Common occurrence of everolimus-associated aphthous stomatitis in Japanese heart transplant recipients.

Mammalian target of rapamycin (mTOR) inhibitors display antiproliferative effects with less nephrotoxicity than calcineurin inhibitors. However, clinical use of mTOR inhibitors can be associated with a series of adverse events. We experienced cases of aphthous stomatitis associated with everolimus (EVL) in four Japanese heart transplant recipients treated at the target trough EVL blood level after a switch from mycophenolate mofetil between April and December 2007. All four patients developed aphthous stomatitis; three required reduction of the exposure and one, EVL discontinuation due to stomatitis as well as other side effects. All patients recovered from stomatitis after reduction or withdrawal of EVL. Thus, we considered that EVL-related stomatitis might occur commonly among the Japanese population. The proper dosage, effects, and frequency of the side effects of mTOR inhibitors may vary by ethnic population.

Heterogeneity of regional systolic function detected by tissue Doppler imaging is linked to impaired global left ventricular relaxation in hypertrophic cardiomyopathy.

OBJECTIVE: To evaluate regional and global left ventricular (LV) function and LV wall thickness (LVWT) in patients with hypertrophic cardiomyopathy (HCM). DESIGN AND SETTING: Observational study at the National Cardiovascular Centre and Nagoya University Hospital in Japan. PARTICIPANTS: Thirty-six patients with HCM and 16 patients with hypertensive LV hypertrophy (LVH). MAIN OUTCOME MEASURES: Conventional echocardiography and strain rate (SR) imaging derived from tissue Doppler imaging were performed. Systolic strain (epsilon(sys)), peak systolic SR (SR(sys)), peak early diastolic SR (SR(dia)) and LVWT were obtained from eight LV segments. LV pressure was simultaneously recorded with a high-fidelity micromanometer. RESULTS: The regional epsilon(sys) and SR(sys) were correlated with LVWT in patients with HCM (r = 0.50, p<0.001 and r = 0.63, p<0.001, respectively) but not in patients with hypertensive LVH. The standard deviations of LVWT, epsilon(sys) and SR(sys) obtained from the eight LV segments of each subject were greater for patients with HCM than for patients with hypertensive LVH. The standard deviation of LVWT was correlated with those of epsilon(sys) and SR(sys) (r = 0.55, p<0.001 and r = 0.56, p<0.001, respectively). The standard deviations of LVWT, epsilon(sys) and SR(sys) were correlated with tau (r = 0.35, p<0.05; r = 0.47, p<0.001; and r = 0.39, p<0.005, respectively). CONCLUSIONS: Heterogeneity of regional LV systolic function detected by SR imaging is in part attributable to heterogeneity of LVH and may be linked to impaired global LV relaxation in HCM.

Heart-type fatty acid binding protein is a novel prognostic marker in patients with non-ischaemic dilated cardiomyopathy.

OBJECTIVE: To determine whether concentrations of heart-type fatty acid binding protein (H-FABP) measured before hospital discharge predict critical cardiac events in patients with idiopathic dilated cardiomyopathy (DCM). PATIENTS: 92 consecutive patients with DCM were enrolled and followed up for four years. MAIN OUTCOME MEASURES: Serum concentrations of H-FABP, brain natriuretic peptide (BNP), cardiac troponin T before hospital discharge and survival rate. RESULTS: 23 patients died of cardiac causes, received a left ventricular assist device or underwent heart transplantation during the four-year follow up. Univariate analyses showed that New York Heart Association functional class, heart rate, ejection fraction, serum H-FABP and plasma BNP were significant variables. According to multivariate analysis, serum H-FABP and plasma BNP concentrations were independent predictors of critical cardiac events. Cardiac troponin T before hospital discharge was not a predictor. The area under the receiver operating characteristic curve for death from critical cardiac events was similar between H-FABP and BNP. Patients with an H-FABP concentration at or above the median (> or = 5.4 ng/ml) had a significantly lower survival rate than those below the median, according to analysis by log rank test (p < 0.0001). When combined with BNP concentration at or above the median (> or = 138 pg/ml), H-FABP below the median predicted the worst prognosis among the combinations. CONCLUSIONS: The concentration of serum H-FABP before discharge from hospital may be an independent predictor for critical cardiac events in DCM.

Mechanical strain stimulates a mitogenic response in coronary vascular smooth muscle cells via release of basic fibroblast growth factor.

Mechanical strain has been shown to induce mitogenesis in a rat neonatal vascular smooth muscle (VSM) cell line in a response mediated predominantly by transcription, expression, and release of platelet-derived growth factor (PDGF). We examined the effect of cyclic mechanical strain and growth factor production on mitogenic response in ovine coronary artery smooth muscle cells. Vascular smooth muscle cells were cultured from explants of left anterior descending (LAD) coronary arteries from young sheep. Cells for experiments were grown on wells with silicone-elastomer bottoms, and subjected to strain (60 cycles/min) using a vacuum actuated strain device. Tritiated thymidine incorporation was used as a measure of DNA synthesis. Cell membrane damage was assessed with differentially permeable nuclear staining dyes. We observed an increase in tritiated thymidine incorporation in response to strain with a temporal response identical to that observed in response to exogenous growth factors (PDGF-BB and basic fibroblast growth factor [bFGF]). Supernatant medium obtained from stretched cells induced a twofold increase in DNA synthesis in unstretched cells. The mitogenic response was abolished by monoclonal antibodies to bFGF, but not by antibodies to PDGF-AB. Studies of fluorescent dye exclusion indicated the stretching protocol caused no cell membrane damage. Thus, mechanical strain is an important stimulus for growth factor release in coronary VSM cells. The mitogenic response is mediated by release of bFGF.

Structure-activity study and analgesic efficacy of amino acid derivatives as N-type calcium channel blockers.

The synthesis and structure-activity relationship (SAR) study of a novel series of N-type calcium channel blockers are described. L-Cysteine derivative 2a was found to be a potent and selective N-type calcium channel blocker with IC(50) 0.63 microM on IMR-32 assay. Compound 2a showed analgesic efficacy in the rat formalin-induced pain model by intrathecal and oral administration.

Nicorandil, a hybrid between nitrate and ATP-sensitive potassium channel opener, preconditions human heart to ischemia during percutaneous transluminal coronary angioplasty.

The human heart progressively becomes more tolerant to ischemia after repeated balloon inflations during percutaneous transluminal coronary angioplasty (PTCA). The present study investigated whether nicorandil, a hybrid between nitrate and an ATP-sensitive potassium channel opener, affects this ischemic preconditioning. Sixteen patients with stable angina pectoris caused by left anterior descending artery lesions were subjected to 2 balloon inflations of 2-min duration with a 3-min reperfusion period. Seven of these patients served as the control group and in the remaining 9 patients, nicorandil was administered intravenously (6 mg/h) throughout the PTCA procedure (nicorandil group). The lactate extraction ratio (LER) was obtained at 30 s after each ischemic event (LERpost-1 and LERpost-2) in both groups. In the control group, LERpost-1 was more negative than LERpost-2 (-185.7+/-74.2 vs -98.0+/-37.3%, p<0.01). The ratio of the sum of the ST elevation in the precordial leads during the second inflation (sumST-2, 0.94+/-0.66 mV) to that during the first inflation (sumST-1, 1.43+/-1.17 mV) was 0.72+/-0.16 in the control group, which was less than the ratio in the nicorandil group (1.06+/-0.13, p<0.01). Nicorandil abolished the difference between the 2 ischemic events (LERpost-1, -45.1+/-41.6 vs LERpost-2, -43.5+/-51.1%; sumST-1, 1.38+/-0.80 vs sumST-2, 1.46+/-0.90 mV). LER was less negative in the nicorandil group than that in the control group (LERpost-1, -45.1+/-41.6 vs -185.7+/-74.2%, p<0.01; LERpost-2, -43.5+/-51.1 vs -98.0+/-37.3%, p<0.05). Thus, nicorandil improved lactate metabolism during PTCA without significantly influencing ST-elevation. In conclusion, intravenous pre-administration of nicorandil appears to precondition the human heart during PTCA.

Transcardiac 8-iso-prostaglandin F(2 alpha)generation from acute myocardial infarction heart: insight into abrupt reperfusion and oxidant stress.

8-iso-prostaglandin F(2 alpha)(8-iso-PGF(2 alpha)), a representative isoprostane, has been reported to be a reliable marker for oxidant stress in vivo. To examine if 8-iso-PGF(2 alpha)is generated in patients with acute myocardial infarction (AMI), we measured the level of immunoreactive 8-iso PGF(2 alpha)in the great cardiac vein as well as classical eicosanoids, 6-keto-prostaglandin F(1 alpha)(6-keto-PGF(1 alpha)) and thromboxane B(2)(TXB(2)) in the process of urgent coronary balloon angioplasty. Fourteen patients with anterior AMI were divided into two groups: the totally occluded (n=7) and the already perfused groups (n=7). In the former, transient elevation of 8-iso-PGF(2 alpha)was observed immediately after the angioplasty, i.e. the ratio of post-angioplasty level to pre-level was approximately 2.4 for 8-iso-PGF(2 alpha), 14 for 6-keto-PGF(1 alpha), and 5 for TXB(2). In the already perfused group, the levels of these eicosanoids were unchanged. In the totally occluded group, peak creatine phosphokinase in a peripheral vein was correlated with the level of 8-iso-PGF(2 alpha)(r(2)=0.841, P<0.01), but not with those of the other two eicosanoids. In conclusion, transcardiac 8-iso-PGF(2 alpha)generation is a reliable marker for the size of myocardium exposed to oxidant stress.

Impact of coronary risk factors on contribution of nitric oxide and adenosine to metabolic coronary vasodilation in humans.

OBJECTIVES: The contribution of nitric oxide (NO) and adenosine to the increase in coronary blood flow (CBF) induced by cardiac pacing was investigated in 28 subjects with angiographically normal coronary arteries with and without one or more risk factors for atherosclerosis. BACKGROUND: NO and adenosine are important in the regulation of coronary circulation, and the inhibition of NO synthesis increases adenosine production during cardiac pacing in experimental models. METHODS: Coronary artery diameters and CBF were assessed by quantitative coronary arteriography and Doppler flow velocity measurement. Plasma levels of nitrites and nitrates (NOx) (stable end products of NO), adenosine and lactate were measured, and blood gas analysis was performed. RESULTS: The extent of CBF response to cardiac pacing did not differ between the 14 subjects with and the 8 subjects without risk factors for atherosclerosis. NOx (12.0+/-0.9 vs. 14.9+/-1.1 ,amol/liter [mean+/-SD], p < 0.05), but not adenosine (50.8+/-7.2 vs. 50.8+/-6.5 nmol/liter), levels in coronary sinus blood increased in the subjects without risk factors. In contrast, adenosine (58.9+/-7.5 vs. 77.4+/-9.8 nmol/liter, p < 0.05), but not NOx (11.1+/-1.1 vs. 12.2+/-1.1 micromol/liter), levels increased in subjects with risk factors. Aminophylline, an antagonist of adenosine receptors, blunted CBF response to cardiac pacing in six subjects with risk factors. The number of risk factors showed a negative correlation (p < 0.05) with NOx production and a positive correlation (p < 0.05) with adenosine production during cardiac pacing, respectively. CONCLUSIONS: NO and adenosine are increased during metabolic coronary vasodilation induced by cardiac pacing. Adenosine production may be a compensatory mechanism when NO production is reduced.

Intravascular ultrasonic evidence by constant cross-sectional area of atherosclerotic plaques during coronary vasomotion in humans.

AIMS: This study aims to visualize ultrasonically deformation of atherosclerotic plaques in human coronary arteries during vasoconstriction and vasodilatation. METHODS AND RESULTS: Intravascular ultrasound detected occult atherosclerosis in angiographically normal coronary arteries of eight patients with chest pain at rest. During the acetylcholine provocative test, intravascular ultrasound monitored deformation of the atherosclerotic plaques. At the last step of the test, intracoronary injection of isosorbide dinitrate caused vasodilation. Under control, acetylcholine-treated, and isosorbide dinitrate-treated conditions, cross-sectional areas of sonolucent circle and vessel lumen were measured. Subtraction of the latter from the former gave the area of atherosclerotic plaque. In the process of vasoconstriction and vasodilation, the plaque area did not change significantly. CONCLUSION: The cross-sectional area of the atherosclerotic plaque appeared to be constant during vasomotion of human coronary arteries.

Effect of theophylline on adaptation of the heart to myocardial ischemia during percutaneous transluminal coronary angioplasty in patients with stable angina pectoris.

This study was designed to examine whether theophylline, an adenosine receptor antagonist, affects cardiac adaptation to ischemia during progression of repetitive balloon inflations of percutaneous transluminal coronary angioplasty (PTCA). Theophylline abolished this cardiac adaptation, suggesting that endogenous adenosine is a key mediator for cardiac adaptation during PTCA.

Growth pattern of experimental squamous cell carcinoma in rat submandibular glands--an immunohistochemical evaluation.

Histopathological and immunohistochemical studies during carcinogenesis in rat submandibular glands (SMGs) using a carcinogen (9,10-dimethyl-1,2-benzanthracene: DMBA) were evaluated. For carcinogenesis, the carcinogen-containing sponge was surgically inserted into the gland. Histopathological features during carcinogenesis were as follows; dilatation of ductal segments, the presence of duct-like structures and cystic lesion around the sponge were observed within 3 weeks of the experiment, squamous metaplasia in duct-like structures and lining epithelium of the cystic structures around the sponge were observed at 4-6 weeks of the experiment, and finally well differentiated squamous cell carcinomas (SCCs) were observed after 8 weeks of the experiment. The immunoreactivity of K8.12 keration (K8.12), S-100 protein (S-100), epidermal growth factor (EGF), laminin, and proliferating cell nuclear antigen (PCNA) were evaluated. In the normal SMG, EGF was confined to the granular cells and S-100 to the pillar cells of granular convoluted tubules (GCTs). K8.12 was found in striated (SD) and excretory duct (ED) cells and laminin showed linear staining of the basement membrane around the ducts, acini and blood vessels. PCNA-positive nuclei were rarely observed in the normal glandular parenchyma. During carcinogenesis, during the first stage, EGF in granular cells and S-100 in pillar cells of GCT segments disappeared, and cytokeration K8.12 was observed in duct-like structures and cystic epithelium around the DMBA sponge. PCNA-positive nuclei in the first stage were mainly confined to basal cells of morphologically altered ducts. During the second stage, squamous metaplastic cells showed an intense K8.12 reaction. During the third stage, the well differentiated SCC showed strong reaction for K8.12, and the linear staining for laminin staining had disappeared at the invading fronts. The PCNA index was nearly 40% in the tumour cell component. The stem cells or the progenitor cells during experimental carcinoma were most likely to be the ductal basal cells, and carcinogenesis was initiated with an increase of proliferating activity in small cell clusters surrounding a necrotic area, basal cells of dilated excretory ducts and duct-like structures. Thus, all ductal segments undergoing squamous metaplasia may participate in the genesis of neoplasia during experimental carcinogenesis.

Comparison of proliferating cell nuclear antigen index in benign and malignant salivary pleomorphic adenoma.

The expression of proliferating cell nuclear antigen (PCNA) was studied in benign and malignant pleomorphic adenomas by using monoclonal antibody to PCNA. Carcinoma in pleomorphic adenoma (n = 8), cell-rich variant (n = 6) and typical pleomorphic adenoma (n = 6) were selected in this study. The PCNA index in carcinoma in pleomorphic adenoma showed a higher index of nuclear staining (mean 22.9%, S.D. 6.2) than in typical pleomorphic adenoma (mean 6.9%, S.D. 3.4) or a cell-rich variant of pleomorphic adenoma (mean 8.8%, S.D. 3.3). A significant difference in PCNA index was found between benign and malignant pleomorphic adenoma (P < 0.05). The present study suggests that PCNA index significantly differs between pleomorphic adenoma and carcinoma in pleomorphic adenoma, but in the prediction of malignant transformation potential it should be combined with routine histopathological examination.

Immunohistochemical detection of S-100, S-100 alpha, S-100 beta proteins, glial fibrillary acidic protein, and neuron specific enolase in the prenatal and adult human salivary glands.

Developing human fetal salivary glands of gestational age from 10 to 40 weeks (n = 100) and normal adult glands (n = 10) were examined for immunoreactivity to S-100 protein and its subunits S-100 alpha, S-100 beta, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE). In the early intermediate developmental stage (19-32 weeks) some acinar basal cells showed immunoreactivity to S-100 protein which rapidly disappeared in the late developmental stage (33-40 weeks). Adult salivary glands were negative for S-100 protein. The S-100 alpha subunit was strongly positive in the glandular ducts and acini of both fetal and adult glands. The S-100 beta, although present in some acini and ductal cells during the late intermediate developmental stage, was rarely seen in the adult glands. GFAP and NSE was positive at the developing salivary epithelium in the early developmental stage (15-18 weeks). The above findings indicated that the developing salivary epithelia showed transient appearance of the neuronal phenotype during active cytodifferentiation stage of glandular acini and ducts. Therefore, after evaluation of normal developmental and neoplastic transformation of the salivary glands a suggestion that neuronal differentiation of ductal reserve cells is responsible for the production of modified myoepithelial cells in both normal developmental salivary gland and neoplastic transformation is made.

Immunoreactivity of proliferating cell nuclear antigen in salivary gland tumours: an assessment of growth potential.

Immunoreactivity of proliferating cell nuclear antigen (PCNA) was assessed to evaluate growth potential in surgically resected tissue specimens from 70 cases of benign and malignant salivary gland tumours. Three stage streptavidin-biotin immunoperoxidase immunostaining using monoclonal antibody to PCNA showed a heterogeneity of PCNA index and distribution. In normal salivary gland specimens, PCNA was demonstrated in the nuclei of few ductal and acinar cells. In pleomorphic adenoma a multiple nodular growth pattern was observed with positive immunoreactivity restricted to the nuclei of tubulo-ductal structures. Warthin's tumour had positive nuclei in the outer cuboidal cells of epithelial component and germinal centres of lymphoid tissue. Myoepithelioma and acinic cell carcinoma showed slightly differing values and a statistically significant difference in the value of the index was observed in tumour cell aggregates of the cribiform type of adenoid cystic carcinoma and the solid undifferentiated type and between low/intermediate and high-grade mucoepidermoid tumours. PCNA is a useful marker of tumour cell proliferation; the index correlates with the grade of malignancy in salivary gland tumours.

Immunohistochemical study of lymphoid tissue in human fetal salivary gland.

Immunoreactivity of lysozyme (LY), lactoferrin (LF), alpha 1-antichymotrypsin (alpha 1-ACT), alpha 1-antitrypsin (alpha 1-AT), keratin proteins KL1, PKK1, K8.12, S-100 protein, MAM-3, MAM-6, and epithelial membrane antigen (EMA) were evaluated in lymphoid and glandular tissues of developing salivary gland of human fetus (gestational age ranging from 17 to 40 wk to investigate the role of lymphoid tissue in developing salivary glands. In a total of 79 cases, lymphoid cell aggregations were noted in parotid (57 cases), submandibular (21 cases) and sublingual (5 cases) glands. Mononuclear cells showing intense activity of LY, alpha 1-ACT and alpha 1-AT were present in the lymphoid aggregation. The glandular ducts embedded in lymphoid tissue were negative to MAM-3, MAM-6, EMA and S-100 protein, but showed positive PKK1 and KL1 reaction during early stages of development, and showed degeneration and effacement upon increase in number and LY activity of the mononuclear cells. The lymphoid aggregations progressively emerged as lymph nodes.

Immunohistochemical and electronmicroscopic studies of obstructive lesions in submandibular glands.

Obstructive sialoadenitis was examined by immunohistochemical techniques for keratin (MoAb KL1, PKK1 and K8.12) and actin. Electronmicroscopy (EMS) was used to identify ultrastructural changes in myoepithelial cells and ductal basal cells. With immunohistochemistry, actin staining was used as a marker of myoepithelium, MoAbs KL1 and PKK1 for ductal luminal cells, and MoAb K8.12 for ductal basal cells. Histologic features of the lesion usually showed degenerative changes of acinar and duct cells with cell infiltration and fibrous replacement. Immunohistochemical findings indicated that actin staining in the changed myoepithelial cells was irregularly positive or negative, and also keratin staining in luminal and ductal basal cells was reduced or disappeared. Ultra-structural features of the changed myoepithelial cells indicated that these cells appeared less altered than adjacent acinar and ductal cells and showed increased amounts of lipid droplets and lipofuscin granules, and also wrinkled processes filled the prominent myofilament material.

Heterogeneity and multiple expression of intermediate filament proteins, S-100 protein and neuron specific enolase in skin mixed tumor.

Intermediate filament proteins, keratin (KL1, PKK1, K8.12) and vimentin, S-100 protein alpha and beta subunits and neuron specific enolase were evaluated immunohistochemically to determine their distribution patterns in the tumor components of mixed tumor of skin. Keratin proteins were distributed widely in tumor epithelial cells or modified myoepithelial (MME) or neoplastic myoepithelial (NME) cells. Luminal cells of the tubulo-ductal structure of the tumor mass showed positive staining of KL1 and PKK1 keratins and an infrequently positive reaction of MoAb K8.12. The outer or basal tumor cells were characterized by coexpression of K8.12 keratin, vimentin, S-100 protein and infrequently neuron specific enolase reactivity. Heterogeneity of keratin distribution was seen in tumor epithelial cells. MME cells or NME cells of skin mixed tumor showed coexpression of keratin and vimentin, and multiple expression of intermediate filament proteins, S-100 protein and neuron specific enolase. Hyaline and chondroid changed cells stained intensely to vimentin and S-100 proteins, as well as to neuron specific enolase. The authors evaluate the histogenesis of skin mixed tumor in relation to epithelial and myoepithelial cells of the sweat gland and their immunohistochemical findings.

Vimentin expression in sweat gland tumours.

Immunohistochemical localization of vimentin was studied in 93 cases of sweat gland tumours using a monoclonal anti-vimentin antibody. A strong immunoreactivity of vimentin was observed in modified myoepithelial or neoplastic myoepithelial cells of mixed tumour of the skin, syringoma, and sweat gland adenoma. Tumour cells in outer layers of tubular, ductal, and duct-like structures usually showed positive staining for vimentin, which coincided with modified myoepithelial cells. All tumour cells of clear cell hydroadenoma showed positive vimentin staining. Tumour cells of the luminal border of tubulo-ductal structures of mixed tumours were rarely immunoreactive for vimentin. Positive vimentin staining of tumour cells in the outer zone of tubulo-ductal structures in sweat gland tumours may be related to reactive proliferation of modified myoepithelial cells and simultaneous growth of luminal tumour cells.

Heterogeneity of keratin expression and actin distribution in benign and malignant mammary diseases.

Immunoreactivity of monoclonal anti-cytokeratin KL1, PKK1, K8.12 and anti-actin antibodies in 101 cases of diseased human breast lesions showed irregular keratin distribution in luminal cells of terminal ductal-lobular unit and basal layer cells of the interlobular and main duct. Actin staining was confined to myoepithelial cells. Benign lesions showed great heterogeneity in luminal cells of the terminal ductal-lobular units. Breast carcinoma showed a reduced staining for keratins, heterogeneity of keratin expression was found in solid tubular carcinoma, and actin was usually absent: however, papillo-ductal or comedo type had actin positive myoepithelial cells around carcinoma foci.