The production of coagulation factor VII by adipocytes is enhanced by tumor necrosis factor-α or isoproterenol.

BACKGROUND: A relationship has been reported between blood concentrations of coagulation factor VII (FVII) and obesity. In addition to its role in coagulation, FVII has been shown to inhibit insulin signals in adipocytes. However, the production of FVII by adipocytes remains unclear. OBJECTIVE: We herein investigated the production and secretion of FVII by adipocytes, especially in relation to obesity-related conditions including adipose inflammation and sympathetic nerve activation. METHODS: C57Bl/6J mice were fed a low- or high-fat diet and the expression of FVII messenger RNA (mRNA) was then examined in adipose tissue. 3T3-L1 cells were used as an adipocyte model for in vitro experiments in which these cells were treated with tumor necrosis factor-α (TNF-α) or isoproterenol. The expression and secretion of FVII were assessed by quantitative real-time PCR, Western blotting and enzyme-linked immunosorbent assays. RESULTS: The expression of FVII mRNA in the adipose tissue of mice fed with high-fat diet was significantly higher than that in mice fed with low-fat diet. Expression of the FVII gene and protein was induced during adipogenesis and maintained in mature adipocytes. The expression and secretion of FVII mRNA were increased in the culture medium of 3T3-L1 adipocytes treated with TNF-α, and these effects were blocked when these cells were exposed to inhibitors of mitogen-activated kinases or NF-κB activation. The ß-adrenoceptor agonist isoproterenol stimulated the secretion of FVII from mature adipocytes via the cyclic AMP/protein kinase A pathway. Blockade of secreted FVII with the anti-FVII antibody did not affect the phosphorylation of Akt in the isoproterenol-stimulated adipocytes. CONCLUSION: Obese adipose tissue produced FVII. The production and secretion of FVII by adipocytes was enhanced by TNF-α or isoproterenol via different mechanisms. These results indicate that FVII is an adipokine that plays an important role in the pathogenesis of obesity.

APTT reagent with ellagic acid as activator shows adequate lupus anticoagulant sensitivity in comparison to silica-based reagent.

BACKGROUND: Lupus anticoagulant (LA) is an antibody that interferes with phospholipid-dependent coagulation reactions. Activated partial thromboplastin time (APTT) is widely used as a test for LA screening. APTT reagents are composed of activators, such as silica or ellagic acid, and phospholipids, and APTT reagents with silica are recommended for LA screening because of greater sensitivity. However, the effects of activators on LA activity have not been adequately investigated. OBJECTIVES: In this study, we examined whether an ellagic acid-based reagent was highly sensitive to LA in a low phospholipid condition and useful for LA screening. METHODS: Silica-based (SL) and ellagic acid-based (EA) reagents were prepared in-house with the same composition and concentration of phospholipids, while the commercial APTT reagents APTT-SLA (SLA), Actin FSL (FSL), APTT-SP (SP) and PTT-LA (PTT) were also included in the study. RESULTS: The normal reference ranges for SL and EA were 30.1-47.0 and 28.0-40.2 s, respectively, while the cut-off index values for circulating anticoagulant activity (ICA) calculated from the results obtained with SL, EA, SLA, FSL, SP and PTT were 12.9, 11.5, 13.2, 15.6, 14.3 and 14.0, respectively. The sensitivity of those reagents based on those cut-off values was 91%, 96%, 68%, 46%, 91% and 86%, respectively. CONCLUSIONS: Our results showed that the ellagic acid-based reagent was more sensitive to LA than silica-based reagents in a low phospholipid condition and had adequate sensitivity to detect LA. We concluded that the sensitivity of APTT reagents for LA is dependent on phospholipid concentration and not the activator.

Establishment and characterization of a new human clear-cell sarcoma cell-line, HS-MM.

We have established and morphologically characterized a new human clear-cell sarcoma cell-line, HS-MM, from the pleural effusion of a 39-year-old man with pulmonary metastasis derived from the primary popliteal tumour. The HS-MM cells were round or spindle-shaped, with round nuclei containing extremely prominent nucleoli. Light microscopically, the heterotransplanted nude mouse tumours showed essentially the same features as those of the original sarcoma, revealing an alveolar pattern of tumour cells with abundant clear cytoplasm. Both in vitro and in vivo, the cells reacted with anti-S-100 protein and melanoma-specific HMB 45 antibodies by immunohistochemistry. Ultrastructurally, the cells contained round euchromatin-rich nuclei with large nucleoli revealing conspicuous nucleolonema, and a large amount of glycogen and a few lysosomal dense bodies, but no premelanosomes in their cytoplasm. The HS-MM cell line was thus fully proven to exhibit the unique characteristics of a clear sarcoma both in vitro and in vivo, being also compatible with an amelanotic melanoma. This cell line will therefore be extremely useful for clinicopathological and histogenetic studies on clear cell sarcomas.

Establishment and characterization of a new human synovial sarcoma cell line, HS-SY-II.

BACKGROUND: The line of differentiation in synovial sarcoma still remains controversial. Thus far, only a few human synovial sarcoma cell lines have been described. However, their morphologic characteristics have not been fully established. EXPERIMENTAL DESIGN: We established a new synovial sarcoma cell line (HS-SY-II) from pleural effusion with lung metastasis in a typical example of the monophasic spindle cell type. The HS-SY-II cells, in vitro and in vivo, were examined by light microscopy, immunohistochemistry, electron microscopy, and cytogenetics. RESULTS: The HS-SY-II cells showed a hypertriploid karyotype with complex chromosome abnormalities including pathognomonic t(X;18)(p11;q11), and have been stably maintained for more than 40 months in vitro, showing rather small spindle or polygonal shape without conspicuous pleomorphism. Histologic features of initially and serially transplanted tumors in nude mice were essentially the same as those of the original sarcoma, corresponding to the monophasic spindle cell variant with a prominent palisading pattern and calcified foci in parts. The HS-SY-II cells in vitro and in vivo similarly expressed vimentin and cytokeratin by immunohistochemistry, and also exhibit the same ultrastructural features such as irregularly shaped nuclei with prominent nucleoli, many paranuclearly running intermediate filaments, and filopodia-like processes. CONCLUSIONS: This HS-SY-II cell line retaining the distinct morphological characteristics as the monophasic spindle cell type of synovial sarcoma therefore will be extremely useful for various pathomorphologic investigations on synovial sarcoma.

[Concentrations of cefotiam and cefmenoxime in the human bone marrow blood and osseous tissue].

Prophylactic antimicrobial therapy for the prevention of infection during total joint arthroplasty or spinal surgery was investigated using cefotiam (CTM) and cefmenoxime (CMX), which are cephems having broad antimicrobial spectrum. Seven patients received 2 g of CTM, and 12 patients 2 g of CMX, by intravenous drip infusion over 15 minutes. Blood, bone marrow and osseous tissue samples were obtained at the end of the infusion, and 30, 60, 90 and 120 minutes after the beginning of the infusion. The concentrations of CTM and CMX were detected by the agar-well method. CTM and CMX were smoothly distributed from blood to bone marrow and osseous tissue, and kept on the effective concentrations for prophylaxis against postoperative infections. The primary wound healing was obtained in all cases, and no side effects were also recognized.

Nevoid basal cell epithelioma syndrome in Japan.

A 49-year-old Japanese man with typical features of nevoid basal cell epithelioma syndrome is reported. The patient had cystic lesions of the jaw bone, multiple basal cell epitheliomas of the scalp and face, pits on the palms and soles, bifid ribs, scoliosis and calcification of the falx cerebri. In Japan five certain cases have been reported previously. We added one new case and reviewed the other five cases briefly.