Phenotypic and genetic analyses of T-cell-mediated immunoregulation in patients with Type 1 diabetes.

AIMS: To investigate the contribution of regulatory T cells and co-stimulatory molecules in CD4(+) T cells to the development of Type 1 diabetes (T1D). METHODS: Twelve patients with T1D, nine patients with systemic lupus erythematosus (SLE), and 12 age-matched healthy control subjects participated. We analysed the proportions of CD25(+)CD4(+) T cells and natural killer T cells (NKT cells), and the expression levels of Foxp3, CTLA-4, CD28, ICOS, PD-1 and BTLA in peripheral blood mononuclear cells and purified CD4(+) T cells. RESULTS: There were no significant differences in the proportions of CD25(+) CD4(+) T cells or NKT cells among the three groups. PD-1 expression levels of peripheral CD4(+) T cells from T1D patients were significantly lower than those from healthy control subjects (P = 0.00066). In contrast, PD-1 expression levels were similar in SLE patients and healthy control subjects. The expression levels of Foxp3, CTLA-4, CD28, ICOS and BTLA were similar in the three groups. CONCLUSIONS: Decreased expression of the PD-1 gene in CD4(+) T cells may contribute to the development and/or maintenance of autoimmune T1D. As the population studied was small and heterogeneous, further studies are required to confirm the findings.

Cytogenetic analysis of myxoid liposarcoma and myxofibrosarcoma by array-based comparative genomic hybridisation.

AIM: To investigate overall chromosomal alterations using array-based comparative genomic hybridisation (CGH) of myxoid liposarcomas (MLSs) and myxofibrosarcomas (MFSs). MATERIALS AND METHODS: Genomic DNA extracted from fresh-frozen tumour tissues was labelled with fluorochromes and then hybridised on to an array consisting of 1440 bacterial artificial chromosome clones representing regions throughout the entire human genome important in cytogenetics and oncology. RESULTS: DNA copy number aberrations (CNAs) were found in all the 8 MFSs, but no alterations were found in 7 (70%) of 10 MLSs. In MFSs, the most frequent CNAs were gains at 7p21.1-p22.1 and 12q15-q21.1 and a loss at 13q14.3-q34. The second most frequent CNAs were gains at 7q33-q35, 9q22.31-q22.33, 12p13.32-pter, 17q22-q23, Xp11.2 and Xq12 and losses at 10p13-p14, 10q25, 11p11-p14, 11q23.3-q25, 20p11-p12 and 21q22.13-q22.2, which were detected in 38% of the MFSs examined. In MLSs, only a few CNAs were found in two sarcomas with gains at 8p21.2-p23.3, 8q11.22-q12.2 and 8q23.1-q24.3, and in one with gains at 5p13.2-p14.3 and 5q11.2-5q35.2 and a loss at 21q22.2-qter. CONCLUSIONS: MFS has more frequent and diverse CNAs than MLS, which reinforces the hypothesis that MFS is genetically different from MLS. Out-array CGH analysis may also provide several entry points for the identification of candidate genes associated with oncogenesis and progression in MFS.

[Changes in serum malondialdehyde-modified low-density lipoprotein in patients with acute myocardial infarction and stable angina pectoris treated by coronary angioplasty].

OBJECTIVES: Serum malondialdehyde-modified low-density lipoprotein(MDA-LDL) was measured as a marker of oxidative stress, and the changes in serum MDA-LDL compared in patients with acute myocardial infarction and stable angina pectoris treated by percutaneous transluminal coronary angioplasty (PTCA). METHODS: Forty-one patients with acute myocardial infarction or stable angina pectoris were admitted to our hospitals between January 2000 and June 2000. Direct PTCA was performed in 17 patients(MI group) and elective PTCA in 24 patients(AP group). Coronary angiography was performed in nine control subjects(control group). Serum MDA-LDL was measured in the peripheral venous blood before and immediately after procedures in each group(normal range 20-80 U/l). RESULTS: There were no significant differences in patient characteristics, except age, between the MI group and AP group. Serum MDA-LDL was elevated above the normal range before the procedure in both groups(MI group 104.7 +/- 52.0 U/l, AP group 99.7 +/- 42.8 U/l), and significantly decreased immediately after the procedure(MI group 61.3 +/- 25.6 U/l, AP group 62.0 +/- 29.6 U/l), but there were no significant differences between the two groups. Serum MDA-LDL was elevated before the procedure (99.3 +/- 48.9 U/l) in the control group and significantly decreased immediately after the procedure(61.7 +/- 26.2 U/l). However, these values did not differ from the values before and immediately after the procedure in the MI group and the AP group. The percentage changes in serum MDA-LDL before and immediately after the procedure were -38 +/- 16% in the MI group, -37 +/- 17% in the AP group and -36 +/- 20% in the control group, and there were no significant differences between the three groups. CONCLUSIONS: No significant difference in the changes in serum MDA-LDL was observed between patients with acute myocardial infarction and stable angina pectoris treated by PTCA. However, anticoagulants may affect the MDA-LDL measurements directly, because similar changes in serum MDA-LDL were observed in control subjects after only coronary angiography.

Synchronous primary lung carcinoma and lung metastasis from extrathoracic carcinoma.

We present the cases of 2 patients in whom primary lung cancer was found unexpectedly when pulmonary resection was performed for metastatic lung cancer. The possibility of combined primary and metastatic carcinoma should be considered in patients with a diagnosis of multiple pulmonary metastases from extrathoracic tumor.

Association of CTLA-4 but not CD28 gene polymorphisms with systemic lupus erythematosus in the Japanese population.

OBJECTIVE: Systemic lupus erythematosus (SLE) in a multisystem autoimmune disorder characterized by multiorgan pathology and autoantibodies against a variety of autoantigens. The CD28 and CTLA-4 genes might be candidate genes for SLE, because costimulation signals from CD80/CD86 to CD28/CTLA-4 have been suggested to play an important role in the activation or inactivation of T lymphocytes. METHODS: We investigated three polymorphic regions within the CTLA-4 gene, a C/T base exchange in the promoter region -318 (CTLA-4 -318C/T), an A/G substitution in the exon 1 position 49 (CTLA-4 49A/G), an (AT)(n) repeat polymorphism in the 3' untranslated region of exon 4 [CTLA-4 3' (AT)(n)], and a CD28 gene polymorphism, a T/C substitution in the intron 3 position +17 (CD28 IVS3+17T/C), in SLE patients and controls. RESULTS: SLE patients had significantly higher frequencies of the CTLA-4 49G allele (P=0.003) and of the CTLA-4 (AT)(n) 106 bp allele (P=0.0008) than controls. We also found a strong linkage disequilibrium between the A allele of CTLA-4 49A/G and the 86 bp allele of CTLA-4 3' (AT)(n). On the contrary, no association was found between SLE and CTLA-4 -318C/T or CD28 IVS3 +17T/C. CONCLUSION: We conclude that the CTLA-4 gene appears to play a significant role in the development of SLE in the Japanese population.

[A case of surgically treated left atrial myxoma following acute multiple embolism including cerebral embolism].

The patient was a 59-year-old female who was admitted to the hospital due to acute pain of bilateral legs, a numbness of right hand and anarthria. Angiography of extremities revealed total occlusion of right ulnar artery, left radial artery and bilateral popliteal arteries. Brain MRI revealed multiple small infarctions. Echocardiography revealed a mass in the left atrium. She was diagnosed as multiple embolism including cerebral embolism caused by left atrial myxoma. Open heart surgery immediately after the attack is generally considered contraindicated due to problems of hemorrhagic infarction or brain edema. But, relapse of embolism may deteriorate the condition and miss the timing of surgery. Thus we performed removal of left atrial myxoma 4 days after the attack. The postoperative course was uneventful. This is a few report about open heart surgery immediately after the attack. We report about the indication and the optimal timing of open heart surgery following cerebral embolism.

Ultrafiltration of the priming blood before cardiopulmonary bypass attenuates inflammatory response and improves postoperative clinical course in pediatric patients.

The priming solution using in cardiopulmonary bypass (CPB) for infants undergoing cardiac surgery includes considerable amounts of stored blood. Our objective was to test the hypothesis that ultrafiltration (UF) of the stored blood before CPB reduces the unfavorable effects of stored blood and the production of inflammatory cytokines. Fifty pediatric patients with congenital heart defects took part in this study. The patients were randomly divided into two groups: the UF (27 pediatric patients who received UF) and control (23 pediatric patients who did not receive UF) groups. UF was performed with a polysulphone ultrafiltrator before CPB. Blood samples were collected immediately before, during, and 1 h after CPB. The levels of cytokines (TNF-alpha, IL-1beta, IL-8), NH3, and bradykinin were determined. The serum concentrations of NH3 and bradykinin decreased significantly after UF. Compared with the control group, the UF group had significantly lower cytokine production. Water balance in UF group was better than that of control group. The UF group received significantly less inotropic support and shorter duration of ventilator support and ICU stay. We conclude that removal of bradykinin and a decrease in the levels of NH3, potassium, and pH play a significant role in reducing water retention and postoperative lung injury. UF of the blood used to prime the circuit for CPB is a safe and efficient method for use in open heart surgery in small pediatric patients.

[The perioperative management for the patients to undergo open heart surgery with chronic renal failure].

From January, 1996 to December, 1999, eight patients with chronic renal failure received open heart surgery. They consists of six males and two females aged between 45 and 72 with a mean of 59.6. The duration of hemodialysis was 4.0 years in a mean. Seven of them had isolated coronary artery bypass grafting (CABG), one of them had CABG and aortic valve replacement. All patients were dialysed dialy two days before operation. Intraoperative hemodialysis (HD) was used in all patients. In recent six patients extracorporeal ultrafiltration methods (ECUM) were also performed intraoperatively in addition to HD. In postoperatively continuous hemodiafiltration (CHDF) has been preferred to HD in all patients, and nafamostat mesilate is a useful anticoagulation agent to prevent postoperative bleeding complications. The duration of CHDF was 3.2 days in a mean (the shortest for one day and the longest for eight days). When the circulatory situation were stable, HD was performed on early postoperative day. One patients died of low output syndrome and multiple organ failure. We reported problems in perioperative management of patients with chronic renal failure and our protocol.

Complement component 9 deficiency is not a susceptibility factor for SLE.

Almost all complement component 9 (C9) deficiency in Japan shows Arg95 Stop mutation of C9 gene. Therefore, we studied the prevalence of Arg95Stop mutation of C9 gene among 78 patients with SLE to elucidate the association of SLE and C9 deficiency. The Arg95Stop carrier frequency showed no significant difference between SLE patients and controls. Thus, C9 deficiency is not implicated in SLE susceptibility.

Atopic dermatitis as a risk factor for acute native valve endocarditis.

Colonization of Staphylococcus aureus is commonly observed in skin lesions of atopic dermatitis (AD) patients, and scratching of the pruritic lesions may lead to reiterative bacteremia. It is possible that acute native valve endocarditis may develop in a patient with uncontrolled AD; the latter condition may be a risk factor for the former. We report two cases of acute aortic and/or mitral valve endocarditis complicated with recurrent cutaneous infections caused by severe AD. The patients underwent successful surgical treatment of the heart lesions, plus intensive postoperative antibiotics and skin treatment for AD.

Biocompatibility of silicone-coated oxygenator in cardiopulmonary bypass.

BACKGROUND: This study was designed to analyze the biocompatibility of silicone-coated oxygenators using inflammatory response as the outcome measure, and to investigate whether the silicone-coated oxygenators perform better in terms of postoperative organ dysfunction. METHODS: The 32 patients who underwent cardiopulmonary bypass (CPB) were divided into 3 groups: group A (n = 10), heparin-coated circuit with silicone-coated oxygenator; group B (n = 11), whole heparin-coated circuit; and group C (n = 11), whole untreated circuit. The plasma concentrations of the proinflammatory markers, made of inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, interleukin-8), terminal complement complex (C5b-9), and polymorphonuclear elastase (PMN-E), were measured by enzyme-linked immunosorbant assay. RESULTS: All proinflammatory markers were significantly lower in groups A and B than in group C, especially C5b-9 and PMN-E concentrations, which were significantly lower in group A than in group B. The alveolar-arterial oxygen gradients (A-aDO2) and the respiratory index were significantly better in group A than in group C. In group B, however, only the A-aDO2 was significantly better than in group C. The duration of intubation and the length of stay in the intensive care unit stay were significantly shorter in groups A and B than in group C. CONCLUSIONS: Silicone-coated oxygenators are biocompatible and prevent postoperative organ dysfunction.

Molecular epidemiology of C9 deficiency heterozygotes with an Arg95Stop mutation of the C9 gene in Japan.

Deficiency of the ninth component of human complement (C9) is the most common complement deficiency in Japan, with an incidence of approximately one homozygote in 1000, but is very rare in other countries. Genetic analyses of Japanese C9 deficiency have shown that a C-to-T transition leading to TGA stop codon for Arg95 in exon 4 of the C9 gene (Arg95Stop) is common in Japanese C9 deficiency. To determine the prevalence of heterozygous carriers of the Arg95Stop mutation in a Japanese population, we collected DNA samples from 300 individuals in two of the four main islands of Japan. Heterozygote detection was performed with an allele-specific polymerase chain reaction (PCR) system designed to detect exclusively only one of the normal and mutant alleles, followed by confirmation with PCR/single-strand conformation polymorphism (SSCP) analysis and direct sequencing. Twenty individuals were heterozygous for the Arg95Stop mutation. None was homozygous. The prevalence of carriers of the Arg95Stop mutation was 6.7% (20/300). An estimated frequency (0.12%) of complete C9 deficiency due to homozygous Arg95Stop mutation was consistent with frequencies determined by serological studies.

[Completion pneumonectomy combined with graft replacement of thoracic aortic aneurysm by simple clamping].

A 59-years-old male patient who had left upper lobe partial resection 30 years ago. He was seen at the family physician because of cough. A chest X-ray was showing an abnormal mass shadow measuring 3 x 4 cm in left lower lobe like honey comb. And squamous cell carcinoma (SCC) was detected in his sputum. He was diagnosed as primary lung cancer and introduced to our department to have operation. Chest CT-scan was showing lung tumor suspected SCC measuring 4.3 x 2.6 cm in segment 8 faced chest wall. At the same time, we detected thoracic aortic aneurysm and subcarinal lymph node, but could not see where the boundary is, so it was hard to distinguish between parietal thrombus with thoracic aortic aneurysm and swelling subcarinal lymph node. We decided it swelling subcarinal lymph node by three-dimensional treated CT-scan. Aortic angiography was showing proximal descending aortic aneurysm measuring diameter was 4.5 cm. Abdominal CT-scan was showing infrarenal abdominal aortic aneurysm measuring diameter was 5.5 cm. He was diagnosed as primary lung cancer (It. S8, SCC) (cT2N2M0, Stage IIIB), thoracic aortic aneurysm, abdominal aortic aneurysm, and idiopathic pulmonary fibrosis, and had completion pneumonectomy (R 2 b) for primary lung cancer and graft replacement with aneurysm dissection for thoracic aortic aneurysm without extracorporeal circulation. In this operation, we could find swelling subcarinal lymph node measuring 5 x 3 cm instead of parietal thrombus with thoracic aortic aneurysm. Pathological examination diagnosed middle differential SCC and no metastasis from dissected lymph node (PT2N0M0, Stage I A).

Short-term and long-term effects of low-density lipoprotein (LDL) apheresis on restenosis after percutaneous transluminal coronary angioplasty (PTCA): is lowering Lp(a) by LDL apheresis effective on restenosis after PTCA?

It has been reported that serum lipoprotein(a) (Lp[a]) levels in patients with restenosis after percutaneous transluminal coronary angioplasty (PTCA) were significantly higher than in patients without restenosis. In this study, we evaluated the preventive effect of LDL apheresis on restenosis after PTCA in patients with hypercholesterolemia. For 10 patients who had shown a serum cholesterol level of more than 220 mg/dl despite treatment with antihypercholesterolemic drugs, LDL apheresis was conducted every 2 weeks after a successful PTCA until restenosis could be checked. In 4 patients, LDL apheresis was conducted for 2 years. LDL apheresis significantly reduced serum cholesterol from 248 +/- 22 mg/dl to 135 +/- 26 mg/dl and Lp(a) from 42 +/- 34 mg/dl to 21 +/- 16 mg/dl. The average degree of stenosis in the 11 lesions undergoing PTCA was 92 +/- 6% before PTCA, 35 +/- 10% immediately after PTCA, and 38 +/- 19% at 3 to 4 months after PTCA. Restenosis was observed in only 1 lesion. In 4 patients who received LDL apheresis for 2 years, restenosis did not occur in any of the 4 lesions treated. We concluded that LDL apheresis was an efficacious therapy to prevent restenosis after PTCA in patients with hypercholesterolemia.

Transcapillary ultrafiltration and peritoneal equilibration test in pediatric patients.

The usefulness of the peritoneal equilibration test (PET) in children is unknown. The relationship between transcapillary ultrafiltration and PET was investigated in order to evaluate the usefulness of PET in children. PET was performed on 14 patients undergoing peritoneal dialysis. Their age and bodyweight ranged from 3.8 to 23.6 years and 10.2 to 55.8 kg, respectively. The patients were divided into two groups according to bodyweight; group A patients weighed < or = 40 kg (n = 7) and group B patients weighed > 40 kg (n = 7). There was no significant difference in the mean infusion volume per bodyweight between the two groups, but the mean infusion volume per body surface area was smaller in group A than in group B. Group A showed a more rapid equilibration of dialysate glucose and creatinine than group B. Higher normalized mass transfer area coefficients were evident in group A. In spite of the lower effective glucose gradient in group A, the transcapillary ultrafiltration capacity (TUFC) showed no difference between the two groups. When the slope indices of the regression equations between the two groups were compared, the slopes of the regression in the relationship between TUFC and dialysate (D) ratios D/Do glucose or D/P creatinine in group A were steeper than those in group B. Results of the present study indicate that the larger peritoneal area to infusion volume in patients with smaller body size results in both a rapid equilibration of solutes and sufficient transcapillary ultrafiltration.

Effects of LDL apheresis on restenosis after angioplasty.

We performed long-term maintenance LDL apheresis therapy on patients with hypercholesterolemia after undergoing PTCA, and investigated the therapeutic effects of reducing serum cholesterol, LDL-cholesterol and Lp(a). LDL apheresis significantly reduced serum lipids and was an efficacious therapy in the prevention of recurrent stenosis after PTCA.

Disappearance of mitral valve regurgitation after successful percutaneous transluminal coronary angioplasty.

Percutaneous transluminal coronary angioplasty has been reported to improve several clinical parameters. Functional papillary muscle dysfunction, which is also known to induce mitral valve regurgitation, is reversible after revascularization. We described a patient, with a 95% stenosis of proximal right coronary artery, whose mitral valve regurgitation disappeared after successful percutaneous transluminal coronary angioplasty.

Oral mucositis and salivary methotrexate concentration in intermediate-dose methotrexate therapy for children with acute lymphoblastic leukemia.

The relationship between salivary methotrexate (MTX) concentration and severity of oral mucositis after administration of MTX was investigated in six children with acute lymphoblastic leukemia. They received two administrations of MTX at 500 mg/m2 with one third given bolusly and the remainder by 24-hour continuous infusion. No significant difference among patients or administration session was observed in serum MTX concentration. Detectable concentrations of salivary MTX (greater than 0.01 microM) were observed during nine of the ten infusions. A concentration of 0.1 microM or more, apparently lasting at least 12 hours, was observed during one infusion and followed by severe mucositis. During two of the ten infusions for different patients, concentrations of 0.04 to 0.07 microM and 0.02 to 0.04 microM, apparently lasting at least 12 and 18 hours, respectively, were observed, followed by moderate mucositis. During the other seven infusions, either much shorter or no increase in salivary MTX concentration was observed, with only mild or no subsequent mucositis. Analysis by Kendall's rank method showed a statistical correlation between concentration at 6 hours of infusion and severity of oral mucositis. The findings suggest that the early secretion of MTX into saliva has a significant role in the development of oral mucositis in leukemic children.