Transglutaminase 2 has opposing roles in the regulation of cellular functions as well as cell growth and death.

Transglutaminase 2 (TG2) is primarily known as the most ubiquitously expressed member of the transglutaminase family with Ca(2+)-dependent protein crosslinking activity; however, this enzyme exhibits multiple additional functions through GTPase, cell adhesion, protein disulfide isomerase, kinase, and scaffold activities and is associated with cell growth, differentiation, and apoptosis. TG2 is found in the extracellular matrix, plasma membrane, cytosol, mitochondria, recycling endosomes, and nucleus, and its subcellular localization is an important determinant of its function. Depending upon the cell type and stimuli, TG2 changes its subcellular localization and biological activities, playing both anti- and pro-apoptotic roles. Increasing evidence indicates that the GTP-bound form of the enzyme (in its closed form) protects cells from apoptosis but that the transamidation activity of TG2 (in its open form) participates in both facilitating and inhibiting apoptosis. A difficulty in the study and understanding of this enigmatic protein is that opposing effects have been reported regarding its roles in the same physiological and/or pathological systems. These include neuroprotective or neurodegenerative effects, hepatic cell growth-promoting or hepatic cell death-inducing effects, exacerbating or having no effect on liver fibrosis, and anti- and pro-apoptotic effects on cancer cells. The reasons for these discrepancies have been ascribed to TG2's multifunctional activities, genetic variants, conformational changes induced by the immediate environment, and differences in the genetic background of the mice used in each of the experiments. In this article, we first report that TG2 has opposing roles like the protagonist in the novel Dr. Jekyll and Mr. Hyde, followed by a summary of the controversies reported, and finally discuss the possible reasons for these discrepancies.

Molecular mechanism by which acyclic retinoid induces nuclear localization of transglutaminase 2 in human hepatocellular carcinoma cells.

Nuclear accumulation of transglutaminase 2 (TG2) is an important step in TG2-dependent cell death. However, the underlying molecular mechanisms for nuclear translocation of TG2 are still poorly understood. In this study, we demonstrated that acyclic retinoid (ACR) induced nuclear accumulation of TG2 in JHH-7 cells, a hepatocellular carcinoma (HCC) leading to their apoptosis. We further demonstrated molecular mechanism in nuclear-cytoplasmic trafficking of TG2 and an effect of ACR on it. We identified a novel 14-amino acid nuclear localization signal (NLS) (466)AEKEETGMAMRIRV(479) in the 'C' domain and a leucine-rich nuclear export signal (NES) (657)LHMGLHKL(664) in the 'D' domain that allowed TG2 to shuttle between the nuclear and cytosolic milieu. Increased nuclear import of GAPDH myc-HIS fused with the identified NLS was observed, confirming its nuclear import ability. Leptomycin B, an inhibitor of exportin-1 as well as point mutation of all leucine residues to glutamine residues in the NES of TG2 demolished its nuclear export. TG2 formed a trimeric complex with importin-α and importin-ß independently from transamidase activity which strongly suggested the involvement of a NLS-based translocation of TG2 to the nucleus. ACR accelerated the formation of the trimeric complex and that may be at least in part responsible for enhanced nuclear localization of TG2 in HCC cells treated with ACR.

Evaluation of average amount of cerebral blood flow measured by brain perfusion index in patients with neuropsychiatric systemic lupus erythematosus.

We used the brain perfusion index (BPI), an indicator of the average amount of cerebral blood flow (CBF), to evaluate the usefulness of the average amount of CBF for neuropsychiatric systemic lupus erythematosus (NPSLE). Of the seventy three SLE patients examined in this study (total 100 scans), 16 patients (23 scans) had already been diagnosed with NPSLE based on clinical symptoms indicative of central nervous system involvement. In addition, 12 patients (17 scans) exhibited the antiphospholipid antibody syndrome (APS). BPI is significantly influenced by age and we therefore used the BPI ratio (ratio of age predicted BPI to measured BPI value) for each assessment. The mean BPI value of 100 scans was 11.2 +/- 2.79, and the mean BPI ratio was 0.99 +/- 0.24 in all SLE patients. The mean BPI ratio among NPSLE (0.84 +/- 0.19) was significantly lower than that of the non-NPSLE patients (1.04 +/- 0.24) (P < 0.0005). However, there was no difference in the mean BPI ratio between APS patients (0.98 +/- 0.24) and non-APS patients (0.99 +/- 0.25). These results indicate that the mean CBF assessed by the BPI ratio using SPECT is of use in the evaluation of central nervous system involvement in SLE patients.

Relationship between coronary blood flow velocity waveform and transmural distribution of myocardial blood flow in coronary artery.

It is important to know the transmural distribution of myocardial blood flow in assessing the severity of ischemia in coronary heart disease. We analyzed the relation between phasic waveform of epicardial coronary flow velocity with a Doppler flow probe in the left anterior descending artery in dogs and regional myocardial blood flow using a colored microsphere technique. Time-velocity integral in an average of 5 cardiac cycles was measured as an index of coronary blood flow during diastole (TVId) and systole (TVIs). The diastolic fraction of coronary blood flow (%DF) was defined as TVId/(TVId + TVIs). Myocardial specimens were divided into inner (subendocardial), middle, and outer (subepicardial) layers, and the inner layer to outer layer myocardial blood flow ratio (endo/epi ratio) was used as an index of transmural distribution of myocardial perfusion. The mean endo/epi ratio and the mean %DF decreased as the pressure gradient increased. There was a moderate but significant correlation (r = 0.57) between the endo/epi ratio and the %DF. In conclusion, analysis of the phasic pattern of coronary blood flow velocity provides some information about the transmural distribution of blood flow in the myocardium. The %DF may be a useful index for evaluating subendocardial ischemia.

Disturbance of consciousness associated with hypophosphatemia in a chronically alcoholic patient.

A 69-year-old man with chronic alcoholism was admitted to our hospital due to disturbance of consciousness and oliguria. Emergency laboratory examination revealed metabolic acidosis, hypoglycemia, hyponatremia, mild liver dysfunction, acute renal failure and rhabdomyolysis. After administration of fluids and nutrients and continuous hemodiafiltration, he recovered from all signs and symptoms except for disturbance of consciousness after 7 days. Since severe hypophosphatemia persisted, we administered adequate phosphates, and then his level of consciousness normalized. We discuss the relationships among alcohol abuse, hypophosphatemia and disturbance of consciousness, and recommend that hypophosphatemia be considered a potential cause of disturbance of consciousness in alcoholic patients.

[Measurement of coronary flow reserve using adenosine 5'-triphosphate in dogs].

Adenosine 5'-triphospate (ATP) was compared with adenosine and papaverine for the measurement of coronary flow reserve in 12 anesthetized dogs. Intracoronary bolus injection of ATP (1 ml, 1-500 microM) produced a dose dependent increase in the blood flow of the left anterior descending artery, which attained the plateau at the dose of 100 microM. The ratio of peak to resting coronary flow volume (coronary flow reserve) with 100 microM of ATP (3.5 +/- 0.5) was similar to that with 200 microM of adenosine (4.0 +/- 0.7) and 50 mM of papaverine (3.7 +/- 0.8). Hemodynamic variables did not change after administration of each drug, except left ventricular regional wall motion abnormality during papaverine injection. The coronary flow reserve as measured after intracoronary ATP administration (100 microM) decreased as the grade of stenosis of the left anterior descending artery progressed. In addition, the flow reserve was similar to that of adenosine or papaverine administration at each stenosis grade. Intravenous administration of ATP (1,000 micrograms/min) caused a similar increase in coronary blood flow as intracoronary ATP injection (100 microM). However, premedication with 8-phenyltheophylline, an adenosine receptor blocker, significantly suppressed the coronary dilatory effect of intravenous ATP and intracoronary adenosine but not the effect of intracoronary ATP. These results indicate that intracoronary ATP is useful for measuring coronary flow reserve and that its coronary dilatory effect is not mediated by metabolysis to adenosine.

[Evaluation of 123I-MIBG clearance from the myocardium; comparison of two methods--SPECT & planar methods].

In 123I-metaiodobenzylguanidine (MIBG) scintigram, MIBG clearance from the heart is used to evaluate the severity of various heart diseases. There are two methods for calculating MIBG clearance. One involves planar images (planar method) and the other uses a bull's eye map (SPECT method). In 158 patients and 10 normal subjects, we compared these two methods. Fifteen minutes and 4 hours after intravenous injection of 111 MBq MIBG, planar images and SPECT images were obtained. Then clearance from the heart was calculated by each method. Abnormal increase was defined as present if clearance was more than the mean + standard deviation of 10 normal subjects. Then, we examined the sensitivity with which each method could detect clearance abnormality in 158 patients. Thirty-two patients showed abnormality only on SPECT images, while planar images alone showed abnormalities in only 5 patients. The reason the SPECT method was more sensitive than the planar method may be as follows; in the case of decreased MIBG clearance from the lung, for example, in congestive heart failure, clearance by planar method is apparently decreased. Thus, the SPECT method can detect clearance abnormality more sensitively than the planar method, and if we evaluate MIBG clearance from the heart by the planar method, we must take into account MIBG clearance from the lung.

[Angle of defect on 123I-MIBG myocardial SPECT].

The quantitative assessment of infero-posterior defect on 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy was studied in 150 patients with non-ischemic heart diseases at rest. The bull's eye map, obtained from the SPECT images at 4 hours after MIBG injection, was evaluated by generating the blacked out map which exhibits regions with reduced % uptake under mean-2 SD of 13 normal controls. The blacked out regions involved infero-posterior segments and were closely resembled to the sector form. The central angle of this sector, which was named as angle of defect (AOD), significantly correlated with both the heart-to-mediastinum activity ratio (H/M) and the myocardial clearance of MIBG. Because H/M and clearance are widely used as quantitative indices in MIBG myocardial scintigraphy, these results indicate that AOD can also be used as a quantitative index of abnormal cardiac sympathetic nervous function, which is likely to appear in infero-posterior regions in non-ischemic cardiac diseases.

Estimation of coronary flow reserve with the instantaneous coronary flow velocity versus pressure relation: a new index of coronary flow reserve independent of perfusion pressure.

Our goal was to develop a new method to evaluate coronary flow reserve independent of perfusion pressure by equipping dogs with a Doppler flow probe in the left anterior descending artery. The slope of the linear portion of the velocity-pressure loop in the late diastolic phase was defined as VP slope; the hyperemic:basal ratio of the VP slope was defined as VP slope ratio, and the hyperemic:basal ratio of the mean coronary blood flow velocity was defined as coronary flow reserve (CFR). We measured VP slope ratio and CFR, altering aortic pressure by aortic banding and inferior vena cava occlusion. VP slope ratio was independent of aortic pressure, but CFR increased with increments in aortic pressure. The VP slope ratio and CFR decreased in the presence of coronary stenoses. In conclusion, VP slope ratio is considered to be a physiologic index of the severity of coronary stenosis, which is independent of perfusion pressure. This measure is easily applicable in clinical practice.

Investigation of the relationship between regression of hypertensive cardiac hypertrophy and improvement of cardiac sympathetic nervous dysfunction using iodine-123 metaiodobenzylguanidine myocardial imaging.

Although many theories exist on the subject, the mechanisms responsible for a reduction of hypertensive cardiac hypertrophy in response to antihypertensive therapy are still unclear. In order to investigate the relationship between regression of hypertensive cardiac hypertrophy and cardiac nervous function, we studied ten patients with untreated essential hypertension (six men and four women, 62+/-12 years old). Both echocardiography and iodine-123 metaiodobenzylguanidine (MIBG) myocardial imaging were performed before and after antihypertensive therapy. Left ventricular mass (LVM) was significantly reduced in conjunction with the reduction of blood pressure following treatment. MIBG myocardial images showed that the heart-to-mediastinum activity ratio (H/M) was significantly increased while the washout ratio was significantly decreased. Patients were divided into two groups according to the ratio of the LVM values before and after therapy (LVM ratio). Patients with an LVM ratio of less than 0.75 were classified as group A and those with values higher than 0.75 as group B. Neither the change in blood pressure nor the length of treatment was significantly different between these two groups. On the other hand, both the increase in H/M and the decrease in the washout ratio were significantly greater in group A than in group B. These results indicate that an improvement in cardiac sympathetic nervous function may be related to the regression of hypertensive cardiac hypertrophy. Increasing the subject base in these studies and a more precise analysis of the relevance of the data obtained from MIBG myocardial images are recommended to clarify how changes in cardiac sympathetic nervous function relate to the regression of hypertensive cardiac hypertrophy.

[A case of hypertensive hypertrophy in which both regression of hypertrophy and improvement of the abnormalities in iodine-123-metaiodobenzylguanidine (MIBG) myocardial imagings were observed after antihypertensive therapy].

A case of hypertensive hypertrophy is described in which both regression of hypertrophy and improvement of the abnormalities in iodine-123-metaiodobenzylguanidine (MIBG) myocardial imagings were seen after 7 months of antihypertensive therapy. A 58-year-old man was diagnosed as having essential hypertension and hypertensive hypertrophy. The patient was treated with antihypertensive drugs and showed regression of left ventricular hypertrophy on electrocardiograms and echocardiograms. MIBG observations made before and after antihypertensive therapy showed increased heart-to-mediastinum activity ratio and decreased cardiac washout ratio. Despite the many theories addressing the mechanisms of regression of left ventricular hypertrophy, the process is still unclear. In the present case, the improvement of cardiac sympathetic nervous dysfunction might have been related to the regression of left ventricular hypertrophy because the abnormality in MIBG images improved. MIBG, therefore, may be helpful in clarifying the mechanisms of the regression of hypertensive hypertrophy.

The efficacy of indomethacin in the treatment of uremic pericarditis.

Uremic pericarditis is common in patients undergoing chronic hemodialysis and has been difficult to cure using conservative medical and surgical methods of treatment. Indomethacin therapy for uremic pericarditis has been reported infrequently. We present a case of uremic pericarditis in which indomethacin was particularly effective. The patient was a 45-year-old woman with thrombocytopenia due to Banti's syndrome and congestive heart failure with pericardial effusion. She had been undergoing hemodialysis for 3 years for chronic renal failure due to chronic glomerulonephritis. Uremic pericarditis was diagnosed based on serological laboratory results, aspirated pericardial fluid, and an echocardiogram. She was treated with oral doses of 25 mg t.i.d. indomethacin in addition to hemodialysis and extracorporeal ultrafiltration method. Pericardial effusion and left ventricular dysfunction disappeared after indomethacin therapy for 38 days. There are conflicting reports on the efficacy of indomethacin therapy in patients with uremic pericarditis. While indomethacin therapy was very effective in our case, there may be many causes of uremic pericarditis for which indomethacin may not be efficacious. Further investigations of indomethacin therapy for patients with uremic pericarditis are necessary to elucidate the therapeutic mechanism by which indomethacin acts.

Hypersensitivity to heparin; a case report.

Hypersensitivity reactions to heparin are very rare. A generalized hypersensitivity reaction including as fever and skin rash to a porcine- and bovine-derived heparin preparation was observed in a hemodialysis patient due to the nephrotic syndrome. The patient revealed peripheral eosinophilia and normal serum IgE. The results of a drug lymphocyte stimulating test on heparin were positive. Following prednisolone administration and infusion of nafamostat mesilate as anticoagulant therapy during hemodialysis, the high fever and generalized urticaria disappeared. Caution is required when conducting heparin therapy on hemodialysis patients.

Case report: left ventricular apical hypertrophy in progressive limb-girdle muscular dystrophy.

Cardiac involvement is uncommon in patients with limb-girdle muscular dystrophy. This report describes a patient in whom concentric hypertrophy localized to the apical left ventricle was revealed during a long clinical course of skeletal muscular dystrophy, with evolving electrocardiographic changes also compatible with apical hypertrophic cardiomyopathy. Endomyocardial biopsy revealed similar histologic changes in the skeletal muscle biopsy specimen, characterized by muscle fiber atrophy and hypertrophy with a mild degree of interstitial fibrosis. The pathogenesis of cardiac hypertrophy in this case is unclear. However, the pathologic findings suggest that the myocardium may be involved in the same dystrophic process as the skeletal muscles.

[A successfully treated case of acute renal failure due to acute immune hemolytic anemia and nontraumatic rhabdomyolysis induced by streptomycin reinjection].

A case of acute immune hemolytic anemia associated with non-traumatic rhabdomyolysis (NTR) induced by streptomycin (SM) reinjection, which developed acute renal failure, has been reported. A 70-year-old female was admitted to our hospital because of sudden macroscopic hematuria after reinjection of 1g. SM. Laboratory findings on admission were as follows; hemoglobin and myoglobin were positive in urine. RBC 129 x 10(4)/microliters, Hb 4.9g/dl, Ht 11.1%, reticulocytes 52/1000, serum indirect billirubin 3.8g/dl, LDH 9, 230 WU, BUN 149mg/dl, Cr 7.9mg/dl, myoglobin 1, 400ng/ml and haptoglobin 10.6mg/dl. The drug lymphocyte stimulating test of SM was positive (215%). A direct antiglobulin test was also positive. An indirect antiglobulin test was negative, but became positive after incubation with SM. These observations made the diagnosis of SM-induced hemolytic anemia associated with NTR. On the second hospital day she developed anuria, and was put on hemodialysis treatment. Two months after the acute hemolytic episode and acute renal failure she recovered and is presently in good health without recurrence.