Slow calcium and potassium currents in frog skeletal muscle: their relationship and pharmacologic properties

Slow Ca and K currents across frog skeletal muscle membrane were recorded with the Vaseline gap voltage clamp in order to investigate block by divalent cations and various organic compounds. Cd²⁺, Ni²⁺, Co²⁺, Mn²⁺, Mg²⁺ all block Ca currents, as do barbiturates, D-600 and nifedipine. Local anesthetics also block Ca currents, with the impermeant quaternary lidocaine derivative, QX-314, being more than an order of magnitude less potent than its permeant parent compound. Surprisingly, all agents that blocked Ca currents also blocked the slow K currents. To explain this pharmacologic parallel, one could suggest that K current is activated by Ca²⁺ appearing in the myoplasm due to the combination of Ca current and release from internal stores. While possibly correct for intact fibres, this hypothesis appears not to apply in our case where the myoplasm contained the Ca chelator EGTA at high concentration. Instead, K currents seem to be activated by a decrease in external [Ca²⁺]. In the transverse tubules, Ca current is known to cause [Ca²⁺] to decline to submicromolar concentrations, and evidence is presented that K currents are activated by Ca depletion from a restricted extracellular space. It is suggested that K currents flow through Ca channels that have become capable of passing monovalent cations after the tubules have become depleted of Ca²⁺.     

Effects of defibrillator implantation testing on myocardial metabolism.

BACKGROUND: The effects of fibrillation/defibrillation episodes (FDEs) during defibrillator implantation on myocardial metabolism were investigated at various defibrillation energies in patients with different cardiac pathologies. METHODS: Myocardial lactate extraction (MLE) was examined during defibrillation threshold (DFT) testing in patients with either coronary artery disease (CAD, n = 20) or non-ischemic cardiomyopathy (CM, n = 10). Defibrillation pulses were released 15 seconds after induced fibrillation. A test cycle of four FDEs separated by 2-minute intervals was applied in each case. RESULTS: Mean MLE decreased significantly from 28 +/- 4% before FDEs to 8 +/- 5% immediately after all episodes in CAD patients, but recovered to 27 +/- 7% within 2 minutes even in patients with reduced left-ventricular function. In patients with CM mean MLE decreased markedly from 29 +/- 3% to -11 +/- 3% immediately after each FDE but increased to baseline (33 +/- 8%) within the recovery period. MLE changes were independent of defibrillation energy in all cases. CONCLUSIONS: Myocardial lactate production, suggesting cardiac ischemia, was observed in patients with CM, but not in patients with CAD. But recovery of myocardial lactate extraction was not faster in CAD patients, indicating that the fixed FDE cycle used was well tolerated by all patients.     

Intrathecal Therapy for Cancer and Nonmalignant Pain: Patient Selection and Patient Management

Intrathecal drug delivery improves pain relief, reduces suffering, and enhances quality of life in the small proportion of patients who do not respond well to oral analgesics, including oral morphine. Although morphine is the “gold standard,” and the only drug approved for intrathecal pain therapy in the United States, off-label use of alternative agents appears promising, particularly in patients with neuropathic pain. Careful patient selection and management are significant determinants of successful treatment outcomes. Patient selection criteria for cancer and nonmalignant pain are similar; however, a more comprehensive psychological and social assessment is required for patients with nonmalignant pain. In addition, all patients (those with cancer or nonmalignant pain) must exhibit a positive response to an epidural or intrathecal screening test. A multidisciplinary team approach, involving psychologists, nurses, physical therapists, social workers, and spiritual leaders should be used to manage patients. Current practices for patient selection and management, screening tests, and dosing guidelines for intrathecal drug delivery systems are discussed.     

Evaluation of a new monocortical screw for anterior cervical fusion and plating by a combined biomechanical and clinical study

The purpose of this combined study was to evaluate the stability and safety of a new monocortical screw-plate system for anterior cervical fusion and plating (ACFP) according to Caspar in comparison with classical bicortical fixation. In the biomechanical part of the study two groups, each comprising six fresh human cadaveric spines (C4–C7), matched for bone mineral density, additionally resulting in almost the same mean age, were used. Range of motion and neutral zone were analyzed in flexion-extension, rotation (left, right) and lateral bending (left, right) using pure moments of ± 2.5 Nm for each specimen in the intact state, after discectomy at C5/6 and after discectomy at C 5/6 followed by bone grafting plus plating (Caspar plates), with either monocortical or bicortical screws. For all three motion planes, no significant difference could be found between the new monocortical and the bicortical fixation techniques. The clinical part of the study was performed as a prospective study on 30 patients suffering from symptomatic degenerative cervical disc disease in one segment. At the latest follow-up, no hardware- or graft-related complications were seen in any of the patients. Following these findings monocortical screw fixation can be recommended for the majority of anterior cervical fusion and plating procedures in degenerative disease, making the procedure quicker, easier, and safer. Bicortical screw fixation still has specific indications for multilevel stabilization, poor bone quality (osteoporosis, rheumatoid disease – as bicortical oversized rescue screw), unstable spines (trauma, tumour) and in particular for the realignment of kyphotic deformities (restoration of the normal lordotic curve). Due to the design of the study the results apply only to surgical treatment of monosegmental degenerative disc disease at the time. Received: 28 November 1998 Revised: 19 March 1999 Accepted: 29 May 1999     

A novel PET tracer for the imaging of αvβ3 and αvβ5 integrins in experimental breast cancer bone metastases

The aim of this study was the evaluation of ⁶⁸Ga‐DOTA‐E‐[c(RGDfK)]₂ as a novel PET tracer to image αvβ3 and αvβ5 integrins. For this purpose, DOTA‐E‐[c(RGDfK)]₂ was labeled with ⁶⁸Ga, which was obtained from a ⁶⁸Ge/⁶⁸Ga generator, purified by solid‐phase extraction and the radiochemical purity analyzed by radio‐RP‐HPLC. ⁶⁸Ga‐DOTA‐E‐[c(RGDfK)]₂ was obtained reproducibly in radiochemical yields of 60 ± 6% and with an excellent radiochemical purity of >99%. In nude rats bearing bone metastases after injection of MDA‐MB‐231 human breast cancer cells, biodistribution studies were performed to evaluate the accumulation of the radiotracer in selected organs, blood and bone metastases 0.5, 1, 2 and 3 h post injection. A rapid uptake into the bone metastases and rapid blood clearance was observed, resulting in tumor–blood ratios of up to 26.6 (3 h post injection) and tumor–muscle ratios of up to 7.9 (3 h post injection). A blocking experiment with coinjected αvβ3/αvβ5 antagonist showed the tumor uptake to be receptor‐specific. In an initial in vivo micro PET evaluation of the tracer using the same animal model, the bone metastasis was clearly visualized. These results suggest that ⁶⁸Ga‐DOTA‐E‐[c(RGDfK)]₂ is a promising PET tracer suitable for the imaging of αvβ3 and αvβ5 integrins in bone metastases. This novel PET tracer should be further evaluated concerning its usefulness for early detection of bone metastases and monitoring treatment response of these lesions. Copyright © 2011 John Wiley & Sons, Ltd.     

The modulation of intermanual interactions during the specification of the directions of bimanual movements

In two experiments bimanual movements with various combinations of target directions were studied by means of the timed-response procedure. The findings revealed an adaptive modulation of intermanual interactions during direction specifications depending on particular target directions. For symmetric movements intermanual correlations of movement directions are positive, indicating a symmetric coupling. For parallel movements the positive intermanual correlations, observed at short preparation intervals, turn into negative correlations as the time available for motor preparation increases. Biases of mean directions, that can be observed for movements to targets with different eccentricities, reflect one or the other kind of coupling, symmetrical for symmetric target directions and parallel for parallel target directions. These biases are static, that is, they are present at long preparation times, and they are phasically enhanced at shorter preparation intervals. The task-adaptive modulation of intermanual interactions is superposed on a basic symmetry bias.

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In vivo Gd-DTPA concentration for MR lung perfusion measurements: Assessment with computed tomography in a porcine model

A linear relationship between MR signal and contrast-agent concentration (CAC) of the arterial-input function (AIF) is crucial for MR lung-perfusion quantification. The aim was to determine the in-vivo real maximum CAC of the AIF, using cine CT measurements in a porcine model. A dilution series (Gd-DTPA, 0-20 mM) was examined by clinical time-resolved 3D-GRE MRI and by MDCT in cine CT mode. Using the CT setup, data were acquired in five pigs immediately after the injection of 0.05 mmol and 0.07 mmol/kg BW Gd-DTPA. For phantom measurements, mean signal values were determined using a region-of-interest (ROI) analysis and for animal measurements, a ROI was placed in the pulmonary trunk of the cine CT perfusion data sets. The CT phantom measurements were used to calculate the in-vivo maximum CAC corresponding to the HU values obtained in the pulmonary trunk by the cine CT study. Linearity of the AIF of the CT perfusion measurements was verified using the MR phantom measurement results. MR phantom measurements demonstrated linearity for concentrations of 0-4 mM. CT phantom measurements showed linear relation for the entire CAC range. Comparing in-vivo and in-vitro measurements, three of five CA injections at 0.05 mmol/kg and all 0.07 mmol/kg injections exceeded the range of linearity in MRI. The CA dose for quantification of lung perfusion with time-resolved MR studies must be chosen carefully since even with low doses (0.05 mmol/kg) the CAC may exceed the range of linearity in the AIF.     

Pharmacodynamics of streptavidin-coated cyanoacrylate microbubbles designed for molecular ultrasound imaging

OBJECTIVES: To assess the pharmacodynamic behavior of cyanoacrylate, streptavidin-coated microbubbles (MBs) and to investigate their suitability for molecular ultrasound imaging. MATERIALS AND METHODS: Biodistribution of MBs was analyzed in tumor-bearing mice using gamma-counting, immunohistochemistry, flow cytometry, and ultrasound. Further, vascular endothelial growth factor receptor 2-antibody coupled MBs were used to image tumor neovasculature. RESULTS: After 1 minute >90% of MBs were cleared from the blood and pooled in the lungs, liver, and spleen. Subsequently, within 1 hour a decent reincrease of MB-concentration was observed in the blood. The remaining MBs were removed by liver and spleen macrophages. About 30% of the phagocytosed MBs were intact after 48 hours. Shell fragments were found in the kidneys only. No relevant MB-accumulation was observed in tumors. In contrast, vascular endothelial growth factor receptor 2-specific MBs accumulated significantly within the tumor vasculature (P < 0.05). CONCLUSIONS: The pharmacokinetic behavior of streptavidin-coated cyanoacrylate MBs has been studied. In this context, the low amount of MBs in tumors after >5 minutes is beneficial for specific targeting of angiogenesis.     

Molecular Ultrasound Imaging of Early Vascular Response in Prostate Tumors Irradiated with Carbon Ions

Individualized treatments with combination of radiotherapy and targeted drugs require knowledge about the behavior of molecular targets after irradiation. Angiogenic marker expression has been studied after conventional radiotherapy, but little is known about marker response to charged particles. For the very first time, we used molecular ultrasound imaging to intraindividually track changes in angiogenic marker expression after carbon ion irradiation in experimental tumors. Expression of intercellular adhesion molecule-1 (ICAM-1) and of α_vβ₃-integrin in subcutaneous AT-1 prostate cancers in rats treated with carbon ions (16 Gy) was studied using molecular ultrasound and immunohistochemistry. For this purpose, cyanoacrylate microbubbles were synthesized and linked to specific ligands. The accumulation of targeted microbubbles in tumors was quantified before and 36 hours after irradiation. In addition, tumor vascularization was analyzed using volumetric Doppler ultrasound. In tumors, the accumulation of targeted microbubbles was significantly higher than in nonspecific ones and could be inhibited competitively. Before irradiation, no difference in binding of α_vβ₃-integrin-specific or ICAM-1-specific microbubbles was observed in treated and untreated animals. After irradiation, however, treated animals showed a significantly higher binding of α_vβ₃-integrin-specific microbubbles and an enhanced binding of ICAM-1-specific microbubbles than untreated controls. In both groups, a decrease in vascularization occurred during tumor growth, but no significant difference was observed between irradiated and nonirradiated tumors. In conclusion, carbon ion irradiation upregulates ICAM-1 and α_vβ₃-integrin expression in tumor neovasculature. Molecular ultrasound can indicate the regulation of these markers and thus may help to identify the optimal drugs and time points in individualized therapy regimens.