Identifying ureteropelvic junction obstruction by fluorescence imaging: a comparative study of imaging modalities to assess renal function and degree of obstruction in a mouse model.

PURPOSE: Radiological imaging is the mainstay of diagnosing ureteropelvic junction obstruction. Current established radiological modalities can potentially differentiate the varying degrees of obstruction but they are limited in functionality, applicability and/or comprehensiveness. Of particular concern is that some tests require radiation, which has long-term consequences, especially in children. MATERIALS AND METHODS: We investigated the novel use of Genhance™ 680 dynamic fluorescence imaging to assess ureteropelvic junction obstruction in 20 mice that underwent partial or complete unilateral ureteral obstruction. Ultrasound, mercaptoacetyltriglycine renography, magnetic resonance imaging and fluorescence imaging were performed. RESULTS: Our model of partial and complete obstruction could be distinguished by ultrasound, mercaptoacetyltriglycine renography and magnetic resonance imaging, and was confirmed by histological analysis. Using fluorescence imaging distinct vascular and urinary parameters were identified in the partial and complete obstruction groups compared to controls. CONCLUSIONS: Fluorescence imaging is a feasible alternative radiological imaging modality to diagnose ureteropelvic junction obstruction. It provides continuous, detailed imaging without the risk of radiation exposure.

Development of an ovine model of pediatric complete heart block.

BACKGROUND: Complete heart block is a significant clinical problem that can limit the quality of life in affected children. To understand the pathophysiology of this condition and provide for development of novel therapies, we sought to establish a large animal model of permanent, pacemaker-dependent atrioventricular block (AVB) that mimics the size and growth characteristics of pediatric patients. MATERIALS AND METHODS: We utilized nine immature lambs weighing 10.5 ± 1.4 kg. After implantation of dual-chamber pacemaker devices with fixed leads, AVB was produced by interrupting His-bundle conduction using radio-frequency ablation at the base of the non-coronary cusp of the aortic valve. Ablations (30 to 60 s in duration) were performed under fluoroscopic guidance with electrophysiological monitoring. Interrogation of pacemakers and electrocardiography (ECG) determined the persistence of heart block. Ovine hearts were also examined immunohistochemically for localization of conduction tissue. RESULTS: AVB was produced in eight animals using an atypical approach from the left side of the heart. One animal died due to ventricular fibrillation during ablation proximal to the tricuspid annulus and one lamb was sacrificed postoperatively due to stroke. Four sheep were kept for long-term follow-up (109.8 ± 32.9 d) and required continuous ventricular pacing attributable to lasting AVB, despite significant increases in body weight and size. CONCLUSIONS: We have created a large animal model of pediatric complete heart block that is stable and technically practicable. We anticipate that this lamb model will allow for advancement of cell-based and other innovative treatments to repair complete heart block in children.

Ultrasound diagnosis of mouse pregnancy and gestational staging.

Phenotypic analysis of mutant mice is limited by lack of accurate, simple, and nondestructive in utero imaging techniques. This study evaluated the usefulness of ultrasound imaging (US) to stage fetal mouse gestational age (GA) and depict morphologic development. We imaged 16 pregnant CD-1 mice and a total of 92 fetuses with a 15-MHz US transducer from 9.5 d postcoitus (DPC) until 20.5 DPC or delivery. Parameters recorded included gestational sac dimensions, crown-rump length (CRL), biparietal diameter (BPD), thoracoabdominal diameter (TAD), onset of cardiac activity, and morphologic development. At 9.5 d DPC, all gestations appeared as rounded sacs, with a diameter (mean +/- standard error) of 4.4 +/- 1 mm. BPD, CRL, and GA were highly correlated. The following structures were first identifiable at the following GA: cardiac activity, 10.5 DPC; major cardiovascular structures, 11.5 DPC; limb buds, 10.5 DPC; spine, 12.5 DPC; face and skull ossification, 13.5 DPC; rib ossification, 15.5 DPC; hind- and forelimb digits, 15.5 DPC; stomach and urinary bladder, 17.5 DPC; visualization of the rhombencephalic vesicle, 13.5 DPC; and visualization of the lateral ventricles, 14.5 DPC. The echogenic lungs were distinct from the liver as early as 12.5 DPC. The circle of Willis was detectable with color Doppler as early as 13.5 DPC and was easily visualized at 15.5 DPC. We found that US provides accurate, simple staging criteria for fetal mouse gestational development after 9.5 DPC and may be a nondestructive means of documenting phenotypic alterations in mutant mice in utero.

Fetal sheep development on ultrasound and magnetic resonance imaging: a standard for the in utero assessment of models of congenital abnormalities.

OBJECTIVE: To establish normative data for the size, conspicuity, and imaging characteristics of normal developing fetal sheep organs on ultrasound (US) and magnetic resonance (MR) imaging. METHODS: US and MR images of ten normal pregnant sheep, at 40, 65, 90, 115, and 140 gestational days (term = 145 days), were scored for organ conspicuity and imaging characteristics. Imaging biometry was correlated with specimens. Gestational age-based growth parameters were modeled using regression. RESULTS: Imaging biometry showed excellent correlation with specimens. Kidney, bladder, stomach, lung, liver, and spine were seen well from 65 days to term by US. More organs were consistently visible from 90 days to term by MR than by US. Most organ imaging characteristics tended not to change throughout gestation. CONCLUSION: Normal fetal sheep biometry, organ conspicuity, and imaging characteristics are established for US and MR and have potential use for the in utero assessment of sheep models of congenital abnormalities.

Radiation exposure reduction during voiding cystourethrography in a pediatric porcine model of vesicoureteral reflux.

PURPOSE: To compare grid-controlled variable-rate pulsed fluoroscopy (GCPFL) and continuous fluoroscopy (CFL) for the reduction of radiation exposure during voiding cystourethrography (VCUG) in a pediatric porcine model of vesicoureteral reflux. MATERIALS AND METHODS: Institutional animal care and use committee approval was obtained. Vesicoureteral reflux was simulated in four pigs, and 48 VCUG studies were performed (24 with GCPFL, 24 with CFL). VCUG was performed at abdominal girths of 8-10 cm (group 1, simulates human newborn to 6-month-old infant), 12-13 cm (group 2, simulates 2-3-year-old child), and 15-17 cm (group 3, simulates 10-year-old child). An electronic device calculated total radiation exposure during fluoroscopy and image recording. With five-point ordinal scales, VCUG images were scored independently for anatomic conspicuity and overall diagnostic quality by two radiologists (radiologists A and B). An analysis of variance was used to compare radiation exposures and fluoroscopy times between GCPFL and CFL and to determine whether radiation exposure and fluoroscopy time were dependent on the pig's abdominal girth. The Pearson product-moment correlation coefficient was used to assess whether fluoroscopy time was correlated with radiation exposure. Anatomic conspicuity and diagnostic quality scores were compared by means of the Wilcoxon signed rank test. RESULTS: Results of analysis of variance revealed that GCPFL resulted in a significant reduction in total radiation exposure compared with CFL for each of the three groups (P < .05 for each comparison), and this reduction was most marked in the larger animals. There were no significant differences in diagnostic quality of the recorded VCUG images (P > .05). Anatomic conspicuity was not significantly different for groups 2 and 3, but there was a significantly higher score for GCPFL in group 1 for radiologist A (P = .04). CONCLUSION: By using GCPFL in the performance of VCUG in a pediatric porcine model of vesicoureteral reflux, total radiation exposure can be reduced by a factor of 4.6-7.5 lower than with CFL, and diagnostic-quality images can be obtained.

Epinephrine prevents muscle blood flow increases after perineural injection of tetrodotoxin.

BACKGROUND: The local anesthetic properties of tetrodotoxin, a potent naturally occurring sodium channel blocker, have been recently reexamined. It was found that sciatic nerve block duration could be greatly increased and systemic toxicity greatly decreased if epinephrine was injected with tetrodotoxin. The mechanism that underlies epinephrine-mediated prolongation of tetrodotoxin nerve blocks is not known, but indirect evidence suggests that epinephrine may slow the clearance of tetrodotoxin from the site of injection. The authors hypothesized that tetrodotoxin causes vasodilatation at its injection site, which accelerates its systemic uptake, and that this vasodilatation is attenuated by coinjected epinephrine. METHODS: The radiolabeled microsphere technique was used to measure tissue blood flow in anesthetized rats before and after perisciatic injection of tetrodotoxin alone and in combination with epinephrine. RESULTS: Tetrodotoxin, in a dose of 0.1 ml of a 60 microM solution, significantly increased blood flow in perisciatic muscle at the injected side compared with simultaneous contralateral control and ipsilateral preinjection baseline. Tetrodotoxin did not increase blood flow in the sciatic nerve. Coinjection of epinephrine with tetrodotoxin prevented tetrodotoxin-induced increases in perisciatic muscle blood flow over time and did not alter sciatic nerve blood flow. Arterial blood pressure was maintained with this dose of tetrodotoxin, although brain blood flow decreased. Coinjection of epinephrine with tetrodotoxin prevented decreases in brain blood flow. Higher doses of tetrodotoxin produced hypotension. CONCLUSIONS: Vasoconstriction in the perisciatic muscles by epinephrine may contribute to the prolongation of tetrodotoxin-induced sciatic nerve blocks and the reduction of systemic toxicity of tetrodotoxin.

Normal and ischemic epiphysis of the femur: diffusion MR imaging study in piglets.

PURPOSE: To evaluate normal diffusion characteristics in the femur in piglets and changes in diffusion with increasing duration of femoral head ischemia. MATERIALS AND METHODS: Normal epiphyses, physes, and metaphyses of piglets were evaluated with line-scan diffusion imaging (n = 12) and diffusion-tensor imaging (n = 4). Apparent diffusion coefficient (ADC) differences between normal proximal and distal femoral structures, epiphyseal and physeal cartilage, and epiphyseal and metaphyseal marrow were compared (Mann-Whitney test). Short-term femoral ischemia was investigated after maximal abduction of the hips for 3 hours (n = 6); ADCs before and after abduction were compared (Wilcoxon signed rank test). Prolonged ischemia was investigated with placement of a ligature around the neck of a femur (n = 7); the ADC of the femur in this condition was compared (Wilcoxon signed rank test) with that of the normal contralateral femur. Changes in ADC ratios at three durations of ischemia (Kruskal-Wallis test) were compared. RESULTS: ADC was greater in epiphyseal cartilage (mean +/- 1 SD, 1.62 x 10(-3) mm2/sec +/- 0.38) than it was in physeal cartilage (1.28 x 10(-3) mm2/sec +/- 0.31) (P <.007) and greater in epiphyseal marrow (1.26 x 10(-3) mm2/sec +/- 0.38) than it was in metaphyseal marrow (0.91 x 10(-3) mm2/sec +/- 0.35) (P <.001). There was columnar arrangement of tensors in the physis. ADC decreased 26% after 3 hours of maximal abduction. After femoral neck ligature, ADC increased a mean of 27% after 6 hours and a mean of 75% after 96 hours. CONCLUSION: Normal line-scan diffusion imaging findings indicate relative restriction of diffusion in the metaphysis and parallel orientation of tensors in the physis. Diffusion is initially restricted with decreased blood flow but increases if ischemia lasts longer.

Correlation of contrast-enhanced power Doppler sonography and conventional angiography of abduction-induced hip ischemia in piglets.

OBJECTIVE: The purpose of this study was to determine if enhanced power Doppler sonography can detect early ischemia of the capital femoral epiphysis induced by hip hyperabduction in piglets and to correlate these findings with angiography. MATERIALS AND METHODS: Proximal femoral perfusion was evaluated in 18 studies of 10 piglet hips with unenhanced power Doppler sonography, enhanced power Doppler sonography with IV contrast agent, and digital angiography, in neutral position, hyperabduction, and after release to neutral position. Enhancement ratios between pixel intensities of power Doppler sonography and enhanced power Doppler images in each position were calculated. Angiograms were analyzed for differences in flow with changes in hip position. RESULTS: With the piglet in neutral position, power Doppler sonography revealed few vessels in the femoral head. Contrast administration resulted in a temporary marked increase in the visualization of vessels in the femoral head. Quantitative enhanced power Doppler sonography revealed a marked decrease in pixel intensity with abduction (p < 0.001) that was not apparent on unenhanced studies (p = 0.28). The enhancement ratio decreased from 0.45 (mean +/- SD, +/- 0.26) in neutral position to 0.10 (+/- 0.21) after abduction; it returned to 0.41 (+/- 0.14) after release of abduction (p < 0.001 for each comparison). Angiographic studies in hyperabduction revealed a variable level of ischemia. CONCLUSION: Enhanced power Doppler sonography can be used to visualize the vascular supply to the cartilaginous femoral head in piglets and can detect reversible ischemia induced by hip hyperabduction. These differences correlate with digital angiographic evidence of ischemia.

Activation of peripheral excitatory amino acid receptors decreases the duration of local anesthesia.

BACKGROUND: Postsurgical wound infiltration with the -methyl-d-aspartate receptor antagonist ketamine and bupivacaine can significantly prolong the duration of local anesthesia. One possible mechanism for this effect is that increased glutamate concentrations, caused by tissue damage, sensitize nociceptive primary afferent fibers through activation of peripheral excitatory amino acid receptors. METHODS: The effect of intramuscular injection of hyper-tonic glutamate (1,000 mm), dextrose (1,400 mm), glutamate (1,000 mm) with the broad spectrum excitatory amino acid receptor antagonist kynurenate (100 mm), or isotonic saline (155 mm) on the duration of masseter muscle afferent fiber blockade after lidocaine (37 mm [1%], 10 microl) infiltration, on muscle edema formation and on muscle blood flow was examined. RESULTS: Injection of either glutamate or dextrose significantly shortened the duration of lidocaine blocks compared with isotonic saline; however, block duration was significantly shorter after glutamate than after dextrose. Injection of glutamate, but not isotonic saline, dextrose, or glutamate with kynurenate, significantly decreased the mechanical threshold of muscle afferent fibers. Injection of glutamate, dextrose, or glutamate with kynurenate produced equivalent large, long-lasting (> 60 min) edemas with high initial peak extracellular water content. Peak extracellular water decreased more rapidly when kynurenate was coinjected with glutamate. Both glutamate and dextrose significantly increased muscle blood flow for 30 min after injection. Glutamate-induced increases in blood flow were attenuated by kynurenate. CONCLUSIONS: These results suggest that shortened lidocaine block durations observed after glutamate injection into the masseter muscle result from sensitization of afferent fibers as well as increases of peak extracellular water content and blood flow in masseter muscle. These effects of glutamate are mediated in part through activation of peripheral excitatory amino acid receptors.

The effects of lidocaine and adrenergic agonists on rat sciatic nerve and skeletal muscle blood flow in vivo.

UNLABELLED: It has been proposed that epinephrine prolongs lidocaine nerve blockade duration by exerting a local vasoconstrictive effect on tissues at the injection site, slowing lidocaine's local clearance. However, previous studies have failed to demonstrate consistent effects of lidocaine and epinephrine, injected alone and in combination, on vascular tone or regional blood flow. To reinvestigate this idea, in this study we used the radiolabeled microsphere technique to measure in vivo tissue blood flow before and at several time points after perisciatic nerve and intramasseter muscle injection of lidocaine alone, epinephrine, the selective alpha(1)-adrenergic receptor agonist phenylephrine, or lidocaine combined with these adrenergic receptor agonists. Repeated-measures analyses of variance were used to assess significant changes in blood flow over time. Lidocaine (2, 10, and 20 mg/mL) and epinephrine (10 micro g/mL or 1:100,000) injected alone did not alter blood flow in sciatic nerve, perisciatic muscle, or masseter muscle. Injections of lidocaine (10 mg/mL) combined with epinephrine (10 micro g/mL) did not affect adjacent muscle blood flow but caused a mild reduction in sciatic nerve blood flow, which was significant 30 min after injection. However, phenylephrine (10 micro g/mL), a potent vasoconstrictor, combined with lidocaine (10 mg/mL) significantly reduced blood flow in all three tissues. Our findings suggest that mechanisms other than local vasoconstriction may contribute to the prolongation of lidocaine nerve blocks by epinephrine. IMPLICATIONS: Accepting that the microsphere technique may be limited in its sensitivity to detect small but clinically relevant changes in tissue blood flow, our findings suggest that mechanisms other than local vasoconstriction may contribute to the prolongation of lidocaine nerve blocks by epinephrine.