Minimal deviation endometrioid adenocarcinoma of the endometrium and its MRI findings.

Minimal deviation endometrioid adenocarcinoma (MDA-E) of the endometrium is a rare pathological entity, and its radiological features are rarely documented. A 73-year-old Japanese woman was referred to the authors when an endometrial biopsy revealed moderately differentiated endometrioid adenocarcinoma. Preoperative radiological examinations, including ultrasound, computed tomography (CT), and magnetic resonance imaging (MRI) showed no evidence of cancer nests. In the hysterectomy specimen, mildly atypical glands were scattered throughout the entire depth of the myometrium, without stromal desmoplastic reaction, and a tiny focus of typical, ruptured, endometrioid adenocarcinoma glands was found in the atrophic endometrium. MRI had not been able to identify this unusual, scattered, myometrial invasion. It should be kept in mind that in cases showing Stage IA endometrial carcinoma without endometrial thickening on MRI, this rare form of invasion may be present.

RF generation using a compact bench gyromagnetic line.

The search for new technologies aiming to reach radiofrequency (RF) generation in different manners for diverse ends is a constant demand for several applications. The goal is to develop cost-effective and simpler systems compared to those that already exist. Our motivation is to reach an alternative way of generating RF in pulsed transmission systems employing a gyromagnetic nonlinear transmission line (GNLTL). The GNLTL consists of a ferrite-loaded-coaxial transmission line and can produce a large frequency spectrum with RF conversion efficiency above 10% from about 200 MHz up to the frequency of 2-4 GHz (S-band) for potential space-based applications. In a GNLTL, the signal amplitude is related to its propagation velocity since the peak voltage travels faster than its portion of lower amplitudes since the ferrite permeability decreases with the current amplitude. As the pulse crest travels faster than its valley, a time reduction happens in the output rise time, called pulse sharpening. Besides, the magnetic moments of ferrite dipoles initially aligned with the axial magnetic bias are displaced from their original position by the azimuthal field generated around the inner conductor by the current pulse, resulting in a damped precession movement. This movement happens along the line length as the current pulse propagates, inducing high-frequency oscillations. In short, the paper's goal is to present the experimental results using a 60-cm gyromagnetic line to provide RF in the GHz range using a solenoid for magnetic bias on a testing bench. Finally, the paper discusses the influence of the azimuthal and the axial magnetic fields on the output signal with the ferrite rings operating in a saturation state during the current pulse propagation.

Epidural puncture can be confirmed by the Queckenstedt-test procedure in patients with cervical spinal canal stenosis.

BACKGROUND: The loss-of-resistance test is the most popular method for identifying the epidural space, but it cannot confirm epidural puncture. Therefore, we developed a new method to confirm epidural puncture by assessing indirect changes in epidural pressure using the Queckenstedt-test procedure, which increases subarachnoid pressure by compressing the internal jugular veins. Because this new method depends on the dynamics of cerebrospinal fluid, blockade of cerebrospinal fluid flow, as with severe spinal stenosis, is predicted to reduce changes in epidural pressure. Thus, in this study, we examined the effect of spinal stenosis on the Queckenstedt-test procedure. METHODS: Epidural puncture using the loss-of-resistance test was utilized to insert an electrode in patients undergoing cervical spine surgery. Epidural pressure was monitored during bilateral compression of the internal jugular veins to confirm epidural puncture. The insertion of the electrode into the epidural space was confirmed by observation of muscle twitch evoked by electric stimulation. RESULTS: In 60 patients, epidural puncture was performed with the loss-of-resistance test; a second trial was required in 13 patients. Increased epidural pressure was observed in 57/73 trials. When increased epidural pressure was observed, epidural puncture was always successful. The sensitivity and specificity of this method was 92.0% and 100%, respectively. The positive and negative predictive values were 100% and 66.7%, respectively. CONCLUSION: An increase in epidural pressure during bilateral compression of the internal jugular veins could offer a reliable method for confirming epidural puncture in combination with the loss-of-resistance test, even if patients have potential spinal canal narrowing.

Bleeding time prolonged by daily low-dose aspirin is shortened by one medium dose aspirin.

BACKGROUND: In planning surgery, a low-dose aspirin regimen for prevention of thrombotic events is often discontinued in order to avoid the risk of excessive bleeding during surgery. However, this procedure increases the risk from adverse thrombotic events. We propose a different method, which may normalize the prolonged bleeding time caused by low-dose aspirin. We verified the effectiveness of this method in healthy volunteers. METHODS: Volunteers with bleeding time prolonged by taking 81 mg of aspirin a day for a period of 1 week were randomly divided into two groups. The test group of 18 volunteers received a dose of 660 mg of aspirin, while the control group of 16 received placebo. Bleeding time and maximum platelet activity were then evaluated. RESULTS: Before 660 mg of aspirin or placebo, bleeding time was prolonged: in the aspirin group from 3.1 +/- 0.7 to 6.1 +/- 1.4 min (n=18), and in the placebo group from 2.9 +/- 0.9 to 6.1 +/- 1.5 min (n=16). This prolongation was significant in both groups at the P<0.01 level. In the test group, bleeding time was shortened to 4.5 +/- 1.3 min (P<0.01), which is in the normal range, while it remained prolonged in the control group (6.0 +/- 1.2 min). Platelet activity, on the other hand, was suppressed in both groups. CONCLUSION: We conclude that 660 mg of aspirin effectively shortens the bleeding time prolonged by daily low-dose (81 mg) aspirin.

Reduction of radiation-induced apoptosis by specific expression of Bcl-2 in normal cells.

Radiation-induced apoptosis is thought to underlie the toxicity of radiation to normal tissues as well as to cancer cells. We hypothesized that specific ectopic overexpression of the antiapoptotic molecule Bcl-2 in normal cells would inhibit radiation-induced apoptosis and thereby reduce radiation-induced toxicity in normal cells. To express Bcl-2 specifically in normal cells (which have wild-type (wt) p53) but not in cancer cells (which often have mutated p53), we constructed a Bcl-2 expression plasmid (PG13-Bcl-2) with a minimal promoter regulated by multiple wt p53 DNA-binding sites and found that the presence of wt p53 protein strongly upregulated Bcl-2 expression through this plasmid. Transfection of NIH 3T3 fibroblasts, which express wt p53, with PG13-Bcl-2 increased cell survival and reduced apoptosis; however, transfection of MDA-MB-231 breast cancer cells, which have mutated p53, did not affect survival and apoptosis of those cells. These results indicate that irradiation of normal cells rapidly upregulates the expression of wt p53, which binds to the p53 binding sequence of the PG13-Bcl-2 plasmid and increases the transcriptional activity of Bcl-2. Ectopic expression of Bcl-2 reduced radiation-induced apoptosis only in normal cells (not in cancer cells). Bcl-2 expression was detected in the lung from mice injected via a tail vein with LPD-PG13-Bcl-2 or LPD-CMV-Bcl-2, but did not in the lung from mice treated with DOTAP or LPD-PG13-mock. This novel approach to inhibiting radiation-induced apoptosis in normal cells may allow such cells to be protected from radiation-induced toxicity. Further preclinical in vivo studies are needed.

Association between arterial stiffness and platelet activation.

Increased arterial stiffness is strongly associated with atherosclerosis, while platelet activation is an important trigger of thrombotic events in patients with atherosclerosis. However, little is known about the effect of arterial stiffness on platelet activation. We therefore investigated the association between arterial stiffness and platelet activation in 38 normal volunteers (20 men and 18 women) aged 23-77 years (mean = 49 +/- 15 years). Arterial stiffness was assessed by measuring brachial-ankle pulse wave velocity (ba-PWV) and heart-brachial PWV (hb-PWV). Flow cytometric analyses were performed to evaluate platelet activation by measuring surface expression of P-selectin and platelet-neutrophil complexes (PNC) before and after activation by ADP. We also calculated the difference between basal and stimulated states of P-selectin and PNC to assess platelet activation reserve. PWVs were significantly correlated with age and BP (r = 0.60-0.81). For platelet activation and activation reserve, correlations with age were less strong but remained significant (r = 0.36-0.61), with the exception of P-selectin (not significant, NS), and correlations with SBP were similar (r = 0.35-0.53). A significant correlation was found between PWVs and platelet activation (r = 0.43-0.74). Multiple regression analysis demonstrated significant correlations between platelet activation and reserve and PWVs (coefficient = 2.17-6.59), when both age and BP were adjusted for simultaneously. In conclusion, platelet activation was associated with arterial stiffness, suggesting that arterial stiffness may play an important role in thrombotic events.

Impact of shift work and race/ethnicity on the diurnal rhythm of blood pressure and catecholamines.

To evaluate the effects of shift work and race/ethnicity on the diurnal rhythm of blood pressure and urinary catecholamine excretion of healthy female nurses, 37 African American women and 62 women of other races underwent ambulatory blood pressure monitor and urine collection for 24 hours that included a full work shift: day shift (n=61), evening shift (n=11), and night shift (n=27). Awake and sleep times were evaluated from subjects' diaries. Of African Americans, 79% who were working evenings or nights and 32% working day shifts were nondippers (<10% drop in systolic pressure during sleep), whereas only 29% of others working evening+night and 8% working day shifts were nondippers. Regression analyses indicated that evening+night shift workers had a 5.4 mm Hg (P<0.001) smaller drop than day shift workers, and African Americans had a 4.0 mm Hg (P<0.01) smaller drop than others. The odds of an evening+night shift worker being a nondipper were 6.1 times that of a day shift worker (P<0.001), and the odds of an African American were 7.1 times that of others (P<0.001). Total sleep time was significantly greater in the non-African American day shift workers than in the other 3 groups. After controlling for work shift and race/ethnicity, we determined that longer sleep times predicted less dipping (absolute and relative) in blood pressure. Urinary norepinephrine and epinephrine were higher during work than nonwork in both racial groups of day shift workers, but in evening+night shift workers the difference was small and in the opposite direction. These results indicate that being African American and working evening or night shifts are independent predictors of nondipper status. Higher sleep blood pressure may contribute to the known adverse effects of shift work.

Direct visualization of copper-metallothionein in LEC rat kidneys: application of autofluorescence signal of copper-thiolate cluster.

We report on the histochemistry of copper-metallothionein (Cu-MT) in the kidneys of Long Evans Cinnamon (LEC) rats. We used the visualization principle of histochemistry based on the autofluorescence emission from the fluorophore of Cu(+)-thiolate clusters in proteins. Intense autofluorescence signals were observed with a ring at the outer stripe of the outer medulla. Orange fluorescence signals were observed in the nuclei and cytoplasm of proximal straight tubular (PST) cells of segment 3 (S3) at the outer stripe of the outer medulla, and yellow-orange signals were detected in lysosome-like organelles in the proximal convoluted tubule (PCT) cells of segments 1 and 2 (S1 and S2) adjacent to the glomeruli in the cortex. These fluorescent materials were identified as Cu-MT because both signals were quenched by withdrawing Cu+ or by blocking cysteine residues, the distributions of cysteine residues and immunoreactive MT showed identical patterns to the localization of the fluorescence signals, and the fluorescent proteins containing Cu were eluted at the same Kd value of purified Cu-MT by gel filtration chromatography. However, a high level of MT mRNA was detected only in the outer stripe of the outer medulla where the orange fluorescence signals were detected, but not in the cortex. This difference in localization between the protein and the mRNA suggested that synthesis of renal MT occurs do novo in the outer stripe of the outer medulla. The yellow-orange fluorescent Cu-MT located in the lysosomal organelles at S1 and S2 of the PCT cells in the cortex could be Cu-MT of nonrenal origin, i.e., Cu-MT transported from other organs.

Delayed recovery of postexercise blood pressure in patients with chronic heart failure.

Thirty-four patients with idiopathic dilated and ischemic cardiomyopathy underwent a symptom-limited cardiopulmonary exercise testing to evaluate the significance of postexercise blood pressure (BP) response. The postexercise BP response was useful in assessing the impaired exercise capacity and increased sympathetic activity in patients with heart failure.

Effect of diltiazem on sympathetic hyperactivity in patients with vasospastic angina as assessed by spectral analysis of arterial pressure and heart rate variability.

The autonomic nervous system importantly regulates coronary arterial tone and vascular resistance. To evaluate a role of autonomic nervous activity and the effects of calcium antagonist in patients with vasospastic angina (VSA), 13 VSA patients with patent coronary arteries (58+/-8 years) and 8 normal subjects (58+/-12 years) were studied. Arterial pressure and electrocardiogram were continuously recorded with the patient in a supine position under controlled respiration (0.2 Hz). Low-frequency (LF) and high-frequency (HF) components of the beat-to-beat variabilities of systolic arterial pressure and RR interval were then estimated by autoregressive power spectral analysis. The LF power (normalized unit) of both systolic arterial pressure (0.53+/-0.17 vs 0.30+/-0.17, p < 0.01) and RR variabilities (0.51+/-0.20 vs 0.31+/-0.16, p < 0.05) in patients with VSA were greater than that in normal subjects. There was no significant difference in the HF power. Seven patients with VSA who were treated with diltiazem (60 to 200 mg/day) had normalized LF power (normalized unit) of both systolic arterial pressure (0.62+/-0.12 vs 0.33+/-0.16, p < 0.01) and RR variabilities (0.55+/-0.23 vs 0.36+/-0.14, p < 0.05), together with clinical improvement. An increased sympathetic vasomotor tone and cardiac sympathetic predominance may play an important role in patients with VSA. Diltiazem improves these sympathetic hyperactivities.

Multinucleated variant endothelial cells (MVECs) of human aorta: expression of tumor suppressor gene p53 and relationship to atherosclerosis and aging.

Multinucleated variant endothelial cells (MVECs) generally exist in atherosclerotic human aorta and even in nonatherosclerotic aorta. Because the number of nuclei is increased in every MVEC, and because DNA instability was suspected, a series of oncogene expressions was conducted to clarify the nature of nuclear abnormality. The tumor suppressor gene p53 was found to be specifically expressed in the multinuclei of MVECs, while double nuclei were sometimes positive, and mononuclear typical endothelial cells were always negative for p53. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) revealed extra bands in exons 5 and 7 of the p53 gene, but no additional band in exons 6 and 8. In a BCL family, BCL-2 was coexpressed in one or two nuclei in the perinuclear space of the multinuclei of MVECs, whereas MCL-1, BCL-XS/L, and BAX were all negative, indicating that the BCL-2 coding gene is expressed only in the corresponding one or two nuclei of the multinuclei. Another oncogene, c-MET (hepatocyte growth factor receptor), was universally expressed in either type of endothelial cells, but other oncogenes, k-RAS and c-ERBB2, were not expressed in either type. MVECs were derived from human aorta and therefore non-tumorous somatic cells. No morphologic evidence of apoptosis was found. Although it is unclear that the extra bands came from the MVECs or just from ECs associated with atherosclerosis, combined immunocytological studies and PCR analysis suggest that MVECs express mutant type p53.

Probable sites of action of cyclic adenosine 3',5'-monophosphate in the induction of phosphodiesterase in Dictyostelium discoideum.

The sites of action cyclic adenosine 3',5'-monophosphate (cAMP) in phosphodiesterase [EC 3.1.4.17] induction in Dictyostelium discoideum were studied. When cAMP was added to the cell suspension from the start of the incubation, the effect of the cyclic nucleotide on the cellular enzyme-induction did not appear for 30 min, then occurred abruptly. From experiments on the addition of cAMP to the cell incubation mixture at various times, preparations for the synthesis of the enzyme appear to occur during the period from 20 min to 30 min after the start of the incubation. After the addition of cycloheximide at 30 min, the enzyme was degraded very rapidly. The half-life of phosphodiesterase was roughly 21 min in the presence of cAMP, and 12 min in its absence. The degradation rates became approximately the same on removing cAMP. When daunomycin and actinomycin D were added to cells previously stimulated with cAMP, phosphodiesterase was still synthesized at a higher rate than in cells not pretreated with cAMP. These results suggest that cAMP acts at two sites at least, i.e., on enzyme synthesis at the transcription level, and in suppressing the degradation of phosphodiesterase.

Participation of calcium in the induction of phosphodiesterase by cyclic adenosine 3',5'-monophosphate in Dictyostelium discoideum.

The effects of divalent cations on the induction of phosphodiesterase [EC 3.1.4.17] by cyclic adenosine 3',5'-monophosphate (cyclic AMP) were studied in Dictyostelium discoideum. When cells were incubated with 1 mM ethylene glycol-bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) in 20 mM Tris-HCl buffer, pH 7.5, for 2 h, the induction of cellular phosphodiesterase was inhibited by about 80%, and that of extracellular phosphodiesterase by about 65%. When cells were incubated with 1 mM EGTA for 1 h, 2 mM CaCl2 was added and the cells were further incubated for 1 h, the activities of cellular and extracellular phosphodiesterases were increased about 5 and 2.5 times, respectively, compared with those in the EGTA-inhibited cells. Although various other kinds of divalent cations were also studied, Ca2+ had the greatest effect on the induction. These results suggest that Ca2+ may participate in the induction of phosphodiesterase, and thus in the regulation of the development of the cellular slime mold.

Comparison of properties of the cellular and extracellular phosphodiesterases induced by cyclic adenosine 3',5'-monophosphate in Dictyostelium discoideum.

The kinetic properties and susceptibilities to various agents of intracellular (particulate and soluble) and extracellular phosphodiesterases [EC 3.1.4.17] of Dictyostelium discoideum induced by cyclic adenosine 3',5'-monophosphate (cyclic AMP) were studied and compared. Intracellular particulate phosphodiesterase was obtained by solubilization of the light mitochondrial fraction with Emulgen. The Michaelis constants of this enzyme were 4.5 +/- 0.7 and 10 +/- 0.7 microM, while those of the intracellular soluble phosphodiesterase were 4.6 +/- 0.3 and 13 +/- 2.8 microM. However, the Michaelis constant of the extracellular phosphodiesterase was 6.8 +/- 0.9 microM, differing from the values of the two intracellular enzymes. Susceptibilities of the enzyme activity to various agents (theophylline, caffeine, dithiothreitol, glutathione, etc.) were essentially the same among these three phosphodiesterases. In the presence of 10 mM ethylenediaminetetraacetate, the activities of the particulate and the soluble enzymes were both decreased to about 60%, while that of the extracellular enzyme remained at 90%. The inhibition constants of cyclic inosine monophosphate for the cellular enzymes (35 and 100 microM for the particulate enzyme, and 37 and 90 microM for the soluble one) were considerably different from the value for the extracellular enzyme (48 microM). These results suggest that the characteristics of these three phosphodiesterases are substantially similar, but that the affinity of the intracellular (particulate and soluble) enzymes for the substrate is somewhat different from that of the extracellular enzyme.

Intracellular localization of phosphodiesterase induced by cyclic adenosine 3',5'-monophosphate in Dictyostelium discoideum.

The intracellular distribution of phosphodiesterase [EC 3.1.4.17] induced by cyclic adenosine 3',5'-monophosphate (cAMP) in Dictyostelium discoideum was studied. When cAMP-treated cells were homogenized and fractionated according to the method of de Duve et al. ((1955) Biochem, J. 60, 604), the specific activity of phosphodiesterase was highest in the light mitochondrial fraction. Peaks of specific activities of alkaline phosphatase (marker enzyme of membrane) and catalase (marker enzyme of peroxisomes) also appeared in the same fraction as phosphodiesterase. However, after centrifugation of the light mitochondrial fraction in a sucrose density gradient, the activity of phosphodiesterase was clearly separated with that of catalase (density 1.19 g/ml) and showed three peaks at lower density (1.10, 1.13, 1.17 g/ml) with good reproducibility. Some parts (1.13, 1.17 g/ml) of the activity in the gradient overlapped with alkaline phosphatase activity, but in the density fraction of 1.10 g/ml the activity of alkaline phosphatase was hardly detectable. When the light mitochondrial fraction was treated with Emulgen 108, or sonicated, phosphodiesterase was more easily solubilized than alkaline phosphatase and catalase, and was found in supernate after centrifugation at 20,000 X g for 30 min. In order to distinguish the locations of the three enzymes, the supernatant of the light mitochondrial fraction treated with Emulgen 108 was subjected to charge shift electrophoresis. The electrophoretic mobilities of phosphodiesterase and catalase were unaffected by ionic detergent. However, alkaline phosphatase shifted towards the anode in the presence of anionic detergent (sodium deoxycholate), and shifted towards the cathode in cationic detergent (cetyltrimethylammonium bromide), relative to nonionic detergent (Emulgen 108) alone. Thus, some part of the phosphodiesterase induced by cAMP may be associated with the plasma membrane, but the remainder is localized in some kind of intracellular particle of lower density. Moreover, the association with the membrane or particle is more easily dissociated than that of alkaline phosphatase, and the liberated phosphodiesterase is rather hydrophilic.

Association of the liver peroxisomal fatty acyl-CoA beta-oxidation system with the synthesis of bile acids.

The association of liver peroxisomal fatty acyl-CoA beta-oxidizing system (FAOS) with the synthesis of bile acids was investigated. When rats were given clofibrate, a peroxisome proliferator and stimulator of peroxisomal FAOS, the biosynthesis of bile acids was significantly increased. Di(2-ethylhexyl)phthalate, another peroxisome proliferator, also increased the biosynthesis of bile acids. On the other hand, administration of orotate, an inhibitor of mitochondrial FAOS activity, did not affect the biosynthesis. It is known that fatty acyl-CoA oxidase [EC 1.3.99.3] in peroxisomal FAOS conjugates with catalase [EC 1.11.1.6]. When the catalase activity of liver peroxisomes was irreversibly inhibited by administration of 3-amino-1,2,4-triazole (amino-triazole), the biosynthesis of bile acids was suppressed to about one-third, and the serum cholesterol level was increased. However, the bile acid components of the bile obtained from aminotriazole-treated rats were not essentially different from those of control rats, and no accumulation of intermediates of bile acid synthesis was found in this experiment. Peroxisomal FAOS activity of the liver from amino-triazole-treated rats was considerably lower than that of control liver. The above results indicate that liver peroxisomes play a role in the biosynthesis of bile acids in vivo.

The origin of lipid accumulated in liver lysosomes after administration of triton WR-1339.

The origin of the lipids accumulated in liver lysosomes after administration of Triton WR-1339 was investigated. When Triton WR-1339 was injected into rats, serum triglyceride and cholesterol increased markedly. The highest content of triglyceride was observed in the second-day serum, from which very-low-density lipoprotein (VLDL) was isolated. The VLDL was administered to normal rats, then the light mitochondrial fraction of the liver at 24 h was centrifuged in a sucrose density gradient. The activities of lysosomal enzymes, acid phosphatase, N-acetyl-beta-D-glucosaminidase and acid lipase, were all shifted to less dense fractions as compared with those of normal lysosomes. [3H]Triglyceride-labeled VLDL was injected similarly, and at 12 and 24 h after the administration, the light mitochondrial fraction of the liver was fractionated by sucrose gradient centrifugation. Protein content and radioactivity in the immunoprecipitate with anti-VLDL serum at 12 h showed almost the same distribution as acid phosphatase activity. At 24 h, though acid phosphatase activity, immunoprecipitable protein content and radioactivity were all found in less dense fractions than in the case of normal lysosomes, the former two distributions were significantly different from the latter. The anti-VLDL serum reacted in Ouchterlony tests not only with Triton-induced VLDL and normal VLDL but also with the extract from low-density lysosomes. These results suggest that the lipids accumulated in low-density lysosomes following the administration of Triton WR-1339 were probably derived from the elevated serum VLDL induced by the treatment.

Expression of human metallothionein-2 in Escherichia coli: cadmium tolerance of transformed cells.

A genetic approach was undertaken to investigate the physiological roles of human metallothionein-2. A constructed expression plasmid, pEXPMTII, in which human metallothionein-IIA cDNA was inserted downstream of a tryptophan-lactose promoter, was used to transform Escherichia coli JM105 strain. Cadmium-binding metallothionein was successfully expressed in E. coli in the medium containing cadmium, while copper and zinc-metallothioneins were scarcely observed in copper- or zinc-containing medium. The amino acid composition and sequence of the biosynthesized cadmium-metallothionein were analyzed. The selectivity of metals bound to metallothionein and the stability of metal-binding forms of metallothionein in E. coli were discussed. In addition, cadmium, zinc, or copper resistance of the cells expressing metallothionein was examined. Cells transformed with the plasmid pEXPMTII and cultured in a medium containing cadmium exhibited tolerance only to cadmium. It was demonstrated that human metallothionein-2 functioned for cadmium detoxification in E. coli.

Apoptosis and Ki-67 expression in adenomyotic lesions and in the corresponding eutopic endometrium.

OBJECTIVE: To examine biologic and proliferative properties of adenomyotic lesions and to determine whether adenomyotic lesions originate in the basal layer of the eutopic endometrium. METHODS: We examined eutopic and ectopic endometria from 23 patients with adenomyosis. To obtain evidence for the induction of programmed cell death, apoptotic cells were identified using a modified terminal deoxynucleotidyltransferase-biotin nick end-labeling method. To evaluate cell death repressor activity, bcl-2 gene expression was examined using immunohistochemical staining. As a proliferative marker, Ki-67 expression was also examined immunohistochemically. RESULTS: In the eutopic endometrium, apoptosis was most frequently observed in epithelial cells during mid- to late secretory phases, although it was rarely found during early proliferative through early secretory phases (P<.01). In contrast, bcl-2 gene expression inversely correlated with the appearance of apoptosis. A similar tendency was observed in stromal cells. In the ectopic endometrium of adenomyosis, endometrial dating revealed that secretory change was rare, even in the secretory phase, and that induction of apoptotic cells as well as bcl-2 gene expression showed no cyclic change. In stromal cells of the ectopic endometrium, apoptosis was more frequent than was seen in the eutopic endometrium, in all menstrual phases (P<.05). Ki-67 was constantly expressed in the glandular epithelium of the ectopic endometrium, irrespective of the menstrual phases, whereas in the secretory phase it was less expressed in the eutopic endometrium of functional and basal layers (P<.01). CONCLUSION: The induction of apoptosis seems to be regulated by hormonal changes in the eutopic endometrium and has an inverse correlation with bcl-2 gene expression. The ectopic endometrium in adenomyosis is rarely influenced by hormonal change and has different biologic and proliferative properties than events observed in the eutopic endometrium findings, which strongly suggest that the adenomyotic lesion does not originate in the basal endometrium.

Loop and distal actions of a novel diuretic, M17055.

We investigated the mechanism of action of a novel 'high ceiling' diuretic, M17055, in in vivo clearance studies with anesthetized dogs during water diuresis and in vitro microperfusion studies of isolated rabbit renal tubules. In the clearance study, intravenous infusion of M17055 (1 mg/kg per h) decreased free water clearance and increased urinary excretion of Na+ and Cl- to a greater extent than did a maximum dose of furosemide (30 mg/kg per h). With the maximum dose of furosemide, an additional dose of M17055 or hydrochlorothiazide resulted in additional suppression of free water clearance. These results indicate that M17055 has some additional mechanisms of action in the distal nephron. In isolated rabbit cortical thick ascending limb of Henle's loop, M17055 applied to the lumen decreased the lumen positive transepithelial voltage at concentrations over 10(-6) M and suppressed the lumen-to-bath 36Cl- flux at 10(-5) M. In the connecting tubule, M17055 added to the lumen suppressed lumen negative transepithelial voltage in a concentration-dependent manner in a range from 10(-4) to 10(-3) M. The effect of M17055 on transepithelial voltage was also observed in the distal convoluted tubule and cortical collecting duct. Moreover, 10(-3) M of M17055 in the lumen significantly decreased the lumen-to-bath 22Na+ flux in the cortical collecting duct. From these observations, it appears that M17055 acts not only on the thick ascending limb of Henle's loop but also on the distal segments via inhibition of electrogenic Na+ transport.