Progesterone, glucocorticoid, but not estrogen receptor mRNA is altered in breast cancer stroma

Our laboratory has previously found that anti-mitogenic nuclear receptor mRNA is elevated in late stage tumours and this study was performed to scrutinize the possibility of cancer-stroma crosstalk using hormone signaling in these tissues. RNA levels in stromal tissue were examined for the estrogen α, estrogen β, androgen, progesterone and glucocorticoid nuclear receptors by a semi-quantitative PCR. Significant differences in expression between the cancer stroma and control tissue were seen, analyzing for both cancer grade and estrogen receptor status. Stroma and control tissue were significantly different for the progesterone and glucocorticoid nuclear receptors (p = 5.908 × 10−7 and 2.761 × 10−5, respectively). Glucocorticoid receptor also showed a significant increase to mRNA levels in the stroma of estrogen receptor negative tumours (p = 5.85 × 10−5). By contrast, the estrogen receptors α and β, those most closely associated with breast tissue growth, showed no significant change in mRNA (p = 0.372 and 0.655, respectively). Androgen receptor mRNA also remained unaffected (p = 0.174).

The estrogen receptor 1 G594A polymorphism is associated with migraine susceptibility in two independent case/control groups

Migraine is a painful and debilitating disorder with a significant genetic component. Steroid hormones, in particular estrogen, have long been considered to play a role in migraine, as variations in hormone levels are associated with migraine onset in many sufferers of the disorder. Steroid hormones mediate their activity via hormone receptors, which have a wide tissue distribution. Estrogen receptors have been localized to the brain in regions considered to be involved in migraine pathogenesis. Hence it is possible that genetic variation in the estrogen receptor gene may play a role in migraine susceptibility. This study thus examined the estrogen receptor 1 (ESRα) gene for a potential role in migraine pathogenesis and susceptibility. A population-based cohort of 224 migraine sufferers and 224 matched controls were genotyped for the G594A polymorphism located in exon 8 of the ESR1 gene. Statistical analysis indicated a significant difference between migraineurs and non-migraineurs in both the allele frequencies (P=0.003) and genotype distributions (P=0.008) in this sample. An independent follow-up study was then undertaken using this marker in an additional population-based cohort of 260 migraine sufferers and 260 matched controls. This resulted in a significant association between the two groups with regard to allele frequencies (P=8×10−6) and genotype distributions (P=4×10−5). Our findings support the hypothesis that genetic variation in hormone receptors, in particular the ESR1 gene, may play a role in migraine.

β-Actin—an unsuitable internal control for RT-PCR

Despite reports confirming cell-cycle dependent gene expression and a number of studies describing specific circumstances in which β-actin is also regulated, the mRNA for β-actin remains a widely used housekeeping gene internal control. Utilizing differential reverse transcriptase-polymerase chain reaction (RT-PCR), we report here the dose-dependent inhibition of β-actin by matrigel. This was detected by comparison to the very moderate inhibition of the target gene, membrane type-1 matrix metalloproteinase (MT1-MMP), with results independently confirmed by similar findings on MT1-MMP expression using competitive RT-PCR. Furthermore, RT-PCR of the housekeeping gene 18 Svedberg Units (S) rRNA demonstrated excellent consistency, reproducibility and non-regulation by a matrigel treatment. We conclude that β-actin is highly regulated by matrigel and therefore unsuitable as an internal control in this treatment. Hence, these findings suggest that researchers have a responsibility to ensure that the housekeeping gene of choice is not regulated in their specific application, as such regulation may dramatically affect the accuracy of their results. This study reinforces the necessity for minimally regulated housekeeping genes such as 18S rRNA, and the superiority of competitive templates as internal controls for quantitative applications of RT-PCR.

Association of estrogen receptor and glucocorticoid receptor gene polymorphisms with sporadic breast cancer

We have utilized a cross-sectional association approach to investigate sporadic breast cancer. Polymorphisms in 2 candidate genes, ESRalpha and GRL, were examined in an unrelated breast cancer-affected and age-matched control population. Several polymorphic regions within the ESRalpha gene have been identified, and some alleles of these polymorphisms have been found to occur at increased levels in breast-cancer patients. Additionally, variations in GRL have the potential to disrupt cell transcription and may be associated with cancer formation. We analyzed 3 polymorphisms, from codons 10 (TCT to TCC), 325 (CCC to CCG) and 594 (ACA to ACG) of ESRalpha, and a highly polymorphic dinucleotide repeat, D5S207, located within 200 kb of the GRL. When allelic frequencies of the codon 594 (exon 8) ESR polymorphism were compared between affected and unaffected populations, a significant difference was observed (p = 0.005). Results from the D5S207 dinucleotide repeat located near GRL also indicated a significant difference between the tested case and control populations (p = 0.001). Allelic frequencies of the codon 10 and codon 325 ESR polymorphisms were not significantly different between populations (p = 0.152 and 0.181, respectively). Our results indicate that specific alleles of the ESR gene (alpha subtype) and a marker for the GRL gene locus are associated with sporadic breast-cancer development in the tested Caucasian population and justify further investigation of the role of these and other nuclear steroid receptors in the etiology of breast cancer.

Excursion guidance criteria to guide control of peak emission and exposure to airborne engineered particles

The overall aim of our research was to characterize airborne particles from selected nanotechnology processes and to utilize the data to develop and test quantitative particle concentration-based criteria that can be used to trigger an assessment of particle emission controls. We investigated particle number concentration (PNC), particle mass (PM) concentration, count median diameter (CMD), alveolar deposited surface area, elemental composition, and morphology from sampling of aerosols arising from six nanotechnology processes. These included fibrous and non-fibrous particles, including carbon nanotubes (CNTs). We adopted standard occupational hygiene principles in relation to controlling peak emission and exposures, as outlined by both Safe Work Australia, (1) and the American Conference of Governmental Industrial Hygienists (ACGIH®). (2) The results from the study were used to analyses peak and 30-minute averaged particle number and mass concentration values measured during the operation of the nanotechnology processes. Analysis of peak (highest value recorded) and 30-minute averaged particle number and mass concentration values revealed: Peak PNC20–1000 nm emitted from the nanotechnology processes were up to three orders of magnitude greater than the local background particle concentration (LBPC). Peak PNC300–3000 nm was up to an order of magnitude greater, and PM2.5 concentrations up to four orders of magnitude greater. For three of these nanotechnology processes, the 30-minute average particle number and mass concentrations were also significantly different from the LBPC (p-value < 0.001). We propose emission or exposure controls may need to be implemented or modified, or further assessment of the controls be undertaken, if concentrations exceed three times the LBPC, which is also used as the local particle reference value, for more than a total of 30 minutes during a workday, and/or if a single short-term measurement exceeds five times the local particle reference value. The use of these quantitative criteria, which we are terming the universal excursion guidance criteria, will account for the typical variation in LBPC and inaccuracy of instruments, while precautionary enough to highlight peaks in particle concentration likely to be associated with particle emission from the nanotechnology process. Recommendations on when to utilize local excursion guidance criteria are also provided.

Out of control : Emergent cultural landscapes and political change in urban Vietnam

This paper plots the recent changes in the uses of public space in Hanoi, Vietnam. It is argued that the economic and social changes in contemporary Vietnam have paved the way for a dramatic transformation in the ways in which streets, pavements and markets are experienced and imagined by the populace. The efflorescence of individual mobility, street-trading and public crowding around certain popular events has led to the emergence of a distinct public sphere, one which is not immune from state control and censure but which is a flagrant rebuttal of the state's appeal. The immediate struggles over space herald a new discursive arena for the contest over Vietnamese national imagery as represented in cultural heritage and public space, memorials and state-controlled events which the public are rapidly deserting. The paper concludes by suggesting that the everyday cultural practices that have created a bustling streetlife in urban Vietnam will inevitably provide the vitality and spectacle for the destabilisation of state control in a struggle for meanings in public space.

Non-linear position control for hover and automatic landing of unmanned aerial vehicles

This study presents a disturbance attenuation controller for horizontal position stabilisation for hover and automatic landings of a rotary-wing unmanned aerial vehicle (RUAV) operating close to the landing deck in rough seas. Based on a helicopter model representing aerodynamics during the landing phase, a non-linear state feedback H∞ controller is designed to achieve rapid horizontal position tracking in a gusty environment. Practical constraints including flapping dynamics, servo dynamics and time lag effect are considered. A high-fidelity closed-loop simulation using parameters of the Vario XLC gas-turbine helicopter verifies performance of the proposed horizontal position controller. The proposed controller not only increases the disturbance attenuation capability of the RUAV, but also enables rapid position response when gusts occur. Comparative studies show that the H∞ controller exhibits performance improvement and can be applied to ship/RUAV landing systems.

Load frequency control in a microgrid : challenges and improvements

A microgrid can span over a large area, especially in rural townships. In such cases, the distributed generators (DGs) must be controlled in a decentralized fashion, based on the locally available measurements. The main concerns are control of system voltage magnitude and frequency, which can either lead to system instability or voltage collapse. In this chapter, the operational challenges of load frequency control in a microgrid are discussed and few methods are proposed to meet these challenges. In particular, issues of power sharing, power quality and system stability are addressed, when the system operates under decentralized control. The main focus of this chapter is to provide solutions to improve the system performance in different situations. The scenarios considered are (a) when the system stability margin is low, (b) when the line impedance has a high R to X ratio, (c) when the system contains unbalanced and/or distorted loads. Also a scheme is proposed in which a microgrid can be frequency isolated from a utility grid while being capable of bidirectional power transfer. In all these cases, the use of angle droop in converter interfaced DGs is adopted. It has been shown that this results in a more responsive control action compared to the traditional frequency based droop control.

Analysis of Sdhd and Mmp12 in an affected solar keratosis and control cohort

The incidence of Squamous Cell Carcinoma (SCG) is growing in certain populations to the extent that it is now the most common skin lesion in young men and women in high ultraviolet exposure regions such as Queensland. In terms of incidence up to 40% of the Australian population over 40 years of age is thought to possess the precancerous Solar Keratosis (SK) lesion and with a small, but significant, chance of progression into SCC, understanding the genetic events that play a role in this process is essential. The major aims of this study were to analyse whole blood derived samples for DNA aberrations in genes associated with tumour development and cellular maintenance, with the ultimate aim of identifying genes associated with non-melanoma skin cancer development. More specifically the first aim of this project was to analyse the SDHD and MMP12 genes via Dual-Labelled Probe Real-Time PCR for copy number aberrations in an affected Solar Keratosis and control cohort. It was found that 12 samples had identifiable copy-number aberrations in either the SDHD or MMP12 gene (this means that a genetic section of either of these two genes is aberrantly amplified or deleted), with five of the samples exhibiting aberrations in both genes. The significance of this study is the contribution to the knowledge of the genetic pathways that are malformed in the progression and development of the pre-cancerous skin lesion Solar Keratosis. © 2008 Springer Science+Business Media, LLC.

Detection of mRNA for nuclear receptor co-activators 1 and 3 in archival breast cancer tissue and surrounding stroma : a tissue expression study

Before the age of 75 years, approximately 10% of women will be diagnosed with breast cancer, one of the most common malignancies and a leading cause of death among women. The objective of this study was to determine if expression of the nuclear receptor coactivators 1 and 3 (NCoA1 and NCoA3) varied in breast cancer grades. RNA was extracted from 25 breast tumours and transcribed into cDNA which underwent semi-quantitative polymerase chain reaction, normalised using 18S. Analysis indicated that an expression change for NCoA1 in cancer grades and estrogen receptor alpha negative tissue (P= 0.028 and 0.001 respectively). NCoA1 expression increased in grade 3 and estrogen receptor alpha negative tumours, compared to controls. NCoA3 showed a similar, but not significant, trend in grade and a non-significant decrease in estrogen receptor alpha negative tissues. Expression of NCoA1 in late stage and estrogen receptor alpha negative breast tumours may have implications to breast cancer treatment, particularly in the area of manipulation of hormone signalling systems in advanced tumours.

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.

GEF-H1-RhoA signaling pathway mediates LPS-induced NF-κB transactivation and IL-8 synthesis in endothelial cells

Secretion of proinflammatory cytokines by LPS activated endothelial cells contributes substantially to the pathogenesis of sepsis. However, the mechanism involved in this process is not well understood. In the present study, we determined the roles of GEF-H1 (Guanine-nucleotide exchange factor-H1)-RhoA signalling in LPS-induced interleukin-8 (IL-8, CXCL8) production in endothelial cells. First, we observed that GEF-H1 expression was upregulated in a dose- and time-dependent manner as consistent with TLR4 (Toll-like receptor 4) expression after LPS stimulation. Afterwards, Clostridium difficile toxin B-10463 (TcdB-10463), an inhibitor of Rho activities, reduced LPS-induced NF-κB phosphorylation. Inhibition of GEF-H1 and RhoA expression reduced LPS-induced NF-κB and p38 phosphorylation. TLR4 knockout blocked LPS-induced activity of RhoA, however, MyD88 knockout did not impair the LPS-induced activity of RhoA. Nevertheless, TLR4 and MyD88 knockout both significantly inhibited transactivation of NF-κB. GEF-H1-RhoA and MyD88 both induced significant changes in NF-κB transactivation and IL-8 synthesis. Co-inhibition of GEF-H1-RhoA and p38 expression produced similar inhibitory effects on LPS-induced NF-κB transactivation and IL-8 synthesis as inhibition of p38 expression alone, thus confirming that activation of p38 was essential for the GEF-H1-RhoA signalling pathway to induce NF-κB transactivation and IL-8 synthesis. Taken together, these results demonstrate that LPS-induced NF-κB activation and IL-8 synthesis in endothelial cells are regulated by the MyD88 pathway and GEF-H1-RhoA pathway.

Postprandial total and HMW adiponectin following a high-fat meal in lean, obese and diabetic men

BACKGROUND/OBJECTIVES: Recent work suggests that macronutrients are pro-inflammatory and promote oxidative stress. Reports of postprandial regulation of total adiponectin have been mixed, and there is limited information regarding postprandial changes in high molecular weight (HMW) adiponectin. The aim of this study was to assess the effect of a standardised high-fat meal on metabolic variables, adiponectin (total and HMW), and markers of inflammation and oxidative stress in: (i) lean, (ii) obese non-diabetic and (iii) men with type 2 diabetes mellitus (T2DM). SUBJECTS/METHODS: Male subjects: lean (n=10), obese (n=10) and T2DM (n=10) were studied for 6 h following both a high-fat meal and water control. Metabolic variables (glucose, insulin, triglycerides), inflammatory markers (interleukin-6 (IL6), tumour necrosis factor (TNF)α, high-sensitivity C-reactive protein (hsCRP), nuclear factor (NF)κB expression in peripheral blood mononuclear cells (p65)), indicators of oxidative stress (oxidised low density lipoprotein (oxLDL), protein carbonyl) and adiponectin (total and HMW) were measured. RESULTS: No significant changes in TNFα, p65, oxLDL or protein carbonyl concentrations were observed. Overall, postprandial IL6 decreased in subjects with T2DM but increased in lean subjects, whereas hsCRP decreased in the lean cohort and increased in obese subjects. There was no overall postprandial change in total or HMW adiponectin in any group. Total adiponectin concentrations changed over time following the water control, and the response was significantly different in lean subjects compared with subjects with T2DM (P=0.04). CONCLUSIONS: No consistent significant postprandial inflammation, oxidative stress or regulation of adiponectin was observed in this study. Findings from the water control suggest differential basal regulation of total adiponectin in T2DM compared with lean controls.

Control, shift, insert : living and enacting policy in teachers' day/night work

This thesis investigates the experiences of teachers who trialled an electronic curriculum and assessment tool in the wider context of text-mediated ruling relations organising their work. Problematised as policy and text, this tool is interrogated as a 'solution' to problems perceived in teachers' work in an era of increased accountability. It provides evidence that teachers' work is shaped by forces operating outside their control and mediated by the policy discourses and subjectivities available to them.

Hysteretic multilevel NPC converter control for a Switch Mode Assisted Linear Amplifier

A Switch-Mode Assisted Linear Amplifier (SMALA) combines the high quality of a linear amplifier required for audio applications with the high efficiency of a switch-mode amplifier. The careful choice of current sense point and switch placement allows a simple non-isolated hysteresis current controller for the switch-mode section. This paper explains the extension of the hysteresis current controller for the control of a three level Neutral Point Clamped (NPC) converter, with simulations as proof of concept. The NPC topology allows the use of lower voltage switches and lower switching frequencies to implement high power audio amplifiers using the SMALA topology.