bluuebunny/crossref_metadata_2025 huggingface dataset:
Alcohol, ageing and dementia: A Scottish perspective
Authors: McCabe Louise F.
Published: March 2011

There are complex relationships between alcohol use; processes of ageing both social and biological; and cognitive impairment. This paper takes Scotland as a case study to begin to unpick and explore these relationships and their implications for people who drink as they age. In recent years several policy documents have been published in Scotland that acknowledge the increasing prevalence of alcohol problems among people as they age. This paper is based on a review of the literature, analysis of recent Scottish policy documents and two research projects undertaken by the author. The links between alcohol use and dementia are discussed, focusing on biological and social implications for individuals as they age. Current responses in Scotland to these different issues are examined from individual, cultural and policy perspectives. Despite efforts in Scotland the needs of ageing and older individuals who experience alcohol-related morbidity are not sufficiently addressed.
Development of a Theory-Based (PEN-3 and Health Belief Model), Culturally Relevant Intervention on Cervical Cancer Prevention Among Latina Immigrants Using Intervention Mapping
Authors: Scarinci Isabel C. , Bandura Lisa , Hidalgo Bertha , Cherrington Andrea
Published: March 2011

The development of efficacious theory-based, culturally relevant interventions to promote cervical cancer prevention among underserved populations is crucial to the elimination of cancer disparities. The purpose of this article is to describe the development of a theory-based, culturally relevant intervention focusing on primary (sexual risk reduction) and secondary (Pap smear) prevention of cervical cancer among Latina immigrants using intervention mapping (IM). The PEN-3 and Health Belief Model provided theoretical guidance for the intervention development and implementation. IM provides a logical five-step framework in intervention development: delineating proximal program objectives, selecting theory-based intervention methods and strategies, developing a program plan, planning for adoption in implementation, and creating evaluation plans and instruments. We first conducted an extensive literature review and qualitatively examined the sociocultural factors associated with primary and secondary prevention of cervical cancer. We then proceeded to quantitatively validate the qualitative findings, which led to development matrices linking the theoretical constructs with intervention objectives and strategies as well as evaluation. IM was a helpful tool in the development of a theory-based, culturally relevant intervention addressing primary and secondary prevention among Latina immigrants.
Disaster Preparedness of Linguistically Isolated Populations
Authors: Nepal Vishnu , Banerjee Deborah , Perry Mark , Scott Deborah
Published: March 2011

In the absence of culturally and linguistically appropriate disaster preparedness plans, several linguistically isolated and culturally diverse population groups are disproportionately disadvantaged in the United States. The communication gap poses challenges to emergency preparedness planners and response personnel in predisaster communication and postdisaster response efforts. Houston Department of Health and Human Services aimed to develop practical recommendations for local emergency response personnel so as to improve dissemination of emergency information and equitable delivery of services to linguistically isolated communities in the greater Houston area. Sixteen focus group discussions were conducted among linguistically isolated immigrant populations living in the greater Houston metropolitan area who primarily spoke one of the Spanish, Chinese, Vietnamese, and Somali languages. Our questions focused on general knowledge and understanding of disasters and explored experiences during Houston’s most recent disaster, Hurricane Ike. We found that (a) understanding of disaster and preparedness is contextual, (b) awareness of preparedness needs and actual plans among LIPs is inadequate, and (c) word of mouth is the preferred information source for linguistically isolated groups. Disaster preparedness plans of a given jurisdiction should reflect the culturally and linguistically appropriate components addressing the needs, concerns, context-based knowledge or awareness, and perceptions of linguistically isolated populations.
The Implementation of a Statewide Bullying Prevention Program
Authors: Schroeder Betsy A. , Messina Allison , Schroeder Diana , Good Karla , Barto Shiryl , Saylor Jennifer , Masiello Matthew
Published: March 2011

Bullying in schools has become recognized as a significant public health problem. The Olweus Bullying Prevention Program (OBPP) has been identified as an effective means to reduce bullying behavior in schools. The goal of this large population-based initiative was to reduce bullying by producing a quantifiable change in school climate using an established program and standardized measurement tool. Program participants over a 2-year period included 56,137 students and more than 2,400 teachers from 107 schools in 49 counties across Pennsylvania. An age cohorts design was used, and data from two equivalent age cohorts of students were compared at two or more points in time. After 1 to 2 years of program implementation, across cohorts, there were reductions in student self-reports of bullying others, and improvements in student perceptions of adults’ responsiveness, and students’ attitudes about bullying. This study is the largest bullying prevention initiative to date in the United States. This initiative reaffirms the efficacy of the OBPP, emphasizes the importance of an identified coalition, and highlights several positive outcomes. It is recommended that the OBPP be implemented through the establishment of community partnerships and coalitions as consistent with the public health model.
Implementing School Health Promotion in Austria
Authors: Flaschberger Edith , Nitsch Martina , Waldherr Karin
Published: March 2011

A pilot training course on school health promotion in Austria focused on supporting teachers and school heads during the implementation of the health-promoting school (HPS) concept. The participants talked about their experiences during the implementation phase in focus groups held in a feedback seminar at the end of the training course. The focus groups were analyzed by using an inductive approach. A coding method for thematic analysis was applied. Compatibility of the training course with the school setting was examined and supporting and hindering structures and processes pertaining to the schools and the training course are described. The results can be displayed in a model showing connections between the themes. Internal and external collaboration structures were identified as central aspects in the implementation phase. Particularly collaboration within the faculty is shown to be of importance when implementing the HPS approach. Voluntary cooperation especially of teachers, which is often connected with work in their spare time, is another aspect influencing the sustainability and therefore also the compatibility of the training course with the school setting. For future designs of teacher training courses in health promotion, the aspects of collaboration, teamwork, and collective commitment have to be taken into consideration.
Professionalization and the Experience-Based Expert: Strengthening Partnerships Between Health Educators and Community Health Workers
Authors: Gilkey Melissa , Garcia Cezanne Cochran , Rush Carl
Published: March 2011

The recent inclusion of community health workers (CHWs) in the U.S. Department of Labor’s 2010 Standard Occupational Classification System provides an opportunity for health educators to reflect on their relationship with CHWs. The authors discuss the ways that health educators and CHWs differ in their orientation toward professionalization and employ the concept of the “experience-based expert” to highlight what they believe to be the unique contributions of CHWs. Finally, considerations important for health educators and CHWs as they work to advance supportive and complementary practices are discussed.
Long-term risk factor management after inpatient cardiac rehabilitation by means of a structured post-care programme
Authors: Harb Birgit M , Wonisch Manfred , Brandt Dieter , Müller Rudolf
Published: March 2011

Background: Initiation of a long-term improvement of cardiac risk factors is one of the major aims of a cardiac rehabilitation/secondary prevention programme.

Methods and participants: The Health Guide collected data in terms of cardiac risk factors: blood pressure, resting pulse, total cholesterol, LDL cholesterol, body weight, physical activity and number of cigarettes at admission and discharge after a stationary rehabilitation programme and every 3 months. After 12 months the Health Guides were returned. In the prospective study 2664 patients (71.8% men, age: MV = 62.94 years, SD = 9.96; 28% women, MV = 67.59 years, SD = 9.53) with coronary heart disease (CHD) were included.

Results: All cardiac risk factors documented by the Health Guide improved during the cardiac rehabilitation programme. After one year, risk factors were significantly lower than at admission, apart from total cholesterol. The individual goal in terms of body weight and LDL cholesterol was partially achieved after the rehabilitation programme and maintained after one year. In the investigation years, 2004–2007, the cholesterol and blood pressure were significantly lower than in the years 2000–2003.

Conclusion: The use of a Health Guide resulted in an improved long-term effect of a cardiac rehabilitation/secondary prevention programme. It is a simple and cheap intervention and can help in the guidance of the patients.
The Link between Educational Expectations and Effort in the College-for-all Era
Authors: Domina Thurston , Conley AnneMarie , Farkas George
Published: March 2011

From the Wisconsin status attainment model to rational choice, classical sociological, social-psychological, and economic theories of student educational transitions have assumed that students’ expectations are positively related to their ultimate attainment. However, the growth of the college-for-all ethos raises questions about that assumption. Noting that American students’ educational expectations rapidly outpaced their educational attainments, Rosenbaum (2001 ) argues that increasingly unrealistic expectations have perverse negative effects on the school engagement of American high school students. In this article, the authors test the relationship between student expectations and effort using data from a unique longitudinal study of student motivation and three national cohort studies. Contrary to the college-for-all critique, the authors find that educational expectations continue to have robust positive effects on student perceptions regarding the future utility of high school academics and student effort in high school. The relationship between expectations and effort is somewhat attenuated for very low-achieving students and it is weaker today than it was in 1980. Nonetheless, the authors’ analyses indicate that the expansion of college expectations has had a net positive effect on American high school students’ effort.
MUC1-C oncoprotein suppresses reactive oxygen species–induced terminal differentiation of acute myelogenous leukemia cells
Authors: Yin Li , Wu Zekui , Avigan David , Rosenblatt Jacalyn , Stone Richard , Kharbanda Surender , Kufe Donald
Published: March 2011

Acute myeloid leukemia (AML) cells are characterized by unlimited self-renewal and an impaired capacity to undergo terminal differentiation. The MUC1 oncoprotein is aberrantly expressed in AML cells; however, there has been no evidence for involvement of MUC1 in myeloid leukemogenesis. Cell-penetrating peptide inhibitors of the MUC1-C subunit block its oligomerization and thereby oncogenic function. The present results demonstrate that treatment of human MOLM-14 and MV4-11 AML cells with these inhibitors is associated with arrest of growth, induction of late apoptosis/necrosis, and loss of self-renewal capacity. Similar results were obtained with primary blasts from patients with AML. Inhibition of MUC1-C was associated with increases in reactive oxygen species (ROS) and depletion of glutathione. Increases in ROS have been linked to induction of hematopoietic cell differentiation along the myeloid lineage. In this regard, inhibition of MUC1-C was associated with induction of a terminally differentiated myeloid phenotype in AML cell lines and primary blasts by an ROS-dependent mechanism. These findings indicate that MUC1-C function is of importance to AML cell self-renewal and that inhibition of MUC1-C represents a potential therapeutic approach to induce terminal differentiation of AML cells.
From an old remedy to a magic bullet: molecular mechanisms underlying the therapeutic effects of arsenic in fighting leukemia
Authors: Chen Sai-Juan , Zhou Guang-Biao , Zhang Xiao-Wei , Mao Jian-Hua , de Thé Hugues , Chen Zhu
Published: March 2011

Arsenic had been used in treating malignancies from the 18th to mid-20th century. In the past 3 decades, arsenic was revived and shown to be able to induce complete remission and to achieve, when combined with all-trans retinoic acid and chemotherapy, a 5-year overall survival of 90% in patients with acute promyelocytic leukemia driven by the t(15;17) translocation-generated promyelocytic leukemia–retinoic acid receptor α (PML-RARα) fusion. Molecularly, arsenic binds thiol residues and induces the formation of reactive oxygen species, thus affecting numerous signaling pathways. Interestingly, arsenic directly binds the C3HC4 zinc finger motif in the RBCC domain of PML and PML-RARα, induces their homodimerization and multimerization, and enhances their interaction with the SUMO E2 conjugase Ubc9, facilitating subsequent sumoylation/ubiquitination and proteasomal degradation. Arsenic-caused intermolecular disulfide formation in PML also contributes to PML-multimerization. All-trans retinoic acid, which targets PML-RARα for degradation through its RARα moiety, synergizes with arsenic in eliminating leukemia-initiating cells. Arsenic perturbs a number of proteins involved in other hematologic malignancies, including chronic myeloid leukemia and adult T-cell leukemia/lymphoma, whereby it may bring new therapeutic benefits. The successful revival of arsenic in acute promyelocytic leukemia, together with modern mechanistic studies, has thus allowed a new paradigm to emerge in translational medicine.
HIV-specific CD8+ T cells from HIV+ individuals receiving HAART can be expanded ex vivo to augment systemic and mucosal immunity in vivo
Authors: Chapuis Aude G. , Casper Corey , Kuntz Steve , Zhu Jia , Tjernlund Annelie , Diem Kurt , Turtle Cameron J. , Cigal Melinda L. , Velez Roxanne , Riddell Stanley , Corey Lawrence , Greenberg Philip D.
Published: March 2011

Most HIV+ individuals require lifelong highly active antiretroviral therapy (HAART) to suppress HIV replication, but fail to eliminate the virus in part because of residual replication in gut-associated lymphoid tissues (GALT). Naturally elicited HIV-specific CD8+ T cells generated in the acute and chronic infectious phases exhibit antiviral activity, but decrease in number after HAART. Therapeutic vaccines represent a potential strategy to expand cellular responses, although previous efforts have been largely unsuccessful, conceivably because of a lack of responding HIV-specific central-memory CD8+ T cells (Tcm). To determine whether patients receiving HAART possess CD8+ T cells with Tcm qualities that are amenable to augmentation, HIV-specific CD8+ T-cell clones were derived from HIV-reactive CD28+CD8+ T-cell lines isolated from 7 HIV+ HAART-treated patients, expanded ex vivo, and reinfused into their autologous host. Tracking of the cells in vivo revealed that clones could persist for ≥ 84 days, maintain expression and/or re-express CD28, up-regulate CD62L, secrete IL-2, proliferate on cognate Ag encounter and localize to the rectal mucosa. These results suggest some infused cells exhibited phenotypic and functional characteristics shared with Tcm in vivo, and imply that more effective therapeutic vaccination strategies targeting CD8+ Tcm in patients on HAART might provide hosts with expanded, long-lasting immune responses not only systemically but also in GALT. This study is registered at www.clinicaltrials.gov as NCT00110578.
KL-6 concentration in pulmonary epithelial lining fluid is a useful prognostic indicator in patients with acute respiratory distress syndrome
Authors: Kondo Tomohiro , Hattori Noboru , Ishikawa Nobuhisa , Murai Hiroshi , Haruta Yoshinori , Hirohashi Nobuyuki , Tanigawa Koichi , Kohno Nobuoki
Published: March 2011

Background

KL-6 is a mucin-like glycoprotein expressed on the surface of alveolar type II cells. Elevated concentrations of KL-6 in serum and epithelial lining fluid (ELF) in patients with acute respiratory distress syndrome (ARDS) have been previously reported; however, kinetics and prognostic significance of KL-6 have not been extensively studied. This study was conducted to clarify these points in ARDS patients.

Methods

Thirty-two patients with ARDS who received mechanical ventilation under intubation were studied for 28 days. ELF and blood were obtained from each patient at multiple time points after the diagnosis of ARDS. ELF was collected using a bronchoscopic microsampling procedure, and ELF and serum KL-6 concentrations were measured.

Results

KL-6 levels in ELF on days 0 to 3 after ARDS diagnosis were significantly higher in nonsurvivors than in survivors, and thereafter, there was no difference in concentrations between the two groups. Serum KL-6 levels did not show statistically significant differences between nonsurvivors and survivors at any time point. When the highest KL-6 levels in ELF and serum sample from each patient were examined, KL-6 levels in both ELF and serum were significantly higher in nonsurvivors than in survivors. The optimal cut-off values were set at 3453 U/mL for ELF and 530 U/mL for serum by receiver operating characteristic (ROC) curve analyses. Patients with KL-6 concentrations in ELF higher than 3453 U/mL or serum concentrations higher than 530 U/mL had significantly lower survival rates up to 90 days after ARDS diagnosis.

Conclusions

ELF and serum KL-6 concentrations were found to be good indicators of clinical outcome in ARDS patients. Particularly, KL-6 levels in ELF measured during the early period after the diagnosis were useful for predicting prognosis in ARDS patients.
FGF signalling through RAS/MAPK and PI3K pathways regulates cell movement and gene expression in the chicken primitive streak without affecting E-cadherin expression
Authors: Hardy Katharine M , Yatskievych Tatiana A , Konieczka JH , Bobbs Alexander S , Antin Parker B
Published: March 2011

Background

FGF signalling regulates numerous aspects of early embryo development. During gastrulation in amniotes, epiblast cells undergo an epithelial to mesenchymal transition (EMT) in the primitive streak to form the mesoderm and endoderm. In mice lacking FGFR1, epiblast cells in the primitive streak fail to downregulate E-cadherin and undergo EMT, and cell migration is inhibited. This study investigated how FGF signalling regulates cell movement and gene expression in the primitive streak of chicken embryos.

Results

We find that pharmacological inhibition of FGFR activity blocks migration of cells through the primitive streak of chicken embryos without apparent alterations in the level or intracellular localization of E-cadherin. E-cadherin protein is localized to the periphery of epiblast, primitive streak and some mesodermal cells. FGFR inhibition leads to downregulation of a large number of regulatory genes in the preingression epiblast adjacent to the primitive streak, the primitive streak and the newly formed mesoderm. This includes members of the FGF, NOTCH, EPH, PDGF, and canonical and non-canonical WNT pathways, negative modulators of these pathways, and a large number of transcriptional regulatory genes. SNAI2 expression in the primitive streak and mesoderm is not altered by FGFR inhibition, but is downregulated only in the preingression epiblast region with no significant effect on E-cadherin. Furthermore, over expression of SNAIL has no discernable effect on E-cadherin protein levels or localization in epiblast, primitive streak or mesodermal cells. FGFR activity modulates distinct downstream pathways including RAS/MAPK and PI3K/AKT. Pharmacological inhibition of MEK or AKT indicate that these downstream effectors control discrete and overlapping groups of genes during gastrulation. FGFR activity regulates components of several pathways known to be required for cell migration through the streak or in the mesoderm, including RHOA, the non-canonical WNT pathway, PDGF signalling and the cell adhesion protein N-cadherin.

Conclusions

In chicken embryos, FGF signalling regulates cell movement through the primitive streak by mechanisms that appear to be independent of changes in E-cadherin expression or protein localization. The positive and negative effects on large groups of genes by pharmacological inhibition of FGF signalling, including major signalling pathways and transcription factor families, indicates that the FGF pathway is a focal point of regulation during gastrulation in chicken.
Organization and evolution of hsp70clusters strikingly differ in two species of Stratiomyidae (Diptera) inhabiting thermally contrasting environments
Authors: Garbuz David G , Yushenova Irina A , Zatsepina Olga G , Przhiboro Andrey A , Bettencourt Brian R , Evgen'ev Michael B
Published: March 2011

Background

Previously, we described the heat shock response in dipteran species belonging to the family Stratiomyidae that develop in thermally and chemically contrasting habitats including highly aggressive ones. Although all species studied exhibit high constitutive levels of Hsp70 accompanied by exceptionally high thermotolerance, we also detected characteristic interspecies differences in heat shock protein (Hsp) expression and survival after severe heat shock. Here, we analyzed genomic libraries from two Stratiomyidae species from thermally and chemically contrasting habitats and determined the structure and organization of their hsp70 clusters.

Results

Although the genomes of both species contain similar numbers of hsp70 genes, the spatial distribution of hsp70 copies differs characteristically. In a population of the eurytopic species Stratiomys singularior, which exists in thermally variable and chemically aggressive (hypersaline) conditions, the hsp70 copies form a tight cluster with approximately equal intergenic distances. In contrast, in a population of the stenotopic Oxycera pardalina that dwells in a stable cold spring, we did not find hsp70 copies in tandem orientation. In this species, the distance between individual hsp70 copies in the genome is very large, if they are linked at all. In O. pardalina we detected the hsp68 gene located next to a hsp70 copy in tandem orientation. Although the hsp70 coding sequences of S. singularior are highly homogenized via conversion, the structure and general arrangement of the hsp70 clusters are highly polymorphic, including gross aberrations, various deletions in intergenic regions, and insertion of incomplete Mariner transposons in close vicinity to the 3'-UTRs.

Conclusions

The hsp70 gene families in S. singularior and O. pardalina evolved quite differently from one another. We demonstrated clear evidence of homogenizing gene conversion in the S. singularior hsp70 genes, which form tight clusters in this species. In the case of the other species, O. pardalina, we found no clear trace of concerted evolution for the dispersed hsp70 genes. Furthermore, in the latter species we detected hsp70 pseudogenes, representing a hallmark of the birth-and-death process.
RefGenes: identification of reliable and condition specific reference genes for RT-qPCR data normalization
Authors: Hruz Tomas , Wyss Markus , Docquier Mylene , Pfaffl Michael W , Masanetz Sabine , Borghi Lorenzo , Verbrugghe Phebe , Kalaydjieva Luba , Bleuler Stefan , Laule Oliver , Descombes Patrick , Gruissem Wilhelm , Zimmermann Philip
Published: March 2011

Background

RT-qPCR is a sensitive and increasingly used method for gene expression quantification. To normalize RT-qPCR measurements between samples, most laboratories use endogenous reference genes as internal controls. There is increasing evidence, however, that the expression of commonly used reference genes can vary significantly in certain contexts.

Results

Using the Genevestigator database of normalized and well-annotated microarray experiments, we describe the expression stability characteristics of the transciptomes of several organisms. The results show that a) no genes are universally stable, b) most commonly used reference genes yield very high transcript abundances as compared to the entire transcriptome, and c) for each biological context a subset of stable genes exists that has smaller variance than commonly used reference genes or genes that were selected for their stability across all conditions.

Conclusion

We therefore propose the normalization of RT-qPCR data using reference genes that are specifically chosen for the conditions under study. RefGenes is a community tool developed for that purpose. Validation RT-qPCR experiments across several organisms showed that the candidates proposed by RefGenes generally outperformed commonly used reference genes. RefGenes is available within Genevestigator at http://www.genevestigator.com.
Separation of the bacterial species, Escherichia coli, from mixed-species microbial communities for transcriptome analysis
Authors: Dai Dongjuan , Holder Diane , Raskin Lutgarde , Xi Chuanwu
Published: March 2011

Background

The study of bacterial species interactions in a mixed-species community can be facilitated by transcriptome analysis of one species in the community using cDNA microarray technology. However, current applications of microarrays are mostly limited to single species studies. The purpose of this study is to develop a method to separate one species, Escherichia coli as an example, from mixed-species communities for transcriptome analysis.

Results

E. coli cells were separated from a dual-species (E. coli and Stenotrophomonas maltophilia) community using immuno-magnetic separation (IMS). High recovery rates of E. coli were achieved. The purity of E. coli cells was as high as 95.0% separated from suspended mixtures consisting of 1.1 - 71.3% E. coli, and as high as 96.0% separated from biofilms with 8.1% E. coli cells. Biofilms were pre-dispersed into single-cell suspensions. The reagent RNAlater (Ambion, Austin, TX) was used during biofilm dispersion and IMS to preserve the transcriptome of E. coli. A microarray study and quantitative PCR confirmed that very few E. coli genes (only about eight out of 4,289 ORFs) exhibited a significant change in expression during dispersion and separation, indicating that transcriptional profiles of E. coli were well preserved.

Conclusions

A method based on immuno-magnetic separation (IMS) and application of RNAlater was developed to separate a bacterial species, E. coli as an example, from mixed-species communities while preserving its transcriptome. The method combined with cDNA microarray analysis should be very useful to study species interactions in mixed-species communities.
The role of Endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) for qualitative diagnosis of mediastinal and hilar lymphadenopathy: a prospective analysis
Authors: Ye Ting , Hu Hong , Luo Xiaoyang , Chen Haiquan
Published: March 2011

Background

Recently EBUS-TBNA, which has a sensitivity of 94.6%, specificity of 100% and diagnostic accuracy rate of 96.3% as previously reported, has been widely used for patients with mediastinal and hilar lymphadenopathy or suspected lung cancer to get accurate diagnosis. The purpose of the current study was to evaluate the usefulness of EBUS-TBNA in obtaining cytological and histological diagnosis of mediastinal and hilar lymph nodes compared to the results obtained with conventional mediastinoscopy as previously reported, and to assess the relationship of diagnostic accuracy and number of passes and size of lymph nodes.

Methods

101 patients with mediastinal and hilar lymphadenopathy or suspected lung cancer in our institution were included in this prospective study. EBUS-TBNA was performed in all cases. The final diagnosis was confirmed by cytology, surgical results, and/or clinical follow-up for at least 6 months. Sensitivity, specificity, accuracy, and positive and negative predictive values were calculated using standard formulas.

Results

In 101 patients, EBUS-TBNA was successfully performed to obtain samples from 225 lymph nodes, 7 lung masses, 1 mediastinal mass and 2 esophageal masses. 63 malignant tumors and 38 benign diseases were confirmed. Epidermal growth factor receptor mutation was detected in 10 biopsy samples, and epidermal growth factor receptor mutation was detected in 4 cases. With respect to the correct diagnosis of mediastinal and hilar lymphadenopathy, EBUS-TBNA had a sensitivity of 95.08%, specificity of 100%, positive predictive value of 100%, negative predictive value of 93.02%, and overall accuracy of 97.02%. The relationship of diagnostic accuracy and number of lymph node passes or size of lymph nodes was both insignificant (p = 0.27; p = 0.23). The procedure was uneventful without complications.

Conclusions

EBUS-TBNA is an accurate and safe tool in diagnosis of mediastinal and hilar lymphadenopathy. It cannot completely replace mediastinoscopy, it may indeed reduce the number of mediastinoscopy procedures. In some cases, it can necessarily be the first-line procedure before mediastinoscopy.
Correlation of SHOX2 Gene Amplification and DNA Methylation in Lung Cancer Tumors
Authors: Schneider Katja U , Dietrich Dimo , Fleischhacker Michael , Leschber Gunda , Merk Johannes , Schäper Frank , Stapert Henk R , Vossenaar Erik R , Weickmann Sabine , Liebenberg Volker , Kneip Christoph , Seegebarth Anke , Erdogan Fikret , Rappold Gudrun , Schmidt Bernd
Published: March 2011

Background

DNA methylation in the SHOX2 locus was previously used to reliably detect lung cancer in a group of critical controls, including 'cytologically negative' samples with no visible tumor cell content, at a high specificity based on the analysis of bronchial lavage samples. This study aimed to investigate, if the methylation correlates with SHOX2 gene expression and/or copy number alterations. An amplification of the SHOX2 gene locus together with the observed tumor-specific hypermethylation might explain the good performance of this marker in bronchial lavage samples.

Methods

SHOX2 expression, gene copy number and DNA methylation were determined in lung tumor tissues and matched morphologically normal adjacent tissues (NAT) from 55 lung cancer patients. Quantitative HeavyMethyl (HM) real-time PCR was used to detect SHOX2 DNA methylation levels. SHOX2 expression was assayed with quantitative real-time PCR, and copy numbers alterations were measured with conventional real-time PCR and array CGH.

Results

A hypermethylation of the SHOX2 locus in tumor tissue as compared to the matched NAT from the same patient was detected in 96% of tumors from a group of 55 lung cancer patients. This correlated highly significantly with the frequent occurrence of copy number amplification (p < 0.0001), while the expression of the SHOX2 gene showed no difference.

Conclusions

Frequent gene amplification correlated with hypermethylation of the SHOX2 gene locus. This concerted effect qualifies SHOX2 DNA methylation as a biomarker for lung cancer diagnosis, especially when sensitive detection is needed, i.e. in bronchial lavage or blood samples.
Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield
Authors: Routledge Sarah J , Hewitt Christopher J , Bora Nagamani , Bill Roslyn M
Published: March 2011

Background

Pichia pastorisis a widely-used host for recombinant protein production. Initial screening for both suitable clones and optimum culture conditions is typically carried out in multi-well plates. This is followed by up-scaling either to shake-flasks or continuously stirred tank bioreactors. A particular problem in these formats is foaming, which is commonly prevented by the addition of chemical antifoaming agents. Intriguingly, antifoams are often added without prior consideration of their effect on the yeast cells, the protein product or the influence on downstream processes such as protein purification. In this study we characterised, for the first time, the effects of five commonly-used antifoaming agents on the total amount of recombinant green fluorescent protein (GFP) secreted from shake-flask cultures of this industrially-relevant yeast.

Results

Addition of defined concentrations of Antifoam A (Sigma), Antifoam C (Sigma), J673A (Struktol), P2000 (Fluka) or SB2121 (Struktol) to shake-flask cultures ofP. pastorisincreased the total amount of recombinant GFP in the culture medium (the total yield) and in the case of P2000, SB2121 and J673A almost doubled it. When normalized to the culture density, the GFP specific yield (μg OD595-1) was only increased for Antifoam A, Antifoam C and J673A. Whilst none of the antifoams affected the growth rate of the cells, addition of P2000 or SB2121 was found to increase culture density. There was no correlation between total yield, specific yield or specific growth rate and the volumetric oxygen mass transfer coefficient (kLa) in the presence of antifoam. Moreover, the antifoams did not affect the dissolved oxygen concentration of the cultures. A comparison of the amount of GFP retained in the cell by flow cytometry with that in the culture medium by fluorimetry suggested that addition of Antifoam A, Antifoam C or J673A increased the specific yield of GFP by increasing the proportion secreted into the medium.

Conclusions

We show that addition of a range of antifoaming agents to shake flask cultures ofP. pastorisincreases the total yield of the recombinant protein being produced. This is not only a simple method to increase the amount of protein in the culture, but our study also provides insight into how antifoams interact with microbial cell factories. Two mechanisms are apparent: one group of antifoams (Antifoam A, Antifoam C and J673A) increases the specific yield of GFP by increasing the total amount of protein produced and secreted per cell, whilst the second (P2000 or SB2121) increases the total yield by increasing the density of the culture.
The duration of diarrhea and fever is associated with growth faltering in rural Malawian children aged 6-18 months
Authors: Weisz Ariana , Meuli Gus , Thakwalakwa Chrissie , Trehan Indi , Maleta Kenneth , Manary Mark
Published: March 2011

Nutrition support programs that only focus upon better complementary feeding remain an insufficient means of limiting growth faltering in vulnerable populations of children. To determine if symptoms of acute infections correlate with the incidence of growth faltering in rural Malawian children, the associations between fever, diarrhea, and cough with anthropometric measures of stunting, wasting, and underweight were investigated. Data were analyzed from a trial where 209 children were provided with adequate complementary food and followed fortnightly from 6-18 months of age. Linear mixed model analysis was used to test for associations. Diarrheal disease was inversely associated with changes in height-for-age Z-score (HAZ), mid-upper arm circumference Z-score (MUACZ), and weight-for-age Z-score (WAZ). Fever was also inversely associated with changes in MUACZ and WAZ. These results suggest that initiatives to reduce febrile and diarrheal diseases are needed in conjunction with improved complementary feeding to limit growth faltering in rural Malawi.
Kriged and modeled ambient air levels of benzene in an urban environment: an exposure assessment study
Authors: Whitworth Kristina W , Symanski Elaine , Lai Dejian , Coker Ann L
Published: March 2011

Background

There is increasing concern regarding the potential adverse health effects of air pollution, particularly hazardous air pollutants (HAPs). However, quantifying exposure to these pollutants is problematic.

Objective

Our goal was to explore the utility of kriging, a spatial interpolation method, for exposure assessment in epidemiologic studies of HAPs. We used benzene as an example and compared census tract-level kriged predictions to estimates obtained from the 1999 U.S. EPA National Air Toxics Assessment (NATA), Assessment System for Population Exposure Nationwide (ASPEN) model.

Methods

Kriged predictions were generated for 649 census tracts in Harris County, Texas using estimates of annual benzene air concentrations from 17 monitoring sites operating in Harris and surrounding counties from 1998 to 2000. Year 1999 ASPEN modeled estimates were also obtained for each census tract. Spearman rank correlation analyses were performed on the modeled and kriged benzene levels. Weighted kappa statistics were computed to assess agreement between discretized kriged and modeled estimates of ambient air levels of benzene.

Results

There was modest correlation between the predicted and modeled values across census tracts. Overall, 56.2%, 40.7%, 31.5% and 28.2% of census tracts were classified as having 'low', 'medium-low', 'medium-high' and 'high' ambient air levels of benzene, respectively, comparing predicted and modeled benzene levels. The weighted kappa statistic was 0.26 (95% confidence interval (CI) = 0.20, 0.31), indicating poor agreement between the two methods.

Conclusions

There was a lack of concordance between predicted and modeled ambient air levels of benzene. Applying methods of spatial interpolation for assessing exposure to ambient air pollutants in health effect studies is hindered by the placement and number of existing stationary monitors collecting HAP data. Routine monitoring needs to be expanded if we are to use these data to better assess environmental health risks in the future.
Generation of novel cationic antimicrobial peptides from natural non-antimicrobial sequences by acid-amide substitution
Authors: Ueno Satoshi , Minaba Masaomi , Nishiuchi Yuji , Taichi Misako , Tamada Yasushi , Yamazaki Toshimasa , Kato Yusuke
Published: March 2011

Background

Cationic antimicrobial peptides (CAMPs) are well recognized to be promising as novel antimicrobial and antitumor agents. To obtain novel skeletons of CAMPs, we propose a simple strategy using acid-amide substitution (i.e. Glu→Gln, Asp→Asn) to confer net positive charge to natural non-antimicrobial sequences that have structures distinct from known CAMPs. The potential of this strategy was verified by a trial study.

Methods

The pro-regions of nematode cecropin P1-P3 (P1P-P3P) were selected as parent sequences. P1P-P3P and their acid-amide-substituted mutants (NP1P-NP3P) were chemically synthesized. Bactericidal and membrane-disruptive activities of these peptides were evaluated. Conformational changes were estimated from far-ultraviolet circular dichroism (CD) spectra.

Results

NP1P-NP3P acquired potent bactericidal activities via membrane-disruption although P1P-P3P were not antimicrobial. Far-ultraviolet CD spectra of NP1P-NP3P were similar to those of their parent peptides P1P-P3P, suggesting that NP1P-NP3P acquire microbicidal activity without remarkable conformational changes. NP1P-NP3P killed bacteria in almost parallel fashion with their membrane-disruptive activities, suggesting that the mode of action of those peptides was membrane-disruption. Interestingly, membrane-disruptive activity of NP1P-NP3P were highly diversified against acidic liposomes, indicating that the acid-amide-substituted nematode cecropin pro-region was expected to be a unique and promising skeleton for novel synthetic CAMPs with diversified membrane-discriminative properties.

Conclusions

The acid-amide substitution successfully generated some novel CAMPs in our trial study. These novel CAMPs were derived from natural non-antimicrobial sequences, and their sequences were completely distinct from any categories of known CAMPs, suggesting that such mutated natural sequences could be a promising source of novel skeletons of CAMPs.
Post - prandial rise of microvesicles in peripheral blood of healthy human donors
Authors: Šuštar Vid , Bedina-Zavec Apolonija , Štukelj Roman , Frank Mojca , Ogorevc Eva , Janša Rado , Mam Keriya , Veranič Peter , Kralj-Iglič Veronika
Published: March 2011

Background

Microvesicles isolated from body fluids are membrane - enclosed fragments of cell interior which carry information on the status of the organism. It is yet unclear how metabolism affects the number and composition of microvesicles in isolates from the peripheral blood.

Aim

To study the post - prandial effect on microvesicles in isolates from the peripheral blood of 21 healthy donors, in relation to blood cholesterol and blood glucose concentrations.

Results

The average number of microvesicles in the isolates increased 5 hours post - prandially by 52%; the increase was statistically significant (p = 0.01) with the power P = 0.68, while the average total blood cholesterol concentration, average low density lipoprotein cholesterol concentration (LDL-C) and average high density lipoprotein cholesterol concentration (HDL-C) all remained within 2% of their fasting values. We found an 11% increase in triglycerides (p = 0.12) and a 6% decrease in blood glucose (p < 0.01, P = 0.74). The post - prandial number of microvesicles negatively correlated with the post - fasting total cholesterol concentration (r = - 0.46, p = 0.035) while the difference in the number of microvesicles in the isolates between post - prandial and post - fasting states negatively correlated with the respective difference in blood glucose concentration (r = - 0.39, p = 0.05).

Conclusions

In a population of healthy human subjects the number of microvesicles in isolates from peripheral blood increased in the post - prandial state. The increase in the number of microvesicles was affected by the fasting concentration of cholesterol and correlated with the decrease in blood glucose.
Proteomics: a subcellular look at spermatozoa
Authors: du Plessis Stefan S , Kashou Anthony H , Benjamin David J , Yadav Satya P , Agarwal Ashok
Published: March 2011

Background

Male-factor infertility presents a vexing problem for many reproductively active couples. Many studies have focused on abnormal sperm parameters. Recent advances in proteomic techniques, especially in mass spectrometry, have aided in the study of sperm and more specifically, sperm proteins. The aim of this study was to review the current literature on the various proteomic techniques, and their usefulness in diagnosing sperm dysfunction and potential applications in the clinical setting.

Methods

Review of PubMed database. Key words: spermatozoa, proteomics, protein, proteome, 2D-PAGE, mass spectrometry.

Results

Recently employed proteomic methods, such as two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in gel electrophoresis, have identified numerous sperm-specific proteins. They also have provided a further understanding of protein function involved in sperm processes and for the differentiation between normal and abnormal states. In addition, studies on the sperm proteome have demonstrated the importance of post-translational modifications, and their ability to bring about physiological changes in sperm function. No longer do researchers believe that in order for them to elucidate the biochemical functions of genes, mere knowledge of the human genome sequence is sufficient. Moreover, a greater understanding of the physiological function of every protein in the tissue-specific proteome is essential in order to unravel the biological display of the human genome.

Conclusion

Recent advances in proteomic techniques have provided insight into sperm function and dysfunction. Several multidimensional separation techniques can be utilized to identify and characterize spermatozoa. Future developments in bioinformatics can further assist researchers in understanding the vast amount of data collected in proteomic studies. Moreover, such advances in proteomics may help to decipher metabolites which can act as biomarkers in the detection of sperm impairments and to potentially develop treatment for infertile couples.

Further comprehensive studies on sperm-specific proteome, mechanisms of protein function and its proteolytic regulation, biomarkers and functional pathways, such as oxidative-stress induced mechanisms, will provide better insight into physiological functions of the spermatozoa. Large-scale proteomic studies using purified protein assays will eventually lead to the development of novel biomarkers that may allow for detection of disease states, genetic abnormalities, and risk factors for male infertility. Ultimately, these biomarkers will allow for a better diagnosis of sperm dysfunction and aid in drug development.
Double suicide genes driven by kinase domain insert containing receptor promoter selectively kill human lung cancer cells
Authors: Ma Junrong , Li Mi , Mei Longyong , Zhou Qinghua , Liu Lunxu , Yu Xijie , Che Guowei
Published: March 2011

Background

To investigate the selective killing efficacy of the double suicide genes driven by KDR promoter.

Materials and methods

A double suicide gene system with the KDR promoter, pcDNA3-KDRp-CDglyTK, was constructed and transfected into lung cancer cell lines L9981 and NL9980, and human hepatocellular carcinoma cell line HepG2. The efficiency and specificity of the double suicide gene system were assayed by in vitro cellular proliferation and apoptosis, as well as in vivo xenograft studies.

Results

The transgenic CD and TK genes were only expressed in L9981 and NL9980 but not in HepG2 cells. Pre-treating transfected cells with 5-Fc and GCV significantly reduced proliferation, enhanced apoptosis in L9981 and NL9980 but not in HepG2 cells. The tumor formed by L9981 and NL9980 cells with the double suicide gene system was much smaller in vivo.

Conclusion

Tumor targeted expression of CDglyTK gene driven by KDR promotor represents a novel strategy for effective gene therapy of tumor with intrinsic KDR.