Studies on the labeling of streptokinase with 99mTc for use as a radiopharmaceutical in the detection of deep-vein thrombosis: concise communication.

Streptokinase was labeled with 99mTc using both stannous chloride and stannous pyrophosphate as reducing agents. Sixty to seventy-five percent of the 99m Tc was incorporated into streptokinase using stannous chloride as a reducing agent at pH 1-2, wheras 50-60% was incorporated using stannous pyrophosphate at neutral pH. Increasing the pH from 2 to 7 in the presence of stannous chloride caused the release of 15-20% of the protein-bound 99mTc. Incorporation of 99mTc into protein was relatively slow: labeling required 2-3 hr at room temperature. The concentration of stannous pyrophosphate required for optimum labeling varied between 10(-5) and 10(-2) M. Polyacrylamide-gel electrophoresis showed that the filler substance in commercial streptokinase was also labeled with 99mTc. However pure streptokinase gave a homogenous protein band after polyacrylamide-gel electrophoresis. This protein band coincided with the peak of streptokinase-bound 99mTc. The results obtained may partially explain why 99mTc-labeled streptokinase lacks the necessary specificity for the satisfactory location of blood clots in vivo.

Accumulation of technetium-99m stannous pyrophosphate in contused myocardium.

The uptake of technetium-99m stannous pyrophosphate in contused myocardium was measured as a function of time from the insult. The free wall of the dog's left ventricle was surgically exposed and struck with a spring-loaded paddle. Pyrophosphate was injected intravenously from 1 1/2 to 47 1/2 hr after the injury. After 1/2 hr of incubation the hearts were removed and the Tc-99m content of contused and noncontused myocardium was measured. Pyrophosphate was concentrated in contused myocardium at all of the time periods tested. Contused-to-normal ratios for pyrophosphate uptake ranged from 8.1 (8 hr) to 41.9 (48 hr).

A comparison of radiotracer and biochemical methods for the quantitation of experimental myocardial infarct weight: in vitro relationships.

Anterior myocardial infarction (MI) was produced in conscious dogs to evaluate the relationships among: a) cardiac technetium-99m stannous pyrophosphate (TcPPi) accretion, b) creatine phosphokinase (CPK) depletion, and c) postmortem MI weight, infarct structure, and histology. In vitro, there was a close relationship between measured MI weight and MI weight calculated by the TcPPi accretion (r = 0.96) or CPK depletion (r = 0.93) in representative "cross-sectional" MI samples. Cardiac TcPPi accretion and CPK depletion showed a curvilinear relationship over the spectrum of tissue samples. Adjacent to infarcts, there was marked TcPPi uptake and modest CPK depletion where histology suggested ischemia without infarction. Within infarcts, microscopically visible calcium was rare in this series, suggesting little intracellular calcium accumulation, insensitivity of the von Kossa staining technique, and/or other cellular mechanisms to account for Tc-PPi uptake in this conscious dog model without reperfusion.

Myocardial uptake of technetium-99m stannous pyrophosphate in experimental viral myopericarditis.

Distribution of technetium-99m stannous pyrophosphate was studied in mice with experimentally induced viral myopericarditis. Myocardial and bone uptakes of Tc-PPi were compared in 55 mice inoculated with coxsackievirus B3 (Nancy strain). The myocardium-to-bone uptake ratio in 33 mice with myopericarditis was increased to a greater extent than that seen in 22 mice without myopericarditis (p less than 0.001). In the severely involved heart, the uptake per gram exceeded that in the bone. Myocardial uptake in myopericarditis can be visualized on a whole-body image using a pinhole collimator and a left lateral view. Our experimental studies suggest the potential clinical usefulness of myocardial scintigraphy in viral myopericarditis.

Quantitation of skeletal-muscle necrosis in a model compartment syndrome.

UNLABELLED: Skeletal-muscle necrosis was evaluated in previously pressurized canine compartments using technetium-99m stannous pyrophosphate and classic histological criteria. Intracompartmental necrosis was quantitated in the anterolateral muscle compartment of each dog by uptake of 99mTc stannous pyrophosphate using the contralateral anterolateral compartment as an internal control. Representative specimens of muscle were sampled in experimental and control legs of each dog and were analyzed by qualitative histological techniques. Muscle necrosis was assessed in compartments forty-eight hours after pressurization to levels of ten to 120 millimeters of mercury for eight hours in thirty-seven dogs. In another dog, neither anterolateral compartment was pressurized so that both compartments acted as control muscle. The results in these experiments identify a threshold pressure level (thirty millimeters of mercury) and duration (eight hours) at which significant muscle necrosis occurs at normal blood pressure. Our findings imply that a quantitative relationship exists between incorporation of 99mTc stannous pyrophosphate and the level of intracompartmental pressure. This uptake technique, however, is not suitable for diagnosing compartment syndrome in patients with a threatened compartment syndrome. We suggest that intracompartmental pressure measurements by the wick-catheter technique, in conjunction with clinical findings, offer the best means for diagnosing compartment syndrome. CLINICAL RELEVANCE: Significant muscle necrosis associated with an impending compartment syndrome occurs at a threshold intracompartmental pressure of thirty millimeters of mercury after eight hours. Since time variables are often unknown in suspected compartment syndromes, fasciotomy is recommended when intracompartmental pressure exceeds thirty millimeters of mercury in a patient with normal blood pressure. The use of this threshold pressure level as an indication for fasciotomy requires a device for measuring intracompartmental pressure such as the wick catheter.

Autoradiographic comparison of 96Tc-pyrophosphate and 45Ca bone uptake.

To compare the skeletal distribution of calcium and pyrophosphate complexes, autoradiograms of the growing zone of 10-week-old rabbit femurs were performed. Due to its stability to chemical and enzymatic hydrolysis and to the relatively long half-life of the isotope, 96Tc-labelled pyrophosphate was used. Calcium uptake was studied with carrier free 45Ca. For both tracers, the blood supply plays a significant role by delivering the radiopharmaceuticals. In a second step their partition was governed by mixing in different bone structures: calcium was incorporated inside the bone trabeculae and the mineral mass whereas labelled pyrophosphate remained in linear patterns surrounding bone spicules.

Comparative quality control of 99mTc-Pyrophosphate and 99mTc-diphosphonate radiopharmaceuticals.

The results of analysis of 99mTc-Pyrophosphate (99mPyP), taken as a representative of the group of compounds having an organic P-O-P bond, and of the three diphosphonate compounds: methylene diphosphonate (MDP), 2,3-dicarboxy propane diphosphonate (DPD) and ethane-1-hydroxy-1, 1-diphosphonate (EHDP), which differ in their chemical structure, are shown. Also, some physicochemical parameters such as chloroform-water apparent partition coefficient, the osmotic pressure and pH values in final preparations were studied. The radiochemical purity of these radiopharmaceuticals was determined by the two methods: Sephadex chromatography for separation of 99mTc-hydrolysate and TLC on silica gel with 85% methanol for the determination of free 99mTcO-4. The yield of labelling for both methods was over 90%. Also, pharmacokinetic parameters such as binding to the plasma proteins and to erythrocytes were determined. 99mTc-PyP binding to plasma proteins was higher than the binding of diphosphonate compounds. The quantitative distribution of preparations was determined in experimental animals.

Myocsrdial uptake of technetium-99m stannous pyrophosphate following direct current transthoracic countershock.

The effect of direct current (DC) countershock upon myocardial technetium-99m stannous pyrophosphate (PYP) uptake was studied in 22 dogs. All eight dogs imaged had positive abnormal PYP scintigrams that were usually indistinguishable from experimental infarction. In three animals, additional areas of radionuclide uptake were seen in overlying noncardiac tissue. Left and right ventricular myocardial PYP uptake averaged (+/- SEM) 23 +/- 5 times control and 24 +/- 6 times control, respectively. These activity ratios occurred without reduction in regional myocardial blood flow (RMBF), and were associated with histologic evidence of necrosis. The necrosis was usually epicardial, corresponding to the transmural site of greatest PYP uptake. The magnitude of PYP accumulation and the weight of damaged tissue increased with increasing applied energy. Thus, PYP uptake following DC countershock could result in false-positive interpretation of acute ischemic myocardial infarction. Since RMBF is normal in regions of PYP uptake, the major determinant of radionuclide accumulation is the extent of cellular damage.

Effect of methylprednisolone upon technetium-99m pyrophosphate assessment myocardial necrosis in the canine countershock model.

Repeat DC countershock reproducibly results in myocardial necrosis in dogs. In this model, myocardial technetium-99m pyrophosphate (PYP) uptake correlates linearly with tissue creatine kinase depletion (r = -0.83). The effect of pretreatment with methylprednisolone (MP) was studied with PYP in 25 dogs. In myocardium damaged by countershock, 12 MP dogs had higher tissue radioactivity sample:normal (S:N) ratios than control (P less than 0.05), suggesting increased tissue injury. However, by several other measures of tissue damage, the two groups did not differ. MP-elevated PYP S:N ratios were explained by reduced PYP activity in normal myocardium of MP dogs. Further experiments in 21 dogs revealed that renal PYP clearance, which correlated with glomerular filtration rate (GFR) as measured by creatinine clearance, was increased in Mp dogs, resulting in accelerated urinary excretion of PYP (46.9+/-3.6 vs 35.8+/-2.4 percent injected dose in one hour, P less than 0.01), and reduced blood PYP. Thus MP does not modify countershock-induced myocardial injury. However, by increasing GFR, MP increased PYP excretion, resulting in lowered blood and normal zone myocardial PYP, thereby spuriously affecting myocardial PYP tissue uptake data.

Pancreatic uptake of 99Tc stannous pyrophosphate in experimental viral pancreatitis in mice.

The pancreatic uptake of 99mTc stannous pyrophosphate (99mTc-PYP) in acute pancreatitis in mice induced by coxsackievirus B3 was studied. 99mTc-PYP uptake ratio, measured by the ratio of counts/min/g for the pancreas to counts/min/g for the skull, began to increase 2 days after virus inoculation and markedly increased on day 3 when necrosis of pancreatic acinar cells was evident. On day 5, 99mTc-PYP ratio reached a maximum (2.95 +/- 1.85, mean +/- SD) and histologically fine granules of calcification were seen in the necrotic acinar cells. Thereafter, cellular infiltrations increased and were most severe on day 7, but 99mTc-PYP ratio had began to decrease. On day 28, the necrotic areas of the pancreas were replaced with fatty cells and calcification was still present, though 99mTc-PYP ratio had decreased. The present findings may provide a basis for 99mTc-PYP imaging in cases of clinical viral pancreatitis.

99mTc bone scanning agents--II. Adsorption of 99mTc(Sn)pyrophosphate complexes on the mineral phase of bone.

The adsorption of pyrophosphate, tin-pyrophosphate and 99mTc(Sn)pyrophosphate on Ca3(PO4)2 was investigated at pH 4.0 and pH 7.4. All components were radioactively labeled. Tin and reduced technetium were in most cases almost completely bound. The adsorption of pyrophosphate, tin(II) and technetium-99m at pH 4.0 was higher than at pH 7.4. The presence of tin gave rise to an increase of the pyrophosphate adsorption that was much larger than can be accounted for by a stoichiometric adsorption of tin-pyrophosphate. It is concluded that tin and technetium are bound as negatively charged complexes with pyrophosphate. Finally it is argued that the fraction of the bone scanning agent that reaches the bone surface is adsorbed completely by the mineral phase.

Technetium-99m stannous pyrophosphate imaging of experimental infective endocarditis.

Technetium-99m stannous pyrophosphate (99mTc-PYP) cardiac scintigraphy was performed in 15 rabbits with experimental Streptococcus sanguis aortic valve infective endocarditis. The animals were imaged five to seven days after the administration of bacteria, and in each case abnormal accumulation of the tracer was visualized in the region of the aortic valve. Three types of cardiac scintigraphic patterns were demonstrated: focal, multifocal and extensive, each correlating well with the anatomical extent of the lesion as defined by gross pathology. Tissue distribution studies demonstrated a 30 +/- 5.3 (mean +/- SEM) fold excess of radionuclide uptake in the infective endocarditis lesion compared with that of normal myocardium. Imaging of excised hearts from four animals showed an excellent correlation with in vivo imaging as well as gross pathology. Five animals with nonbacterial thrombotic aortic valve endocarditis demonstrated similar scintigraphic and tissue distribution results. In contrast, four normal animals failed to demonstrate abnormal 99mTc-PYP cardiac scintigrams or tissue uptake. This study demonstrates that 99mTc-PYP cardiac scintigraphy is a sensitive technique to detect experimental aortic valve endocarditis.

Acute myocardial infarction: clinical application of technetium 99m stannous pyrophosphate infarct scintigraphy.

Acute myocardial infarction is being recognized as a spectrum of clinical subsets. This appreciation has been brought about to a large degree by the development of several new tools that can be applied clinically to aid in evaluation of patients with acute infarction, and in some cases to provide short and long-term prognostic information. In the realm of noninvasive methods, several tests utilizing radiopharmaceuticals and scintillation cameras have emerged and are rapidly becoming reliable diagnostic parameters in patients with coronary disease and infarction. Technetium 99m (stannous) pyrophosphate (TcPYP) scintigraphy, one of the first of these techniques to find clinical use, has been shown to be an accurate indicator of acute transmural myocardial infarction and provides added sensitivity and specificity to the diagnosis. Increased diagnostic accuracy, the dimension of visible localization and the potential for infarct sizing promise physicians better understanding of a patient's clinical presentation and a more rational approach to management.