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At least 757 records · Page 42Linked to original sources

Quantification of prostate shrinkage after microwave thermotherapy: a comparison of calculated cell-kill versus 3D transrectal ultrasound planimetry.

PURPOSE: To compare prostate shrinkage after transurethral microwave thermotherapy (TUMT) with calculated cell-kill. MATERIALS AND METHODS: The calculated cell-kill from 33 males with benign prostatic hyperplasia (BPH) treated with TUMT according to the ProstaLund Feedback Treatment (PLFT) method was compared to the post-treatment prostate volume change. The prostate volume was estimated with three-dimensional transrectal ultrasound (3D-TRUS) planimetry at baseline, 3, 6, and 12 months follow-up. A paired t-test was used to test the statistical significance of differences between the cell-kill volume and the prostate volume change. Linear regression was used to infer a relationship between the cell-kill and the 3D-TRUS data. The reproducibility of the 3D-TRUS method was assessed in repeated measurements. RESULTS: The mean prostate volume at baseline (N=33) was 56.1cm(3). After 3 (N=25), 6 (N=29) and 12 months (N=23), it was 45.5 cm(3), 39.7 cm(3), and 45.1cm(3), respectively. The corresponding average cell-kill volume was 16.4 cm(3), 17.1cm(3), and 17.2 cm(3), respectively. Predicted cell-kill volume was significantly larger than prostate shrinkage at 3 (p<0.0001), 6 (p=0.0002), and 12 months (p<0.0001), and showed a strong correlation at 3 and 6 months (r=0.74, p<0.0001). Correlation at 12 months was moderate (r=0.57, p=0.0041). Examination and investigation variability both averaged 2.5%. CONCLUSIONS: Cell-kill calculations of the PLFT method are proportional to the 3D-TRUS prostate shrinkage by a factor of 0.5 and have a precision of approximately +/-10 cm(3) for 90% of the patients during the first year after treatment.

Aged↗

Analysis of urinary 3-methoxy-4-hydroxyphenylglycol by high-performance liquid chromatography and electrochemical detection.

A high-performance liquid chromatographic method with electrochemical detection for the quantitation of total 3-methoxy-4-hydroxyphenylglycol (MHPG) in human urine is described. Existing methods for deconjugation and extraction have been optimized. The present method is simpler than existing methods with a high precision. Urinary MHPG is deconjugated enzymatically and subsequently extracted with ethyl acetate. The organic layer is extracted with acetic acid and a sample of the aqueous layer is injected into a reversed-phase column. In one run 90 samples can be processed. The critical parameters of deconjugation, extraction and chromatography are described. Data for reproducibility and selectivity are presented.

Chromatography, High Pressure Liquid↗

Capillary gas chromatographic assay for valproic acid and its 2-desaturated metabolite in brain and plasma.

A capillary gas chromatographic method has been developed for quantitating the antiepileptic drug valproic acid and its pharmacologically active metabolite, E-delta 2-valproic acid, in brain and plasma. The method was designed to cover large concentration ranges of both valproic acid and E-delta 2-valproic acid for pharmacokinetic and pharmacodynamic studies in laboratory animals and human subjects. Careful optimization of the extraction and chromatographic procedures was needed to resolve the analytes from a variety of endogenous constituents and other known metabolites of valproic acid. A sensitivity limit of 0.10 micrograms/g for 300 mg of brain tissue or 0.20 micrograms/ml for 150 microliters of plasma was achieved using flame ionization detection. The within-batch coefficients of variation for both analytes were less than 8%. Reproducible calibration data were observed over a period of three to eleven months.

Animals↗

Diffusion in thin films on the surface of a porous solid.

A water-wet mono-dispersed glass bead system is saturated with two phases, a wetting phase of water and a non-wetting phase of tetrachloroethylene (no 1H signal). Pulsed field gradient NMR measurements of the one-dimensional probability density distribution P(Delta) (X) for the diffusive displacements of water molecules in times, Delta, are presented for the whole accessible water saturation range. At lower water contents the distributions show a distinctive shape, which is attributed to the distribution of the aqueous phase in thin surface wetting films connecting pendular rings where the beads are in contact. The data are reproduced well by a computer simulation of a random walk model based on diffusion of molecules within such a structure, allowing determination of the surface film thickness.

Diffusion↗

Determination of isepamicin sulfate and related compounds by high performance liquid chromatography using evaporative light scattering detection.

A simple reversed phase high performance liquid chromatographic method was developed for the analysis of isepamicin sulfate. The use of evaporative light scattering detection eliminates the need for sample derivatization. Separation of the isepamicin aminoglycoside from structurally similar related compounds was achieved using two Waters X-Terra RP18 columns connected in tandem at 10 degrees C. The assay of isepamicin sulfate and the estimation of its impurities was accomplished using external standard calibration curves at two sample concentrations: 1.6 mg ml(-1) for the analysis of isepamicin sulfate and 8.0 mg ml(-1) for the estimation of lower level impurities. The limit of detection was 0.1%. The specificity, assay linearity, low level assay linearity and assay repeatability were also investigated.

Anti-Bacterial Agents↗

Fathead minnow (Pimephales promelas) vitellogenin: purification, characterization and quantitative immunoassay for the detection of estrogenic compounds.

The egg yolk precursor protein, vitellogenin (VTG), was purified from blood plasma of 17beta-estradiol (E2)-treated male fathead minnows (Pimephales promnelas) by anion-exchange chromatography on DEAE-agarose. A rabbit antiserum was raised against their blood plasma and then adsorbed with plasma from untreated (control) males to render the antiserum specific to VTG. The adsorbed antiserum was used to detect fathead minnow VTG (fVTG) in Western and dot blotting experiments and in an enzyme-linked immunosorbent assay (ELISA). The antiserum recognised fVTG as a approximately 156 kDa protein in plasma from vitellogenic females and E2-injected males but not untreated males. Its identity was confirmed by analysis of: (1) amino acid composition; (2) an internal amino acid sequence; (3) reactivity to the homologous antiserum; and (4) recognition by monoclonal antibodies prepared against the VTG from common carp (Cyprinus carpio) and brown bullhead (Ameiurus nebulosus). Specificity of the homologous antiserum to fVTG was confirmed by Western blotting of serially diluted plasma from vitellogenic females. Utility of the antiserum and purified fVTG for detecting exposure of male fathead minnows to estrogenic compounds was verified using a dot blotting immunoassay of fVTG and detected by chemiluminescence. Adult male fish were exposed to various concentrations of E2 (10(-8), 10(-9) and 10(-10) M) in their rearing water and plasma assayed for the presence of VTG at different time points (2, 7, 14 and 21 days). A competitive, antibody-capture, quantitative ELISA was then developed based on the purified fVTG and its respective antiserum. The ELISA was validated by demonstrating parallel binding slopes of dilution curves prepared with plasma from E2-injected males, vitellogenic females, and aqueous egg extracts as compared with purified fVTG standard. Plasma concentrations of VTG as low as 3 ng ml(-1) were detected in the ELISA, for which inter- and intra-assay coefficients of variation were both less than 5%. Furthermore, plasma from control males was unreactive with the fVTG antiserum. The VTG ELISA could be useful for the detection of estrogenic properties associated with certain compounds and could be easily incorporated into standard laboratory toxicity assays using this species.

Amino Acid Sequence↗

Good clinical practice, a transparent way of life. A review.

Investigators are regarded as central to the concept of clinical trials. Apart from bearing scientific responsibility, they will be confronted with a great number of ethical dilemma's concerning, amongst others, the justification and design of the trial, subject selection and informed consent, and presentation of the trial results. The ethical principles of biomedical research should be observed in accordance with a consistent moral code. The moral code of Good Clinical Practice (GCP) was developed in order to rule out inconsistencies. The principal tenets are that clinical trials should be good science, verifiable, monitored, well documented, and that they comply with the highest ethical standards. The principles of GCP are the protection of the rights of human subjects, integrity and reproducibility of data, and transparency of conduct.

Clinical Trials as Topic↗

Detection of high intensity transient signals (HITS): how and why?

HITS (high intensity transient signals) in transcranial Doppler recordings reflect either microemboli, both gaseous and solid, or artefacts. Various frequencies in number of microembolic signals (MES) have been reported in the same clinical condition. A possible explanation for these discrepancies may be different device settings and algorithms for embolus detection. For reproducibility of data, we suggest that studies on MES report the following parameters: (1) Ultrasound device; (2) transducer type; (3) insonated artery; (4) insonation depth; (5) algorithms for signal intensity measurement; (6) scale settings; (7) detection threshold; (8) axial extension of sample volume; (9) fast Fourier transform (FFT) size (number of points used); (10) FFT length (time); (11) FFT overlap; (12) transmitted ultrasound frequency; (13) high pass filter settings; and (14) recording time. No current system of automatic embolus detection has the full sensitivity and specificity required for clinical use. Therefore, each of the signals detected by these devices needs to be checked and verified by an experienced investigator. MES will help to identify the site and activity of the embolizing lesion. Microembolus detection might reduce the observation time and the number of patients needed to perform interventional trials. First, however, MES needs to be validated as a meaningful prognostic parameter. Microemboli originating from prosthetic cardiac valves are mainly gaseous. Therefore, they cannot serve as an indicator of the valves thromboembolic activity or the patient's stroke risk.

Carotid Stenosis↗

Long-chain polyamines (oligoamines) exhibit strong cytotoxicities against human prostate cancer cells.

alpha N,(omega)N-bis(ethyl) octamine SL-11160, decamine SL-11159, dodecamine SL-11226, and tetradecamine SL-11175 were chemically synthesized. We called this class of compounds 'oligoamines'. In these compounds, each -NH(2)(+) residue is separated by four CH(2) residues. trans-Unsaturation was also introduced into the center of the oligoamine chain resulting in the trans-octamine SL-11158, trans-decamine SL-11144, trans-dodecamine SL-11172 and trans-tetradecamine SL-11227. cis-Unsaturation gave the cis-octamine SL-11157 and cis-decamine SL-11150. When assayed for their growth inhibitory effect against four human prostate cancer cell lines LnCap, DU-145, DuPro, and PC-3 by a MTT assay, the ID(50) values were less than 1 microM in all four cell lines. On day 6 of treatment, 2 microM SL-11159, SL-11144 and SL-11175 killed over five logs of DuPro cells while SL-11172 killed over four logs as determined by a colony forming efficiency (CFE) assay. In addition, SL-11159, SL-11226 and SL-11227 killed four logs of PC-3 cells. PC-3 cells are generally resistant to shorter chain polyamine analogues. Such a level of cytotoxicity in any of the prostate tumor cell lines has not been observed for any other polyamine analogues tested thus far. The DU-145 cell line was too sensitive to oligoamines to perform a CFE analysis and the DuPro cell line was too sensitive to SL-11227 treatment to obtain reproducible CFE data. Interestingly, all 10 oligoamines were efficient DNA aggregators in a cell-free system and their cytotoxicities generally parallel their capacities to aggregate DNA.

Antineoplastic Agents↗

CEPH family 1362 STR database: an online resource for characterization of PCR products using electrospray ionization mass spectrometry.

An online database has been established in order to validate electrospray ionization mass spectrometry (ESI-MS) for genotyping and to publicize the procedures developed in our laboratory for the characterization of PCR products by ESI-MS. Genotypes derived from short tandem repeat (STR) loci that were obtained using ESI Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) have been posted for fifteen members of the CEPH family 1362 pedigree. The website provides specific information such as PCR parameters, PCR product cleanup approaches, and ESI solution compositions to enable other laboratories to reproduce our data. Links are provided to related websites in an effort to integrate information regarding the CEPH family, STR genotyping, and mass spectrometry. The database, currently available at http://www.people.vcu.edu/ -dcmuddim/genotype/ will be routinely updated with genotypes from additional STR loci including PCR parameters as well as PCR cleanup strategies as further developments are completed.

DNA, Complementary↗

Simultaneous analysis of 14 non-steroidal anti-inflammatory drugs in human serum by electrospray ionization-tandem mass spectrometry without chromatography.

A method is described for the simultaneous analysis of 14 non-steroidal anti-inflammatory drugs (NSAIDS) in human serum using negative electrospray ionization-tandem mass spectrometry (ESI-MS/MS). After addition of internal standard and protein precipitation using acetonitrile, samples were transferred to autosampler vials for direct analysis without chromatography. Injection of an air bubble with the sample and a multiple reaction monitoring (MRM) method using argon collision-induced dissociation (CID) of analyte (M-H)- ions permitted integration of the product ion peak areas to produce reproducible quantitative data over the range of concentrations expected in serum during routine use of these drugs. The method permitted the analysis of 30 samples per hour. Two hundred fifty consecutive analyses did not adversely affect instrument sensitivity.

Anti-Inflammatory Agents, Non-Steroidal↗

Inability to detect cell free fetal DNA in the urine of normal pregnant women nor in those affected by preeclampsia associated HELLP syndrome.

OBJECTIVE: Recent reports have indicated that cell-free fetal DNA can be detected in the urine of pregnant women. We attempted to reproduce those data. METHODS: Urine samples were collected from 18 normal pregnant women (11 with a male fetus). Urinary DNA was examined by Y-chromosome-specific nested polymerase chain reaction (PCR) or real-time PCR. Samples were also examined from two pregnancies complicated by HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome, which had very high levels of cell-free fetal DNA in the maternal plasma. To validate our data, a quantitative comparison of different DNA extraction procedures used in the previous reports was performed. RESULTS: In no instance were we able to detect any fetal DNA in maternal urine, although copious quantities of cell-free fetal DNA were present in the maternal plasma of those pregnancies affected by HELLP syndrome. Our quantitative analysis of the various extraction procedures used indicated that the commercial column elution method we used was comparable, if not superior, to the noncommercial methods used in previous reports. CONCLUSION: Our data strongly suggest that cell-free fetal DNA is not readily detectable in maternal urine, even under conditions known to increase kidney permeability.

Chromosomes, Human, Y↗

In vitro model to evaluate the relative efficacy of catheter-directed thrombolytic strategies.

RATIONALE AND OBJECTIVES: Catheter-directed thrombolytic therapy has become an accepted treatment for many vascular occlusions. However, the relative rates of lysis of the different methods of drug administration have not been quantified. We developed an in vitro model to simulate and quantify local lytic therapy of a thrombotic vascular occlusion and tested it by evaluating three catheter-directed lytic strategies. METHODS: Seven-centimeter-long segments of 125I-fibrinogen-labeled thrombus made from recently expired human blood from a blood bank were formed in plastic tubes and were placed in a flowing stream of saline. Using multisidehole catheters, the clots were "treated" with intrathrombic saline or urokinase administered by drip infusion or forced injection using identical total doses of drug and volumes of fluid. Using endhole catheters, saline or urokinase was drip infused into the leading edge of the thrombus using the same protocol. A collimated scintillation detector was used to quantify the amount of activity remaining in the thrombus during each experiment, and the resultant time-activity curves for the different trials were compared. RESULTS: Forced-injection administration of urokinase using a multisidehole catheter produced the fastest lysis, resulting in a half-life of 42 min. The other infusion methods were slower, with half-lives of 153 min for multisidehole urokinase drip infusion, 365 min for endhole urokinase drip infusion, and more than 1,000 min for multisidehole catheter forced injection of saline and multisidehole and endhole saline drip infusion. The differences among these groups were reproducible and statistically significant. CONCLUSION: Results suggest that a simple and inexpensive in vitro model simulating lysis of a vascular occlusion can produce reproducible quantitative data. The data demonstrate that forced injection of lytic agents with a multisidehole catheter enhances the rate of thrombolysis and that the enhancement is not primarily attributable to the mechanical effect of this mode of administration.

Catheterization, Peripheral↗

Contact lens standards.

The British Standards Institution (BSI) is in the process of publishing an extensive series of standards relating to contact lenses which were originally formulated as International and European Standards. Standards specifying tolerance limits for the manufacture of rigid and hydrogel lenses have been based upon data on reproducibility derived from 'ring tests' in which measurements are made by a series of independent operators. Despite publication of an International Standard on terminology in 1986, some authors persist in using obsolete terms and this practice is deplored.

Journal Article↗

New chips for molecular biology and diagnostics.

The DNA chips are arrays of DNA probes immobilized on solid support for simultaneous identification of many target DNA sequences. DNA chips applied to diagnosis aims to detect genomic DNA or RNA after PCR amplification. This review provides an overview of DNA chip technology, focusing on diagnostic applications. A comparison between high density and low density microarrays is given showing that low density chips are more suitable for routine applications due to their simplicity, good reproducibility, easy data management and low cost.

Bacterial Infections↗

Identification of two proteins induced by exposure of the pathogenic fungus Candida glabrata to fluconazole.

Candida glabrata is an increasingly important cause of opportunistic fungal infection of humans and appears to be intrinsically resistant to the triazole antifungal fluconazole. However, the mechanisms responsible for reduced susceptibility to azole drugs are not understood. Fluconazole exposure rapidly induced expression of a 169-kDa protein band in plasma membrane fractions of C. glabrata cells. Mass spectrometry of trypsin-digested peptide fragments showed that the induced protein band comprised the ATP binding cassette-type drug efflux transporter CgCdr1p. CgCdr1p was also functionally overexpressed in S. cerevisiae and similarly identified by mass spectrometry. A 61-kDa protein band in the plasma membrane fraction from C. glabrata was also induced by fluconazole exposure. Mass spectrometric peptide fingerprinting identified this band as lanosterol 14alpha-demethylase, the enzyme in the ergosterol biosynthesis pathway targeted by fluconazole. The rapid induction of a multidrug efflux pump and/or overproduction of lanosterol 14alpha-demethylase are mechanisms that could make C. glabrata appear intrinsically resistant to fluconazole. Mass spectrometric fingerprint analysis of SDS-PAGE separated plasma membrane fractions combined with heterologous hyper-expression provides a convenient method for protein identification and functional evaluation of induced proteins, even in an organism where the genome sequence database is incomplete.

Amino Acid Sequence↗

Selective detection of chlorine-containing compounds by gas chromatography/chemical reaction interface mass spectrometry.

Chemical reaction interface mass spectrometry (CRIMS) was studied as a gas chromatographic detection technique for chlorine-containing compounds. Both SO2 and HBr were tested as reactant gases. With SO2, a detection limit of 50 pg of diazepam and a linear range of 4 orders of magnitude were achieved, and the experimental data were reproducible. With HBr, the detection limit was 10 ng of diazepam and the linear dynamic range was only 2 orders of magnitude. The possible pharmacological application of CRIMS was studied using urine spiked with diazepam and several of its metabolites, and the results show CRIMS to be a simple but potentially powerful method in drug metabolism studies.

Acids↗

Electrooxidation of ascorbic acid on a polyaniline-deposited nickel electrode: surface modification of a non-platinum metal for an electrooxidative analysis.

The electrooxidation of ascorbic acid (H2A), which does not occur on a bare Ni electrode, has been shown to take place on a polyaniline (PANI)-coated Ni electrode in aqueous electrolytes of a wide pH range. The characteristic voltammetric peak of PANI in 0.1 M H2SO4 at 0.2 V vs SCE corresponding to the transformation of leucoemeraldine to emeraldine gradually diminishes with an increase in concentration of H2A as a result of adsorption. This peak disappears before the appearance of another peak corresponding to the oxidation of H2A at a concentration of 1 mM. The irreversible oxidation current of H2A exhibits a linear dependence on the concentration. The effect of adsorption of H2A on PANI has been shown to increase the voltammetric peak current. A study on the variation of the PANI thickness and its influence on the voltammetric oxidation of H2A has led to an optimum thickness of 1.6 microm. The oxidation currents on the porous PANI/Ni electrode have been found to be several times higher at lower potentials in comparison with the data of a Pt electrode. The reaction has also been studied by ac impedance spectroscopy. In alkaline electrolytes, the Nyquist impedance plot is characterized by two semicircles instead of a single semicircle in acidic electrolytes. Thus, Ni, which is a non-platinum metal, has been found to be useful, by surface modification with PANI, for electrooxidation of H2A. The data are reproducible in the electrolytes of a wide pH range, thus suggesting a good stability, reusability and a long life for the PANI/Ni electrodes.

Adsorption↗