Esophageal adenosquamous carcinoma in a cat.

Esophageal carcinoma was observed in an eight-year-old, castrated male, Japanese domestic cat. Histologically, this neoplasm consisted of two different growth patterns, squamous cell carcinoma and adenocarcinoma. The results of immunohistochemical examination supported the fact that the two kinds of neoplastic cells have different characteristics. The tumor was, therefore, diagnosed as adenosquamous carcinoma. Esophageal tumors in the cat are very rare and, if any, neither adenocarcinoma nor adenosquamous carcinoma has been reported up to the present.

Expression of cytochrome P450 (CYP) isozymes in rat placenta through pregnancy.

The placenta plays a vital role in maintenance of pregnancy and is able to metabolize many foreign chemical compounds by cytochrome P450 (CYP) system. It is very important to clarify the expression of CYPs in the rat placenta, because rats are frequently and widely used in the embryo- and feto-toxicity studies on foreign chemical compounds. In this study, we investigated the changes in the expression and localization of CYPs protein in the placenta of F344 rats at 9, 11, 13, 16 and 19 days of gestation by Western blot analysis and immunohistochemical staining. Among nine isozymes examined (CYP1A1, 2B1, 2C6, 2C12, 2D1, 2D4, 2E1, 3A1 and 4A1), only CYP3A1 was clearly detected by Western blot analysis at all days of gestation examined. Immunohistochemically, the cytoplasm of giant cells in the trophoblastic region was positively stained by anti-CYP3A1 antibodies. These results suggest that CYP3A1 may be a major component of CYP system in the rat placenta.

Viability of Q-wave infarcted myocardium with restored positive and persistent negative T waves after optimal revascularization compared with dobutamine stress echocardiography.

To clarify the clinical difference in viability of myocardium with negative and positive T waves in Q-wave anterior or anteroseptal myocardial infarction, we performed low-dose dobutamine stress echocardiography in 17 patients with negative T waves and in 13 patients with positive T waves with optimal revascularization of infarct-related arteries in the chronic phase of infarction. At baseline the wall motion score (WMS) of the negative and positive T groups was 25.8 +/- 3.0 and 22.3 +/- 2.2 points (p <0.05), respectively. At peak stress WMS in each group was 27.2 +/- 4.2 and 19.8 +/- 2.4 points (p <0.0001), respectively. With dobutamine stress WMS in the positive T group was more decreased than that of the negative T group (p <0.0001). We conclude that the restored positive T waves in Q-wave myocardial infarction indicate a significantly greater amount of viable myocardium than the negative T waves, showing better regional wall motion improvement with low-dose dobutamine stress.

[Attenuation of reperfusion phenomenon by reperfusion using leukocyte-depleted blood during direct percutaneous transluminal coronary angioplasty for acute myocardial infarction].

Leukocytes are important in the occurrence of reperfusion injury in coronary intervention for acute myocardial infarction (AMI). This study compared reperfusion injury caused by reperfusion using leukocyte-depleted blood (LD) and that by conventional angioplasty (control) through the reperfusion phenomenon including reperfusion arrhythmia and additional ST elevation during direct percutaneous transluminal coronary angioplasty (PTCA) in 41 patients with 21 left anterior descending artery (LAD) lesions and 20 right coronary artery (RCA) lesions. LD was prepared from 20 ml of venous blood, 20 ml of mixed blood and 60 ml of arterial blood from the patients (LD group; LAD-LD: n = 10, RCA-LD: n = 10) which was passed through a leukocyte removal filter. The blood was injected from the tip of the balloon catheter at 10 ml/min during inflation for 10 min before balloon deflation. The control group (LAD-control: n = 11, RCA-control: n = 10) underwent conventional angioplasty. The appearance of reperfusion arrhythmia [atrioventricular block (AVB) > II, accelerated idioventricular rhythm (AIVR), ventricular tachycardia (VT), ventricular fibrillation (Vf)] and measurements of STmax deviation before and after reperfusion, the differences of the STmax deviation (delta ST) and additional ST elevation (LAD: > or = 0.5 mV increase of sigma ST in lead V1-V6, RCA: > or = 0.3 mV increase of sigma ST in lead II, III and aVF) were studied. The appearance of reperfusion arrhythmias was as follows; LAD-LD: AVB 0, AIVR 1, VT 1, Vf 0, LAD-control: AVB 0, AIVR 4, VT 2, Vf 1, NS, and RCA-LD: AVB 0, AIVR 0, VT 0, Vf 0, RCA-control: AVB 2, AIVR 0, VT 1, Vf 0, NS. There was no reperfusion arrhythmia in the RCA-LD group. There was no significant difference in the appearance of reperfusion arrhythmias between the LAD-LD and LAD-control or RCA-LD and RCA-control groups. Before reperfusion the STmax deviation (mV) was LAD-LD 0.86 +/- 0.46 vs LAD-control 0.74 +/- 0.49 and RCA-LD 0.29 +/- 0.18 vs RCA-control 0.15 +/- 0.09 and after reperfusion LAD-LD 0.63 +/- 0.35 vs LAD-control 0.81 +/- 0.49 and RCA-LD 0.13 +/- 0.15 vs RCA-control 0.20 +/- 0.12, respectively. There were no significant differences between LAD-LD and LAD-control or RCA-LD and RCA-control groups. delta ST (mV) was LAD-LD 0.23 +/- 0.56 vs LAD-control 0.07 +/- 0.60, p = 0.09 and RCA-LD 0.16 +/- 0.12 vs RCA-control 0.06 +/- 0.14, p = 0.002, respectively. The number of patients with additional ST elevation soon after reperfusion was LAD-LD 3 vs LAD-control 10, p < 0.05 and RCA-LD 0 vs RCA-control 8, p < 0.001, respectively.

Generalized dermal dysplasia and perifollicular mucinosis in a dog.

Multiple nodules 0.5-2 cm in diameter were observed in the dorsal skin of a 1.5-year-old male castrated mongrel dog. Histopathologically, perifollicular mucinosis and clusters of small cells with a dense, round nucleus and scant cytoplasm were seen. The small cell clusters were present in the dermis near the epidermal basal layer or surrounding hair follicles. These cells demonstrated different staining patterns for vimentin and keratin depending on their location; ultrastructurally, they were poor in organelles and connected to each other with cytoplasmic projections. These findings suggested that they were immature epithelial cells. The case was diagnosed as generalized dermal dysplasia and perifollicular mucinosis.

Factor VII Morioka (FVII L-26P): a homozygous missense mutation in the signal sequence identified in a patient with factor VII deficiency.

We investigated the molecular basis of factor VII deficiency in a Japanese patient and identified a novel missense mutation in the signal sequence of the gene. Factor VII activity and antigen level measured in the patient were 10.7% and 11% of normal, respectively. All exons except 1B and the 5'-flanking region containing promoter region were amplified by polymerase chain reaction (PCR) from genomic DNA. Sequencing analysis of the PCR fragments revealed that the patient was a homozygote for a T to C substitution at nucleotide position 38. This mutation predicts an amino acid replacement of leucine to proline at codon -26 in the hydrophobic core of the signal peptide, which probably affects translocation of the protein into endoplasmic reticulum and subsequently causes reduction in plasma factor VII level.

Purification and cDNA cloning of bovine liver 5'-nucleotidase, a GPI-anchored protein, and its expression in COS cells.

A glycosylphosphatidylinositol (GPI)-anchored protein, 5'-nucleotidase [EC 3.1.3.5], was released from the membrane of bovine liver by use of phosphatidylinositol-specific phospholipase C (PI-PLC) of Bacillus thuringiensis and purified by several column chromatographies to a homogeneous state. The purified protein has an apparent molecular mass of 61 kDa, as estimated by SDS-polyacrylamide gel electrophoresis. From the partial amino acid sequence of a tryptic peptide, mixed oligonucleotides were synthesized and used to screen a lambda gt11 liver cDNA library, and one positive clone, pE1, was isolated. Since the insert of the clone lacked the NH2-terminal coding region, another lambda gt11 liver cDNA library was screened by using a synthetic probe corresponding to the 5' region of the insert of pE1. Three additional cDNA clones were obtained. Sequencing of these cDNAs revealed an open reading frame that encodes a 574-residue polypeptide with a calculated mass of 63,084 Da. The predicted structure showed two highly hydrophobic stretches at both ends of the protein, like those of rat and human 5'-nucleotidases. The NH2-terminal 26 residues comprise a signal peptide and the COOH-terminal hydrophobic stretch may serve as a signal for the posttranslational GPI modification. An expression vector of the cDNA, pSVNT, was constructed in a mammalian expression vector pSVL and the 5'-nucleotidase activity was transiently expressed in COS-1 cells. The expressed activity was about 8 times higher than the pSVL-transfected control activity. PI-PLC released 45% of the transiently expressed 5'-nucleotidase activity, indicating that the cDNA isolated here encodes this enzyme expressed as a GPI-anchored protein.

[An adult case of anomalous origin of the left coronary artery from the pulmonary artery].

An adult case of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) was presented. A 37-year-old male was admitted for ventricular arrhythmia and chest pain. The catheterization, left ventriculography and coronary arteriography revealed ALCAPA with anterolateral ventricular aneurysm and mild mitral regurgitation. An operation using Takeuchi procedure was performed. The postoperative course was uneventful and the results of the catheterization, left ventriculography and coronary arteriography performed immediately and five years after the operation were excellent except mild supravalvular pulmonary stenosis. Six years later, the patient is asymptomatic and well.

[A case report of aortic stenosis complicated with complete atrioventricular block].

A case of aortic stenosis complicated with complete atrioventricular block preoperatively was reported. The patient was a 73-year-old woman who had been suffering from dyspnea and fatigability. Preoperative echocardiography revealed severe left ventricular hypertrophy, marked left ventricular-aortic pressure gradient (144 mmHg), and the impairment of left ventricular distensibility. Therefore, DDD pacemaker was implanted in addition to aortic valve replacement. Postoperative cardiac catheterization revealed that cardiac output was increased about 25% by DDD pacing compared with VVI pacing. It is more reasonable to employ DDD pacemaker to the case of which distensibility is impaired.

Gardenia fruit extract does not stimulate the proliferation of cultured vascular smooth muscle cells, A10.

We investigated the effect of a hot water extract from Gardenia fruit (Gardenia jasminoides Ellis) (GFE), which has a stimulatory effect on endothelial cell proliferation, on the proliferation of A10 cells, an established cell line of vascular smooth muscle cell from murine aorta in a culture system. GFE did not change the number of A10 cells after a 48 h culture. GFE significantly increased the incorporation of [3H]thymidine and [14C]leucine into the acid-soluble fraction of bovine aortic endothelial cell layers, but significantly decreased that of A10 cells. These results suggested that GFE stimulates the proliferation of endothelial cells but not of A10 cells. In the endothelial cell culture, GFE significantly increased the accumulation of basic fibroblast growth factor, which is an autocrine for endothelial cell proliferation in medium and low-affinity (glycosaminoglycans-binding) fractions, while A10 cells did not produce a significant amount of the factor. Since it is postulated that a selective stimulation of endothelial cell proliferation by increasing the production of basic fibroblast growth factor is appropriate for prevention of arteriosclerosis and thrombosis, GFE may contain a beneficial component as a useful drug.

Possible mechanism of the stimulatory effect of Artemisia leaf extract on the proliferation of cultured endothelial cells: involvement of basic fibroblast growth factor.

To investigate the possible mechanism of the stimulatory effect of a hot water extract from Artemisia leaf (Artemisia princeps PANPANINI) (AFE) on the proliferation of endothelial cells, cells from bovine aorta were cultured for 72 h in RPMI1640 medium supplemented with 10% fetal calf serum in the presence of 5 micrograms/ml AFE. The AFE treatment significantly increased the cell number after culture, while in the presence of 10 micrograms/ml unfractionated heparin, AFE conversely decreased it. This implied that AFE enhanced the cell growth promotion by basic fibroblast growth factor (bFGF). The accumulation of bFGF was significantly increased in the culture medium, in the low-affinity (glycosaminoglycans-binding) fraction, and in the cell extract fraction, but was unchanged in the high-affinity (receptor-binding) fraction. The contents of [35S]sulfate-labeled glycosaminoglycans in both cell layer and the medium were not increased by AFE treatment. The proliferation of A10 cells, an established cell line of smooth muscle cells from murine aorta, was not stimulated by AFE. A10 cells did not produce a significant amount of bFGF in the presence or absence of AFE. Thus, the production of bFGF was considered to be involved in AFE stimulation of cell proliferation. In conclusion, it was suggested that AFE stimulated endothelial cell proliferation by increasing the production of bFGF rather than by an increase in the number of bFGF receptors and the content of glycosaminoglycans in the cell layer. The enhanced reserve of bFGF in the low-affinity fraction of cell layer and in the medium would cause the AFE-stimulated proliferation of endothelial cells.

Gardenia fruit extract stimulates the proliferation of bovine aortic endothelial cells in culture.

We previously reported that Gardenia fruit extract had a stimulatory effect on fibrolysis in vitro. In the present study, we describe that the hot water extract from Gardenia fruit (GFE) stimulates the proliferation of cultured endothelial cells from bovine aorta. The cells were cultured in RPMI1640 medium supplemented 10% fetal bovine serum for up to 96 h in the presence of GFE (100 micrograms/ml and below). GFE at 25 and 50 micrograms/ml significantly increased the cell number after 48 h and above of the treatment. The incorporations of [3H]thymidine and [14C]leucine were significantly increased by 50 micrograms/ml GFE. The GFE stimulation of the proliferation was completely inhibited by 1 microM cycloheximide. The stimulatory activity of GFE was recognized in the low relative molecular mass (less than or equal to 10,000) fraction. In conclusion, it was demonstrated that a low molecular mass component of GFE stimulated endothelial cell proliferation and an increased protein synthesis was a essential component of this response. GFE may contain a useful compound to stimulate the proliferation of endothelial cells.

Heparin cofactor II inhibits thrombin-stimulated release of tissue plasminogen activator from cultured human endothelial cells in the presence of dermatan sulfate.

To investigate the involvement of heparin cofactor II (HC II) in fibrinolytic system, endothelial cells from human umbilical vein were cultured in the presence of HC II or antithrombin III (AT III) combined with or without thrombin. Although AT III significantly inhibited thrombin-induced increase in tissue plasminogen activator antigen (t-PA:Ag) release, HC II did not exhibit such a suppressive effect. In contrast, in the presence of dermatan sulfate, HC II inhibited thrombin stimulation of t-PA:Ag release more strongly than AT III did. The release of plasminogen activator inhibitor-1 antigen (PAI-1:Ag) was also stimulated by thrombin; this stimulation was inhibited only by the combination of HC II and dermatan sulfate. Comparatively high concentrations of HC II significantly suppressed thrombin stimulation of t-PA:Ag and PAI-1:Ag release but did not cause an obvious change of both release in the absence of thrombin. Based on these results, it was suggested that HC II may inhibit an increase in fibrinolytic activity mediated by thrombin-stimulated endothelial cells in the liquid phase through a suppression of thrombin stimulation of t-PA:Ag release, when plasma is exposed to vascular smooth muscle cells or fibroblasts which synthesize a significant amount of dermatan sulfate.

A stimulatory effect of Artemisia leaf extract on the proliferation of cultured endothelial cells.

To investigate the effect of the hot water extract from Artemisia leaf (Artemisia princeps Panpanini) (AFE) on the proliferation of endothelial cells, the cells from bovine aorta were cultured for up to 96 h in the presence of 1, 5, 10 or 50 micrograms/ml AFE in RPMI1640 medium supplemented with 10% fetal bovine serum. After a 72 h culture, the cell number was significantly increased by AFE at 1, 5 and 10 micrograms/ml. An increase in the cell number by 5 micrograms/ml AFE observed after a 72 or 96 h treatment. The incorporations of both [3H]thymidine and [14C]leucine by the growing cells were significantly increased by 5 micrograms/ml AFE after a 72 h treatment. In addition, the incorporation of [3H]thymidine by either growing or confluent cells was significantly increased by 50 micrograms/ml AFE after a 72 h treatment. The stimulatory activity of AFE was recognized in the low-molecular-weight fraction (molecular weight less than or equal to 10000 dalton). These results clearly indicated that AFE contained some low-molecular-weight component(s) which stimulates the proliferation of vascular endothelial cells in vitro.

[Intraoperative ST-T changes resembling myocardial ischemia due to accidental myocardial cooling].

During the mitral valve replacement on a 48-year-old male with mitral valve stenosis, ST-T changes (ST elevation and T inversion) suddenly appeared just before the start of cardiopulmonary bypass. Myocardial ischemia was suspected before we noticed the cardioplegic solution (about 10 degrees C) had been dripping on the surface of the right ventricle by mistake for about five minutes. After pouring warm saline over the myocardial surface, the EKG returned to normal within ten minutes. In this case, there appeared the elevation of the J point which is characteristic of profound hypothermia and is easily mistaken as myocardial ischemia because of ST segment elevation. It is very important to observe even a trivial intraoperative action which is not directly related with the operative procedure.

Intracoronary urokinase in acute myocardial infarction: prevalence of total coronary occlusion during the early hours, effects on myocardial infarct size and left ventricular function, and outcome of residual coronary stenosis.

The effects of intracoronary thrombolysis (ICT) were studied in 88 acute myocardial infarction patients. Total coronary occlusion was observed in 67 of the 88 patients (76.1%) who were evaluated within 6 hours of the onset of symptoms. Among these 67 patients 42 (62.7%) were successfully recanalized by intracoronary urokinase. The recanalization rate was higher in the lesion at the left anterior descending artery, in younger patients (49 years or less) and in patients with a shorter history of pre-infarction angina. Eight of 11 patients (72.7%) with subtotal coronary occlusion and 17 of 35 patients (48.6%) with recanalization after ICT showed spontaneous regression of the residual coronary stenosis at the chronic stage angiography. There was no re-occlusion in the subtotal occlusion group and only 6 cases of re-occlusion (17.1%) in the recanalization group. The majority of re-occlusions progressed from the lesion with 99% residual stenosis and delayed filling. Accordingly the true value of additional percutaneous transluminal coronary angioplasty would be limited to the latter cases. Reduction in infarct size and improvement in left ventricular function were limited to those patients with incomplete or subtotal coronary occlusion and were not seen in cases with total obstruction which was recanalized by ICT.

Serum levels of lipids and apolipoproteins in angiographycally defined ischemic heart disease.

To determine the predictive risk factors of the severity of the coronary artery disease, the serum levels of lipoprotein cholesterol and apolipoprotein were measured in 103 patients undergoing coronary angiography examination for suspected myocardial ischemia. The extent and severity of the coronary artery disease (CAD) were assessed by assigning scores to each lesion. Twenty-six female patients (59 +/- 8 yrs.) showed a stronger relationship of apo B and apo A-I/B to the coronary scores than the 77 male patients (57 +/- 8 yrs.). The male patients were divided into four groups based on the coronary scores: H-CAD (range: over 11 points), M-CAD (5-10 points), L-CAD (1-4 points) and N-CAD (0 point). The atherogenic risk factors other than the abnormalities in lipid metabolism (obesity index, fasting plasma glucose, smoking and blood pressure) were well matched in the four groups. T.C., LDL-C., HDL-C., HDL2-C., apo B, apo A-I/B ratio and apo A-II/B ratio significantly differed in the H-CAD and N-CAD groups. These results indicate that T.C., LDL-C., HDL-C., HDL2-C., apo B, apo A-I/B ratio and apo A-II/B ratio are predictive risk factors of the coronary heart disease. Furthermore, apo B and apo A-I/B ratio significantly differed in the H-CAD and L-CAD groups. These results suggest that apo B and apo A-I/B ratio may be good discriminators of the severity of the coronary heart disease.