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PubMed1996

IS-RT-PCR assay detection of MT-MMP in a human breast cancer cell line.

The in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR) is a method that allows the direct localisation of gene expression. The method utilises the dual buffer mediated activity of the enzyme rTth DNA polymerase enabling both reverse transcription and DNA amplification. Labelled nucleoside triphosphates allow the site of expression to be labelled, rather than the PCR primers themselves, giving a more accurate localisation of transcript expression and decreased background than standard in situ hybridisation (ISH) assays. The MDA-MB-231 human breast carcinoma (HBC) cell line was assayed via the IS-RT-PCR technique, using primers encoding MT-MMP (membrane-type matrix metalloproteinase) and human beta-actin. Our results clearly indicate baseline expression of MT-MMP in the relatively invasive MDA-MB-231 cell line at a signal intensity similar to the housekeeping gene beta-actin, and results following induction with Concanavalin A (Con A) are consistent with our previous results obtained via Northern blotting.

Breast Neoplasms↗

PubMed2002

Recent developments in contraceptive implants at the Population Council.

Development of contraceptive implant methods, when based on established or on new synthetic chemical entities, is a decadal or multi-decadal process. The process often requires the cooperation of numerous investigators for laboratory work, for early Phase II trials, for dose-finding trials, and for Phase III clinical trials. The Phase III work also requires cooperation with a commercial manufacturer and potential distributor of the product. The Population Council has recently completed developmental work on two levonorgestrel-releasing implants, with filings to regulatory agencies that support extended use of Jadelle implants for 5 years and Norplant implants for 7 years. When the developmental process includes establishing the clinical properties of a molecule not yet approved by regulatory agencies, the minimum development time appears to be two decades. The status and rationale of studies of a new Nestorone-releasing, single implant developed by the Population Council for a period of use of 2 years are presented.

Academies and Institutes↗

PubMed1992

Methods for testing impairment of driving due to drugs.

The Transport and Road Research Laboratory has been concerned for a long time with possible causes of driving difficulties and has developed methods for investigating driving performance. The question addressed here was how applicable these methods are in assessing driving problems arising from the use of drugs which can impair performance, particularly widely-available centrally-acting drugs. We assessed four types of driving-related tests by comparing their sensitivities with two laboratory tests, developed elsewhere, which measure more basic effects of drugs on performance, using drugs known to impair skills. Performances under the influences of ethanol, the benzodiazepine lorazepam, and the antihistamine triprolidine, each given both as a single high dose and a single low dose, were compared with performances after placebo. We used double-blind crossover design, in which subject variability was minimized by studying only women of a limited age range (45-55 y). The driving-related tests detected the effects of the substances used, although they were generally less sensitive than the laboratory tests. The individual sensitivities of the driving test could be improved to match those used for more general assessments.

Automobile Driving↗

PubMed1996

Investigation of anal function in patients with systemic sclerosis.

OBJECTIVE: To investigate anorectal function in women patients with systemic sclerosis (SSc), with and without lower gastrointestinal symptoms. METHODS: Anorectal manometry was performed in 16 patients with SSc: six with no or minimal bowel symptoms, seven with constipation, and three with diarrhoea and faecal incontinence. Eleven healthy women acted as control subjects. Pressure data were recorded via an eight lumen polyvinylchloride water perfused catheter. Station and rapid pull through techniques were used. RESULTS: In the patients with SSc, mean resting pressure, maximal voluntary squeeze effort, and squeeze vector volume were lower, and squeeze asymmetry was greater, compared with the healthy controls. Differences were significant in the subgroup with constipation. CONCLUSION: Radial asymmetry and vector volume parameters provide detailed analysis of segmental anal canal function. Our findings suggest significant segmental deficits in those patients with SSc who have lower gastrointestinal symptoms. The trend towards smaller pressures and squeeze vector volumes in the asymptomatic SSc group suggests subclinical dysfunction in these patients.

Adult↗

PubMed2002

Endoscopic ultrasound-guided gallbladder bile aspiration in idiopathic pancreatitis carries a significant risk of bile peritonitis.

BACKGROUND: Direct microscopic examination of bile for the presence of microlithiasis is often performed during the evaluation of patients with idiopathic pancreatitis. Bile sampled from the duodenum and/or the common bile duct may not represent gallbladder bile, and thus may be inadequate for the diagnosis of microlithiasis. AIM: We sought to determine the safety and efficacy of endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) of gallbladder bile in patients with idiopathic pancreatitis. METHODS: Patients with idiopathic pancreatitis underwent EUS with a linear echoendoscope. After excluding potential causes of pancreatitis such as common bile duct stones or pancreatic lesions, the gallbladder was identified. The gallbladder lumen was entered using a 22-gauge FNA needle via the duodenal wall. Bile was aspirated and analyzed for the presence of cholesterol monohydrate crystal, calcium bilirubinate granules, calcium carbonate microspheroliths and mucin gel strands. RESULTS: Three patients underwent EUS-guided FNA of gallbladder bile. Two of these patients developed bile peritonitis within one hour of the procedure prompting us to discontinue the study. One patient's gallbladder bile contained microlithiasis. This patient had bile aspirated from the common bile duct via ERCP 3 weeks prior to EUS. However, analysis of that bile sample failed to show microlithiasis. CONCLUSION: Unfortunately, transduodenal EUS-guided FNA of gallbladder bile using a 22-gauge needle carries a significant risk of bile peritonitis.

Acute Disease↗

PubMed2006

Body mass index, waist circumference and quality of life in individuals with schizophrenia.

OBJECTIVE: The primary objective was to examine the differential relationship between waist circumference, body mass index, and self-reported quality of life in patients with schizophrenia. METHOD: Individuals with DSM-IV schizophrenia (n=90) were interviewed to obtain sociodemographic data, complete a Quality of Life questionnaire (the MOS SF-12) and have measurements taken of height, weight (kg), and waist circumference (cm). Multiple regression analysis was used to assess the associations between adiposity measures (BMI, WC) and quality of life outcomes (PCS, MCS). RESULTS: Mental component score (MCS) was not significantly related to either of the weight related measures. After adjustment for gender and age, both BMI alone and WC alone were significant predictors of PCS. When both BMI and WC were included in the same regression model, only WC remained a significant predictor of PCS. CONCLUSIONS: Quality of life in schizophrenic patients is related to measures of body weight. The relationship is strongest using waist circumference as the primary measure. This provides further support for routinely incorporating this measure within research and clinical assessments.

Adult↗

PubMed2003

Safety and efficacy of the supraclavicular approach to thoracic outlet decompression.

BACKGROUND: Thoracic outlet syndrome (TOS) is a clinical diagnosis encountered by both thoracic and vascular surgeons. The goal of surgical therapy involves relieving compression of the neurovascular structures at the superior thoracic aperture. The traditional approach to thoracic outlet decompression has been transaxillary; however more centers are moving toward a more tailored approach through a supraclavicular incision. METHODS: The medical records of 67 patients who underwent surgical decompression between 1993 and 2001 for TOS were retrospectively reviewed. Patient demographics and early outcome were assessed through clinic follow-up. RESULTS: Seventy-two thoracic outlet decompressions were performed on 67 patients with the diagnosis of TOS. Five patients underwent bilateral thoracic outlet decompression. All operations in this time period were safely accomplished through a supraclavicular approach. The syndromes associated with thoracic outlet compression were neurogenic (n = 59), venous (n = 10), and arterial (n = 3). Forty-six of 72 (63.9%) operations resulted in complete resolution of symptoms, 17 cases (23.6%) had partial resolution, and 9 patients (12.5%) had no resolution. There were no deaths and morbidity was minimal with 6 complications (8.3%). CONCLUSIONS: The supraclavicular approach is a safe and effective technique in managing all forms of thoracic outlet compression.

Adult↗

PubMed2003

Pyrrolidine dithiocarbamate reduces lung reperfusion injury.

BACKGROUND: The inflammatory cascade has emerged as the primary mediator of reperfusion injury. Nuclear factor kappaB (NF-kappaB) is a rapid response transcription factor that activates genes responsible for the mediators of inflammation. Heat shock protein 70 (HSP70) has been shown to protect against lung injury. We hypothesized that the antioxidant pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB and upregulator of HSP70, would decrease lung injury after ischemia and reperfusion. METHODS: Using our isolated, ventilated, blood-perfused rabbit lung model, all groups underwent lung harvest followed by 10-h storage (4 degrees C) and blood reperfusion for 30 min. Group I lungs (n = 5) served as controls. In group II lungs (n = 5), both lung and blood donors received PDTC (100 mg/kg) intravenously 30 min before harvest. NF-kappaB activity was evaluated with electrophoretic mobility shift assay, and Western blot was performed for HSP70. RESULTS: Group II demonstrated superior pulmonary function. Although HSP70 expression was somewhat elevated in group II lungs, NF-kappaB binding activity was not different between the groups. CONCLUSIONS: PDTC improves pulmonary function after ischemia and reperfusion in an isolated rabbit lung model. The improved function correlates with elevated HSP70 expression during initial reperfusion, independent of NF-kappaB activity.

Animals↗

PubMed2006

High-frequency probe ultrasonography has limited accuracy for detecting invasive adenocarcinoma in patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma: a case series.

AIM: To evaluate prospectively the accuracy of preoperative high-frequency (20 MHz) probe ultrasonography (HFPUS) for detecting invasive cancer in patients referred for esophagectomy because of an endoscopic biopsy diagnosis of high-grade dysplasia (HGD) or intramucosal carcinoma (ICA) in Barrett's esophagus (BE). PATIENTS AND METHODS: Nine consecutive male patients (median age of 69 yr) who were referred for esophagectomy for HGD or ICA in BE agreed to participate. We performed conventional upper gastrointestinal endoscopy followed by HFPUS using a through-the-scope ultrasound probe (20 MHz), and we compared our preoperative findings with the pathologist's findings in the resected esophageal specimens. RESULTS: There was complete agreement between the postoperative pathological findings and the preoperative HFPUS findings in only 4 of the 9 patients. HFPUS resulted in two false-negative diagnoses of esophageal cancer (both had T1 lesions in the resected specimens), one false-positive diagnosis of esophageal cancer, and two errors in tumor staging (1 understaged, 1 overstaged). CONCLUSIONS: HFPUS has limited accuracy for identifying invasive cancer in patients found to have HGD or IMC in BE. Pending further refinements in technology, clinical management decisions in such patients should not be based solely on the results of HFPUS.

Adenocarcinoma↗

PubMedDate not supplied

Real-time computed tomography dosimetry during ultrasound-guided brachytherapy for prostate cancer.

PURPOSE: Ultrasound-guided implantation of permanent radioactive seeds is a treatment option for localized prostate cancer. Several techniques have been described for the optimal placement of the seeds in the prostate during this procedure. Postimplantation dosimetric calculations are performed after the implant. Areas of underdosing can only be corrected with either an external beam boost or by performing a second implant. We demonstrate the feasibility of performing computed tomography (CT)-based postplanning during the ultrasound-guided implant and subsequently correcting for underdosed areas. METHODS AND MATERIALS: Ultrasound-guided brachytherapy is performed on a modified CT table with general anesthesia. The postplanning CT scan is performed after the implant, while the patient is still under anesthesia. Additional seeds are implanted into "cold spots," and the resultant dosimetry confirmed with CT. RESULTS: Intraoperative postplanning was successfully performed. Dose-volume histograms demonstrated adequate dose coverage during the initial implant, but on detailed analysis, for some patients, areas of underdosing were observed either at the apex or the peripheral zone. Additional seeds were implanted to bring these areas to prescription dose. CONCLUSION: Intraoperative postplanning is feasible during ultrasound-guided brachytherapy for prostate cancer. Although the postimplant dose-volume histograms for all patients, before the implantation of additional seeds, were adequate according to the American Brachytherapy Society criteria, specific critical areas can be underdosed. Additional seeds can then be implanted to optimize the dosimetry and reduce the risk of underdosing areas of cancer.

Equipment Design↗

PubMed1986

Magnetic resonance imaging--first human images in Australia.

The use of magnetic resonance imaging, in the demonstration of internal human anatomy and in the diagnosis of disease, has the major advantages that the technique is noninvasive, does not require the use of ionizing radiation and that it can demonstrate neurological and cardiovascular lesions that cannot be diagnosed easily by other imaging methods. Magnetic resonance imaging is derived from the principle that certain atomic nuclei in a strong magnetic field will absorb pulses of radiofrequency energy; when the pulse is finished the nuclei will emit radiowaves at the same frequency. These radiowaves are received by specially designed aerials or coils and the information is collected by a computer which reconstructs an image of internal anatomy in a similar way to that of x-ray computed tomography (CT). By changing the strength of the magnetic fields and the frequency of the radiowave pulses, it is possible to examine different sections within the body. The first magnetic resonance images of humans were obtained in Australia in October 1985 on the research instrument of the Queensland Medical Magnetic Resonance Research Centre, which is based at the Mater Hospital in Brisbane, and is part of the University of Queensland's Department of Radiology.

Abdomen↗

PubMed1996

Steady-state polarization from cylindrically symmetric fluorophores undergoing rapid restricted motion.

Steady-state fluorescence polarization measurements provide a relatively simple method for investigating the orientation of molecular components in ordered biological systems. However, the observed fluorescence polarization ratios also depend on any mobility of the fluorophores on the time scale of the fluorescence lifetime. Such mobility commonly arises from incomplete immobilization of a fluorescent probe on the macromolecule of interest. A theoretical formalism is presented for the interpretation of steady-state fluorescence polarization ratios in the presence of such rapid fluorophore motion. It is assumed that the fluorophores move freely within a cone between absorption and emission of a photon. Only one new parameter is introduced to describe fluorophore motion, the semi-angle of the cone, and this can be measured in separate experiments on an isotropic system. The fluorophore orientations are assumed to have cylindrical symmetry, but the symmetry axis need not be in the same plane as the center axes of the absorption and emission cones. This geometry applies to muscle and other biological fibers.

Animals↗

PubMed2004

A novel method for screening viral interferon-resistance genes.

Many viruses have evolved mechanisms to antagonize the interferon (IFN) system, targeting all the major components involved in receptor binding and signaling. Although a number of these vital proteins are homologous to cellular proteins involved in IFN downregulation (e.g., viral IFN regulatory factors [vIRFs]), many share little resemblance to known proteins. To determine the IFN-blocking properties of these proteins, functional assays are required. Here, we present a new and rapid functional screening method, based on the 2fTGH cell line, which is able to determine viral gene products that inhibit the IFN-alpha/Jak-Stat signaling pathway. Expression cloning of viral IFN-blocking genes into 2fTGH and consequent selection with IFN-alpha and 6-thioguanine result in the outgrowth of cells that are no longer responsive to IFN-alpha. We also demonstrate that selection occurs if members of the Jak-Stat signaling pathway are lost. To show the utility of our system, we have used a known suppressor of IFN signaling, the human papillomavirus (HPV) E7 gene. Expression of E7 causes the loss of ability of 2fTGH cells to respond to IFN-alpha treatment because of a functional disruption of the signaling pathway. This approach offers a new strategy for identifying novel viral genes or new functions of already described viral genes that have a role in IFN-alpha signaling inhibition.

Cell Line↗

PubMed2004

Effects of protein calorie malnutrition on the pharmacokinetics of ketamine in rats.

The effect of protein calorie malnutrition (PCM) on the pharmacokinetics of ketamine (KET) enantiomers has been investigated. Six control and six PCM rats were administered 85 mg/kg racemic KET by intramuscular injection, and plasma concentrations of (S)- and (R)-KET, norketamine (NKET), and 5,6-dehydronorketamine (DNK) were measured using enantioselective gas chromatography. Pharmacokinetic profiles were analyzed using standard noncompartmental and compartmental modeling methods. The volumes of distribution were similar between control and PCM rats for (S)- and (R)-KET. However, total clearance of both KET enantiomers was decreased, resulting in an increase in systemic exposure (p < 0.05). The KET absorption rate was also increased in PCM rats. A decrease in the clearance of both NKET enantiomers led to a significant increase in exposure in PCM rats (p < 0.005), and modeling results could not exclude the possibility that PCM induced an increase in the fraction of KET following the NKET pathway, which may further contribute to this increase in exposure. An increase in exposure to DNK enantiomers was also evident in PCM animals compared with controls [p < 0.005 (DNK1); N.S. (DNK2)], which was in concordance with the decrease in apparent clearance values. These results show that PCM significantly alters the pharmacokinetics of KET and several of its metabolites.

Animals↗

PubMed2005

The effects of hypnotic and nonhypnotic imaginative suggestion on pain.

BACKGROUND: Few studies have compared placebo and suggested pain reduction. PURPOSE: Hypnotic and nonhypnotic imaginative analgesia suggestions were compared against a placebo in reducing experimental pain. The mediator role of response expectancies and the moderator role of hypnotic and nonhypnotic imaginative suggestibility were evaluated. METHODS: Sixty participants previously assessed for hypnotic and nonhypnotic imaginative suggestibility were assigned to one of two experimental conditions or a no-treatment control condition. In the "placebo first" condition, participants received placebo, followed by imaginative and then hypnotic analgesia suggestions. In the "placebo last" condition, participants received imaginative and then hypnotic suggestions, followed by placebo. RESULTS: Imaginative and hypnotic suggestions did not differ significantly and were more effective than no treatment in reducing pain. The placebo was no different from the analgesia suggestions and was more effective than no treatment, but only when administered after the suggestions. Pain reduction was mediated by expectancy but was not significantly related to suggestibility or hypnotizability, the latter operationalized as hypnotic suggestibility with imaginative suggestibility statistically controlled. CONCLUSIONS: In the general population, nonhypnotic imaginative suggestions may be as effective as hypnotic suggestions in reducing pain. Response expectancies would seem to be an important mechanism of placebo and suggested pain reduction.

Humans↗

PubMed2002

Governing the conduct of conduct: are restraints inevitable?

BACKGROUND: The purpose of this article is to provide an interpretation of how restraint use is maintained and legitimized despite negative reports on its efficacy and questions about its ethicality. My research examined the use of restraint on a patient requiring care in an acute teaching hospital in Australia. This article examines one case study that was part of a PhD research project. The literature reveals evidence of the harm that restraints cause, as well as their ineffectiveness as a safety measure. In addition, it indicates that the prevalence of restraint use is high. METHODS: The study is framed by a Foucauldian approach to discourse analysis. This report is an in-depth case study including observations of the patient, interviews with members of the multidisciplinary team and analysis of medical, physiotherapy and nursing notation. FINDINGS: Discourses from the health care team are identified by which restraint use is justified, and legitimized by staff. An important discursive practice is 'constituting the patient's inability to self govern' and their resulting marginalization from the services provided by the team. LIMITATIONS: There are certain philosophical limitations including classical arguments about the position of postmodernism and specifically the ideas of Michel Foucault as opposed to other philosophy's ability to make sense of a phenomenon. The study was carried out in Australia and thus the discursive practices may be dissimilar in other countries. CONCLUSIONS: Through these discursive practices we can understand how staff maintain a monopoly over the truth and perpetuate claims about the inevitability of restraint use.

Aged↗

PubMed1999

Extra-anatomic stents in ureteric obstruction: experience and complications.

OBJECTIVE: To assess the role of extra-anatomic stents (EAS) as a means of urinary diversion in patients with ureteric obstruction secondary to malignancy. PATIENTS AND METHODS: The technique for inserting EAS in patients with ureteric obstruction was described previously; to date, 13 patients (seven women and six men, mean age 45.3 years, range 22-78) have been treated. All patients had ultrasonographic evidence of hydronephrosis and/or significant biochemical evidence of renal impairment. Patients had advanced malignancy and one patient an abdominal aortic aneurysm. RESULTS: Urinary diversion was successful in all patients; two survived for more than 1 year, with stent changes at 6-monthly intervals. In three patients the stents were replaced by percutaneous nephrostomies because of problems with leakage or infection. The remaining patients died with functioning EAS in situ. CONCLUSIONS: In patients with ureteric obstruction secondary to malignancy or medical conditions excluding them from more invasive surgery, EAS provide a further therapeutic option instead of a permanent nephrostomy, which has associated inherent problems. This technique is not without potential problems and careful selection of patients remains vital in this difficult area.

Adult↗

PubMed2007

Galectin-3 expression in the intervertebral disc: a useful marker of the notochord phenotype?

STUDY DESIGN: Galectin-3 expression in rat intervertebral disc at different stages in postnatal life is evaluated. OBJECTIVE: To determine if galectin-3 expression is confined to cells of the nucleus pulposus in the postnatal rat intervertebral disc. SUMMARY OF BACKGROUND DATA: During embryonic development, the anulus fibrosus is derived from the sclerotome, whereas the nucleus pulposus is notochordal. Many authorities opine that in the postnatal disc, notochordal cells play a central role in controlling the development of degenerative disc disease. Surprisingly, unequivocal evidence supporting the existence of notochordal cells in the nucleus pulposus in postnatal life has yet to be demonstrated. Since the expression of galectin-3 is commonly used to identify notochordal cells, we evaluated its expression in tissues of the rat disc and in cultured cells. METHODS: Galectin-3 expression was studied in the nucleus pulposus and anulus fibrosus tissue of rat discs (2 days, 9 weeks, and 10 months old), and cultured cells using different biochemical and molecular biology methods. Rat sternal cartilage and cultured sternal chondrocytes were used as controls. RESULTS: Immunohistochemical studies indicated that galectin-3 was present in the nucleus pulposus and anulus fibrosus. In both discal tissues and cultured cells, studies confirmed that there was a robust expression of galectin-3 messenger ribonucleic acid and protein. Protein expression patterns were similar in neonatal, young, and mature rats. There was also evidence of intracellular and membrane expression of galectin-3 in the cultured disc cells. Finally, significant levels of galectin-3 were evident in rat sternal cartilage and cultured sternal chondrocytes. CONCLUSIONS: Results of the study indicate that galectin-3 is expressed in the neonatal, young, and mature rat disc, and its expression is not restricted to the cells of the nucleus pulposus. Because of its ubiquitous expression, this protein cannot be used as a marker of notochordal cells in the postnatal rat disc.

Aging↗