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At least 1,531 records · Page 85Linked to original sources

A test for the identification of relevant sympathetic nerve fibers during nerve sparing retroperitoneal lymphadenectomy.

We present a new intraoperative test for the identification of sympathetic nerve fibers relevant to ejaculation during nerve sparing retroperitoneal lymphadenectomy in patients with nonseminomatous testicular tumors. Electrostimulation of specific isolated postganglionic nerve fibers resulted in intraoperative ejaculation in 9 of 11 patients. Ejaculation was without tumescence in a noneruptive manner and was reproducible in most cases upon repeated stimulation. In 6 patients ejaculation resulted after stimulation of the L3 fiber and in 1 each after stimulation of the L1, L2 and hypogastric plexus. The test seems to be particularly useful in post-chemotherapeutic dissections or otherwise extended dissections when sparing of irrelevant fibers is to be avoided.

Adrenergic Fibers↗

Is the loss of endothelial thrombomodulin involved in the mechanism of chronicity in late radiation enteropathy?

BACKGROUND AND PURPOSE: Radiation enteropathy is characterized by locally elevated levels of inflammatory and fibrogenic cytokines. Microvascular injury may sustain these alterations through persistent local hypercoagulopathy, platelet aggregation, leukocyte adhesion and release of biologically active mediators. This study assessed the relationship of endothelial thrombomodulin (TM), a key regulator of the protein C anticoagulant pathway and marker of endothelial function, with transforming growth factor beta (TGF-beta) immunoreactivity and morphologic alterations in radiation enteropathy. MATERIALS AND METHODS: Small bowel resection specimens from 9 patients with radiation enteropathy were analyzed by computerized quantitative immunohistochemistry using antibodies against TM, von Willebrand factor (vWF) and TGF-beta. Identical measurements were performed on intestinal resection specimens from otherwise healthy penetrating trauma victims and on archived small intestines. A previously validated image analysis technique was used to assess submucosal vessels for TM and vWF immunoreactivity, and the intestinal wall for total extracellular matrix-associated TGF-beta immunoreactivity. RESULTS: Specimens from irradiated patients showed prominent submucosal and subserosal thickening and fibrosis, and obliterative vasculopathy. Control specimens were histopathologically normal. Vascular density and vWF immunoreactivity were similar in radiation enteropathy patients and controls. The image-analysis techniques were highly reproducible, with correlation coefficients for repeated measurements ranging from 0.86 to 0.93. Radiation enteropathy specimens exhibited a highly significant reduction in the number and proportion of TM-positive submucosal vessels per unit area (P < 0.0001) and increased intestinal wall TGF-beta immunoreactivity (P = 0.002). CONCLUSIONS: These data support the theory that sustained endothelial dysfunction is involved in the molecular pathogenesis of radiation enteropathy, and point to TM as important in the chronic nature of radiation enteropathy and a potential target for prophylactic and therapeutic interventions.

Aged↗

A comparison of 3-D data correlation methods for fractionated stereotactic radiotherapy.

PURPOSE: Stereotactic radiosurgery is currently used to treat patients who are not good candidates for conventional neurosurgical procedures. For treatments of nonvascular tumor cells, it appears that fractionation offers a radiobiological advantage between tumor and normal tissues. Therefore, fractionated stereotactic radiotherapy (FSR) is preferred because it minimizes normal tissue complications and maximizes local tumor control probability. We have implemented a methodology clinically to perform the noninvasive patient repositioning technique. The 3-D data correlation method for high-precision and multiple fraction stereotactic treatments has been presented. METHODS AND MATERIALS: Three different optimization algorithms (Hooke and Jeeves optimization, simplex optimization, and simulated annealing optimization) are evaluated to calculate the transformation parameters necessary for FSR. A least-square object function is created to perform the 3-D data matching process. By minimizing the unconstrained object function value the best fit can be approached for the reference 3-D data sets. Simulation shows that these algorithms deliver results that are comparable to the previously published correlation algorithm (1,2) (singular value decomposition [SVD] method). The advantage for optimization algorithms is easily understood and can be readily implemented by using a personal computer (PC). The mathematical framework provides a tool to calculate the transformation matrix which can be used to adjust patient position for fractionated treatments. Therefore, using these algorithms for a high-precision fractionated treatment is possible without an invasive repeat fixation device and has been implemented clinically. A bite plate system was incorporated to acquire 3-D patient data. With a 3-D digital camera localization device, the patient motion can be followed in real time with the system calibrated to the isocenter. RESULTS: Two types of data sets are utilized to study the correlation results. One is using the digitized patient data which were retrieved clinically. The other is using the randomly generated data sets. Simulation errors for the optimization algorithms are all less than 1 mm in translation and less than 1 degree in rotation. Currently, FSR is performed using special designed repeat fixation devices which assure reproducible patient position for multiple fractions of radiation treatment. Clinical results indicated that this technique provided excellent treatment results. CONCLUSION: Three optimization algorithms have been applied and evaluated in calculating the transformation parameters between two 3-D contours or digitized data points. The mathematical functions behind these optimization algorithms are straightforward and can be easily implemented. When incorporated with the proper CT/MR image data with an electronic portal imaging (EPI) system, this process can possibly verify the patient's treatment position whenever there is doubt about the movement during the treatment procedure.

Algorithms↗

Kinase receptor activation (KIRA): a rapid and accurate alternative to end-point bioassays.

We have developed a novel strategy for a rapid bioassay that is accurate, precise, sensitive, and high capacity. It is capable of quantifying ligand bioactivity by measuring ligand-induced receptor tyrosine kinase activation in terms of receptor phosphorylation. The assay, termed a 'kinase receptor activation' or KIRA, utilizes two separate microtiter plates, one for ligand stimulation of intact cells, and the other for receptor capture and phosphotyrosine ELISA. The assay makes use of either endogenously expressed receptors or stably transfected receptors with a polypeptide flag. KIRA assays for the ligands IGF-I and NGF were compared to their corresponding endpoint bioassays (3T3 cell proliferation for IGF-I and PC12 cell survival for NGF). The KIRA assays showed excellent correlation with the more classical endpoint bioassays. Further, they were highly reproducible, minimizing the requirement for repeat assays. The KIRA assay format has great potential as a rapid, accurate and precise bioassay, both for potency determination as well as stability-indicating analyses.

Biological Assay↗

Comparison of derivative spectrophotometric and liquid chromatographic methods for the determination of omeprazole in aqueous solutions during stability studies.

A first derivative spectrophotometric method was developed for the determination of omeprazole in aqueous solutions during stability studies. The derivative procedure was based on the linear relationship between the omeprazole concentration and the first derivative amplitude at 313 nm. The first derivative spectra was developed between 200 and 400 nm (deltalambda = 8). This method was validated and compared with the official high-performance liquid chromatography (HPLC) method of the USP. It showed good linearity in the range of concentrations studied (10-30 microg ml(-1)), precision (repeatability and inter-day reproducibility), recovery and specificity in stability studies. It also seemed to be 2.59 times more sensitive than the HPLC method. These results allowed to consider this procedure as useful for the rapid analysis of omeprazole in stability studies since there was no interference with its decomposition products.

Anti-Ulcer Agents↗

Cellular electrophysiologic changes and "arrhythmias" during experimental ischemia and reperfusion in isolated cat ventricular myocardium.

The cellular electrophysiologic consequences of both regional and global experimental ischemia and reperfusion were studied in the isolated cat myocardium, using conventional microelectrode techniques. Oxygenated Tyrode's solution was perfused through the left anterior descending and circumflex coronary arteries, while the preparation was superfused with Tyrode's solution gassed with 95% nitrogen and 5% carbon dioxide. Electrophysiologic characteristics of endocardial muscle cells were normal during coronary perfusion. When perfusion was discontinued for 30 minutes, resting membrane potential was decreased by 21.6 +/- 4.1%, action potential amplitude was decreased by 29.1 +/- 8.6% and action potential duration was decreased by 54.1 +/- 12.5% (p less than 0.001). Ectopic activity occurred after 5 to 10 minutes of ischemia and was more frequent in regional than in global ischemia (p less than 0.05). Rapid ventricular activity was observed in only 5 (17%) of 29 preparations during ischemia, whereas it occurred in 24 (83%) of 29 preparations during reperfusion. Rapid ventricular activity began 5 to 40 seconds (mean 19) after the start of reperfusion, stopped spontaneously after a mean of 113 +/- 211 seconds and occurred after both regional and global ischemia. The cellular electrophysiologic changes induced by ischemia returned to baseline values within the next 5 minutes. Repeated ischemia and reperfusion runs reproduced the same electrophysiologic changes and rapid ventricular activity. Coronary perfusion with procainamide (20 mg/liter) aggravated the ischemic depressions of action potential amplitude and action potential duration and increased conduction delay during ischemia, but it did not prevent rapid ventricular activity induced by reperfusion. In contrast, verapamil (1 mg/liter) perfusion did not affect the changes in action potential variables during ischemia but prevented reperfusion-induced rapid ventricular activity. Perfusion with calcium ion (Ca2+)-free Tyrode's solution just before ischemia and during reperfusion slowed or prevented reperfusion-induced rapid ventricular activity, without affecting the action potential changes during ischemia. It is concluded that, in these isolated perfused ventricular muscle preparations, different mechanisms may be operative in ischemic and reperfusion arrhythmias and Ca2+ may play an important role in the development of arrhythmias during the reperfusion phase of ischemia/reperfusion sequences.

Action Potentials↗

Amperometric immunosensor for lactate dehydrogenase LD-1.

An amperometric immunosensor for the detection of the lactate dehydrogenase isoenzyme LD-1 has been developed. Polyclonal antibodies for LD-1 have been covalently immobilised onto a preactivated Immunodyne ABC membrane, reacted with standard LD-1 solutions and placed onto a platinum working electrode polarised at +600 mV vs. Ag/AgCl. Lactate dehydrogenase activity has been measured by detection of the oxidation of NADH at the electrode surface. A calibration curve for LDH in the 0.005-0.12 U range has been obtained with a repeatability of 3% and a reproducibility of 10%. The probe exhibited good selectivity (response of LD-2 was 18% of LD-1) which was further improved using thermal treatment of the membranes. Measurement of LD-1 content in human control serum with the developed procedure gave a LD-1 concentration in the reported assigned range.

Animals↗

Measurements of gallbladder motor function by ultrasonography: towards standardization.

As real-time ultrasonography is a cheap, noninvasive, relatively easy, validated and reproducible technique, it can be repeated over time to document time-related changes of gallbladder motor function. Ultimately, functional ultrasonography estimates gallbladder shape and volume in fasting state and in response to a test meal (liquid or mixed solid-liquid, provided there is sufficient fat content) or exogenous stimulus (e.g., i.v. cholecystokinin or ceruletide). Although functional ultrasonography of the gallbladder has been mainly used for research purposes in specific referral centres, its simplicity makes such a technique appealing in the clinical setting to assess gallbladder motor function in both health and disease. Indications include the study of healthy subjects and of patients during pathophysiologically relevant conditions; in particular when subjects are at risk for gallbladder stasis and gallstone disease or during gallstone disease when a decision concerning medical dissolution therapy is required.

Cholecystolithiasis↗

Liquid chromatography at the critical condition for polyisoprene using a single solvent.

Liquid chromatography at the chromatographic critical condition has drawn much attention as an attractive characterization method of block copolymers since it has been proposed that a part of a polymer chain becomes "chromatographically invisible" at this condition, which would permit the characterization of individual blocks. A critical condition for a polymer species has been commonly established by use of mixed-solvent systems. It is not easy, however, to reproduce the critical condition since the retention of polymers depends very sensitively on the solvent composition and purity. Furthermore, the preferential sorption of a component in a mixed solvent may cause an additional problem. Therefore, the use of a single solvent is highly desirable to improve the reproducibility as well as the repeatability. In this study, a single-solvent critical condition for polyisoprene was established with 1,4-dioxane and C18 bonded silica as the mobile and stationary phases, respectively. At this condition, the "chromatographic invisibility" of polystyrene-polyisoprene diblock copolymers was critically examined and it was found that a rigorous chromatographic invisibility was not achieved and the retention of the block copolymers was affected by the length of the blocks under the critical condition. Some other chromatographic applications using the single-solvent system are also reported.

Journal Article↗

Evaluation of polyphenolic and flavonoid compounds in honeybee-collected pollen produced in Spain.

Polyphenolic content, flavonoid content, and free flavonoid aglycon compounds were determined in 11 samples of Spanish honeybee-collected pollen. Adequate extraction was obtained with ethyl acetate in the determination of free flavonoid aglycon. Recovery (>83.6%), within-run repeatability (<6.67%), between-run reproducibility (<8.73%), and detection limits (1.4--1.9 mg/kg) were satisfactory. A total of 15 compounds were separated with gradient reversed phase HPLC, and 13 were identified and quantified using diode array detector. The most predominant compounds were flavonoid glycosides, mainly flavonols. Eighty-two percent of the samples contained at least 14 of the phenolic components, primarily rutin, quercetin, myricetin, and trans-cinnamic acid as free aglycons. Total phenols were present, at levels of >0.85 g/100 g in the form of non-tannins, and flavonoids of >0.35 g/100 g, using spectrophotometric procedures. Rutin is the best identifier of free flavonoid aglycon compounds. A minimum quantity of 200 mg/kg of rutin is suggested to guarantee the nutritional and biological properties required in the European market.

Animals↗

Inverse gas chromatographic method for measurement of interactions between soy protein isolate and selected flavor compounds under controlled relative humidity.

An inverse gas chromatographic (IGC) method was developed to study the binding interactions between selected volatile flavor compounds and soy protein isolate (SPI) under controlled relative humidity (RH). Three volatile probes (hexane, 1-hexanol, and hexanal) at very low levels were used to evaluate and validate system performance. On the basis of the thermodynamic data and the isotherms measured at 0% RH, 1-hexanol and hexanal had higher binding affinities than hexane, which could be attributed to hydrogen-bonding interactions with SPI. At 30% RH, 1-hexanol and hexanal were retained less than at 0% RH, indicating possible competition for binding sites on the SPI surface between water and volatile probe molecules. Results showed that the thermodynamic data determined were comparable to the available literature values. Use of IGC allowed for the rapid and precise generation of sorption isotherms. Repeatability between replicate injections and reproducibility across columns were very good. IGC is a potentially high-throughput method for the sensitive, precise, and accurate measurement of flavor-ingredient interactions in low-moisture food systems.

Adsorption↗

Development of a fast analytical method for the determination of sudan dyes in chili- and curry-containing foodstuffs by high-performance liquid chromatography-photodiode array detection.

A simple and fast analytical method for the determination of sudans I, II, III, and IV in chili- and curry-containing foodstuffs is described. These dyes are extracted from the samples with acetonitrile and analyzed by high-performance liquid chromatography coupled to a photodiode array detector. The chromatographic separation is carried out on a reverse phase C18 column with an isocratic mode using a mixture of acetonitrile and water. An "in-house" validation was achieved in chili- and curry-based sauces and powdered spices. Depending on the dye, limits of detection range from 0.2 to 0.5 mg/kg in sauces and from 1.5 to 2 mg/kg in spices. Limits of quantification are between 0.4 and 1 mg/kg in sauces and between 3 and 4 mg/kg in spices. Validation data show a good repeatability and within-lab reproducibility with relative standard deviations < 15%. The overall recoveries are in the range of 51-86% in sauces and in the range of 89-100% in powdered spices depending on the dye involved. Calibration curves are linear in the 0-5 mg/kg range for sauces and in the 0-20 mg/kg range for spices. The proposed method is specific and selective, allowing the analysis of over 20 samples per working day.

Azo Compounds↗

Pressurized liquid extraction of capsaicinoids from peppers.

A method has been developed for the extraction of capsaicinoids from peppers by pressurized liquid extraction (PLE); these compounds are determined by reverse phase high-performance liquid chromatography (HPLC), with detection by fluorescence spectrophotometry and mass spectrometry (MS). The stability of capsaicin and dihydrocapsaicin has been studied at different temperatures (50-200 degrees C), and several extraction variables have been assayed: solvent (methanol, ethanol, and water), different percentages of water in the methanol (0-20%) and in the ethanol (0-20%), and the number of extraction cycles. The study has evaluated the repeatability (RSD < 7%) and the reproducibility (RSD < 7%) of the method. Finally, the PLE method developed has been applied to quantify the capsaicinoids present in three varieties of hot peppers cultivated in Spain, quantifying five capsaicinoids: nordihydrocapsaicin, capsaicin, dihydrocapsaicin, an isomer of dihydrocapsaicin, and homodihydrocapsaicin.

Capsaicin↗

Development of a micro-fluidic manifold for copper monitoring utilising chemiluminescence detection.

The progressive development of a micro-fluidic manifold for the chemiluminescent detection of copper in water samples, based on the measurement of light emitted from the Cu(ii) catalysed oxidation of 1,10-phenanthroline by hydrogen peroxide, is reported. Micro-fluidic manifolds were designed and manufactured from polymethylmethacrylate (PMMA) using three micro-fabrication techniques, namely hot embossing, laser ablation and direct micro-milling. The final laser ablated design incorporated a reagent mixing channel of dimensions 7.3 cm in length and 250 x 250 microm in width and depth (triangular cross section), and a detection channel of 2.1 cm in length and 250 x 250 microm in width and depth (total approx. volume of between 16 to 22 microL). Optimised reagents conditions were found to be 0.07 mM 1,10-phenanthroline, containing 0.10 M cetyltrimethylammonium bromide and 0.075 M sodium hydroxide (reagent 1 delivered at 0.025 mL min(-1)) and 5% hydrogen peroxide (reagent 2 delivered at 0.025 mL min(-1)). The sample stream was mixed with reagent 1 in the mixing channel and subsequently mixed with reagent 2 at the start of the detection channel. The laser ablated manifold was found to give a linear response (R(2) = 0.998) over the concentration ranges 0-150 microg L(-1) and be reproducible (% RSD = 3.4 for five repeat injections of a 75 microg L(-1) std). Detection limits for Cu(ii) were found to be 20 microg L(-1). Selectivity was investigated using a copper selective mini-chelating column, which showed common cations found in drinking waters did not cause interference with the detection of Cu(ii). Finally the optimised system was successfully used for trace Cu(ii) determinations in a standard reference freshwater sample (SRM 1640).

Catalysis↗

Impact of electrocardiogram recording format on QT interval measurement and QT dispersion assessment.

The aim of this study was to determine the effect of recording conditions on the operator dependent measures of QT dispersion in patients with known and/or suspected repolarization abnormalities. Among several methods for risk stratification, QT dispersion has been suggested as a simple estimate of repolarization abnormalities. In a cohort of high and low risk patients, different components of the repolarization process were assessed in the 12-lead ECG using three different paper speeds and amplifier gains. To assess measurement error and reproducibility, a straight line was repeatedly measured. The operator error was 0.675 +/- 0.02 mm and the repeatability of the measurement error was 31 +/- 6%. The QT interval was most frequently measurable in V2-V5. Depending on the lead selected for analysis, the incidence of visible U waves was greatest in the precordial leads with high amplifier gain and low paper speed, strongly affecting QT interval measurement. The timing of the onset of the QRS complex (QRS onset dispersion) or offset of the T wave was strongly dependent on the paper speed. Paper speed, but not amplifier gain, had a significant shortening effect on the measurement of the maximum QT interval. As QT interval measurement in each ECG lead incorporates QRS onset and T wave offset (depending on the number of visible U waves), the dispersion of each of these parameters significantly affected QT dispersion. Thus, QT dispersion appears to reflect merely the presence of more complex repolarization patterns in patients at risk of arrhythmias.

Adult↗

Validation of the measurement of glucose appearance rate with [6,6-2H2]glucose in lactating dairy cows.

The aim of this study was to validate the measurement of glucose appearance rate using [6,6-2H2]glucose i.v. infusion in lactating dairy cows. Sample enrichments were analysed by gas chromatography/mass spectrometry. Linearity (enriched solutions) and specificity (enriched plasma) were good: for enrichments ranging between 1.6 and 6.3 mol% excess, the slopes were about 1 and the ordinates at the origin were not different from zero. For a plasma enriched at 3.74 mol% excess, repeatability and long term intralaboratory reproducibility coefficients of variation were 1.31 and 1.90%, respectively. The appearance rates were calculated by two models. The values provided by the steady-state model were not different from those provided by the non-steady-state Steele model. Both models can be used because the treatment effects were similarly discriminated regardless of the model. In our experiments analysing the nutritional effects on Ra in mid-lactating cows, the precision of the method (1.90%) was not the limiting factor to detect a significant difference in Ra compared to the statistical precision obtained with the experimental scheme (4 x 4 and 5 x 5 Latin square design). We conclude that in lactating dairy cows, the measurement of glucose fluxes with this method is relevant and minimally invasive for the animals.

Animals↗

Intra- and inter-observer variability of basal flow velocity and vascular reactivity measurements using transcranial Doppler sonography.

The reliability of blood flow velocity measurements performed using transcranial Doppler sonography was investigated with regard to the intra- and inter-observer variability. We compared the flow velocity values recorded in the intracranial vessels by two experienced investigators both under rest conditions and during hypercapnia induced by an apnea test. The results showed how flow velocity values measured at the level of the middle cerebral artery, anterior cerebral artery, ophthalmic artery and basilar artery were not significantly influenced by intra- or inter-examiner variability. The flow velocity increases recorded from the middle cerebral artery during the activation test proved to be reproducible when the measurements were repeated by the same examiner, but more variable when detected by different investigators. On the basis of the available data, the clinical relevance of transcranial Doppler results must take into account the degree of reliability of its measurements.

Apnea↗

Analysis of chloramphenicol in honeys of different geographical origin by liquid chromatography coupled to electrospray ionization tandem mass spectrometry.

A sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to detect trace amounts of the antibiotic chloramphenicol (CAP) in honey. The methodology entailed a solid-phase extraction of aqueous honey solutions followed by liquid-liquid partitioning, filtration and direct injection onto the LC-MS/MS system. Honey extracts were spiked with an isotopically labelled internal standard (d(5)-CAP) to compensate for analyte loss and potential ion suppression during the MS stage. Detection of the analyte was achieved by negative ionization electrospray in the selected reaction monitoring (SAM) mode. For confirmation, four characteristic mass transitions were monitored each for the analyte and the surrogate standard. The method was validated according to the latest European Union criteria for the analyses of veterinary drug residues in food. At all three fortification levels studied (0.1, 0.2, 0.5 microg kg(-1)) the method was accurate to within 15%. The repeatability and within-laboratory reproducibilities were <12 and 18%, respectively. The decision limit (CC alpha) and detection capability (CC beta) were both <0.1 microg kg(-1). The procedure provides a sensitive and reliable method for the determination of residues of chloramphenicol in honey. Numerous raw honeys of various geographical origins were analysed, showing extensive contamination particularly those of Chinese origin.

Animals↗