A possible involvement of oxygen free radicals in the development of myocardial acidosis during coronary occlusion in dogs

Summary The present study was designed to examine whether free radical scavengers attenuate myocardial acidosis induced by partial occlusion of the coronary artery in dogs. The myocardial pH was determined by a micro glass pH electrode inserted in the endocardial layers of the left ventricular wall perfused by the left anterior descending coronary artery. The left anterior'descending coronary artery was occluded for 90 min incompletely so that the flow would be 1/2–1/3 the original flow. The myocardial pH before partial occlusion was 7.54–7.55. Partial occlusion decreased the flow in the left anterior descending coronary artery by 49.3–64.9% and the myocardial pH by 0.71–0.76, and increased the ST segment (surface electrocardiogram) by 6.3–9.3 mV. Saline (0.5 ml/kg), recombinant human superoxide dismutase (70,000 or 210,000 U/kg), or catalase (55,000 or 165,000 U/kg) was injected intravenously 30 min after partial occlusion. The injection of recombinant human superoxide dismutase or catalase alone did not restore the myocardial pH that had been decreased by coronary occlusion. The combined injection of recombinant human superoxide dismutase (70,000 U/kg) + catalase (55,000 U/kg), however, restored the myocardial pH without restoration of ST segment. In conclusion, recombinant human superoxide dismutase + catalase attenuated myocardial acidosis during ischaemia, suggesting a possible involvement of oxygen free radicals in the development of myocardial acidosis (especially in the endocardial layers) during ischaemia. Send offprint requests to Y. Abiko at the above adress     

The natural killer T (NKT) cell ligand alpha-galactosylceramide demonstrates its immunopotentiating effect by inducing interleukin (IL)-12 production by dendritic cells and IL-12 receptor expression on NKT cells

The natural killer T (NKT) cell ligand alpha-galactosylceramide (alpha-GalCer) exhibits profound antitumor activities in vivo that resemble interleukin (IL)-12-mediated antitumor activities. Because of these similarities between the activities of alpha-GalCer and IL-12, we investigated the involvement of IL-12 in the activation of NKT cells by alpha-GalCer. We first established, using purified subsets of various lymphocyte populations, that alpha-GalCer selectively activates NKT cells for production of interferon (IFN)-gamma. Production of IFN-gamma by NKT cells in response to alpha-GalCer required IL-12 produced by dendritic cells (DCs) and direct contact between NKT cells and DCs through CD40/CD40 ligand interactions. Moreover, alpha-GalCer strongly induced the expression of IL-12 receptor on NKT cells from wild-type but not CD1(-/-) or Valpha14(-/-) mice. This effect of alpha-GalCer required the production of IFN-gamma by NKT cells and production of IL-12 by DCs. Finally, we showed that treatment of mice with suboptimal doses of alpha-GalCer together with suboptimal doses of IL-12 resulted in strongly enhanced natural killing activity and IFN-gamma production. Collectively, these findings indicate an important role for DC-produced IL-12 in the activation of NKT cells by alpha-GalCer and suggest that NKT cells may be able to condition DCs for subsequent immune responses. Our results also suggest a novel approach for immunotherapy of cancer.     

Terminal restriction fragment length polymorphism analysis of the diversity of fecal microbiota in patients with ulcerative colitis

BACKGROUND: Terminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful tool to assess the diversity of complexed microbiota. This permits rapid comparison of microbiota from many samples. In this study, we performed T-RFLP analysis of the fecal microbiota from patients with ulcerative colitis (UC). METHODS: Forty-four patients with UC (23 women and 21 men, median age 25 years) and 46 healthy individuals (25 women and 21 men, median age 34 years) were enrolled in this study. DNA was extracted from their stool samples, and the 16S rRNA genes were amplified by PCR. The PCR products were then digested with HhaI and/or MspI restriction enzymes, and the length of the T-RF was determined. RESULTS: The fecal microbial communities were classified in 8 clusters. Almost all the healthy individuals (39 of 46) were included in cluster I, and most of the UC patients could be divided into the other 7 clusters, indicating that fecal bacterial communities are different between healthy individuals and active UC patients. Some T-RFs, derived from the unclassified bacteria, Ruminococcus, Eubacterium, Fusobacterium, gammaproteobacteria, unclassified Bacteroides, and unclassified Lactobacillus, were detected in the UC patients, but not in the healthy individuals. The T-RFLP patterns were also different between the active patients and inactive (remission) patients. The T-RF derived from the unclassified bacteria, Ruminococcus and Eubacterium, and the T-RFs derived from the unclassified bacteria, Eubacterium, and Fusobacterium were predominantly detected in the active patients not the inactive patients. In contrast, the T-RFs derived from Lactobacillus and unclassified Lactobacillus were more predominant in the inactive (remission) patients. In 4 patients with proctitis, the pattern of fecal microbial diversity was very similar. CONCLUSIONS: T-RFLP analyses showed that the diversity of fecal microbiota in patients with UC was different from that in healthy individuals. Unclassified bacteria, as well as known bacteria, can contribute to alterations in the bacterial diversity of UC patients.     

Immunological tolerance-related genes in a spontaneous tolerant model of rat liver transplantation explored by suppression subtractive hybridization

Natural immunological tolerance can be induced in certain types of allogeneic liver transplantation in rats. To screen for genes associated with the induction of tolerance, suppression subtractive hybridization was performed in the rat liver transplantation model between a DA donor and PVG recipient combination where spontaneous immunological tolerance is known to occur without any immunosuppressive treatment. As a result, 112 genes were cloned from a DA liver graft that survived for 20 days in the fully allogeneic PVG recipient. After confirmation of the expression intensity using an in-house manufactured DNA array with cDNAs from the DA graft, 36 genes were classified in the highly expressed group and 26 moderately expressed group. In the first group, there were 8 immunoglobulin-related genes and 6 MHC class II-related genes, suggesting the existence of an underlying rejection response. Among those genes, an antiapoptotic gene in the p38 MAP kinase pathway, heme oxygenase gene (HO-1), and a ras cascade gene, IQ motif containing GTPase activating protein 1 (Iqgapl), retained biological significance. The results suggested that the molecular response to a liver graft tends to be antiapoptotic and to terminate the rejection response. Unfortunately, there was no gene identified that qualified as a putative immunosuppressive protein, liver suppressor factor-1 (LSF-1). The panel of genes identified in the present work will be a useful panel of candidate genes to investigate the induction of spontaneous tolerance.     

Renal pathology of ANCA-related vasculitis: proposal for standardization of pathological diagnosis in Japan

BACKGROUND: In Japan, systematic evaluation of the histologic parameters of anti-neutrophil cytoplasmic autoantibodies (ANCA)-related vasculitis has been performed according to the Japanese classification by Shigematsu et al. However, this classification is quite different from that of the European Vasculitis Study Group (EUVAS) classification. Therefore, a histological common basis is needed to compare Japanese histological data with the international database. METHOD: Histological parameters concerning glomerular, tubulointerstitial, and vascular lesions of ANCA-related vasculitis, which are indispensable for clinical management, were elucidated and defined by reviewing, utilizing the merits of, and amending the two scoring systems. RESULTS AND CONCLUSION: A new comprehensive and standardized scoring system, by which histological quantitative assessment can provide evidence for therapy planning, has been developed for renal biopsy of Japanese ANCA-related vasculitis.     

The aging of accommodative function and its pupillary response in VDT workers without subjective complaints]

Using a computer-assisted infrared optometer with a pupillometer, we measured the accommodative function and its pupillary response in 81 female subjects (age: 31-55 yrs) without visual complaints who worked via visual-display-terminal (VDT). From these measurements we analyzed the following 6 parameters regarding accommodation, and investigated whether a correlation exists between age and each parameter. The six parameters were, (1) the amplitude of objective accommodation, (2) the refractive fluctuation at the resting state of accommodation, (3) the refractive fluctuation after receiving myopic stimuli, (4) the pupil area prior to myopic stimuli, (5) the maximum rate of constriction by accommodation, and (6) the rate of pupil area recovery after accommodation. Among these parameters, the amplitude of objective accommodation, the refractive fluctuation at the resting state of accommodation, the refractive fluctuation after receiving myopic stimuli, and the pupil area prior to myopic stimuli were significantly correlated with age (all parameters: p less than 0.001). The correlation coefficients (r) were -0.810, -0.591, -0.612 and -0.320 respectively. There was also some relationship between age and the rate of pupil area recovery after accommodation (r = 0.188, p = 0.018), but no statistically significant correlation between age and the rate of constriction by accommodation (r = 0.100). These findings suggest that the overall functional capability of accommodation decreases constantly with age, though the pupillary reaction induced by accommodation is not correlated well with age.     

Recurrent thymoma in patients with myasthenia gravis

One hundred sixty-six patients underwent operation for myasthenia gravis between 1977 and 1989. Thirty-eight patients had associated thymoma, registering stages I (n = 17), II (n = 9), III (n = 11), and IVa (n = 1) according to the classification of Masaoka and colleagues. Extended thymectomy was performed on 128 patients without thymoma; thymothymectomy, with resection of the anterior mediastinal fat and tissues adherent to the tumor, was performed in all patients with thymoma. There were no instances of early or late death. Neuromuscular function improved, and clinical myasthenic symptoms stabilized in almost all patients except 2 patients in stage III and 1 patient in stage IVa, who had an exacerbation of the myasthenic symptoms associated with recurrence of thymoma. All the recurrent tumors were on the pleura and could be resected. The suspected cause of recurrence is either dissemination of tumor cells as a result of operative manipulation or undetected disseminated foci that existed at the time of the first operation. The resections of the recurrent invasive thymomas localized on the pleura were easily performed and improved the myasthenic symptoms.     

Value of thyroid stimulating antibody in the diagnosis of thyroid associated ophthalmopathy of euthyroid patients

AIMS/BACKGROUND—Thyroid associated ophthalmopathy (TAO) of euthyroid patients is difficult to diagnose because clinical findings overlap with other conditions, and no confirmatory diagnostic tests are available. Recently, it was reported that TSH binding inhibitor immunoglobulin (TBII) and thyroid stimulating antibody (TSAb) are sensitive markers of TAO. The sensitivity of these antibodies in the detection of TAO were therefore studied to determine if they could be a useful criterion in the diagnosis of TAO of euthyroid patients.
METHODS—Serum values of TBII and TSAb of 35 patients with euthyroid TAO (group A) were compared with those of 27 patients with Graves' disease and TAO (group B). The relation between the serum value of TSAb and the eye symptoms of patients with euthyroid TAO were also examined by multiple linear regression analysis.
RESULTS—In group A, TBII was positive in 10 cases (28.6%) and TSAb was positive in 29 cases (82.9%). In group B, both TBII and TSAb were positive in all cases (100%). The titre of serum TBII in group A (15.6% (SD 18.0%)) was significantly lower (p<0.0001) than in group B (57.9% (21.5%)). The titre of serum TSAb in group A (1400.9% (2163.9%)) was significantly lower (p=0.0026) than in group B (2243.9% (1472.8%)). Among the eye findings of patients with euthyroid TAO, keratopathy was significantly (p=0.034) related to the value of TSAb.
CONCLUSION—These results suggest that the activity of TSAb is a more sensitive marker of euthyroid TAO than is TBII, and could be a useful criterion in the diagnosis of TAO of euthyroid patients.