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PubMed2006

HER2 expression and gene amplification in pT2a Gleason score 6 prostate cancer incidentally detected in cystoprostatectomies: comparison with clinically detected androgen-dependent and androgen-independent cancer.

Previous studies have demonstrated HER2 protein overexpression and/or gene amplification in a subset of patients with clinically significant prostate cancer (PCa), especially in the androgen-independent phase of the disease. There are no studies on incidentally detected PCa. The aim of the study was to analyze HER2 expression and gene amplification in PCa incidentally detected in cystoprostatectomies. High-grade prostatic intraepithelial neoplasia (HGPIN) was also investigated. Comparison was made with clinically detected PCa, both untreated and hormonally treated, and with androgen-independent PCa. Nineteen cystoprostatectomy (CyP) and 44 radical prostatectomy specimens (25 untreated and 19 hormonally treated) with pT2a Gleason score 6 cancer and HGPIN were used in this study. It also included 9 specimens of transurethral resection of the prostate with hormone-independent cancer and 8 cases of normal prostate tissue from CyP specimens without PCa and prostatic intraepithelial neoplasia. HER2 protein and Ki-67 were investigated immunohistochemically. Patients with immunohistochemical scores of 2+ and 3+ were considered to have HER2 overexpression (HercepTest method). Dual-color fluorescence in situ hybridization analysis was performed using the CEP-17/HER dual probe combination. High-grade prostatic intraepithelial neoplasia showed HER2 overexpression in 26% of the CyP cases and in 40% and 83% of the untreated and treated cases, respectively. Prostate cancer showed HER2 overexpression in 16% of cases in the CyP group and in 36% and 47.5% in the untreated and treated groups, respectively. HER2 overexpression was present in 78% of androgen-independent cancers. HER2 gene amplification was seen in a small proportion of nuclei and some of the cases. In HGPIN, it ranged from 1.1% (in 5 cases) in the CyP group to 2.1% (in 10 cases) and 1.9% (in 6 cases) in the untreated and treated groups, respectively. In PCa, the proportion of nuclei with gene amplification was 0.7% (in 3 cases) in the CyP group, 2.6% (in 10 cases) and 2.5% (in 12 cases) in the untreated and treated groups, respectively, and 9% (in 6 cases) in the androgen-independent PCa. Ki-67 expression in HGPIN and PCa in CyP specimens was lower than in the radical prostatectomies and cases of transurethral resection of the prostate. Our findings in the current HER2-related study indicate that incidentally detected cancer has features of less aggressiveness than clinically detected cancer. This may contribute to a better understanding of the results obtained in screening programs where insignificant cancers are detected along with clinically significant cancers.

Adult↗

PubMedDate not supplied

The early detection research network surface-enhanced laser desorption and ionization prostate cancer detection study: A study in biomarker validation in genitourinary oncology.

Prostate-specific antigen (PSA) screening has led to a dramatic increase in prostate cancer detection with a concurrent stage migration. Although the test has revolutionized prostate cancer detection by identifying disease that is potentially curable in the majority of men, only 25% of men receiving test results of PSA > 4 ng/ml will have prostate cancer and many men receiving a normal PSA will have disease, including high-grade disease. There is a need for improved biomarkers for detecting prostate cancer. One such method of cancer detection is surface-enhanced laser desorption and ionization (SELDI). The Early Detection Research Network (EDRN) validation study for SELDI for prostate cancer is described. In a three-stage study, the portability and reproducibility of the technique will be determined; the predictive algorithm will be refined in a multi-institutional case-control population; followed by ultimate validation in the context of a prospective trial with complete disease ascertainment. The unique aspect of the EDRN SELDI validation study is the novel use of two groups of cancer cases: those cases with higher-risk disease (Gleason > or = 7) and those cases with lower-risk disease (Gleason < or = 6). This study will allow the first evaluation of a predictive algorithm that includes prognosis in disease screening. The EDRN SELDI prostate cancer biomarker validation study is a rigorous evaluation of a new detection method for prostate cancer. The methodologies used for this evaluation will prove useful for guiding future biomarker studies in this challenging disease.

Biomarkers, Tumor↗

PubMed1989

Separation and detection of bis(alkylthio)selenides of penicillamine and glutathione by liquid chromatography with electrochemical detection.

Sensitive and selective methodology is described for the simultaneous determination of thiols, disulfides and bis(alkylthio)selenides in mixtures formed by reaction of selenious acid with thiols. The methods are based on separation of the various compounds by reversed-phase ion pairing chromatography followed by detection with an electrochemical detector equipped with dual mercury/gold amalgam electrodes in series in the eluent stream. Thiols are detected directly at the downstream electrode. Bis(alkylthio)selenides and disulfides are detected indirectly by first reducing them at the upstream electrode followed by detection of the reduction products at the downstream electrode. Bis(alkylthio)selenides are reduced at less negative potentials than disulfides, making it possible to selectively detect them in mixtures with disulfides. Detector response to bis(penicillamine)selenides is linear from 1.1.10(-7) M up to at least 1.2.10(-4) M; the detection limit is estimated to be less than 2 pmol bis(penicillamine)selenide.

Chromatography, High Pressure Liquid↗

PubMed1998

Determination of a variety of chemical classes of pesticides in surface and ground waters by off-line solid-phase extraction, gas chromatography with electron-capture and nitrogen-phosphorus detection, and high-performance liquid chromatography with post-column derivatization and fluorescence detection.

Octadecyl (C18)-bonded porous silica was evaluated for the extraction of triazines, organochlorine, carbamates and acidic pesticides from surface and ground water. Gas chromatography with selected detection methods (electron-capture detection, nitrogen-phosphorus detection, mass spectrometry) and liquid chromatography-post-column derivatization fluorescence detection was employed for the determination of 32 pesticides. Recoveries varied from 52 to 102%. The recoveries of triazines obtained using C18 extraction cartridges and conventional liquid-liquid extraction (LLE) are compared. The limit of detection for seventeen organochlorine compounds was better than 0.003 microgram/l and the limit of detection for other 15 analytes was better than 0.06 microgram/l. The proposed analytical methodology was applied to analyze pesticides in surface and ground-water samples of the Lassithi Plateau, Crete, Greece.

Chromatography, Gas↗

PubMed1999

Separation and detection of common mono- and divalent cations by ion chromatography with an ODS column and conductivity/UV detection.

The retention and detection behavior of common mono- and divalent cations (M+, alkali metal (Li+, Na+, K+, Rb+, Cs+) and ammonium ions (NH4+); M2+, alkaline earth metal ions (Mg2+, Ca2+, Sr2+, Ba2+) was examined using an ODS column (150 x 4.6 mm I.D.) and conductivity (CD)/UV detection. The results obtained were as follows: (1) for M+, the mobile phase, 0.1 mM sodium dodecyl sulphate (SDS) + 10 mM HNO3 and indirect CD detection were effective. (2) Addition of Ce(III) in the mobile phase accelerated the elution of both M+ and M2+. The separation of above 10 cations on an ODS column was achieved for the first time without any coelution of cations and disturbance by system peak. Addition of higher SDS resulted in good separation of M+ and M2+ with longer retention times. CD detection was possible for M+ and M2+ and UV detection for M2+. (3) For M2+, the mobile phase, 0.8 mM Ce(III) + 0.1 mM SDS + 1 mM HNO3 and indirect UV detection were effective. The IC methods were applied to real samples.

Cations, Divalent↗

PubMed1994

Prenatally detected posterior urethral valves: is gestational age at detection a predictor of outcome?

Between 1982 and 1992, 67 boys with posterior urethral valves were managed at our hospital, including 32 (48%) in whom the condition was detected prenatally. We examined the relationship between gestational age at detection and outcome at a median followup of 3.9 years (range 4 months to 10 years). Detection at or before 24 weeks of gestation predicted a poor outcome with 9 of 17 patients (53%) dead or in chronic renal failure at followup. Of the cases detected later in pregnancy only 1 had a poor outcome (p = 0.01). All of the cases detected after 24 weeks of gestation had had normal second trimester scans. Growth parameters for boys in the early and late detection groups were not statistically different. However, there was a significant association between renal failure and growth with 67% versus 14% having heights less than the 3rd percentile (p = 0.05). Respiratory distress at birth predicted a poor outcome, while the presence of palpable abdominal abnormalities or vesicoureteral reflux failed to predict outcome.

Female↗

PubMed2000

Detection of HuD transcripts by means of reverse transcriptase and polymerase chain reaction: implications for the detection of minimal residual disease in patients with small cell lung cancer.

Small cell lung cancer (SCLC) expresses neuroectodermal markers including HuD, the best characterized member of the Hu gene family. The aim of this study is to optimize a simple and sensitive reverse transcriptase-polymerase chain reaction assay to detect circulating HuD-expressing cells for the early detection of SCLC recurrences. HuD-specific primers that selectively amplify the three HuD isoforms allowed the detection of one tumor cell/10(6) non-tumor cells. However, HuD transcripts were also detected in the mononuclear fraction of all samples from normal individuals (n=6) and patients with SCLC (n=5). By contrast, HuD protein was not detected in these fractions using Western blotting. More quantitative assays are necessary to examine the value of HuD transcript detection for the identification of tumor recurrences.

Alternative Splicing↗

PubMed2002

Simultaneous detection of Anaplasma and Ehrlichia species in ruminants and detection of Ehrlichia ruminantium in Amblyomma variegatum ticks by reverse line blot hybridization.

The detection of Anaplasma and Ehrlichia species is usually based on species-specific PCR assays, since no assay is yet available which can detect and identify these species simultaneously. To this end, we developed a reverse line blot (RLB) assay for simultaneous detection and identification of Anaplasma and Ehrlichia species in domestic ruminants and ticks. In a PCR the hypervariable V1 region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for members of the genera Anaplasma and Ehrlichia [Int. J. Syst. Evol. Microbiol. 51 (2001) 2145]. Amplified PCR products from blood of domestic ruminants or Amblyomma variegatum tick samples were hybridized onto a membrane to which eight species-specific oligonucleotide probes and one Ehrlichia and Anaplasma catch-all oligonucleotide probe were covalently linked. No DNA was amplified from uninfected blood, nor from other hemoparasites such as Theileria annulata, or Babesia bigemina. The species-specific probes did not cross-react with DNA amplified from other species. E. ruminantium, A. ovis and another Ehrlichia were identified by RLB in blood samples collected from small ruminants in Mozambique. Finally, A. variegatum ticks were tested after feeding on E. ruminantium infected sheep. E. ruminantium could be detected in adult ticks even if feeding of nymphs was carried out 3.5 years post-infection. In conclusion, the developed species-specific oligonucleotide probes used in an RLB assay can simultaneously detect and identify several Ehrlichia and Anaplasma species. However, as no quantitative data for the detection limit are available yet, only positive results are interpretable at this stage.

Anaplasma↗

PubMed1984

Determination of (endogenous) vitamin K1 in human plasma by reversed-phase high-performance liquid chromatography using fluorometric detection after post-column electrochemical reduction. Comparison with ultraviolet, single and dual electrochemical detection.

A new method for the fluorometric detection of K1 and K2 vitamins using post-column electrochemical reduction is described. Dual electrochemical detection in the reduction/reoxidation mode for coulometric/coulometric as well as coulometric/amperometric detection appears to be more sensitive and selective towards the plasma background than simple reductive electrochemical detection, but fluorometric detection after coulometric reduction offers the best results. Combination of normal-phase chromatography and the described method is only possible if supporting electrolyte is added post-column, but leads to higher detection limits. This highly sensitive method is applied to the determination of vitamin K1 in human plasma samples.

Chromatography, High Pressure Liquid↗

PubMed1991

Detection of Escherichia coli rRNA using target amplification and time-resolved fluorescence detection.

The development of technology to increase the sensitivity and speed of detection of bacterial pathogens in samples is important for diagnosis and monitoring of illness. We have developed a sensitive and rapid method for the detection of bacteria, using Escherichia coli as a model, which combines transcription-based target amplification with a bead-based sandwich hybridization assay using rare earth metal chelate labelled probes and time-resolved fluorescence detection. Using these methods as little as 100 copies (0.00016 attomoles) of purified native Escherichia coli rRNA or just one bacterial cell in a spiked sample could be detected. These results demonstrate that amplification of rRNA by transcription-based amplification and detection by time-resolved fluorescence provide a sensitive technology for the direct detection of micro-organisms without the requirement for prior cultivation.

Base Sequence↗

PubMed1995

The sensitive detection of fluorescently labelled PCR products using an automated detection system.

The polymerase chain reaction plays a central role in many detection assays and methods to improve the sensitivity and specificity of these detection systems are constantly being explored. In this study we investigated the use of an automated laser fluorescent system (ALF) in the context of DNA-based diagnostics for pathogenic bacteria. PCR products were generated using species-specific primer sets, one of which was labelled with a 5' fluorescein. PCR products with a fluorescent label were detected on line with an ALF DNA sequencer and the sensitivity of detection was found to be comparable to that for DNA probe hybridization with a radioactive probe. The technology was successfully applied to the detection of Mycobacterium tuberculosis supplemented into sputum samples and to the detection of listeria in paraffin-embedded tissue samples.

Base Sequence↗

PubMed2003

An irrelevant light enhances auditory detection in humans: a psychophysical analysis of multisensory integration in stimulus detection.

Physiological and behavioral studies in animal models have revealed that information from the different senses can be used synergistically to enhance the detection of objects and events. Although a great deal of evidence exists which demonstrates the synergistic use of multisensory cues in human behavior and perception, there is conflicting evidence about whether such interactions can be used to aid in stimulus detection. To examine this issue, we had volunteers indicate the presence or absence of a brief, low-intensity sound that was either presented alone or paired with a simultaneous light in a one-interval signal detection task. In the first experiment, the task-irrelevant light was found to enhance the detectability of the sound, but also to increase the subject's willingness to report the presence of a sound (i.e., response bias). In the second experiment, designed to eliminate this response bias, we found an improvement in stimulus detectability in the absence of any change in bias. These findings demonstrate the presence of a significant multisensory-mediated gain in stimulus detection in human subjects.

Acoustic Stimulation↗

PubMed1999

Detection of herpes simplex virus (HSV) genome using polymerase chain reaction (PCR) in clinical samples comparison of PCR with standard laboratory methods for the detection of HSV.

BACKGROUND: Diagnosis of Herpes simplex virus (HSV) infections is achieved by detecting the antigen and isolating the virus from the specimen, which requires 7-28 days. With the recently introduced molecular biological technique of polymerase chain reaction (PCR), the diagnosis of HSV infections has been made more rapid and specific. OBJECTIVE: We evaluated PCR in comparison with the standard laboratory methods on different types of clinical specimens referred to in our laboratory. STUDY DESIGN: A total of 54 specimens, from 54 patients, were investigated. Antigen detection on direct smears was carried out using fluorescent antibody test (FAT) and virus isolation was performed using conventional tube culture method. PCR was carried out with the DNA extracted from various specimens using primers, which coded for the DNA polymerase gene giving a 179 base pair (bp) product. RESULTS: The primers were specific for HSV-1 and HSV-2, and the sensitivity of the primers was found to be 0.5 and 0.2 fg in the detection of HSV-1 and HSV-2 DNA, respectively. Of the 50 specimens (excluding 4 archival formalin fixed tissue specimens, which were not subjected to virological methods of detection) HSV was detected by virological methods and PCR in nine specimens, and by PCR alone in 15 additional specimens, thus increasing the analytical sensitivity significantly by 30% (McNemar test: P = 0.0001). The positivity of PCR in all nine virologically positive specimens and the 4 archival specimens obtained from proven lesions of HSV infections further confirmed the specificity of the PCR. CONCLUSION: PCR, in our study, was found to be a rapid, specific and highly sensitive method for the detection of HSV in clinical specimens.

Animals↗

PubMed2002

Detection of doxorubicin and metabolites in cell extracts and in single cells by capillary electrophoresis with laser-induced fluorescence detection.

Capillary electrophoresis with laser-induced fluorescence detection was used to separate and detect doxorubicin and at least five metabolites from NS-1 cells that were treated with 25 microM doxorubicin for 8 h. Using 10 mM borate, 10 mM sodium dodecyl sulfate (pH 9.3) as separation buffer, the 488-nm argon-ion laser line for fluorescence excitation, and a 635 +/- 27.5 nm bandpass filter for detection, the limit of detection (S/N=3) for doxorubicin is 61 +/- 13 zmol. This low limit of detection allows for the detection of a larger number of metabolites than previously reported. Two extraction procedures were performed: a bulk liquid-liquid extraction and an in-capillary single-cell lysis. While in the bulk liquid-liquid extraction procedure, recovery for doxorubicin range from 50 to 99%, in single cell analysis the recovery is expected to be complete. Furthermore performing lysis of a single cell inside the separation capillary prevents doxorubicin or metabolite loss or degradation during handling. Based on the bulk method the calculated metabolite abundance is in the sub-amol per cell range while it varies from 0.1 to 1.1 fmol per cell in single cell analysis confirming metabolite loss during handling. Each metabolite was found at a level less than 0.1% of the doxorubicin content in either method, suggesting a slow metabolism in the NS-1 cell system or effective removal of metabolites by the cell.

Antibiotics, Antineoplastic↗

PubMed1989

Comparison of pulsed coulometric detection and potential-sweep pulsed coulometric detection for underivatized amino acids in liquid chromatography.

Pulsed coulometric detection (PCD) and potential-sweep pulsed coulometric detection (PS-PCD) are applied to the direct detection of amino acids in protein hydrolyzates. The detection mechanisms are based upon surface-catalyzed oxidation of the amine functionalities activated by the transient formation of surface oxide on Au electrodes. PCD uses a triple-step potential waveform in which the integration of electrode current at a constant potential is followed by anodic and cathodic polarizations to clean and reactivate the electrode surface. PS-PCD incorporates a cyclic potential sweep into the triple-step waveform which proceeds through the formation and subsequent removal of the surface oxide with simultaneous current integration. Reactions catalyzed by the formation of the noble metal oxide can be monitored in PS-PCD with the automatic rejection of the surface oxide background. A significant decrease is obtained also in the fluctuation and drift in the base line resulting from gradient elution and variations of the electrode surface. PCD and PS-PCD following gradient elution chromatography are demonstrated to allow for the direct detection of 20 amino acids including secondary amino acids. Detection limits for lysine are ca. 220 ppb (11 ng, 75 pmol) by PCD and 60 ppb (3 ng, 19 pmol) by PS-PCD applied at gold electrodes.

Amino Acids↗

PubMed2005

Multidimensional detection of nitroorganic explosives by gas chromatography-pyrolysis-ultraviolet detection.

We describe a new methodology for the trace detection of organic explosives containing nitro functionalities. Conventional gas chromatography separates the components of an explosive mixture. Effluent from the gas chromatograph is pyrolyzed by passage over a heated Nichrome wire. Nitric oxide produced on pyrolysis of a nitroorganic compound is then detected by ultraviolet absorption spectroscopy between 180 and 240 nm, using a deuterium lamp as the light source. Nitric oxide exhibits a sharply banded, characteristic spectrum in this region, enabling detection of nitroorganics. The system is tested using the explosive simulants nitrobenzene and 2,4-dinitrotoluene, and with the nitramine explosive tetryl. Detection limits are 25 ng for nitrobenzene and 50 ng for 2,4-dinitrotoluene. Tetryl is detected with a detection limit of 50 ng. The system is both easy to implement and could be built as a compact, low-power device.

Journal Article↗

PubMed2000

Antimicrobial residue detection in chicken yolk samples following administration to egg-producing chickens and effects of residue detection on competitive exclusion culture (PREEMPT) establishment.

Competitive exclusion (CE) cultures may offer alternatives to antimicrobial agents for disease prophylaxis in poultry. To avoid potential transfer of antibiotic resistance, safe and effective CE cultures must, by necessity, be highly sensitive to antimicrobial residues. The following studies evaluated the effect of maternal administration of selected antibiotics on the establishment of a licensed CE culture, PREEMPT. Selected antibiotics were administered to actively laying hens for a period of 7 days (experiment 1) or 9 days (experiment 2) in drinking water [sulfadimethoxine (0.05%), enrofloxacin (0.005%), and tylosin tartrate (0.05%)] or feed (sulfadimethoxine with ormetoprim, 250 ppm). In experiment 1, fertile eggs were collected daily and subjected to bioassay for detectable antimicrobial residues in yolk. Antimicrobial residues were not detected during the 7 days of treatment or the subsequent 3 days following cessation of treatment in the control, sulfadimethoxine, sulfadimethoxine with ormetoprim, or tylosin treatment groups. However, detectable residues were observed in eggs derived from enrofloxacin-treated hens on days 6 and 7 during antibiotic administration and also on days 2 and 3 post-antibiotic administration. In experiment 2, antimicrobial residues were also only detected in yolks from hens treated with enrofloxacin. Residue detection occurred on days 2-6 of antibiotic administration, on day 9 of antibiotic administration, on days 1-3 post-antibiotic administration, and also on day 7 post-antibiotic administration. A subset of eggs from each experimental group, corresponding to days 2-6 of antibiotic administration, days 4-6 post-antibiotic administration, and days 14-16 post-antibiotic administration, were pooled for incubation, and chicks hatched from these pools of fertile eggs were treated with PREEMPT at hatch. When 48-h cecal propionate concentrations were used as an index of culture establishment, reduced (P < 0.05) efficacy was observed only in chicks derived from enrofloxacin-treated hens at either collection period. Although several antibiotics do not appear to produce detectable egg residues or interfere with CE culture establishment, these data suggest that chicks derived from enrofloxacin-treated hens may not be candidates for safe and effective CE culture treatment.

Animals↗

PubMed1978

Application of signal detection theory to error detection in ballistic motor skills.

Signal detection procedures were used to measure subjects' ability to detect errors in performance as they tried to hit a target in a ballistic motor task, since current methods were found to have several important shortcomings. Two tasks were used; one involved moving a slide 24.1 cm in 150 msec (temporal target), and the other involved rolling a ball at a visible target (spatial target). Subjects in both tasks were poor at detecting errors. Variability in the perceptual processes, measured by signal detection proceudres, was found to be about twice as much as that in the motor processes, measured by calculating variable error. This finding was inconsistent with a strictly closed-loop model of motor skills. Signal detection theory was also able to fill a gap in present theories of motor learning by explaining why some of the subjects in this experiment were able to detect errors better in the absence of knowledge of results.

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