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Effect of pattern of aerosol inhalation on clearance of technetium-99m-labeled diethylenetriamine pentaacetic acid from the lungs of normal humans.

The clearance rate of inhaled aerosols of technetium-99m-labeled diethylenetriamine pentaacetic acid (99mTc-DTPA) from the lungs provides a rapid, clinically useful, noninvasive index of pulmonary epithelial permeability. In order to identify a method that minimizes intrasubject and intersubject variability and thereby provides a reliable means to identify patients with abnormal values, we administered a submicronic aerosol of 99mTc-DTPA to 10 healthy, nonsmoking male subjects with either tidal breathing (Vtidal) or multiple vital capacity maneuvers (VVC). Subjects then spontaneously breathed room air while counting continued for 30 min. Monoexponential clearance rates over 7, 15, and 30 min were compared with a two-compartment, biexponential analysis over 30 min. Intrasubject reproducibility was evaluated by repeating clearance 2 to 156 days later. Monoexponential clearance following VVC at 30 min equaled 1.36 +/- 0.55%/min compared with 0.83 +/- 0.25%/min for Vtidal (p less than 0.025). VVC inhalations resulted in a larger fast compartment of 16 +/- 12% compared with 3 +/- 2% with tidal breathing (p less than 0.01). The least intrasubject variability with coefficient of variation (CV) of +/- 18% was obtained with monoexponential analyses after Vtidal during 15 min of scanning and with either breathing maneuver over 30 min. Monoexponential clearance for 30 min with Vtidal gave the least scatter between subjects, with CV of +/- 30%. These data show that simple tidal inhalations of 99mTc-DTPA followed by a monoexponential analysis of the 30-min time-activity curve from both lungs minimize the degree of variability between and among subjects and provide a predicted normal value of clearance of 0.83 +/- 0.25%/min. The development of a more rapid curvilinear clearance followed by delivery VVC suggests that several deep breaths transiently increase epithelial permeability or reduce the volume of liquid in the alveolar subphase in some regions. Resting for 20 min prior to inhaling the aerosol of 99mTc-DTPA is recommended to avoid alterations in clearance rates from deep breathing.

Administration, Inhalation↗

Cartilage volume quantification via Live Wire segmentation.

RATIONALE AND OBJECTIVES: A reduction in cartilage volume is characteristic of osteoarthritis and hence there exists a need for an accurate and reproducible method to measure in vivo cartilage volume. Quantification of cartilage volume from magnetic resonance (MR) images requires a segmentation technique such as the user-driven "Live Wire" strategy that can reliably delineate object volumes in a time-efficient manner. In the present work, the accuracy and reproducibility of the Live Wire method for the quantification of cartilage volume in MR images is evaluated. MATERIALS AND METHODS: The accuracy of the Live Wire method was assessed by comparing the MR-based volume measurement of a patellar cartilage-shaped phantom versus data calculated via water displacement. The inter- and intra-operator reproducibility of the technique was evaluated from Live Wire segmentation of the patellar cartilage volume from fat-suppressed 3-dimensional spoiled-gradient-echo images of five healthy human volunteers performed by three operators. To provide data for analysis of inter-scan reproducibility, the human scans were repeated five times with the aid of a leg-restraining jig to minimize repositioning error. RESULTS: The volume of the patellar cartilage-shaped phantom measured via Live Wire segmentation of MR images was within 97.8% of its true volume. The average inter- and intra-operator coefficients of variation of three operators were 3.0% and 0.4%, respectively. The average inter-scan coefficient of variation of five repeated scans of each volunteer was 2.7%. CONCLUSION: The data suggest that the Live Wire strategy is an accurate, reproducible, and efficient technique to measure cartilage volume in vivo in a feasible amount of operator time.

Adult↗

The reproducibility of endometrial volume acquisition and measurement with the VOCAL-imaging program.

OBJECTIVES: To determine the intraobserver reproducibility of volume acquisition and repeatability of endometrial volume measurements using the VOCAL-imaging program (Virtual Organ Computer-aided AnaLysis). METHODS: Ten three-dimensional (3D) ultrasound datasets of the same endometrium were obtained from one patient during one scan session. For every 3D dataset, the endometrial volume was calculated by four different measurement techniques; three rotational methods and one conventional method. A single observer acquired each dataset and conducted all measurements. For the rotational technique, the manual mode was used to trace the myometrial-endometrial border through three different predefined rotations of 30 degrees, 15 degrees and 9 degrees . Endometrial volume was also measured in a conventional manner by scrolling through one of the multiplanar images and outlining the contour in another. Ten consecutive measurements were taken for each of the four measurement techniques so that 40 endometrial volume readings were obtained for each of the 10 3D-ultrasound datasets. RESULTS: Endometrial volume measurements were more reproducible with the rotational technique. Rotational methods employing a rotation step of less than 30 degrees were associated with coefficients of variation below 1% and intraclass correlation coefficients above 0.94. Within each of the 10 different scans, the homogeneity of the variance of measurements was significantly different according to the method of measurement (P = 0.022 to < 0.001). Post-hoc tests revealed that both the conventional and 30 degrees rotational methods resulted in a significantly smaller mean endometrial volume than both the 15 degrees and the 9 degrees rotational methods. CONCLUSIONS: Endometrial volume acquisition is reproducible and endometrial volume measurements are repeatable but they depend upon the technique of volume calculation in so much as rotational methods employing a rotation step of less than 30 degrees are associated with a significantly smaller variance in measurements and a significantly greater mean endometrial volume.

Adult↗

Prevalence and reproducibility of exercise-induced ventricular arrhythmias during maximal exercise testing in normal men.

The occurrence of ventricular arrhythmias at rest or during ordinary daily activities has been implicated as a risk factor for future coronary-related events and sudden death. However, the clerical significance of exercise-induced ventricular arrhythmias remains uncertain. To assess the prevalence and reproducibility of such arrhythmias, two serial maximal treadmill exercise tests were performed in a study population of 543 male Indian State policemen at an average interval of 2.9 years. Four hundred sixty-two subjects were clinically free of evidence of cardiovascular disease, and 81 had evidence of definite or suspected cardiovascular disease. The prevalence of exercise-induced ventricular arrhythmias during the first test was 30% in men aged 25 to 34 years, 32% in those aged 35 to 44 years and 36% in those aged 45 to 54 years. The prevalence rate in these age groups with repeat testing was 36, 38 and 42%, respectively. These differences were not statistically significant. The group with definite or suspected cardiovascular disease had a greater prevalence of exercise-induced ventricular arrhythmias than normal subjects during both tests but the prevalence rate with repeat testing remained constant. The occurrence of exercise-induced ventricular arrhythmias was reproducible in individual subjects during the second test in 55% of 25 to 34 year olds, 58% of 35 to 44 year olds and 62% of 45 to 54 year olds. Thus, individual reproducibility in two consecutive tests was only slightly greater than reproducibility by chance alone. The group with known or suspected cardiovascular disease demonstrated a trend toward greater reproducibility with repeat testing. Exercise-induced ventricular arrhythmias were not reproducible by type or complexity. The marked variability of exercise-induced ventricular arrhythmias during repeat maximal exercise testing in a clinically normal population appears to negate the usefulness of this finding during a single test as a marker of future cardiovascular disease. Nevertheless, subjects whose arrhythmias were reproducible may form a group destined to manifest clinical cardiovascular disease in long-term follow-up studies.

Adult↗

Perfusion-corrected mapping of cardiac regional blood volume in rats in vivo.

Measurement of regional blood volume (RBV) in the myocardium in vivo is important for the assessment of tissue viability and function. The method in this work is based on the acquisition of a T(1) map before and after intravascular contrast agent application. It is known that this method is influenced by perfusion that causes an overestimation of RBV values. In order to solve this problem, the new method is proposed which acquires T(1) maps with slice selective inversion pulses. Due to blood flow nonexcited spins enter the detection slice, which leads to an acceleration of the relaxation time. A model that divides tissue into two compartments is adapted to slice selective inversion in order to derive a simple expression for perfusion-corrected RBV. The aim of the study is to demonstrate the feasibility and accuracy of this technique for quantification of RBV in rat myocardium in vivo. RBV maps were obtained for five rats, and the reproducibility was determined by repeating the experiment several times. A mean RBV value of 12.8 +/- 0.7% (v/v) over all animals was obtained in the myocardium. The results were compared with RBV maps obtained with perfusion-sensitive RBV imaging in the same five rats and with first-pass RBV studies. In order to demonstrate the strength of the new method the vasodilator adenosine was administered and alterations in microcirculation were imaged. Magn Reson Med 42:500-506, 1999.

Adenosine↗

Assessing microsatellite instability with semiautomated fluorescent technology: application to the analysis of primary brain tumors.

The replication error phenotype, revealed by the observation of widespread microsatellite instability (MIN), has been identified as a new mechanism of cancer susceptibility, and the comparison of the allele sizes of polymorphic microsatellite repeats between normal and tumor DNA is now frequently undertaken in colorectal and other human neoplasias. The lack of precise characterization of the electrophoretic profiles of microsatellites is one of the main sources of discord between the rate of MIN reported for the same type of tumor by different investigators. The recent introduction of fluorescent-based semiautomated microsatellite analysis allows a more accurate size comparison, but one or more artificial peaks, generated during polymerase chain reaction (PCR) and/or electrophoresis, are frequently detected along with the true allele peaks. The aim of this study was to characterize the most frequent artificial extra peaks in the short tandem repeats (STRs) used by us to assess MIN in human cancers. We analyzed eight microsatellite loci in 113 primary brain tumors. HumFibra/FGA exhibited the most frequent extra peak formation. For each microsatellite there is a characteristic pattern of artifact formation which must be recognized to avoid a false-positive diagnosis of MIN.

Automation↗

Assessment of thermal tissue ablation with MR elastography.

An important part of thermal ablation therapy is the assessment of the spatial extent of tissue coagulation. In this work, the mechanical properties of thermally-ablated tissue were quantitatively evaluated using magnetic resonance elastography (MRE). This study shows that the mechanical properties of focused ultrasound ablated tissue are significantly different from normal tissue, and the difference can be imaged and measured using MRE. Repeated experiments revealed a reproducible pattern of tissue mechanical property change during thermal ablation in ex vivo bovine muscle. This pattern may reflect changes in intrinsic tissue structure and could be used to evaluate tissue coagulation during thermal ablation therapy. Magn Reson Med 45:80-87, 2001.

Animals↗

Fabrication and characterization of poly(methyl methacrylate) microchannels by in situ polymerization with a novel metal template.

A stainless steel template for the fabrication of plastic microfluidic devices has been developed by photolithography and chemical etching technique. The preparation process of the template is simple, rapid, and low-cost. The cross sectional profiles of raised microchannels on the template are trapezoidal. The surface roughness of the templates was controlled down to 190 nm. The template can be used repeatedly to generate devices reproducibly. The microfluidic devices of poly(methyl methacrylate) (PMMA) were fabricated by in situ polymerization using the templates. The reproducibility of the fabricated microchannel is high and the relative standard deviation is 0.7% by the in situ polymerization approach. Some physical properties of the polymerized microchannels were characterized including the transparency, the thermal deformation temperature, and the dimensional information. Current monitoring was used to evaluate the electroosmotic flow within the microchannels under the electric field strength of 300 V/cm.

Equipment Design↗

On-line pervaporation-capillary electrophoresis for the determination of volatile acidity and free sulfur dioxide in wines.

Pervaporation has been coupled on-line to capillary electrophoresis (CE) by a simple interface consisting of a modified CE vial. The approach allows volatile analytes to be removed and injected into the capillary meanwhile the sample matrix remains in the pervaporator. By this approach volatile acidity and free sulfur dioxide have been simultaneously determined in wines. The detection limits (LODs) are 1.25 and 5.00 microg/mL, the quantification limits 4.12 and 16.50 microg/mL, and the linear dynamic ranges between LOD and 50 microg/mL and between 0.1 and 0.9 g/L for free sulfur dioxide and volatile acidity, respectively. The repeatability and within laboratory reproducibility, expressed as relative standard deviation (RSD), are 1.61% and 3.00% for free sulfur, and 3.35% and 4.58% for volatile acidity, respectively. The optimal pervaporation time and the time necessary for the individual separation-detection of the target analytes are 6 and 5 min, respectively. The analysis frequency is 7 h(-1) and the sample amount necessary is less than 7 mL. The proposed method and official methods for the analytes were applied to 32 wine samples. A two-tailed t-test was used to compare the methods, which yielded similar results. The errors, expressed as RSD for the two parameters, ranged between 1.3 and 4.1%.

Acids↗

Effect of estradiol upon serum enzymes in primary biliary cirrhosis.

The repeated observation of a fall in serum enzymes at midmenstrual cycle in an untreated patient with primary biliary cirrhosis stimulated a study of estrogen administration in five patients with primary biliary cirrhosis. One patient was premenopausal, one patient was postmenopausal and three had had oophorectomy. After 2 weeks of ethinyl estradiol, AST was under 100 IU per dl in all and had decreased by 50% or more in 4 of 5 patients. gamma-glutamyltransferase and alkaline phosphatase fell by 50 and 30% or more, respectively, in all patients. The decreases in serum enzymes were statistically significant in all patients for gamma-glutamyltransferase, in 4 of 5 for AST and in 3 of 5 for alkaline phosphatase. One patient developed increased icterus leading to withdrawal of estradiol. Withdrawal of estradiol was followed by return toward control values over variable periods, usually 1 to 4 weeks. Repeated courses of estradiol reproduced these enzyme changes. These observations indicate that estradiol in the doses used (0.05 mg per day) reversibly lowers serum enzyme values in biliary cirrhosis. The mechanism of the effect is unexplained, but an immune system alteration may be responsible.

Adult↗

Validation for tremor quantification of an electromagnetic tracking device.

An electromagnetic tracking system was used to record arm motion in subjects with Parkinson's disease (n = 23), essential tremor (n = 28) or without neurological disease (n = 4). Tremor magnitude was calculated by averaging the three-dimensional displacement of individual tremor bursts. Tremor magnitude calculated in this manner was quite closely correlated with a clinician's estimate (r = 0.88 and 0.86 for Parkinsonian and essential tremors, respectively) and was reproducible (r = 0.93 for repeated recordings). The accuracy of the device and algorithm was confirmed by mechanically generating oscillations of known magnitudes and frequencies. This device is adaptable for quantifying different types of tremors and its accuracy is easy to verify. Because position rather than acceleration is tracked, tremor amplitude can be stated in readily comprehensible units.

Adult↗

Number and relative size of thenar motor units estimated by an adapted multiple point stimulation method.

An adapted multiple point stimulation (AMPS) method is described for estimating the number and relative size of thenar motor units. With this method, the median nerve was stimulated at various sites from the wrist to the elbow. To avoid alternation, only two or three clearly identifiable surface-recorded motor unit action potentials (S-MUAPs) were recruited at each point by incremental stimulation. A total of 10 S-MUAPs, elicited from four to five distinct stimulation points, was used to calculate the average S-MUAP size. By dividing the maximum M-potential size by that value, a motor unit number estimate (MUNE) was derived. In 59 healthy volunteers, from 19 to 87 years old, the mean average S-MUAP size was 87 +/- 27.6 microV.ms and the mean MUNE was 278 +/- 113 motor units. When performed repeatedly, the results were reproducible. The number of motor units declined exponentially with age while average S-MUAP sizes increased only moderately. To assess the validity of the AMPS method, its results were correlated with those obtained using the F-response technique. The correlation coefficient was 0.83 (P < 0.001).

Action Potentials↗

Optimisation of the quantitative determination of chlormequat by matrix-assisted laser desorption/ionisation mass spectrometry.

The plant growth regulator chlormequat, an involatile quaternary ammonium salt, has been quantified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). Restrictions for quantitative MALDI-TOFMS analysis, such as irreproducible crystallisation and unsatisfactory laser stability, have been overcome by the application of two synthesised isotopically labelled standards and the optimisation of the measurement protocol. Data acquisition at constant laser power was compared to data acquisition at approximately constant ion abundance of the relevant ions (analyte and internal standards). Data acquisition at constant ion abundance performed better and enabled a high number of consecutive firings to the same sample deposition area. Furthermore an increased sample-to-sample repeatability and a high reproducibility over several weeks without re-calibration have been attained by this method. Linearity over three orders of magnitude (0.05 to 30 ng/microL chlormequat), with a correlation coefficient of 0.9997, was achieved using [13C3]-chlormequat as internal standard. Limit of detection and limit of determination were determined to be in the low pg/microL range for pure standard solutions. Thin-layer chromatography was applied for the removal of high amounts of choline, which is often present in plant tissue extracts and can adversely affect the ionisation and detection of chlormequat by MALDI-TOFMS. The use of two internal standards ([13C3]- and [2H9]-chlormequat) enabled direct quantification and simultaneous control of the recovery.

Calibration↗

Kr-81m for determination of right ventricular ejection fraction (RVEF).

A 10--12 mCi 81Rb leads to 81mKr generator was connected to a specially designed short-period infusion set, to produce an equilibrium activity distribution in the right heart. This procedure was tested in 25 individuals to calculate the right ventricular ejection fraction (RVEF). On average 30 heart cycles were analyzed per study. No background activity from the left heart was visualized because of the radionuclide exhalation. The background from the lungs could be neglected, which is partially due to the ultra-short half-life of the nuclide (t 1/2 = 13 S). Thus, an easy automatic procedure can be applied to delineate the ventricle and to calculate the RVEF. The data showed excellent reproducibility, when investigations were repeated. The method would benefit from use of higher activity generators.

Adult↗

Complex-irreversibility and evolution.

Both, irreversibility and evolution, imply order in time. It is argued that the only possible concept of time is a 'system-specific time', and that order in time is convertible into order in space and vice-versa. While life-less, complex systems are irreversible because of their complexity and, hence, not repeatable, living systems are reproduced by irreversible copy-reproduction and by coding. This mode of reproduction results of necessity in an arrow of time of growth and increasing complexity with death as its antagonist, and in obligatory spatial asymmetry. This arrow of increasing organic complexity is simultaneous with, and independent of, the arrow of increasing entropy. - A generalized, organic hierarchy is proposed as the model to study higher evolution. This hierarchy reproduces itself by differential rates of reproduction of its subunits within and between the various hierarchical levels of organization. Phylogenetic change is brought about by a change in this hierarchy's specific phase pattern of growth. Continuous and discrete organization is defined, and it is shown that specific relations between continuous and discrete levels within the hierarchy result in accumulation of neutral alleles. This accumulation is due to complex-irreversibility and causes genetic stabilisation, i.e. heritability, of the species-specific morphology of organisms.

Animals↗

Oxygen consumption and metabolic strain in rowing ergometer exercise.

Oxygen consumption (VO2) when rowing was determined on a mechanically braked rowing ergometer (RE) with an electronic measuring device. VO2 was measured by an open spirometric system. The pneumotachograph valve was fixed to the sliding seat, thus reducing movement artefacts. A multi-stage test was performed, beginning with a work load of 150 W and increasing by 50 W every 2 minutes up to exhaustion. Serum lactate concentrations were determined in a 30 s break between the work stages. 61 examinations of oarsmen performing at maximum power of 5 W X kg-1 or more were analysed VO2 and heart rate (HR) for each working stage were measured and the regression line of VO2 on the work load (P) and an estimation error (Sxy) were calculated: VO2 = 12.5 X P + 415.2 (ml X min-1) (Sxy = +/- 337 ml, r = 0.98) Good reproducibility was found in repeated examinations. Similar spiroergometry was carried out on a bicycle ergometer (BE) with 10 well trained rowers and 6 trained cyclists. VO2 of rowing was about 600 ml X min-1 higher than for bicycling in the submaximal stages for both groups. The VO2max of RE exercise was 2.6% higher than for oarsmen on BE, and the cyclists reached a greater VO2 on BE than the oarsmen. No differences were found between RE and BE exercise heart rate. The net work efficiency when rowing was 19% for both groups, experienced and inexperienced: when cycling it was 25% for cyclists and 23% for oarsmen.

Energy Metabolism↗

Facilitation of ethanol consumption by intracerebroventricular infusions of corticosterone.

Male Wistar rats bearing intracerebroventricular (ICV) cannulae and with simultaneous access to 6% ethanol and water were subjected to adrenalectomy (ADX) or sham surgery. ADX decreased ethanol intake. Starting a few days later, the animals received ICV infusions with 100 micrograms corticosterone acetate (CORT) with 2-to 3-day intervals for 2 weeks. ICV CORT, but not SC CORT at the same dose, restored ethanol consumption in ADX rats to preoperative levels, whereas vehicle infusions (propylene glycol) did not. Adrenally intact animals, which normally consumed moderate amounts of ethanol (approximately 0.5 g/kg per day), also showed a robust effect of ICV infusions of CORT, whereas this facilitatory effect was not observed in high consumers (approximately 3.0 g/kg per day). The suppressive effect of ADX on ethanol intake was not reproduced by concurrent and repeated ICV infusions of intracellular mineralocorticoid (RU 28318) and glucocorticoid (mifepristone) receptor blockers. It is concluded that CORT stimulates alcohol consumption by acting in the brain, probably by way of neuronal membrane mechanisms.

Adrenalectomy↗

Identification of victims of the 1998 Taoyuan Airbus crash accident using DNA analysis.

In February 1998 a civilian aeroplane carrying 196 individuals crashed in Taiwan and killed another 6 people on the ground. Although there were dental and medical records, fingerprints, photographic evidence and personal effects to identify some of the victims, DNA analysis was required to further identify severely damaged remains. From the 202 people known to have perished in the plane crash, a total of 685 fragments of human remains were subjected to DNA analysis. The analysis was carried out using nine microsatellite loci, plus amelogenin to cluster the 685 fragments into 202 groups, accounting for all the victims. To establish genetic relatedness of the victims to other victims and living relatives, additional DNA loci were used. In this case the paternity index was increased by using HLA DQA1 plus Polymarker. The same 16 DNA loci were used to test blood samples from 201 relatives to establish parent/child and sibling relationships. With the exception of 19 victims identified by non-genetic evidence, 183 victims were successfully identified by DNA typing with relatively high values of paternity index by the direct or indirect comparison of relatives. The 202 victims were from 37 different families, ranging in size from 2 to 13 members and 74 individuals known to be unrelated to any other victim. The DNA from living relatives was used to identify one member of a family group, from which other victims of the family could be identified. ABO blood group information was further used to confirm genetic relatedness within families. A comparison of the DNA profiling results to the ABO blood group of the victims showed no discrepancies with the exception of two mutations in the FGA locus. In cases of severely damaged victims from a plane crash, DNA analysis proved to be the best choice to identify victims.

Accidents, Aviation↗