Hidetoshi Ujiie

Adiponectin Acts as an Endogenous Antithrombotic Factor

Objective—Obesity is a common risk factor in insulin resistance and cardiovascular diseases. Although hypoadiponectinemia is associated with obesity-related metabolic and vascular diseases, the role of adiponectin in thrombosis remains elusive. Methods and Results—We investigated platelet thrombus formation in adiponectin knockout (APN-KO) male mice (8 to 12 weeks old) fed on a normal diet. There was no significant difference in platelet counts or coagulation parameters between wild-type (WT) and APN-KO mice. However, APN-KO mice showed an accelerated thrombus formation on carotid arterial injury with a He-Ne laser (total thrombus volume: 13.36±4.25×107 arbitrary units for APN-KO and 6.74±2.87×107 arbitrary units for WT; n=10; P<0.01). Adenovirus-mediated supplementation of adiponectin attenuated the enhanced thrombus formation. In vitro thrombus formation on a type I collagen at a shear rate of 250 s−1, as well as platelet aggregation induced by low concentrations of agonists, was enhanced in APN-KO mice, and recombinant adiponectin inhibited the enhanced platelet aggregation. In WT mice, adenovirus-mediated overexpression of adiponectin additionally attenuated thrombus formation. Conclusion—Adiponectin deficiency leads to enhanced thrombus formation and platelet aggregation. The present study reveals a new role of adiponectin as an endogenous antithrombotic factor.

Complement C1q regulates LPS-induced cytokine production in bone marrow-derived dendritic cells.

We show here that C1q suppresses IL‐12p40 production in LPS‐stimulated murine bone marrow‐derived dendritic cells (BMDC). Serum IL‐12p40 concentration of C1q‐deficient mice was higher than that of wild‐type mice after intraperitoneal LPS‐injection. Because neither globular head of C1q (gC1q) nor collagen‐like region of C1q (cC1q) failed to suppress LPS‐induced IL‐12p40 production, both gC1q and cC1q, and/or some specialized conformation of native C1q may be required for the inhibition. While C1q did not affect mRNA expression of Toll‐like receptor 4 (TLR4), MD‐2, and myeloid differentiation factor 88 (MyD88), BMDC treated with C1q showed the reduced activity of NF‐κB and the delayed phosphorylation of p38, c‐Jun N‐terminal kinase, and extracellular signal‐regulated kinase after LPS‐stimulation. CpG oligodeoxynucleotide‐induced IL‐12p40 and TNF‐α production, another MyD88‐dependent TLR‐mediated signal, was also suppressed by C1q treatment. Therefore, C1q is likely to suppress MyD88‐dependent pathway in TLR‐mediated signals. In contrast, C1q failed to suppress colony formation of B cells responding to LPS or LPS‐induced CD40 and CD86 expression on BMDC in MyD88‐deficient mice, indicating that inhibitory effects of C1q on MyD88‐independent pathways may be limited. Taken together, C1q may regulate innate and adaptive immune systems via modification of signalsmediated by interactions between invading pathogens and TLR.

Antiviral activity of limitin against encephalomyocarditis virus, herpes simplex virus, and mouse hepatitis virus: diverse requirements by limitin and alpha interferon for interferon regulatory factor 1.

ABSTRACT Limitin has sequence homology with alpha interferon (IFN-α) and IFN-β and utilizes the IFN-α/β receptor. However, it has no influence on the proliferation of normal myeloid and erythroid progenitors. In this study, we show that limitin has antiviral activity in vitro as well as in vivo. Limitin inhibited not only cytopathic effects in encephalomyocarditis virus- or herpes simplex virus (HSV) type 1-infected L929 cells, but also plaque formation in mouse hepatitis virus (MHV) type 2-infected DBT cells. In addition, administration of limitin to mice suppressed MHV-induced hepatitis and HSV-induced death. The antiviral activity may be mediated in part by 2′,5′-oligoadenylate synthetase, RNA-dependent protein kinase, and Mx protein, which inhibit viral replication or degrade viral components, because limitin induced their mRNA expression and enzyme activity. While limitin has antiviral activity as strong as that of IFN-α in vitro (the concentration that provided 50% inhibition of cytopathic effect is ∼30 pg/ml), IFN regulatory factor 1 (IRF-1) dependencies for induction of an antiviral state were different for limitin and IFN-α. In IRF-1-deficient fibroblasts, a higher concentration of limitin than of IFN-α was required for the induction of antiviral activity and the transcription of proteins from IFN-stimulated response element. The unique signals and the fewer properties of myelosuppression suggest that a human homolog of limitin may be used as a new antiviral drug.

Antifungal Activity of Micafungin in Serum

ABSTRACT We have evaluated the antifungal activity of micafungin in serum by using the disk diffusion method with serum-free and serum-added micafungin standard curves. Serum samples from micafungin-treated patients have been shown to exhibit adequate antifungal activity, which was in proportion to both the applied dose and the actual concentration of micafungin measured by high-performance liquid chromatography. The antifungal activity of micafungin in serum was also confirmed with the broth microdilution method.

Identification of amino-terminal region of adiponectin as a physiologically functional domain

Adiponectin is an abundant adipose‐specific protein, which acts as an anti‐diabetic, anti‐atherogenic, and anti‐inflammatory adipokine. Although recent advances in the field of adiponectin have been made by the identification of adiponectin receptors and by the understanding about relationship between its multimerization and functions, detailed molecular background remains unclear. Our established anti‐human adiponectin antibodies, ANOC 9103 and ANOC 9104, blocked some adiponectin functions such as the growth inhibition of B‐lymphocytes on stromal cells and the inhibition of acetylated LDL uptake in macrophages, suggesting that they may recognize important functional regions of adiponectin. As a result of epitope mapping based on the ability to bind to the deleted adiponectin mutants, we identified that these antibodies recognize amino‐terminal region of adiponectin before the beginning of the collagen‐like domain. Notably, a peptide fragment (DQETTTQGPGVLLPLPKGACTGWMA) corresponding to amino acid residues 17–41 of human adiponectin could bind to restricted types of cells and block adiponectin‐induced cyclooxygenase‐2 gene expression and prostaglandin E2 production in MS‐5 stromal cells. Moreover, the deletion of its amino‐terminal region reduced the abilities to inhibit not only collagen‐induced platelet aggregation but also diet‐induced hepatic steatosis. These data indicate that amino‐terminal region of adiponectin is a physiologically functional domain and that a novel receptor, which recognizes amino‐terminal region of adiponectin, may exist on some types of cells. Further investigations will contribute to the understanding of molecular mechanisms about adiponectin functions as well as to the designing of novel strategies for the treatment of patients with insulin‐resistance, vascular dysfunction, and chronic inflammation. J. Cell. Biochem. 98: 194–207, 2006.

Localized relapse in bone marrow in a posttransplantation patient with t(6;9) acute myeloid leukemia

We report a 38-year-old woman with t(6;9) acute myeloid leukemia who relapsed with localized leukemic cell growth in the bone marrow after she had undergone allogeneic bone marrow transplantation. The localized cell growth was first recognized by an apparent discrepancy in the DEK-CAN fusion transcript levels between the aspirates from the left and right iliac bone marrow. Magnetic resonance imaging of the iliac bone revealed localized cell accumulation in the left side. The nonhomogeneous and localized leukemic cell growth in this case may have been due to the graft-versus-leukemia effect following allogeneic transplantation with donor lymphocyte infusion. Serial monitoring of molecular markers for leukemia at different sites or magnetic resonance imaging of the bone marrow may be of value in detecting this type of relapse.

Immune-mediated peripheral neuropathy occurring simultaneously with recurrent graft-versus-host disease after allogenic hematopoietic stem cell transplantation

Peripheral neuropathy is a rare but important complication f allogenic hematopoietic stem cell transplantation (allo-HSCT). lthough it is potentially curable, it sometimes has a tremendous mpact on a patient’s quality of life. There have been reports of nflammatory peripheral neuropathy associated with chronic graftersus-host disease (GVHD), but its pathogenesis has not been ully revealed. We report a case of immune-mediated neuropahy after allo-HSCT for Philadelphia-chromosome-positive acute ymphoblastic leukemia (Ph-ALL). After conditioning with cyclophosphamide + total body irradition, a 44-year-old woman in the second molecular complete emission (mCR) of Ph-ALL underwent allogenic peripheral blood ematopoietic stem cell transplantation from her sister, with a oneocus mismatch of histocompatibility leukocyte antigen-B antigen. yclosporine A (CsA), short-term methotrexate, and mycophenoatemofetil were administered for GVHD prophylaxis. Grade 3 acute VHD (skin: stage 1, liver: stage 0, gut: stage 2) had diagnosed on ay24 had occurred and was treated effectively with methylpredisolone (mPSL) 2 mg/kg/day. Thereafter, no recurrence of GVHD ymptoms had been seen, despite a gradual reduction in the doses f CsA and steroid, and bone marrow examinations had revealed hat the patient continued to be in mCR from Ph-ALL (Fig. 1). On day 240, the patient developed a self-limiting common cold. rom around day 250, she became aware of gradually worsening uscle weakness in the left upper extremity (at the time CsA was 0 mg b.i.d. and steroid was 10 mg q.d. as prednisolone, CsA had still ept on being reduced every two weeks). At a regular consultation n day 273, GVHD symptoms (generalized skin rash, dry eyes, and tomatitis) were noticed. Blood examination showed extremely levated hepatic enzymes (AST 192 U/L [normal range: 13–33 U/L], LT 408 U/L [6–30 U/L], -GTP 766I U/L [10–47 U/L], ALP 1018 U/L 115–359 U/L], and T-bil 0.6 mg/L [0.2–1.0 mg/L]). As for immune econstitution, serum IgG was 955 mg/dL [870–1700 mg/dL], WBC as 7700/ L [3800–8500/ L] and lymphocyte was 14% [20–45%]. ymphocyte subsets were not examined. The patient was admited to hospital and underwent skin biopsy and echo-guided liver iopsy. The biopsy specimens were consistent with GVHD. Manual uscle testing (MMT) showed a weakness of 4 in the left trapezus, 3 in the left deltoid muscle, 0 in the left biceps and 4 in the

An indolent subtype of “intravascular lymphoma”: A case with a 3-year history of LDH elevation

A 59-year-old woman, without any past history of serious illnesses, was admitted for examination of high serum level of lactate dehydrogenase (LDH). She had undergone regular annual medical checkup for recent 10 years, and no abnormality had been pointed out until high LDH (472 IU/l; normal range: 214 – 370 IU/l) was detected in August 2003. Her LDH level further elevated to 543 IU/l in July 2004 and 871 IU/l (normal range: 101 – 236 IU/l) in August 2005, while any other abnormality was not detected [Figure 1(a)]. She consulted our hospital in November 2005 in regard to high LDH. While she was followed up as an outpatient, she occasionally complained of a subfever and general fatigue. Her LDH level gradually rose to 1355 IU/l (LDH subtypes 2 and 3 were dominant) in February 2006, and the value of soluble interleukin-2 receptor (sIL2-R) was 1870 U/ml. Since a malignant tumor, such as lymphoma, was the most possible cause for the presence of high LDH and sIL2-R, we had thoroughly investigated the existence of malignancy in her body. Gastroduodenoscopy and colonoscopy revealed no significant abnormality. Both of fluorodeoxyglucose positron emission tomography (FDG-PET) and gallium scintigraphy showed no abnormal hot spot. A computed tomographic scan (CT) of the chest and the abdomen showed the presence of 15-mm mass lesion in the uterus, which was compatible with uterine myoma, and no abnormality in the lung, mediastinum, liver, pancreas, kidney, and spleen. Bone marrow aspiration and biopsy showed neither invasion of malignant cell nor hemophagocytosis. She was admitted to our hospital for further examination in April 2006. On admission, there was no surface lymphadenopathy or skin lesion. Complete blood count was within normal limit, and its morphology was normal. LDH was 1412 IU/l (subtypes 1:2:3:4:5 were 18.5%: 35.0%:32.0%:12.2%:2.4%, respectively), and liver transaminase and C-reactive protein were slightly elevated. Principal tumor markers such as CEA, CA19-9, and AFP were normal except for neuron specific enolase (NSE: 42.3 ng/ml; normal range 510). Cytology of the uterus revealed abnormal lymphocytes, which suggested the presence of lymphoma. However, these abnormal lymphocytes were never detected again with several sequential cytology of the uterus, and even with the dilatation and curettage biopsy. Since serum LDH level varied from 1000 to 1500 IU/l without increasing tendency during this admission, she was discharged in May 2006 and followed up for the time-being. During two months of follow-up, her LDH gradually increased again, and she came to have subfever consistently. In July 2006, she began to feel difficulty in speech and to be forgetful; therefore she was admitted again for further examination. Brain magnetic resonance imaging (MRI) showed normal appearance [Figure 1(b)]. Cerebrospinal fluid (CSF) examination revealed slight elevation of protein level, but CSF cytopathology was negative for malignant cells. Splenomegaly was found on abdominal CT, and liver transaminases were also elevated on blood chemistry examination. Since we suspected splenic

Successful treatment with rituximab for angioimmunoblastic T-cell lymphoma

We experienced a patient with angioimmunoblastic T-cell lymphoma (AITL) without Epstein-Barr virus-positive B (EBV-B) cells at initial presentation who progressed to AITL with expansion of EBV-B cells at relapse. Based on the results of repeated biopsy, the patient was successfully treated with rituximab in combination with chemotherapy at relapse. A repeat biopsy may be necessary to determine the optimum therapeutic strategy at relapse, particularly for patients with suspected expansion of B cell and/or EBV-B cells. Although a recent report found no significant prognostic advantage of rituximab, it is one of the active drugs for selected patients with AITL.

Reversible posterior leukoencephalopathy syndrome of bilateral thalamus in acute lymphoblastic leukemia

Numerous cases of reversible posterior leukoencephalopathy syndrome (RPLS) are reported in patients having risk factors such as malignant hypertension, eclampsia, receiving solid organ/hematopoietic stem cell transplant, and exposure to chemotherapeutic agents and immunosuppressive drugs. We experienced a very interesting case of atypical RPLS in the thalamus bilaterally accompanied by brain hemorrhage in acute lymphoblastic leukemia (ALL), and described here. A 62-year-old Japanese female was referred to our hospital with fever, general fatigue and leukocytosis in May 2011. Her white blood cell count was 139.6 10 9 /L, of which lymphoblasts constituted 96%, hemoglobin level was 8.2 g/ dL and platelet count was 15.0 10 9 /L. Leukemic lymphoblasts were peroxidase negative and their surface markers were positive for CD10, CD19, CD34 and human leukocyte antigen (HLA)-DR. Th e karyotype was 46, XY; the BCR/ABL fusion gene was not detected. Th e patient was diagnosed as having ALL and treated with cyclophosphamide, vincristine, adriamycin and prednisolone. Although leukemia cells were not present in peripheral blood at 9 days after starting chemotherapy, febrile neutropenia occurred, and antibiotic treatment was started immediately. After the administration of antibiotic and antifungal agents, erythroderma developed on her trunk, extremities and back. Stevens – Johnson syndrome (SJS) was diagnosed, and high-dose methylprednisolone was given. Hyponatremia (124 mEq/L) also occurred at this time, as an eff ect of the chemotherapeutic agents. Although the erythroderma and hyponatremia improved slowly over the next 4 days, her level of consciousness decreased gradually before falling suddenly, accompanied by high fever. Blood culture was positive for Pseudomonas aeruginosa while cerebrospinal fl uid (CSF) culture was negative. Th e patient ’ s level of consciousness improved after antibiotic treatment for sepsis; however, she lost consciousness once more, accompanied by high blood pressure (180/118 mmHg). Emergency computed tomography (CT) of the brain revealed multiple low-density areas in the thalamus, midbrain and pons bilaterally. Magnetic resonance imaging (MRI) of the brain performed 2 days after the CT scan showed multiple high signal intensity lesions with small hemorrhagic areas on T2-weighted fl uid attenuated inversion recovery imaging, T2 star-weighted imaging and diff usion-weighted imaging (Figure 1). No thrombosis of the bilateral internal vein and Galen vein was detected in brain CT and MRI. At 10 days after completion of treatment for sepsis and control of blood pressure, the patient was fully conscious. She was diagnosed with hemorrhage in atypical RPLS. MRI obtained 2 months after treatment showed no RPLS lesions