Microwave-stimulated fixation for preembedding immunoelectron microscopy.
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Fixatives↗
Biomedical subjects
H Ohtani
Publications and source records attributed to H Ohtani.
[Comparison of 99mTc-hexakis 2-methoxy-2-isobutyl isonitrile and 201Tl SPECT imaging for evaluation of coronary artery disease].
To evaluate clinical value of 99mTc-hexakis 2-methoxy-2-isobutyl isonitrile (MIBI) imaging for assessing coronary artery disease (CAD), 99mTc-MIBI SPECT imaging at post-exercise and at rest was compared with 201Tl SPECT imaging at post-exercise and 3 hours redistribution in 27 patients suspected with CAD. The sensitivities for detecting CAD patients were 94% (17/18) by both studies. The specificities were 71% (5/7) by 99mTc-MIBI and 57% (4/7) by 201Tl (p = NS). The sensitivities for detecting stenosed coronary arteries (greater than or equal to 75% stenosis) were also similar between 99mTc-MIBI (78%) and 201Tl (74%) (p = NS). The similar specificity values were obtained by 99mTc-MIBI (84%) and by 201Tl (82%) (p = NS). The patterns of abnormality (normal, ischemia and scar) were similar between 99mTc-MIBI and 201Tl images in 22 of the 25 cases (88%) and 117 of the 125 segments (94%). However, these patterns were occasionally different particularly in patients who received PTCA or CABG. Thus, 99mTc-MIBI SPECT imaging seems to be as accurate as 201Tl SPECT imaging for the detecting and evaluating CAD.
Adult↗
[Comparison of interobserver variance between 99mTc-MIBI and 201Tl SPECT].
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Adult↗
Prediction of reversible ischemia after coronary artery bypass grafting by positron emission tomography.
Metabolic imaging using positron emission tomography (PET) facilitates the identification of ischemic but viable myocardium. In this study, the predictive value of PET for identifying improvement in regional function after coronary artery bypass grafting (CABG) was assessed. PET perfusion and metabolic imagings using N-13 ammonia and F-18 deoxyglucose (FDG) were performed before and 5-7 weeks after CABG in 25 patients with coronary artery disease. Each of the 5 myocardial segments of the left ventricle was categorized as normal, ischemic and infarcted based on the findings of perfusion and PET metabolic images. Among 58 hypoperfused segments, abnormal perfusion in 17 of 25 ischemic segments was correctly predicted to be reversible (68% prediction accuracy), and that in 25 of 33 infarcted segments were correctly predicted to be irreversible (76% prediction accuracy) (p < 0.001). Similarly, among 53 asynergy segments assessed by radionuclide ventriculography, abnormal wall motion in 21 of 27 asynergy segments was correctly predicted to be reversible (78% prediction accuracy), and that in 21 of 26 PET viable segments was correctly predicted to be irreversible (81% prediction accuracy) (p < 0.001). Thus, preoperative metabolic imaging using PET appears to be useful for predicting responses to CABG.
Adult↗
[Minimal redistribution of thallium-201 representing reversible ischemia after coronary bypass surgery: value of quantitative analysis of exercise thallium-201 SPECT].
We previously reported the value of minimal redistribution (MR) of thallium-201 in analyzing quantitative polar map analysis (QNT) which correlated well with metabolic activity on FDG-PET. To determine whether ischemic areas that have redistribution are truly reversible, we performed stress and 3-hr delayed thallium-201 SPECT imaging and radionuclide ventriculography (RNV) in 41 patients with coronary artery disease before and after coronary bypass surgery (CABG). Redistribution (RD) was categorized into 4 grades: complete RD (CR), incomplete RD (IR), MR and persistent defect (PD). MR was defined as the segment that showed > or = 2SD improvement in more than 1/3 of the area on QNT, but RD was not evident by visual analysis. 1. QNT identified MR in 30 of 56 segments (54%) where PD is noted by visual inspection. 2. The MR segments showed severer wall motion abnormality (wall motion score: 1.83 +/- 1.12) than did the IR or CR segments (0.99 +/- 1.04) (p < 0.01), but the wall motion was well preserved, compared to the PD segments (2.54 +/- 0.90) (p < 0.01). 3. The grade of RD was compared with improvement in regional perfusion and wall motion on RNV after CABG. Improvement in perfusion was observed in 62 of 77 IR or CR segments (81%) and in 17 of 30 MR segments (57%), but in only 3 of 26 PD segments (12%) (p < 0.01). 4. Similarly, improvement in wall motion was observed in 45 of 57 IR or CR segments (79%) and in 22 of 27 MR segments (81%), but in only 5 of 25 PD segments (20%) (p < 0.01). Thus, the MR segments should be considered reversible after CABG. We conclude that QNT of RD should be performed to detect RD which is slight, yet suggestive of viability.
Aged↗
[A bull's eye subtraction polar map for assessing thallium redistribution (1). Comparison with quantitative redistribution].
A new bull's eye subtraction polar (BS) map has been developed for quantitative assessment of redistribution on stress-delayed thallium-201 SPECT studies. BS map was created after subtraction of a normalized stress bull's eye polar map from a normalized delayed polar map in 10 normal subjects and 33 patients with coronary artery disease to compare BS map with qualitative redistribution scores. The BS map showed less than 15% in all of the 50 segments in the normal subjects. Of 27 segments exhibiting persistent defect without redistribution, the BS map showed less than 10% in 20 segments (74%) and 10-15% in 6 segments (22%). On the other hand, of 34 segments exhibiting redistribution, the BS map showed greater than or equal to 15% in 25 segments (74%) and 10-15% in the remaining 9 segments (26%). Thus, significant redistribution was evident in the areas showing greater than or equal to 10-15% on the BS map. We conclude that the BS map is valuable for quantitative assessment of redistribution on stress-delayed thallium-201 SPECT imaging.
Adult↗
Metabolic activity in the areas of new fill-in after thallium-201 reinjection: comparison with positron emission tomography using fluorine-18-deoxyglucose.
Reinjection of thallium-201 after recording the 3-hr delayed scan often demonstrates improvement in areas of persistent abnormalities. To determine the metabolic activity of these areas, the changes seen on stress/redistribution/reinjection thallium SPECT were compared with PET using fluorine-18-fluorodeoxyglucose (FDG) in 18 patients with coronary artery disease. Of 48 segments showing no redistribution on the delayed scan, the reinjection scan identified new fill-in in 20 segments (42%), all of which demonstrated FDG uptake. In contrast, only 7 of the 28 segments (25%) showing no fill-in after reinjection were PET viable (p less than 0.01). Eleven patients had coronary bypass graft surgery after the radionuclide study. The majority of the segments showing redistribution (87%) and new fill-in after reinjection (65%) improved in wall motion, whereas only eight segments (25%) without new fill-in improved after surgery. Of those without new fill-in, two segments showing PET ischemia improved in wall motion, whereas the remaining six segments showing PET scar did not improve after surgery. Thus, the segments showing new fill-in after reinjection are PET viable myocardium. However, reinjection thallium imaging still underestimates the extent of tissue viability compared to PET imaging.
Coronary Circulation↗
Regional wall thickening of left ventricle evaluated by gated positron emission tomography in relation to myocardial perfusion and glucose metabolism.
Regional wall thickening was assessed by electrocardiographically gated positron emission tomography (ECG-gated PET) in 26 patients with coronary artery disease. The standardized percent count increase from end-diastole to end-systole (S-percent Cl) was calculated as an index of wall thickening. The S-percent Cl was 77.8% +/- 28.9% in the segments with normal perfusion at rest, 51.9% +/- 29.5% in those with mild hypoperfusion, and 32.8% +/- 30.9% in those with severe hypoperfusion (p less than 0.001, each). Among the segments with resting hypoperfusion, the S-percent Cl was 38.9% +/- 31.5% in those without stress-induced ischemia and 48.7% +/- 30.9% in those with ischemia (p less than 0.05). Furthermore, among resting severe hypoperfusion, the S-percent Cl was 23.0% +/- 23.9% in the segments without fluorine-18-fluorodeoxyglucose (FDG) uptake and 37.8 +/- 32.9% in those with FDG uptake (p less than 0.05). These results suggest that stress-induced ischemia and FDG accumulation correlated with wall thickening. Thus, quantitative analysis of regional wall thickening seems to be useful for combined analysis of regional function, perfusion and metabolism in coronary patients.
Adult↗
Value of thallium-201 reinjection after delayed SPECT imaging for predicting reversible ischemia after coronary artery bypass grafting.
The reinjection of a small dose (40 MBq) of thallium-201 after stress and delayed imaging often shows new redistribution in the regions with persistent defect. To assess whether these segments may represent reversible ischemia, reinjection thallium-201 single-photon emission computed tomography (SPECT) was performed after stress and 3-hour delayed imaging in 24 patients before coronary artery bypass grafting (CABG). The left ventricular myocardium was divided into 5 myocardial segments and regional wall motion was scored on a scale from 0 (normal) to 4 (dyskinesia). Thallium-201 findings were compared with improvement in regional perfusion and wall motion 1 to 2 months after CABG. The reinjection imaging identified new redistribution in 15 of 32 persistent defects (47%) on the 3-hour delayed images. In the study of stress and delayed SPECT imaging, the improvement in perfusion was observed in 34 of 43 segments (79%) exhibiting redistribution and 15 of 32 (47%) segments without redistribution (p less than 0.01). The reinjection SPECT identified new redistribution in 12 of the 15 improved segments that were not detected on the delayed images. Similarly, the improvement in wall motion was observed in 23 of 31 segments (74%) exhibiting redistribution and 14 of 30 segments (47%) without redistribution on the delayed images (p less than 0.05). The reinjection identified new redistribution in 10 of the 14 improved segments that were undetected on the delayed images. The predictive values for improvement in perfusion and wall motion by the reinjection imaging were significantly higher (92 and 89%) than those by the delayed imaging (69 and 62%, respectively, p less than 0.05 each).(ABSTRACT TRUNCATED AT 250 WORDS)
Aged↗
A case of biopsy-proven myocarditis progressing to autopsy-proven dilated cardiomyopathy.
A 22-year-old man presented with congestive heart failure following flulike symptoms. The diagnosis of acute myocarditis was confirmed by endomyocardial biopsy, which revealed mild infiltration of inflammatory cells. A favorable response to beta-adrenergic receptor blockade was seen, and the patient was discharged without symptoms. Five months later, however, congestive heart failure recurred, and intracardiac thrombi were demonstrated. The patient died after two months. Postmortem examination revealed left ventricular dilatation with slight interstitial fibrosis; the diagnosis was dilated cardiomyopathy. Thus, progression of biopsy-proven myocarditis to dilated cardiomyopathy 10 months after the onset of disease was documented.
Adrenergic beta-Antagonists↗
Differing microvasculature in the two major types of gastric carcinoma: a conventional, ultrastructural and ultrastructural immunolocalization study of von Willebrand factor.
The microvasculature of the stroma of human gastric carcinoma was studied by immuno-electron microscopy for factor VIII/von Willebrand factor (vWF) and conventional electron microscopy. In differentiated type (intestinal) gastric carcinoma (9 cases), capillaries were distributed more densely around carcinoma cell nests. vWF was localized in endothelial cells and neighbouring stroma. Ultrastructurally, capillary endothelial cells showed considerable hypertrophic changes with well-developed rough endoplasmic reticulum (rER). vWF was localized in well-developed rER, granules, Weibel-Palade bodies (WPB), in the vascular lumen as clusters, and diffusely deposited in the subendothelium. This indicates that endothelial cells in this group are transformed into a state of active protein production. In undifferentiated type (diffuse) gastric carcinoma (12 cases), capillaries were uniformly distributed and endothelial hypertrophic changes were less remarkable. vWF was localized in WPB, scanty rER and subendothelial matrix. Solid capillary buds were observed in both types; they were composed of a solid strand of endothelial cells without a visible lumen. Our results reveal that the microvasculature in tumour stroma differs significantly according to its histological type.
Capillaries↗
Immunoelectron microscopic localization of human type III collagen in human gastrointestinal carcinomas.
Ultrastructural immunolocalization of human type III collagen/procollagen was investigated in 28 cases of human gastrointestinal carcinoma by the pre-embedding method using a monoclonal antibody. In addition to immunoreactivity on collagen fibers, amorphous or finely fibrillar immunoreactive substances were observed in the extracellular matrix adjacent to the plasma membrane and in the cisternae of rough endoplasmic reticulum (rER). These substances were considered to represent type III procollagen and/or collagen before fiber formation. Cytoplasmic recesses containing immunoreactive fibers were also observed. These findings related to collagen production were detected in stromal fibroblasts in 16 of 28 cases and in carcinoma cells in six of 28 cases. The positivity rate of these findings was lower in diffuse-type gastric carcinoma. The present study thus clarified further details of collagen production and secretion, and revealed that carcinoma cells may also be involved in the production of stromal collagen in some cases as well as stromal fibroblasts.
Antibodies, Monoclonal↗
Immunoelectron microscopic localization of aromatase in human placenta and ovary using microwave fixation.
In this study we investigated the immunohistochemical localization of a unique aromatase, a single protein of 51,000 daltons, in the human placenta and ovary at light and electron microscopic levels. Microwave fixation was adopted for the immunoelectron microscopic study because it is an excellent method for preserving antigenicity and subcellular structures in frozen sections. Tissue samples from four immature human placentas, four full-term human placentas, and two human ovaries fixed in 10% formalin were examined by light microscopy. In addition, tissues from three full-term human placentas and one immature human placenta fixed in 4% paraformaldehyde were examined by electron microscopy. By light microscopy, immunoreactivity for this aromatase was located in the syncytiotrophoblast and a part of the cytotrophoblast of the placenta and in the lutein and granulosa cells of the ovary. Immunoelectron microscopy revealed that the aromatase antigen was localized on the surface of the microvilli, the lateral plasma membrane, and in the endoplasmic reticulum (ER) in the syncytiotrophoblast of the placenta. The positive immunoreactivity in the syncytiotrophoblast ER is consistent with previous results using antibodies for other types of aromatase, whereas the reactivity on the microvilli has not been previously described. The present report describes the fine localization of this unique aromatase in placental and ovarian tissues; its localization on the plasma membranes requires further physiological investigation.
Adult↗
Kinetic studies on drug disposition in rabbits. II. Dose dependent pharmacokinetics of sulfamethizole.
In order to evaluate dose-dependent sulfamethizole (SMZ) kinetics, 100, 300 and 1000 mg of SMZ was constantly infused over 5 min in rabbits and thereafter plasma and urine samples were collected at convenient intervals. When a dose of 100 mg was given, the time course of total plasma concentration followed a biexponential characteristic. For higher doses, plasma decay curves revealed a convex descending feature after the distribution phase. The respective unbound fraction in plasma (fp) at the total plasma concentrations of 200 and 100 micrograms/ml were 0.41 and 0.19. The corresponding total body clearances were 2.6 and 2.2 l/h, indicating that the drug elimination did not contribute to the convex-descending plasma curves. A physiologically based pharmacokinetic model was adapted to various tissue levels of SMZ. No saturable tissue binding was observed and the apparent volume of distribution of SMZ at steady state with a rabbit of 3.3 kg, Vss (1), calculated by tissue volumes and partition coefficients of tissue to unbound plasma was expressed as follows: Vss = 0.14 + 1.86fp. From this relationship, it was shown that the apparent volume of distribution of SMZ was significantly affected by the unbound fraction in plasma and the dose-dependent kinetics after intravenous administration was due to the decrease of the apparent volume of distribution with time. The tissue distribution study contributed significantly to the understanding of the dose-dependent drug kinetics.
Animals↗
Kinetic studies on drug disposition in rabbits. III. Effect of tolbutamide on renal excretion of sulfamethizole.
The effect of tolbutamide (TB) on the urinary excretion of sulfamethizole (SMZ) under constant infusion of SMZ at 100 mg/h was studied. Intravenous administration of TB (50 mg/kg) caused a decrease in the urinary excretion rate of SMZ but an increase in the unbound concentration of SMZ in plasma. The total concentration of SMZ in plasma decreased rapidly after TB injection and then increased gradually to a level higher than the control. The slope of the terminal phase of the unbound concentration of TB in plasma in the presence of SMZ was significantly smaller than that of TB alone. The analysis using the perfusion limited model showed that the elimination kinetics of SMZ in the presence of TB could be described by the mutual displacement of plasma protein binding of both drugs and the competitive inhibition of the tubular secretion of SMZ by the unbound concentration of TB in renal vein. Further the inhibitor constant was in good agreement with that for the in vitro uptake by renal cortex slices.
Animals↗
Kinetic studies on drug disposition in rabbits. I. Renal excretion of iodopyracet and sulfamethizole.
In order to quantify the renal handling of iodopyracet (IOD) and sulfamethizole (SMZ), single-drug clearance studies in rabbits were performed under quasi-steady state conditions with stepwise increasing the infusion rate of IOD or SMZ. Although concentration dependence of plasma protein binding was observed for both drugs, the urinary excretion rate of IOD was proportional to its total plasma concentration at low total plasma concentrations of 0.05-0.8 mM. On the other hand, the relationship between urinary excretion rate and total plasma concentration of SMZ was a concave-ascending curve at low plasma concentrations and the renal clearance of SMZ was sensitive to changes in plasma protein binding. However, renal clearances referenced to unbound plasma concentration at total plasma concentrations of 0.05 mM for IOD and SMZ were 9.5 and 38 l/h, respectively. Those values were much greater than the effective plasma flow in rabbits. These facts indicated that the intrinsic clearances at the sites of tubular secretion were high and that the rates of secretion were fully or partially limited by the renal plasma flow. Furthermore it was suggested that unbound drug was liberated from plasma protein at the sites of tubular secretion. The data obtained at high plasma concentrations indicated that the tubular secretion of IOD had capacity limited characteristics and that the urinary excretion of SMZ involved tubular reabsorption as well as saturable tubular secretion. From the data obtained, a perfusion-limited pharmacokinetic model was constructed characterizing the excretory processes, namely, glomerular filtration, passive tubular reabsorption, saturable tubular secretion and reequilibrium between bound and unbound drugs in plasma. For both drugs, the estimates for bulk flow rate were reasonable values of effective renal plasma flow and the dissociation constants for tubular secretion agreed well with those for in vitro renal cortex accumulation, suggesting that the kinetic model based on physiological concepts was useful for the understanding of the drug elimination processes.
Animals↗
The effect of estrogen and progesterone on the volume-pressure curve of the uterine ampulla in late pregnant rats.
Volume-pressure curves of the uterine ampullae, taken on day 17 of pregnancy from intact pregnant and ovariectomized, vehicle- or estradiol and/or progesterone-treated pregnant rats, during infusion of Ca-free Tyrode's solution containing EGTA at a constant rate were determined, and the effects of the steroids on the Vz value, which was obtained by extrapolating the linear part of the curve to zero pressure, and compliance were evaluated. Both the parameters examined were decreased by ovariectomy. Treatments with estradiol or progesterone alone did not but the steroids combined restored Vz to the level of intact pregnant rats, and partially restored compliance. The significance of the development of Vz by estrogen and progesterone for the maintenance of pregnancy is discussed.
Animals↗
Ultrastructural characterization of enterochromaffin-type cells in human gastric carcinoma.
Ultrastructural characterization has been performed on eosinophilic granular cells occurring in gastric carcinomas. Of these, the present paper dealt with enterochromaffin (EC)-type cells. EC-type cells were identified by the presence of eosinophilic granules, immunoreactivity for serotonin (5 HT) and argentaffinity. Conventional electron microscopy revealed that they bore membrane bound, electron-dense granules of varying size (100-800 nm in shorter diameter), which were larger than those of normal EC cells. Their shapes are round, rod-shaped and concave. The granules are immunolabeled for 5HT by both pre- and post-embedding methods. The preembedding method also disclosed diffuse cytosolic immunoreactivity. Both differentiated-type and undifferentiated-type gastric carcinomas showed essentially the same results. General trends of occurrence of Paneth-like and EC-type cells in gastric carcinomas were also discussed. The present study confirmed that EC-type cells in gastric carcinoma retained basic characteristics of EC cells despite of morphological deviation from the normal cells. It is speculated that they are derived from neoplastic precursor cells with a special differentiation.
Adenocarcinoma↗