular compartments. As indicated by the presence of LAMP-3 (CD63) and FasL (CD178), we obtained a selective enrichment of SL in one of the resulting organelle fractions. The robustness and reproducibility of the applied separation protocol was examined by a high-resolution proteome analysis of individual SL preparations of different donors by 2D difference gel electrophoresis (2D-DIGE).ConclusionThe provided protocol is readily applicable to enrich and isolate intact secretory vesicles from individual cell populations. It can be used to compare SL of normal and transformed cell lines or primary cell populations from healthy donors and patients with lysosomal storage or transport diseases, or from corresponding mutant mice. A subsequent proteome analysis allows the characterization of molecules involved in lysosomal maturation and cytotoxic effector function at high-resolution.
Constitutive RB1 mutation in a child conceived by in vitrofertilization: implications for genetic counseling
Authors: Barbosa Raquel H , Vargas Fernando R , Lucena Evandro , Bonvicino Cibele R , Seuánez Héctor N 
Published: July 2009

Abstract

Background
The purpose of this study was to identify mutations associated with bilateral retinoblastoma in a quadruplet conceived by in vitro fertilization, and to trace the parental origin of mutations in the four quadruplets and their father.

Methods
Mutational screening was carried out by sequencing. Genotyping was carried out for determining quadruplet zygosity.

Results
The proband was a carrier of a novel RB1 constitutive mutation (g.2056C>G) which was not detected in her father or her unaffected sisters, and of two other mutations (g.39606 C>T and g.174351T>A) also present in two monozygotic sisters. The novel mutation probably occurred de novo while the others were of likely maternal origin. The novel mutation, affecting the Kozak consensus at the 5'UTR of RB1 and g.174351T>A were likely associated to retinoblastoma in the proband.

Conclusion
Molecular diagnosis of retinoblastoma requires genotypic data of the family for determining hereditary transmission. In the case of children generated by IVF with oocytes from an anonymous donor which had been stored in a cell repository, this might not be successfully accomplished, making precise diagnosis impracticable for genetic counseling.
Role of KCNMA1gene in breast cancer invasion and metastasis to brain
Authors: Khaitan Divya , Sankpal Umesh T , Weksler Babette , Meister Edward A , Romero Ignacio A , Couraud Pierre-Olivier , Ningaraj Nagendra S 
Published: July 2009

Abstract

Background
The prognosis for patients with breast tumor metastases to brain is extremely poor. Identification of prognostic molecular markers of the metastatic process is critical for designing therapeutic modalities for reducing the occurrence of metastasis. Although ubiquitously present in most human organs, large-conductance calcium- and voltage-activated potassium channel (BKCa) channels are significantly upregulated in breast cancer cells. In this study we investigated the role of KCNMA1 gene that encodes for the pore-forming α-subunit of BKCa channels in breast cancer metastasis and invasion.

Methods
We performed Global exon array to study the expression of KCNMA1 in metastatic breast cancer to brain, compared its expression in primary breast cancer and breast cancers metastatic to other organs, and validated the findings by RT-PCR. Immunohistochemistry was performed to study the expression and localization of BKCa channel protein in primary and metastatic breast cancer tissues and breast cancer cell lines. We performed matrigel invasion, transendothelial migration and membrane potential assays in established lines of normal breast cells (MCF-10A), non-metastatic breast cancer (MCF-7), non-brain metastatic breast cancer cells (MDA-MB-231), and brain-specific metastatic breast cancer cells (MDA-MB-361) to study whether BKCa channel inhibition attenuates breast tumor invasion and metastasis using KCNMA1 knockdown with siRNA and biochemical inhibition with Iberiotoxin (IBTX).

Results
The Global exon array and RT-PCR showed higher KCNMA1 expression in metastatic breast cancer in brain compared to metastatic breast cancers in other organs. Our results clearly show that metastatic breast cancer cells exhibit increased BKCa channel activity, leading to greater invasiveness and transendothelial migration, both of which could be attenuated by blocking KCNMA1.

Conclusion
Determining the relative abundance of BKCa channel expression in breast cancer metastatic to brain and the mechanism of its action in brain metastasis will provide a unique opportunity to identify and differentiate between low grade breast tumors that are at high risk for metastasis from those at low risk for metastasis. This distinction would in turn allow for the appropriate and efficient application of effective treatments while sparing patients with low risk for metastasis from the toxic side effects of chemotherapy.
Prevalence and changes in chronic diseases among South Korean farmers: 1998 to 2005
Authors: Cha Eun Shil , Kong Kyoung Ae , Moon Eun Kyeong , Lee Won Jin 
Published: July 2009

Abstract

Background
Epidemiologic studies have suggested a unique pattern of disease among farmers in Western countries, but limited information is available about the magnitude of disease prevalence and their changes over time in Asian farmers. The aim of this study was to compare the prevalence and changes in chronic diseases among farmers with those of other occupational groups in South Korea.

Methods
Using data from three consecutive cross-sectional national surveys: the Korean National Health and Nutrition Examination Survey 1998 (n = 39,060), 2001 (n = 37,769), and 2005 (n = 34,145), we calculated age and gender-standardized prevalence of chronic diseases by the direct method and compared the prevalence changes from 1998 to 2005.

Results
Female farmers had significantly higher chronic disease prevalence than other occupational groups in all three surveys. Arthritis was the most prevalent chronic disease among farmers for both men and women. Compared with other populations, farmers demonstrated a higher prevalence of arthritis and intervertebral disc disorders. Farmers showed higher prevalence changes for intervertebral disc disorders than other occupational workers.

Conclusion
Our findings support that South Korean farmers have a distinct pattern of diseases prevalence from other populations. More detailed studies investigating the risk of musculoskeletal diseases and intensive intervention efforts to reduce the prevalence these diseases, particularly among female farmers, are required.
Worry as a window into the lives of people who use injection drugs: a factor analysis approach
Authors: Exner Heidi , Gibson Erin K , Stone Ryan , Lindquist Jennifer , Cowen Laura , Roth Eric A 
Published: July 2009

Abstract

Background
The concept of risk dominates the HIV/AIDS literature pertaining to People Who Use Injection Drugs (PWUID). In contrast the associated concept of worry is infrequently applied, even though it can produce important perspectives of PWUID's lives. This study asked a sample (n = 105) of PWUID enrolled in a Victoria, British Columbia needle exchange program to evaluate their degree of worry about fourteen factors they may encounter in their daily lives.

Methods
Exploratory factor analysis was used to analyze their responses.

Results
Factor analysis delineated three factors: 1) overall personal security, 2) injection drug use-specific risks including overdosing and vein collapse and, 3) contracting infectious diseases associated with injection drug use (e.g. HIV/AIDS and hepatitis C).

Conclusion
PWUID in this study not only worry about HIV/AIDS but also about stressful factors in their daily life which have been linked to both increased HIV/AIDS risk behaviour and decreased anti-retroviral treatment adherence. The importance PWUID give to this broad range of worry/concerns emphasizes the need to place HIV/AIDS intervention, education, and treatment programs within a broader harm-reduction framework that incorporates their perspectives on both worry and risk.
Evaluation of the impact of fibromyalgia on patients' sleep and the content validity of two sleep scales
Authors: Martin Susan , Chandran Arthi , Zografos Laurie , Zlateva Gergana 
Published: July 2009

Abstract

Background
Disturbed sleep is commonly reported in fibromyalgia (FM). Both the Sleep Quality Numeric Rating Scale (NRS) and the Medical Outcomes Study Sleep Scale (MOS-Sleep) have demonstrated positive psychometric properties in patients with FM. However, these assessments were developed prior to the current recommendation to include patient input during the concept elicitation or item generation phases. Therefore, the objective of this study was to evaluate the impact of FM on participants, including their sleep, and to test the content validity of these two sleep measures in FM patients.

Methods
Qualitative interviews were conducted in Raleigh, North Carolina and Detroit, Michigan with 20 adults who reported a physician diagnosis of FM. Sixteen participants were female, 13 were white, and the average age was 50 years. Two researchers conducted all interviews using a structured guide.

Results
Participants consistently reported that FM had a debilitating impact on their lives and their sleep, particularly getting to sleep and staying asleep. Participants responded positively to the Sleep Quality NRS as an assessment of their sleep. The majority of participants stated that they would not change the response numbering or wording of the item's anchors. Participants also responded positively to the 24-hour recall period of the Sleep Quality NRS. Participants found the 12-item MOS-Sleep to be appropriate and relevant; 19 participants indicated the measure captured all of their sleep-related symptoms. However, areas for potential modification were identified, such as the need to separate the item regarding awakening short of breath and awakening with a headache into two separate questions. Participants also questioned the relevance of the snoring and awakening short of breath items to FM. Half of participants expressed a preference for a daily rather than a weekly recall period.

Conclusion
This study demonstrates the significant impact that FM has on patients' lives, particularly sleep. While patients with FM were not part of the development of the generic sleep assessments that were evaluated, this study provides evidence of their content validity, supporting their use in FM studies. Modifications to the MOS-Sleep may improve the psychometric properties and relevance to patients with FM.
Vasoprotective effects of human CD34+ cells: towards clinical applications
Authors: Kiernan Thomas J , Boilson Barry A , Witt Tyra A , Dietz Allan B , Lerman Amir , Simari Robert D 
Published: July 2009

Abstract

Background
The development of cell-based therapeutics for humans requires preclinical testing in animal models. The use of autologous animal products fails to address the efficacy of similar products derived from humans. We used a novel immunodeficient rat carotid injury model in order to determine whether human cells could improve vascular remodelling following acute injury.

Methods
Human CD34+ cells were separated from peripheral buffy coats using automatic magnetic cell separation. Carotid arterial injury was performed in male Sprague-Dawley nude rats using a 2F Fogarty balloon catheter. Freshly harvested CD34+ cells or saline alone was administered locally for 20 minutes by endoluminal instillation. Structural and functional analysis of the arteries was performed 28 days later.

Results
Morphometric analysis demonstrated that human CD34+ cell delivery was associated with a significant reduction in intimal formation 4 weeks following balloon injury as compared with saline (I/M ratio 0.79 ± 0.18, and 1.71 ± 0.18 for CD34, and saline-treated vessels, respectively P < 0.05). Vasoreactivity studies showed that maximal relaxation of vessel rings from human CD34+ treated animals was significantly enhanced compared with saline-treated counterparts (74.1 ± 10.2, and 36.8 ± 12.1% relaxation for CD34+ cells and saline, respectively, P < 0.05)

Conclusion
Delivery of human CD34+ cells limits neointima formation and improves arterial reactivity after vascular injury. These studies advance the concept of cell delivery to effect vascular remodeling toward a potential human cellular product.
Immunization with an immunodominant self-peptide derived from glucose-6-phosphate isomerase induces arthritis in DBA/1 mice
Authors: Bruns Lisa , Frey Oliver , Morawietz Lars , Landgraf Christiane , Volkmer Rudolf , Kamradt Thomas 
Published: July 2009

Abstract
Introduction
T-helper (Th) lymphocytes are critically required for the pathogenesis of glucose-6-phosphate isomerase (G6PI)-induced arthritis, but neither the G6PI epitopes recognized by arthritogenic T cells nor their pathogenic effector functions have been fully elucidated to date. We aimed at identifying arthritogenic G6PI peptides.

Methods
We used a library of overlapping peptides spanning the entire G6PI sequence to identify the epitopes recognized by G6PI-specific Th cells. Immunodominant peptides were then used to immunize mice. Arthritis development was evaluated clinically and histologically. The humoral and cellular immune responses upon peptide immunization were analyzed by ELISA and multiparameter flow cytometry, respectively.

Results
We identified six immunodominant T-cell epitopes in DBA/1 mice, of which three are arthritogenic. One of these peptides (G6PI469–483) is identical in man and mice. Immunization with this peptide induces arthritis, which is less severe and of shorter duration than arthritis induced by immunization with full-length G6PI. Upon immunization with either G6PI or peptide, the antigen-specific Th cells produce IL-17, RANKL, IFNγ and TNFα.

Conclusions
We identified immunodominant and arthritogenic epitopes of G6PI. Not all immunodominant peptides are arthritogenic. This is the first description of arthritis induced by immunization with a self-peptide in mice.
Neutrophil antimicrobial defense against Staphylococcus aureus is mediated by phagolysosomal but not extracellular trap-associated cathelicidin
Authors: Jann Naja J , Schmaler Mathias , Kristian Sascha A , Radek Katherine A , Gallo Richard L , Nizet Victor , Peschel Andreas , Landmann Regine 
Published: July 2009

Abstract
Study took advantage of the hyper-susceptible phenotype of S. aureus ΔdltA against cationic AMPs to investigate the impact of the murine cathelicidin CRAMP to identify its key site of action in neutrophils.
Neutrophils kill invading pathogens by AMPs, including cathelicidins, ROS, and NETs. The human pathogen Staphylococcus aureus exhibits enhanced resistance to neutrophil AMPs, including the murine cathelicidin CRAMP, in part, as a result of alanylation of teichoic acids by the dlt operon. In this study, we took advantage of the hypersusceptible phenotype of S. aureus ΔdltA against cationic AMPs to study the impact of the murine cathelicidin CRAMP on staphylococcal killing and to identify its key site of action in murine neutrophils. We demonstrate that CRAMP remained intracellular during PMN exudation from blood and was secreted upon PMA stimulation. We show first evidence that CRAMP was recruited to phagolysosomes in infected neutrophils and exhibited intracellular activity against S. aureus. Later in infection, neutrophils produced NETs, and immunofluorescence revealed association of CRAMP with S. aureus in NETs, which similarly killed S. aureus wt and ΔdltA, indicating that CRAMP activity was reduced when associated with NETs. Indeed, the presence of DNA reduced the antimicrobial activity of CRAMP, and CRAMP localization in response to S. aureus was independent of the NADPH oxidase, whereas killing was partially dependent on a functional NADPH oxidase. Our study indicates that neutrophils use CRAMP in a timed and locally coordinated manner in defense against S. aureus.
Host-pathogen interactions during coronavirus infection of primary alveolar epithelial cells
Authors: Miura Tanya A , Holmes Kathryn V 
Published: July 2009

Abstract
Innate immune responses in coronavirus infections of the respiratory tract are analyzed in primary differentiated airway and alveolar epithelial cells.
Viruses that infect the lung are a significant cause of morbidity and mortality in animals and humans worldwide. Coronaviruses are being associated increasingly with severe diseases in the lower respiratory tract. Alveolar epithelial cells are an important target for coronavirus infection in the lung, and infected cells can initiate innate immune responses to viral infection. In this overview, we describe in vitro models of highly differentiated alveolar epithelial cells that are currently being used to study the innate immune response to coronavirus infection. We have shown that rat coronavirus infection of rat alveolar type I epithelial cells in vitro induces expression of CXC chemokines, which may recruit and activate neutrophils. Although neutrophils are recruited early in infection in several coronavirus models including rat coronavirus. However, their role in viral clearance and/or immune-mediated tissue damage is not understood. Primary cultures of differentiated alveolar epithelial cells will be useful for identifying the interactions between coronaviruses and alveolar epithelial cells that influence the innate immune responses to infection in the lung. Understanding the molecular details of these interactions will be critical for the design of effective strategies to prevent and treat coronavirus infections in the lung.
clueless, a conserved Drosophila gene required for mitochondrial subcellular localization, interacts genetically withparkin
Authors: Cox Rachel T. , Spradling Allan C. 
Published: July 2009

SUMMARYParkinson’s disease has been linked to altered mitochondrial function. Mutations in parkin (park), the Drosophila ortholog of a human gene that is responsible for many familial cases of Parkinson’s disease, shorten life span, abolish fertility and disrupt mitochondrial structure. However, the role played by Park in mitochondrial function remains unclear. Here, we describe a novel Drosophila gene, clueless (clu), which encodes a highly conserved tetratricopeptide repeat protein that is related closely to the CluA protein of Dictyostelium, Clu1 of Saccharomyces cerevisiae and to similar proteins in diverse metazoan eukaryotes from Arabidopsis to humans. Like its orthologs, loss of Drosophila clu causes mitochondria to cluster within cells. We find that strong clu mutations resemble park mutations in their effects on mitochondrial function and that the two genes interact genetically. Conversely, mitochondria in park homozygotes become highly clustered. We propose that Clu functions in a novel pathway that positions mitochondria within the cell based on their physiological state. Disruption of the Clu pathway may enhance oxidative damage, alter gene expression, cause mitochondria to cluster at microtubule plus ends, and lead eventually to mitochondrial failure.
Review of diesel emissions and control
Authors: Johnson T V 
Published: July 2009

This summary covers key and representative developments in diesel emissions regulations, engine developments, and nitrogen oxide (NO x), particulate matter (PM), and hydrocarbon (HC) remediation. In California discussions are taking place on another round of criteria pollutant tightening on light duty. Carbon dioxide (CO2) regulations are being discussed in the developed markets. The US mandates on fuel economy and the proposed standards in Europe and California will result in a paradigm shift, wherein technologies previously rejected based on customer value will now be re-evaluated. Heavy-duty regulatory developments are now focused on Europe, where Euro VI regulations have been proposed for about 2013. The regulations are intended to be technology harmonizing with the US and Japanese regulations. Engines are continuing to make very impressive progress, with clean combustion strategies in active development mainly for US light-duty application. Heavy-duty research engines are more focused on traditional approaches, and will provide numerous engine/aftertreatment options for hitting the tight US 2010 regulations. NO x control is focusing on selective catalytic reduction (SCR) for diverse applications. Zeolite catalysts will be the mainstay for Japan, US, and Euro VI applications. The focus is catalyst improvements and system optimization. Lean NO x traps (LNTs) are effective up to about 70–80 per cent deNO x efficiency, and are being used for light-duty applications with significant improvements coming on precious metal utilization. Diesel particulate filter (DPF) technology is in a state of optimization and cost reduction. Very sophisticated management strategies are being utilized, which open up options for new filter materials and alternative system architectures. New catalyst formulations that oxidize soot at the catalyst—soot interface are emerging and are enhancing regeneration in the first commercial applications. Diesel oxidation catalysts (DOCs) are lighting-off and generating more NO2 for DPF regeneration at lower temperatures. For a more comprehensive review of these technologies the reader is directed to Johnson, T. V., Diesel emissions in review, SAE 2008-01-0121.
The Absent Elegy: Performing Trauma in The Winter's Tale
Authors: Sherman Donovan 
Published: July 2009

This article addresses the current intersection of literary- and performance-based studies of Shakespeare's plays through the lens of The Winter's Tale . It reads the play as a complex and profound exploration of the interplay, and possible incommensurability, of performance and text in which the boundaries between the theatrical and literary event are broken down, serving as a marker for the study of Renaissance literature in general. Drawing on the field of trauma theory, the article traces the development of Leontes's crisis of ontological and subjective coherence, positing it as ingrained in the play's negotiation of the border between performance and text. A focus on The Winter's Tale's narrative preoccupation with the infliction of and attempted recovery from an event of loss affords a reading of the underexamined figure of Leontes's son, Mamillius, as an unstable figure of narrative re-inscription and recovery, who disturbs, in an emphatically theatrical way, the logic of the play through his embodiment of loss itself. By positioning this figure in conjunction with the play's climactic 'statue' scene, the article reveals the play's presentation of a distinctly posttraumatic process, in which, through the statue's reconciliation of signifier and referent, it overwrites its own theatricality with new systems of literary legibility. In doing so, the article argues, The Winter's Tale offers a question of the constantly intermingling modes of performance and literature.