MEFV mutation analysis of familial Mediterranean fever in Japan.

BACKGROUND: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent attacks of fever with serosal inflammation. FMF gene (MEFV) mutations have been identified primarily in patients from Mediterranean populations. Although several clinical cases have been reported in Japan, there have been few reports to date on mutation analysis. We studied FMF patients and their relatives to examine the clinical and genetic features of this disease in the Japanese population. METHODS: Twelve Japanese FMF patients who met the Tel Hashomer criteria and a total of 17 relatives from 5 of 10 families underwent molecular genetic studies to detect MEFV mutations. The characteristics of these Japanese FMF patients and geno-phenotypical correlations were examined. RESULTS: Almost all of our patients had been suffering for a long time from fever of unknown origin and one patient also had systemic amyloidosis. In our 12 FMF patients, we detected the substitutions E84K, L110P, E148Q, R761H and M694I. We also newly diagnosed 2 relatives as having FMF based on clinical symptoms and the existence of FMF mutations. One patient was homozygous for E148Q, the patient with systemic amyloidosis was a homozygote for M694I and 4 patients from 3 families were compound heterozygotes for E148Q and M694I. Three patients in one family were compound heterozygotes for E148Q, L110P and M694I. There were 3 patients who were heterozygous for E84K, L110P-E148Q or M694I and had no other nucleotide changes in the exons of MEFV. On the other hand, 2 relatives who had never experienced symptoms of FMF were homozygous for L110P-E148Q as well as compound heterozygous for E148Q/E148Q-R761H. E148Q and M694I were the most frequently detected substitutions in our study. CONCLUSIONS: MEFV mutations occur in Japanese FMF patients though FMF is rare in Japan. The identification of MEFV mutations could be a reliable diagnostic test for FMF. The results of genetic analyses on 14 Japanese FMF patients in this study revealed that E148Q and M694I are frequent alleles.

Tec protein-tyrosine kinase is involved in the thrombopoietin/c-Mpl signaling pathway.

Thrombopoietin (TPO) is the ligand of the proto-oncogene product, c-Mpl, and supports the growth and maturation of megakaryocytic cells. Although much attention has been paid to the in vivo role of the TPO/c-Mpl system in the context of maintenance of circulating platelets, still little is understood about how activation of c-Mpl leads to mitogenesis and differentiation inside cells. Tec protein-tyrosine kinase (PTK) is the prototype of a recently emerging subfamily of nonreceptor type PTKs. Tec has been demonstrated to be activated by a wide range of cytokine stimulations. In this paper we show that TPO stimulation also rapidly enhances tyrosine-phosphorylation and activity of the Tec kinase in a human TPO-dependent cell line. In addition, the Vav protein, a blood cell-specific signaling molecule, is shown to be tyrosine-phosphorylated in response to TPO and to be constitutively associated with the Tec protein. From this evidence, we conclude that Tec is involved in the intracellular signaling system of TPO/c-Mpl.

Reduced host resistance and Th1 response to Cryptococcus neoformans in interleukin-18 deficient mice.

Using interleukin (IL)-18 deficient (IL-18(-/-)) mice, we examined the role of IL-18 in the host resistance and Th1 response against infection with Cryptococcus neoformans. Fungal clearance in the lung was reduced in IL-18(-/-) mice, although there was no significant change in the level of dissemination to the brain. The DTH response, as determined by footpad swelling, was also diminished in IL-18(-/-) mice compared to control wild-type (WT) mice. The levels of IL-12 and interferon (IFN)-gamma in the sera were significantly lower in IL-18(-/-) mice than in WT mice. Spleen cells from infected WT mice produced a high level of IFN-gamma upon stimulation with the microbe, while only a low level of IFN-gamma production was detected in spleen cells from infected IL-18(-/-) mice. Administration of IL-18 almost completely restored the reduced response in IL-18(-/-) mice, while IL-12 showed a marginal effect. These results demonstrated the important role of IL-18 in the resistance and Th1 response of mice to C. neoformans by potentiating the production of IFN-gamma.

Herpes simplex virus bronchopneumonia in a non-immunocompromized individual.

We report a rare case of herpes simplex virus (HSV) bronchopneumonia in an otherwise healthy middle-aged individual. Bronchoscopy indicated scattered white-coated lesions in the bronchial mucosa. The diagnosis was established following immunohistopathological staining for HSV of specimens obtained by bronchial biopsy. This case suggests that HSV could be a pathological agent of not only oral and genital mucosal lesions but also lower respiratory tract infection.

[Hemorrhagic myocardial infarct. An analysis of the distribution of the hemorrhage and of the permeability of the artery related to the infarct]. / Infarto hemorrágico do miocárdio. Análise da distribuição da hemorragia e da perviabilidade da artéria relacionada ao infarto.

PURPOSE: To study the extent of hemorrhagic myocardial infarction (HMI) and patency of the infarct related artery. METHODS: Forty seven cases of HMI diagnosed by necropsy (patient age range 30-81 years, mean 59) were studied retrospectively. Hemorrhagic extent was evaluated by microscopic analysis of myocardial sections of the infarcted areas and coronary patency was studied by angiography and by serial coronary sections at necropsy. RESULTS: In 12 cases hemorrhage extended outside the infarcted area and in the remaining cases it was restricted to the necrotic zone. Coronary patency was spontaneous in 8 of 24 cases, secondary to thrombolytic therapy or angioplasty in 8 and post coronary artery bypass in 15. Recent occlusive thrombus was diagnosed in 26 of 44 cases. Grouping all cases according to angiographic or macro and microscopic evidences of coronary patency, it was found that 35 of 47 studies cases (74.4%) had the infarct related coronary artery free of occlusion. In most cases of HMI myocardial hemorrhage restricted to the infarcted necrotic zone but in almost 25% it could reach areas beyond the infarcted necrotic zone probably resulting in deleterious consequences. CONCLUSION: Reperfusion is frequent and it plays a role in the hemorrhagic event but it was not seen in 25% of these studied cases. These findings suggest that other mechanisms could participate of the pathogenesis of HMI.

[Retrospective analysis of elderly patients > or = 60 years of age with acute leukemia].

A retrospective analysis was performed on 76 consecutive elderly patients with acute leukemia aged 60 years or more (48 men, 28 women). Forty patients were 60-69 years old, 28 were 70-79 years old and 8 were > or = 80 years old. There were 55 patients with acute myelogenous leukemia (AML), 13 acute lymphoblastic leukemia (ALL) and 8 AML from myelodysplastic syndrome (MDS/AML). Patients were treated with the JALSG protocol, CAG regimen, or low-dose Ara-C regimen for AML, and DVP/M-CHOP protocol for ALL. The complete remission (CR) rates were 52.7% (29 of 55) in AML, 61.5% (8 of 13) in ALL, and 0% in MDS/AML. The median CR durations were 226, 85, 0 days, and the median survivals were 204, 177, 99 days, respectively. CR rates were 65.3% for the JALSG protocol, 62.5% for the CAG regimen and 25.0% for low-dose Ara-C regimen. According to age, CR was obtained 62.5% in patients aged 60-69 years and 33.3% in patients over 70 years old. Our results indicated that patients aged 60-69 years should be treated with intensive chemotherapy.

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

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

[How to set up a sleep laboratory in a community general hospital].

In Japan, sleep-related disorders (SRD) have been handled by only a few university teams, and mainly on a research basis, because polysomnography (PSG) is labor-intensive, expensive, and unprofitable. Although public consciousness of SRDs has been raised by mass-media campaigns, the situation is far from ore that would meet patient's demands. We have done 1476 PSG tests over about five years, and diagnosed more than 600 cases of SRDs. Therapy included nasal continuous positive airway pressure in 170 cases, prosthetic mandibular advancement in 55 cases, and uvulopalatopharyngoplasty and related operations in 97 cases. We describe how we set up a sleep laboratory and a care system, and how to circumvent problems. Several points related to the selection of patients for PSG in daily clinical practice can be clarified with our data. Computerized sleep respiratory analyser might save time and money, but their advantages and disadvantages should be studied. The approach to SRDs should be comprehensive and systematic, and polysomnographers play an important role. We need programs organized by scientific institutions to train polysomnographers. Research should continue not only on scientific aspects of SRDs but also on the way patients with SRDs are cared for. Reasonable insurance coverage of the costs of nosal continuous positive airway pressure and of PSGs is essential for widespread access to these services.

Chemokine synthesis and cellular inflammatory changes in lungs of mice bearing p40tax of human T-lymphotropic virus type 1.

To elucidate the pathogenic mechanisms of human T-lymphotropic virus type 1 (HTLV-1)-associated lung inflammation, we conducted a histopathological and molecular analysis study using transgenic mice bearing pX region of this virus. In these mice, accumulations of inflammatory cells consisting mainly of lymphocytes were present in peribronchiolar and perivascular areas and alveolar septa, while control littermate mice did not show such changes. In situ hybridization showed that the anatomic distribution of p40tax mRNA was similar to that of inflammatory cells, typically in peribronchiolar areas and to a lesser extent in perivascular and alveolar septa. Inflammatory cytokines, including IL-1beta, tumour necrosis factor-alpha and interferon-gamma, and several chemokines, such as monocyte chemotactic protein-1 (MCP-1), RANTES, macrophage inflammatory protein-1alpha (MIP-1alpha) and IP-10, were detected in lungs of transgenic mice but not control mice. Semiquantitative analysis using reverse transcription-polymerase chain reaction showed a significant correlation between MCP-1 mRNA expression and p40tax mRNA, but not with other chemokines. The gene expression of the above chemokines, with the exception of MIP-1alpha, correlated with the severity of histopathological changes in the lung. Considered together, our results suggested that p40tax synthesis may be involved in the development of lung lesions caused by HTLV-1 through the induction of local production of chemokines.

Induction of lymphocytic inflammatory changes in lung interstitium by human T lymphotropic virus type I.

Human T lymphotropic virus type I (HTLV-I) is the etiological agent of adult T cell leukemia, and reports suggest that several other clinical conditions are associated with HTLV-I infection, including myelopathy and inflammatory pulmonary diseases. However, the clinical entity of HTLV-I-associated lung disease remains unsubstantiated more than 10 years after its description. In the present study, we conducted a histopathological analysis of lung tissues of transgenic mice that expressed gene segments of HTLV-I p40(tax) regions. The aim of the study was to examine the relationship between expression of viral components and development of lung disorders. In these mice, inflammatory changes with infiltration of lymphocytes in peribronchial and perivascular areas and in alveolar septa developed at 11 wk of age and increased in incidence during the observation period (26 wk). There was a significant correlation between the pulmonary pathological changes and the level of expression of p40(tax) mRNA in the lungs. Our results provided for the first time strong evidence of a direct relationship between HTLV-I and development of bronchopulmonary infection.

Tec protein-tyrosine kinase is an effector molecule of Lyn protein-tyrosine kinase.

The Tec family is a recently emerging subfamily among nonreceptor type protein-tyrosine kinases (PTKs) consisting of Tec, Txk, Btk, Bmx, and Itk/Tsk/Emt. They have a long amino-terminal unique region containing a pleckstrin homology domain and a Tec-homology domain. We could previously show that, through the Tec-homology domain, Tec is bound to Lyn kinase both in vitro and in vivo. Because Tec is coexpressed with Lyn in many hematopoietic cell types, it has been intriguing to investigate the biological role of the Tec-Lyn association. Here we demonstrate that Lyn can phosphorylate tyrosine residues of the Tec protein, and thereby activate Tec in 3T3 fibroblasts. However, coexpression of Tec has little effect on the phospho-tyrosine-contents of Lyn. By using the in vitro kinase assay and the yeast system, we could prove that the Tec protein is a direct substrate of the Lyn kinase both in vitro and in vivo. From this evidence we conclude that Tec acts downstream of Lyn in intracellular signaling pathways. This is a novel case where one PTK is phosphorylated and regulated by another.

Tec protein tyrosine kinase is involved in the signaling mechanism of granulocyte colony-stimulating factor receptor.

Granulocyte colony-stimulating factor (G-CSF) is a critical cytokine to promote the growth, differentiation, and functional activation of myeloid cells. Despite its important roles, little is still understood how one cytokine can trigger such pleiotropic effects. By using mouse cell lines which can grow or differentiate in response to G-CSF, we investigated whether Tec protein tyrosine kinase is involved in either of the signaling pathways. Interestingly, Tec is shown to be tyrosine phosphorylated and activated by the G-CSF stimulation in both cell growth and differentiation mechanisms. Vav is also shown to be associated with Tec and tyrosine phosphorylated in response to G-CSF. Tec is a good candidate for protein-tyrosine kinases involved in both growth and differentiation mechanisms of myeloid cells.