The prognostic utility of the transcription factor SRF in docetaxel-resistant prostate cancer: in-vitro discovery and in-vivo validation.

BACKGROUND: Docetaxel based therapy is one of the first line chemotherapeutic agents for the treatment of metastatic castrate-resistant prostate cancer. However, one of the major obstacles in the treatment of these patients is docetaxel-resistance. Defining the mechanisms of resistance so as to inform subsequent treatment options and combinations represents a challenge for clinicians and scientists. Previous work by our group has shown complex changes in pro and anti-apoptotic proteins in the development of resistance to docetaxel. Targeting these changes individually does not significantly impact on the resistant phenotype but understanding the central signalling pathways and transcription factors (TFs) which control these could represent a more appropriate therapeutic targeting approach. METHODS: Using a number of docetaxel-resistant sublines of PC-3 cells, we have undertaken a transcriptomic analysis by expression microarray using the Affymetrix Human Gene 1.0 ST Array and in conjunction with bioinformatic analyses undertook to predict dysregulated TFs in docetaxel resistant prostate cancer. The clinical significance of this prediction was ascertained by performing immunohistochemical (IHC) analysis of an identified TF (SRF) in the metastatic sites from men who died of advanced CRPC. Investigation of the functional role of SRF was examined by manipulating SRF using SiRNA in a docetaxel-resistant PC-3 cell line model. RESULTS: The transcription factors identified include serum response factor (SRF), nuclear factor kappa-B (NFκB), heat shock factor protein 1 (HSF1), testicular receptor 2 & 4 (TR2 &4), vitamin-D and retinoid x receptor (VDR-RXR) and oestrogen-receptor 1 (ESR1), which are predicted to be responsible for the differential gene expression observed in docetaxel-resistance. IHC analysis to quantify nuclear expression of the identified TF SRF correlates with both survival from date of bone metastasis (p = 0.003), survival from androgen independence (p = 0.00002), and overall survival from prostate cancer (p = 0.0044). Functional knockdown of SRF by siRNA demonstrated a reversal of apoptotic resistance to docetaxel treatment in the docetaxel-resistant PC-3 cell line model. CONCLUSIONS: Our results suggest that SRF could aid in treatment stratification of prostate cancer, and may also represent a therapeutic target in the treatment of men afflicted with advanced prostate cancer.

Drinking frequency effects on the performance of cattle: a systematic review.

This study used a systematic literature review methodology to determine whether there is evidence that drinking frequency has effects on cattle performance, what performance responses to drinking frequency are documented and how performance responses vary according to environmental and animal factors. Electronic databases were searched for English language articles with original data on at least one performance attribute (e.g. water intake, feed intake, live weight) of cattle in response to voluntary drinking frequency or controlled access periods to water. Sixteen experiments on dairy cows and 12 experiments on beef cattle were retrieved from the literature. For beef cattle, all experiments reported reduced water and feed intake with access to water once every second and/or third day compared with once-daily access. Median reductions of 15% and 25% in water intake and 16% and 9% in feed intake were found across experiments respectively. Live weight responses of beef cattle to access to water were limited and yielded positive, negative and no effects. For dairy cows, most experiments reported reduced water intake, milk yield and milk fat content with access to water twice or once daily compared with controls (ad libitum or ad libitum except at the dairy). Median reductions of 13% and 12% in water intake, 2% and 1% in milk yield and 1% and 2% in milk fat content were found across experiments respectively. Water availability effects on feed intake and live weight were very limited for dairy cows and yielded positive, neutral and negative effects. Season, climate, experiment conditions, animal class and animal genotype were identified to potentially influence intake responses of cattle. The review highlights a number of important gaps in the literature where future work is required to better understand the optimum drinking frequency of cattle and implications of water availability on health, welfare and performance.

Bcl-3 deficiency protects against dextran-sodium sulphate-induced colitis in the mouse.

Bcl-3 is a member of the IκB family of proteins and is an essential negative regulator of Toll-like receptor-induced responses. Recently, a single nucleotide polymorphism associated with reduced Bcl-3 gene expression has been identified as a potential risk factor for Crohn's disease. Here we report that in contrast to the predictions of single nucleotide polymorphism (SNP) analysis, patients with Crohn's disease and ulcerative colitis demonstrate elevated Bcl-3 mRNA expression relative to healthy individuals. To explore further the potential role of Bcl-3 in inflammatory bowel disease (IBD), we used the dextran-sodium sulphate (DSS)-induced model of colitis in Bcl-3(-/-) mice. We found that Bcl-3(-/-) mice were less sensitive to DSS-induced colitis compared to wild-type controls and demonstrated no significant weight loss following treatment. Histological analysis revealed similar levels of oedema and leucocyte infiltration between DSS-treated wild-type and Bcl-3(-/-) mice, but showed that Bcl-3(-/-) mice retained colonic tissue architecture which was absent in wild-type mice following DSS treatment. Analysis of the expression of the proinflammatory cytokines interleukin (IL)-1ß, tumour necrosis factor (TNF)-α and IL-6 revealed no significant differences between DSS-treated Bcl-3(-/-) and wild-type mice. Analysis of intestinal epithelial cell proliferation revealed enhanced proliferation in Bcl-3(-/-) mice, which correlated with preserved tissue architecture. Our results reveal that Bcl-3 has an important role in regulating intestinal epithelial cell proliferation and sensitivity to DSS-induced colitis which is distinct from its role as a negative regulator of inflammation.

Methane emissions from grazing cattle using point-source dispersion.

The ability to accurately measure greenhouse gas (GHG) emissions is essential to gauge our ability to reduce these emissions. Enteric methane from ruminants is an important but often difficult source to quantify since it depends on the amount and type of feed intake. Unfortunately, many of the available measurement techniques for estimating enteric methane emissions can impose a change in feed intake. Our study evaluates a nonintrusive technique that uses a novel approach (point-source dispersion with multiple open-path concentrations) to calculate enteric methane emissions from grazing cattle, reported as the major source of GHG in many countries, particularly Australia. A scanner with a mounted open-path laser was used to measure methane concentration across five paths above a paddock containing 18 grazing cattle over 16 d. These data were used along with wind statistics in a dispersion model (WindTrax) to estimate an average herd methane emission rate over 10-mm intervals. Enteric methane emissions from the herd grazing a combination of Rhodes grass (Chlotis gayana Kunth) and Leucaena [Leucaena leucocephala (Lam.)] averaged (+/- SD) 141 (+/- 147) g animal(-1) d(-1). In a release-recovery experiment, the technique accounted for 77% of the released methane at a single point. Our study shows the technique generates more reliable methane emissions during daytime (unstable stratification).

An assessment of Walk-over-Weighing to estimate short-term individual forage intake in sheep.

The main limitation for determining feed efficiency of freely grazing ruminants is measurement of daily individual feed intake. This paper describes an investigation that assessed a method for estimating intake of forage based on changes in BW of ewes. A total of 24 dry and non-pregnant Romane ewes (12 hoggets, HOG; mean±SD 51.8±2.8 kg BW; body condition score (BCS) 2.6±0.2; and 12 adults, ADU; 60.4±8.5 kg BW; BCS 2.7±0.8) were selected for the study and moved from their rangeland system to a confined pen with controlled conditions and equipped with individual automatic feeders. The experiment lasted for 28 days (21 days adaptation and 7 days feed intake measurement). Ewes were fed hay and trained to use the electronic feeders (one feeding station per ewe) in which actual daily intake (H intake24) was measured. The pens were designed to maximize movement of trained ewes through an automated Walk-over-Weighing device, by using water and mineral salts as attractants. Total individual intake of hay measured in the automatic feeder at each meal (H intake) was compared with indirect estimates of feed intake determined using differences in the BW of the ewes (∆BW) before and 1 h following morning and afternoon feeding at fixed times. The BW, BCS, H intake, H intake24, as well as plasma non-esterified fatty acids (NEFA), glucose and insulin profiles were determined. The BW was higher in ADU v. HOG but BCS was not affected by parity. The H intake24 was affected by day of experiment as a consequence of reduced availability and intake of water on one day. Plasma glucose, NEFA and insulin were not affected by parity or day of experiment. The H Intake was and ∆BW tended to be higher in the morning in HOG, whereas H intake was and ∆BW tended to be higher in ADU at the afternoon meal. Irrespective of parity or feeding time, there was very strong correlation (r 2=0.93) between H intake and ∆BW. This relationship confirms that our indirect method of estimating individual forage intake was reliable within the strictly controlled conditions of the present experiment. The method appears suitable for use in short-term intensive group feeding situations, and has potential to be further developed for longer-term forage intake studies, with a view to developing a method for freely grazing ruminants.

Adrenal venography and ultrasound in the investigation of the adrenal gland: an analysis of 58 cases.

Adrenal venography has been carried out in 58 patients with the left adrenal vein being successfully catheterized in 91 per cent and the right in 77 per cent. Of the 30 patients with primary hyperaldosteronism, 11 adenomas (12-35 mm diameter) have been demonstrated at venography and two of 15 mm suspected, all of which were confirmed surgically. Aldosterone levels in the adrenal vein plasma were raised on the affected side. In the group of proved micronodular hyperplasia, two patients had surgically confirmed macronodules and venography demonstrated one of 12 mm diameter. Two adenomas of 11 mm and one macronodule of 15 mm have been demonstrated at venography in the remainder who have not had an operation. Ultrasound was carried out in 12 patients with primary hyperaldosteronism, ten of which had tumours at venography. Two adenomas measuring 30 and 31 mm were outlined by ultrasound and confirmed surgically. Seven adenomas, including one macronodule (10-25 mm in diameter) were not defined. Three intra-adrenal phaeochromocytomas (45-90 mm) and one extra-adrenal (80-85 mm) were demonstrated at arteriography, identified by ultrasound and confirmed surgically. Of the ten patients with Cushing's syndrome three had enlarged glands at venography, this was confirmed surgically. Cumulative experience from this analysis and published reports indicate that venography will demonstrate tumours of 10 mm or more in diameter and outline enlarged glands; aldosterone assays will lateralize tumours as small as 3 mm; ultrasound will outline tumours of 30 mm and selective adrenal arteriography will demonstrate tumours of 10 mm. One patient developed acute adrenal cortical insufficiency with intra-adrenal extravasation on one side and thrombosis of the central vein on the opposite side. A second case developed temporary adreno-cortical insufficiency. Published reports indicate that the risk of complication is about 1 per cent. The report includes an anatomical study of the efferent adrenal veins in 50 patients paying particular attention to the diameter, number of accessory hepatic veins, and the angle of entry and position of the right adrenal vein.

Evaluation of the processing functions of a digital subtraction angiography image processor.

A large number of imaging systems for digital subtraction angiography (DSA) are now commercially available. Numerous evaluations of the performance of these systems have been reported in the literature. However systematic evaluations of the processing functions of DSA image processors have not been widely reported. Such an evaluation for one commercial system is presented in this paper. Functions evaluated include linear transformation, logarithmic transformation, integration, subtraction and temporal filtration. The observations indicate that image processing results are frequently achieved by indirect routes which compromise the fidelity of the image data.

Percussion of the chest re-visited: a comparison of the diagnostic value of ausculatory and conventional chest percussion.

Percussion of the chest is thought to be insensitive in detecting small or deeply situated chest lesions. A newer technique, ausculatory percussion, has been reported as having a far higher sensitivity. In a controlled blind study the diagnostic value of conventional chest percussion compared with ausculatory percussion was defined using the chest x-ray as a gold standard. The prevalence of disease in 100 lung fields was 26%. The majority of lesions were not detected by either technique resulting in very low sensitivities of 15.4% for conventional percussion and 19.2% for ausculatory percussion. A positive result with ausculatory percussion was twice as likely to be false as true (positive predictive value 31.2%). The technique did not add to the diagnostic value of conventional percussion. Both techniques have major limitations. Patients with suspected lung disease still require chest x-ray examination if percussion is normal.

Endoscopic treatment of urinary tract calculi.

Removal of urinary tract calculi by endourological techniques is now widely practised and has reduced the indications for open surgical removal to a minority of cases. A review of 270 consecutive patients who presented to the Meath Hospital Urology department with urinary tract calculi and were treated by percutaneous or transurethral endoscopy is herein reported. The rates of successful clearance of renal (95%) and ureteric (78%) calculi compare well with previous reports. The majority of complications were minor and a low overall complication rate was experienced at ureterorenoscopy. In our experience, endourological treatment of urinary tract calculi is successful, efficient and safe and is the recommended approach to urinary tract calculi that require surgical intervention.

The role of rigid and hinged polypropylene ankle-foot-orthoses in the management of cerebral palsy: a case study.

Ankle-foot orthoses are commonly used in the treatment of spastic cerebral palsy to hold the foot in a position conductive to a more functional gait. This study, utilizing quantitative biomechanical techniques, evaluates the effects of a rigid ankle-foot orthosis and a hinged ankle-foot orthosis on spastic cerebral palsy gait. The subject was a 4.5 year old female diagnosed as spastic diplegic cerebral palsied shortly after birth. Testing involved collection of kinematic coordinate data employing a WATSMART video system and ground reaction force data using a Kistler force plate. Jensen's (1978) photogrammetric method was used to estimate body segment inertial parameters. The hinged ankle-foot orthosis was found to be more effective than the rigid ankle-foot orthosis. The subject exhibited a more natural ankle motion during the stance phase of gait, greater symmetry of segmental lower extremity motion, and decreased knee moments during stance while wearing a hinged ankle-foot orthosis.

The role of the contralateral limb in below-knee amputee gait.

Very little quantitative biomechanical research has been carried out evaluating issues relevant to prosthetic management. The literature available suggests that amputees may demonstrate an asymmetrical gait pattern. Furthermore, studies suggest that the forces occurring during amputee gait may be unequally distributed between the contralateral and prosthetic lower limbs. This study investigates the role of the contralateral limb in amputee gait by determining lower limb joint reaction forces and symmetry of motion in an amputee and non-amputee population. Seven adult below-knee amputees and four non-amputees participated in the study. Testing involved collection of kinematic coordinate data employing a WATSMART video system and ground reaction force data using a Kistler force plate. The degree of lower limb symmetry was determined using bilateral angle-angle diagrams and a chain encoding technique. Ankle, knee and hip joint reaction forces were estimated in order to evaluate the forces acting across the joints of the amputee's contralateral limb. The amputees demonstrated a lesser degree of lower limb symmetry than the non-amputees. This asymmetrical movement was attributed to the inherent variability of the actions of the prosthetic lower limb. The forces acting across the joints of the contralateral limb were not significantly higher than that of the non-amputee. This suggests that, providing the adult amputee has a good prosthetic fit, there will not be increased forces across the joints of the contralateral limb and consequently no predisposition for the long-term wearer to develop premature degenerative arthritis.

Using UHF proximity loggers to quantify male-female interactions: a scoping study of estrous activity in cattle.

Reproductive efficiency is an important determinant of profitable cattle breeding systems and the success of assisted reproductive techniques (ART) in wildlife conservation programs. Methods of estrous detection used in intensive beef and dairy cattle systems lack accuracy and remain the single biggest issue for improvement of reproductive rates and such methods are not practical for either large-scale extensive beef cattle enterprises or free-living mammalian species. Recent developments in UHF (ultra high frequency) proximity logger telemetry devices have been used to provide a continuous pair-wise measure of associations between individual animals for both livestock and wildlife. The objective of this study was to explore the potential of using UHF telemetry to identify the reproductive cycle phenotype in terms of intensity and duration of estrus. The study was conducted using Belmont Red (interbred Africander Brahman Hereford-Shorthorn) cattle grazing irrigated pasture on Belmont Research Station, northeastern Australia. The cow-bull associations from three groups of cows each with one bull were recorded over a 7-week breeding season and the stage of estrus was identified using ultrasonography. Telemetry data from bull and cows, collected over 4 8-day logger deployments, were log transformed and analyzed by ANOVA. Both the number and duration of bull-cow affiliations were significantly (P<0.001) greater in estrous cows compared to anestrus cows. These results support the development of the UHF technology as a hands-off and noninvasive means of gathering socio-sexual information on both wildlife and livestock for reproductive management.

Natural killer cells protect mice from DSS-induced colitis by regulating neutrophil function via the NKG2A receptor.

Natural killer (NK) cells are traditionally considered in the context of tumor surveillance and infection defense but their role in chronic inflammatory disorders such as inflammatory bowel disease is less clear. Here, we investigated the role of NK cells in dextran sodium sulfate (DSS)-induced colitis in mice. Depletion of NK cells impairs the survival of mice with colitis and is linked with dramatic increases in colonic damage, leukocyte infiltration, and pro-inflammatory profiles. Mice depleted of NK cells had increased numbers of neutrophils in colons and mesenteric lymph nodes, compared with control mice, in addition to acquiring a hyper-activation status. In vitro and in vivo studies demonstrate that NK cells downregulate pro-inflammatory functions of activated neutrophils, including reactive oxygen species and cytokine production, by direct cell-to-cell contact involving the NK cell-inhibitory receptor NKG2A. Our results indicate an immunoregulatory mechanism of action of NK cells attenuating DSS-induced colitis neutrophil-mediated inflammation and tissue injury via NKG2A-dependent mechanisms.

Gene silencing of TNF-alpha in a murine model of acute colitis using a modified cyclodextrin delivery system.

Inflammatory bowel disease (IBD) is a chronic relapsing inflammation of the gastrointestinal tract. The cytokine TNF-alpha (TNF-α) plays a pivotal role in mediating this inflammatory response. RNA interference (RNAi) holds great promise for the specific and selective silencing of aberrantly expressed genes, such as TNF-α in IBD. The aim of this study was to investigate the efficacy of an amphiphilic cationic cyclodextrin (CD) vector for effective TNF-α siRNA delivery to macrophage cells and to mice with induced acute-colitis. The stability of CD.siRNA was examined by gel electrophoresis in biorelevant media reflecting colonic fluids. RAW264.7 cells were transfected with CD.TNF-α siRNA, stimulated with lipopolysaccharide (LPS) and TNF-α and IL-6 responses were measured by PCR and ELISA. Female C57BL/6 mice were exposed to dextran sodium sulphate (DSS) and treated by intrarectal administration with either CD.siRNA TNF-α or a control solution. In vitro, siRNA in CD nanocomplexes remained intact and stable in both fed and fasted simulated colonic fluids. RAW264.7 cells transfected with CD.TNF-α siRNA and stimulated with LPS displayed a significant reduction in both gene and protein levels of TNF-α and IL-6. CD.TNF-α siRNA-treated mice revealed a mild amelioration in clinical signs of colitis, but significant reductions in total colon weight and colonic mRNA expression of TNF-α and IL-6 compared to DSS-control mice were detected. This data indicates the clinical potential of a local CD-based TNF-α siRNA delivery system for the treatment of IBD.

Carbonic anhydrase IX expression in prostate cancer.

Tumour hypoxia is associated with over 70% of solid tumours including prostate and colorectal cancer. Hypoxia promotes tumour progression and resistance to treatment. Carbonic anhydrase IX (CA IX) is an endogenous marker of hypoxia. It is expressed in lung and renal cell carcinomas and is associated with a poor prognosis. CA IX has an important role in maintaining pH levels in the highly metabolically active cancer cell. The expression of CA IX in prostate cancer has not previously been investigated. Immunohistochemistry was used to examine CA IX expression in 59 patients, using tissue microarrays (TMAs) and full sections of BPH, surrounding stroma and prostate adenocarcinoma. Cores reviewed included 189 BPH, 130 Gleason grade 3, 93 Gleason grade 4, 40 Gleason grade 5. CA IX expression in colorectal cancer and HIF 1alpha in prostate cancer acted as positive controls. There was only occasional cell staining for CA IX expression. Although prostate cancer is a hypoxic tumour it does not express CA IX. This implies it relies on alternative pathways for maintaining pH balance in cancer. These studies would indicate that CA IX is not a suitable marker of hypoxia in prostate cancer.

Radio-isotope bone scanning in suspected osteomyelitis in children.

The usefulness of radio-isotope bone scanning in suspected osteomyelitis has been widely acclaimed. Fourteen children had rectilinear bone scans performed three hours after injection of Tc99m methylene diphosphonate. A diagnostic accuracy of 56% was achieved, which is lower than in other series. The reasons for this are discussed and the value of bone scanning in the evaluation of osteomyelitis is questioned.

A solid sorbent personal sampling method for the determination of acrolein in air.

A personal air sampling method, using activated carbon with subsequent analysis by gas chromatography, has been developed for acrolein in the 0.05 to 5 ppm (parts per million) range for a five liter air sample. The sensitivity is 0.25 microgram (microgram) of acrolein. This first known viable method for acrolein using a solid adsorbent was made possible by the discovery that acrolein can be adsorbed on, and recovered from, hydroquinone-treated carbon with efficiencies as high as 90 percent. Samples must be analyzed the same day or frozen and analyzed within five days. Airborne contaminants possibly present in the acrolein production area do not interfere with the analysis.

Bronchial adenoma: review of 18-year experience at the Brompton Hospital.

Continued uncertainty about the prognosis for patients with bronchial adenomata led to a review of the experience of this condition in the Brompton Hospital. Of 72 patients seen between January 1955 and December 1972, 39 were women and 33 men, mean age 45 years, range 9-73 years. The commonest presenting symptoms were haemoptysis, cough, sputum, and repeated chest infections. Positive bronchoscopic biopsy occurred in 35 of 43 cases; five of these were originally reported as carcinomata, of oat-cell type in four. Plain chest film abnormality occurred in 69 patients. Seventy-three operative procedures comprised two endoscopic removals, two wedge resections, six bronchotomies, five pneumonectomies, and 58 lobectomies (seven with sleeve resection). Recurrence in three of six bronchotomies--two with adenoid cystic carcinomata (cylindromata)--necessitated further surgery. Lobectomy and lymph node dissection is usually the operation of choice. Histology confirmed 67 carcinoids (eight with atypical histology or lymph node metastases), two adenoid cystic carcinomata, one muco-epidermoid, and two mucous gland adenomata. Prolonged follow-up is especially indicated in patients with adenoid cyst carcinoma and in those with atypical or metastatic carcinoid histology. Although such pathology is not incompatible with long survival, of 10 patients in these categories, all five late deaths were probably related to the tumour. However, of 57 patients considered to have had typical carcinoid histology and adequate removal of the tumour, there has to date been no tumour-related death, but one patient developed radiosensitive atypical carcinoid tracheal tumours nine years later. The actuarially assessed survival of 71 patients undergoing surgery for bronchial adenomata was 75% at 15 years. Specific tumour types should replace the term bronchial adenoma.