Novel fluorescence-based approaches for the study of biogenic amine transporter localization, activity, and regulation.

Pre-synaptic norepinephrine (NE) and dopamine (DA) transporters (NET and DAT) terminate catecholamine synaptic transmission through reuptake of released neurotransmitter. Recent studies reveal that NET and DAT are tightly regulated by receptor and second messenger-linked signaling pathways. Common approaches for studying these transporters involve use of radiolabeled substrates or antagonists, methods possessing limited spatial resolution and that bear limited opportunities for repeated monitoring of living preparations. To circumvent these issues, we have explored two novel assay platforms that permit temporally resolved quantitation of transport activity and transporter protein localization. To monitor the binding and transport function of NET and DAT in real-time, we have investigated the uptake of the fluorescent organic compound 4-(4-diethylaminostyryl)-N-methylpyridinium iodide (ASP+). We have extended our previous single cell level application of this substrate to monitor transport activity via high-throughput assay platforms. Compared to radiotracer uptake methods, acquisition of ASP+ fluorescence is non-isotopic and allows for continuous, repeated transport measurements on both transfected and native preparations. Secondly, we have extended our application of small-molecule-conjugated fluorescent CdSe/ZnS nanocrystals, or quantum dots (Qdots), to utilize antibody and peptide ligands that can identify surface expressed transporters, receptors and other membrane proteins in living cell systems. Unlike typical organic fluorophores, Qdots are highly resistant to bleaching and can be conjugated to multiple ligands. They can also be illuminated by conventional light sources, yet produce narrow, gaussian emission spectra compatible with multiple target visualization (multiplexing). Together, these approaches offer novel opportunities to investigate changes in transporter function and distribution in real-time with superior spatial and temporal resolution.

Pyomyositis. Increasing recognition in temperate climates.

Pyomyositis is common in the tropics yet is rarely reported in temperate climates. A woman in whom pyomyositis developed in a temperate climate is presented. Computed tomography was the key in the diagnosis of the disease involving the muscles of the left lateral chest wall. The patient's condition responded to intravenous antibiotics and open abscess drainage. The 31 cases reported in the United States are reviewed. Unfamiliarity still poses a barrier to early diagnosis, although pyomyositis is being more frequently described in temperate climates.

A precision cranial immobilization system for conformal stereotactic fractionated radiation therapy.

PURPOSE: Conformal radiotherapy has been shown to benefit from precision alignment of patient target to therapy beam (1, 6, 13). This work describes an optimized immobilization system for the fractionated treatment of intracranial targets. A study of patient motion demonstrates the high degree of immobilization which is available. METHODS AND MATERIALS: A system using dental fixation and a thermoplastic mask that relocates on a rigid frame is described. The design permits scanning studies using computed tomography (CT) and magnetic resonance imaging (MR), conventional photon radiotherapy, and high precision stereotactic proton radiotherapy to be performed with minimal repositioning variation. Studies of both intratreatment motion and daily setup reliability are performed on patients under treatment for paranasal sinus carcinoma. Multiple radiographs taken during single treatments provide the basis for a three-dimensional (3D) motion analysis. Additionally, studies of orthogonal radiographs used to setup for proton treatments and verification port films from photon treatments are used to establish day to day patient position variation in routine use. RESULTS: Net 3D patient motion during any treatment is measured to be 0.9 +/- 0.4 mm [mean +/- standard deviation (SD)] and rotation about any body axis is 0.14 +/- 0.67 degrees (mean +/- SD). Day-to-day setup accuracy to laser marks is limited to 2.3 mm (mean) systematic error and 1.6 mm (mean) random error. CONCLUSION: We conclude that the most stringent immobilization requirements of 3D conformal radiotherapy adjacent to critical normal structures can be met with a high precision system such as the one described here. Without the use of pretreatment verification, additional developments in machine and couch design are needed to assure that patient repositioning accuracy is comparable to the best level of patient immobility achievable.

Efficacy of diltiazem for control of symptoms of coronary arterial spasm.

To evaluate the efficacy of the calcium antagonist diltiazem for therapy of active coronary arterial spasm, 13 patients with clinical variant angina attributed to documented coronary arterial spasm completed a prospective randomized double-blind crossover trial of diltiazem (120 and 240 mg/day) versus placebo. Response was assessed with the diary technique measuring frequency of angina, consumption of nitroglycerin and percent of pain-free days. When 120 mg of diltiazem/day was compared with the paired placebo period there was a significant increase in percent of pain-free days (from 43 to 71 percent [p = 0.03]), but no significant decrease in frequency of angina (p = 0.06) or consumption of nitroglycerin (p = 0.32). When 240 mg of diltiazem/day was compared with the paired placebo period there was a significant increase in percent of pain-free days (from 50 to 79 percent [p = 0.03]) and a significant decrease in both frequency of angina (from 1.6 to 0.4 episodes/day [p = 0.03]) and consumption of nitroglycerin (from 1.3 to 0.4/day [p = 0.01]). Diltiazem was found to be a highly effective drug for control of symptoms of active coronary arterial spasm, without side effects and with excellent patient tolerance.

Comparison of two detail screen-film systems using a rheumatoid erosion model.

Two detail screen-film systems, Lanex Fine/Ortho M and Lanex Fine/Ortho G, were evaluated using a radiographic model for rheumatoid erosions. Radiographs of hand bones with cortical defects using both screen-film systems were obtained. Seven observers analyzed 48 sites on each of 22 films for the presence of these cortical defects. Using analysis of variance techniques and receiver operating characteristic curves, no significant difference was found between the two systems for the detection of these subtle cortical defects. The authors conclude that although the Lanex Fine/Ortho M system provides better definition of laboratory test objects than Lanex Fine/Ortho G, the two systems do not differ when used for detection of cortical lesions.

Comparison of computed tomography and ultrasound in abdominal staging of renal cancer.

The relative accuracy of computed tomography (CT) and ultrasound in abdominal staging of renal cancer was determined in 22 patients. CT is capable of detecting tumor invasion of perinephric fat and adjacent muscles, which cannot usually be shown by ultrasound. While both CT and ultrasound demonstrate venous and retroperitoneal tumor extension, CT is more reliable since bowel gas not infrequently obscures the retroperitoneum on ultrasonic scanning. However, ultrasound will often provide valuable information; and whenever a solid renal mass is detected by echography using prone scans, abdominal scans should be obtained for staging pruposes.

Comparison of proton and x-ray conformal dose distributions for radiosurgery applications.

Highly focused dose distributions for radiosurgery applications are successfully achieved using either multiple static high-energy particle beams or multiple-arc circular x-ray beams from a linac. It has been suggested that conformal x-ray techniques using dynamically shaped beams with a moving radiation source would offer advantages compared to the use of only circular beams. It is also thought that, generally, charged particle beams such as protons offer dose deposition advantages compared to x-ray beams. A comparison of dose distributions was made between a small number of discrete proton beams, multiple-arc circular x-ray beams, and conformal x-ray techniques. Treatment planning of a selection of radiosurgery cases was done for these three techniques. Target volumes ranged from 1.0-25.0 cm3. Dose distributions and dose volume histograms of the target and surrounding normal brain were calculated. The advantages and limitations of each technique were primarily dependent upon the shape and size of the target volume. In general, proton dose distributions were superior to x-ray distributions; both shaped proton and shaped x-ray beams delivered dose distributions which were more conformal than x-ray techniques using circular beams; and the differences between all proton and x-ray distributions were negligible for the smallest target volumes, and greatest for the larger target volumes.

Vena caval occlusion after bird's nest filter placement.

BACKGROUND: Inferior vena caval thrombosis as a result of intracaval barrier devices occurs in 6.5% of patients with Greenfield filters. The incidence is less well defined in patients in whom bird's nest filters have been placed. We reviewed our experience with bird's nest filters to determine the incidence of filter-induced caval thrombosis. METHODS: The records of 140 patients with bird's nest filters were reviewed, living patients were interviewed, and the inferior vena cava examined in 37 patients by duplex scanning. RESULTS: Ninety-three patients were available for evaluation. Five of these patients were found to have caval thrombosis by duplex scanning and 2 had clinical symptoms and signs compatible with caval thrombosis. The majority of these patients were on anticoagulants at the time of filter thrombosis. CONCLUSION: The incidence of filter-induced vena caval thrombosis in patients with bird's nest filters (7%) is comparable with that of Greenfield filters. Because of the catastrophic manifestations of this complication and the increasing application of vena caval filters, the role of these filters in the treatment of thrombotic disease needs further critical evaluation.

The measurement of proton stopping power using proton-cone-beam computed tomography.

A cone-beam computed tomography (CT) system utilizing a proton beam has been developed and tested. The cone beam is produced by scattering a 160 MeV proton beam with a modifier that results in a signal in the detector system, which decreases monotonically with depth in the medium. The detector system consists of a Gd2O2S:Tb intensifying screen viewed by a cooled CCD camera. The Feldkamp-Davis-Kress cone-beam reconstruction algorithm is applied to the projection data to obtain the CT voxel data representing proton stopping power. The system described is capable of reconstructing data over a 16 x 16 x 16 cm3 volume into 512 x 512 x 512 voxels. A spatial and contrast resolution phantom was scanned to determine the performance of the system. Spatial resolution is significantly degraded by multiple Coulomb scattering effects. Comparison of the reconstructed proton CT values with x-ray CT derived proton stopping powers shows that there may be some advantage to obtaining stopping powers directly with proton CT. The system described suggests a possible practical method of obtaining this measurement in vivo.

Grey-scale ultrasonic characteristics of carcinoma of the pancreas.

Fifty-five sonograms were performed on 25 cases of surgically and histologically proven carcinoma of the pancreas. The sonographic appearances of the tumour were determined. A diffuse echogenic pattern with irregular margin was the most common appearance. Displacement and indentation of the inferior vena cava and superior mesenteric vessels, greatly assist in localizing the tumour to the pancreas. A reliable prediction of tumour size and resectability cannot be determined by sonography alone. Sonography is helpful in assessing the response of unresectable pancreatic carcinoma to therapy. A more sonolucent appearance of the tumour occurs following chemotherapy.

Photon-enhanced thermionic emission from heterostructures with low interface recombination.

Photon-enhanced thermionic emission is a method of solar-energy conversion that promises to combine photon and thermal processes into a single mechanism, overcoming fundamental limits on the efficiency of photovoltaic cells. Photon-enhanced thermionic emission relies on vacuum emission of photoexcited electrons that are in thermal equilibrium with a semiconductor lattice, avoiding challenging non-equilibrium requirements and exotic material properties. However, although previous work demonstrated the photon-enhanced thermionic emission effect, efficiency has until now remained very low. Here we describe electron-emission measurements on a GaAs/AlGaAs heterostructure that introduces an internal interface, decoupling the basic physics of photon-enhanced thermionic emission from the vacuum emission process. Quantum efficiencies are dramatically higher than in previous experiments because of low interface recombination and are projected to increase another order of magnitude with more stable, low work-function coatings. The results highlight the effectiveness of the photon-enhanced thermionic emission process and demonstrate that efficient photon-enhanced thermionic emission is achievable, a key step towards realistic photon-enhanced thermionic emission based energy conversion.

Differential serotonin transport is linked to the rh5-HTTLPR in peripheral blood cells.

The human serotonin transporter (SERT) gene possesses a 43-base pair (bp) insertion-deletion promoter polymorphism, the h5-HTTLPR. Genotype at this locus correlates with variation in anxiety-related personality traits and risk for major depressive disorder in many studies. Yet, the complex effects of the h5-HTTLPR, in combination with closely associated single-nucleotide polymorphisms (SNPs), continue to be debated. Moreover, although SERT is of high clinical significance, transporter function in vivo remains difficult to assess. Rhesus express a promoter polymorphism related to the h5-HTTLPR. The rh5-HTTLPR has been linked to differences in stress-related behavior and cognitive flexibility, although allelic variations in serotonin uptake have not been investigated. We studied the serotonin system as it relates to the 5-HTTLPR in rhesus peripheral blood cells. Sequencing of the rh5-HTTLPR revealed a 23-bp insertion, which is somewhat longer than originally reported. Consistent with previous reports, no SNPs in the rh5-HTTLPR and surrounding genomic regions were detected in the individuals studied. Reductions in serotonin uptake rates, cell surface SERT binding, and 5-hydroxyindoleacetic acid/serotonin ratios, but not SERT mRNA levels, were associated with the rh5-HTTLPR short allele. Thus, serotonin uptake rates are differentiable with respect to the 5-HTTLPR in an easily accessible native peripheral tissue. In light of these findings, we foresee that primary blood cells, in combination with high sensitivity functional measurements enabled by chronoamperometry, will be important for investigating alterations in serotonin uptake associated with genetic variability and antidepressant responsiveness in humans.

Gray scale ultrasonic appearances of renal transplant rejection.

Ultrasonic examination of renal transplants with special attention to the parenchymal echo pattern has been undertaken in 43 patients. In the normal renal transplant, the pyramids, cortex and renal sinus can be distinguished. Renal transplant rejection is manifested by swelling and decreased echogenicity of the pyramids and hyperechogenic cortex. In addition, large anechoic areas due to hemorrhagic infarcts and necrosis are seen. In long-standing rejection, a normal or small-sized kidney with an irregular intrarenal echo pattern is observed. In 13 cases of acute tubular necrosis, none of the above appearances could be demonstrated. Serial ultrasonic scans are essential to reveal evolutionary changes of the rejection process.

Use of mass spectrometer to measure CO2 and O2 fluxes in voltage-clamped epithelia.

A quadrupole mass spectrometer was coupled to an Ussing chamber in order to evaluate rates of oxidative metabolism in voltage-clamped epithelia. Well-defined mixing characteristics of the continuously perfused chamber allowed CO2 and O2 concentrations to be related to rates of CO2 efflux, JCO2, and oxygen influx, JO2. The use of a model tissue to simulate step changes in JCO2 validated the treatment, with response within a minute. Monitoring of metabolism was facilitated by use of a desk-top computer, which evaluated JCO2 at 6-s intervals. Concurrent measurements of electrical current and JCO2 were made in the toad urinary bladder in order to relate active sodium transport to metabolism; the use of amiloride to eliminate active transport and the associated metabolism then allowed evaluation of the rates of active Na transport (JNa) and suprabasal metabolism (JsbCO2), and their ratio JNa/JsbCO2. We report the ability to resolve a 5 pmol/s change in CO2 efflux or an 11 pmol/s change in O2 influx rates.

Time course of active Na transport and oxidative metabolism following transepithelial potential perturbation in toad urinary bladder.

The use of an Ussing chamber with well-defined mixing characteristics coupled to a mass spectrometer permits the concurrent evaluation of transepithelial current and oxidative metabolism with improved temporal resolution. The time-course of the amiloride-sensitive current Ia and the rate of suprabasal CO2 production JsbCO2 were observed in 10 toad urinary bladders at short-circuit and after clamping delta psi at 100 mV, serosa positive. Following perturbation of delta psi (0 leads to 100 mV), Ia declined sharply within 1/2 min, remaining near constant approximately 15 min, and then increased slightly. JsbCO2 declined more gradually, remained near constant at approximately 4-7 min, and then declined further. Detailed analysis revealed an early quasi-steady state with near constancy of JsbCO2 starting at 2.9 +/- 1.1 (SD) min and lasting 4.7 +/- 1.8 (SD) min, followed by relaxation to a later steady state at about 15 min. During the early quasi-steady state, Ia was also nearly constant. Considering that in steady states Ia/F approximately or equal to JaNa, the rate of transepithelial active Na transport, during the early quasi-steady state mean values +/- SE of JaNa, JsbCO2 and (JaNa/JsbCO2) were, respectively, 29.9 +/- 1.7%, 59.4 +/- 3.2%, and 56.4 +/- 5.7% of values at short-circuit. Corresponding values during the late steady state were 41.4 +/- 6.0%, 38.2 +/- 6.1%, and 111.3 +/- 8.6%. Thus the flow ratio JaNa/JsbCO2 was depressed significantly during the early quasi-steady state, but returned later to the original value. The results of measurements of Ia and JsbO2 in three hemibladders were qualitatively similar. In terms of a phenomenological "black-box" treatment the findings are consistent with earlier studies indicating incomplete coupling between transport and metabolism. Further studies will be required to clarify the molecular basis for these observations.

Phase inversion tissue harmonic sonographic imaging: a clinical utility study.

OBJECTIVE: We evaluated the utility of phase inversion tissue harmonic sonographic imaging in a variety of clinical applications. SUBJECTS AND METHODS: Two hundred randomly selected patients were scanned with both fundamental and tissue harmonic methods on a sonography system. The following transducers were used: 2.5 MHz, 20 mm phased array; 3 MHz, 40 mm curved array; 6.5 MHz, 13 mm curved array transvaginal; and 7.5 MHz, 40 mm linear. Operators evaluated visualization of normal and pathologic tissues by tissue harmonic versus fundamental imaging using scores ranging from 1 for much worse visualization to 5 for much better visualization. They also assessed the overall utility of tissue harmonic imaging in the diagnosis of the patient's condition. The studies were saved on magnetooptical disc and were independently reviewed by one of the authors. RESULTS: Tissue harmonic imaging was helpful for all types of examination. Tissue harmonic imaging improved visualization of normal tissue in 49% of the cases and pathologic tissue in 73% of the cases. Tissue harmonic imaging was found to be diagnostically helpful in 43% of the cases and essential to the diagnosis in 6% of the cases. CONCLUSION: Tissue harmonic imaging significantly improves visualization of both normal and pathologic tissues and its selective use has major diagnostic utility in a wide variety of clinical applications.

Exencephaly: sonographic findings and radiologic-pathologic correlation.

Exencephaly is a rare precursor of anencephaly in which a large amount of brain tissue is present despite the absence of the calvaria. It was discovered together with hydramnios in a near-term fetus. Sonographic, angiographic, and pathologic findings are discussed.