Hg1, novel peptide inhibitor specific for Kv1.3 channels from first scorpion Kunitz-type potassium channel toxin family.

The potassium channel Kv1.3 is an attractive pharmacological target for autoimmune diseases. Specific peptide inhibitors are key prospects for diagnosing and treating these diseases. Here, we identified the first scorpion Kunitz-type potassium channel toxin family with three groups and seven members. In addition to their function as trypsin inhibitors with dissociation constants of 140 nM for recombinant LmKTT-1a, 160 nM for LmKTT-1b, 124 nM for LmKTT-1c, 136 nM for BmKTT-1, 420 nM for BmKTT-2, 760 nM for BmKTT-3, and 107 nM for Hg1, all seven recombinant scorpion Kunitz-type toxins could block the Kv1.3 channel. Electrophysiological experiments showed that six of seven scorpion toxins inhibited ~50-80% of Kv1.3 channel currents at a concentration of 1 µM. The exception was rBmKTT-3, which had weak activity. The IC(50) values of rBmKTT-1, rBmKTT-2, and rHg1 for Kv1.3 channels were ~129.7, 371.3, and 6.2 nM, respectively. Further pharmacological experiments indicated that rHg1 was a highly selective Kv1.3 channel inhibitor with weak affinity for other potassium channels. Different from classical Kunitz-type potassium channel toxins with N-terminal regions as the channel-interacting interfaces, the channel-interacting interface of Hg1 was in the C-terminal region. In conclusion, these findings describe the first scorpion Kunitz-type potassium channel toxin family, of which a novel inhibitor, Hg1, is specific for Kv1.3 channels. Their structural and functional diversity strongly suggest that Kunitz-type toxins are a new source to screen and design potential peptides for diagnosing and treating Kv1.3-mediated autoimmune diseases.

Is Weisskoff model valid for the correction of contrast agent extravasation with combined T1 and T2* effects in dynamic susceptibility contrast MRI?

PURPOSE: The Weisskoff model has been widely applied for correcting the T1 effect of the contrast agent leakage in the measured dynamic susceptibility contrast (DSC)-MRI signals. This study aimed to modify the Weisskoff model for the inclusion of both T1 and T2 effects of the contrast agent extravasation. METHODS: A two-compartment model was proposed and implemented into the original Weisskoff model to describe the combined T1 and T2 effects from the contrast agent leakage in the measured DSC-MRI signals. A computer simulation was performed to evaluate the dependence of T, versus T2 dominance on imaging parameter, field strength, baseline T1, and severity of the leakage. The modified Weisskoff model was employed to correct the relative cerebral blood volume (rCBV) maps in three patients with brain tumors to demonstrate its use. RESULTS: The resultant equation had the same mathematical form as the original model, but with a different expression for the fitting constant K2. This new parameter can be of either a positive or a negative value. Results of the computer simulation showed more probable T2 dominance with longer TE, higher field strength, shorter baseline T1, and greater extraction of the contrast agent. Clinical data were well fitted by the model, with a positive K2 indicating T1 dominance and underestimated rCBV and a negative K2 indicating T2 dominance and overestimated rCBV. The K2 values of normal-appearing brain tissues were distributed in a much smaller range than the K2 values of enhancing tumors. The ratios of corrected over uncorrected normalized CBV (nCBV) for gray matter (GM) were in the range between 1.04 and 1.05, meaning that the nCBV remained rather stable before and after correction. The ratios for the tumors were 0.65, 0.42, and 2.81, either much smaller or greater than the ratios for GM. CONCLUSIONS: This study proposed a modified Weisskoff model that was able to explain both T1 and T2 dominant effects of the contrast agent extravasation in DSC-MRI. Further development is needed to make the K2 parameter a quantitative indicator of the vessel permeability.

Vessel size imaging using dual contrast agent injections.

PURPOSE: To investigate the feasibility of a vessel size imaging (VSI) technique with separate contrast agent injections for evaluation of the vessel caliber in normal tissues and in brain tumors. MATERIALS AND METHODS: Computer simulation was first performed to assess the potential errors in the estimation of vessel caliber that could result from time shifts between the dual contrast agent injections. Eight patients (four female, four male, 37-77 years old) with brain tumors (three high-grade gliomas, two low-grade gliomas, and three meningiomas) were recruited for clinical study. Dynamic susceptibility contrast magnetic resonance imaging (MRI) using gradient echo (GE) and spin echo (SE) echo-planar imaging sequences were performed separately with a 10-minute interval on a 3.0T scanner. Vessel caliber maps were calculated and analyzed in regions of interest at cortical gray matter (GM), thalamus, white matter (WM), and tumors. RESULTS: From the computer simulation, the error of vessel caliber measurement was less than 8% when the difference between the time-to-peak of the GE and the SE studies was 1.5 seconds, and reduced to within 5% when the difference was 1 second. From the patient datasets of a 64 x 64 matrix, the estimated vessel calibers were 37.4 +/- 12.9 microm for cortical gray matter, 20.7 +/- 8.8 microm for thalamus, and 15.0 +/- 5.1 microm for white matter, comparable to results in the literature. Two patients had a VSI with 128 x 128 matrix and showed similar results in vessel calibers of normal tissues. All the tumors had larger mean vessel diameter than normal-appearing tissues. The difference in vascular size between normal tissue and tumor was demonstrated clearly in both the VSIs of regular and high spatial resolution. CONCLUSION: This study suggests that VSI with a dual injection method is a feasible technique for estimating microvascular calibers of normal tissues and brain tumors in clinical scanners.