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Miniaturization of batch- and flow-type chemiluminescence detectors in capillary electrophoresis.

Glass and PTFE tubes as detection cells were put in small light-tight boxes to achieve miniaturization of batch-and flow-type chemiluminescence detectors for capillary electrophoresis. These light-tight boxes which included a detection cell and a photosensor module were successfully designed. In the batch-type detector using a glass tube as a detection cell, the influences of a repeated injection of sample and a reagent volume of the detection cell on chemiluminescence intensity were examined in detail. By using 3.8 mm I.D. glass tube including 400 microl chemiluminescence reagent solution, the chemiluminescence peaks were reproducibly observed for the repeated injection experiment up to the eight injection with each run time of 3.0 min. Dansyl-Trp was determined over the range 3 x 10(-8)-1 x 10(-5) M with the detection limit of 0.43 fmol (S/N=3). In the flow-type detector using a PTFE tube as a detection cell, both ends of the PTFE tube were connected to three-way joints; a chemiluminescence reagent solution was delivered into the cell and a capillary was inserted through one of the joints while an electrode was inserted through the other one. Dansyl-Trp was determined over the range 1 x 10(-7)-1 x 10(-5) M with the detection limit of 1.3 fmol (S/N=3). By using the compact flow-type detector, a mixture of dansyl-amino acids was separated and detected in micellar electrokinetic chromatography mode.

Amino Acids↗

[Development of a questionnaire concerning the knowledge of COPD].

INTRODUCTION: The level of a patient's knowledge about his disease and its treatment is an essential part of an educational assessment. It is useful therefore to make use of a rapid, easy and valid method to collect the information necessary to develop an educational programme adapted to the needs of the patient. The aim of this study is to validate, in a structured way, a knowledge questionnaire on chronic obstructive pulmonary disease (COPD). METHODS: Following a revue of the literature an initial questionnaire was constructed. It included of four domaines: biomedical aspects; symptoms and signs of severity; general knowledge and treatments. The questionnaire was tested on 35 subjects with COPD in order to assess its clarity and comprehensibility. It was reviewed and modified in both content and format by 11: French experts. The reproducibility was studied by repeat testing. RESULTS: The first version of the questionnaire developed by the working party consisted of 50 items. It was reduced to 41 items after interviews with 35 COPD patients and evaluation by 11 experts. The questionnaire appeared to be reproducible: mean concordance 79.5%; minimum 53.3%; maximum 100% and intra-class correlation coefficient 0.53. CONCLUSION: This study lead to the development of a French language COPD knowledge questionnaire.

Adult↗

Collaborative study for the establishment of the Ph. Eur. BRP for oral poliomyelitis vaccine (OPV) Batch 3 for use in the potency assay.

A collaborative study was initiated by the European Directorate for the Quality of Medicines (EDQM) with the goal to calibrate the trivalent candidate European Pharmacopoeia Biological Reference Preparation (BRP) for oral poliomyelitis vaccine (OPV) Batch 3 against the 1st International Standard (IS) for OPV and to establish the material as a working standard. The material is a commercial trivalent stabilised oral poliomyelitis vaccine, consisting of Sabin strains of live attenuated poliovirus types 1, 2 and 3. The new standard is meant to replace the current European Pharmacopoeia BRP for OPV Batch 2, the stocks of which will soon be depleted. Fourteen laboratories participated in the study. Three samples had to be assayed (1st IS, BRP Batch 2, candidate BRP Batch 3). The potency of each virus type in each preparation had to be estimated by using either common monoclonal anti-polio antibody sera and/or the participant's routinely used antisera for neutralizing two of the three virus types present in the trivalent vaccine. In addition the total virus content had to be determined. From the raw data returned, log10 CCID50/ml values were calculated using the probit method (CCID50 is the dose infecting 50% of the cell cultures). The precision (intra-assay variation), repeatability (intralaboratory variation) and reproducibility (inter-laboratory variation) were assessed as absolute titres and as the adjusted titres (potencies of the test samples calculated relative to the 1st IS). An analysis of variance was performed to determine if there were significant differences between assays using mono- or polyclonal antisera. The precision, determined as the width of the uncorrected confidence limits taken across all assays within laboratories, varied on average from +/- 0.13 to +/- 0.34. The repeatability (uncorrected titres) standard deviation varied from 0.08 to 0.27 and was on average 0.174. The repeatability (corrected titres) standard deviation varied from 0.11 to 0.33 and was on average 0.176. In both cases the repeatability standard deviation was very similar when compared between types, between antisera and between samples. None of these differences were statistically significant. The reproducibility (for the corrected titres) was analysed by calculating the standard deviation of the laboratory means which ranged from 0.10 to 0.21 and was on average 0.151. For the corrected titres there is no significant indication that the antiserum type used for neutralisation affects the potency estimate. The study shows that the candidate BRP Batch 3 is suitable as a reference substance and, based on the current results, 6.99, 6.06, 6.83 and 7.20 log10 CCID50/ml are the potencies assigned for Types 1, 2, 3 and total virus content, respectively. Stability data indicate that the candidate material is stable when stored at -20 degrees C. Nonetheless the stability of the new reference preparation will be closely monitored. The candidate material was adopted by the European Pharmacopoeia Commission at its session in March 2002 as European Pharmacopoeia OPV BRP Batch 3.

Australia↗

Influence of cytotoxicity and compound precipitation on test results in the alkaline comet assay.

We use the comet assay as part of our genotoxicity screening battery for newly synthesized drug candidates. A dataset of more than 250 tests carried out with 75 drug candidates of various chemical classes was analyzed to elucidate the influence of cytotoxicity and compound precipitation on DNA migration in the comet assay. Using a V79 Chinese hamster cell line, 38 of the compounds were negative and 37 were positive in the comet assay. The reproducibility of test results between repeat experiments was 85%. Data on 72 tests with a negative call in which the compounds were tested up to highly cytotoxic concentrations demonstrated that cytotoxicity, as determined by Trypan blue dye exclusion and occurrence of cells with completely fragmented chromatin, did not lead to false positive test results. The majority (64.2%) of compounds with a positive call induced elevated DNA migration in the absence of excessive cytotoxicity. Compound precipitation was observed in 84 tests. In 88.1% of these cases, the test result at the precipitating concentration did not differ from that found at the highest soluble concentration. Half of the remaining 11.9% of contrary results (most of them weak effects) were not reproducible in the respective repeat experiment, indicating no or only a negligible influence of precipitation on test results. The data indicate that using V79 cells, the comet assay specifically detects genotoxic effects and is not confounded by cytotoxicity or compound precipitation under the conditions used.

Animals↗

Validity and reproducibility of a semi-quantitative food-frequency questionnaire for estimating calcium intake in Belgian preschool children.

Relative validity and reproducibility of a semi-quantitative food-frequency questionnaire (FFQ) for measuring preschool children's usual Ca intake were assessed using parents or guardians as a proxy. Estimated diet records (EDR; 3 d) were used as the reference method and reproducibility was measured by repeated FFQ administrations 5 weeks apart. From 2095 preschool children (2.5-6.5 years) randomly selected in Flanders (Flemish region of Belgium), 1052 returned a FFQ and EDR. Stringent exclusion procedures reduced the sample for validity analyses to 509 children. From a separate sample of 244 preschool children, 124 returned two FFQ, of whom sixty were included in the reproducibility analysis. Mean Ca intakes were 838 (sd 305) and 777 (sd 296) mg/d for EDR and FFQ respectively, indicating a mean difference of 60.9 (sd 294.4) mg/d (P<0.001). Pearson's correlation was 0.52. Cross-classification analysis of the FFQ and EDR classified 83 % of the subjects in the same or adjacent category and 2.4 % in extreme quartiles. Actual values for surrogate FFQ quartiles showed a progressive increase in Ca intake (P<0.001). The FFQ correctly identified 77 % of the children consuming less Ca than the age-specific RDA (800 mg/d). Correlation between repeated administrations was 0.79. No significant difference between mean Ca intakes was established by two administrations (23.8 (sd 161.2) mg/d). Cross-classification of repeated administrations classified 93.4 % of the subjects in the same or adjacent category and no subjects in extreme categories. This FFQ tended to underestimate preschool children's Ca intake when administered by a proxy. However, it demonstrated good repeatability and fairly good ability to classify subjects into extremes of Ca intake.

Belgium↗

D and 18O enrichment measurements in biological fluids in a continuous-flow elemental analyser with an isotope-ratio mass spectrometer using two configurations.

In doubly labelled water studies, biological sample enrichments are mainly measured using off-line techniques (equilibration followed by dual-inlet introduction) or high-temperature elemental analysis (HT-EA), coupled with an isotope-ratio mass spectrometer (IRMS). Here another continuous-flow method, (CF-EA/IRMS), initially dedicated to water, is tested for plasma and urine analyses. The elemental analyser configuration is adapted for each stable isotope: chromium tube for deuterium reduction and glassy carbon reactor for 18O pyrolysis. Before on-line conversion of water into gas, each matrix is submitted to a short and easy treatment, which is the same for the analysis of the two isotopes. Plasma is passed through centrifugal filters. Urine is cleaned with black carbon and filtered (0.45 microm diameter). Tested between 150 and 300 ppm in these fluids, the D/H ratio response is linear with good repeatability (SD<0.2 ppm) and reproducibility (SD<0.5 ppm). For 18O/16O ratios (from 2000 to 2200 ppm), the same repeatability is obtained with a between-day precision lower than 1.4 ppm. The accuracy on biological samples is validated by comparison to classical dual-inlet methods: 18O analyses give more accurate results. The data show that enriched physiological fluids can be successfully analysed in CF-EA/IRMS.

Animals↗

The maximal sniff in the assessment of diaphragm function in man.

We studied diaphragm function in a total of 64 normal subjects, who had no past or present respiratory or neuromuscular impairment. We measured transdiaphragmatic pressure (Pdi) during maximal sniffs and compared these values with Pdi during maximal static inspiratory efforts (PImax.). The range of Pdi during maximal sniffs (82-204 cm H2O) had better defined lower limits than Pdi during PImax. (16-164 cm H2O) and a higher mean value: mean +/- SD for maximal sniffs was 137 +/- 28 cm H2O and for PImax. was 90 +/- 37 cm H2O. The reproducibility of sniff Pdi was assessed in eight randomly chosen subjects over 3 days: the mean coefficient of variation was 7.2%. By comparison the coefficient of variation of Pdi during PImax. was 13.0% in seven subjects. The maximal sniff is a spontaneous manoeuvre, easily performed, repeatable without tiring, and reproducible. Its measurement provides a more reliable quantitative method for assessment of diaphragm strength, which has potential in clinical practice.

Adult↗

Laser scanning as a tool for assessment of HIV-related facial lipoatrophy: evaluation of accuracy and reproducibility.

BACKGROUND: Facial lipoatrophy (LA) is a common complication of highly active antiretroviral therapy (HAART). Research into causes and treatment of facial LA is hindered by the lack of an objective measurement tool. OBJECTIVE: To evaluate the accuracy and reproducibility of three-dimensional laser scanning (LS) for estimating cheek volume changes. METHODS: Paired laser scans were performed and the images superimposed using commercial software. The volume difference between images was computed within a circle of radius 25 mm placed in a standardized position over the cheek area. Accuracy was tested by scanning before and after known volumes of plasticine (0.5--5 mL) were applied to the cheek area of a mannequin to simulate volume change. Reproducibility was tested by repeated scanning of the mannequin with and without 2 mL of plasticine, and repeated scanning of 10 healthy subjects over the course of 1 week. RESULTS: The mean difference between actual and estimated volume change was small across the range of volumes tested [mean difference 0.08 mL; 95% confidence interval (CI)-0.36 to 0.20 mL). The coefficient of variation for repeated measurements of 2-mL volume change was 5.8%. The intraclass correlation coefficient for scan-to-scan variability was 0.812 (95% CI 0.515--0.947) and for day-to-day variability it was 0.764 (95% CI 0.332--0.935). Conclusions LS is an accurate and reproducible method for estimating cheek volume changes. It may be useful as an objective tool for assessment of facial LA in clinical research studies.

Antiretroviral Therapy, Highly Active↗

Gradient echo MRI characterization of development of atherosclerosis in the abdominal aorta in Watanabe Heritable Hyperlipidemic rabbits.

PURPOSE: The Watanabe Heritable Hyperlipidemic (WHHL) rabbit provides an important model of spontaneous atherosclerosis. With a strain of WHHL rabbits which do not develop abdominal aorta lumen stenosis even with advanced atherosclerosis, we studied the MRI-histology correlation, and the natural progression of atherosclerosis in the abdominal aorta. In addition, intra-reader segmentation repeatability and scan-rescan reproducibility were assessed. METHODS: Two batches of female WHHL rabbits were used. The first batch of 6 rabbits was scanned at 20 weeks old. A second batch of 17 rabbits was scanned at 50 weeks old and then randomly divided into two subgroups: 8 were killed for histologic investigation; 9 were kept alive for follow-up, with repeat scanning a week later to assess scan-rescan reproducibility, and again at 73 weeks old to assess disease progression. MR images were acquired at 4.7 T using a chemical shift selective fat suppression gradient echo with a saturation band suppressing blood signal within the aortic lumen. Five slices per animal were acquired, centered around the renal artery region of the abdominal aorta, with in-plane resolution of 0.195 mm and slice thickness of 3 mm. RESULTS: The coefficient of variation for intra-reader reproducibility for aortic wall thickness measurements was 2.5% for repeat segmentations of the same scans on the same day, but segmentations of these same scans made 8 months later showed a systematic change, suggesting that intra-reader bias as well as increased variability could compromise assessments made over time. Comparative analyses were therefore performed in one postprocessing session. The coefficient of variation for scan-rescan reproducibility for aortic wall thickness was 5.5% for nine pairs of scans acquired a week apart and segmented on the same day. Good MRI-histology correlation was obtained. The MRI-measured mean aortic wall thickness of animals at 20 weeks of age was 76% that of animals at 50 weeks of age (p < 0.001). There was a small increase in aortic wall thickness between 50 and 73 weeks of age, but this was not significant (p > 0.05). The corresponding differences in lumen cross-sectional areas at 20, 50, and 73 weeks of age were not significant. These results were consistent with in-house historical histology data on this strain of rabbits. CONCLUSIONS: High-resolution gradient echo MRI can follow disease progression in the WHHL rabbit spontaneous atherosclerosis disease model.

Aging↗

Reproducibility in monitoring nocturnal penile tumescence and rigidity.

The evaluation of sexual dysfunction has improved with the advent of methods to test nocturnal penile tumescence that also monitor penile rigidity. Earlier techniques may not have recorded abnormal rigidity despite normal tumescence. To test the reproducibility of penile tumescence and rigidity, the results of initial and repeat tracings performed a mean of 39 days apart were compared in 17 patients (median age 62 years). Three nocturnal patterns were identified. 1) Among the 17 patients the initial penile tumescence and rigidity pattern was reproduced on repeat testing in 15. 2) Patterns that were not reproduced in the other 2 patients were explained by the ingestion of alcohol or because of a febrile illness during the period monitored. 3) Nocturnal penile rigidity and tumescence tracings from these patients reproduced previous patterns. Monitoring of nocturnal penile tumescence and rigidity is a useful and reproducible tool in the evaluation of male sexual dysfunction.

Adult↗

Regulation of vascular and gastric smooth muscle contractility by pervanadate.

1. The contractile actions of vanadate (VO4) and pervanadate (PV, peroxide(s) of vanadate) were studied in rat gastric longitudinal muscle strips and in aortic rings. The roles of extracellular sodium and calcium were evaluated and the potential effects of nerve-released agonists were considered. The possibility that these responses were due to the potentiation of tyrosine kinase activity, as a result of PV-mediated tyrosine phosphatase inhibition was explored with the use of tyrosine kinase inhibitors (genistein, tyrphostin) and by Western blot analysis of phosphotyrosyl proteins in PV-treated tissues. The ability of PV to mimic the action of the tyrosine kinase receptor-associated agonist, epidermal growth factor-urogastrone (EGF-Uro), in the gastric preparation was also studied. 2. PV caused concentration-dependent contractions in both gastric and aorta-derived tissues, with a potency that was 1 to 2 orders of magnitude greater than that of VO4. 3. Although repeated exposure of gastric and aortic tissues to a fixed concentration of VO4 caused reproducible contractions in both tissues, repeated exposure of gastric tissue to PV caused an increased contractile response plateauing after 3 exposures. In contrast, a single exposure of aortic tissue to PV (20 microM) caused a prolonged desensitization of the tissue to the subsequent contractile actions of PV or other agonists. 4. The contractile responses to PV were unaffected in both preparations by tetrodotoxin, atropine, yohimbine and phenoxybenzamine; and in the aortic preparation, the responses to VO4 and PV were the same in the presence or absence of a functional endothelium. 5. PV-induced contractions in both tissues were observed in the absence of extracellular sodium but required extracellular calcium and were attenuated by 1 micro M nifedipine.6. In the gastric preparation, the characteristics of the contractile actions of PV paralleled those of EGF-Uro in terms of (1) inhibition by genistein, (2) inhibition by indomethacin and (3) a requirement for extracellular calcium. These response characteristics differed from those of other contractile agonists such as carbachol.7. In both the gastric and aortic preparations genistein was able to inhibit PV-induced contractions selectively without causing comparable inhibition of KCI-induced contractions. Tyrphostin (AG18) also selectively blocked PV-induced contractions in the gastric, but not in the aortic preparation.8. In both the gastric and aortic tissue, in step with an increased contractile response, PV caused increases in tissue phosphotyrosyl protein content, as detected by Western blot analysis using a monoclonal antiphosphotyrosine antibody; the increases in phosphotyrosyl protein content were reduced when tissues were treated with PV at the same time as a tyrosine kinase inhibitor.9 PV, at sub-contractile concentrations, potentiated the contractile action of angiotensin II in both the gastric and aorta tissue.10 We conclude that the growth factor-mimetic agent, PV, is a much more potent contractile agonist than V04 in both vascular and gastric smooth muscle tissue. PV can cause enhanced tissue phosphotyrosyl protein content most likely via the inhibition of tissue protein tyrosine phosphatases. The contractile actions of PV, which require extracelullar calcium and are independent of extracellular sodium, would appear not to be due either to Na+/Ca2" exchange, promoted by Na+/K+-ATPase inhibition or to the inhibition of Ca2+-ATPase and might be best explained by the ability of PV, via tyrosine phosphatase inhibition, to potentiate a tyrosine kinase pathway linked to calcium entry and to the contractile process.

Angiotensin II↗

Inflammatory pain in experimental burns in man.

Human experimental pain models are important tools in pain research. The primary aims of pain research in normal man is 1) to provide insight in pain mechanisms, 2) to provide a rational basis for clinical trials of pain relieving interventions, and 3) to confirm the anti-nociceptive effects demonstrated in animal models. Most often clinical pain is due to tissue damage leading to acute inflammation and hyperalgesia, but only few human pain models have examined pain responses in injured tissues. Therefore, models with controlled and reversible tissue trauma are needed. The human burn model is an example of such a model, and several groups have performed studies of analgesics and pain mechanisms based on the model. The thesis aims to provide a critical review of the human burn model as a tool in pain research, and to give suggestions for development of the model and future research. The pain and inflammatory responses to superficial thermal burns in skin have been studied in healthy volunteers. Burns have the potential for releasing most of the inflammatory and chemical mediators that produce sensitisation and excitation of nociceptors, and the intense nociceptive input during injury produces sensitisation of central neurones in the nociceptive pathway. Pain and hyperalgesia have been evaluated in the model by thermal, various mechanical, and electrical stimuli. The different methods of pain assessments are discussed to clarify the underlying neural mechanisms, the questions that can be addressed by the measurements, and the discrepancies in results between studies. Inflammation has been evaluated in the model by skin erythema intensity, area of flare, and blister formation. The major determinant of skin erythema intensity is the amount of blood in the most superficial part of the dermis, and burn-induced erythema may be primarily due to congestion of capillary loops and postcapillary venules. The area of flare may be used to evaluate the efferent function of heat-sensitive A delta- and C-fibre nociceptors, whereas blisters may be used to assess edema formation and the degree of injury. Hyperalgesia is induced immediately by the burns and lasts about 24 h dependent on the intensity of the heat stimulus. The burns heal without sequela. A study of the reproducibility of pain assessments in the burn model has shown that measures based on repeated measurements were significantly more reproducible than measures based on single time points. Further, within-day reproducibility was better than between-day reproducibility. Within-day variations of heat pain responses to 45 degrees C and 47 degrees C were smaller than that of pain responses to 43 degrees C, suggesting that assessments using clearly painful stimuli may be more reproducible. A methodological study also demonstrated that habituation to experimental pain developed as the study proceeded. Habituation is common in experimental pain models, and dividing analgesics and placebo evenly between the study days is one way of eliminating the effects of habituation. The use of simultaneous right-left comparisons represents the ideal design when possible. The burn model has been a valuable tool in the study of pain mechanisms. Hyperalgesia to heat in the burned area (primary hyperalgesia) is mediated by sensitisation of C-fibre mechano-heat-sensitive (CMH) nociceptors and A delta-fibre mechano-heat-sensitive (AMH) nociceptors of type I in hairy skin. A contribution from sensitised CNS neurones is likely, and the sensitisation of nociceptors is confined to the injured area. The presence of hyperalgesia to heat in normal skin surrounding a burn (secondary hyperalgesia) has been demonstrated in several studies, but the pain threshold may be unaltered. The mechanisms for primary hyperalgesia to mechanical stimuli may be both peripheral and central, but the importance of peripheral mechanisms is unclear and central mechanisms may account for mechanical hyperalgesia in both the primary and th

Algorithms↗

Accuracy and repeatability of Roentgen stereophotogrammetric analysis (RSA) for measuring knee laxity in longitudinal studies.

Roentgen stereophotogrammetric analysis (RSA) can be used to assess temporal changes in anterior-posterior (A-P) knee laxity. However, the accuracy and precision of RSA is dependent on many factors and should be independently evaluated for a particular application. The objective of this study was to evaluate the use of RSA for measuring A-P knee laxity. The specific aims were to assess the variation or "noise" inherent to RSA, to determine the reproducibility of RSA for repeated A-P laxity testing, and to assess the accuracy of these measurements. Two experiments were performed. The first experiment utilized three rigid models of the tibiofemoral joint to assess the noise and to compare digitization errors of two independent examiners. No differences were found in the kinematic outputs of the RSA due to examiner, repeated trials, or the model used. In a second experiment, A-P laxity values between the A-P shear load limits of +/-60 N of five cadaver goat knees were measured to assess the error associated with repeated testing. The RSA laxity values were also compared to those obtained from a custom designed linkage system. The mean A-P laxity values with the knee 30 degrees, 60 degrees, and 90 degrees of flexion for the ACL-intact goat knee (+/-95% confidence interval) were 0.8 (+/-0.25), 0.9 (+/-0.29), and 0.4 (+/-0.22) mm, respectively. In the ACL-deficient knee, the A-P laxity values increased by an order of magnitude to 8.8 (+/-1.39), 7.6 (+/-1.32), and 3.1 (+/-1.20)mm, respectively. No significant differences were found between the A-P laxity values measured by RSA and the independent measurement technique. A highly significant linear relationship (r(2)=0.83) was also found between these techniques. This study suggests that the RSA method is an accurate and precise means to measure A-P knee laxity for repeated testing over time.

Animals↗

[Evaluation of the effectiveness of corinfar in patients with ischemic heart disease using a method of repeated transesophageal electric stimulation of the left atrium].

The efficiency of 20 mg corinfar, administered sublingually, was assessed by means of esophageal left-atrial electrostimulation (EES) in 49 coronary patients with angina of functional classes 1 through 4. The antiischemic effect of corinfar was evaluated within 30 min after the administration in 27 patients and within 1 hour after the administration in 22 patients. To assess the reproducibility of the results, repeated EES was performed in 11 patients, treated with placebo this time. Corinfar was shown to have a good antiischemic effect 1 hour after sublingual administration, with the maximum induced heart rate rising from 130 to 147 beats per minute and the total duration of the stimulation increasing from 232 to 308 seconds. It is demonstrated that paired EES may be used for the assessment of corinfar efficiency in coronary patients.

Adult↗

Significance of the walk-through angina phenomenon during exercise testing.

Out of 3,900 patients who performed an exercise test at our clinic, 3 patients demonstrated a walk-through phenomenon (WTP), defined as the occurrence of mild angina during the first stages of exercise with disappearance of chest pain at higher workloads despite a greater pressure-rate product. 2 patients had variant angina, one with normal coronary arteries and the other with single vessel disease, while the third patient had stable exertional angina and a severe coronary artery disease with occlusion of two major vessels retrogradely filled by collateral channels. Repeat exercise tests failed to reproduce constantly the WTP in the 2 patients with variant angina, while in the third patient the phenomenon was repeatedly induced by exercise testing. Thus the WTP, although rarely found during exercise testing, can be observed in two subsets of patients. In variant angina the WTP is not reproducible and is probably due to coronary spasm, spontaneously subsiding during exercise. In patients with exertional angina and severe coronary artery disease, the WTP can be repeatedly observed during exercise and is likely to be secondary to a delayed vasodilation of collateral vessels. The clinical characteristics of the patients and the response to repeat exercise tests may be useful in identifying the different pathogenetic mechanisms.

Adult↗

Comparison of immediate versus day to day variability of ventricular tachycardia induction by programmed stimulation.

The use of programmed stimulation to assess long-term oral antiarrhythmic drug efficacy for ventricular tachycardia is complicated by the fact that mode of ventricular tachycardia induction varies from day to day in the absence of drug therapy. The purpose of this prospective study was to assess whether mode of ventricular tachycardia induction is more reproducible within one study than from day to day. Thirty-nine consecutive patients with documented sustained ventricular tachyarrhythmias secondary to coronary artery disease underwent three inductions of ventricular tachycardia at 15 min intervals in the absence of drug therapy. A stimulation protocol in which the only major variable was the number of extrastimuli required for tachycardia induction was used. Subsequent day to day variability in mode of tachycardia induction was also assessed in the same patients at two further drug-free inductions at intervals of 5 +/- 2 days. The number of extrastimuli required for tachycardia induction was significantly more reproducible at the immediate repeat studies than from day to day (69% of patients versus 31%, p less than 0.01). From these data, probability tables were derived that show the likelihood that changes in inducibility at subsequent tachycardia inductions are due to chance. The QRS configuration of induced ventricular tachycardia was also more reproducible at the immediate studies (64% versus 26%, p less than 0.01). Basic electrophysiologic and stimulation variables differed over a significantly wider range from day to day than at the immediate studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Reproducibility of skeletal muscle vasodilatation responses to Stroop mental challenge over repeated sessions.

Skeletal muscle blood flow responses to stress have implications for psychobiological disease pathways. An important assumption underlying psychophysiological studies relating stress reactivity with disease risk is that individuals are characterized by stable response profiles that can be reliably assessed using acute psychophysiological stress testing. We examined the reproducibility of forearm vasodilatation, blood pressure, and cardiac responses to a 2 min Stroop mental challenge over two repeated stress sessions that were on average 3.6 months apart. Participants were 21 healthy men and women (aged 21.8+/-3.7 years). Vasodilatation, blood pressure and heart rate responses displayed no habituation between sessions, although there was significantly greater cardiac parasympathetic involvement during the second testing session. Significant test-retest correlations between the sessions were observed for both forearm blood flow and heart rate reactivity. These findings demonstrate skeletal muscle vasodilatation responses to repeated stress are robust, so may be a useful psychophysiological indicator in studies of stress reactivity and disease risk.

Adolescent↗

Procoagulant activity of PPD-stimulated human lymphocytes after cryopreservation.

OBJECTIVE: Human mononuclear cells from a previously sensitized donor generate procoagulant activity (PCA) following stimulation with purified protein derivative (PPD). Lymphocytes of tuberculous pleural effusions are also highly responsive to PPD stimulation. We examined the influence of cryopreservation on lymphocytes following stimulation with PPD. DESIGN: Peripheral blood lymphocytes of 5 healthy PPD skin test positive subjects were incubated with either PPD, thromboplastin, or concanavalin A (Con A) at concentrations of 0, 1, and 10 micrograms/ml. PCA was determined by measuring the recalcification time. Tests were repeated following cryopreservation for 4 weeks. RESULTS: Incubation of fresh lymphocytes led to a dose dependent shortening of recalcification time: PPD (0-1-10 micrograms/ml: 100-84-65%), thromboplastin (0-1-10 micrograms/ml: 100-85-62%), and Con A (0-1-10 micrograms/ml: 100-85-42%). These results were highly reproducible when tests were repeated 6 weeks later. Cryopreservation did not significantly affect the expression of PCA following incubation with PPD and with thromboplastin. In contrast, cryopreservation significantly diminished the degree of Con A generated PCA. CONCLUSION: Cryopreservation and storage of human lymphocytes is possible without alteration of PCA expression following their incubation with PPD or thromboplastin.

Blood Coagulation Factors↗