Daphnetin inhibits invasion and migration of LM8 murine osteosarcoma cells by decreasing RhoA and Cdc42 expression.

Daphnetin, 7,8-dihydroxycoumarin, present in main constituents of Daphne odora var. marginatai, has multiple pharmacological activities including anti-proliferative effects in cancer cells. In this study, using a Transwell system, we showed that daphnetin inhibited invasion and migration of highly metastatic murine osteosarcoma LM8 cells in a dose-dependent manner. Following treatment by daphnetin, cells that penetrated the Transwell membrane were rounder than non-treated cells. Immunofluorescence analysis revealed that daphnetin decreased the numbers of intracellular stress fibers and filopodia. Moreover, daphnetin treatment dramatically decreased the expression levels of RhoA and Cdc42. In summary, the dihydroxycoumarin derivative daphnetin inhibits the invasion and migration of LM8 cells, and therefore represents a promising agent for use against metastatic cancer.

Inhibition of monocarboxylate transporter 1 suppresses the proliferation of glioblastoma stem cells.

Recent evidence suggests that a minor subset of cancer cells, termed cancer stem cells (CSCs), have self-renewal and tumorigenic potential. Therefore, the characterization of CSCs is important for developing therapeutic strategies against cancer. Cancer cells rely on anaerobic glycolysis to produce ATP even under normoxic conditions, resulting in the generation of excess acidic substances. Cancer cells maintain a weakly alkaline intracellular pH to support functions. Glioblastoma is an aggressive malignancy with a poor 5-year survival rate. Based on the hypothesis that ion transport-related molecules regulate the viability and function of CSCs, we investigated the expression of ion transport-related molecules in glioblastoma CSCs (GSCs). Quantitative RT-PCR analysis showed that monocarboxylate transporter1 (MCT1) were upregulated in GSCs, and inhibition of MCT1 decreased the viability of GSCs compared with that of non-GSCs. Our findings indicate that MCT1 is involved in the maintenance of GSCs and is a promising therapeutic target for glioblastoma.

Effect of minodronate on the speed of sound of the calcaneus in postmenopausal women with an increased risk of fractures: A clinical practice-based observational study.

BACKGROUND: We previously reported that alendronate and risedronate reduce the urinary levels of cross-linked N-terminal telopeptides of type I collagen (NTX) by 44.9% and 34.7%, respectively, at 3 months after the start of treatment, and increase the speed of sound (SOS) of the calcaneus by 0.6% and 0.65%, respectively, at 12 months after the start of treatment in postmenopausal women with osteoporosis. The aim of the present clinical practice-based observational study was to examine the effect of treatment with minodronate for 12 months on the SOS of the calcaneus and on bone turnover markers in postmenopausal women with an increased risk of fractures. METHODS: Forty-two postmenopausal women with osteoporosis or osteopenia with a clinical risk factor for fractures who had been treated with minodronate for > 12 months were enrolled in the study. The SOS and bone turnover markers were monitored during treatment with minodronate for 12 months. RESULTS: Compared to their baseline values, the urinary levels of NTX at 3 months and the serum levels of alkaline phosphatase at 12 months were significantly decreased at 47.5% and 25.8%, respectively. At 12 months, the SOS increased modestly, but significantly, by 0.47%, compared to the baseline value. CONCLUSION: The present study confirmed that minodronate suppressed bone turnover and modestly increased the SOS of the calcaneus in postmenopausal women with an increased risk of fractures.

Targeting the canonical Wnt/ß-catenin pathway in hematological malignancies.

The canonical Wnt/ß-catenin pathway plays an important role in different developmental processes through the regulation of stem cell functions. In the activation of the canonical Wnt/ß-catenin pathway, ß-catenin protein is imported into the nucleus and activates transcription of target genes including cyclin D1 and c-myc. Aberrant activation of the Wnt/ß-catenin pathway contributes to carcinogenesis and malignant behaviors, and Wnt signaling is essential for the maintenance of cancer stem cells. The canonical Wnt/ß-catenin pathway has been investigated extensively as a target in cancer treatment and several specific inhibitors of this signaling pathway have been identified through high-throughput screening. In this review, the significance of the canonical Wnt/ß-catenin pathway in hematological carcinogenesis and screening methods for specific inhibitors are discussed.

Vitamin K nutritional status and undercarboxylated osteocalcin in postmenopausal osteoporotic women treated with bisphosphonates.

Serum undercarboxylated osteocalcin (ucOC) is an index of vitamin K nutritional status in treatment-naive postmenopausal osteoporotic women. The purpose of the present study was to reveal the association between vitamin K nutritional status and serum ucOC concentrations in postmenopausal osteoporotic women taking bisphosphonates. Eighty-six postmenopausal women with osteoporosis (age range: 47-90 years) initiated bisphosphonate treatment. Vitamin K nutritional status was evaluated using a simple vitamin K-intake questionnaire and serum ucOC concentrations were measured after 6 months of treatment. The patients were divided into two groups according to the simple vitamin K-intake questionnaire score: a low vitamin K-intake (score <40) group (n=67) and a normal vitamin K-intake (score >=40) group (n=19). There were no significant differences between the groups in baseline parameters including age, height, body weight, body mass index, serum alkaline phosphatase (ALP), urinary cross-linked N-terminal telopeptides of type I collagen (NTX), and changes in serum ALP and urinary NTX concentrations during the 6-month treatment period. However, the mean serum ucOC concentration after 6 months of treatment was significantly higher in the low vitamin K-intake group (2.79 ng/mL) than in the normal vitamin K-intake group (2.20 ng/mL). These results suggest that 78% of postmenopausal osteoporotic women treated with bisphosphonates may have vitamin K deficiency as indicated by low vitamin K-intake and high serum ucOC concentrations, despite having a similar reduction in bone turnover to women who have normal vitamin K-intake.

Effect of risedronate on speed of sound in postmenopausal women with osteoporosis.

AIM: To examine the effects of treatment with risedronate for 1 year on speed of sound (SOS) of the calcaneus and bone turnover markers in postmenopausal women with osteoporosis. METHODS: Thirty-eight postmenopausal women with osteoporosis who had been treated with risedronate for > 1 year were enrolled in the study. The SOS and bone turnover markers were monitored during treatment with risedronate for 1 year. RESULTS: The urinary levels of cross-linked N-terminal telopeptides of type I collagen and serum levels of alkaline phosphatase were significantly decreased at 3 mo (-34.7%) and 12 mo (-21.2%), respectively, compared with the baseline values. The SOS increased modestly, but significantly by 0.65% at 12 mo compared with the baseline value. Treatment with risedronate elicited an increase in the SOS of the calcaneus exceeding the coefficient of variation in vivo (0.27%). CONCLUSION: The present study confirmed that risedronate suppressed bone turnover and elicited a clinically significant increase in the SOS of the calcaneus in postmenopausal women with osteoporosis.

[Current state and the future of medical technologist as a specialist Japanese Association of Medical Technologists].

The recognition mechanism is composed of seven groups that conduct qualifying examinations within each region. The average pass rate in three qualifying examination areas conducted by JAMT is 77.2%. It is necessary to integrate similar qualifying examinations in the future and the new "Integrated management technologist system" has a key role from the aspect of personnel training. Requirements for the integrated management inspection technologist are as follows: 1) Person who obtains many positive evaluations; 2) Excellent personality; 3) Person with a sense of justice; 4) Person with resolution-making abilities and decision; 5) Person who can see the heart of an issue; 6) Person who has the potential to become a leader; 7) Person with crisis-management ability. Also, selected personnel are expected to become leaders not only in their field of expertise but also within their hospital and JAMT management.

Influence of treatment with alendronate on the speed of sound, an ultrasound parameter, of the calcaneus in postmenopausal Japanese women with osteoporosis: a clinical practice-based observational study.

PURPOSE: The influence of alendronate (ALN) treatment on the quantitative ultrasound parameters of the calcaneus remains to be established in Japanese patients. The aim of the present clinical practice-based observational study was to examine the influence of ALN treatment for 1 year on the speed of sound (SOS) of the calcaneus and bone turnover markers in postmenopausal Japanese women with osteoporosis. PATIENTS AND METHODS: Forty-five postmenopausal Japanese women with osteoporosis who had received treatment with ALN for more than 1 year were enrolled in the study. The SOS and bone turnover markers were monitored over 1 year of ALN treatment. RESULTS: The urinary levels of cross-linked N-terminal telopeptides of type I collagen and serum levels of alkaline phosphatase decreased significantly from the baseline values (-44.9% at 3 months and -22.2% at 12 months, respectively). The SOS increased modestly, but significantly, from the baseline value (0.6% at both 6 and 12 months). The percentage decrease in the urinary levels of cross-linked N-terminal telopeptides of type I collagen at 3 months was significantly correlated with the percentage increase in the SOS only at 6 months (correlation coefficient, 0.299). CONCLUSION: The present study confirmed that ALN treatment suppressed bone turnover, producing a clinically significant increase in the SOS of the calcaneus in postmenopausal Japanese women with osteoporosis.

Microglial Amyloid-ß1-40 Phagocytosis Dysfunction Is Caused by High-Mobility Group Box Protein-1: Implications for the Pathological Progression of Alzheimer's Disease.

In Alzheimer disease (AD) patient brains, the accumulation of amyloid-ß (Aß) peptides is associated with activated microglia. Aß is derived from the amyloid precursor protein; two major forms of Aß, that is, Aß1-40 (Aß40) and Aß1-42 (Aß42), exist. We previously reported that rat microglia phagocytose Aß42, and high mobility group box protein 1 (HMGB1), a chromosomal protein, inhibits phagocytosis. In the present study, we investigated the effects of exogenous HMGB1 on rat microglial Aß40 phagocytosis. In the presence of exogenous HMGB1, Aß40 markedly increased in microglial cytoplasm, and the reduction of extracellular Aß40 was inhibited. During this period, HMGB1 was colocalized with Aß40 in the cytoplasm. Furthermore, exogenous HMGB1 inhibited the degradation of Aß40 induced by the rat microglial cytosolic fraction. Thus, extracellular HMGB1 may internalize with Aß40 in the microglial cytoplasm and inhibit Aß40 degradation by microglia. This may subsequently delay Aß40 clearance. We further confirmed that in AD brains, the parts of senile plaques surrounded by activated microglia are composed of Aß40, and extracellular HMGB1 is deposited on these plaques. Taken together, microglial Aß phagocytosis dysfunction may be caused by HMGB1 that accumulates extracellularly on Aß plaques, and it may be critically involved in the pathological progression of AD.

Clinical and histopathological features of myopathies in Japanese patients with anti-SRP autoantibodies.

To elucidate the clinical and histopathological features associated with autoantibodies to the signal recognition particle (SRP), we have studied 23 Japanese patients with this specificity among 3,500 patients with polymyositis/dermatomyositis and other connective tissue diseases. Anti-SRP antibodies were determined based on analysis of RNA and protein components by immunoprecipitation assays. The pathological analysis was performed by using special stainings including alkaline phosphatase, myosin ATPase, and modified Gomori trichrome stainings. Twenty-one (92%) of these 23 patients had myositis, 8 of whom (38%) required cytotoxic agents or intravenous immunoglobulin therapy in addition to corticosteroid therapy. Four patients (16%) had rheumatoid arthritis, two of whom had no features of myositis. Muscle biopsy specimens of 11 patients were examined histologically in detail. All 11 had muscle fiber necrosis and/or regeneration, but only one had infiltration of inflammatory cells. Six of the 11 (55%) patients showed type I fiber predominance by ATPase staining, while eight control myositis patients without anti-SRP antibodies did not. There was no correlation of other neurogenic features in histology with the presence of anti-SRP antibodies. These studies suggest that anti-SRP autoantibodies are most likely to be related to myopathies that are resistant to corticosteroid therapy and without inflammation histopathologically.

Longterm effect of intermittent cyclical etidronate therapy on corticosteroid-induced osteoporosis in Japanese patients with connective tissue disease: 7-year followup.

OBJECTIVE: To determine the efficacy and safety of intermittent cyclical etidronate therapy of up to 7 years for corticosteroid-induced osteoporosis. METHODS: One hundred two Japanese patients who originally participated in a 3-year prospective randomized study were enrolled into an open-label followup study. All patients had received > 7.5 mg of prednisolone daily for at least 90 days before entry into the original study and were randomly assigned to 2 treatment arms: E, those receiving etidronate disodium (200 mg per day) for 2 weeks together with 3.0 g of calcium lactate and 0.75 microg of alphacalcidol daily; and C, controls receiving only the latter. Endpoints included changes from baseline in bone mineral density (BMD) of the lumbar spine and the rate of new vertebral fractures. RESULTS: The mean (+/- SD) lumbar spine BMD had increased by 5.9% +/- 8.8% (p = 0.00007) and 2.2% +/- 5.8% (p = 0.013) from baseline after 7 years in groups E and C, respectively. This improvement in BMD in group E was significantly better than in group C (p = 0.02). The frequency of new vertebral fractures was lower in group E, resulting in reduction of the risk of such new fractures by 67% at year 7 (odds ratio 3.000; 95% confidence interval, 0.604 14.90; p = 0.18). There were no severe adverse events in group E during our study. CONCLUSION: Our results indicate that longterm (up to 7 years) intermittent cyclical etidronate therapy is safe and effective for prevention and treatment of corticosteroid-induced osteoporosis in patients with connective tissue diseases.

Clinical and immunogenetic features of patients with autoantibodies to asparaginyl-transfer RNA synthetase.

OBJECTIVE: We have previously described anti-KS autoantibodies and provided evidence that they are directed against asparaginyl-transfer RNA (tRNA) synthetase (AsnRS). The aim of the present study was to identify patients with anti-AsnRS autoantibodies and elucidate the clinical significance of this sixth antisynthetase antibody. In particular, we studied whether it was associated with the syndrome of myositis (polymyositis or dermatomyositis [DM]), interstitial lung disease (ILD), arthritis, and other features that had been previously associated with the 5 other anti-aminoacyl-tRNA synthetase autoantibodies. METHODS: More than 2,500 sera from patients with connective tissue disease (including myositis and ILD) and controls were examined for anti-AsnRS autoantibodies by immunoprecipitation (IP). Positive and control sera were tested for the ability to inhibit AsnRS by preincubation of the enzyme source with the serum. The HLA class II (DRB1, DQA1, DQB1, DPB1) alleles were identified from restriction fragment length polymorphism of polymerase chain reaction-amplified genomic DNA. RESULTS: Anti-AsnRS antibodies were identified in the sera of 8 patients (5 Japanese, 1 American, 1 German, and 1 Korean) by IP of the same distinctive set of tRNA and protein that differed from those precipitated by the other 5 antisynthetases, and these antibodies showed specific inhibition of AsnRS activity. Two of these patients had DM, but 7 of 8 (88%) had ILD. Four patients (50%) had arthritis, and 1 had Raynaud's phenomenon. This antisynthetase was very rare among myositis patients (present in 0% of Japanese myositis patients), but it was found in 3% of Japanese ILD patients. Thus, most patients with anti-AsnRS had chronic ILD with or without features of connective tissue disease. Interestingly, all 4 Japanese patients tested had DR2 (DRB1*1501/1502), compared with 33% of healthy controls. CONCLUSION: These results indicate that anti-AsnRS autoantibodies, like anti-alanyl-tRNA synthetase autoantibodies, have a stronger association with ILD than with myositis and may be associated with the DR2 phenotype.