We previously identified a novel rat gene, Bdm2 (brain developmentrelated molecule 2), that was expressed in the brain predominantly during embryogenesis. To characterize BDM2 protein, we prepared glutathione Stransferase-BDM2 fusion protein (GST-BDM2) and raised antibody against this recombinant protein. This antibody recognized two bands of 36 kDa and 38 kDa on Western blotting of brain proteins. A 36-kDa protein band was detected on embryonic day 16 and the intensity of this band gradually decreased during subsequent embryonic development. The 38-kDa protein band was detected after birth and the intensity of this band increased as the brain matured.In vitrotranscription and translation of Bdm2 cDNA produced proteins of apparent molecular masses of 36, 38, and 49 kDa. Therefore, translation may be initiated at two different sites on Bdm2 mRNAin vivodepending on the developmental stage. When an expression construct of green fluorescent protein (EGFP) -Bdm2 was transfected into PC 12 cells, EGFP-BDM2-specific fluorescence was distributed in the cytoplasm, and was observed in the cell body and its processes after neuronal differentiation. Cell fractionation and immunoblot analysis of the embryonic brain also showed that BDM2 was distributed mainly in the cytoplasmic fraction. Recombinant BDM2 was phosphorylated by protein kinase Cin vitro. These results suggest that the two Bdm2 gene products may be differentially expressed in the developing and mature brain.

View full abstract

Impotence commonly occurs after events such as acute myocardial infarction, coronary bypass, head trauma, and cerebral bleeding, including subarachnoid hemorrhage. One possible explanation is that the hypoxia that occurs with such events results in damage to the testis or to the blood-testis barrier. We examined the effect of cardiac arrest in mice on testis weight and on the permeability of the blood-testis barrier to albumin and tumor necrosis factor alpha (TNF-α) . Testis weight was decreased by about 24% 12 h after cardiac arrest but had recovered fully by day 7. The testis/serum ratio for albumin also was increased 12 h after arrest showing a disruption in the blood-testis barrier with recovery by 24 h. There were no statistically significant effects on the permeability of the blood-testis barrier to TNF-α. These results show that cardiac arrest transiently affects testis weight and the integrity of its blood-barrier. Such transient changes might induce long-term effects on testicular function that could lead to impotence.

View full abstract

The aim of this study was to evaluate the role of the KATP channel in the mechanism of ischemic preconditioning in whole body rat hearts using a KATP channel opener (KRN2391) and KATP channel blockade (glibenclamide) . In protocol 1, seven groups of rats were studied : a control group, animals that received Ischemic preconditioning (3 min of left coronary artery occlusion and 5 min of reperfusion, repeated twice sequentially) prior to 30 or 60 min of ischemia and following 60 min reperfusion (IP30, IP60), animals that received pharmacological preconditioning (injection of KRN2391) prior to ischemia (PP30, PP60), and animals that were not preconditioned prior to ischemia (NP30, NP60) . In protocol 2, the following four groups of rats were studied : IP30, PP30, IP30+ glibenclamide group (G-IP30) and PP 30+G (G-PP3O) group. Hemodynamics and arrhythmia were observed continuously, and infarcted size and Ca⁺⁺-ATPase activity of sarcoplasmic reticulum (SR) in the Ischemic area were analyzed. There were no significant changes in hemodynamics. In protocol 1, the incidence of VT and infarcted size were significantly lower and Ca⁺⁺-ATPase activity of SR was significantly higher in groups IP30, IP60 and PP30 than those of groups NP30, NP60 and PP60. In protocol 2, glibenclamide pretreatment abolished the effects of preconditioning on these parameters in both the PP and IP groups. We conclude that the KATP channel may play an important role in the rat model of preconditioning.

View full abstract

Bone mineral density ( BMD ) decreases in Graves'disease due to the high rate of bone turnover with a predominance of resorption. Studies have evaluated the use of dual energy X-ray absorptiometry (DEXA) for investigating bone metabolism. Bone strength is a function of BMD and bone quality. Quantitative ultrasound (QUS), which involves no radiation exposure and uses portable equipment, assesses both BMD and bone quality. This study investigated the relationship between broadband ultrasound attenuation ( BUA ), speed of sound (SOS), and stiffness to BMD measured at the calcaneus in a closely matched region of interest (ROI) ; and the relationship between BMD and ultrasound parameters to markers of bone metabolism in female patients with Graves' disease. We investigated 9 patients with untreated Graves' disease (group G : mean age, 41.0±7.6 years) and 19 patients in remission from Graves' disease (group RG : mean age, 55.7±5.0 years) . Ultrasound parameters by QUS correlated significantly with calcaneus BMD in the ROI only for group RG, suggesting that bone quality played an additional role. In 8 patients who were in remission for more than 5 years, the Z-scores for ultrasound parameters of the calcaneus assessed by QUS were in the negative range. No significant correlation was found between blood or urine markers and ultrasound parameters or calcaneus BMD. These results show that the lack of a significant correlation between ultrasound parameters and BMD of the calcaneus in our present group G indicates that BUA and SOS are affected by mineral content and by other material and structural properties. In addition, even Graves' disease patients who are in long-term remission may not have had recovery of bone density to normal levels following hyperthyroidism.

View full abstract

In pancreatic acinar cells, Ca²⁺/calmodulin-dependent protein kinase (CaM-kinase) type II is activated by cholecystokinin (CCK) -8 in a dosedependent manner, whereas amylase secretion is inhibited by supramaximal doses of CCK-8. Bradykinin appears to be involved in exacerbation of caerulein-induced pancreatitis. In endothelial cells, filamin translocation, which is responsible for bradykinin release in the early inflammatory phase, is initiated by CaM-kinase II. Activation of CaM-kinase II in pancreatic acinar cells is therefore potentially involved in caerulein-induced pancreatitis. The aim of this study was to investigate whether CaM-kinase II plays an important role in caerulein-induced pancreatitis. Rats were given 4 intraperitoneal (i.p.) injections of caerulein. CaM-kinase II activity was measured by the rate of incorporation of [γ-³²P] ATP with autocamtide II as a specific substrate. CaM-kinase II was also detected by immunohistochemistry. At 6 hours after the initial i.p. injection of caerulein, the maximal pancreatic damage was histologically observed and maximal pancreatic tissue weight and minimal pancreatic protein content were found. Serum amylase levels were significantly elevated from 6 to 9 hours. The maximal CaM-kinase II activity was observed and acinar cells were intensely immunoreactive for CaM-kinase II at 6 hours. KN-62, a selective CaM-kinase II inhibitor, had a preventive effect on edema and vacuolization and significantly reduced serum amylase levels during pancreatitis. This study indicates that activation of CaM-kinase II in pancreatic acinar cells plays a crucial role in the early phase of caerulein-induced pancreatitis.

View full abstract

Cholecystokinins (CCKs) in extracts of human bile of patients with obstructive jaundice were compared with those in bile of patients without obstructive jaundice. Cholecystokinins were purified from human bile using chloroform with a Sephadex G-25 column and a Sep-Pak C₁₈cartridge and isolated by high performance liquid chromatography (HPLC) . The different molecular forms of cholecystokinin ( CCK ), especially cholecystokinin tetrapep-tide ( CCK-4), cholecystokinin octapeptide (CCK-8) and cholecystokinin triacontatriapeptide (CCK-33) in human bile were isolated by HPLC and measured by radioimmunoassay and bioassay. The concentrations of CCK measured by radioimmunoassay (RIA) were consistent with those by bioassay, indicating that bioactivity of CCK is not attenuated in human bile. Moreover, the CCK receptor antagonist L-364, 718 (10 nM) completely inhibited the bioactivity of CCK in human bile. We found that the predominant molecular forms of the CCK in human bile of patients with obstructive jaundice are larger than those in bile of patients without obstructive jaundice. These results suggested that the liver may be the major site of CCK extraction or degradation and may therefore play a significant role in CCK turnover.

View full abstract

This study retrospectively examined prognostic factors for acute myocardial infarction in 150 patients with cardiogenic shock among 1, 200 patients who developed acute myocardial infarction. The mortality was 35.3%. A history of old myocardial infarction, multi-vessel coronary lesions, unsuccessful reperfusion therapy, and infarction of the left main coronary trunk significantly influenced prognosis. In patients in whom circulatory reconstruc-tion led to discharge without residual ischemia, the prognosis was similar to that in patients with usual myocardial infarction. The prognosis of acute myocardial infarction with cardiogenic shock may be improved by performing reperfusion therapy earlier. If necessary, residual ischemia should be treated by additional circulatory reconstruction.

View full abstract

We examined the effects of antiprogesterone steroid RU486 (17β-hy-droxy-11 β- [4-dimethylamino-phenyl] -17α- [1 propynyl] -estra-4, 9-diene-3-one) on human luteal function in vitro. Corpus luteal tissues obtained at gynecological surgery with informed consent and were homogenized and incubated with labeled pregnenolone or testosterone as substrates. The effects of RU486 on 3β-hydroxysteroid dehydrogenase (3β-HSD) activity assessed by the formation of progesterone from pregnenolone, and aromatase activity measured by the conversion of testosteone to estrone and estradiol were examined. Minced corpus luteal tissues were cultured with Medium 199 in a gas phase of O₂: CO₂at a ratio of 95 : 5 for 1 hour with or without RU486. The concentra-tions of unconjugated- and conjugated pregnenolone, and progesterone from the tissues and media were measured by RIA. RU486 had no effect on 3β-HSD or aromatase activities when 800×g supernatant from corpus luteal tissue was incubated with labeled substrates. When minced tissuee was incubated with RU486, the formation of pregnenolone and progesterone was significantly decreased in both the medium and tissues. These results demonstrated for the first time that the suppression of pregnenolone production by RU486 might be partially due to the inhibition of de novo pregnenolone production in the steroid synthesis in the corpus luteum.

View full abstract

The primary purpose of this investigation was to elucidate the correlation between changes of thyroid function and the response to antidepressant drugs in patients experiencing a depressive episode. The study sample consisted of 81 patients between the ages of 18 and 85 years with a definite diagnosis of a depressive episode of a major depressive disorder or bipolar disorder (DSM-IV) . We administered pharmacotherapy with antidepressant drugs to the patients for 12 weeks. At the commencement of pharmacotherapy, and 6 and 12 weeks thereafter, we assessed the patients with the 17-item Hamilton Rating Scale for Depression (HRSD-17) and thyroid function tests (free T3, free T4 and TSH) . Subjects showing at least 50% improvement in the HRSD-17 score were defined as responders. Of the 50 subjects who could be followed up, 29 were responders (58.0%) . The serum free T4 levels (baseline) were significantly higher in the responders (1.42 ± 0.23) than in the nonresponders (1.24 ± 0.21) . Repeated-measure ANOVA yielded no significant main effect for any group (df=1, F=2.07, p=0.16), significant effect of time (df=1.76, F=25.2, p<0.001) or significant interaction of group X time (df=1.76, F= 6.43, p=0.004) for free T4. In 25 responders, the degree of decrease in the serum free T4 levels during the first term (from baseline to 6 weeks) was greater than that during the second term (from 6 weeks to 12 weeks) . Serum free T4 levels may be related to the response to antidepressant drugs and the clinical course of patients with depression.

View full abstract

We performed two independent evaluations of in situ polymerase chain reaction (in situ PCR) and Epstein-Barr virus encoded small RNA in situ hybridization (EBER ISH) for detection of Epstein-Barr virus infection in gastric carcinoma. In situ PCR is a novel technique with a high sensitivity which is a powerful aid for morphological diagnosis. Paraffin sections of gastric carcinomas from 96 cases were analyzed. Twenty-two of 96 cases (23%) were positive by in situ PCR, and 14 cases (15%) were positive by EBER ISH. Therefore there were 8 cases of EBV-DNA-positive gastric carcinoma without expression of EBER. To determine the significance of the expression of EBER in EBV-infection positive gastric carcinoma, we performed a clinicopathological study. There were two pathologic features in the gastric carcinoma with EBER. There was a significantly higher incidence of undifferentiated type and advanced cancer in EBER-positive gastric carcinomas. Therefore the expression of EBER contributes to the clinical pathologic feature of Epstein-Barr virus-associated gastric carcinoma.

View full abstract