Association study between kynurenine 3-monooxygenase gene and schizophrenia in the Japanese population.

Several lines of evidence suggest that metabolic changes in the kynurenic acid (KYNA) pathway are related to the etiology of schizophrenia. The inhibitor of kynurenine 3-monooxygenase (KMO) is known to increase KYNA levels, and the KMO gene is located in the chromosome region associated with schizophrenia, 1q42-q44. Single-marker and haplotype analyses for 6-tag single nucleotide polymorphisms (SNPs) of KMO were performed (cases = 465, controls = 440). Significant association of rs2275163 with schizophrenia was observed by single-marker comparisons (P = 0.032) and haplotype analysis including this SNP (P = 0.0049). Significant association of rs2275163 and haplotype was not replicated using a second, independent set of samples (cases = 480, controls = 448) (P = 0.706 and P = 0.689, respectively). These results suggest that the KMO is unlikely to be related to the development of schizophrenia in Japanese.

Selenium deficiency: report of a case.

We report an 18-month-old Japanese boy with selenium deficiency. He had dry skin with irregularly shaped, erythematous changes on the cheeks, groin, hip, and extremities, erosions on the external urethral and anal orifices, and sparse, short, thin, light-coloured hair. He had received parenteral nutrition for 5 months because of juvenile polyposis. At presentation, his serum selenium level was less than 2.0 microg/dL (normal range, 10.6-17.4 microg/dL). His skin lesions responded well to supplementary treatment with sodium selenite. His skin symptoms were similar to those attributable to a deficiency of zinc which, like selenium, is an essential trace element. According to the literature, selenium deficiency is responsible for cardiomyopathy, which was diagnosed in our patient. The clinical similarity to zinc deficiency and the literature yielded important clues for a diagnosis of selenium deficiency in this patient.

CD14-mediated induction of interleukin-8 and monocyte chemoattractant protein-1 by a heat-resistant constituent of Porphyromonas gingivalis in endothelial cells.

Viable and inactivated Porphyromonas gingivalis dose-dependently induced interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) secretion in human umbilical vein endothelial cells (HUVECs). The inactivated P. gingivalis, in comparison with viable bacteria, tended to enhance the production of both chemokines more strongly. The production of MCP-1 protein began increasing immediately after stimulation by P. gingivalis, and there was a nearly linear increase from 0 to 8 h of incubation, whereas IL-8 production showed a linear increase between 4 and 12 h of incubation. The IL-8 and MCP-1 mRNA expressions in HUVECs as determined by reverse transcriptase-polymerase chain reaction (RT-PCR) or Quantikine mRNA colorimetric quantification kits were found to be enhanced by P. gingivalis. Furthermore, the time courses of IL-8 and MCP-1 mRNA expressions were in accordance with those of protein production. Addition of polymyxin B or boiling did not weaken the stimulatory effect of P. gingivalis, which inhibited the effect of Escherichia coli lipopolysaccharide (E. coli LPS) and tumour necrosis factor-alpha (TNF-alpha), respectively. In contrast, the induction of IL-8 and MCP-1 by P. gingivalis was significantly reduced by anti-CD14 antibody. Our results suggest that some heat-stable component of P. gingivalis, including LPS, may be responsible for the induction of IL-8 and MCP-1 in HUVECs by a CD14-dependent mechanism. These effects might be involved in the accumulation and activation of neutrophils and monocytes at an early stage of the periodontal pathogenesis.

A heat-stable component of Bartonella henselae upregulates intercellular adhesion molecule-1 expression on vascular endothelial cells.

Bartonella henselae upregulated the expression of intercellular adhesion molecule-1 (ICAM-1) on human umbilical vein endothelial cells (HUVECs). The induction level of ICAM-1 depended on the inoculation bacterial dose. ICAM-1 expression began increasing 4 h after infection and reached a sustained peak beginning at 12 h after B. henselae infection; this time course was similar to that of lipopolysaccharide (LPS) of Escherichia coli. The stimulatory effect was abolished when live B. henselae were separated from HUVECs by a filter membrane. The nonpiliated strain, which is unable to invade endothelial cells, induced ICAM-1 expression to the same extent as the piliated strain. Inactivation of B. henselae by ultraviolet (UV) irradiation, heat (56 degrees C, 30 min), or sonication did not alter its stimulatory activity. Polymyxin B, which strongly inhibited the effect of LPS, did not exert any influence on the stimulatory activity of B. henselae. Furthermore, the effect of sonicated B. henselae was not inhibited even by boiling, which was also the case with LPS. Our data suggest that some heat-stable component of B. henselae binds to the endothelial cell surface, inducing ICAM-1 expression. Though the participation of LPS could not be completely ruled out, we suppose that some unidentified heat-stable proteins, lipids, or polysaccharides may be the stimulatory factor(s). The ability of B. henselae to enhance the expression of adhesion molecules on endothelial cells may be an important mechanism in the pathogenesis of B. henselae infection.

[A case of primary Sjögren syndrome with repeated purpura].

In Sjögren syndrome, purpura is one of its various well known eruptions. Although this disease state is assumed to be based on hypergammaglobulinemia, the details of its mechanism are unknown. We experienced a case involving a female patient with primary Sjögren syndrome showing repeated purpura on the legs, and examined her blood viscosity and histopathology. This girl developed Sjögren syndrome and was admitted to our hospital at 12-years-old. She underwent steroid treatment because of aggravation of the xerosis state and prominent purpura on the legs. Hypergammaglobulinemia was improved during the course; however, purpura appeared repeatedly. Although her blood viscosity was slightly higher than normal, this had no relation to purpura and serum gamma globulin values. Skin biopsy revealed necrotizing angiitis. These results suggest that the purpura of this case was caused not only by hyperviscosity from the hypergammaglobulinemia but also involvement of vasculitis by the primary disease.

Anti-alpha-fodrin antibodies in Sjögren's syndrome in children.

OBJECTIVE: To investigate the prevalence of anti-alpha-fodrin antibody specific for adult Sjögren's syndrome (SS) in patients with juvenile onset SS. METHODS: Serum anti-alpha-fodrin antibody was examined in 15 patients with juvenile SS (11 cases of primary SS and 4 secondary SS) and in 16 children with systemic lupus erythematosus (SLE) by Western blot analysis using a recombinant 120 kDa alpha-fodrin fusion protein. RESULTS: All the 15 serum samples from patients with SS reacted with a recombinant alpha-fodrin fusion protein in Western blot analysis. In contrast, reactivity was found in only 2 of the 16 patients with SLE. The clinical features of the 15 patients with juvenile onset SS were very specific; only 4 patients complained of dryness, while 6 had abnormal excretion ability. Salivary gland enlargement was the most common clinical manifestation. Characteristic laboratory findings in juvenile onset SS included a higher prevalence of antinuclear antibodies, anti-SSA/Ro antibodies, and rheumatoid factor, as well as increased erythrocyte sedimentation rate and hypergammaglobulinemia. CONCLUSION: The pathogenesis of juvenile SS seems to be the same as that of adult SS, although subjective symptoms of dryness are less frequent in juvenile cases. This discrepancy may indicate that SS is a slowly progressive disease with a long time span. The anti-alpha-fodrin antibody is likely to be a reliable diagnostic marker for juvenile SS.

Live Bartonella henselae enhances endothelial cell proliferation without direct contact.

The proliferation of human umbilical vein endothelial cells (HUVECs) cocultivated with live B. henselae was enhanced in a bacterial dose-dependent manner, and the stimulatory effect was specific to vascular endothelial cells. The inactivation of B. henselae by UV or heat treatment abolished its stimulatory activity, suggesting that live bacteria is necessary for the growth stimulation effect. To investigate the role of direct contact, live B. henselae were separated from HUVECs by a filter membrane (Millicell-CM insert). Even under this condition, an enhanced proliferation of HUVECs was observed. However, no morphological changes in the HUVECs were apparent compared to the B. henselae -infected cells. Furthermore, we isolated a nonpiliated strain of B. henselae that is unable to attach to and enter into endothelial cells. The nonpiliated strain possessed the ability to stimulate the proliferation of cocultivated HUVECs the same as the piliated strain. Moreover, the culture supernatants of B. henselae were also able to induce HUVEC proliferation. Our results indicate that the stimulation of HUVEC proliferation by B. henselae is mediated by soluble factor(s) secreted from the bacteria.

Increased circulating vascular endothelial growth factor is correlated with disease activity in polyarticular juvenile rheumatoid arthritis.

OBJECTIVE: To investigate the relevance of vascular endothelial growth factor (VEGF) in the pathogenesis of juvenile rheumatoid arthritis (JRA). METHODS: Serum VEGF levels in 58 patients with JRA (systemic in 17, polyarticular in 29, pauciarticular in 12) were measured by ELISA and compared with those of 21 patients with infectious diseases and 50 healthy children. Correlations of VEGF levels with number of joints with active arthritis, erythrocyte sedimentation rate (ESR), and hyaluronic acid (HA) were examined. RESULTS: Serum levels of VEGF in patients with JRA were significantly higher than in healthy controls. Patients with systemic and polyarticular JRA showed statistically higher levels of VEGF than those with infectious diseases. VEGF levels correlated statistically with C-reactive protein (CRP) in patients with both infectious diseases and polyarticular JRA, but the regression slope (VEGF/CRP) was much steeper in polyarticular JRA than in infectious diseases. Serum VEGF levels correlated with disease activity variables such as the number of joints with active arthritis, ESR, and serum HA levels in polyarticular JRA. CONCLUSION: The correlation of serum VEGF levels and disease activity in polyarticular JRA suggests that VEGF may take an active part in joint inflammation.

[A case report of systemic lupus erythematosus patient with severe lupus retinopathy and antiphospholipid antibody].

An eleven-year-old boy with systemic lupus erythematosus (SLE) developed severe bilateral lupus retinopathy when he was in active stage of SLE. The patient, who had suffered from SLE for 3 years, was admitted to our hospital because of high grade fever, systemic lymphadenopathy, leukopenia, elevation of erythrocyte sedimentation rate and hypocomplementemia. The dose of prednisolone was increased considering he was exacerbated of SLE, however, the convulsion as CNS lupus occurred to him. After the event he noted loss of vision in his bilateral eyes. The ophthalmologic examination revealed the lesions of cotton-wool spots, retinal vessel dilatations and diffuse occlusions of the retinal arterioles and venules which were compatible with lupus retinopathy. Although the coagulation time was normal, antiphospholipid antibodies were positive at the time of ocular involvement. Careful attention should be paid to the occurrence of lupus retinopathy when the patients with SLE developed in the active stage or CNS lupus, especially they have antiphospholipid antibodies.

Expression of vascular endothelial growth factor in sera and lymph nodes of the plasma cell type of Castleman's disease.

To evaluate the possible involvement of vascular endothelial growth factor (VEGF) in the pathogenesis of Castleman's disease, we studied VEGF levels in sera and supernatants of cultured lymph nodes from two patients with the plasma cell type of Castleman's disease, and analysed the expression of VEGF immunohistochemically in the lymph nodes. Clinically, one patient was classified as the localized type and the other as the multicentric type. Histologically, mature plasma cells and hyalinized vessels were prominent in the interfollicular region. The VEGF levels of the sera and the supernatants of cultured lymph nodes of both patients were higher than those of normal controls. VEGF was strongly expressed in plasma cells in the interfollicular region of the lymph nodes of both patients, but rarely in normal lymph nodes. Our results suggest that VEGF may be involved in the marked vascular proliferation in the interfollicular region of the lymph nodes of the plasma cell type of Castleman's disease.

Induced production of nitric oxide and sensitivity of alveolar macrophages derived from mice with different sensitivity to Coxiella burnetii.

We compared in vitro sensitivities to Coxiella burnetii of alveolar macrophages, derived from mice sensitive and resistant to C. burnetii, respectively, and examined the role of nitric oxide (NO) in the C. burnetii infection. Alveolar macrophages of sensitive A/J mice showed a larger population of C. burnetii antigen-positive cells than those of resistant C57BL/6 mice. C. burnetii induced NO production in alveolar macrophages, but N-methyl-L-arginine and sodium nitroprusside (SNP), NO inhibitor and donor, respectively, did not inhibit the infection. Thus the NO induction seems to be independent of the cell defense mechanism against the C. burnetii infection.

Increased serum levels of vascular endothelial growth factor in Kawasaki disease.

Vascular endothelial growth factor (VEGF) is an angiogenic mitogen that specifically targets vascular endothelial cells. The objective of this study was to evaluate the role of VEGF in Kawasaki disease (KD), the most common cause of systemic vasculitis in childhood. Serum VEGF levels were measured by ELISA in 22 patients with KD, 22 febrile children with infection, and 19 healthy children. Samples from KD patients were divided into three groups: acute stage (n = 20), subacute stage (n = 13), and convalescent stage (n = 15). The results showed that KD patients in the acute and subacute stages had significantly higher levels of VEGF than did patients with infectious diseases and the healthy control subjects. When compared with the VEGF levels of patients with and without coronary artery lesions (CAL), significantly higher levels of VEGF were observed in the subacute stage in patients with CAL and in patients without CAL in the acute stage. Serial examination revealed that the serum VEGF levels in KD patients with CAL increased from a relatively low level in the acute stage to an extremely high level in the subacute stage. In contrast, patients without CAL were found to have extremely high levels of VEGF only in the acute stage of KD. In KD patients, the serum VEGF levels did not correlate with the inflammatory markers and clinical symptoms. Our results raise the possibility that VEGF is involved in the pathogenesis of KD, especially in the development of CAL. Further study is needed to clarify the biologic effect of VEGF on coronary arteries in KD.

[Two cases with SLE and MCTD developed after a long period of chronic arthritis that was initially diagnosed as JRA].

In order to discuss the diversity of clinical features and the difficulty in diagnosis of children with juvenile rheumatoid arthritis (JRA), we present two cases who have documented the development of systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD) after a long period of disease characterized only by arthritis that was initially diagnosed as JRA. The first case was a girl diagnosed for her arthritic joints as polyarticular JRA at 15 years of age. At onset, she had Raynaud phenomenon but autoantibodies such as anti-nuclear antibody (ANA), anti-DNA antibody, and rheumatoid factor were negative. Five years after onset, she became ANA positive and 3 years later she became pregnant. During her pregnancy, she became positive for anti-DNA antibody without any signs of nephritis. One month after the delivery, however, she developed butterfly rash, carditis, nephritis, and was diagnosed as SLE. No destructive changes were observed in her joints though arthritis continued for 8 years form onset to pregnancy. The second case was a 3 years old girl who was diagnosed as polyarticular JRA. Treatment by aspirin induced complate remission after one year from the onset. However, 10 years after that remission, she developed Raynaud phenomenon and arthralgia in her knees and hip joints. Her laboratory findings showed hypergammaglobulinemia, positive ANA, positive anti-DNA antibody, positive anti-RNP antibody. She was eventually diagnosed as MCTD when she was found to have polymyositis by EMG and serum CK. In the present paper, two cases imply the difficulty in diagnosing JRA and diversity of rheumatic diseases such as JRA, SLE and MCTD. Closer and longer period of observation is essential for the JRA patients with nondestructive arthritis.

Clinical features of Japanese children and adolescents with systemic lupus erythematosus: results of 1980-1994 survey.

Marked advances have been made in the past decade in the management of adults with systemic lupus erythematosus (SLE). Therefore, a nationwide retrospective survey was conducted between 1980 and 1994 to investigate the clinical manifestations of SLE in Japanese children and adolescents. Questionnaires were sent to 340 hospitals. Of 405 patients reported by 176 hospitals, 373 patients, diagnosed by the criteria established by the Pediatric Study Group of the Japanese Ministry of Health and Welfare in 1985, were enrolled in the study. Forty-nine of the 354 patients (13.8%) had relatives with a connective tissue disease within the third degree of consanguinity. The frequent manifestations in 373 patients were the presence of antinuclear antibody (98.9%), immunologic disorders (93.0%), hypocomplementemia (87.1%), malar rash (79.6%) and fever (74.0%). Lupus nephritis was present in 148 of the 309 patients (47.9%) at their first visit to a clinic, and 261 of the 373 patients (70.0%) developed renal involvement during the observation period. Of 370 patients, 92 patients (24.9%) exhibited central nervous system lupus. Of 368 patients, 192 patients (52.2%) were treated by methylprednisolone pulse therapy and 148 patients (40.2%) received immunosuppressants in combination with steroid therapy at some stage during the observation period, Survival rate at 5 years from onset was 95.9%. Management of infection, coagulopathies, and central nervous system involvement is essential to improve the prognosis of SLE in Japanese children and adolescents.

Serum levels of hyaluronic acid indicate the severity of joint symptoms in patients with systemic and polyarticular juvenile rheumatoid arthritis.

OBJECTIVE: Elevated serum levels of hyaluronic acid (HA) correlate with joint inflammation in adult rheumatoid arthritis (RA). There are no laboratory indices for specifically assessing joint inflammation. Therefore, serial measurements of HA were assessed as a possible tool for measuring the severity of arthritic symptoms in children with juvenile rheumatoid arthritis (JRA). METHODS: Serum levels of HA, measured by a sandwich assay method using HA binding protein, were correlated with the severity of joint symptoms and with laboratory test values in 71 patients with JRA, 30 children with other rheumatic diseases, and 138 children without rheumatic disease. RESULTS: Serum levels of HA showed significant correlation with the severity of joint symptoms, but not with systemic symptoms, in patients with systemic and polyarticular JRA. No other laboratory tests, including C-reactive protein and erythrocyte sedimentation rate, reflected the severity of joint symptoms. This correlation of serum levels of HA with joint symptoms was observed in patients with systemic and polyarticular JRA, but not in pauciarticular JRA, other rheumatic diseases, or nonrheumatic diseases, even when signs of arthritis were present in the latter 3 groups. CONCLUSION: Serum levels of HA are useful in objectively evaluating arthritic symptoms in patients with systemic and polyarticular JRA, and may have diagnostic value in this disease.

[Clinical features of infantile patient with juvenile rheumatoid arthritis].

Little is known about clinical features of infantile juvenile rheumatoid arthritis (JRA) because it is very rare for the patients to develop JRA within one year of age. In the past 20 years, we experienced three JRA patients whose onset was under 1 year of age. The incidence of infantile JRA was 3.2% of all JRA patients in our facility. They are 9 month-old male with systemic onset, 6-month-old female with polyarticular onset and 8 month-old female with systemic onset. It was difficult to evaluate subjective symptoms such as arthralgia or morning stiffness since the patients could not complain precisely. Therefore, careful observation on their behaviors, such as the delayed development of their motor function and bad humor and/or loss of activity in the morning, was important for evaluating joint symptoms. In case 1, measuring the serum level of hyaluronic acid was specifically useful to evaluate the arthritis. Drug therapy was not successful especially in infantile JRA. One of the reason for this ineffectiveness of drug therapy might be explained by poor adsorption of drugs in infants; the serum acetyl salicylic acid level was lower in infantile patients than the other patients with JRA even though they received enough dose of aspirin. Infantile JRA was revealed to have specific difficulties in early diagnosis and adequate treatment. Therefore, accumulated case studies about clinical features of infantile JRA is essential for their better prognosis.

Adrenergic mechanisms associated with the movement of platelets in iridophores from the freshwater goby, Odontobutis obscura.

1. Cultured iridophores from the freshwater goby, Odontobutis obscura, were used to investigate adrenergic mechanisms of movement of platelets in the iridophores. 2. Norepinephrine, which was assumed to be the transmitter of the iridophore nerves, induced dispersion of platelets within the cells. 3. The effect of norepinephrine was inhibited by an alpha-adrenergic antagonist, yohimbine, but not by a beta-adrenergic antagonist, propranolol. 4. Isoproterenol, a beta-adrenergic agonist, failed to bring about aggregation of platelets. 5. Forskolin, an activator of adenylate cyclase, was effective in inducing aggregation of platelets. 6. 8-Br-cAMP caused the aggregation of platelets and inhibited the norepinephrine-induced dispersion of platelets. 7. It appears that the adrenoceptors of the iridophores of this species are solely of the alpha type; they mediate the dispersion of platelets; and an increase in intracellular levels of cAMP induces the aggregation of platelets.