Association study of genetic variants on chromosome 7q31 with susceptibility to normal tension glaucoma in a Japanese population.

The caveolin 1 to caveolin 2 (CAV1-CAV2) gene region on chromosome 7q31 has been reported to be associated with susceptibility to primary open angle glaucoma (POAG) and normal tension glaucoma (NTG) in previous studies. We investigated whether genetic variants in the CAV1-CAV2 region are associated with NTG in Japanese patients. Two hundred and ninety-two Japanese patients with NTG and 352 Japanese healthy controls were recruited. We genotyped three single-nucleotide polymorphisms; that is, rs1052990, rs4236601, and rs7795356, in the CAV1-CAV2 gene region and assessed the allelic diversity among cases and controls. The frequency of the minor allele (G) of rs1052990 was significantly decreased in NTG cases compared with controls (P=0.014, OR=0.71), whereas NTG or POAG cases had a significantly higher frequency of the allele than controls in previous studies. Conversely, rs7795356 did not show any significant association with NTG cases, and rs4236601 was monomorphic in the Japanese study population. Our findings did not correspond with previous positive results, suggesting that CAV1-CAV2 variants studied in the present study are not important risk factors for NTG susceptibility in all populations. Further studies are needed to elucidate the possible contribution of the CAV1-CAV2 region to the development of glaucoma.

Lack of IL-21 signal attenuates graft-versus-leukemia effect in the absence of CD8 T-cells.

Previously, we have shown that IL-21R(-/-) splenocytes ameliorate GVHD as compared with wild-type splenocytes. Here, we investigated whether or not IL-21R(-/-) splenocytes diminish the graft-versus-leukemia (GVL) effect. Surprisingly, IL-21R(-/-) splenocytes efficiently eliminate leukemic cells as well as wild-type splenocytes, suggesting the retention of GVL effects in the absence of IL-21 signaling. To compare the GVL effect between IL-21R(-/-) and wild-type cells, we titrated the number of splenocytes required for the elimination of leukemic cells and found that the threshold of GVL effect was obtained between 5 × 10(5) and 5 × 10(6) with both types of splenocytes. Cotransplantation with CD8-depleted splenocytes but not with purified CD8 T-cells resulted in a significant reduction in anti-leukemic effect of IL-21R(-/-) cells compared with wild-type cells, suggesting that the lack of IL-21 signaling primarily impairs CD4 T-cell rather than CD8 T-cell function and the comparable GVL effect with IL-21R(-/-) bulk splenocytes results from cooperative compensation by CD8 T-cells.

IL-21 is critical for GVHD in a mouse model.

Immunological effects of IL-21 on T, B and natural killer (NK) cells have been reported, but the role of IL-21 in GVHD remains obscure. Here, we demonstrate that morbidity and mortality of GVHD was significantly reduced after BMT with splenocytes from IL-21R(-/-) mice compared with those from wild type mice. To further confirm our observation, we generated a decoy receptor for IL-21. GVHD was again less severe in mice receiving BM cells transduced with the IL-21 decoy receptor than control mice These results suggest that IL-21 critically regulates GVHD, and that blockade of the IL-21 signal may represent a novel strategy for the prophylaxis for GVHD.

Investigation of the association between the GLC3A locus and normal tension glaucoma in Japanese patients by microsatellite analysis.

PURPOSE: To investigate whether the GLC3A locus harboring the CYP1B1 gene is associated with normal tension glaucoma (NTG) in Japanese patients. MATERIALS AND METHODS: One hundred forty-two Japanese patients with NTG and 101 Japanese healthy controls were recruited. Patients exhibiting a comparatively early onset were selected as this suggests that genetic factors may show stronger involvement. Genotyping and assessment of allelic diversity was performed on 13 highly polymorphic microsatellite markers in and around the GLC3A locus. RESULTS: There were decreased frequencies of the 444 allele of D2S0416i and the 258 allele of D2S0425i in cases compared to controls (P = 0.022 and P = 0.034, respectively). However, this statistical significance disappeared when corrected (Pc > 0.05). We did not find any significant association between the remaining 11 microsatellite markers, including D2S177, which may be associated with CYP1B1, and NTG (P > 0.05). CONCLUSIONS: Our study showed no association between the GLCA3 locus and NTG, suggesting that the CYP1B1 gene, which is reportedly involved in a range of glaucoma phenotypes, may not be an associated factor in the pathogenesis of NTG.

New susceptibility locus for high myopia is linked to the uromodulin-like 1 (UMODL1) gene region on chromosome 21q22.3.

PURPOSE: To ascertain and define the position of a potential disease susceptibility gene around D21S0083i prioritized during our previous whole genome case-control association analysis with 27,158 microsatellite markers, in Japanese high-myopia patients. METHODS: 520 high myopic patients and 520 healthy controls were genotyped using 39 SNPs distributed around D21S0083i on chromosome 21q22.3. RESULTS: Only 1 SNP (rs2839471) of 39 SNPs was significant after correction for multiple testing (allele T: P=0.00027, Pc=0.01, OR=1.684). The SNP (rs2839471) did not reside in haplotype blocks constructed by the pair-wise linkage disequilibrium between the SNPs. CONCLUSIONS: The SNP (rs2839471) is suggested to be located in the frequent recombinant region within UMODL1. Together this region might play a critical role for susceptibility to high myopia, and warrants further confirming studies and investigations as to the mechanisms by which UMODL1 may contribute to myopia.

Interferon-gamma and NF-kappaB mediate nitric oxide production by mesenchymal stromal cells.

Mesenchymal stromal cells (MSCs) have been shown to have an immunosuppressive effect. Previously, we demonstrated that nitric oxide (NO) is one of the immunomodulatory mediators of MSCs. We herein show that primary mouse bone marrow MSCs and three cell lines that mimic MSCs suppress both differentiation and proliferation in Th1 condition, whereas the suppression in Th2 condition is mild. NO production is inversely correlated with T cell proliferation in Th1 and Th2 conditions. NO is highly induced in Th1 and minimally induced in Th2. Moreover, an inhibitor of NO synthase restores both proliferation and interferon-gamma (IFN-gamma) production in Th1 condition. Furthermore, an anti-IFN-gamma antibody strongly inhibits NO production and an inhibitor of NF-kappaB reduces the level of induction of inducible NO synthase (iNOS) in MSCs. Taken together, our results suggest that NO plays a significant role in the modification of Th1 and Th2 differentiation by MSCs, and that both IFN-gamma and NF-kappaB are critical for NO production by MSCs.

Screening of genes responsible for differentiation of mouse mesenchymal stromal cells by DNA micro-array analysis of C3H10T1/2 and C3H10T1/2-derived cell lines.

BACKGROUND: The molecular mechanisms underlying the biologic effects or differentiation of mesenchymal stromal cells (MSC) have not been clarified. Screening for genes differentially expressed at different stages is an important step in determining these molecular mechanisms. METHODS: In this study, we analyzed the gene expression profiles of C3H10T1/2 (10T1/2) cells and two sublines, A54 (pre-adipocyte) and M1601 (myoblast), as a model of MSC and downstream committed progenitors. RESULTS: We found up-regulated expression of delta-like-1 (Dlk), Wnt-5a and IL-1 receptor-like-1 (ST2) in 10T1/2 cells; stem cell factor (SCF) and stromal derived factor-1 (SDF-1) in A54 cells; and cardiac muscle-specific gene in M1601 cells. Overexpression of Dlk in A54 cells did not induce any effects on their differentiation into adipocytes. After differentiation into adipocytes, A54 cells reduced the expression of SCF, SDF-1 and Ang-1 as well as the ability to support the formation of a cobblestone appearance. DISCUSSION: The results suggest that these three lines hae different gene profiles and are a useful system for analyzing the differentiation and function of MSC and progenitor cells.

Four-digit allele genotyping of the HLA-A and HLA-B genes in Japanese patients with Behcet's disease by a PCR-SSOP-Luminex method.

The present study represents the first four-digit allele genotyping of HLA-A and -B in Japanese Behcet's disease (BD) patients and controls using a new genotyping method (named the PCR-SSOP-Luminex method) to determine the association of certain HLA-A or -B alleles with BD. Peripheral blood lymphocytes were collected from 180 Japanese BD patients and 170 healthy controls. The genotype frequency of HLA-B*5101 was significantly increased in the patients (61.7%) as compared with the controls (15.9%) (Pc = 1 x 10(-16), OR = 8.5). When we recalculated the phenotype frequencies after excluding the HLA-B*51-positive patients and controls to account for the effects of the linkage disequilibrium and the abundance of the HLA-B*51 allele, the frequencies of HLA-A*2602 and HLA-B*3901 had a weak association in the patient group without HLA-B*51 as compared with the control group without HLA-B*51 (A*2602; Pc = 0.130, OR = 4.3, B*3901; Pc = 0.099, OR = 3.5). This study confirmed on the basis of using a new and more accurate genotyping method that Japanese BD patients have a strong primary association with HLA-B*5101. The significant increase of HLA-A*2602 and B*3901 in the patient group without HLA-B*51 suggests that these two alleles might also have some secondary influence on the onset of BD.

[Preoperative magnetic resonance imaging evaluation of the aortic annular diameter in aortic valve stenosis].

Accurate preoperative knowledge of the aortic annular diameter is a great value in determining the appropriate procedure for aortic valve stenosis. We have successfully used magnetic resonance imaging (MRI) to evaluate the size of the aortic annular diameter preoperatively. Between October 1997 and October 1998, 5 aortic valve replacements were performed in patients with aortic valve stenosis. MRI was performed preoperatively in all cases. All images were gated to the R-wave of the electrocardiogram. Images representing 5 mm slices were acquired in the plane which is parallel to the line through the center points of the aortic root and left ventricle outflow tract. The maximum diameter of the aortic annulus (D) was measured at the point just below the attachment of the aortic valve leaflets. This diameter was compared to the maximum diameter of the prosthetic valve sizer (D 1) which could be easily inserted into the aortic annulus intraoperatively, and to the diameter of the prosthetic valve (D 2) that ultimately was implanted. Twenty-five minutes were required to scan each patient. There were no complications related to scanning. Mean values were D = 23.4 mm, D 1 = 22.3 mm and D 2 = 21.4 mm. There was a significant correlation between D and D 1, and D 2 was approximately 2 mm smaller than D. MRI evaluation is an accurate, noninvasive method for determining the aortic annular diameter preoperatively. We highly recommend this method for measuring the aortic annular diameter in case of aortic valve stenosis as part of the preoperative evaluation.

[A case of coronary artery bypass grafting using patch mounted graft technique for aortitis syndrome with left coronary ostial stenosis].

Coronary artery bypass grafting (CABG) was performed on a 36-year-old female for treatment of aortitis syndrome with left coronary ostial stenosis. The left anterior descending artery was anastomosed with the left internal thoracic artery. The circumflex artery was anastomosed with the saphenous vein. In the proximal anastomosis, the ascending aorta was sutured with a gelatin sealed knitted Dacron graft patch and anastomosed with the saphenous vein to prevent late occlusion due to intimal proliferation. Graft angiography 40 days and one year after the operation showed good patency of both grafts. Patch mounted graft technique seemed to be suitable for CABG in aortitis patients with the high incidence of late occlusion.

[Graft replacement for repair of coarctation of the aorta in adults: a case report].

A 42-year-old woman admitted to the hospital because of systolic murmur and upper body hypertension. Cardiac catheterization revealed tubular coarctation of the aorta and 60 mmHg pressure gradient across the coarctation. The graft replacement for repair of coarctation of the aorta was performed under femoro-femoral cardiopulmonary bypass. Since about 49% of surgically untreated patients of this disease may be expected to die before 30 years of age, repair of coarctation of the aorta in adults should be performed as soon as possible.

[Primary carcinosarcoma of TE lung--a report of two cases].

Carcinosarcoma is a rare lung tumor and accounts for less than 0.3% of primary lung malignancies. Since the first description by Kika in 1908, only 36 cases with this kind of tumor have been appeared in the Japanese literatures by 1993. This report presents our surgical experiences of two cases with carcinosarcoma of the lung confirmed by pathological examination. Case 1: 64-year-old male underwent left lower lobectomy with lymph node resection. The patient has been well 27 months after the operation without tumor recurrence. Case 2: was a 75-year-old male, who underwent left upper lobectomy, partial resection of left lower lobe (S6) with lymph node resection. This patient died of aspiration pneumonia 90 days after successful resection of the tumor. In both patients, resected lung tumors were diagnosed to be true carcinosarcoma by histopathological examinations. True carcinosarcoma is defined to contain both cancelous and sarcomatous elements. Sarcomatous elements may differentiate into rhabdomyosarcoma, osteosarcoma and so on, or they may have non-epithelial elements demonstrated by electron microscopy or immunohistochemical studies. We reviewed the 36 cases with carcinosarcoma of the lung reported in Japanese literatures with special consideration of their histopathological findings. The prognosis of the patients with this rare tumor is also discussed according to the TMN stages.

[Evaluation on the perioperative left ventricular function of aortic valve replacement in patients with chronic aortic regurgitation].

To evaluate the perioperative left ventricular function in 36 patients undergoing aortic valve replacement (AVR) in patients with isolated chronic aortic regurgitation (AR), hemodynamic variables before and within the first 24 hours and one month after operation were measured with Swan-Ganz catheter and echocardiography. The patients were divided into two groups based on preoperative left ventricular end-systolic volume index (LVESVI): 5 patients had a LVESVI of 90 ml/m2 and greater (group I), 31 had a LVESVI smaller than 90 ml/m2 (group II). Left ventricular dimension at end-diastole and end-systole, left ventricular end-diastolic volume index at one month after operation significantly decreased in comparison with that of preoperation. However, left ventricular ejection fraction in the two groups were not improved, and left ventricular mass index in group I did not decreased. On the other hand, changes of cardiac index, stroke volume index and left ventricular stroke work index within the first 24 hours after operation were not significantly different between the two groups. The incidence of postoperative premature ventricular contraction in group I was significantly higher than that of group II. However, there was no difficult problems in postoperative management of the two groups. It is suggested that AVR in AR with left ventricular dysfunction can safely be operated and its postoperative management is similar to AVR in AR with no left ventricular dysfunction.

[Experimental study of cadaver heart preservation--examination of the preservation solution and the time limit from death to cardiectomy].

In Japan, we have not been able to perform a heart transplantation for more than 20 years, because a concept of brain death has not been approved ethically or legally. We examined whether cadaver hearts could be used as grafts for heart transplantations, and which of UW solution (UW) and Euro-Collins solution (EC) was more suitable for cadaver heart preservation. Adult Sprague-Dawley rats were divided into 4 groups of 5 each. In Group I, the hearts were harvested under beating. In the other groups, cardiac arrest was derived by exsanguination, and cardiectomy were performed after 15 (Group II, III) or 30 (Group IV) minutes from flattening of arterial blood pressure. Soon after the cardiectomy, UW (10 ml/kg) was infused in Group I, II and IV, and EC (10 ml/min) was infused in Group III. After simple cold storage (0 degree C) for 30 minutes, oxygenated KHB solution (37 degrees C) were perfused in Langendorff mode for 30 minutes, and were switched over to working mode. Aortic pressure and cardiac output were measured for 30 minutes. Following to the measurements, glutaraldehyde was perfused and studied microscopically. All cases could be switched over to working mode in group I and II, but 2 cases in Group III and 4 cases in IV could not make a pressure enough to keep working mode. In hemodynamic and histological study, there were no significant differences between Group I and II, but Group III and Group IV were clearly inferior to Group I and II. In cadaver heart preservation, UW solution was superior to EC solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Long-term results of primary closure for ventricular septal defects in the first year of life.

The long-term results of primary closure for large ventricular septal defects (VSDs) in infants under 1 year of age with severe symptoms were studied over a period of more than 10 years. Between January, 1971 and March, 1982, 49 infants underwent primary closure of a VSD through a right ventriculotomy using complete cardiopulmonary bypass with mild hypothermia. There were four hospital deaths but no late deaths. Two of four infants with residual shunts had a left ventricular-right atrial shunt which necessitated reoperation. Surgical heart block occurred in two infants who recovered sinus rhythm in the late period. The cardiothoracic ratio decreased from 60.5% preoperatively to 50.6% in the late postoperative period. Examination by cardiac catheterization revealed that the pulmonary-to-systemic pressure ratio (Pp/Ps) of 23 patients with a Pp/Ps of over 0.75 fell from 0.89 +/- 0.09 preoperatively to 0.42 +/- 0.12 by 1 month postoperatively, then to 0.27 +/- 0.05 in the late postoperative period. The latest values for the cardiac index and left ventricular ejection fraction were 3.4 l/min per m2 and 64.4%, respectively. More than 10 years after their operation, all the survivors were growing normally and maintaining a good quality of life, which supports our recommendation that primary repair should be performed in the first year of life for infants with large VSDs.

[Analysis of the determinant factors for open heart surgery without blood transfusion by the quantification theory--possibility of application of maximum surgical blood order schedule for open heart surgery].

In order to complete operations without blood transfusion we have chosen means of preoperative autologous blood saving and intraoperative autotransfusion, but we have not always achieved our purpose. We examined 29 patients (13 patients without blood transfusion and 16 with blood transfusion) to analyze the determinant factors as to whether open heart surgery without blood transfusion may be indicated or not, according to the quantification theory (type II) and to examine the possibility to apply the maximum surgical blood order schedule (MSBOS) for the open heart surgery by the quantification theory (type I). The analysis of determinant factors revealed hematocrit (Ht) value before saving of blood (more than 40%) as the best contributor of possibility of non-blood transfusion surgery, followed by the amount of blood loss during operation (less than 600 ml), the amount of saving blood (more than 800 ml), body weight (less than 70 kg), calculated Ht value on the beginning of cardiopulmonary bypass (CPB) (more than 24%), CPB time (less than 120 minutes) and the amount of postoperative blood loss (less than 600 ml). The prospective using blood volume at the operation was precisely calculated by the values of 4 preoperative factors, that is, the amount of saving blood, calculated Ht value on the beginning of CPB, CPB time and body weight. Therefore it is important to increase the amount of preoperative saving blood and decrease the amount of surgical bleeding in order to perform operations without blood transfusion, and is possible to apply the MSBOS for the open heart surgery.