[Inactivation of AMP-deaminase from white muscle of Cyprinus carpio in the systems with free radical oxidation].

The purification and in vitro inactivation of AMP-deaminase from white muscle of carp Cyprinus carpio were conducted in the Fe2+/H2O2 and Fe2+/ascorbate oxidation systems. The enzyme activity decreases by 50% within 30 minutes of incubation in the presence of 100 microM of hydrogen peroxide and 5 microM of ferrous sulfate. Inactivation depended on incubation time and concentrations of FeSO4 and H2O2. In the system Fe2+/ascorbate the enzyme activity decreased by 50% at concentration of ascorbate 1 mM and 5 ferrous sulfate microM. Sodium nitrite did not affect the activity. S(0.5) and n(H) of both native and partially inactivated enzymes were virtually the same, while maximal activity of the inactivated enzyme was 2-3-fold lower than that of the native one.

The accuracy of myocardial perfusion SPECT imaging in the evaluation of coronary artery disease in women and men.

BACKGROUND: The purpose of this study is to compare sensitivity, specificity and accuracy of myocardial perfusion SPECT for the detection of coronary artery disease (CAD) in women and men. MATERIAL AND METHODS: 588 patients (455 males and 133 females, 273 after a previous myocardial infarction) underwent stress myocardial perfusion SPECT. The accuracy of myocardial perfusion SPECT was proved by coronary angiography (stenosis > 50% was considered as a CAD). RESULTS: The sensitivity of SPECT was slightly higher, but statistically not significant, in men than in women (94% versus 91%, p > 0.05). The specificity was higher in women than in men (93% versus 82%), but this difference was not statistically significant either (p > 0.05). The accuracy of SPECT was the same for both sexes (92%). In angiographically verified group of patients the selection bias was obvious--patients with CAD dominated (74%) and the fraction of patients with CAD in men's group (83%) was significantly higher than in women's group (50%), p < 0.05. CONCLUSIONS: No significant difference was revealed in the accuracy of myocardial perfusion SPECT in men and women. Our results are in accordance with the prevailing opinion in literature that discovered differences in sensitivity, specificity and diagnostic accuracy are usually not statistically significant or that they can be explained by the selection bias of patients in angiographically verified groups (significantly higher fraction of patients with CAD in men's group).

[Prognostic significance of stress tomographic scintigraphy of myocardial perfusion in diabetic patients]. / Prognostický význam zátezové tomografické scintigrafie myokardiální perfuze u diabetických pacientu.

OBJECTIVE: To assess the prognostic value of SPECT in patients with diabetes (DM) and without DM. METHOD: A total of 366 patients (104 women, 262 men) were examined by T1 +/- 201 or Tc-99m-MIBI SPECT of the myocardium. DM was recorded in 149 patients, 217 patients did not suffer from DM. The SPECT findings were classified as normal and abnormal perfusion (fixed or reversible perfusion defect). A serious cardiac event was defined as sudden cardiac death or non-fatal myocardial infarction. Also angina pectoris requiring revascularization was recorded. RESULTS: During the average investigation period of 24 +/- 7 months we recorded in 147 patients with a normal load perfusion only one non-fatal myocardial infarction and the risk of a serious cardiac event was low: 0.3% per annum in the whole group. A significantly higher incidence of a severe cardiac event, 5.9% per year, was recorded in 219 patients with an abnormal finding on SPECT (10 deaths and 16 non-fatal myocardial infarctions, P < 0.01). On comparison of the frequency of serious cardiac events in groups of patients with and without DM no significant difference was found as regards normal load perfusion of the heart muscle (1.1% vs. 0%, P = NS) even in patients with an abnormal SPECT finding (5.8% vs. 6.1, P = NS). CONCLUSION: SPECT myocardial perfusion makes it possible to stratify the risk in patients with DM similarly as proved previously in non-selected groups. Similarly as in patients without DM normal perfusion in diabetic patients predicts a benign prognosis.

Prognostic value of myocardial perfusion tomographic imaging in patients after percutaneous transluminal coronary angioplasty.

PURPOSE: The authors assessed the prognostic value of stress myocardial perfusion tomographic imaging (SPECT) in patients with recurrent angina or inconclusive results of exercise electrocardiographic tests after successful percutaneous transluminal coronary angioplasty (PTCA). MATERIALS AND METHODS: After PTCA, 70 patients (54 men, 16 women; 41 after myocardial infarction; mean age, 56 +/- 9 years) underwent TI-201 or Tc-99m sestamibi SPECT studies. SPECT patterns were divided into normal (n = 25), fixed defects (n = 15), and reversible or combined fixed plus reversible defects (n = 30). A cardiac event was defined as either cardiac death, nonfatal myocardial infarction, or unstable angina requiring further revascularization. RESULTS: During an average follow-up of 25 +/- 10 months, two patients had severe outcomes (one cardiac death and one nonfatal myocardial infarction), and revascularization was required in 13 patients. In patients with normal SPECT or fixed defects, the annual event rate was low (1.2%), with only one revascularization. In patients with reversible or combined defects, the annual event rate was significantly greater (22.4%; chi square = 17.32, P = 0.00003). CONCLUSIONS: Normal perfusion or fixed defects predict a benign prognosis in patients after successful PTCA. The presence of stress-induced reversible defects appears to be the best predictor of future cardiac events.

Medical radiation exposure of the Czech Republic paediatric population due to diagnostic nuclear medicine.

BACKGROUND: The information on the results of the survey of collective effective dose SE to children in the Czech Republic (CR) due to nuclear medicine examinations performed during the one-year period July 1995-June 1996. METHODS: The estimate of SE was based on the annual frequency of nuclear medicine procedures and the type and amount of administered radiopharmaceuticals provided by the General Health Insurance Company covering 75% of the CR population. RESULTS: In the period considered, 15,281 examinations of children younger than 18 years were performed, which represented 7.5% of total nuclear medicine examinations in the CR. The annual SE 43.8 man Sv formed 5.1% of the collective effective dose to all patients examined by nuclear medicine procedures in the CR. The contributions of procedures in various body systems of children to the annual SE: urinary tract 32.8%, bone 30.4%, brain 12.8 %, cardiovascular 6.4%, GIT 6.0%, thyroid 4.4%, lungs 3.0%, other 4.2%. In comparison with a similar survey performed in the CR in 1987 (based on a questionnaire) there was an increase in SE to children younger than 15 years by a factor of 2.8, mainly due to the expansion of kidney and bone investigations. CONCLUSIONS: The mean effective dose per exam to children younger than 15 years was 60% of that for adults, which was in agreement with the requirement that the radiation burden of children should not exceed that of adults. The use of data from the Insurance Company for regular conducting of such surveys appeared to be not as cumbersome and time-consuming as questionnaire surveys.

Optimizing fluorescent labeling of endothelial cells for tracking during long-term studies of autologous transplantation.

BACKGROUND: The fluorescent marker PKH26 has been demonstrated to be useful for the tracking of endothelial cells in short-term studies; however, the optimal labeling conditions for long-term implants have not been determined. This study was designed to evaluate the effects of PKH26 on endothelial cell proliferation and to identify labeling conditions that would yield the greatest fluorescence over time without adversely affecting cell viability. MATERIALS AND METHODS: Canine jugular vein endothelial cells (CJVECs) were labeled with 0. 04 microM PKH26. Proliferation of labeled and control cells was assessed for 8 consecutive days by [(3)H]thymidine uptake. In a second experiment, CJVECs were labeled at concentrations of 0, 5, 8, 10, and 20 micromol/L. Cells were maintained in culture for 60 days. The fluorescence intensity of each cell population was measured using two techniques. At baseline and at 60 days, fluorescence was measured using a fluorescence-activated cell sorter. On days 14, 28, 45, and 60 fluorescence was measured by constructing gray-scale histograms from photomicrographs taken of each flask under rhodamine illumination. Mean viable cell number for each concentration was determined after 60 days. RESULTS: In the first experiment, PKH26-labeled and unlabeled CJVECs demonstrated nearly identical growth curves, suggesting that PKH26 had no adverse effect on proliferation. In the second experiment, after 60 days, the 10 and 20 microM groups displayed greater fluorescence by histogram than the 0, 5, or 8 microM groups; however, they were not significantly different from each other (mean intensity 8.2 vs 9.1, P > 0.05, Student-Newman-Keuls test for multiple comparisons). Over 60 days, the cells labeled with 20 microM PKH26 experienced the only significant decrease in viable cells compared to the unlabeled group (5.5 x 10(5) vs 9.6 x 10(5) cells/flask, P < 0.05). Importantly, we observed no significant differences in cell number between the 10 microM group and the lower concentrations compared to the unlabeled cells (P > 0.05). CONCLUSIONS: We conclude that a concentration of 10 microM PKH26 provides the optimal labeling condition for endothelial cells when long-term tracking is desired.

[Use of radiopharmaceuticals, radiation burden and radiation risks in examination procedures in nuclear medicine]. / Aplikované aktivity radiofarmak, radiacní zátez a radiacní riziko vysetrovacích postupu v nukleární medicíne.

Consistent with the worldwide development of nuclear medicine, in the Czech Republic in 1987 to 1996 partly the spectrum of used radiopharmaceutical preparations and their ratio changed. In the majority of radiopharmaceutical preparations the mean administered activity increased by 20 to 80% and the range of activities administered in different departments diminished somewhat. Adherence to principles of radiation protection of patients is assisted by guidelines of administered activities of radiopharmaceutical preparations laid down in new regulations on radiation protection valid in the CR from the middle of 1997. The radiation burden associated with the majority of examinations in nuclear medicine expressed as the effective dose is comparable with the radiation burden of radiodiagnostic examinations, only after administration of preparations with 131I, 201Tl, 67Ga and 111In it is markedly higher. Based on knowledge of the effective dose it may be concluded that the lifetime extra risk of a fatal tumour due to administration of radiopharmaceutical preparations is by two to three orders lower than the lifetime risk of spontaneous development of fatal tumours. A special feature of radiation risk is its hypothetical character--it is frequently projected into the distant future of human life, contrary to immediate non-radiation risks of some medical procedures and risks in the living environment which frequently are higher. Any examination using ionising radiation can be made only if the expected health benefit for the patient is significantly greater than the radiation risk. Excessive fear of radiation risk should not lead to refusal of justified examinations with possible subsequent serious health damage for the patient.

Hepatic perfusion changes in patients with cirrhosis indices of hepatic arterial blood flow.

PURPOSE: This study used radionuclide angiography to evaluate semiquantitatively the hepatic arterial blood flow changes associated with cirrhosis. METHODS: The parameters of net arterial hepatic perfusion were estimated by analysis of first-pass flow studies in 11 control participants and in 15 patients with cirrhosis (Child-Pugh classification B-C). Hepatic, renal, and splenic time-activity curves were generated, normalized per pixel, and corrected for background. The rate of hepatic arterial blood flow was compared with the reference kidney and spleen perfusion using the hepatorenal and hepatolienal perfusion indices, respectively. These indices were defined as: PI = area under hepatic curve limited by time of the renal (splenic) curve peak/area under renal (splenic) curve to its peak RESULTS: The values of both these perfusion indices are significantly greater in the patients with cirrhosis than in controls (hepatorenal perfusion index, P < 0.01; hepatolienal perfusion index, P < 0.05). The values of the hepatic perfusion index (the ratio of the arterial to the total liver blood flow), which were also calculated, were elevated in the patients with cirrhosis (P < 0.01). CONCLUSIONS: The results confirm that the net hepatic arterial blood flow is increased in patients with cirrhosis. Radionuclide angiography accompanied by calculation of arterial perfusion indices may provide semiquantitative parameters of net hepatic arterial blood flow.

[Myocardial perfusion tomography imaging in patients without prior myocardial infarction--comparison between 201Tl, 99mTc-tetrofosmin and dual-isotope protocol (rest 201Tl/stress 99mTc-tetrofosmin)]. / Tomografické zobrazení myokardiální perfuze u pacientu bez infarktu myokardu--srovnání 201Tl s 99mTc-tetrofosminem a dual-izotopovým protokolem (klid 201Tl/zátez 99mTc-tetrofosmin).

The authors compared retrospectively the diagnostic accuracy of three protocols--201Tl stress-redistribution, 2-day 99mTc-tetrofosmin and the dual-isotope protocol (rest 201Tl/stress99m Tc-tetrofosmin)--for the detection of ischaemic heart disease (IHD) in a total of 115 patients without previous myocardial infarction (IM). 201Tl protocol (group A) was used to examine 43 patients, incl. 15 women, mean age 51 years (38-69 years). The 2-day 99m Tc-tetrofosmin protocol was used to examine 39 patients incl. 12 women, mean age 49 years (30-71 years). The dual-isotope protocol was used in 33 patients, incl. 8 women, mean age 52 years (31-69 years). In all patients coronarography was performed, stenosis of the artery >50% was considered significant for IHD. The sensitivity of the dual-isotope protocol was 95% and 96% resp. and 92% in the 2-day tetrofosmin and 201Tl protocol, (p>0.05). The specificity of the dual-isotope protocol was 82% and 89% resp. and 88% in the 201Tl and 2-day tetrofosmin protocol, (p >0.05). The diagnostic accuracy was 91% in the dualisotope and 201Tl protocol and 92% in the 2-day protocol (p>0.05). No significant difference was revealed in the sensitivity, specificity and diagnostic accuracy for the detection of IHD between different protocols in patients without previous IM.

[Correlation of SPECT myocardial stress scintigraphy with coronary angiography]. / Korelace zátezové perfuzní scintigrafie myokardu SPECT s koronarografickými nálezy.

The sensitivity and specificity of a myocardial perfusion single-photon emission tomography (SPECT) for the detection of the coronary artery disease (CAD) was investigated. As a gold standard the coronary angiography was used--the stenosis of the artery by 50% and more was considered as significant for CAD. After performing SPECT studies patients were examined by coronary angiography according to the attending physician's judgement. In the retrospectively created group of 123 subjects (94 males and 29 females, mean age 53 years, range 30-69 years) 55 patients underwent myocardial infarction (MI). The stress was performed using a bicycle ergometer or, in case of patient's inability, an intravenous dipyridamole infusion. Before december 1995 201T1 was used in all patients, after this date only in patients with prior MI with regard to the advantage of this radionuclide for the detection of the ischemic but viable myocardium. Other patients were administered 99mTc-tetrofosmin. In our group of patients we found the high sensitivity of myocardial SPECT of 94.5% 86 (from 91), specificity of 84.4% (27 from 32) corresponding to usually values and the high diagnostic accuracy of 91.9% (113 from 123). Specificity can be impaired by marked attentuation of gamma-rays by overlying soft-tissue structures such as the breast or diaphragm. The artificial imaging of the inferior wall due to the infradiaphragmatic structures represents the most serious problem-there were four false positive findings in our group; further ones in our 99mTc-tetrofosmin studies were avoided by the routine use of the prone position recommended in the literature. The validity of SPECT myocardial perfusion findings should be verified by coronary angiography; however, one must be aware of some questionable points inherent in this comparison.

[Experimental-morphological and clinical basis of preventive methods for the intraoperative complications of the amputation of the lower extremity at the thigh level]. / Eksperimental'no-morfolohichne ta klinichne obhruntuvannia sposobiv profilaktyky uskladnen' pry amputatsii kintsivky na rivni stehna.

The nonexpediency of the tourniquet and retractor application while the extremity amputation conduction on the femur level, the necessity in renunciation of the neurotomy conduction using an amputational knife, scalpel or scissors were proved, basing on the topographic-anatomic (53 cadavers), experimental (130 white rats and 50 dogs) and clinical (40 patients) data.

Fibrin glue containing fibroblast growth factor type 1 and heparin decreases platelet deposition.

BACKGROUND: The early success rates of endarterectomy and angioplasty are influenced by the thrombogenicity of the deendothelialized surface. We previously reported decreased platelet deposition after 30 and 120 minutes and after 28 days on expanded polytetrafluoroethylene (ePTFE) grafts coated with fibrin glue (FG) containing fibroblast growth factor type 1 (FGF-1) and heparin in canine aortoiliac bypass grafts when compared with control uncoated grafts. The FG/FGF-1/heparin coating has been shown to enhance spontaneous endothelialization at 28 days in canine ePTFE bypass grafts. The current study evaluates the thrombogenicity of this FG/FGF-1/heparin suspension applied to a balloon de-endothelialization model of endarterectomy in canine carotid arteries. METHODS: Nine dogs underwent bilateral, deendothelialization balloon injury to 6-cm segments of their carotid arteries. Fibrin glue (fibrinogen 32.1 mg/mL + thrombin 0.32 U/mL) containing FGF-1 (11 ng/mL) and heparin (250 U/mL) was applied to the luminal surface of one carotid artery in each dog. Both femoral arteries were circumferentially dissected but not balloon injured; one femoral artery was clamped for the same period as the carotid arteries. In the 6 acute dogs, 10 minutes prior to the restitution of flow in both carotid arteries and one femoral artery, 4 to 8 x 10(9) (111)In-labelled autologous platelets were injected intravenously. Four-cm segments of both carotid and femoral arteries were excised after 15 or 120 minutes of circulation (n = 3/time/artery, 24 arteries). In the 3 chronic dogs, the radiolabelled platelets were injected 30 days after carotid injury. The carotid and femoral vessels were then excised after 120 minutes of perfusion. Radioactive platelet deposition was quantitated by gamma counting. RESULTS: After 2 hours, the injured carotid arteries demonstrated significantly more platelet deposition than either uninjured femoral artery controls (P < 0.001). There was also a significant 45.2% decrease (P = 0.008) in platelet deposition on the balloon injured carotid arteries treated with FG/FGF-1/heparin when compared with balloon injured carotid arteries alone. At 30 days there was an insignificant trend toward decreased thrombogenicity in the FG/FGF-1/heparin treated injured carotids. CONCLUSION: Surface coating with FG/FGF-1/heparin significantly decreases platelet deposition on balloon injured canine carotid arteries after 2 hours of perfusion and may be clinically applicable in endarterectomy and angioplasty procedures. The long-term induction of reendothelialization of arterial surfaces by this technique is under investigation.

Fibrin glue containing fibroblast growth factor type 1 and heparin with autologous endothelial cells reduces intimal hyperplasia in a canine carotid artery balloon injury model.

PURPOSE: Intimal hyperplasia plagues all types of vascular intervention. Early confluent re-endothelialization may attenuate the smooth muscle cell (SMC) proliferative response. We previously reported that fibroblast growth factor type 1 (FGF-1) and heparin at relative concentrations of 10 ng/ml:250 U/ml delivered in a fibrin glue (FG) suspension can selectively stimulate endothelial cells (EC) and inhibit SMC proliferation in cell culture. This current study evaluates this surface treatment with and without seeded autologous ECs on intimal hyperplasia in a canine carotid artery balloon injury model. METHODS: Twenty-nine adult dogs underwent bilateral balloon injury to a 6 cm segment of their carotid arteries. The injury resulted in a reproducible removal of the intima and 4 to 6 medial lamellae. Nine dogs were used in part I to determine the percent retention of FGF-1 and EC when applied in a FG suspension to the balloon-injured carotid arteries. Part 2 used the remaining 20 dogs to determine the effect of this surface treatment on intimal hyperplasia. In 10 group I dogs, FG (fibrinogen 32.1 mg/ml and thrombin 0.32 U/ml) containing FGF-1 (11 ng/ml) and heparin (250 U/ml) was applied to the luminal surface of one carotid artery, whereas the contralateral carotid artery underwent balloon injury alone. In 10 group II dogs, an identical FG preparation with FGF-1 and heparin was applied to the surface of one carotid artery, whereas the contralateral carotid artery received FG/FGF-1/heparin that also contained autologous ECs (P3; 5 x 10(4) to 10 x 10(4) cells/cm2). Five dogs from both group I and group II were killed at 10 days and the remaining 10 dogs at 30 days. Histologic analysis and computerized morphometric analysis were used to determine intimal and medial thickness and area, percent endothelialization, and medial SMC proliferative rate. RESULTS: There was no measurable neointima in any 10-day dog. There was no difference in neointimal area between the treatments in group I 30-day dogs. There was a significant decrease in maximal neointimal area, intima/media thickness ratio, and intima/media area ratio in group II 30-day dogs that were treated with FG/FGF-1/heparin plus EC. There was an insignificant increase in percent EC coverage and an insignificant decrease in medial SMC proliferative rate in group II 10-day dogs treated with FG/FGF-1/heparin plus EC. CONCLUSIONS: In this canine carotid model, FG with FGF-1 and heparin did not induce significant intimal or medial thickening after 10 or 30 days when compared with vessels that were only balloon-injured. The seeding of autologous ECs within the FG/FGF-1/heparin suspension caused a reduction in neointima formation with no concomitant medial thickening 30 days after injury. The use of FG to locally deliver FGF-1 and ECs may have clinical relevance in the inhibition of intimal hyperplasia.

Absolute quantitation of gallium-67 citrate accumulation in the lungs and its importance for the evaluation of disease activity in pulmonary sarcoidosis.

Our modification of a method for the absolute quantification of gallium-67 uptake in lungs with a scintillation camera and computer is described. The uptake of 67Ga in lungs, expressed in percentage of administered radioactivity, was determined by the transmission-emission method. We proved theoretically and experimentally that a 67Ga planar source could be replaced with a 57Co planar source. The performance of lung perfusion scans allows a more accurate delineation of the regions of interest on gallium scans. The method was applied to control subjects (n = 27) and to patients (n = 114) suffering from biopsy-proven pulmonary sarcoidosis (28 with inactive and 86 with active disease). The obtained results were compared with chest X-ray findings, the percentage of lymphocytes in the bronchoalveolar fluid (BAF-ly%), and serum angiotensin-converting enzyme (SACE) values. The method seems suitable for the assessment of disease activity in sarcoidosis. It is more accurate in detecting parenchymal involvement in lung sarcoidosis than the commonly used X-ray criteria. No correlation was found between 67Ga uptake and the BAF-ly% and SACE values.

[Activity of administered radiopharmaceutical agents and the radiation burden of the Czechoslovak population caused by nuclear medicine]. / Aktivity aplikovaných radiofarmak a radiacní zátez obyvatelstva CSFR z nukleární medicíny.

The authors assessed by means of questionnaires the activities of radiopharmaceuticals administered in departments of nuclear medicine in Czechoslovakia. The mean activities of individual radiopharmaceuticals are roughly equal as in Great Britain, but lower than in the Canadian province of Manitoba. The differences of activities used in different departments are approximately equal in all compared countries. In the Czech Republic the annual collective effective dose equivalent from nuclear medicine was 433 Sv in 1983 and 609 Sv in 1987. The mean effective dose equivalent per examination was 2.23 mSv in 1983 and 2.44 mSv in 1987. The mean effective dose equivalent per inhabitant of the Czech Republic was 0.042 mSv in 1983 and 0.059 mSv in 1987. The radiation dose of the Czech population from nuclear medicine amounts approximately to one tenth of the load from radiodiagnostics.