Patch testing with thin-layer chromatograms.
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Biomedical subjects
Publications and source records attributed to M Frick.
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OBJECTIVES: To evaluate the duration of anticoagulation treatment with warfarin sodium before elective DC-cardioversion and to identify clinical variables predicting short-term versus long-term waiting times. DESIGN: Retrospective. SUBJECTS: Patients with a known start date for warfarin sodium, a known duration of atrial fibrillation (AF) and who underwent DC-cardioversion were included. MAIN OUTCOME MEASURES: Duration of treatment with warfarin sodium prior to DC-cardioversion. METHODS: The hospital records of 288 consecutive patients with AF scheduled for elective cardioversion at two hospitals in Stockholm were reviewed. Only patients with a known start date for warfarin sodium and known duration of AF were included in the study. RESULTS: The median age was 70 (26-85) years and the duration of AF at time of cardioversion were 18 weeks (5-273) weeks. The median treatment duration prior to cardioversion with warfarin sodium was 12 weeks. Sinus rhythm was established in 224 (78%) patients of which 90 (40%) remained in sinus rhythm 1 month after cardioversion. In multivariate analysis, the only independent predictor of short waiting times for cardioversion (8 vs. 15 weeks) was if a cardiologist instituted the treatment with warfarin sodium (P < 0.001, 95% CI 5.0-9.0). CONCLUSION: The average waiting time from start of warfarin sodium treatment to elective cardioversion exceeds by far the recommended 3-4 weeks on therapeutic international normalized ratio (INR). In order to minimize the time period until cardioversion significant changes in the out-of-hospital care logistics has to be undertaken.
In alveolar type II cells, the release of surfactant is considerably delayed after the formation of exocytotic fusion pores, suggesting that content dispersal may be limited by fusion pore diameter and subject to regulation at a postfusion level. To address this issue, we used confocal FRAP and N-(3-triethylammoniumpropyl)-4-(4-[dibutylamino]styryl) pyridinium dibromide (FM 1-43), a dye yielding intense localized fluorescence of surfactant when entering the vesicle lumen through the fusion pore (Haller, T., J. Ortmayr, F. Friedrich, H. Volkl, and P. Dietl. 1998. Proc. Natl. Acad. Sci. USA. 95:1579-1584). Thus, we have been able to monitor the dynamics of individual fusion pores up to hours in intact cells, and to calculate pore diameters using a diffusion model derived from Fick's law. After formation, fusion pores were arrested in a state impeding the release of vesicle contents, and expanded at irregular times thereafter. The expansion rate of initial pores and the probability of late expansions were increased by elevation of the cytoplasmic Ca2+ concentration. Consistently, content release correlated with the occurrence of Ca2+ oscillations in ATP-treated cells, and expanded fusion pores were detectable by EM. This study supports a new concept in exocytosis, implicating fusion pores in the regulation of content release for extended periods after initial formation.
Purinergic stimulation of surfactant secretion via exocytosis of lamellar bodies is mediated by an elevation of the intracellular Ca2+ concentration ([Ca2+](i)). We tested the dihydropyridine (DHP) analogues isradipine (+/-enantiomers), nifedipine and Bay K 8644 (racemic forms) on ATP-induced surfactant secretion and [Ca2+](i) in single type II cells, using FM1-43 and fura-2 fluorescence. None of the DHPs (2 microM) had an effect on ATP-induced surfactant secretion in the dark. They did, however, inhibit secretion in a concentration-dependent manner during illumination, particularly with UV light. This effect was not stereospecific, because it was mimicked by (-)-isradipine. In addition, (+)- or (-)-isradipine, but not nifedipine or Bay K 8644, elicited a slow increase of [Ca2+](i) during illumination with UV light, which was reversible by exposure to dark. None of the DHPs inhibited the ATP-induced Ca2+ signal. In perforated patch clamp experiments, depolarizing voltage steps did not induce L-type Ca2+ (Sr(2+)) currents, even in the presence of the agonist Bay K 8644 (1 microM). We conclude that impairment of ATP-induced surfactant secretion by all tested DHPs and alterations of Ca2+ homeostasis by isradipine are photoactivated effects, independent of L-type Ca2+ channels.
BACKGROUND: The recurrence rate of atrial fibrillation (AF) after elective cardioversion is high. HYPOTHESIS: The study aimed to identify clinical predictors for successful electrical cardioversion and maintenance of sinus rhythm after a first electrical cardioversion in patients with persistent AF without concomitant antiarrhythmic drugs of class I and III. METHODS: Consecutive outpatients (n = 166) with persistent AF for > 1 month, scheduled for elective cardioversion, were prospectively included in the study. A clinical investigation, echocardiographic assay, and Holter electrocardiogram (ECG) before and ECG 4 weeks after cardioversion, were performed in all patients. RESULTS: The mean age of the patients was 68 years (range 45-83) and duration of AF was 5 (1-48) months. Sinus rhythm was established in 124 (75%) patients. In multivariate analysis, only duration of AF < 6 months (p < 0.04, odds ratio [OR] 2.2, 95% confidence interval [CI] 1.1 to 4.7) and patients weight (p < 0.03, OR 2.3, 95% CI 1.1 to 4.8 for weight < 80 kg) were identified as independent predictors of successful cardioversion. At 4 weeks after cardioversion, only 46 (37%) of 124 patients maintained sinus rhythm. Independent factors for maintenance of sinus rhythm, in multivariate analysis, were AF <3 months (p < 0.04, OR 2.5, 95% CI 1.1 to 5.6), treatment with beta blockers (p < 0.00001, OR 7.0, 95% CI 3.0 to 16.3) or verapamil/diltiazem (p < 0.04, OR 3.6, 95% CI 1.1 to 12.1), and right atrial dimension < 37 mm (p < 0.02, OR 5.9, 95% CI 1.4 to 25.4). CONCLUSIONS: In patients with persistent AF, the patient's weight and the duration of AF are independent predictors for a successful cardioversion. Short duration of AF, treatment with beta blockers or verapamil/diltiazem, and right atrial area/dimension are independent predictors for maintenance of sinus rhythm.
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OBJECTIVE: This prospective study was designed to investigate the differences between asymptomatic versus symptomatic arrhythmia as well as left ventricular dysfunction in a consecutive population of patients with persistent atrial fibrillation. DESIGN: A total of 282 consecutive outpatients referred with persistent atrial fibrillation formed the study population. A structured medical history was obtained. A two-dimensional transthoracic echocardiography to assess the left ventricular function and a 24-h electrocardiogram (ECG) recording were performed. Irregularity of the heart rhythm was analysed with heart rate variability (HRV) in the time domain as well as maximum and minimum heart rate and the longest pause. SETTING: Three university hospitals. RESULTS: The mean age of the patients was 69 years and the mean duration of atrial fibrillation was 7 months. The prevalence of symptomatic patients was 68%, while 32% had no symptoms from atrial fibrillation, left ventricular dysfunction was observed in 20%. Asymptomatic subjects had more often lone atrial fibrillation than those with symptoms. Valvular heart disease was an independent predictor of symptoms while male gender, ischaemic heart disease and a high heart rate were independent predictors of impaired left ventricular function. CONCLUSION: Valvular heart disease is related to symptoms in persistent atrial fibrillation. Ischaemic heart disease, male gender and a high heart rate are more common in patients with impaired left ventricular function. Compromised left ventricular function does, occur also in asymptomatic subjects underlining the importance of a careful investigation including echocardiography in all subjects with persistent atrial fibrillation.
In patients with thrombolyzed acute myocardial infarction, early assessment of the final infarct size is difficult because spontaneous recovery of perfusion and function of the left ventricle may be delayed. This study was undertaken to evaluate the ability of predischarge low-dose dobutamine echocardiography to predict late spontaneous recovery of perfusion assessed by single-photon emission computed tomography after acute myocardial infarction. We prospectively studied 53 consecutive patients with myocardial infarction treated with thrombolysis. Low-dose dobutamine echocardiography and resting (99m)Tc-sestamibi single-photon emission computed tomography (MIBI SPECT) were performed 4 +/- 2 days after infarction. A follow-up SPECT study was carried out in 45 patients after 6 months. Myocardial recovery was defined as a reduction of SPECT defect size by more than 10% at follow-up compared with the early study. In 25 of the 45 patients, the size of the left ventricular perfusion defect decreased significantly from 42% +/- 16% to 27% +/- 10% (group 1), whereas in the remaining 20 patients it showed no significant change (group 2). Predischarge low-dose dobutamine echocardiography showed a significant improvement in wall motion score index compared with baseline in group 1, from 1.62 +/- 0.28 to 1.41 +/- 0.24, P <.001, whereas in group 2 this index remained without significant change. Predischarge low-dose dobutamine echocardiography is an accurate tool for prediction of late recovery of myocardial perfusion after acute myocardial infarction treated with thrombolysis.
Surfactant secretion must be regulated to maintain a low surface tension in the lung during various conditions such as exercise. In vitro studies reveal a slow, unique exocytotic process at the interface of stimulated and constitutive exocytosis. The exocytotic mechanisms and sites of regulation in vivo, however, are still poorly understood.
Long-term, simultaneous, measurements of cytoplasmic free Ca(2+) concentrations and single exocytotic fusion events in surfactant-secreting type II cells were performed. All fusion (constitutive, phorbol ester-induced, and agonist-induced) was Ca(2+)-dependent. Kinetic analysis revealed that agonist (adenosine triphosphate [ATP])-induced fusion exhibited a kinetic pattern that correlated well with the Ca(2+) signal. The effects of Ca(2+) release from intracellular stores (early) and Ca(2+) entry (late) could be demonstrated for the first time by dissecting the slow (10-to-15-min) fusion response to ATP into these two components. Bath Ba(2+) or Sr(2+) could replace Ca(2+) to elicit a fusion response in thapsigargin-pretreated cells lacking ATP-induced Ca(2+) release from stores. Although the late response was partially inhibited by interrupting the phospholipase D-protein kinase C axis, a high Ca(2+) dependence of the entire secretory course was demonstrated by a significant correlation between the integrated Ca(2+) signal and the fusion response. There was also a highly significant correlation between constitutive and ATP-stimulated fusion activity in individual cells. We propose a common mechanistic model for all types of fusion in this slow secretory cell, in which constitutive and regulated forms of exocytosis are subject to the same principles of regulation.
The relationship between the dose of tacrolimus, trough tacrolimus blood concentration, and selected clinical endpoints (acute rejection, nephrotoxicity, and other toxicities) were examined in a prospective, multicenter clinical trial to validate the use of an enzyme-linked immunosorbent assay (ELISA) for monitoring whole-blood concentrations of tacrolimus in liver transplant patients. A total of 111 subjects from six transplant centers were evaluated over 12 weeks posttransplantation. In addition to trough tacrolimus blood concentrations, hematocrit, ALT, AST, GGTP, alkaline phosphatase, total bilirubin, serum creatinine, BUN, serum potassium, serum magnesium, blood glucose, and serum albumin were also measured. The relationship between trough tacrolimus blood concentrations and clinical endpoints was analyzed using both a logistic regression model and a Cox proportional hazard model. By logistic regression analysis, a statistically significant (p = 0.0465) relationship between increasing trough tacrolimus blood concentrations and decreasing risk of acute rejection was demonstrated over a 7-day time window. Nephrotoxicity and other toxicities also demonstrated statistically significant relationships with trough tacrolimus blood concentrations. The results of the Cox analysis were consistent with the logistic regression analysis. Using receiver operator characteristic curves, trough tacrolimus concentrations as measured by the ELISA method were able to differentiate the occurrence of nephrotoxicity and toxicity from nonevents. To minimize nephrotoxicity of tacrolimus, it is necessary to maintain trough blood concentrations below 15 ng/ml. This study demonstrates that the ELISA method used to measure tacrolimus blood concentrations in this study provides information of predictive value for managing the risk of nephrotoxicity, other toxicity, and rejection in liver transplant patients.
AIMS: To determine whether magnesium given orally decreases the recurrence rate of atrial fibrillation after elective direct current cardioversion of persistent atrial fibrillation. METHODS AND RESULTS: Consecutive outpatients were randomized to treatment with oral magnesium (10.3 mmol) or placebo twice daily in a double-blind fashion. Two groups were studied; magnesium study: 170 patients with atrial fibrillation persistent for >1 month, scheduled for their first direct current cardioversion. No concomitant antiarrhythmic drugs of class I or III were allowed. Sotalol and magnesium study: 131 patients with recurrence of persistent atrial fibrillation after previous direct current cardioversion, or a history of paroxysmal atrial fibrillation, treated with sotalol. Patients were followed until recurrence of atrial fibrillation or for at least 6 months. Magnesium study: at cardioversion 67 of 85 (79%) in the placebo group and 64 of 85 (75%) in the magnesium group had converted to sinus rhythm. At the end of the study, with a follow-up of 6 to 42 months, 15% of patients in the placebo group and 19% of patients in the magnesium group remained in sinus rhythm (Log rank test: P=0.37). Sotalol and magnesium study: pharmacological conversion to sinus rhythm, after oral treatment, was achieved in 34 of 131 (26%) patients. Sinus rhythm, with or without cardioversion, was restored in 89% and 85% of the patients in the placebo and magnesium groups, respectively. At the end of the study, with a follow-up of 6 to 42 months, 37% of patients in the placebo group and 30% of patients in the magnesium group remained in sinus rhythm (Log rank test: P=0.64). CONCLUSION: In patients with persistent atrial fibrillation, oral treatment with magnesium alone or as an adjuvant to sotalol, does not influence the recurrence rate of atrial fibrillation after elective cardioversion.
In western Canada, the Bt-10 resistance gene in wheat (Triticum aestivum) is effective against all the known races of common bunt caused by Tilletia tritici and T laevis. The genotypes of 199 F2 plants, originated from a cross between BW553 containing Bt-10 and the susceptible spring wheat cultivar 'Neepawa,' were established in greenhouse and field inoculation studies. A ratio of 1:2:1 resistant : heterozygous : susceptible was observed for bunt reaction, indicating that Bt-10 was expressed in a partially dominant fashion. A polymorphic DNA fragment, amplified using RAPD, and previously shown to be linked to Bt-10 was sequenced and SCAR (sequence characterized amplified region) primers devised. However, SCAR primers failed to amplify the polymorphic fragment. Restriction of PCR products with DraI revealed a polymorphic fragment of 490 bp resulting from a single base pair difference between lines possessing Bt-10 and those lacking the gene. As per the base pair difference, FSD and RSA primers were designed to generate a 275-bp polymorphic DNA fragment. Both 275- and 490-bp polymorphic fragments were present in all of the 22 cultivars known to carry Bt-10, and absent in all 16 cultivars lacking Bt-10. A 3:1 ratio was observed for presence: absence of the 275-bp marker in the F2 population. Using Southern analysis, the 490-bp fragment was effective in differentiating homozygous resistant plants from those heterozygous for Bt-10, based on its presence and the hybridization signal strength. A 1:2:1 resistant : heterozygous : susceptible ratio was also observed for the molecular marker and corresponded to 88% of the phenotypes deduced from the original F2 population. The molecular marker was estimated to be between 1.1 cM and 6.5 cM away from the Bt-10 resistance gene, based on the segregation analysis. Segregation analyses of Bt-10 and the 275-bp marker, evaluated in three different Canada Prairie Spring (CPS) wheat populations, demonstrated a segregation ratio of 3:1 for the molecular marker in two of the populations. These results demonstrated that the PCR marker system using the FSD and RSA primer pair permitted a rapid and reliable identification of individual lines carrying the Bt-10 gene for resistance to common bunt.
In patients with atrial fibrillation, the reduced right ventricular function determined by tricuspid annular motion before cardioversion returns to normal 1 month after successful cardioversion to sinus rhythm. The simplicity of recording the tricuspid annular motion provides an easy opportunity to assess right ventricular function following electroconversion of atrial fibrillation to sinus rhythm.
The present double-blind, placebo-controlled study investigated the effects of intravenous magnesium on heart rate and rate variability in 30 patients with chronic atrial fibrillation. During standardized conditions, intraindividual variation in heart rate and rate variability was low in patients with chronic atrial fibrillation and magnesium had no effect on heart rate or rate variability.
We retrospectively analyzed factors influencing PBPC mobilization during steady-state hematopoiesis in 52 patients with malignant lymphoma (n=35) or multiple myeloma (n=17) who received 77 cycles of G-CSF (12.5-50 microg G-CSF/kg/day). For 15 of these patients, the first mobilization cycle (12.5 microg G-CSF/kg/day) was followed by a second course with an increased dose of G-CSF (25 or 50 microg/kg/day). Leukapheresis was started on day 4, about 2 h after s.c. G-CSF administration, and repeated on 2-5 consecutive days. CD34+ cells were determined by flow cytometry in each apheresis product and in the peripheral blood prior to G-CSF administration, beginning on day 4. Colony assays were performed on cryopreserved samples prior to autografting. In the 15 patients receiving two mobilization cycles the higher G-CSF dose was associated with higher levels of CD34+ cells, a higher mean yield of CD34+ cells per apheresis (p<0.05), and a higher percentage of successful (>2x10(6) CD34+ cells/kg) collections (p=0.058). Patients with limited previous cytotoxic therapy (n=19, up to six cycles of a standard regimen such as CHOP and/or less than 20% marrow irradiation) who received a daily dose of 12.5 microg G-CSF/kg had higher levels of circulating CD34+ cells, a higher mean yield of CD34+ cells per apheresis (p<0.05), and a higher percentage of successful collections (p<0.05) compared with patients previously treated with more intensive radiochemotherapy (n=15). Ten of 20 patients (50%) who failed during the first cycle were successful during subsequent cycles with escalated doses of G-CSF. Trough levels of circulating CD34+ cells on day 4 were predictive for success or failure to achieve >2x10(6) CD34+ cells/kg, especially in heavily pretreated patients. In conclusion, a daily dose of 12.5 microg G-CSF/kg seems sufficient to mobilize PBPC during steady-state hematopoiesis in the majority of patients who have received limited previous radiochemotherapy. Higher doses of G-CSF, up to 50 microg/kg/day, mobilize more PBPC and should be considered for patients previously treated with intensive radiochemotherapy or those failing to mobilize sufficient numbers of CD34+ cells with lower doses of G-CSF.
BACKGROUND: This study was undertaken to evaluate the ability of predischarge low-dose dobutamine echocardiography to predict late left ventricular functional recovery after thrombolyzed acute myocardial infarction. METHODS AND RESULTS: Low-dose dobutamine echocardiography was performed in 54 patients 4 +/- 2 days after acute myocardial infarction treated with thrombolysis. Follow-up resting echocardiography was carried out in 49 of these patients at a mean of 18 +/- 6 months later. Viability was defined as recovery of myocardial function at follow-up, expressed as an improvement of wall motion of at least 1 grade or more in at least 2 contiguous infarct zone segments. In 24 of the 49 patients (group I), wall motion at follow-up improved in comparison with the early resting echocardiographic study (1.72 +/- 0.29 vs 1.37 +/- 0.34, P <.001). In the remaining 25 patients (group II), no wall motion enhancement was seen at follow-up (1.57 +/- 0.38 vs 1.58 +/- 0.36, NS). In 22 of the 24 patients in group I, early low-dose dobutamine echocardiography showed improvement in the wall motion score index compared with baseline resting measurements (1.72 +/- 0.29 vs 1.44 +/- 0.24, P <.001). The positive and negative predictive value of early low-dose dobutamine echocardiography to predict functional recovery was 76% and 92%, respectively. CONCLUSION: Predischarge low-dose dobutamine echocardiography is an accurate tool for detecting viable myocardium and predicting late left ventricular recovery after acute myocardial infarction treated with thrombolysis.
Pulmonary surfactant is secreted via exocytosis of lamellar bodies (LBs) by alveolar type II cells. Here we analyzed the dependence of LB exocytosis on intracellular Ca(2+) concentration ([Ca(2+)](i)). In fura 2-loaded cells, [Ca(2+)](i) was selectively elevated by flash photolysis of a cell-permeant caged Ca(2+) compound (o-nitrophenyl EGTA-AM) or by gradually enhancing cellular Ca(2+) influx. Simultaneously, surfactant secretion by single cells was analyzed with the fluorescent dye FM 1-43, enabling detection of exocytotic events with a high temporal resolution (T. Haller, J. Ortmayr, F. Friedrich, H. Volkl, and P. Dietl. Proc. Natl. Acad. Sci. USA 95: 1579-1584, 1998). Exocytosis was initiated at a threshold concentration near 320 nmol/l with both instantaneous or gradual [Ca(2+)](i) elevations. The exocytotic response to flash photolysis was highest during the first minute after the rise in [Ca(2+)](i) and thus almost identical to purinoceptor stimulation by ATP. Correspondingly, the effects of ATP on initial secretion could be sufficiently explained by its ability to mobilize Ca(2+). This was further demonstrated by the fact that exocytosis is significantly blocked by suppression of the ATP-induced Ca(2+) signal below approximately 300 nmol/l. Our results suggest a highly Ca(2+)-sensitive step in LB exocytosis.