Quantitative assessment of right ventricular structural abnormalities by right ventricular polar mapping of single photon emission computed tomogram.

The identification of right ventricular (RV) abnormalities is clinically important in the evaluation of arrhythmogenic substrates in right ventricular-originated ventricular tachycardia (RVT). The purpose of this study was to determine the diagnostic benefit of quantitative analysis in RV single photon emission computed tomography (SPECT) imaging with (99m)Tc-tetrofosmin/sestamibi in patients with RVT. Thirty patients with RVT (15 with idiopathic RVT and 15 with arrhythmogenic right ventricular cardiomyopathy (ARVC)) were compared with 27 control subjects (including 11 with right bundle branch block) with regard to the semiquantitative RV uptake score in each of six segments and the quantitative RV extent score in polar coordinate map displays by SPECT imaging. The RV total score and RV extent score were compared with the RV global function. Perfusion abnormalities were more frequently detected (P = 0.0001) in the ARVC group (59/90, 65.6%) than in the idiopathic RVT group (4/90, 4.4%) or controls (1/162, 0.6%). The RV extent score in the ARVC group (53.0 +/- 24.8) was significantly higher than that in the idiopathic RVT group (8.4 +/- 10.1) or controls (1.2 +/- 4.9). The RV extent score correlated well with the regional RV perfusion score (P < 0.0001) and with the RV ejection fraction (P < 0.0001). Non-invasive RV perfusion mapping using a (99m)Tc-labelled tracer is useful for the quantitative evaluation of RV substrates in patients with ARVC.

Validation of quantitative gated single photon emission computed tomography and an automated scoring system for the assessment of regional left ventricular systolic function.

Despite its ability to quantify regional perfusion and function, there is no established method for quantification of regional perfusion and function by myocardial gated single photon emission computed tomography (SPECT). The aim of this study was to establish a quantitative index for regional perfusion and systolic function assessment using gated SPECT. Myocardial SPECT was performed at rest using (99m)Tc sestamibi with 8-frame gating in 62 consecutive patients. In addition to computation of left ventricular ejection fraction (LVEF), a new computerized method for quantifying, displaying and automatically grading regional data was developed. Regional function was quantified as wall motion, regional EF, and imaged based, count based, and normalized per cent wall thickenings (%WTs). Regional perfusion was assessed as a relative per cent peak count. Data were displayed on a 25-segmented polar map and automatically graded with a 5-point scale, and then summed scores were calculated. These quantitative parameters were compared to data from radionuclide ventriculography (RNV) and contrast left ventriculography. Gated SPECT had high reproducibilities for calculating global and regional ejection fractions and %WT indices (r=0.811-0.984, P<0.0001), but measurement of wall motion was less reproducible (r=0.555, SEE=7.9, P<0.011). LVEF estimated by gated SPECT and summed perfusion scores correlated closely (P<0.0001) with angiographic LVEF. Among the summed function indices that correlated closely with LVEF, normalized %WT had the closest correlations with LVEF estimated by RNV (r=0.657, P<0.0001) and by gated SPECT (r=0.778, P<0.0001). Assessment by visual reviewing of cine-mode playback or by normalized %WT had greater overall sensitivity, specificity, and positive and negative predictive values for detecting impaired regional function among the functional parameters: 71%, 79%, 63% and 84% for cine format analysis, and 78%, 73%, 59% and 87% for normalized %WT, respectively. Thus, besides LVEF, quantitative gated SPECT can provide reproducible and reliable quantitative data on regional perfusion and function. Automated summed scores obtained by gated SPECT can reflect integrated abnormalities of regional perfusion and function of the left ventricle. Both visual analyses by cine-mode display and a functional map of normalized wall thickening have greater diagnostic values for detecting regional function deficit related to coronary artery disease.

A family with liddle's syndrome caused by a mutation in the beta subunit of the epithelial sodium channel.

Liddle's syndrome is a rare form of autosomal-dominant salt-sensitive hypertension. Constitutive activation of the amiloride-sensitive distal renal epithelial sodium channel (ENaC) is essential for salt-sensitive hypertension. Recently, several DNA analysis studies have indicated that there is a mutation of C-terminus of either the beta or y subunit. We sequenced the C-termini of the beta and -gamma subunits of the ENaC in a Japanese family with hypertension and hypopotassemia without excess minerarocorticoids, clinically diagnosed as Liddle's syndrome. The mutation of the ENaC of this family was beta R564X. Since such case seem to be rare in the literature, detailed data are shown in this report.

Cardiac imaging in a patient with anomalous origin of the left coronary artery from the pulmonary artery--a case report.

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is an uncommon congenital heart disease and has a high mortality rate in infancy. However, myocardial ischemia does not develop until adolescence or adulthood in about 10% of patients. Moreover, the diagnosis of ALCAPA is often difficult in cases without heart murmur or cardiac symptoms. The authors report the case of a 31-year-old man with ALCAPA. He was admitted to the hospital for evaluation of mild shortness of breath at exercise, but he had no typical chest symptoms due to myocardial ischemia or heart failure until age 31 and he had no heart murmur. Moreover, electrocardiogram did not show an old myocardial infarction or myocardial ischemia. Therefore, the authors did not suspect ALCAPA until they performed transthoracic echocardiography and exercise-stress single photon emission computed tomography (SPECT) with Tc-99m-tetrofosmin. The final diagnosis was established from the results of coronary arteriography. In the present case, a transthoracic echocardiogram showed abnormal coronary circulation, and exercise-stress SPECT revealed reversible myocardial ischemia. Transthoracic echocardiography and myocardial SPECT imaging could be a useful noninvasive tools for diagnosing the ALCAPA.

Impaired myocardial accumulation of 15-(p-iodophenyl)-9-(R,S)-methylpentadecanoic acid in a patient with hypertrophic cardiomyopathy and exercise-induced ischemia due to vasospasm.

We encountered a patient with hypertrophic cardiomyopathy complicated with exercise-induced myocardial ischemia. Exercise-stress 99mTc-tetrofosmin imaging demonstrated reversible ischemia in the lateral wall, whereas resting fatty acid imaging with a new beta-methyl branched fatty acid analogue, I-123-15-(p-iodophenyl)-9-(R,S)-methylpentadecanoic acid (123I-9-MPA), showed impaired uptake and accelerated washout kinetics in the inferoapical and posteroseptal walls but not in the ischemia-related region. These findings suggest that the metabolic derangement is closely related to cardiomyopathy per se rather than exercise-induced myocardial ischemia in this patient with hypertrophic cardiomyopathy and a spastic coronary lesion so that myocardial perfusion and 123I-9-MPA imagings may contribute to clarifying the etiological background of impaired myocardial fatty acid metabolism.

[A case of labial adhesions with urinary retention].

A 69-year-old woman was admitted to our hospital complaining of urinary retention. On examination, the labia was found to be fused extensively, with a pinhole opening at the midline. The adhesions were separated under spinal anesthesia. One year after operation, there remains no recurrence. Etiological factor in labial adhesions are thought to include a combination of local inflammation and estrogen deficiency. In our case, the patient had hip joint disease. Hip joint disease is shown to influence the formation of adhesions by interfering with perineal hygiene and decreasing sexual activity.

[Relationship of fetal hind limb and fetal liver on amino acid metabolism in pregnant rats].

A tracer technique for injections into rat fetal abdomen in utero was employed to obtain a better knowledge of amino acid metabolism in fetal hind limb and fetal liver. Calculation of tissue fluid distribution via 3H-inulin space made possible an estimation of the fetal hind limb and fetal liver intracellular amino acid concentration based upon the fetal plasma and fetal tissue amino acid concentration. A significant concentration gradient between fetal plasma and fetal hind limb was found for glutamate (37.3) but not for alanine (9.7) or leucine (3.0). The radioactivity of fetal hind limb and fetal liver after the injection of 1 microCi of radioactive 14C-glutamate 14C-alanine or 14C-leucine into fetal abdomen was measured. In fetal hind limb, significant radioactivity was recognized after 14C-alanine administration, but not after 14C-glutamate administration as against the significant concentration gradient. However, in fetal liver, significant radioactivity was recognized after 14C-glutamate administration, but not after 14C-alanine administration. We concluded that in fetus, glutamate was released from the hind limbs and a large amount taken up by fetal liver.

The study of placental L-ascorbate (vitamin C) transport mechanism (using microvillous membrane vesicles).

To investigate the placental L-ascorbate (Vitamin C) transport mechanism, the uptake of L-ascorbate into microvillous membrane vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-ascorbate into microvillous membrane vesicles was osmotically sensitive. This finding indicated that the uptake of L-ascorbate into microvillous membrane vesicles represented transport into the vesicles. The uptake of L-ascorbate into microvillous membrane vesicles was not dependent on sodium electrochemical gradient. The initial rate of uptake was not changed when the intravesicular space was rendered electrically more negative by membrane diffusion potential induced by the use of highly permeant anions. The initial rate of L-ascorbate transport exhibited saturation kinetics with respect to the L-ascorbate concentration; an apparent Km of 1.33 mM and Vmax of 47p mol/mg protein/20 sec was calculated. The uptake of L-ascorbate into microvillous membrane vesicles was competitively inhibited by D-isoascorbate. These results indicated that transport of L-ascorbate across the placental microvillous membrane vesicles was carrier mediated and was passive transport.

[Human placental glutathione transport mechanism].

The placental transport mechanism of glutathione (GSH) was investigated using microvillous membrane vesicles prepared from human term placenta. Using (3H-glycine)-labeled-GSH, it was clarified that GSH in the extravesicular compartment of placental microvillous membranes was rapidly degraded by gamma-GTP (gamma-glutamyltranspeptidase) and resulting amino acid, and 3H-labeled-glycine was actively transported via a sodium cotransport system. AT-125 treated microvillous membrane vesicles almost entirely lost its gamma-GTP activity, and showed intact GSH transport. Using AT-125 treated microvillous membrane vesicles, it was revealed that GSH was transported across the microvillous membrane as an anion via a membrane potential-dependent mechanism. These results indicated that gamma-GTP which existed in microvillous membrane played a role in GSH metabolism and that intracellular GSH was translocated out of the syncythiotrophoblast cell into the maternal blood space via a specific carrier in microvillous membrane because the GSH concentration was higher in intracellular than extracellular and extracellular membrane potential was positively charged.

[Nonenzymatic glucosylation of human placental trophoblast basement membrane collagen (relation to diabetic placenta pathology)].

Nonenzymatic glucosylation is a reaction in which glucose binds nonenzymatically to hemoglobin, serum protein and glomerular basement membrane collagen etc. It has been thought that nonenzymatic glucosylation results in functional and chemical changes in those substances (hemoglobin etc) and contributes to the pathological changes in diabetes mellitus. This time we investigated whether nonenzymatic glucosylation occurred in human placental trophoblast basement (TrBM) collagen or not. The ability of glucose to interact with TrBM collagen (nonenzymatic glucosylation of TrBM collagen) was examined by incubating TrBM collagen with 3H-D-glucose in vitro. As a result it was shown that nonenzymatic glucosylation occurred in TrBM collagen and nonenzymatic glucosylation of TrBM collagen depended on the glucose concentration, reaction time and reaction temperature. These results indicate that possibly hyperglycemia, via nonenzymatic glucosylation modifies the function and chemistry of TrBM collagen and is related to the placental pathological changes in diabetic pregnancy.

[The transport mechanism of antibiotics using microvillous membrane vesicles (placental transport of fosfomycin)].

Using the rapid filtration technique, the uptake of fosfomycin into microvillous membrane vesicles isolated from human term placental trophoblast was investigated. The microvillous membrane vesicles exhibited the uptake of fosfomycin into an osmotically reactive intravesicular space and it was indicated that the uptake of fosfomycin by microvillous membrane vesicles represented transport into membrane vesicles. The uptake of fosfomycin by microvillous membrane vesicles was not dependent on the Na+ electrochemical gradient or membrane potential. The initial uptake of fosfomycin by microvillous membrane vesicles did not exhibit saturation kinetics with respect to fosfomycin concentration, and increased linearly as the fosfomycin concentration increased. These results indicated that fosfomycin was transported across the microvillous membrane by simple diffusion. L-alanine, L-valine, L-lysine, inorganic phosphate or D-glucose did not inhibit the uptake of fosfomycin into microvillous membrane vesicles. On the other hand, fosfomycin did not inhibit the uptake of L-alanine, L-valine, L-lysine inorganic phosphate or D-glucose into microvillous membrane vesicles. These results revealed that fosfomycin did not affect the placental transport activity of other nutrients.

[Study on the mechanism of placental transport of L-lysine (using human placental microvillous (brush border) membrane vesicles)].

The uptake of L-lysine in human placental microvilli vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-lysine into the vesicles was osmotically sensitive. This finding indicates that the uptake of L-lysine by placental microvilli vesicles represents transport into the membrane vesicles. The uptake of L-lysine into microvilli vesicles is sodium independent. The initial rate of uptake is markedly increased when the intravesicular space is rendered electrically more negative by membrane diffusion potential induced by the use of highly permeant anions or by K+ diffusion potentials via valinomycin. These results indicate that the sodium independent uptake of L-lysine into the microvilli membrane vesicles is dependent on the electrical potential difference of the membranes. A kinetic analysis of the uptake demonstrated that two transport systems for vesicular entry of L-lysine existed with a Km1 of 0.13 mM, Vmax1 of 590 pmol/mg protein/20 sec, Km2 of 0.91 mM, Vmax2 of 2010 pmol/mg protein/20 sec. The uptake of L-lysine in microvilli vesicles was inhibited by dibasic amino acid (L-arginine, L-ornithine, L-glutamine), but not by other classes of amino acid. These results indicate the existence of a dibasic amino acid specific transport system in placental microvilli membrane.

[Studies on L-glutamate transport mechanism in human placental trophoblast microvilli membrane vesicles].

The uptake of L-glutamate in brush border (microvilli) vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-glutamate into the vesicles occurred osmotically, and preincubation with L-glutamate increased the uptake of amino acid. These findings indicate that the uptake of L-glutamate by placental trophoblast brush border membranes represents the transport into membrane vesicles. A Na+ electrochemical gradient (extravesicular greater than intravesicular) stimulated the initial rate of L-glutamate uptake about three times. The initial rate of transport exhibited saturation kinetics with respect to the L-glutamate concentration; an apparent Km of 0.15 mM and V max of 70 pmol/mg protein in 20 seconds were calculated. The uptake of L-glutamate into the vesicles was competitively inhibited by L-glutamate and L-cysteate (acidic amino acid). These results indicate that a Na-dependent acidic amino acid specific transport system exists in the placental trophoblast microvilli membrane. These results indicate that the transport of L-glutamate across the placental microvilli membrane is sodium-dependent and carrier mediated.

[Placental transport of taurine in brush border microvilli].

Placental transport of taurine was studied in isolated brush border microvillous plasma membrane vesicles by a rapid filtration technique. Brush border microvillous plasma membrane vesicles were prepared from syncytio trophoblast of human term placenta by a method of differential centrifugation and calcium precipitation. The specific activities of alkaline phosphatase, 5' nucleotidase and gamma-GTP in the membrane preparation were enriched to 13-14 times, 12-13 times, and 5-6 times respectively, as high as those in the homogenate. The membrane vesicles exhibit uptake of 3H-labeled taurine into an osmotically reactive intravesicular space. Taurine uptake by vesicles was stimulated specifically by an inward sodium gradient, and replacement of NaCl in the transport medium by KCl, LiCl, and choline chloride had no effect on the transport activity of the vesicles. Taurine transport is inhibited competitively by the presence of beta alanine and GABA. The initial rate of transport followed saturation kinetics with respect to the taurine concentration: An apparent Km of 0.22mM and Vmax of 67 pmol/mg protein were calculated in 20 seconds. These results indicate that transport of taurine across the placental brush border membrane is sodium dependent and carrier mediated.

[Amino acid transfer at the fetal surface of the trophoblast and the characterization of its carrier in the rat placenta in vivo].

A new technique for injecting tracer into the rat fetus in utero, intraperitoneally, was employed to characterize the amino acid exchange at the fetal surface of the trophoblast and to study the transplacental amino acid transfer. Calculation of tissue fluid distribution via 3H-inulin space made it possible to estimate the placental intracellular amino acid concentration on the basis of the cord and maternal plasma concentration. The fetal plasma free amino acid was taken up at a considerable rate by the fetal surface of the placenta in umbilical circulation. The amino acid from fetal plasma was utilized for the protein synthesis and deamination in the placenta. It was speculated that the amino acid transfer at the fetal surface of the placenta was carrier-mediated facilitated diffusion, not simple diffusion, on the basis of the data showing the stereospecificity for the isomers. From these results it was considered that the fetal, not maternal, plasma free amino acid concentration greatly influenced the fetal amino acid net uptake.

Effect of cyclic AMP on placental active transport of amino acid in rats.

In pursuit of the probable relationship of cAMP to the placental active transport of amino acids as suggested by our previous human data, experiments were done in this study with dibutyryl cAMP and theophylline loaded in pregnant rats to examine the changes in level of 14C-lysine in the maternal liver, placenta and fetus. The results are: (1) Loading of dibutyryl cAMP had no effect on the uptake of 14C-lysine in the maternal liver, placenta or fetus, except that the correction of the values with its concentration in the maternal blood revealed only a slightly significant elevation in the value in each of them 30 min after the loading. (2) Loading of theophylline significantly increased the uptake of 14C-lysine in all these organs. (3) The content of cAMP in the placenta significantly increased with the loading of theophylline. These results have led the authors to believe that endogenous cAMP is significantly linked with the placental active transport of amino acids, though there is no apparent contribution to it by exogenous cAMP.