Expression of the cyclin-dependent kinase inhibitor, p21Waf-1/Cip-1/Sdi-1, in human vascular smooth muscle cells in the proliferating state

Excess proliferation of vascular smooth muscle cells (SMC) is an important aspect of atherogenesis. Cell-cycle regulatory proteins such as cyclin and cyclin-dependent kinases are vital for cell-cycle progression. To understand the role of the cyclin-dependent kinase inhibitor, p21Waf-1/Cip-1/Sdi-1 (p21Waf-1), on human atherogenesis, we tested p21Waf-1 expression in human atherosclerotic lesions and cultured SMC. Immunohistochemical staining revealed that SMC in neointimal lesions expressed p21Waf-1. No evidence of the p53 protein could be detected. By Western blotting, cultured SMC obtained from a neonate revealed that a higher level of p21Waf-1 expression correlated with a higher expression of proliferating cell nuclear antigen and a lower expression of the contractile protein than that observed in cells obtained from aged donors. When the phenotypes of SMC were changed by modification of serum concentration and cell densities, p21Waf- expression was maximal in serum-stimulated SMC at low cell densities despite the low expression of p53. Furthermore, serum stimulation transiently increased the p21Waf-1 expression of quiescent SMC, which was synchronized with the transition from the G0/G1 to the S phase as well as with cyclin D1 expression. These results may suggest that the negative regulator of cell-cycle progression also plays a role in regulating the appropriate cell-cycle progression in SMC. Growth stimuli may induce both growth-promoting and growth-inhibitory factors in SMC. The balance between these two opposing factors may play an important role in the determination of cell-cycle progression.     

Novel method of displaying coronary CT angiography: Angiographic view.

Background A method of displaying coronary computed tomography (CT) angiography, which enables evaluation of coronary artery disease (CAD) with fewer images and is understandable to the third person, is preferable. Methods and Results A maximum intensity projection image was created in which contrast media in the ventricles is eliminated, enabling an overview of CAD in a single 3-dimensional (D) image that can be rotated to be viewed at various angles and is easily understood by a third person. Conclusions A novel method of displaying coronary CT angiography in a single 3-D image has been developed and we believe it should become available for many workstations.     

Saprophytic mycosis appearing as a necrotic bronchial tumor and masking lung cancer]

We describe four cases of saprophytic mycosis superficially covering lung cancer to form a bronchial necrotic tumor. Although the first biopsy in each case disclosed mycosis with necrosis, repeated transbronchial biopsy revealed malignant cells. In two of the four cases, we started treatments for lung cancer based on the clinical diagnosis preceding histological diagnosis. Although no treatment to eliminate fungi was performed, all cases showed an uneventful clinical course. It is important not to misdiagnose cancer as a fungal disease on the basis only of a transbronchial biopsy, and to bear in mind this type of saprophytic mycosis.     

Observation of the ischemic cascade in humans using contrast echocardiography during dobutamine stress.

Experimental studies have postulated the ischemic cascade and the present study was designed to elucidate whether it can be observed in the clinical setting. Fifty-three patients suspected of having coronary artery disease were studied. Myocardial perfusion abnormalities (MPA) and wall motion abnormalities (WMA) were assessed simultaneously by infusion of Levovist during dobutamine stress echocardiography. Time - intensity data of myocardial opacification were fitted for Y=A (1-e(-)(beta) (t)) from which the rate of increase (beta) of intensity were derived both at rest and during stress. Wall motion was also given a score. Bright opacification was observed in 50 patients: 25 showed significant stenosis (>50%) in the left anterior descending artery (group II) on coronary angiography and 25 did not (group I). Significant differences were found in the beta ratio (stress/rest) between the 2 groups at a low-dose (2.0+/-0.3 vs 1.5+/-0.5, p<0.05) and at a high-dose of dobutamine (2.7+/-1.0 vs 1.1+/-0.5, p<0.001), whereas the wall motion score differed only at a high-dose. Of the 25 patients in group II, MPA preceded WMA in 12, both occurred at the same stage in 12, and neither MPA nor WMA was seen in 1. These data prove the ischemic cascade clinically, using contrast echocardiography, by demonstrating that MPA precede WMA during dobutamine stress in patients with coronary stenosis.     

Double SCN5A mutation underlying asymptomatic Brugada syndrome

OBJECTIVES: The purpose of this study was to identify risk markers in patients with Brugada syndrome. BACKGROUND: Patients with Brugada syndrome who experience syncope or aborted sudden death are at high risk for recurrent lethal arrhythmias. The prognosis and therapeutic approaches in asymptomatic individuals with a Brugada-type ECG (asymptomatic Brugada syndrome) are controversial. METHODS: We genetically screened 30 asymptomatic probands (29 men and 1 woman; mean age 47.1 years) exhibiting a spontaneous Brugada-type ECG. Family members of patients with Brugada syndrome were excluded from the study. RESULTS: Twenty-nine of 30 patients (96.7%) remained symptom-free for at least 3 years. One patient (case 1) with a family history of sudden death died suddenly during sleep. Ventricular fibrillation was induced by programmed electrical stimulation in 14 of 18 subjects (78%), but none of these 18 subjects developed spontaneous ventricular arrhythmias. Genetic screening failed to identify SCN5A mutations in most cases but demonstrated a novel double missense mutation (K1527R and A1569P) located on the same allele in another asymptomatic subject (case 2). Heterologously expressed mutant Na channels exhibited a negative shift of steady-state inactivation (9.2 mV) and enhanced slow inactivation, suggesting this individual harbors a subclinical channel dysfunction compatible with symptomatic Brugada syndrome. CONCLUSIONS: Asymptomatic individuals with a Brugada-type ECG generally have a better prognosis than their symptomatic counterparts, but a subgroup of these individuals may have a poor prognosis. Severe Na channel dysfunction as a result of SCN5A mutations may not be sufficient to cause symptoms or arrhythmias in patients with Brugada syndrome, suggesting unknown factors or modifier genes influence arrhythmogenesis.     

Attenuation of graft injury in porcine liver transplantation from non-heart beating donor by FR167653

BACKGROUND/AIMS: Orthotopic liver transplantation (OLTx) from non-heart beating donor (NHBD) often involves hepatic warm ischemia and reperfusion injury which is triggered by the inflammatory cytokines. This study was carried out to investigate whether a newly synthesized cytokine suppressive anti-inflammatory agent, FR167653, attenuates graft injury in OLTx from NHBD. METHODOLOGY: Porcine OLTx from NHBD was performed. No-heart beating time was scheduled to be 60 minutes. Animals were divided into two groups: no treatment control (CT) group (n=5), and FR167653 treated (FR) group (n=5), in which FR167653 was administered intravenously before the aortic cross clamp in the donor, and before and after the hepatic allograft reperfusion in the recipient continuously. RESULTS: Four out of five pigs died within 24 hours and one on postoperative day 1 from graft liver failure in the CT group, while two pigs died on day 3, and three survived more than 7 days in the FR group (p<0.05). Microcirculatory disturbance was attenuated, liver injury was lessened, and ATP resynthesis was enhanced in the FR group. Additionally, FR167653 inhibited neutrophils infiltration in the liver tissue, and suppressed release of inflammatory cytokines after OLTx from NHBD. CONCLUSIONS: The treatments with FR167653 successfully prevented graft injury after OLTx from NHBD by means of improvement of liver microcirculation, and attenuation of neutrophils activation. The inhibitory effect of FR167653 on the release of inflammatory cytokines played an important role in the liver graft protection.     

Non-specific reactions in four methods measuring (1-->3)-beta-D-glucan levels in plasma

We detected non-specific reactions in the measurement of (1-->3)-beta-D-glucan levels (beta-glucan) in plasma, and the influences of the non-specific reactions on sensitivity and specificity of measurement methods were examined. In this study, 460 plasma samples from 174 patients at Kawasaki Medical School Hospital were used, and the plasma beta-glucan levels were measured by different four methods. The methods included the dilution-heating-endpoint (DHE), dilution-heating-turbidimetric (DHT), alkaline-kinetic (AK), and alkaline-endpoint method (AE) with and without 4-amidinophenyl benzoate hydrochloride (APB) of a protease inhibitor blocking the Limulus reaction. Non-specific reactions were detected from the calculated value under conditions with APB. Therefore, both of the actual values and the values equivalent to non-specific reactions were calculated. The incidence of non-specific reactions was 2.4% in DHE method, 0% in DHT, 53.3% in AK, and 99.3% in AE. The sensitivity and specificity in the methods were 35.7% and 96.0%, 28.6% and 96.0%, 78.6% and 80.1%, and 57.1% and 84.1%, respectively. When subtracted the non-specific reaction values from the actual values in AK and AE method, the specificity was increased by 91.4% and 94.0%, respectively. In these two methods, the non-specific reaction was considered to be a major cause of the low specificity. Finally, to measure plasma beta-glucan levels accurately, non-specific reactions should be excluded as possible by further improvement of measurement methods.     

Successful Low-Dose Chemotherapy Using Vinblastine and Methotrexate for the Treatment of an Ileoanal Pouch Mesenteric Desmoid Tumor: Report of a Case

The purpose of this report was to describe the first known case of ileoanal pouch salvage by a low-dose regimen of vinblastine and methotrexate chemotherapy for the treatment of desmoid tumor arising from the mesentery of the ileoanal pouch in a patient who had undergone ileal pouch-anal anastomosis for familial adenomatous polyposis. Mesenteric desmoid tumor involving the ileoanal pouch in a 28-year-old female was treated with vinblastine and methotrexate biweekly for 12 months and monthly for 12 months in an outpatient unit. The desmoid tumor response to the treatment was assessed at routine intervals by physical examination and magnetic resonance imaging. Desmoid tumor was successfully treated with a low-dose regimen of vinblastine and methotrexate chemotherapy without significant side effects, and function of the ileoanal pouch was fully preserved. Magnetic resonance imaging showed a decrease in desmoid tumor size and cellularity, and changes consistent with fibrosis. This is a unique case highlighting the possibility of ileoanal pouch salvage by low-dose combination chemotherapy using vinblastine and methotrexate in a familial adenomatous polyposis patient with mesenteric desmoid tumor.