Isothiocyanate-trigalactose: application for antibody-targeted delivery of diagnostic and therapeutic agents.

Radiolabeled monoclonal antibodies (MAb) and MAb-streptavidin conjugates exhibit slow blood clearance which impedes radioimmunoimaging and radioimmunotherapy. To control blood clearance and lower background levels, lesion-specific targeting proteins can be modified with galactose derivatives for liver uptake via the hepatocyte galactose receptor. In this study, an isothiocyanate-trigalactose derivative (ITC-Tgal) designed for direct coupling to protein amino groups, was synthesized and characterized. In vitro experimentation demonstrated efficient conjugation of ITC-Tgal to streptavidin (SA) and MAb Fab fragment with a corresponding decrease in protein net charge. In vivo studies were conducted with radiolabeled ITC-Tgal modified and native SA and MAb Fab fragment. ITC-Tgal modified SA and Fab fragment exhibited increased blood clearance with the liver uptake and the rate of blood clearance controlled by the extent of ITC-Tgal modification.

Effect of angiotensin converting enzyme inhibition on the Doppler waveform in dogs with renal artery stenosis.

Our objective was to investigate whether the angiotensin converting enzyme inhibitor enalaprilat improves detection of hemodynamically significant renal artery stenoses in dogs. Renal artery stenoses of 50 to 99% were surgically created unilaterally in five dogs. Doppler ultrasonographic evaluation was performed at baseline (no stenosis), after creation of the stenosis, and after the administration of enalaprilat. The resistive index increased in the nonstenotic kidney (P < 0.01) but not in the stenotic kidney after administration of enalaprilat. The difference in resistive indices between nonstenotic and stenotic kidneys increased significantly (P < 0.05) after administration of enalaprilat. Measurement of the resistive index after administration of an angiotensin converting enzyme inhibitor in humans may improve the performance of Doppler ultrasonography in detecting hemodynamically significant renal artery stenoses.

Sonography: a useful tool to detect the mechanical causes of renal transplant dysfunction.

PURPOSE: The purpose of this study was to evaluate the utility of sonography in distinguishing between mechanical and nonmechanical causes for renal transplant dysfunction. METHODS: We reviewed all ultrasound examination reports (n = 286) for 63 consecutive patients who received 64 renal transplants. We assessed the sensitivity and specificity of different degrees of hydronephrosis (mild, moderate, or severe) in detecting urinary tract obstruction; different volumes of new or increasing peritransplant fluid in detecting urine leaks; different total volumes of peritransplant fluid in predicting significant compression of the transplant; and Doppler vascular criteria for predicting arterial and venous occlusion. RESULTS: All mechanical complications were detected (100% sensitivity) with specificities of 91.9% for ureteral obstruction (criterion, moderate hydronephrosis), 83.4% for urine leaks (criterion, any new fluid or any increase), 91.4% for fluid collections that compressed the transplant (criterion, > 100 ml), and 100% for vascular occlusion (criteria, no flow for arterial occlusion; no venous flow and reversal of arterial flow during diastole for venous occlusion). CONCLUSIONS: Sonography is very useful in distinguishing between mechanical and nonmechanical causes for renal transplant dysfunction. It has high sensitivity and acceptable specificity in this setting.

Magnetic resonance imaging of the hip with a pelvic phased-array surface coil: a technical note.

OBJECTIVE: The aim of this study was to assess the capability of high-resolution images obtained with a commercially available pelvic phased-array surface coil to demonstrate normal hip anatomy. DESIGN: We retrospectively analyzed the oblique coronal magnetic resonance (MR) images of hips of 36 consecutive patients acquired on a 1.5-T clinical imager using a pelvic phased-array coil as a receiver, a 16-20 cm field of view, and 5 mm slice thickness. PATIENTS: Thirty-six patients were studied, age 15-81 years. There were 20 males and 16 females. RESULTS AND CONCLUSIONS: The articular cartilage, cortex, superior labrum, and iliofemoral ligament were well visualized on proton density weighted fat saturation (PDF) images. The femoral and obturator vessels, obturator nerve, and various muscles were easily seen on T1-weighted images. High-resolution imaging of the hip is achievable in a reasonable amount of time using newer phased-array surface coils and may play an increasing role in the future evaluation of hip disorders.

Interobserver and intraobserver variation in determining intrarenal parameters by Doppler sonography.

OBJECTIVE: This study identified the extent of inter- and intraobserver variation in assessing intrarenal Doppler waveform parameters. SUBJECTS AND METHODS: Optimized Doppler waveforms from the interlobar arteries of five healthy volunteers were recorded. The waveforms were digitized for off-line evaluation by six observers. Using the same set of waveforms, the observers initially selected three of six waveforms for each subject on the basis of their preference for clinical usefulness. The observers then were directed to evaluate three previously picked waveforms. The systolic acceleration time (SAT) and the resistive index were calculated from each waveform. The observers also characterized the shape of each waveform according to four categories. RESULTS: Significant interobserver (p < .01) but not intraobserver variation occurred when measuring the SAT. Differences were caused by the observers' perceptions of waveform shapes (kappa = .12, p < .01) as well as the observers choosing different systolic starting points. No significant inter- or intraobserver variation occurred in measuring the resistive index. CONCLUSION: Much variability among and within laboratories can be expected when observers of Doppler waveforms use SAT to evaluate patients for renal artery stenosis.

Biochemical modification of streptavidin and avidin: in vitro and in vivo analysis.

UNLABELLED: The high affinity streptavidin (or avidin)/biotin system is being investigated for imaging and radiotherapy procedures. Streptavidin (SA) and avidin exhibit markedly different pharmacokinetics, with avidin clearing from the blood much faster than SA. To optimize blood clearance kinetics, SA and avidin were biochemically modified and analyzed in vitro and in vivo. METHODS: Galactose moieties were covalently attached to promote binding by hepatocyte galactose receptors and hasten SA clearance. To prolong avidin clearance, avidin was deglycosylated and/or neutralized by acetylation of its lysine amino acids. In vitro, the modified proteins were analyzed by isoelectric focusing, SDS polyacrylamide electrophoresis and a biotin binding saturation assay. The modified and native proteins were radiolabeled with 131I and injected into rabbits for pharmacokinetic, redistribution and imaging analysis. RESULTS: For SA, the resulting increase in blood clearance and liver accumulation was correlated to the amount of galactose bound to SA. For avidin, each type of modification increased its circulation time, with the slowest clearance resulting from a combination of deglycosylation and neutralization. CONCLUSION: Biochemical modification of SA and avidin resulted in altered pharmacokinetics compared to the native proteins. Modified SA or avidin, when cross-linked with a lesion-specific targeting agent, may be applicable for rapid two-step in vivo imaging techniques.

Quantification and lowering of serum biotin.

An enzyme-linked assay was developed to quantify serum biotin concentrations in experimental animals and humans. With this assay the effect on serum biotin concentration after intravenous injection of streptavidin or the addition of avidin to food was studied in rabbits and dogs. Intravenous injection of streptavidin reduced serum biotin values quickly but temporarily, in a dose-dependent manner in both species. Addition of avidin to the diet lowered biotin values approximately four- to fivefold in the rabbits, with resultant biotin concentrations similar to those of humans.

Effect of a main renal artery stenosis on the downstream Doppler waveform in dogs.

RATIONALE AND OBJECTIVES: We evaluated the changes in the down-stream Doppler waveforms caused by a proximal stenosis in the main renal artery of dogs. METHODS: Renal parenchymal arterial waveforms downstream from mild (< 50%), moderate (50-75%), and severe (76-95%) stenoses were compared with nonstenotic baseline waveforms in five mongrel dogs. Waveform shapes were categorized as biphasic or monophasic. The percentage of biphasic and monophasic waveforms was determined for each stenosis. The acceleration index (AI) and the acceleration time (AT) were determined using traditional and modified calculations (AI' and AT'). Late systolic deceleration (DS), diastolic deceleration (DD), and the resistive index (RI) also were measured. RESULTS: AT, AI', and AT' demonstrated significant differences between the severe stenoses and nonstenotic baselines (p < .05); however, there was no difference between the mild and moderate stenoses versus baselines. The percentage distribution of monophasic and biphasic waveforms was highly correlated with the degree of stenosis. Monophasic waveforms increased on average from 22.5% of baseline waveforms to 76.5% of waveforms in the severe stenoses. Biphasic waveforms decreased on average from 69.9% of baseline waveforms to 18.7% of waveforms in the severe stenoses. CONCLUSION: Quantitative evaluation of the downstream waveform parameters (AI, AT, AI', AT', DS, DD, and RD in the dog kidney is not sufficiently accurate, but calculation of the percentage of the monophasic and biphasic waveforms present may be useful to predict a hemodynamically significant renal artery stenosis (> or = 50%).

Fluence rate effects in photodynamic therapy of multicell tumor spheroids.

EMT6/Ro spheroids approximately 500 microns in diameter were subjected to photodynamic therapy administered at various incident radiation fluence rates. Following 24 h incubation with 10 micrograms/ml Photofrin, groups of spheroids were irradiated at 630 nm with an identical fluence of 60 J/cm2, delivered at fluence rates ranging from 25 to 200 mW/cm2. After treatment, spheroids were dissociated, cell yields were determined, and surviving cells were assayed for their colony-forming ability. A surviving fraction was calculated for each treatment group by computing the product of the fractional cell yield and the plating efficiency. The results exhibit a strong dependence on the fluence rate, with surviving fractions decreasing from approximately 0.5 to 0.07 as the incident fluence rate was lowered from 200 to 25 mW/cm2. These data were analyzed using a mathematical model of photochemical oxygen consumption in spheroids undergoing photodynamic therapy. Calculations showed that therapy-induced oxygen consumption creates hypoxic volumes within which cells would be protected from singlet oxygen-mediated damage and that the magnitude of these hypoxic volumes depends on the radiation fluence rate. The fluence rate dependence of the spheroid cell survival was consistent with such an interpretation.