Critical role of bone marrow apoptosis-associated speck-like protein, an inflammasome adaptor molecule, in neointimal formation after vascular injury in mice.

BACKGROUND: Inflammatory cytokines such as interleukin (IL)-1 beta and IL-18 play an important role in the development of atherosclerosis and restenosis. Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is an adaptor protein that regulates caspase-1-dependent IL-1 beta and IL-18 generation; however, the role of ASC in vascular injury remains undefined. Here, we investigated the contribution of ASC to neointimal formation after vascular injury in ASC-deficient (ASC(-/-)) mice. METHODS AND RESULTS: Wire-mediated vascular injury was produced in the femoral artery of ASC(-/-) and wild-type mice. Immunohistochemical analysis revealed that ASC was markedly expressed at the site of vascular injury. Neointimal formation was significantly attenuated in ASC(-/-) mice after injury. IL-1 beta and IL-18 were expressed in the neointimal lesion in wild-type mice but showed decreased expression in the lesion of ASC(-/-) mice. To investigate the contribution of bone marrow-derived cells, we developed bone marrow-transplanted mice and found that neointimal formation was significantly decreased in wild-type mice in which bone marrow was replaced with ASC(-/-) bone marrow cells. Furthermore, in vitro experiments showed that the proliferation activity of ASC(-/-) vascular smooth muscle cells was not impaired. CONCLUSIONS: These findings suggest that bone marrow-derived ASC is critical for neointimal formation after vascular injury and identify ASC as a novel therapeutic target for atherosclerosis and restenosis.

A case of mitochondrial cardiomyopathy with pericardial effusion evaluated by (99m)Tc-MIBI myocardial scintigraphy.

Heart failure is important in determining the prognosis of cardiomyopathy caused by mitochondrial gene abnormalities. We report herein the case of a patient with pericardial effusion and heart failure in whom mitochondrial cardiomyopathy was definitively diagnosed. A 56-year-old woman consulted her primary physician with exertional dyspnea. Examination revealed edema and pericardial effusion, and diuretics were prescribed. However, after marked left ventricular hypertrophy (LVH) was noted, she was admitted to our hospital for further evaluation. Further examination revealed short stature, ptosis, generalized muscle atrophy, and sensorineural hearing loss. Echocardiography showed LVH, a global decrease in wall motion, and pericardial effusion. Physical and laboratory findings, including glucose intolerance and elevated serum lactate, suggested mitochondrial cardiomyopathy. Genetic testing confirmed cardiomyopathy due to a mitochondrial a3243g mutation. After treatment to improve heart failure, marked washout was shown on (99m)Tc-MIBI (methoxyisobutylisonitrile) myocardial scintigraphy, suggesting a correlation with mitochondrial dysfunction.

Contemporary trend of reduced-dose non-vitamin K anticoagulants in Japanese patients with atrial fibrillation: A cross-sectional analysis of a multicenter outpatient registry.

BACKGROUND: Non-vitamin K antagonist oral anticoagulants (NOACs) have been widely used to prevent stroke in non-valvular atrial fibrillation (NVAF) patients. Stringent monitoring is not required for NOACs, albeit dose adjustments are needed based on specific patient factors, such as renal function, body weight and age, or concomitant medications. We investigated the NOAC dosing patterns and evaluated the predictors of the non-standardized dose reduction (NSDR). METHODS: A total of 2452 newly diagnosed NVAF patients were consecutively recruited from secondary- and tertiary-care hospitals between 2012 and 2017. The NOAC doses were classified as one of three: (1) full dose; (2) standardized dose reduction (SDR); or (3) NSDR, consistent with Japanese package inserts. RESULTS: Overall, 66.8% (N=1637) of the NVAF patients (median age: 69 years, interquartile range [IQR]: 60-76; 70% male; median CHA2DS2-VASc score of 2, IQR: 1-3) received NOACs. NOAC use dramatically increased during the study period (51.2% in 2012-13 to 74.4% in 2016-17). The percentages of SDR and NSDR were 19.6% and 14.4%, respectively; a proportion of SDR and NSDR did not alter drastically. Older age, concomitant antiplatelet therapy, impaired renal function, and prior heart failure or left ventricular dysfunction were independently associated with NSDR. Of note, patients with a high risk (CHA2DS2-VASc score ≥2) had the highest proportion of NSDRs. CONCLUSIONS: Nearly half of the NOAC dose reductions in our registry were deemed "non-standardized," which were seen mostly in patients at significant risk for ischemic stroke. The physician's apprehension regarding excessive bleeding under NOAC use should be appropriately balanced with concern for an increased risk of embolic events.

M-CSF accelerates neointimal formation in the early phase after vascular injury in mice: the critical role of the SDF-1-CXCR4 system.

OBJECTIVE: Since the macrophage colony-stimulating factor (M-CSF) has been shown to stimulate differentiation and proliferation of monocyte/macrophage lineage and to be involved in the process of neointimal formation after vascular injury, we tested the effects of M-CSF on the recruitment of bone marrow-derived progenitor cells in neointimal formation after vascular injury in mice. METHODS AND RESULTS: Wire-mediated vascular injury was produced in the femoral artery of C57BL/6 mice. Recombinant human M-CSF [500 microg/(kg x day)] or saline (control) was administered for 10 consecutive days, starting 4 days before the injury. Treatment with M-CSF accelerated neointimal formation in the early phase after injury, and this neointimal lesion mainly consisted of bone marrow-derived cells. M-CSF treatment had no effect on the mobilization of endothelial progenitor cells (EPCs: CD34+/Flk-1+) and reendothelialization after injury. The stromal cell-derived factor-1 (SDF-1) was markedly expressed in the neointima and media after injury, whereas CXCR4+ cells were observed in the neointima. Further, a novel CXCR4 antagonist, AMD3100, significantly attenuated the M-CSF-induced neointimal formation. CONCLUSIONS: These findings suggest that M-CSF accelerated neointimal formation after vascular injury via the SDF-1-CXCR4 system, and the inhibition of this system has therapeutic potential for the treatment of cardiovascular diseases.

Comparison of dietary triacylglycerol oil and diacylglycerol oil in protein kinase C activation.

The objective of the present study was to compare the effects of dietary diacylglycerol (DAG) oil with triacylglycerol (TAG) oil with a similar fatty acid composition (fatty acid chain range: C14-C22, C18 fatty acid chain: >90%) on protein kinase C (PKC) activation and on 1,2-DAG levels. Using male Wistar rats, no differences in cytosolic and membrane PKC activities in the lingual, esophageal, gastric, small intestinal, cecal, proximal colonic, and distal colonic mucosa were found between the 5% DAG and TAG oil groups, or between the 23% DAG and TAG oil groups after 1 month of feeding. The 1,2-DAG levels in the cecum and colon contents and in the feces and serum in male Wistar rats after a diet containing either 10% DAG or TAG oil feeding were similar between the groups. Moreover, exposure of Caco-2 cells to DAG and TAG oils had no effect on PKC activity in the membrane fraction, but 1,2-dioctanoyl glycerol composed of short-chain fatty acids (C8) did, suggesting the absence of an influence on PKC activity in DAG and TAG oils composed of long-chain fatty acids. In summary, the effects of DAG oil ingestion on PKC activity in the digestive tract and lingual mucosa, and on 1,2-DAG levels in the cecum and colon contents and in the feces and serum were similar to those observed for TAG oil ingestion.

Incomplete ventricular septal rupture following blunt chest trauma--a case report.

This report describes a case of traumatic incomplete rupture of the ventricular septum, a rare complication caused by blunt chest trauma. Although a serial ECG progressed its course similar to acute anteroseptal myocardial infarction in this case, there was little clinical clue of septal tear. The diagnosis was established by transthoracic echocardiography. The authors chose a conservative line of management rather than surgical repair for incomplete septal rupture because of the patent's stable clinical course and hemodynamic status. A sequence of echocardiography during a 32-day stay in the hospital showed no change in the extent of incomplete septal rupture, septal structure, systolic function, and shape of left ventricle and also obtained no evidence of shunting through the rupture. In conclusion, echocardiography is a useful investigation to make a diagnosis as well as for follow-up in case of incomplete ventricular septal rupture. A close follow-up of incomplete septal rupture with serial echocardiography should be performed, because several cases of delayed ventricular septal rupture following blunt chest trauma have been reported.

Metabolites of dietary triacylglycerol and diacylglycerol during the digestion process in rats.

The present study investigated the metabolic fate of dietary TAG and DAG and also their digestion products in the stomach and small intestine. A diet containing 10% TAG or DAG oil, enriched in 1,3-DAG, was fed to Wistar rats ad libitum for 9 d. After 18 h of fasting, each diet was re-fed ad libitum for 1 h. The weights of the contents of the stomach and small intestine were measured, and the acylglycerol and FFA levels were analyzed by GC at 0, 1, and 4 h after the 1-h re-feeding. The amounts of re-fed diet ingested and the gastric and small intestinal content were not different between the two diet groups. In the TAG diet group, the main products were TAG and DAG, especially 1(3),2-DAG. In addition, 1,3-DAG and 1(3)-MAG were present in the stomach, and the 1,3-DAG levels increased over time after the re-feeding period. In the DAG diet group, the main products in the stomach were DAG, MAG, FFA, and TAG. There were significantly greater amounts of 1,3-DAG, 1(3)-MAG, and FFA in the DAG diet group in the stomach compared with the TAG diet group. The amount of FFA in the stomach relative to the amount of ingested TAG plus DAG in the DAG diet group was higher than that in the TAG diet group. Acylglycerol and FFA levels were considerably lower in the small intestine than in the stomach. These results indicate that, in the stomach, where acyl migration might occur, the digestion products were already different between TAG and DAG oil ingestion, and that DAG might be more readily digested by lingual lipase compared with TAG. Furthermore, almost all of the dietary lipid was absorbed, irrespective of the structure of the acylglycerol present in the small intestine.

Nonanastomotic rupture of thoracic aortic Dacron graft treated by endovascular stent graft placement.

A 61-year-old man had a Stanford type A acute aortic dissection, and the total aortic arch was replaced with 22-mm knitted Dacron graft in 1996. In 2006, he underwent mitral valve replacement and tricuspid valve repair due to severe mitral and tricuspid valve regurgitation. Although preoperative computed tomography (CT) scan suggested pseudoaneurysm around the Dacron graft replaced with aortic arch, it could not be repaired concomitantly. Four months later, in view of the technical difficulties of an open surgical procedure, the prosthetic graft failure was repaired by endovascular stent graft consisting of a Gianturco Z stent covered with an UBE woven Dacron graft. However, during a follow-up, aneurysm sac diameter increased without any sings of endoleak in follow-up CT scans. Redo endovascular stent graft placement using a Gore-TAG device was performed. Subsequently, shrinkage of the pseudoaneurysmal sac could be observed.

Gigantic bronchial artery aneurysm treated with transcatheter arterial embolization: a case report.

Bronchial artery aneurysm (BAA) is a rare condition. Rupture of BAA can cause critical hemorrhage, and intervention for BAA is thus recommended. A 69-year-old woman presented with BAA 70 mm in diameter in the right hilum of the lung. Transcatheter arterial embolization for afferent arteries of the BAA was performed and the BAA has subsequently been shrinking as observed by radiography. We present this case and a brief review of management of BAA based on the literature.

Higher maximal serum concentration of methotrexate predicts the incidence of adverse reactions in Japanese rheumatoid arthritis patients.

Weekly pulsed low-dose methotrexate (MTX) is a standard regimen for rheumatoid arthritis (RA). Severe adverse reactions to MTX, such as pneumonia and cytopenia, sometimes occur; however, it is difficult to predict the development of these adverse reactions. In this article, we examine the serum concentrations of orally administered MTX of 69 Japanese patients with RA in the clinical setting. The maximum serum concentration (C (max)) after the first dose of the weekly administration and the time at which C (max) was obtained (T (max)) were analyzed. C (max) correlated with the administered dose before measurement. The average T (max) was 2.0 +/- 0.8 h, and none of the patients showed a T (max) of more than 4 h. In addition, we demonstrated that the weekly MTX dosage and the mean dosage of steroids were significantly higher in patients with adverse reactions than in those without them, and the C (max) after the first dose of the weekly administration particularly correlated with the incidence of adverse reactions (P < 0.001). In fact, the cut-off point of C (max) (0.16 micromol/l) was a sensitive predictor of the adverse reactions (sensitivity 81% and specificity 67%). We concluded that C (max) after the first dose of weekly administration is a useful parameter for predicting the development of adverse reactions to MTX.

Oxidative stress is involved in the development of experimental abdominal aortic aneurysm: a study of the transcription profile with complementary DNA microarray.

BACKGROUND: The role of oxidative stress in the formation of aneurysms is not fully understood. We used the complementary DNA (cDNA) microarray technique to determine the transcription profile in the development of elastase-induced abdominal aortic aneurysm in rat models, with an emphasis on the oxidative stress-related genes. MATERIALS AND METHODS: In the experimental group, rat abdominal aortas were perfused with elastase to induce AAA. In the control group, a sham operation was performed with perfusion of the aortas with saline solution. Four or five animals were used for each time point for each of the elastase-treated or saline-treated groups. At day 2, day 7, and day 10 after surgery, the external aortic diameter was measured and AAA formation was estimated. Total RNA was isolated from aortas and subjected to cDNA microarray analysis with the use of the rat genome U34A high-density oligonucleotide DNA chip (Affymetrix, Santa Clara, Calif), which contains a total number of 8799 genes of which 2017 are expressed sequence tag (EST) genes. The data were analyzed with the GENECHIP Data Mining Tool software (Affymetrix). For genes of interest, reverse-transcription polymerase chain reaction was performed to confirm their expression level. RESULTS: Comparison ranking analysis revealed that during AAA development, the expression of 212 genes, including 46 of EST genes, increased by more than two-fold and 229 genes, including 95 of EST genes, decreased by more than two-fold in at least one of the three time points. The regulated genes included those encoding heme oxygenase, inducible nitric oxide synthase, some extracellular matrix proteins, members of the matrix metalloproteinase family, and those associated with prooxidant/antioxidant and inflammatory responses. Reverse-transcription polymerase chain reaction analysis confirmed the upregulation of genes involved in oxidative stress, such as heme oxygenase, inducible nitric oxide synthase, 12-lipoxygenase, and heart cytochrome c oxydase subunit VIa, and the downregulation of antioxidant genes, such as superoxide dismutase, reduced nicotinamide adenine dinucleotide-cytochrome b-5 reductase, and glutathion S-transferase. CONCLUSION: The cDNA microarray technique was useful for investigation of the transcription profiles during the development of AAA. Our results indicate that oxidative stress may play a pivotal role in the pathologic progression of AAA.

Induction of apoptosis in vascular smooth muscle cells by mechanical stretch.

We studied the response of porcine vascular smooth muscle cells (PVSMCs) to cyclic sinusoidal stretch at a frequency of 1 Hz. Cyclic stretch with an area change of 25% caused an increase in PVSMC apoptosis, which was accompanied by sustained activation of c-Jun NH(2)-terminal kinases (JNK) and the mitogen-activated protein kinase p38. Cyclic stretch with an area change of 7% had no such effect. Infection of PVSMCs with recombinant adenoviruses expressing constitutively active forms of upstream molecules that activate JNK and p38 also led to apoptosis. The simultaneous blockade of both JNK and p38 pathways with adenovirus-mediated expression of dominant-negative mutants of c-Jun and p38 caused a significant decrease (to 1/2) of the apoptosis induced by 25% cyclic stretch. The 25% stretch also caused sustained clustering of tumor necrosis factor-alpha (TNF-alpha) receptor-1 and its association with TNF-alpha receptor-associated factor-2 (TRAF-2). Overexpressing the wild-type TRAF-2 in PVSMCs caused an increase in apoptosis. In contrast, the expression of a dominant-negative mutant of TRAF-2 attenuated stretch-induced apoptois. These results support the hypothesis that circumferential overload under hypertensive conditions induces a clustering of death receptors that cause vascular smooth muscle cell apoptosis.