Increased ratio of FoxP3+ regulatory T cells/CD3+ T cells in skin lesions in drug-induced hypersensitivity syndrome/drug rash with eosinophilia and systemic symptoms.

BACKGROUND: Drug-induced hypersensitivity syndrome/drug rash with eosinophilia with systemic symptoms (DIHS/DRESS) is a severe drug eruption accompanied by multiorgan disorders. Several unique aspects of DIHS/DRESS, including herpesvirus reactivation, liver dysfunction and hypogammaglobulinaemia, have similarities to graft-versus-host disease (GVHD). AIM: In this study, we focused on the dynamics of regulatory T cells (Tregs) infiltrating into the skin lesions of DIHS/DRESS and GVHD. METHODS: Skin biopsies were taken from patients with DIHS/DRESS, GVHD, or maculopapular drug eruption. Tregs were detected using immunostaining with anti-FoxP3. RESULTS: The ratio of FoxP3+ T cells to CD3+ T cells was significantly higher in the skin lesions of patients with DIHS/DRESS than in those of patients with GVHD, and was positively correlated with the number of days from disease onset in the acute phase. CONCLUSIONS: The dynamics of Tregs in skin lesions are different between DIHS/DRESS and GVHD, despite there being many similarities between these conditions.

Comparisons of the effects of 12-week administration of miglitol and voglibose on the responses of plasma incretins after a mixed meal in Japanese type 2 diabetic patients.

To compare the effects of miglitol [an alpha-glucosidase inhibitor (AGI) absorbed in the intestine] and voglibose (an AGI not absorbed) on plasma glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide (GIP) levels, 26 and 24 Japanese type 2 diabetic patients were randomly assigned to receive miglitol or voglibose, respectively. After 12-week administration of both drugs, during 2-h meal tolerance test, plasma glucose, serum insulin and total GIP were significantly decreased and active GLP-1 was significantly increased. Miglitol group showed a significantly lower total GIP level than voglibose group. Miglitol, but not voglibose, significantly reduced body weight (BW). In all participants, the relative change in BW was positively correlated with that of insulin significantly and of GIP with a weak tendency, but not of GLP-1. In conclusion, both drugs can enhance postprandial GLP-1 responses and reduce GIP responses. The significant BW reduction by miglitol might be attributable to its strong GIP-reducing efficacy.

Diurnal changes in urinary excretion of IgG, transferrin, and ceruloplasmin depend on diurnal changes in systemic blood pressure in normotensive, normoalbuminuric type 2 diabetic patients.

Previous studies of diabetic patients indicate that increased urinary excretion of certain plasma proteins (molecular radii <55 A), such as IgG, transferrin, and ceruloplasmin, precede the development of microalbuminuria. Moreover, increases in these urinary proteins predict future development of microalbuminuria. To clarify whether blood pressure changes influence urinary excretion of these proteins, we examined relationships between diurnal blood pressure changes measured by ambulatory blood pressure monitoring and urinary excretion of IgG, transferrin, ceruloplasmin, alpha2-macroglobulin (88 A) and albumin (36 A) measured separately during the day and night in 20 healthy controls and 26 normotensive, normoalbuminuric diabetic patients. Diurnal change in systolic blood pressure was not correlated to urinary excretion of either albumin or alpha2-macroglobulin in either diabetic patients or controls. However, statistically significant correlations between diurnal changes in systolic blood pressure and those of urinary excretion of IgG, transferrin and ceruloplasmin were found in diabetic patients but not in controls. The present findings suggest that urinary excretion of IgG, transferrin, and ceruloplasmin are more easily affected than albuminuria by systemic blood pressure changes in normoalbuminuric diabetic patients. This is supported by our previous finding that urinary excretion of IgG, transferrin and ceruloplasmin increased while albuminuria did not following enhanced glomerular filtration rate after acute protein loading, which causes increased glomerular capillary pressure due to afferent arterioles dilation, mimicking diabetic intra-renal hemodynamics. Taken together, these findings suggest that urinary excretion of IgG, transferrin, and ceruloplasmin may be more sensitive indicators of glomerular capillary pressure change than albuminuria in normoalbuminuric diabetic patients.

Continuous decompression using a cannulated ceramic pin for simple bone cysts.

PURPOSE: To report the outcomes of continuous decompression using a cannulated ceramic pin for simple bone cysts (SBCs). METHODS: Seven boys and 3 girls aged 7 to 16 (mean, 11) years with SBCs underwent curettage and continuous decompression using a cannulated ceramic pin. The pin was made of hydroxyapatite and tricalcium phosphate. The clinical course, radiological findings, and complications were retrospectively assessed. RESULTS: The mean follow-up duration was 41 (range, 12-84) months. Five patients were evaluated as 'healed', 2 as 'healing with defect', one as 'persistent cyst', and 2 as 'recurrent cysts'. No peri-operative complications were encountered. One patient had a postoperative fracture at the pin insertion site. Seven patients had pain relief and good outcomes; 2 had a thin cortical rim and complained of occasional pain and their sports activities were restricted. CONCLUSION: Decompression using a cannulated ceramic pin for SBCs is minimally invasive, highly osteoconductive, and does not require bone grafting or a second operation to remove the device.

Attenuated radial augmentation index is associated with successful long-term antihypertensive treatment.

Pulse wave analysis was performed in apparently normal volunteers (n=164) and in essentially hypertensive patients without cardiovascular complications (n=171) using a newly developed non-invasive pulse wave measurement device (HEM-9010AI). Our results suggest that early wave reflections measured by radial augmentation index (AIr) are enhanced in volunteers with systolic blood pressure (SBP) >or= 160 mm Hg compared with the volunteers with their SBP<160 mmHg (98+/-18 vs 88+/-12, P<0.05). Furthermore, AIr is lower in hypertensive patients with long-term antihypertensive treatment than in those with short-term treatment (84+/-10 vs 89+/-13, P<0.01).

Curettage and allograft reconstruction for giant cell tumours.

PURPOSE: To evaluate treatment outcomes in patients with giant cell tumours after curettage and allograft reconstruction and to identify the risk factors for poor oncological and functional outcome. METHODS: 29 patients with giant cell tumours of bone who underwent curettage and allograft reconstruction were retrospectively reviewed. The adjuvants used were heat treatment by electrocautery and hot water. Types of allograft used, time to bone union, complications, functional outcomes, and risk factors for poor function were analysed. RESULTS: The mean time to bone union was 2.8 (range, 1-5) months. In 7 patients the tumours recurred (6 within 2 years); the 5-year recurrence-free survival rate was 77%. Three recurrences were classified as grade III and 4 as grade II; recurrence and the Campanacci grade showed a trend towards association (p=0.06). Tumour in the distal femur was a risk factor for postoperative fracture (p=0.02). Functional outcomes were excellent in 20 patients, good in 6, fair in 2, and a failure in one. The risk factors for poor function were recurrence (p=0.002) and joint instability (p=0.008) but not the Campanacci grade (p=0.10) or postoperative fracture (p=0.76). Lung metastasis, infection, and non-union were not encountered. CONCLUSION: Despite a relatively high recurrence rate (24%), 26 (90%) of the 29 patients had excellent/good functional outcomes. We recommend the use of adjuvants and allografts for the management of giant cell tumours.

Formation of 2'-deoxyoxanosine from 2'-deoxyguanosine and nitrous acid: mechanism and intermediates.

The reaction mechanism for the formation of 2'-deoxy-oxanosine from 2'-deoxyguanosine by nitrous acid was explored using methyl derivatives of guanosine and an isolated intermediate of the reaction. When 1-methylguanosine was incubated with NaNO(2)under acidic conditions, N (5) -methyloxanosine and 1-methylxanthosine were generated, whereas the same treatment of N (2), N (2)-dimethylguanosine generated no product. In a similar experiment without NO(2)(-), participation of a Dimroth rearrangement was ruled out. In the guanosine-HNO(2)reaction system, an intermediate with a half-life of 5.6 min (pH 7.0, 20 degrees C) was isolated and tentatively identified as a diazoate derivative of guanosine. The diazoate intermediate was converted into oxanosine and xanthosine at a molar ratio (oxanosine:xanthosine) of 0.26 at pH 7.0 and 20 degrees C. The ratio was not affected by the incubation pH between 2 and 10, but increased linearly with temperature from 0.22 (0 degrees C) to 0.32 (50 degrees C). The addition of acetone also increased the ratio up to 0.85 (98% acetone). Based on these results, a con-ceivable pathway for the formation of 2'-deoxyoxanosine from 2'-deoxyguanosine by HNO(2)is proposed.

Specific interaction with guanine residues of Z-form DNA by bleomycin-nickel(III) complex.

The Ni(III) complex of bleomycin (BLM) reacts with guanine residues of the oligomer duplex only under high salt condition (4.5 M NaCl) where the (dC-dG)6 sequence exists as the isolated Z-form segment. In addition, the (dC-dG)n inserts were subcloned into a plasmid so that reactivity of the BLM-Ni(III) complex with restriction fragments could be examined. Results are reported for two fragments: an EcoRI-DdeI fragment in which the (dC-dG)n insert is flanked at the 5' end by a segment of B-DNA and a HindIII-DdeI fragment in which the (dC-dG)n fragment is embedded at both ends by stable B-DNA forming segments. The BLM-Ni(III) complex does not bind to (dC-dG)5 sequence of pBRZ10 DNA fragment under any conditions, but to several guanine residues of (dC-dG)8 and (dC-dG)12 sequences of pBRZ16 and pBRZ24 DNA fragments in 4.5 M NaCl solution. In the fragments of pBRZ16 and pBRZ24 DNAs, the BLM-Ni(III) complex reacts with most of guanine bases of (dC-dG)8 and (dC-dG)12.

Selective genotyping with epistasis can be utilized for a major quantitative trait locus mapping in hypertension in rats.

Epistasis used to be considered an obstacle in mapping quantitative trait loci (QTL) despite its significance. Numerous epistases have proved to be involved in quantitative genetics. We established a backcross model that demonstrates a major QTL for hypertension (Ht). Seventy-eight backcrossed rats (BC), derived from spontaneously hypertensive rats (SHR) and normotensive Fischer 344 rats, showed bimodal distribution of systolic blood pressure (BP) values and a phenotypic segregation ratio consistent with 1:1. In this backcross analysis, sarco(endo)plasmic reticulum Ca(2+)-dependent ATPase (Serca) II heterozygotes showed widespread bimodality in frequency distribution of BP values and obviously demonstrated Ht. First, in genome-wide screening, Mapmaker/QTL analysis mapped Ht at a locus between D1Mgh8 and D1Mit4 near Sa in all 78 BC. The peak logarithm of the odds (LOD) score reached 5.3. Second, Serca II heterozygous and homozygous BC were analyzed separately using Mapmaker/QTL. In the 35 Serca II heterozygous BC, the peak LOD score was 3.8 at the same locus whereas it did not reach statistical significance in the 43 Serca II homozygotes. Third, to map Ht efficiently, we selected 18 Serca II heterozygous BC with 9 highest and 9 lowest BP values. In these 18 BC, the peak LOD score reached 8.1. In 17 of the 18, D1Mgh8 genotypes (homo or hetero) qualitatively cosegregated with BP phenotypes (high or low) (P < 0.0001, by chi-square analysis). In conclusion, selective genotyping with epistasis can be utilized for a major QTL mapping near Sa on chromosome 1 in SHR.

Induction therapy by frequent administration of doxorubicin with four other drugs, followed by intensive consolidation and maintenance therapy for adult acute lymphoblastic leukemia: the JALSG-ALL93 study.

In order to improve the disappointing prognosis of adult patients with acute lymphoblastic leukemia (ALL), we applied similar induction therapy as that used for acute myeloid leukemia (AML), ie frequent administration of doxorubicin (DOX). DOX 30 mg/m(2) was administered from days 1 to 3 and from days 8 to 10 together with vincristine, prednisolone, cyclophosphamide and L-asparaginase, followed by three courses of consolidation and four courses of intensification. From December 1993 to February 1997, 285 untreated adult patients with de novo ALL were entered. Of 263 evaluable patients (age 15 to 59; median 31), 205 (78%) obtained complete remission (CR). At a median follow-up period of 63 months, the predicted 6-year overall survival (OS) rate of all patients was 33%, and disease-free survival (DFS) rate of CR patients was 30%, respectively. By multivariate analysis, favorable prognostic factors for the achievement of CR were age <40 and WBC <50 000/microl; for longer OS were age <30 and WBC <30 000/microl; and for longer DFS of CR patients were FAB L1 and ALT <50 IU/l. Among 229 patients who had adequate cytogenetic data, 51 (22%) had Philadelphia (Ph) chromosome. Ph-negative chromosome was a common favorable prognostic factor for CR, longer OS and DFS. DFS was not different between early sequential intensification (n = 48) and intermittent intensification (n = 43) during the maintenance phase. Among CR patients under 40 years old, the 6-year survival was not different between the allocated related allo-BMT group (34 patients) and the allocated chemotherapy group (108 patients). However, among patients with Ph-positive ALL, the survival of patients who actually received allo-BMT was superior to that of patients who received chemotherapy (P = 0.046).

Structural determinants of ion selectivity in brain calcium channel.

Glutamic acid residues in the SS2 segment of the internal repeats III and IV of the brain calcium channel BI were subjected to single point mutations. The mutant channels were tested for macroscopic current properties and sensitivities to inorganic blockers. The mutation that replaces glutamic acid 1,469 with glutamine altered ion-selection properties and strongly reduced the sensitivity to Cd2+, whereas the analogous mutation of glutamic acid 1,765 exerted smaller effects on ion-selection properties. Our results indicate that these glutamic acid residues, equivalently positioned in the aligned sequences, play different roles in the selective permeability of the calcium channel.

A study of CD45RO, CD45RA and CD29 antigen expression on human decidual T cells in an early stage of pregnancy.

The decidua is the place where the fertilized egg is implanted and where the immunocompetent cells of the mother come into direct contact with genetically disparate cells of the conceptus. Although the T cells in the decidua are exposed to fetal antigens, the fetus is not rejected by maternal immunocompetent cells. In the present study, we examined surface markers to determine whether the T cells in the human decidua are naive T cells without or memory T cells with a history of antigen stimulation. Although few T cells were present in the decidua, as compared to the peripheral blood, CD45RO+, CD29+ and CD45RA- CD4+ T cells as well as CD45RO+, CD29+ and CD45RA- CD8+ T cells, which are considered to be memory T cells, were in the majority, with only small numbers of CD45RO-, CD29- and CD45RA+ CD4+ and CD8+ cells, which are naive T cells, present. Also, the decidual mononuclear cells secreted IL-2 and IL-4. Since IL-4 is secreted only by memory T cells, it is suggested that in the decidua memory T cells increase in number and secrete cytokines, thereby in some way influencing the phenomenon of fertility.

Bone formation as a reaction to hydraulic hydroxyapatite thermal decomposition product used as bone cement in rabbits.

The bone activity of hydroxyapatite thermal decomposition product was evaluated by the amount of new bone formation after implantation into the femur of rabbits. New bone formation was observed to be bonded to the entire adjacent surface of implanted ceramics at the metaphysis in 16 out of 20 specimens 12 weeks after implantation.

Human anti-murine antibodies interfere with CPR assays performed with commercial kits.

Quantitation of C-peptide is important for the assessment of insulin secretion, in particular in patients receiving insulin therapy. Since the CPR levels become much higher than the concentration of C-peptide for several reasons, such as the high concentration of proinsulin, CPR values sometimes need to be assessed carefully. We have had two diabetic patients whose CPR values were abnormally high when determined with a Daiichi C-peptide kit III (method 1). CPR values determined by other methods were from two to ten times lower, indicating considerable interference when method 1 was used. Since method 1 uses mouse monoclonal antibodies (mmab) for detection antibodies, we suspected that human anti-murine antibodies (HAMA) were responsible for the interference. HAMA were detected in serum from both patients (45 and 460 ng/ml in case 1 and case 2 (at peak), respectively). Removal of HAMA from serum eliminated the interference. Modification of method 1 to exclude mmab from the assay system removed all interference. HAMA were, therefore, considered to be the cause of the interference. In case 2, the peak concentration of HAMA was recorded 16 months earlier than the maximum of interference. Further analysis revealed that HAMA with high affinities were responsible for the interference.

Influence of ring opening-closure equilibrium of oxanine, a novel damaged nucleobase, on migration behavior in capillary electrophoresis.

Oxanine (Oxa) is a novel damaged nucleobase which is generated from guanine by HNO2 or NO. As a fundamental study for detection of Oxa formed in vivo, capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) have been tested by changing the pH of the running buffer. At pH 7, CZE did not separate Oxa from seven other nucleobases, and MEKC separated Oxa but their peaks migrated close together. In both the techniques, an extreme peak broadening occurred for Oxa around pH 9 and a good peak separation was achieved at pH 12. The behavior of the Oxa peak is discussed in relation to the unique multistep acid-base equilibria of Oxa.

In vitro chemosensitivity of human soft tissue sarcoma.

Chemotherapy is essential in the treatment of small round cell sarcomas. However, as yet there is no progress concerning the efficacy of chemotherapy in the treatment of other types of soft tissue tumors (STS). The Histoculture Drug Response Assay (HDRA) is an in vitro chemosensitivity test that has a high correlation with clinical response, the usefulness of which has been reported in various kinds of solid tumors. However, there has never been a report on its use in STS until now. In this study, in order to investigate the variation in chemosensitivity in STS, fresh biopsy or surgical samples of STS were tested using the HDRA method. Drug sensitivity testing by HDRA showed that two drugs, ADM and THP, had a significantly higher inhibitory rate than CDDP, IFOS, or VP-16 in the thirty-three soft tissue sarcomas tested. Depending on the morphological type, spindle cell sarcomas were sensitive to THP, which showed significantly higher inhibition rates than CDDP, IFOS, or VP-16. Small round cell sarcomas were relatively sensitive to all of the drugs tested. However the drug sensitivity of pleomorphic cell sarcoma was low except for ADM and THP, while its sensitivity to THP was higher than about 70%. However, there are numerous other soft tissue sarcomas that do not belong to these categories; drug sensitivity testing in each of them and the devising of individualized treatment strategies seems necessary to improve the therapeutic outcome.

Prognostic relevance of urokinase type plasminogen activator, its receptor and inhibitors in chondrosarcoma.

The plasminogen activation system plays an important role in enhancing pericellular proteolysis of tumor invasion/metastasis and in autocrine/paracrine tumor growth stimulation. To investigate the prognostic significance of the plasminogen activation system in human chondrosarcoma, the immunohistochemical expression of urokinase-type plasminogen activator (uPA), urokinase-type plasminogen activator receptor (uPAR), plasminogen activator inhibitor, 1 (PAI-1) and 2 (PAI-2) were analyzed in 28 patients with chondrosarcoma. In multivariate survival analysis, histological grade (p = 0.0008) and location (p = 0.02) were independent risk factors for local relapse. For metastasis-free survival, uPA index (p = 0.006) and PAI-2 index (p = 0.04) were independent prognostic factors. PAI-2 index (p = 0.02), uPAR index (p = 0.02) and histological grade (p = 0.03) predicted total survival. These results demonstrated the usefulness of uPA, uPAR and PAI-2 expression as biological prognostic indicator and the importance of the plasminogen activation system in tumor progression and metastasis in chondrosarcoma.

High tibial osteotomy with fixation by a blade plate for medial compartment osteoarthritis of the knee.

Surgical techniques were described on tibial osteotomy above the tuberosity, the fragments of which were fixed with a Koshino blade plate. Postoperative clinical results on 176 knees with medial compartment osteoarthritis of 138 patients with an average age of 62.1 years were excellent in 110 and good in 55 knees with an average of 5.5 years' follow-up. The best knee score was obtained in the knee with postoperative limb alignment of 6 to 15 degree valgus angulation (standing). The results were satisfactory, especially in postoperative management, which was much easier with the Koshino blade plate fixation than with other methods. The sinking of the osteotomy site was checked through observation of screw movement in the slot of the plate during postoperative management in 53 selected knees, and the averaged distance of sinking was 3.9 +/- 2.5 mm. The time of mechanical bone union judged by cessation of sinking averaged 12.9 +/- 6.2 weeks after osteotomy.