Differential gene expression in breast cancer cell lines and stroma-tumor differences in microdissected breast cancer biopsies revealed by display array analysis

To examine gene-expression patterning in late-stage breast cancer biopsies, we used a microdissection technique to separate tumor from the surrounding breast tissue or stroma. A DD-PCR protocol was then used to amplify expressed products, which were resolved using PAGE and used as probe to hybridize with representative human arrays and cDNA libraries. The probe derived from the tumor–stroma comparison was hybridized with a gene array and an arrayed cDNA library derived from a GCT of bone; 21 known genes or expressed sequence tags were detected, of which 17 showed differential expression. These included factors associated with epithelial to mesenchymal transition (vimentin), the cargo selection protein (TIP47) and the signal transducer and activator of transcription (STAT3). Northern blot analysis was used to confirm those genes also expressed by representative breast cancer cell lines. Notably, 6 genes of unknown function were restricted to tumor while the majority of stroma-associated genes were known. When applied to transformed breast cancer cell lines (MDA-MB-435 and T47D) that are known to have different metastatic potential, DD array analysis revealed a further 20 genes; 17 of these genes showed differential expression. Use of microdissection and the DD-PCR array protocol allowed us to identify factors whose localized expression within the breast may play a role in abnormal breast development or breast carcinogenesis.

Composites for delivery of therapeutics : combining melt electrospun scaffolds with loaded electrosprayed microparticles

A novel strategy is reported to produce biodegradable microfiber-scaffolds layered with high densities of microparticles encapsulating a model protein. Direct electrospraying on highly porous melt electrospun scaffolds provides a reproducible scaffold coating throughout the entire architecture. The burst release of protein is significantly reduced due to the immobilization of microparticles on the surface of the scaffold and release mechanisms are dependent on protein-polymer interactions. The composite scaffolds have a positive biological effect in contact with precursor osteoblast cells up to 18 days in culture. The scaffold design achieved with the techniques presented here endorses these new composite scaffolds as promising templates for growth factor delivery.

Expression of glucocorticoid and progesterone nuclear receptor genes in archival breast cancer tissue.

BACKGROUND: Previous studies in our laboratory have shown associations of specific nuclear receptor gene variants with sporadic breast cancer. In order to investigate these findings further, we conducted the present study to determine whether expression levels of the progesterone and glucocorticoid nuclear receptor genes vary in different breast cancer grades. METHODS: RNA was extracted from paraffin-embedded archival breast tumour tissue and converted into cDNA. Sample cDNA underwent PCR using labelled primers to enable quantitation of mRNA expression. Expression data were normalized against the 18S ribosomal gene multiplex and analyzed using analysis of variance. RESULTS: Analysis of variance indicated a variable level of expression of both genes with regard to breast cancer grade (P = 0.00033 for glucocorticoid receptor and P = 0.023 for progesterone receptor). CONCLUSION: Statistical analysis indicated that expression of the progesterone nuclear receptor is elevated in late grade breast cancer tissue.

Spectroscopic correlation analysis of NMR-based metabonomics in exercise science

Spectroscopic studies of complex clinical fluids have led to the application of a more holistic approach to their chemical analysis becoming more popular and widely employed. The efficient and effective interpretation of multidimensional spectroscopic data relies on many chemometric techniques and one such group of tools is represented by so-called correlation analysis methods. Typical of these techniques are two-dimensional correlation analysis and statistical total correlation spectroscopy (STOCSY). Whilst the former has largely been applied to optical spectroscopic analysis, STOCSY was developed and has been applied almost exclusively to NMR metabonomic studies. Using a 1H NMR study of human blood plasma, from subjects recovering from exhaustive exercise trials, the basic concepts and applications of these techniques are examined. Typical information from their application to NMR-based metabonomics is presented and their value in aiding interpretation of NMR data obtained from biological systems is illustrated. Major energy metabolites are identified in the NMR spectra and the dynamics of their appearance and removal from plasma during exercise recovery are illustrated and discussed. The complementary nature of two-dimensional correlation analysis and statistical total correlation spectroscopy are highlighted.

Analysis on causes of death of rural residents in Shouguang City [寿光市农村人群慢性病死因分析]

Among people living in rural areas of Shouguang City, Shandong, China, 88% of deaths were caused by chronic non-communicable diseases. Cardiovascular diseases, respiratory diseases and cancers accounted for 97% of all chronic disease causes and 85% of all causes of death. The proportion of chronic causes increased by 27% from 1993 to 2000. However, the mortality of respiratory diseases showed a decreasing trend over time. Abstract in Chinese 篇首： 随着生活水平的提高,慢性非传染性疾病在居民死因谱中所占比重越来越大,为准确反映居民的死因状况,为农村地区慢性病的控制工作提供科学依据,本文对寿光市农村疾病监测点1993～2000年居民的慢性病死因进行了分析.

Herceptest: HER2 expression and gene amplification in non-small cell lung cancer

HER2 is an erbB/HER type I tyrosine kinase receptor that is frequently over-expressed in malignant epithelial tumours. Herceptin, a humanised mouse monoclonal antibody to HER2, is proven therapeutically in the management of metastatic breast cancer, significantly prolonging survival when combined with cytotoxic chemotherapeutic agents. Immunohistochemical studies suggest that non-small-cell lung cancer (NSCLC) tumours may over-express HER2. Our aim was to evaluate HER2 gene amplification and semi-quantitative immuno-expression in NSCLC. A total of 344 NSCLC cases were immunostained for HER2 expression in 2 centres using the HercepTest. Fluorescence in situ hybridisation (FISH) analysis for HER2 gene amplification was performed on most positive cases and a subset of negative cases. Fifteen cases (4.3%) demonstrated 2+ or 3+ membranous HER2 immuno-expression. There was no correlation between immuno-expression and tumour histology or grade. Tumours from higher-stage disease were more often HercepTest-positive (p < 0.001). All 4 HercepTest 3 + cases demonstrated gene amplification. One of the 5 2+ cases tested for gene amplification showed areas of borderline amplification and areas of polyploidy. None of the 19 HercepTest-negative cases demonstrated gene amplification or polyploidy (p < 0.001). Gene amplification was demonstrated in all HercepTest 3+ scoring NSCLC cases. Unlike breast cancer, gene amplification and HER2 protein over-expression assessed by the HercepTest appeared to be uncommon in NSCLC. Herceptin may therefore target only a small proportion of NSCLC tumours and be of limited clinical value in this disease, particularly in the adjuvant setting. © 2001 Wiley-Liss, Inc.

Prognostic factors in patients with advanced non-small cell lung cancer : data from the phase III FLEX study

The FLEX study demonstrated that the addition of cetuximab to chemotherapy significantly improved overall survival in the first-line treatment of patients with advanced non-small cell lung cancer (NSCLC). In the FLEX intention to treat (ITT) population, we investigated the prognostic significance of particular baseline characteristics. Individual patient data from the treatment arms of the ITT population of the FLEX study were combined. Univariable and multivariable Cox regression models were used to investigate variables with potential prognostic value. The ITT population comprised 1125 patients. In the univariable analysis, longer median survival times were apparent for females compared with males (12.7 vs 9.3 months); patients with an Eastern Cooperative Oncology Group performance status (ECOG PS) of 0 compared with 1 compared with 2 (13.5 vs 10.6 vs 5.9 months); never smokers compared with former smokers compared with current smokers (14.6 vs 11.1 vs 9.0); Asians compared with Caucasians (19.5 vs 9.6 months); patients with adenocarcinoma compared with squamous cell carcinoma (12.4 vs 9.3 months) and those with metastases to one site compared with two sites compared with three or more sites (12.4 months vs 9.8 months vs 6.4 months). Age (<65 vs ≥65 years), tumor stage (IIIB with pleural effusion vs IV) and percentage of tumor cells expressing EGFR (<40% vs ≥40%) were not identified as possible prognostic factors in relation to survival time. In multivariable analysis, a stepwise selection procedure identified age (<65 vs ≥65 years), gender, ECOG PS, smoking status, region, tumor histology, and number of organs involved as independent factors of prognostic value. In summary, in patients with advanced NSCLC enrolled in the FLEX study, and consistent with previous analyses, particular patient and disease characteristics at baseline were shown to be independent factors of prognostic value. The FLEX study is registered with ClinicalTrials.gov, number NCT00148798. © 2012 Elsevier Ireland Ltd.

High-throughput vectors for efficient gene silencing in plants

A major challenge in the post-genome era of plant biology is to determine the functions of all genes in the plant genome. A straightforward approach to this problem is to reduce or knockout expression of a gene with the hope of seeing a phenotype that is suggestive of its function. Insertional mutagenesis is a useful tool for this type of study but is limited by gene redundancy, lethal knockouts, non-tagged mutants, and the inability to target the inserted element to a specific gene. The efficacy of gene silencing in plants using inverted-repeat transgene constructs that encode a hairpin RNA (hpRNA) has been demonstrated by a number of groups, and has several advantages over insertional mutagenesis. In this paper we describe two improved pHellsgate vectors that facilitate rapid generation of hpRNA-encoding constructs, pHellsgate 4 allows the production of an hpRNA construct in a single step from a single polymerase chain reaction product, while pHellsgate 8 requires a two-step process via an intermediate vector. We show that these vectors are effective at silencing three endogenous genes in Arabidopsis, FLOWERING LOCUS C, PHYTOENE DESATURASE and ETHYLENE INSENSITIVE 2. We also show that a construct of sequences from two genes silences both genes.

CD151 is associated with prostate cancer cell invasion and lymphangiogenesis in vivo

CD151, a member of the tetraspanin family, is associated with regulation of migration of normal and tumour cells via cell surface microdomain formation. CD151 was found in our laboratory to have a prognostic value in prostate cancer and is a promoter of prostate cancer migration and invasion. These roles involve association with integrins on both cell-cell and cell-stroma levels. Furthermore, CD151 plays a role in endothelial cell motility. CD151 expression was examined in three commonly used prostate cancer cell lines. We investigated CD151 expression, angiogenesis (microvessel density; MVD) and lymphangiogenesis (lymphatic vessel density; LVD) in an orthotopic xenograft model of prostate cancer in matched tumours from primary and secondary sites. CD151 was found to be heterogeneously expressed across different prostate cancer cell lines and the levels of CD151 expression were significantly higher in the highly tumorigenic, androgen-insensitive cells PC-3 and DU-145 compared to the androgen-sensitive cell line LNCaP (P<0.05). The majority of in vivo xenografts developed pelvic lymph node metastases. Importantly, primary tumours that developed metastasis had significantly higher CD151 expression and MVD compared to those which did not develop metastasis (P<0.05). We identified, for the first time, that CD151 expression is associated with LVD in prostate cancer. These findings underscore the potential role of CD151 and angiogenesis in the metastatic potential of prostate cancer. CD151 has a prognostic value in this mouse model of prostate cancer and may play a role in lymphangiogenesis. CD151 is likely an important regulator of cancer cell communication with the surrounding microenvironment.

Spiritual wisdom of Taoism in business : through the lens of interpretation realism in a Cisco end-to-end case study

This paper explores how the amalgamated wisdom of East and West can instigate a wisdombased renaissance of humanistic epistemology (Rooney & McKenna, 2005) to provide a platform of harmony in managing knowledge-worker productivity, one of the biggest management challenges of the 21st century (Drucker, 1999). The paper invites further discussions from the social and business research communities on the significance of "interpretation realism" technique in comprehending philosophies of Lao Tzu Confucius and Sun Tzu (Lao/Confucius/Sun] written in "Classical Chinese." This paper concludes with a call to build prudent, responsible practices in management which affects the daily lives of many (Rooney & McKenna, 2005) in today's knowledgebased economy. Interpretation Realism will be applied to an analysis of three Chinese classics of Lao/Confucius/Sun which have been embodied in the Chinese culture for over 2,500 years. Comprehending Lao/Confucius/Sun's philosophies is the first step towards understanding Classical Chinese culture. However, interpreting Chinese subtlety in language and the yin and yang circular synthesis in their mode of thinking is very different to understanding Western thought with its open communication and its linear, analytical pattern of Aristotelian/Platonic wisdom (Zuo, 2012). Furthermore, Eastern ways of communication are relatively indirect and mediatory in culture. Western ways of communication are relatively direct and litigious in culture (Goh, 2002). Furthermore, Lao/Confucius/Sun's philosophies are difficult to comprehend as there are four written Chinese formats and over 250 dialects: Pre-classical Chinese Classical Chinese Literary Chinese and modern Vernacular Chinese Because Classical Chinese is poetic, comprehension requires a mixed approach of interpretation realism combining logical reasoning behind "word splitting word occurrences", "empathetic metaphor" and "poetic appreciation of word.

Projecting future transmission of malaria under climate change scenarios : challenges and research needs

There has been an intense debate about climatic impacts on the transmission of malaria. It is vitally important to accurately project future impacts of climate change on malaria to support effective policy–making and intervention activity concerning malaria control and prevention. This paper critically reviewed the published literature and examined both key findings and methodological issues in projecting future impacts of climate change on malaria transmission. A literature search was conducted using the electronic databases MEDLINE, Web of Science and PubMed. The projected impacts of climate change on malaria transmission were spatially heterogeneous and somewhat inconsistent. The variation in results may be explained by the interaction of climatic factors and malaria transmission cycles, variations in projection frameworks and uncertainties of future socioecological (including climate) changes. Current knowledge gaps are identified, future research directions are proposed and public health implications are assessed. Improving the understanding of the dynamic effects of climate on malaria transmission cycles, the advancement of modelling techniques and the incorporation of uncertainties in future socioecological changes are critical factors for projecting the impact of climate change on malaria transmission.

Critical reflections on the use of participatory methodologies to build evaluation capacities in international development organisations

"In this chapter the authors present a critique of Participatory Evaluation as worked in development projects, in this case, in Nepal. The article works between established claims that Participatory Evaluation builds capacity at programmatic and organisational levels, and the specific experiences of these claims in the authors’ current work. They highlight the need to address key difficulties such as high turn-over of staff and resulting loss of capacity to engage in Participatory Evaluation, and the difficulty of communication between academic as compared with local practical wisdoms. A key issue is the challenge of addressing the inevitable issues of power inequities that such approaches encounter. While Participatory Evaluation has been around for some time, it has only enjoyed more widespread recognition of its value in comparatively recent times, with its uptake in international development environments. To this extent, the practice is still in its early stages of development, and Jo, June and Michael’s work contributes to strengthening and more comprehensively understanding it. With regard to the meta-theme of this publication, this chapter is an example of how context not only influences the methodology to be used and the praxis of how it is to be used, but contributes to early explication of the core nature of an emerging methodology."

Women's health in the global village

As we stand at the beginning of the 21st century and behold the world before us, it seems that we are living in a time of profound change. Everywhere we look change seems afoot, demolishing our traditional securities and hastily building new ones in their place. Modern medical science has been an integral part of this change. It is not possible to ignore the advances of modern medicine nor the realities of scientific uncertainties for they are part of the shared context of our lives today. I In the past 50 years we have witnessed the discovery of DNA and more recently the mapping of the human genome, the birth of the world's first in-vitro fertilisation baby, followed by thousands worldwide in the period since, the discovery of human stem cells and the birth of Dolly the cloned sheep in Scotland. Furthermore, the processes of globalisation have ensured that an event that occurs on one side of the globe becomes an item on the evening news on the other side, creating the impression that all change takes place on our doorstep. Some of these events have provoked deep angst in the community, sparking public debate over the ethics of science and the boundaries to be imposed by law. All of these developments have changed the realm of the possible. While these advances in medical science spark debate in the developed countries, in less developed countries high rates of infectious diseases and infant and maternal mortality and the challenges of access to adequate food and clean water are priorities, highlighting international differences in health care. This article explores these differences through an analysis of globalisation and reproduction. It seeks to analyse both the meaning of globalisation and the impact of globalising trends on health laws and policies as regulators of women's health within the global village.