One-day protocol for imaging of the nigrostriatal dopaminergic pathway in Parkinson's disease by [123I]FPCIT SPECT.
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Parkinson's disease is characterized by degeneration of dopaminergic neurons, resulting in loss of dopamine transporters in the striatum. Recently, the tracer 1231-N-omega-fluoropropyl-2beta-carbomethoxy-3beta-(4-iodoph enyl)nortropane (FPCIT) was developed for imaging dopamine transporters with SPECT. The purpose of this study was to develop an [123I]FPCIT SPECT protocol for routine clinical studies. METHODS: We examined the time course of [123I]FPCIT binding to dopamine transporters in 10 healthy volunteers and 19 patients with Parkinson's disease. RESULTS: We found that the time of peak specific striatal [123I]FPCIT binding was highly varied among subjects, but specific binding peaked in all controls and patients within 3 h postinjection. Between 3 and 6 h, the ratio of specific-to-nonspecific striatal [123I]FPCIT binding was stable in both groups, although, as expected, it was significantly lower in patients. In the patients, [123I]FPCIT binding in the putamen was lower than in the caudate nucleus, and contralateral striatal binding was significantly lower than ipsilateral striatal binding. The subgroup of patients with hemi-Parkinson's disease showed loss of striatal dopamine transporters, even on the ipsilateral side. CONCLUSION: For routine clinical [123I]FPCIT SPECT studies, we recommend imaging at a single time point, between 3 and 6 h postinjection, and using a tissue ratio as the outcome measure. The [123I]FPCIT SPECT technique is sensitive enough to distinguish control subjects from patients with Parkinson's disease, even at an early stage of the disease. (+info)
Assessment of myocardial viability using 123I-labeled iodophenylpentadecanoic acid at sustained low flow or after acute infarction and reperfusion.
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123I-labeled iodophenylpentadecanoic acid (IPPA) is a synthetic fatty acid that may be useful for determination of myocardial viability. We investigated the uptake and clearance kinetics of this tracer in canine models of ischemia and infarction. METHODS: In protocol 1, 185 MBq (5 mCi) 123I-IPPA were injected intravenously in 19 dogs with 50% left anterior descending artery (LAD) flow reduction. In 9 dogs, 201TI was coinjected. In protocol 2, 5 dogs underwent LAD occlusion for 3 h, and 123I-IPPA was injected 60 min after reperfusion. All dogs had flow measured by microspheres, regional systolic thickening by ultrasonic crystals and measurements of postmortem risk area and infarct size. Tracer activities were quantified by gamma well counting and by serial imaging. RESULTS: In protocol 1 dogs with sustained low flow (50% +/- 4%) and absence of systolic thickening (-3.2% +/- 1%), 123I-IPPA defect magnitude (LAD/left circumflex artery [LCX] count ratios) decreased from 0.65 +/- 0.02 to 0.74 +/- 0.02 at 30 min and to 0.84 +/- 0.03 at 2 h (P < 0.01), indicative of rest redistribution. Final transmural 123I-IPPA LAD/LCX activity ratio (0.99 +/- 0.05) was significantly greater than the flow ratio (0.53 +/- 0.04) at injection, confirming complete rest redistribution. The final 123I-IPPA activity ratio was significantly greater than the 201TI ratio over the 2-h period (P < 0.01). In protocol 2 dogs that underwent 3 h of total LAD occlusion and reflow (infarct size = 51% +/- 13% of risk area), viability was overestimated with 123I-IPPA, because uptake averaged 64% of normal in the central necrotic region, where flow averaged < 10% of normal. CONCLUSION: These findings suggest that serial 123I-IPPA imaging may be useful for assessing myocardial viability under conditions of sustained low flow and myocardial asynergy, such as appears to exist in patients with chronic coronary artery disease and depressed left ventricular function. In contrast, 123I-IPPA given early after reperfusion following prolonged coronary occlusion overestimates the degree of viability and therefore may not provide useful information pertaining to the degree of myocardial salvage after reflow in the setting of acute myocardial infarction. (+info)
Favorable effects of glycolate conjugation on the biodistribution of humanized antiTac Fab fragment.
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One of the major limitations of using intact immunoglobulins for targeting tumors is poor penetration into tissues. Although Fab fragments have been used because of their improved kinetics, they have undesirable high renal accumulation. In this study we tested a new approach to block renal accumulation of Fab. METHODS: We conjugated humanized antiTac Fab fragments, which are directed against the interleukin-2 receptor, with glycolate. The biodistribution, pharmacokinetics and catabolism of glycolated Fab (glyco-Fab) were evaluated at two different levels of substitution (heavy and light) compared with nonglycolated Fab in Tac-antigen-positive (ATAC4) and -negative (A431) tumor-bearing nude mice. The mice received coinjections of 125I-labeled glyco-Fab (3 microCi/1 microg) and 131I-labeled nonglycolated Fab (5 microCi/1 microg). In addition, groups of mice receiving these reagents were also coinfused with 50 mg L-lysine. RESULTS: Significantly less glyco-Fab than nonglycolated Fab accumulated in the kidney (21 versus 189 %ID/g; P < 0.001). A higher proportion of glyco-Fab was excreted into the urine in its intact form. The glyco-Fab survived longer in circulation than nonglycolated Fab. The peak tumor accumulation of glyco-Fab was 2.3-fold greater than that of nonglycolated Fab. Furthermore, the ATAC4 tumor-to-normal tissue ratio of glyco-Fab was much higher in all organs than that of nonglycolated Fab. The heavily glyco-Fab accumulated less in the kidney than the lightly glyco-Fab. The coinjected lysine reduced the renal accumulation of both nonglycolated Fab and glyco-Fab. CONCLUSION: Glyco-Fab is a promising agent because of its lower renal accumulation, higher tumor uptake and higher tumor-to-normal tissue ratio. (+info)
No evidence of altered in vivo benzodiazepine receptor binding in schizophrenia.
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Deficits in gamma-amino-butyric acid (GABA) neurotransmitter systems have been implicated in the pathophysiology of schizophrenia for more than two decades. Previous postmortem and in vivo studies of benzodiazepine (BDZ) receptor density have reported alterations in several brain regions of schizophrenic patients. The goal of this study was to better characterize possible alterations of the in vivo regional distribution volume (VT) of BDZ receptors in schizoprenia, using the selective BDZ antagonist [123I]iomazenil and single photon emission computerized tomography (SPECT). Regional BDZ VT was measured under sustained radiotracer equilibrium conditions. The reproducibility and reliability of this measurement was established in four healthy volunteers. No differences in regional BDZ VT were observed between 16 male schizophrenic patients and 16 matched controls. No relationships were observed between BDZ VT and severity of psychotic symptoms in any of the regions examined. In conclusion, this study failed to identify alterations of BDZ receptors density in schizoprenia. If this illness is associated with deficits in GABA transmission, these deficits do not substantially involve BDZ receptor expression or regulation. (+info)
Mechanism of the upregulation of erythropoietin-induced uptake clearance by the spleen.
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Repeated administration of recombinant human erythropoietin (rhEPO) causes upregulation of receptor-mediated tissue uptake by the spleen (Kato, M., H. Kamiyama, A. Okazaki, K. Kumaki, Y. Kato, and Y. Sugiyama. J. Pharmacol. Exp. Ther. 283: 520-527, 1997). To discover whether such upregulation is due to an increase in target cells, the numbers of colony-forming unit erythroids (CFU-E) and burst-forming unit erythroids (BFU-E), the precursor of CFU-E, were measured in the spleen after rhEPO treatment. The uptake clearance of 125I-labeled rhEPO by the spleen was almost proportional to the number of CFU-E, suggesting that the upregulation is due to an increased number of CFU-E. When growth cells were metabolically labeled with [3H]thymidine in vivo, the radioactivity in bone marrow fell significantly after rhEPO treatment, whereas that in the spleen increased significantly. A cell-fractionation study using Percoll revealed that the radioactivity in the BFU-E fraction of splenic cells increased initially after rhEPO treatment, followed by an increase in radioactivity in the CFU-E fraction with a concomitant reduction in radioactivity in the BFU-E fraction. These results demonstrate that EPO stimulates the migration of BFU-E from bone marrow to spleen, followed by its differentiation into CFU-E in the spleen. In conclusion, the upregulation observed in the spleen is due to its stimulatory effect on the migration of BFU-E. (+info)
Enrichment of transiently transfected mesangial cells by cell sorting after cotransfection with GFP.
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Early passage mesangial cells, like many other nonimmortalized cultured cells, can be difficult to transfect. We devised a simple method to improve the efficiency of transient protein expression under the transcriptional control of promoters in conventional plasmid vectors in rat mesangial cells. We used a vector encoding modified green fluorescent protein (GFP) and sterile fluorescence-activated cell sorting (FACS) to select a population consisting of >90% GFP-expressing cells from passaged nonimmortalized cultures transfected at much lower efficiency. Only 10% transfection efficiency was noted with a beta-galactosidase expression vector alone, but cotransfection with GFP followed by FACS and replating of GFP+ cells yielded greater than fivefold enrichment of cells with detectable beta-galactosidase activity. To demonstrate the expression of a properly oriented and processed membrane protein, we cotransfected GFP with a natriuretic peptide clearance receptor (NPR-C) expression vector. Plasmid-dependent cell surface NPR-C density was enhanced by 89% after FACS, though expression remained lower in selected mesangial cells than in the CHO cell line transfected with the same vector. We conclude that cotransfection of rat mesangial cells with GFP, followed by FACS, results in improvement in transient transfection efficiencies to levels that should suffice for many applications. (+info)
125I-Radioimmunoassay of amikacin and comparison with a microbioassay.
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A radioimmunoassay (RIA) has been developed using 125I-amikacin. Amikacin was iodinated by a modified BOLTON and HUNTER method. Dextran-charcoal was used to separate bound from free drug. The standard curve was linear on a logit-log plot in the range of 0.5 ng to 4 ng amikacin per tube. There was no cross-reactivity of amikacin antisera to the amino-glycosides gentamicin, tobramycin, netilmicin, and sisomicin but a 70% cross-reaction was observed with kanamycin, the compound from which amikacin is synthetically derived. Correlation of the RIA with a microbioassay for the determination of serum amikacin levels in 18 patient samples was excellent (r = 0.94). This new RIA technique is more sensitive, rapid, versatile, and less costly than the RIA using 3H-amikacin, and is far more sensitive and faster than microbioassay. (+info)
Topography of the synaptosomal membrane.
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The composition and disposition of the constituent polypeptides of rat cerebral cortical synaptosomal membranes were analyzed on SDS acrylamide gels. Of 20 bands readily detected, 11 account for greater than 93% of the total protein analyzed. These are: (molecu25); 3 (175); 4 (doublet, 137); 5 (doublet, 97); 6 (68); 7 (61); 8 (54); 9 (44); 10 (37); and 11 (33). Bands 5 and 8-10 are the most prominent and account for greater than 60% of the protein mass or 0.67 of its molecular fraction. By lactoperoxidase iodination, the bulk of the proteins in bands 3, 5, 6, and 8 and a portion of band 11 appear to be located on the external (junctional) face of the membrane of intact synaptosomes; proteins in bands 1, 2, 7, 9, and 10 appear to be localized on the internal (synaptoplasmic) face and become labeled only when synaptosomes are lysed. Further confirmation of the topographical distribution is provided by evidence that bands 3-6, 8, and 11 contain glycoproteins susceptible to labeling in intact synaptosomes by oxidation with galactose oxidase or periodate followed by reduction with NaB3H4. Evidence is provided for significant contributions by tubulin- and actin-like molecules to bands 8 and 9, respectively, suggesting that a substantial fraction of the tubulin in the synaptosomal membrane is disposed externally (accessible to iodination) whereas most, if not all, of the actin appears to exhibit the opposite topography. Similar though weaker inferences can also be drawn with regard to the location of tropomyosin and troponin. Preliminary evidence is provided that postsynaptic densities exhibit a protein and iodination profile distinct from that of the synpatosomal membrane. (+info)