Technetium-99m-labeled monoclonal antibodies for immunoscintigraphy. Simplified preparation and evaluation

Ascorbic acid incubated with monoclonal antibodies (22 degrees C, 60 min, pH 6.5) at a molar ratio of 3500:1, reduced 2.7 +/- 0.2% of the available disulfides to sulfhydryl groups that strongly bind 99mTc, and provided greater than 95% labeling efficiency for several IgM, IgG and F(ab')2 antibodies. The colloid formation was consistently less than 3% and the stability of the tracer when challenged with DTPA and cysteine was excellent. The immunospecificity of labeled antibodies as determined by immobilized specific antigen assay was 84 +/- 1% for IgM and 82.6 +/- 1.1% for IgG antibodies. For in vivo evaluation in mice bearing experimental abscesses and tumors, corresponding 125I-labeled antibodies served as controls. The liver uptake was similar (P = 0.76 and P = 0.12) for 99mTc or 125I labeled antinuclear antibody TNT-1 in mice bearing abscesses as well as for 99mTc-TNT-1-F(ab')2 and 125I-TNT-1-F(ab')2 in mice bearing tumors. Higher but statistically insignificant (P = 0.08, 0.18, and 0.73) urinary excretion was noted for 99mTc-antibodies. For corresponding 99mTc- and 125I-labeled antibodies, the abscess to muscle ratios (3.3 +/- 0.5 vs. 3.4 +/- 0.8) and tumor to muscle ratios (10.04 +/- 4.4 vs. 10.54 +/- 3.0) were similar. The high 99mTc-TNT-1-F(ab')2 uptake permitted excellent scintigraphic visualization of tumors whereas the nonspecific 99mTc-HSA did not (tumor/muscle ratio: 2.4 +/- 0.3). This method is simple, reliable, and adaptable to an instant labeling technique.     

Muscle capillary permeability for [14C]inulin and [51Cr]EDTA in human forearm

Capillary permeability of [14C]inulin and [51Cr]EDTA was examined in human forearm in five healthy, subjects by indicator diffusion technique. Injections of, initially [125I]albumin and [14C]inulin, and after 30 min resting, of [125I]albumin and [51Cr]EDTA, were given in a brachial artery. During light exercise of the forearm, blood was sampled in 2-s periods from a deep cubital vein primarily draining muscles. The plasma flow rate, calculated as the dose of [125I]albumin in the injectate divided by the area under the curve for the venous concentration of 125I, was, on average, 8.5 ml min-1 100 g-1 forearm. Assuming [125I]albumin is a partially permeable tracer, a correction for extraction of albumin was performed. This gave extraction fractions of 0.107 +/- 0.015 (mean +/- SEM) for [14C]inulin and 0.377 +/- 0.033 for [51Cr]EDTA, respectively. The capillary permeability surface area product per 100 g tissue (CDC) was for [14C]inulin 0.90 +/- 0.19, and for [51Cr]EDTA 3.31 +/- 0.38 ml min-1 100 g-1 forearm. The average of the ratios of the CDC values of [51Cr]EDTA to those of [14C]inulin, 4.0 +/- 0.5, is significantly higher than the corresponding ratio between the measured free diffusion coefficients in water at 37 degrees C, 3.07 +/- 0.002 (N = 36 and 17, respectively). This indicates that there is some degree of restriction for [14C]inulin (MW 5200) relative to [51Cr]EDTA (MW 340.2) and it points to an 'equivalent pore radius estimate' of about 160 A in human muscle capillaries.     

Generalized five-dimensional dynamic and spectral factor analysis

We have generalized the spectral factor analysis and the factor analysis of dynamic sequences (FADS) in SPECT imaging to a five-dimensional general factor analysis model (5D-GFA), where the five dimensions are the three spatial dimensions, photon energy, and time. The generalized model yields a significant advantage in terms of the ratio of the number of equations to that of unknowns in the factor analysis problem in dynamic SPECT studies. We solved the 5D model using a least-squares approach. In addition to the traditional non-negativity constraints, we constrained the solution using a priori knowledge of both time and energy, assuming that primary factors (spectra) are Gaussian-shaped with full-width at half-maximum equal to gamma camera energy resolution. 5D-GFA was validated in a simultaneous pre-/post-synaptic dual isotope dynamic phantom study where 99mTc and 123I activities were used to model early Parkinson disease studies. 5D-GFA was also applied to simultaneous perfusion/dopamine transporter (DAT) dynamic SPECT in rhesus monkeys. In the striatal phantom, 5D-GFA yielded significantly more accurate and precise estimates of both primary 99mTc (bias=6.4 % +/- 4.3 %) and 1231 (-1.7% +/- 6.9%) time activity curves (TAC) compared to conventional FADS (biases = 15.5% +/- 10.6% in 99mTc and 8.3% +/- 12.7% in 123I, p < 0.05). Our technique was also validated in two primate dynamic dual isotope perfusion/DAT transporter studies. Biases of 99mTc-HMPAO and 123I-DAT activity estimates with respect to estimates obtained in the presence of only one radionuclide (sequential imaging) were significantly lower with 5D-GFA (9.4% +/- 4.3% for 99mTc-HMPAO and 8.7% +/-4.1% for 123I-DAT) compared to biases greater than 15% for volumes of interest (VOI) over the reconstructed volumes (p < 0.05). 5D-GFA is a novel and promising approach in dynamic SPECT imaging that can also be used in other modalities. It allows accurate and precise dynamic analysis while compensating for Compton scatter and cross-talk.     

Thermal modulation of the toxicokinetics of benzo[a]pyrene in isolated hepatocytes of sablefish (Anoplopoma fimbria), black rockfish (Sebastes melanops), and chub mackerel (Scomber japonicus)

Hepatocytes from sablefish (Anoplopoma fimbria), black rockfish (Sebastes melanops) and chub mackerel (Scomber japonicus) were isolated from 11 degrees C acclimated animals. The uptake, metabolism, and excretion of benzo[a]pyrene (B[a]P) in hepatocytes was measured at 6, 11 and 19 degrees C. Chub mackerel hepatocyte uptake rates were significantly lower (0.012 +/- 0.003 microg/s per g cell) at 11 degrees C than black rockfish (0.028 +/- 0.009 microg/s per g cell) or sablefish (0.032 +/- 0.012 microg/s per g cell) hepatocytes at all temperatures. Hepatocytes metabolized B[a]P to phase I (1-8%) and phase II (92-99%) metabolites. Accumulation of phase II metabolites was lower in chub mackerel hepatocytes (0.016 +/- 0.004 microg/h per g cell), than black rockfish (0.052 +/- 0.012 microg/h per g cell), or sablefish hepatocytes (0.060 +/- 0.015 microg/h per g cell). Phase II metabolite accumulation increased greatest with temperature in chub mackerel hepatocytes (Q10 = 1.94 +/- 0.30), followed by sablefish (Q10 = 1.65 +/- 0.30), and rockfish (Q10 = 1.38 +/- 0.30). Sablefish hepatocytes had higher excretion rates of phase II metabolites (0.010 +/- 0.0023 microg/h per g cell), than mackerel (0.0046 +/- 0.0009 microg/h per g cell) or rockfish hepatocytes (0.0029 +/- 0.0008 microg/h per g cell). Phase II metabolite excretion rates increased with temperature only in sablefish hepatocytes (Q1O = 1.67 +/- 0.76). These differences in toxicokinetics may indicate distinct consequences for various species exposed to xenobiotics.     

Striatal Dopaminergic Functioning in Patients with Sporadic and Hereditary Spastic Paraplegias with Parkinsonism

Sporadic spastic paraplegia (SSP) and hereditary spastic paraplegia (HSP) belong to a clinical and genetically heterogeneous group of disorders characterized by progressive spasticity and weakness in the lower extremities. The symptoms are associated with pyramidal tract dysfunction and degeneration of the corticospinal tracts. Parkinsonism is uncommon in SSP/HSP patients. However, both disorders are associated with damage to the nigrostriatal dopaminergic system. In the present study, the clinical features of patients with SSP/HSP were investigated, and nigrostriatal dopaminergic binding potential was assessed using dopamine transporter (DAT) single-photon emission computer tomography (SPECT). Nine patients with spastic paraplegia participated in the present study. The subjects underwent DAT SPECT using the agent [2-[[2-[[[3-(4-chlorophenyl)-8-methyl-8-azabicyclo[3,2,1]oct-2-yl]methyl](2-mercaptoethyl)amino]ethyl]amino]ethanethiolato (3-)-N2,N20,S2,S20]oxo-[IR-(exo-exo)])-[⁹⁹mTc]technetium ([⁹⁹mTc]TRODAT-1). The [⁹⁹mTc]TRODAT-1 SPECT images of five patients appeared normal, whereas the images of four patients revealed reduced striatal ligand uptake. Among the four patients with reduced uptake, two had parkinsonism, and one exhibited periodic limb movements and restless leg syndrome. Our DAT SPECT imaging study shows that reduced DAT density may be observed in patients with parkinsonism. The results of the present study offer an explanation for the spectrum of spastic paraplegia symptoms and the progression of the disorder.     

Estimating gluconeogenesis by NMR isotopomer distribution analysis of [13C]bicarbonate and [1-13C]lactate

The gluconeogenic contribution to glucose production in livers isolated from rats fasted for 24 h was determined by 13C-NMR isotopomer distribution analysis of secreted glucose enriched from 99% [13C]bicarbonate (n = 4) and 99% [1-13C]lactate (n = 4). Experiments with 3% 2H2O were also performed, allowing the gluconeogenic contribution to be measured by the relative 2H enrichments at positions 5 and 2 of glucose. From 13C-NMR analyses, the contribution of gluconeogenesis to glucose output was estimated to be 93 +/- 3% for [13C]bicarbonate perfusion and 91 +/- 3% for [1-13C]lactate perfusion, in good agreement with the 2H-NMR analysis of the gluconeogenic contribution to glucose production (100 +/- 1% and 99 +/- 1%, respectively) and consistent with the expected negligible contribution from glycogenolysis. These results indicate that 13C-NMR analysis of glucose 13C-isotopomer distribution from either [13C]bicarbonate or [1-13C]lactate precursor provides realistic estimates of the gluconeogenic contribution to hepatic glucose output.     

A dynamic phantom for radionuclide renography

The aim of the study was to develop and test a dynamic phantom simulating radionuclide renography. The phantom consisted of five partly lead covered plastic containers simulating kidneys, heart, bladder and background (soft tissues, liver and spleen). Dynamics were performed with multiple movable steel plates between containers and a gamma camera. Control of the plates is performed manually with a stopwatch following exact time schedules. The containers were filled with activities (99mTc) which produce count rates close to clinical situations. Count rates produced by the phantom were compared with ten clinical renography cases: five 99mTc MAG3 and five 99mTc DTPA examinations. Two phantom simulations were repeated three times with separate fillings, acquisitions and analyses. Precision errors as a coefficient of variation (CV) of repeated measurements were calculated and theoretical values were compared with the corresponding measured ones. A multicentre comparison was made between 19 nuclear medicine laboratories and three clinical cases were simulated with the phantom. Correlations between count rates produced by the phantom and clinical studies were r = 0.964 for 99mTc MAG3 (p < 0.001) and 0.961 for 99mTc DTPA (p < 0.001). The precision error was 4.5 +/- 3.2% and the percentage difference between theoretical and measured values for Tmax was 4.0 +/- 1.6%. Images and curves of the scanned phantom were close to a real patient in all 19 laboratories but calculated parameters varied: the difference between theoretical and measured values for Tmax was 6.8 +/- 6.2%. The difference between laboratories is most probably due to variations in acquisition protocols and analysis programs: 19 laboratories with 18 different protocols and 8 different programs. The dynamics were found to be repeatable and suitable for calibration purposes for radionuclide renography programs and protocols as well as for multicentre comparisons.     

Comparison of correction techniques for simultaneous 201Tl/99mTc myocardial perfusion SPECT imaging: a dog study

We compared two correction methods for simultaneous 201Tl/99mTc dual-isotope single-photon emission computed tomography (SPECT). Both approaches use the information from the third energy window placed between the photopeak windows of the 201Tl and 99mTc. The first approach, described by Moore et al, corrects only for the contribution of the 99mTc to the 201Tl primary 70 keV window. We developed the three-window transformation dual isotope correction method, which is a simultaneous cross-talk correction. The two correction methods were compared in a simultaneous 201Tl/99mTc sestamibi cardiac dog study. Three separate acquisitions were performed in this dog study: two single-isotope and one dual-isotope acquisition. The 201Tl single-isotope images were used as references. The total number of counts, and the contrast between the left ventricular cavity (LVC) and the myocardium, were used in 70 keV short axis slices as parameters for evaluating the results of the dual-isotope correction methods. Three consecutive short-axis slices were used to calculate averaged contrast and the averaged total number of counts. The total number of the counts was 667000+/-500 and 414500+/-400 counts for the dual isotope (201Tl+/-99mTc) and single-isotope (201Tl-only) 70 keV images, respectively. The corrected dual-isotope images had 514700+/-700 and 368000+/-600 counts for Moore's correction and our approach, respectively. Moore's method improved contrast in the dual isotope 70 keV image to 0.14+/-0.03 from 0.11+/-0.02, which was the value in the 70 keV non-corrected dual-isotope image. Our method improved the same contrast to 0.22+/-0.03. The contrast in the 201Tl single-isotope 70 keV image was 0.28+/-0.02. Both methods improved the 70 keV dual-isotope images. However, our approach provided slightly better images than Moore's correction when compared with 201Tl-only 70 keV images.     

Fiberglass limb phantoms: fabrication and use for quantitative scintigraphy

Quantitative radionuclide scintigraphy often requires empirical calibration factors derived from phantoms which simulate the radioactivity distribution, tissue geometry and tissue composition of the region of interest. This paper describes a method in which casts made with fiberglass tape of the region of interest. This paper describes a method in which casts made with fiberglass tape are used to form realistic, water-fillable phantoms of the limbs. Phantoms were constructed for the hind legs of the dog and rabbit, species frequently used in developing new radioscintigraphic techniques. Leg bones removed from euthanized animals were mounted anatomically within the casts. The dimensions of the phantom cavities were determined by x-ray computed tomography. A procedure was developed for orienting the phantoms to match the hind leg geometry of a given experimental setup. Use of the phantoms for image activity calibration is illustrated for a geometric-mean counting technique used to determine 99mTc activity densities in soft-tissue regions of the dog thigh. Generalization of the calibration technique to planar and tomographic imaging is straightforward. In situ measurements of 99mTc activity density obtained by external counting were compared with in vitro radioassays of excised tissue. For 22 tissue samples obtained from four dogs, the in situ and in vitro data were linearly correlated (r = 0.98, p much less than or equal to 0.001) over a 50-fold range of activity density. The mean and standard deviation of the observed percent discrepancies [% discrepancy = 100 (in situ - in vitro)/in vitro] were (7.8 +/- 2.9) and (13.7 +/- 2.1), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Permeability of the sheep placenta to unmetabolized polar non-electrolytes.

1. The rate of appearance in uterine venous blood of radioactively labelled, polar, non-electrolytes has been measured following their injection into the foetal circulation in the chronically catheterized sheep near term. Uterine blood flow was measured by an antipyrine technique. 2. Estimates of a placental permeability constant not corrected for area were 61-2 +/- 5-5 ml. min-1 (mean +/- S.E. of mean) for [14C] urea, 1-85 +/- 0-16 for [14C]erythritol and 0-21 +/- 0-03 for [3H]mannitol. Results are also presented for [14C]ethylene glycol, [14C]L-glucose, [14C]mannitol, [51Cr] EDTA and [3H] and [14C]sucrose. 3. In four sheep, permeability measurements for several solutes were made and the results were analysed in terms of restricted diffusion via cylindrical, water-filled pores. Calculation of pore radius was made by a minimum variance method and values ranging from 0-43 to 0-45 nm were obtained. 4. Stability and absence of protein binding of probe molecules was investigated by gel permeation on Sephadex columns.     

Blood flow, histomorphology and elemental composition of the canine femur after physical training or immobilization

Effects of long-term physical training (treadmill running 4 km/day for 15 weeks) and immobilization (casting of the limb for 11 weeks) on the blood flow, bone marrow uptake of radio-colloid, histomorphology as well as elemental composition of the proximal canine femur were investigated. In the runners both bone marrow and mean bone perfusions measured with a 133Xe washout method were increased by 53.5% (P less than 0.05) and 54.1% (P less than 0.05), respectively, as compared with the age-matched controls. Bone marrow uptake of [99mTc]tincolloid and volumetric fractions of trabecular bone and bone marrow were not significantly altered after training or immobilization. The amount of zinc measured by atomic absorption spectrophotometry was, however, significantly higher (P less than 0.01) after immobilization as compared with the controls. Consequently, long-term physical training is suggested to stimulate bone metabolism by enhancing its blood perfusion even in the resting state, whereas the increase of zinc content was considered to be linked to the obvious bone resorption caused by immobilization.     

Pulmonary clearance of inhaled [99Tcm]DTPA: effects of ventilation pattern

While a rise in lung volume is known to increase the pulmonary clearance of technetium-99m-labelled dietylene triamine pentaacetate ([99Tcm]DTPA), little interest has been focused on the effects of changes in ventilation frequency, tidal volume and airway pressure. We studied adult, anaesthetized and intubated rabbits during three ventilation patterns (VP) using pressure controlled ventilation (ServoVentilator 900C). VP was either deep slow (f = 20 min-1, tidal volume (VT) = 30 +/- 4 ml kg-1 and positive end-expiratory pressure (PEEP) = 0.2 kPa [VP 20/0.2, n = 8]) or rapid shallow (f = 80 min-1, VT = 11 +/- 2 ml kg-1 and PEEP = 0.2 or 0.4 kPa [VP 80/0.2, n = 6 and VP 80/0.4, n = 6]). The mean airway pressure was similar at VP 20/0.2 and VP 80/0.4. During administration of [99Tcm]DTPA aerosol all animals were ventilated under the same conditions (f = 40 min-1 and PEEP = 0.2 kPa). The pulmonary clearance rate expressed as the half-life time (T1/2) of [99Tcm]DTPA was at VP 80/0.2 = 113 +/- 31 min, at VP 80/0.4 = 70 +/- 24 min (P less than 0.01 compared to VP 80/0.2) and at VP 20/0.2 = 36 +/- 18 min (P less than 0.001 compared to VP 80/0.2 and P less than 0.01 compared to VP 80/0.4). We conclude that the pulmonary clearance of [99Tcm]DTPA increases (1) during rapid shallow ventilation when PEEP is increased from 0.2 to 0.4 kPa; (2) during deep slow ventilation relative to rapid shallow ventilation even when the mean airway pressure is similar.     

重组人骨形成蛋白-2-珊瑚人工骨复合物在拔牙窝牙槽骨修复中的作用

目的:探讨重组人骨形成蛋白-2与珊瑚人工骨复合物(复合骨 )在拔牙窝修复中的作用。方法:拔除 12只成年狗两侧上颌第二及第三切牙,并去除牙槽窝之间的牙槽间隔,一侧随即植入复合骨,对侧植入珊瑚人工骨(珊瑚骨 )作为对照。并于植入后 4、8、12周取材,采用组织学观察、图像分析和 [⁹⁹Tc^m]-MDP核素骨显像等方法比较两种植入材料在牙槽窝中的骨修复能力。结果:复合骨植入牙槽骨后,材料被逐渐降解吸收,新骨不断生成,12周后,植入材料完全被成熟的骨组织取代;图像分析结果显示不同时间复合骨组新骨形成的比值显著高于珊瑚骨组(P <0.05 );4和 8周复合骨组核素浓聚程度高于珊瑚骨组,12周两组核素浓聚程度差异不明显。结论:复合骨在牙槽骨缺损中的骨修复能力和修复效果明显优于珊瑚骨.     

甲状腺切除术后应用甲状腺平面显像评估残余甲状腺:与治疗剂量131I扫描的对比

目的:我国常用^99mTc甲状腺扫描评估甲状腺癌术后残余甲状腺大小,本文拟对甲状腺扫描与治疗剂量的¹³¹I扫描进行比较分析。方法:148例（男51,女97）2年内就诊甲状腺癌术后病人入组,年龄（47.0±13.3）岁,2名核医学科医师独立、盲法阅片,采用5分法计分。统计学分析采用秩相关研究,相关系数采用t检验进一步比较。结果:69/148（46.6%）两种显像结果得分一致,79（53.4%）¹³¹I扫描得分高于甲扫。甲扫及¹³¹I扫描评分结果与甲状腺激素水平均相关（P<0.001）,且相关系数间甲扫与T₃、T₄相关程度更高（t₁=-1.933,P=0.028;t₂=-2.788,P=0.003）。结论:^99mTc甲状腺平面显像可作为术后残余甲状腺的评估方法,但¹³¹I扫描显示范围更大,两者与甲状腺激素水平均相关,但甲扫与其相关性可能更高。     

Determination of postglomerular permselectivity to neutral dextrans in the dog

The single-pass multiple-indicator-dilution (MID) technique was used to analyze postglomerular capillary permeability. Anesthetized mongrel dogs (n = 13) during mannitol diuresis received a pulse injection of 125I-albumin (plasma reference), [14C]inulin (glomerular reference), creatinine (interstitial reference), and a homogeneous [3H]dextran molecular weight marker 6,000-12,000 dalton in the left renal artery. Simultaneous left renal venous outflow and right and left urine were rapidly sampled. Left urine recoveries of creatinine, [14C]inulin, and [3H]dextran were identical, indicating no glomerular solute flux limitation. Progressive precession of the [14C]inulin and [3H]dextran renal vein curves relative to creatinine indicated increasing postglomerular limitation to solute flux proportional to molecular size. The postglomerular solute extraction (EPG) (renal vein upslope indicator/125I-albumin) varied inversely with postglomerular renal plasma flow (F), indicating diffusion limitation. Ouabain infusion into the left renal artery significantly reduced Na reabsorption but did not alter the EPG of [14C]inulin or [3H]dextran. Permeability-surface (PS) area products calculated from EPG and F ranged from 4.86 +/- 0.89 to 0.97 +/- 0.25 (SD) cm X s-1 X 100 g-1 for indicators 5,000-12,000 dalton. [14C]Inulin PS products remained constant for F greater than or equal to 2.50 ml X s-1 X 100 g kidney-1. PS[3H]dextran/PS[14C]In (n = 20, F greater than or equal to 2.5 ml X s-1) was used to calculate an effective postglomerular capillary pore radius, r = 55.5 +/- 7.6 (SD) A.     

Effect of advanced age on p-aminohippurate-induced inhibition of renal tubular secretion in male Fischer 344 rats

The effect of aging on glomerular filtration, effective renal plasma flow and on the responsiveness of the renal tubular anion secretory system to inhibition by 4-aminobenzoylglycine (p-aminohippurate, PAH) was examined in young (5-month) and old (22-month) Fischer 344 male rats. Plasma clearance, protein binding and renal extraction of [131I]o-iodohippurate, [125I]iothalamate and HPLC-purified [99mTc]mercaptoacetyltriglycine (MAG3), were used as in vivo probes of renal function. The effect of advanced age, without concomitant PAH, on the disposition of these markers was initially determined in ketamine anesthetized, temperature-maintained male rats, ages 5, 14 and 22 months by means of constant infusion clearance studies. Aging per se decreased (P less than 0.05) the kidney-weight normalized or body weight-normalized GFR and effective renal plasma flow rates. GFR values averaged 1.67, 1.43 and 1.32 ml/min per g kidney for the 5-, 14- and 22-month-old rats, respectively. Kidney- or body weight-normalized clearances of MAG3 and o-iodohippurate showed similar (25-27%) decreases, whereas the absolute values (ml/min) for GFR, o-iodohippurate and MAG3 clearance rates were not altered by aging. The effective filtration fraction, extraction ratio and plasma protein binding were also unchanged by advanced age. Overall, the age-related decreases in renal function were minimal in Fischer-344 rats, compared to other species. Differences in data normalization, species and gender account, in part, for discrepancies observed when comparing results in different studies on the effects of advanced age on renal function. Subsequently, we examined the effect of aging on the renal responsiveness to inhibition of tubular anion secretion using constant rate PAH infusion studies, adjusted for age-related changes in renal function. Aging did not alter PAH-induced inhibition of iodohippurate secretion. Inhibition of MAG3 elimination was more pronounced in the old rats compared to the young controls.     

Evaluation of an in vitro endothelialized vascular graft under pulsatile shear stress with a novel radiolabeling procedure

OBJECTIVE: To improve the hemocompatibility of vascular grafts, endothelial cell (EC) seeding of biomaterials prior to implantation is critical. The current in vitro study was designed to investigate such a feasibility on a collagen-coated heparin-bonded graft and to evaluate cell detachment upon pulsatile shear stress. MATERIALS AND METHODS: Endothelial cells (EA-hy-926) were seeded onto grafts. The endothelialization of the grafts was evaluated by the [3H]-thymidine incorporation, scanning electron microscopy (SEM) and histological examinations. After in situ EC radiolabeling with a novel 99mTc technique, the prostheses were exposed to pulsatile shear stress (0.27 N/m2), mimicking the shear rate occurring in a superficial femoral artery, for 3 h in a flow circuit and EC loss quantified by gamma camera detection. RESULTS: Complete EC coverage was achieved after 5 days. Three hours of artificial perfusion resulted in a low EC loss (12.9+/-0.8%, n = 7). SEM shows EC withstanding shear stress in valleys of prosthesis circumvolutions. CONCLUSIONS: These satisfactory results could be explained by the high affinity of EC for heparinized surfaces in addition to cell surface receptors involved in adhesion to collagen.     

Investigation of relationship between idiopathic hypercalciuria and urinary enzyme activities

In patients with uncomplicated idiopathic hypercalciuria renal function is normal except for increased renal calcium excretion. In this study, the level of fractional urinary enzyme excretion was assessed in relation to calciuria. Fourteen patients with a mean age of 5.8 +/- 0.8 years who had daily urinary calcium excretion more than 4 mg/kg and with otherwise normal renal function tests were included in the study. None of the patients manifested either renal calculus or nephrocalcinosis. Fourteen normal children with a mean age of 5.4 +/- 0.74 were included in the control group. The level of the urinary N-acetyl beta-D glucosaminidase (NAG) to creatinine ratio, fractional aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) excretion were not significantly different compared to the control group (p > 0.05). The patients were subdivided according to the type of hypercalciuria. The levels of NAG/creatinine ratio, fractional ALT, AST, ALP, LDH excretion were not significantly different in the absorptive type of calciuria group compared to the control group (p > 0.05). In conclusion, hypercalciuria during childhood which is 6.46 +/- 1.83 mg/kg/day is not related to the levels of NAG/creatinine ratio, fractional ALT, AST, ALP and LDH excretion in urine.     

Blockade of long-term potentiation by beta-amyloid peptides in the CA1 region of the rat hippocampus in vivo

The effect of intracerebroventricular (icv) injections of beta-amyloid peptide fragments Abeta[15-25], Abeta[25-35], and Abeta[35-25] were examined on synaptic transmission and long-term potentiation (LTP) in the hippocampal CA1 region in vivo. Rats were anesthetized using urethan, and changes in synaptic efficacy were determined from the slope of the excitatory postsynaptic potential (EPSP). Baseline synaptic responses were monitored for 30 min prior to icv injection of Abeta peptides or vehicle. High-frequency stimulation (HFS) to induce LTP was applied to the Schaffer-collateral pathway 5 min or 1 h following the icv injection. HFS comprised 3 episodes of 10 stimuli at 200 Hz, 10 times, applied at 30-s intervals. Normal LTP measured 30 min following HFS, was produced following icv injection of vehicle (191 +/- 17%, mean +/- SE, n = 6) or Abeta[15-25; 100 nmol] (177 +/- 6%, n = 6) 1 h prior to HFS. LTP was, however, markedly reduced by Abeta[25-35; 10 nmol] (129 +/- 9%, n = 6, P < 0.001) and blocked by Abeta[25-35; 100 nmol] (99 +/- 6%, n = 6, P < 0.001). Injection of the reverse peptide, Abeta[35-25], also impaired LTP at concentrations of 10 nmol (136 +/- 3%, n = 6, P < 0.01) and 100 nmol (144 +/- 7, n = 8, P < 0.05). Using a different protocol, HFS was delivered 5 min following Abeta injections, and LTP was measured 1 h post HFS. Stable LTP was produced in the control group (188 +/- 15%, n = 7) and blocked by Abeta[25-35, 100 nmol] (108 +/- 15%, n = 6, P < 0.001). A lower dose of Abeta[25-35; 10 nmol] did not significantly impair LTP (176 +/- 30%, n = 4). The Abeta-peptides tested were also shown to have no significant effect on paired pulse facilitation (interstimulus interval of 50 ms), suggesting that neither presynaptic transmitter release or activity of interneurons in vivo are affected. The effects of Abeta on LTP are therefore likely to be mediated via a postsynaptic mechanism. This in vivo model of LTP is extremely sensitive to Abeta-peptides that can impair LTP in a time- ([25-35]) and concentration-dependent manner ([25-35] and [35-25]). These effects of Abeta-peptides may then contribute to the cognitive deficits associated with Alzheimer's disease.