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Background: There have been few reports of clinical pathway (CP) for treatment of asthma attack, because patients with asthma always admit emergently and the severity varies. We introduced CP so that standard asthma treatment can be widely used, and investigated its clinical usefulness., Methods: We designed a new CP for treating asthma attack according to the guideline (Japanese guideline (JGL) and Global Initiative for Asthma (GINA)). 136 patients who admitted to our hospital due to asthma attack from January 1999 to November 2006, were enrolled our study. Excluding cases complicated with pneumonia, COPD or cardiac failure, we evaluated 46 cases treated with the CP comparing with 19 cases treated without the CP. The clinical evaluations include systemic and inhaled steroid use, FEV1.0%, history of asthma, and the duration of asthma attack. Furthermore, we investigated difference between cases with and without prolonged admission., Results: While the rates of systemic and inhaled steroid use in cases without the CP were 57.9% and 52.6% respectively, those in cases with the CP were approximately 100%. Employing the CP, FEV 1.0% at discharge time was elevated from 71.7% to 76.3% and the duration of hospitalization was shortened from 14.2 days to 11.5 days. Mean age of the cases with prolonged admission was higher than the rest., Conclusion: The asthma CP is an effective way for the standard treatment according to the guideline to be used widely even by doctors who are not familiar with asthma treatment. It improves the efficacy of in-hospital treatment.

Infection of the monocyte/macrophage lineage with feline infectious peritonitis virus (FIPV) is enhanced in the presence of anti-FIPV antibodies (antibody-dependent enhancement or ADE). We investigated the following unclear points concerning ADE of FIPV infection: (i) involvement of the virus receptor, feline aminopeptidase N (fAPN), in ADE activity in FIPV infection; (ii) necessity of acidification of the endosome in cellular invasion of FIPV. Virus receptor-blocking experiments using anti-fAPN antibodies at 4 or 37 degrees C and experiments using fAPN-negative U937 cells revealed that fAPN is not involved in ADE of FIPV infection. Experiments using lysosomotropic agents clarified that acidification of the endosome is necessary for cellular invasion by FIPV, regardless of the presence or absence of antibodies. These findings may be very important for understanding the mechanism of ADE of FIPV infection.

We report a case of Wegener's granulomatosis (WG) with central nervous system (CNS) involvement in a woman who complained of bilateral visual disturbance. The intracranial necrotizing granulomatous lesion was confirmed by MR imaging and brain biopsy. After high-dose oral corticosteroid treatment, not only clinical manifestations but also laboratory tests improved. Moreover, the CNS lesion completely regressed. This suggests that high-dose corticosteroid alone might be effective in treating WG with CNS involvement.

We evaluated the diagnostic value of anti-cyclic citrullinated peptide 2 (anti-CCP2) antibodies and other potential diagnostic biomarkers (IgM rheumatoid factor, anti-agalactosyl IgG antibodies, matrix metalloproteinase 3, C-reactive protein) for predicting early development of rheumatoid arthritis (RA). Patients were defined as having recent-onset undifferentiated arthritis (UA) if they had developed arthritis in two or more joints within the previous 2 years and could not be classified with a well-defined arthropathy. Baseline levels of biomarkers were measured in blood samples collected at the entry of the study and the patients were followed for 1 year to monitor development of RA. Diagnoses of RA and non-RA arthropathies were made according to individual standard diagnostic criteria. A total of 146 patients were enrolled in the study. In the follow-up year, 18 patients developed RA, 54 developed non-RA arthropathies, and 60 remained in the UA category. The sensitivity and specificity of the presence of anti-CCP2 antibodies for the diagnosis of RA were 83.3 and 93.0%, respectively. The positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of anti-CCP2 antibodies for RA (65.2, 97.2, and 91.7%, respectively) were higher than for any other biomarker. Combination of anti-CCP2 with any other biomarker only slightly improved each diagnostic value compared to the presence of anti-CCP2 alone. Among the anti-CCP2-positive patients, the average titer was significantly higher in those with RA than in non-RA or UA patients (163.7 +/- 138.4 vs 55.2 +/- 72.0 U/ml, p = 0.017). Anti-CCP2 antibodies are superior to any other single biomarker for predicting early development of RA in patients with recent-onset UA and the diagnostic value of anti-CCP2 alone is similar to that for biomarker combinations. Moreover, the anti-CCP2 antibody titer is useful to discriminate between patients at high risk for early developing RA from those at risk of developing non-RA arthropathies.

Objective: Recently, small semiconductor gamma cameras (SSGCs) with high resolution and sensitivity, which are much more convenient to use as compared with SPECT and PET, have been developed for mapping the sentinel lymph node. The high resolution and sensitivity of the SSGCs may make them useful for small animal imaging. Therefore, we assessed the applicability of the SSGC to small animal imaging using a rat model of focal cerebral ischaemia., Methods: The right middle cerebral artery (MCA) of anaesthetized rats was occluded intraluminally with a nylon monofilament. Twenty-four hours after the occlusion, 99mTc-HMPAO (3.7 MBq) was injected and a static acquisition (5 min) was performed using the SSGC. Regions of interest (ROIs) were set on each hemisphere of the horizontal brain images. After the acquisition, the brains were removed and the radioactivity in each hemisphere was measured using an NaI scintillation counter., Results: Reduced CBF in the right MCA territory was clearly visualized with the SSGC in vivo. The radioactivity in the ROIs determined by the SSGC was significantly correlated with that determined by the ex vivo counting method (P<0.001, R2=0.74). Furthermore, in both of the in-vivo imaging and ex-vivo counting methods, the right to left count ratio (R/L ratio) was significantly lower in the MCA-occluded rats than that in normal rats (MCA-occluded rats: 0.77+/-0.08, normal rats: 1.01+/-0.07, P<0.005)., Conclusions: The SSGC clearly visualized and quantitatively detected the reduced CBF in MCA-occluded rats. Furthermore, these high resolution and sensitivity of SSGC can avoid the disadvantage of small animal imaging with PET and SPECT, such as a large mass injected tracer and the exposure of investigators to radiation. Thus, the high resolution and sensitivity of the SSGC make it useful for small animal imaging.

The threshold of cerebral blood flow (CBF) into infarction in rats has been indicated to be similar to that in patients. However, CBF does not reflect metabolic function, and so estimations of oxygen metabolism have been required. Here, we estimated changes in oxygen metabolism after occluding the right middle cerebral artery (MCA) in rats using an injectable (15)O-O(2) we developed. A decrease in CBF (left: 0.67+/-0.22 mL/min/g, right: 0.44+/-0.17 mL/min/g, P<0.05) and compensatory increase in the oxygen extraction fraction (OEF) (left: 0.42+/-0.13, right: 0.50+/-0.19, P<0.05) were observed at 1-h after occlusion. In contrast, a marked decrease in CBF and the cerebral metabolic rate for oxygen and a collapse of the compensatory OEF mechanism were found at 24 h after occlusion. Injectable (15)O-O(2) could be used to reliably estimate oxygen metabolism in an infarction rat model with positron emission tomography.

Unlabelled: We showed that CD9, a member of tetraspanin superfamily proteins, is expressed in a specific membrane microdomain, called "lipid raft," and is crucial for cell fusion during osteoclastogenesis after activation of the RANK/RANKL system., Introduction: Osteoclasts are bone-resorbing multinuclear polykaryons that are essential for bone remodeling and are formed through cell fusion of mononuclear macrophage/monocyte lineage precursors. Although osteoclastogenesis has been shown to be critically regulated by the RANK/RANKL system, the mechanism how precursor cells fuse with each other remains unclear. We examined the function of CD9, a member of tetraspanin superfamily, which has previously been shown to form macromolecular membrane microdomains and to regulate cell-cell fusion in various cell types., Materials and Methods: We used RAW264.7, a macrophage/monocyte lineage cell line, which can differentiate into osteoclast-like polykaryons on the application of RANKL. Expression and distribution of CD9 was assessed by Western blotting, fluorescence-assorted cell sorting (FACS) and immunohistochemistry with light and electron microscopy. A specific neutralizing antibody and RNA interference were used to inhibit the function of CD9, and green fluorescent protein (GFP)-CD9 was exogenously expressed to enhance the effect of CD9. The distribution of CD9 in lipid microdomain was examined by biochemical (sucrose density gradient) isolation and imaging technique., Results: CD9 is expressed on cell surfaces of RAW264.7, which is enhanced by RANKL. Targeted inhibition of CD9 decreases the number of osteoclast-like cells. On the other hand, overexpression of CD9 promotes spontaneous cell fusion even in the absence of RANKL. CD9 is localized in detergent-insoluble "lipid raft" microdomain in RANKL stimulation, and disruption of lipid rafts markedly reduces the formation of osteoclast-like polykaryons. Immunohistochemical studies of bone tissues revealed the expression of CD9 in osteoclasts in vivo., Conclusions: These data suggest that function of tetraspanin CD9 and its expression in lipid rafts are crucial for cell fusion during osteoclastogenesis.

A 15-year-old Japanese girl developed bulbar palsy and upper limb-dominant muscle weakness 2 weeks after the onset of an upper respiratory tract infection due to cytomegalovirus (CMV). Her symptoms resembled that seen in the pharyngeal-cervical-brachial variant (PCB) of Guillain-Barré syndrome (GBS). Although bulbar palsy usually continues for several months in PCB, her bulbar palsy was very mild and improved rapidly before intravenous immunoglobulin therapy was instituted. Serum anti-GT1a IgG antibody titer was elevated at the acute phase of the disease and gradually decreased. The bulbar palsy-dominant GBS is thought to relate to anti-GT1a antibody and Campylobacter jejuni infection in adult patients. Our Case report suggests that CMV can also induce the production of anti-GT1a antibody, thereby resulting in PCB. When one sees acute onset bulbar palsy and limb muscle weakness, the possibility of PCB, even in children, should be considered, thus compelling the need for serum anti-ganglioside antibody measurement.

Immune complex and complement systems play an important role in membranoproliferative glomerulonephritis (MPGN). X-linked agammaglobulinemia (XLA) is a primary immunodeficiency characterized by severe hypogammaglobulinemia. We report the case of an XLA patient who developed MPGN during an intravenous immunoglobulin (IVIG) treatment. In this patient, the serum IgG level was maintained at more than 400 mg/dl of regular IVIG administration (2.5 g/dose/month). The patient presented with microscopic hematuria, proteinuria (U-pro/Cr: 4.0-4.2) and low serum complement levels (C3: 57.8 mg/dl) 3 years after IVIG treatment and was diagnosed histopathologically as having MPGN type III. Both hematuria and proteinuria significantly improved, and the serum complement level returned to a normal level following methylprednisolone pulse therapy. To our knowledge, this is the first case report of MPGN associated with XLA. Although it is unclear how MPGN occurred in this XLA patient, we suggest that residual humoral immunity in the patient could be associated with the development of MPGN.

Unlabelled: Although several COX-2 inhibitors have recently been radiolabeled, their potential for imaging COX-2 expression remains unclear. In particular, the sulfonamide moiety of COX-2 inhibitors may cause slow blood clearance of the radiotracer, due to its affinity for carbonic anhydrase (CA) in erythrocytes. Thus, we designed a methyl sulfone-type analogue, 5-(4-iodophenyl)-1-[4-(methylsulfonyl)phenyl]-3-trifluoromethyl-1H-pyrazole (IMTP). In this study, the potential of radioiodinated IMTP was assessed in comparison with a (125)I-labeled celecoxib analogue with a sulfonamide moiety ((125)I-IATP)., Methods: The COX inhibitory potency was assessed by measuring COX-catalyzed oxidation by hydrogen peroxide. The biodistribution of (125)I-IMTP and (125)I-IATP was determined by the ex vivo tissue counting method in rats. Distribution of the labeled compounds to rat blood cells was measured., Results: The COX-2 inhibitory potency of IMTP (IC(50) = 5.16 microM) and IATP (IC(50) = 8.20 microM) was higher than that of meloxicam (IC(50) = 29.0 microM) and comparable to that of SC-58125 (IC(50) = 1.36 microM). The IC(50) ratios (COX-1/COX-2) indicated the high isoform selectivity of IMTP and IATP for COX-2. Significant levels of (125)I-IMTP and (125)I-IATP were observed in the kidneys and the brain (organs known to express COX-2). The blood clearance of (125)I-IMTP was much faster than that of (125)I-IATP. Distribution of (125)I-IATP to blood cells (88.0%) was markedly higher than that of (125)I-IMTP (18.1%), which was decreased by CA inhibitors., Conclusions: Our results showed a high inhibitory potency and selectivity of IMTP for COX-2. The substitution of a sulfonamide moiety to a methyl sulfone moiety effectively improved the blood clearance of the compound, indicating the loss of the cross reactivity with CA in (125)I-IMTP. (123)I-IMTP may be a potential SPECT radiopharmaceutical for COX-2 expression.

A 47-year-old man who had been taking minocycline for palmoplantar pustulosis developed fever, myalgias, polyneuropathy, and testicular pain, with elevated C-reactive protein (CRP). Neither myeloperoxidase- nor proteinase-3-antineutrophil cytoplasmic antibody was positive. These manifestations met the American College of Rheumatology 1990 criteria for the classification of polyarteritis nodosa. Stopping minocycline led to amelioration of symptoms and normalization of CRP level. To our knowledge, this is the second case of minocycline-induced vasculitis satisfying the criteria. Differential diagnosis for drug-induced disease is invaluable even for patients with classical polyarteritis nodosa.

A 7-year-old boy, who was noted to be a slow runner at the age of 2 years, had progressive muscle weakness and atrophy, preferentially affecting distal muscles. At 3 years of age, he had scoliosis and difficulty in standing on tip-toe. Serum creatine kinase was 1074IU/l. Muscle CT scan showed low-density areas in the lower legs and upper arms, but predominantly in the gastrocnemius and soleus muscles. Biopsy of the biceps brachii muscle showed moderate dystrophic changes with normal dysferlin expression on immunohistochemical and western blot analyses. Although muscle involvement mimicked that seen in Miyoshi myopathy (MM), the very early onset of the disease and scoliosis were quite unusual for MM. We, therefore, made the diagnosis of early onset dysferlin-positive distal muscular dystrophy, probably a new type of distal muscular dystrophy.

A 62-year-old man admitted to our hospital because of dry eyes and thirst. On admission, his bilateral parotid and submandibular glands were swelling. Laboratory data revealed severe eosinophilia (3309/microl), elevated serum IgE (1821IU/dl) and hypergammaglobulinemia (IgG 6049 mg/dl). Interstitial pneumonia and renal dysfunction were present. He was diagnosed with Sjögren's syndrome (SjS) according to the European criteria. After the initiation of the therapy with 30 mg of prednisolone, all abnormal findings including salivation ameliorated promptly. In the absence of any other obvious causes of eosinophilia and elevation of serum IgE, these findings might be associated with SjS itself. Moreover, both serum IL-5 and IgG4 levels were elevated in this patient, suggesting that Th2 type cytokines were dominant. Elevation of serum IgE and IgG4, and hypereosinophilia were likely to be reactive. To our knowledge, there have been no reports of these cytokines and of IgG subclass in such patients. We propose the existence of Th2-type-cytokine-dominant subtype of SjS.

We have identified the genes whose expressions are augmented in the blood cells of the patients with systemic lupus erythematosus (SLE) using the 'stepwise subtraction' technique along with microarray analysis. The expression levels of these genes were assessed by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) in 31 SLE patients and 30 healthy controls. We found that the transcription levels of following eight genes were significantly increased in SLE patients; interferon (IFN)-alpha-inducible protein 27 (IFI27), IFN-alpha-inducible protein IFI-15K (G1P2), IFN stimulated gene 20 kDa (ISG20), epithelial stromal interaction 1 (EPSTI1), defensin-alpha (DEFA3), amphiregulin (AREG) and two genes of unknown function (BLAST accession nos AL050290 and AY358224 = SLED1). In comparison with idiopathic thrombocytopenic purpura (ITP), an organ-specific autoimmune disease, IFI27, G1P2 and SLED1 were preferentially upregulated in SLE. In contrast, AREG and AL050290 were more highly expressed in ITP than in SLE. We correlated changes in gene expression and clinical/laboratory features of SLE and found that expression of ISG20, EPSTI1 and SLED1 are significantly correlated with lymphocyte counts. Genes linked to IFN are well known to influence SLE, but several other novel genes unrelated to IFN signaling we report here would be useful to understand the pathophysiology of SLE.

We investigated the inductive activity of infective influenza A/PR/8/34 (PR8) virus and its ether-split product (ESP) on the expression of inducible nitric oxide (NO) synthase (iNOS) and NO production in RAW264.7 (RAW) cells, a murine macrophage (M psi) cell line, and thioglycolate-elicited peritoneal M psi (TPM). In both cells, PR8 virus infection induced iNOS mRNA between 4 hr and 24 hr, attaining a peak value at 12 hr. In correlation with induction of iNOS mRNA, NO amounts increased significantly from 12 to 24 hr. Moreover, this study demonstrated that ESP with the same hemagglutination titer as PR8 virus could induce iNOS mRNA and NO production, although the inductive activity of ESP was weaker than that of PR8 virus. Considering the dual role (beneficial and detrimental roles) of NO on certain inflammatory disorders and virus infections, the inductive activity of influenza virus on the iNOS-mediated NO production independent of its infectivity might contribute to a modification of influenza virus infection.

For patients with pollinosis, self-care is important as well as medical care. In this study, we investigated the real situation of patients with pollinosis-how they manage their symptoms. A questionnaire-based examination was conducted among 488 patients who visited National Osaka-Minami hospital and Sano ENT clinic. The onset of pollen dispersion occurred 17th Feb. 34.6% of patients visited before the onset of pollen dispersion. 45.5% of them had a checkup for allergens. 84.8% of them utilized pollen information. 41.4% of them did self-care. The rate of patients who utilized pollen information and did self-care was higher in female compared with male. The rate of those who did self-care was significantly higher in the patients who visited before pollen dispersion, who had a checkup for allergens, who utilized pollen information, compared with the patients did not. It is necessary to show the information of self-care as well as pollen, and suggest the importance of self-care against pollen allergens.

Adult Still's disease (ASD) is a chronic multisystemic disease. Extraordinarily high serum levels of IL-18 in ASD patients have been described, whereas the mechanism remains to be clarified. This study aimed to evaluate proinflammatory cytokines and to consider their pathological roles. In patients with rheumatic diseases (n = 151), blood samples were taken at the active phase and the serum levels of IL-18 and other proinflammatory cytokines were measured by ELISA. The extra-high levels of IL-18 were confirmed selectively in ASD patients (n = 10). In the active phase of ASD patients, the levels of IL-6 were elevated accordingly, but IL-1beta and TNF-alpha were undetectable. As to Th1-Th2 cytokines, the levels of IL-4 and IL-13, but not INF-gamma, IL-12, or IL-2, were elevated in all ASD patients examined. Moreover, the serum levels of IL-18 showed a good correlation with those of IL-4, suggesting that ASD reflects a Th2 rather than a Th1 cytokine profile.

Unlabelled: A standard treatment with a dose of 2 g/kg of intravenous immune globulin (i.v.IG) is sometimes ineffective in some patients with Kawasaki disease (KD), who are commonly treated with additional i.v.IG. An interval of 6 to 7 months in Japan and 11 months in the United States is recommended before vaccination against measles after i.v.IG treatment for KD, but it is not known how long after treatment with additional i.v.IG. We studied the persistence of measles antibody titers in six episodes of KD in five patients without history of measles infection or vaccination, 4 to 28 months of age, after additional infusion of i.v.IG. The total dose was 4 g/kg in five episodes and 6 g/kg in one episode. Enzyme immunoassay antibody titers against measles were still positive (> or =400) in all patients tested 3 months after additional infusion of i.v.IG and positive in one patient and equivocal (> or =200 and <400) in three patients after 6 months, but negative (<200) in all after 9 months following infusion. Neutralization test antibody titers against measles were still positive (> or =1:4) in all patients 3 months after additional infusion of i.v.IG, but only one after 6 months, and negative (<1:4) in all after 9 months following infusion., Conclusion: We suggest that an appropriate interval between infusion of 4 g/kg of intravenous immune globulin and measles vaccination be 9 months. The 11-month interval recommended in the United States for 2 g/kg may be longer than necessary.

We have investigated the effect of Zingiber offifinale Rosc. (ZOR) on macrophage-inducible nitric oxide (NO) synthase (macNOS) mRNA expression and NO production in RAW264.7 cells, a murine macrophage cell line; 100 microg/ml ZOR can induce macNOS mRNA expression, but induction effects at a dose below 10 microg/ml were weak or negligible. Kinetic studies showed that macNOS mRNA can be detected from 4 hours to 24 hours after dosing, with a peak at 8 hours. In accordance with the induction of macNOS mRNA expression, NO concentrations increased from 3.4 microM at 2 hours to almost 150 microM at 24 hours, reflecting a longer period of macNOS mRNA expression. The activity of ZOR can be considered to contribute, at least in part, to the beneficial effects of ZOR through the macNOS-mediated activation of the biodefense mechanism.

In this study, we examined osteopontin (OPN) production in myeloma cells and plasma OPN levels in multiple myeloma (MM) patients. We assessed OPN production in bone marrow cells (BMCs) by immunocytochemistry and enzyme-linked immunosorbent assay (ELISA). We also assessed OPN production in various B-cell malignant cell lines, including three myeloma cell lines by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. In addition, we measured plasma OPN concentrations by ELISA in 30 MM patients, 21 monoclonal gammopathy of undetermined significance (MGUS) patients and 30 healthy volunteers. As a result, in an immunocytochemical study, abundant OPN was detected in BMCs from overt MM patients, whereas no OPN was detected in BMCs from patients with other haematological diseases, including MGUS. Cultured BMCs from overt MM patients produced more OPN than those from patients with either smouldering MM or MGUS. Myeloma cell lines spontaneously produced OPN. Plasma OPN levels of MM patients were significantly higher than those of MGUS patients and healthy volunteers (P < 0.05). Moreover, they correlated with both progression and bone destruction of the disease (P < 0.05). These suggest that myeloma cells actively produce OPN, which possibly contributes to osteoclastic bone resorption in MM. Plasma OPN levels may be a useful biomarker for assessing bone destruction in MM and distinguishing MM from MGUS or smouldering MM.

Here we present a rare case of systemic sclerosis (SSc) with idiopathic portal hypertension (IPH) having factor V Leiden mutation, a well-known genetic risk factor for various venous thromboses. A 53-year-old SSc patient showing huge esophageal varices and splenomegaly without liver cirrhosis was diagnosed with IPH. As heterozygous Leiden mutation was detected, some coagulation abnormality and resultant formation of microthrombi in the branches of portal vein were suggested as a cause of IPH in this case. This is the first report showing the possible association between Leiden mutation and one of the complications of SSc.

Objective: To investigate the involvement of osteopontin (OPN) in the pathogenesis of rheumatoid arthritis (RA), localization and production of OPN were examined in patients with RA., Methods: Localization of OPN in the rheumatoid synovium was examined by immunohistochemistry. In vitro OPN production by cultured synovial cells from patients with RA (n = 5) and with osteoarthritis (OA) (n = 5) was assessed by ELISA. OPN concentrations in plasma and synovium were quantified in patients with RA (n = 23) by 2 distinct ELISA systems to measure both thrombin cleaved and non-cleaved OPN. The same experiments were done in patients with OA (n = 15) and healthy volunteers (n = 10) as a control., Results: OPN was highly detected by immunohistochemistry predominantly in the RA synovial lining cells, while less and scattered OPN was detected in OA synovial tissues. ELISA revealed that cultured RA synovial cells secreted significantly more OPN than OA cells. ELISA also showed a marked increase of OPN levels in synovial fluid (SF) of patients with RA and with OA compared to the control plasma OPN levels, although OPN levels were not increased in RA and OA plasma compared to healthy controls. SF OPN levels of patients with RA were significantly higher than those of patients with OA, and correlated with serum C-reactive protein levels. The ratios of thrombin cleaved versus non-cleaved OPN were significantly increased in RA plasma and SF compared with OA plasma and SF and plasma from healthy controls., Conclusion: Our results revealed enhanced local production of OPN in rheumatoid joints, suggesting involvement of OPN in the pathogenesis of RA.

Infectious mononucleosis (IM) is an acute sporadic infection that usually affects young adults, and during infection a massive expansion of CD8 T cells is generally considered to occur. However, CD28 expression of the expanded cells has not been characterized. When peripheral blood mononuclear cells of acute IM (AIM) patients were analyzed by flow cytometry, a continuous spectrum of CD28 intensity ranging from negative to high, which could be separated into CD28 negative, intermediate (int), and positive, was seen for CD8 T cells. We studied 26 IM patients who were diagnosed on the basis of standard methods and found that all patients had the continuous CD28 spectrum. CD28 is a costimulatory molecule on T cells, and its expression is associated with the subdivision of CD8 cells into cytotoxic (CD28-positive) and suppressor (CD28-negative) T cells. After 24 h of ex vivo culturing, however, the continuous spectrum was found to consist of only CD28-positive and CD28-negative CD8 T cells, because the CD28-int cells had disappeared due to apoptosis. The CD28-int T cells have several cytotoxic functions, suggesting that CD28-int T cells are effectors. Examination of other costimulatory markers in AIM patients showed that CD80 and CD152 were not affected. In patients with other viral infections, such as measles or rubella, however, the continuous spectrum was not detected. These results suggest that there is an unusual CD28 expression pattern in patients with AIM, namely, the presence of a functional CD28-int subset among CD8 T cells. These findings are of special importance for clarifying the defense mechanism against Epstein-Barr virus infection, and the role of CD28 molecules in humans and should also be helpful for the diagnosis of AIM.

It has been previously reported that green-tea extract (GTE) inhibits the growth of influenza virus by preventing its adsorption. In this study, we further investigated whether GTE exerts an additional inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS) and thereby inhibits the growth of influenza A and B viruses in Madin-Darby canine kidney cells. The vital fluorescence microscopic study showed that GTE inhibited acidification of ELS in a concentration-dependent manner. Moreover, the growth of influenza A and B viruses was equally inhibited when the cells were treated with GTE within as early as 5 to 15 min after infection, depending on the virus strains. The fact that (-)epigallocatechin (EGC), one of major catechin molecules in GTE, exerts the inhibitory effects on the acidification of ELS and virus growth in a manner similar to that of GTE strongly suggests that EGC is one of the active components in the extract.

Functional roles of interleukin (IL-)6 in T cell response were investigated. Mice deficient in IL-6 and wild mice were immunized with antigens (myelin oligodendrocyte glycoprotein or methylated BSA) and production of IL-4 and interferon (IFN)-gamma by regional lymph nodes was measured. IL-6 deficiency led to an enhancement of IL-4 and an inhibition of IFN-gamma production. Moreover, polyclonal stimulation of spleen T cells from unimmunized IL-6-deficient mice with anti-CD3 plus anti-CD28 antibodies (Abs) demonstrated an enhancement of T helper (Th)(2)responses. The presence of IL-6, however, augmented IL-4 production but it inhibited IFN-gamma expression by spleen T cells in response to polyclonal stimulation and by antigen-primed spleen T cells in response to re-challenge with the antigen. In contrast, the induction of spleen CD4-positive T cells into Th(2)cells in vitro by the anti-CD3 plus IL-4 was completely suppressed by exogenously added IL-6, whereas Th(1)differentiation of T cells by the anti-CD3 plus IL-12 was not inhibited by the presence of IL-6. Thus, these results indicate that IL-6 physiologically could modulate qualitative T cell response and suggest that it augments Th(1)responses partly through its inhibitory capability of IL-4-induced Th(2)differentiation of naive T cells., (Copyright 2001 Academic Press.)

Objective: To investigate the roles of interleukin-6 (IL-6) in the pathogenesis of rheumatoid arthritis (RA) by studying its effect on murine collagen-induced arthritis (CIA)., Methods: IL-6-deficient (IL-6-/-) mice with a genetic background of susceptibility to CIA were generated by backcrossing them with DBA/1J mice for 8 generations. Clinical and immunologic features were compared between these mice and IL-6 wild-type (IL-6+/+) littermates with CIA., Results: Serum IL-6 levels increased during the development of CIA in IL-6+/+ mice. Two prominent peaks were observed. The first was coincident with the onset of arthritis, and the second one was observed during exacerbation of the disease. The onset of arthritis in IL-6-/- mice was delayed for 2 weeks compared with that in IL-6+/+ mice, and the severity of arthritis, as indicated by the arthritis score, remained significantly lower in IL-6-/- mice during the entire followup period (14 weeks), although all IL-6-/- mice developed definite arthritis as did the IL-6+/+ mice. Histologic severity was also reduced in IL-6-/- mice. In addition, radiologic changes such as osteopenia and bone erosion were reduced significantly in these animals. Both humoral and cellular responses to type II collagen (CII) in IL-6-/- mice were reduced to about half those in IL-6+/+ mice. In addition, enhanced production of IL-4 and IL-10 in response to concanavalin A stimulation was observed in IL-6-/- mice., Conclusion: IL-6 plays an important role in the development of CIA, and both suppression of specific immune responses to CII and a tendency to a shift toward a Th2 cytokine profile might contribute in part to the attenuation of CIA in IL-6-/- mice. These findings suggest that blockade of IL-6 might be beneficial in the treatment of RA.

To investigate the direct role of interleukin (IL) 6 in the development of rheumatoid arthritis, IL-6-deficient (IL-6 -/-) mice were backcrossed for eight generations into C57BL/6 mice, a strain of mice with a genetic background of susceptibility for antigen-induced arthritis (AIA). Both histological and immunological comparisons were made between IL-6-deficient (IL-6 -/-) mice and wild-type (IL-6 +/+) littermates after the induction of AIA. Although all IL-6 +/+ mice developed severe arthritis, only mild arthritis was observed in IL-6 -/- mice. Safranin O staining demonstrated that articular cartilage was well preserved in IL-6 -/- mice, whereas it was destroyed completely in IL-6 +/+ mice. In addition, comparable mRNA expression for both IL-1beta and tumor necrosis factor alpha, but not for IL-6, was detected in the inflamed joints of IL-6 -/- mice, suggesting that IL-6 may play a more crucial role in cartilage destruction than either IL-1beta or tumor necrosis factor alpha. In immunological comparisons, both antigen-specific in vitro proliferative response in lymph node cells and in vivo antibody production were elicited in IL-6 -/- mice, but they were reduced to less than half of that found in IL-6 +/+ mice. Lymph node cells of IL-6 -/- mice produced many more Th2 cytokines than did IL-6 +/+ mice with either antigen-specific or nonspecific stimulation in in vitro culture. Taken together, these results indicate that IL-6 may play a key role in the development of AIA at the inductive as well as the effector phase, and the blockade of IL-6 is possibly beneficial in the treatment of rheumatoid arthritis.

A 60-year-old woman who had undergone surgery of oral cancer under general anesthesia developed an attack of acute angle-closure glaucoma the next morning. Her intraocular pressure decreased immediately by adequate treatments including surgical procedures (laser iridotomy and trabeculectomy), and her symptom improved. It is considered that this event was induced by several factors such as atropine given before and during general anesthesia, perioperative hypertension and anatomical abnormalities. However the definite cause of this event is unclear. We conclude that it is difficult to predict a glaucoma attack following surgery under general anesthesia, but this complication is an important ophthalmologic emergency. Immediate diagnosis and appropriate treatment should be done to prevent the grave prognosis.

In an X-linked immunodeficiency with hyper IgM (X-HIM), several mutations of genes coding for CD40 ligand are responsible for B-cell defects. However, it is controversial whether the defective class switching from IgM to IgG or IgA but not IgE is rescued by in vitro stimulations or not. In six X-HIM patients, signaling through CD40 induced IgE but not IgG or IgA secretion in vitro. When transferred into SCID mice, however, B-cells of X-HIM patients secreted as much IgG and IgA as those of healthy controls. Moreover, sIgG and sIgA expressions were also induced by the in vivo stimulation. These results support that B-cells of X-HIM patients are defective of in vitro class switching from IgM to IgG or IgA and show that the in vivo stimulations rescue the defective class switching and suggest that the class switching of IgE and that of IgG or IgA might be regulated quite differently.

We examined the capacity of interleukin-4 (IL) production from lymphocytes and basophils, isolated from the peripheral blood of allergic patients sensitive to house dust mite, after stimulation with mite extract. IL-4 production was measured by a sensitive bioassay based on coculture with CT.h4S (a human IL-4-responsive cell line). Lymphocytes and basophils from patients with elevated serum IgE specific to mite allergen [radioallergosorbent test (RAST) score > 3] could produce detectable levels of IL-4 in response to mite extract, whereas those from patients with a RAST score of less than 2 or normal volunteers could not. The sensitivity of basophils to mite extract was high, so that a lower concentration of mite extract (1-10 ng/ml) could induce maximal IL-4 production. On the other hand, a higher concentration (10 micrograms/ml) was required for maximal IL-4 production from the lymphocytes. These findings demonstrate that allergen-specific IL-4-producing cells, lymphocytes and basophils, are generated in vivo in allergic patients and also that there exist characteristic differences between lymphocytes and basophils related to the in vivo source of IL-4.

We examined the capacity of T cells from normal individuals and allergic patients with very high IgE to differentiate into interleukin-4(IL-4)-producing cells in vitro. T cells incubated with anti-CD3 monoclonal antibody plus IL-4 or plus anti-IL-4 antibody in the presence of antigen-presenting cells for 7 days were restimulated and their capacity to express IL-4 mRNA was examined by RT-PCR. In T cells from normal individuals, there was a marked increase in the expression of IL-4 mRNA following the addition of IL-4. After fractionation of normal T cells into naive T (CD45RA+) and memory T (CD45RO+) cells, induction of the increase of IL-4 mRNA was restricted to the naive T cell population. In contrast, in T cells from allergic patients, the stimulation of whole or naive T cells with anti-CD 3 monoclonal antibody in the presence of IL-4 induced much less IL-4 mRNA. These findings suggest the presence of a functional abnormality in IL-4-dependent development of IL-4-producing T cells in the peripheral-blood naive T cells from allergic patients.

IgE synthesis by purified human B cells is induced by two signals: a class switching factor, most commonly interleukin (IL)-4, and the engagement of CD40, which is activated through its interaction with CD40 ligand (CD40L) expressed on activated T cells. Thus, the combination of IL-4 and anti-CD40 monoclonal antibody (mAb) has been shown to stimulate IgE production in vitro by highly purified B cells. In this T cell-independent system, strong homotypic aggregation of B cells is observed prior to the production of IgE. Flow cytometric analysis and cell binding assays showed that the stimulation of purified B cells with anti-CD40 mAb plus IL-4 resulted in a striking increase of intercellular adhesion molecule (ICAM)-1(CD54) expression, an induction of CD43 and an avidity change of lymphocyte functional antigen (LFA)-1(CD11a/CD18), with little augmentation of CD18 expression. Addition of anti-ICAM-1 mAb caused an inhibition of homotypic aggregation but augmented IgE synthesis by B cells stimulated with anti-CD40 mAb and IL-4, although it did not affect B cell proliferation or IL-6 production by the B cells. Among the mAb against counter-receptors for ICAM-1 tested, anti-CD11a mAb suppressed IgE synthesis, while anti-CD18 mAb and anti-CD43 mAb had little effect. The enhancing or inhibitory effect of anti-ICAM-1 mAb or anti-CD11a mAb on IgE production was achieved by the increased or decreased expression of germline C epsilon transcripts by B cells stimulated with anti-CD40 mAb and IL-4. These results indicate that B cell-B cell interaction through ICAM-1 and one of its counter receptors, LFA-1, regulates IgE synthesis by modulating C epsilon germ-line transcription.

The case of a 9-year-old girl with pulmonary stenosis complicated by acquired bleeding tendency after Jatene's operation is reported. Coagulation study revealed that platelet count and von Willebrand factor were reduced. Catheterization study revealed severe pulmonary stenosis, the pressure gradient between the right ventricle (RV) and pulmonary artery (PA) being 190 mmHg. A link between pulmonary stenosis and bleeding tendency was suggested and the patient was diagnosed as having acquired von Willebrand disease due to activated platelet/von Willebrand factor interactions enhanced by "shear stress" at the site of pulmonary stenosis. Right ventricular outflow reconstruction was done when she was 9 years-old. Due to a protocol based on the results of the challenge test that we instituted to determine the efficacy and effective duration of blood derivatives and hemostatic agents, perioperative massive bleeding was avoided. Bleeding tendency disappeared and coagulation study findings normalized with correction of the abnormal hemodynamic state.

Case 1: 55-year-old male was admitted to our hospital for further examination of increased transparency of X-ray in the left lower lung. He had history of pneumonia in his childhood. Left bronchography revealed mild cylindrical bronchi-ectasia in the proximal bronchi but poor filling by contrast in the peripheral bronchi. Case 2: 61-year-old male was referred to our hospital with palpitation and dyspnea. Chest X-ray film revealed hyperlucency of the left lower lung. The ventilation scan showed a marked decrease in ventilation to the left lung and air trapping was present in the left lung. The perfusion scan showed a matched decrease in blood supply to the left lung. In both cases, pulmonary arteriogram showed narrowed and withered-tree-like left pulmonary arteries. From these clinical findings, we diagnosed these two cases as Swyer-James syndrome. We are reporting two cases of Swyer-James syndrome and describing the clinical feature, differential diagnosis and etiology of the syndrome.

Ischemic changes in the left ventricular wall were investigated during graded coronary constriction in 9 dogs. Epicardial and intramyocardial electrograms were recorded and changes in ST-segment voltage were correlated with the changes in intramyocardial gas tensions in outer and inner layers of the myocardium measured using a mass spectrometer. Electrographic and metabolic changes appeared first in inner layer and proceeded to outer layer with increasing severity of coronary constriction. In both layers intramyocardial ST-segment elevation was closely related with the increase in carbon dioxide tension (r = 0.78 in inner layer and r = 0.67 in outer layer). Epicardial electrogram, however, reflected the regional ischemia less sensitively than intramyocardial electrograms and has a limited function to detect myocardial ischemia.

The purposes of this paper are, 1) to determine the dimensions representing the structure of conscious level "sex-role-acceptance" by the female adolescents in relation to the environments within which the subjects were brought up and educated, and 2) to investigate several factors under which the conscious structure of role-acceptance have been deeply influenced: i.e., parents, sibling, boy friends in intimate terms, and others (male or female). Our data were collected from more than 1800 female students, and were subjected to multivariate analyses. The results were: 1) The following three dimensions pertaining to their sex-role-related decision-making were obtained: Biological, Social-motivational, and Self-interest. 2) The difference in the conscious-structure between the role acceptance group and the non-acceptance group was statistically significant. 3) the father-daughter interaction was significantly important in creating an active contributional will in the girls' life-plan, while the mother-daughter interaction turned out to have a rather negative influence upon their will of contribution. 4) The lack of identification with parents was sometimes compensated by their identification with others. The social importance of positive, warm support for the youngsters was clearly demonstrated.

Effect of coronary ligation on epicardial and intramyocardial ST-segment voltage was investigated in 121 dogs. After ligation of the anterior descending coronary artery, marked ST-elevation, more than 2 mV, was detected in 102 dogs (Group-I) in epicardial lead of the area which was nourished by the ligated coronary artery. In 7 dogs (Group-II), coronary ligation produced negligible changes in ST-segment voltage in epicardial lead, while significant ST-elevation was observed in the leads of middle and inner layers of the myocardium. Systemic hemo-dynamic parameters showed no statistically significant difference between the 2 groups. Coronary collateral indices, however, revealed the significant difference between them. Systolic peripheral coronary pressure was 29.3 +/- 3.1 mmHg in Group-I and 59.2 +/- 10.6 mmHg in Group-II (P less than 0.05). Retrograde flow was also greater in Group-II (14.8 +/- 4.4 ml/min) than in Group-I (1.3 +/- 0.5 ml/min) (p less than 0.01). Diversion of retrograde flow produced a marked ST-elevation in Group-II. Restoration of retrograde flow to the myocardium immediately decreased the ST-segment voltage. These results suggest that spontaneously developed collaterals have an important function for the prevention of the occurrence of ischemic injury after coronary ligation in dogs.

This study was undertaken to investigate the effects of graded coronary constriction on regional gas tensions of the myocardium. In 12 open chest dogs, tissue carbon dioxide (PtCO2) and oxygen (PtO2) tensions were measured simultaneously in outer and inner layers of the myocardium using a mass spectrometer. In normal condition, higher PtO2 and lower PtCO2 were observed in outer layer than in inner layer. With application of coronary constriction, increase in PtCO2 and decrease in PtO2 were observed in both layers of the myocardium, but the response to the ischemic stimuli by applying coronary constriction in inner layer was different from that in outer layer. Severe coronary constriction, more than 90% in its diameter, was necessary to produce significant changes in both gas tensions in both layers of the myocardium. Decrease in PtO2 was found in the condition of less severe coronary constriction and to be greater in inner layer than in outer layer of the myocardium. In terms of the changes in PtCO2, inner layer was also more susceptible to the ischemic stimuli than outer layer. The greater and earlier elevation of PtCO2 in inner layer than in outer layer is regarded as one of the possible mechanisms of the reduction of myocardial contraction in the early stage of myocardial ischemia.

The functional capacity of coronary collaterals was investigated under resting and stress conditions in 15 dogs with chronic occlusion of the anterior descending coronary artery. ST-elevation in epicardial and intramyocardial electrograms was used for assessing the degree of regional myocardial ischemia. In resting condition, epicardial and intramyocardial electrograms from collateral, intermediate and circumflex areas revealed no significant ischemic changes. In 6 of 15 dogs in which atrial pacing was performed, ST-elevation appeared in electrograms from collateral and intermediate areas. In another 9 dogs, pacing tachycardia produced no significant changes in all electrograms. In these dogs more than 50% constriction of the donor coronary artery was necessary to induce ST-elevation in electrogram from collateral area. These findings indicate that the coronary collateral vessels in dog play beneficial role to prevent the myocardium from ischemic injury under stress condition, although their functional capacity is not comparable to that of normal coronary artery.