AIE phenomena of a cyanostilbene derivative as a probe of molecular assembly processes.

The initial processes of the crystallization of a solute molecule, 1-cyano-trans-1,2-bis-(4'-methylbiphenyl)-ethylene (CN-MBE) in binary solution (water and acetone), were investigated by means of fluorescence spectroscopy as well as scanning electron microscopy (SEM). With an increase in the volume fraction (Vw) of the poor solvent (water) in the solution, a drastic change in the fluorescence spectra and intensity of CN-MBE was observed. This change was attributed to aggregation induced emission (AIE). By analyzing the evolution of AIE by multivariate curve resolution-alternating least squares (MCR-ALS), it was revealed that four main species appeared in the solution depending on the Vw values. On the basis of molecular exciton theory, we assigned these four emissive states to the monomer, H-dimer, J-dimer, and H-aggregates. Interestingly, the J-dimer state was observed only in a Vw range of 40% to 50%, just before the formation of the aggregate. This result suggests that the J-dimer plays an important role as the precursor for larger aggregates leading to crystal formation. By integrating the present results with previous work on the crystallization of CN-MBA through solvent evaporation, we discussed the dynamics of the crystallization from the viewpoint of the sequence of molecular species appearing in the aggregation in solution.

Direct Visualization of the Two-step Nucleation Model by Fluorescence Color Changes during Evaporative Crystallization from Solution.

The two-step nucleation model for crystal nuclei formation explains several experimental and theoretical results better than the classical nucleation theory. We report here direct visualization of the two-step nucleation model for organic molecular crystallization. Evaporative crystallization from a solution of a dibenzoylmethane boron complex that displays mechanofluorochromism, a fluorescence color change induced by mechanical perturbation, was probed by fluorescence change. The dependence of fluorescence change on dispersion concentration of the complex in a polymer matrix was also investigated. We detected transitional emission from the amorphous cluster state prior to crystallization. This is the first demonstration of the two-step nucleation model based on fluorescence color changes.

Solvent Viscosity Effect on Triplet-Triplet Pair in Triplet Fusion.

The effect of the solvent viscosity dependence of time-resolved magnetoluminescence (ML) on the delayed fluorescence of 9,10-diphenylanthracene (DPA) sensitized by platinum octaethylporphyrin has clarified the structure and dynamics of the triplet-triplet pair (TT), i.e., the transition state of triplet fusion. Phase inversion of the ML effect with time provides evidence for the recycle dynamics of the excited triplet state for DPA in triplet fusion. The electron spin-relaxation by random molecular rotation causes intersystem crossing among the different spin states of the triplet-triplet pair and allows the (3,5)TT to engage in triplet fusion. Therefore, slow-down of the molecular diffusion by an increase in the solvent viscosity can enhance the triplet fusion yield. However, the reduction of the ML effect observed in quite high viscosity solvents suggests that the substantially slow rotational motion decreases the triplet fusion yield due to steric factors in electron exchange from the triplet-triplet pair.

Prognostic value of SS18-SSX fusion type in synovial sarcoma; systematic review and meta-analysis.

SS18-SSX (formerly called SYT-SSX) fusion gene has been established clinically as a molecular diagnostic test for synovial sarcoma, but the prognostic value of the fusion gene variant for survival is controversial. The objective of this systematic review is to provide an up-to-date and unprecedented summary of the prognostic impact of SS18-SSX fusion type in synovial sarcoma. Studies evaluating SS18-SSX fusion type as a prognostic marker in synovial sarcoma were systematically searched for in MEDLINE, EMBASE, and Web of Science. Comparative analysis of the pooled hazard ratios (HR) between fusion types was carried out, in order to assess the likelihood of overall survival (OS), disease-specific survival (DSS), progression-free survival (PFS), and metastasis-free survival (MFS). A total of 10 studies comprising 902 patients with synovial sarcoma were considered for the meta-analysis. The pooled HR for eight eligible studies evaluating for OS or DSS was 1.28 (95% confidence interval: 0.81-2.00), suggesting no significant difference between SS18-SSX1 and SS18-SSX2 (P = 0.29). For seven studies which evaluated for PFS or MFS, the presence of SS18-SSX1 may indicate a lower survival probability than that of SS18-SSX2, although the effect did not reach a level of statistical significance (P = 0.09). There was no significant difference in OS or DSS between SS18-SSX1 and SS18-SSX2, but there were indications of SS18-SSX1 being an unfavorable prognostic factor of PFS or MFS. Further studies including cohorts with a longer follow-up period are needed.

Quantitative (201)thallium scintigraphy for prediction of histological response to neoadjuvant chemotherapy in osteosarcoma; systematic review and meta-analysis.

PURPOSE: The histological assessment of tumor necrosis of the excised lesion after neoadjuvant chemotherapy is the most important prognostic factor for patients with osteosarcoma, but more early prognostic factors are needed for the adjustment of adjuvant treatment regimen. The objective of this systematic review is to provide an up-to-date and unprecedented summary of the value of (201)thallium ((201)Tl) scintigraphy for the preoperative evaluation of the chemotherapy response of osteosarcoma. METHODS: Studies evaluating (201)Tl scintigraphy for the preoperative evaluation of the chemotherapy response of osteosarcoma were systematically searched for in MEDLINE, EMBASE, and Web of Science. Pooled sensitivity and specificity for each study were calculated into 2 × 2 contingency tables. The DerSimonian-Laird random-effects method was used for determining the pooled diagnostic odds ratio and the area under curve (AUC) of the summary receiver operating characteristic (SROC) curve. RESULTS: A total of six studies with 139 patients who fulfilled all of the inclusion criteria were considered for the meta-analysis. The pooled sensitivity and specificity were 0.93 (95% CI, 0.83-0.98) and 0.63 (95% CI, 0.52-0.74), respectively. A significant difference was found between the good and poor responders in the diagnostic odds ratio. The SROC curve showed that the AUC was 0.840, indicating excellent diagnostic accuracy. There was no statistically significant heterogeneity among the six studies. CONCLUSIONS: The alteration ratio derived from (201)Tl scintigraphy was useful for evaluating the histological response of patients to neoadjuvant chemotherapy in osteosarcoma.

Prognostic value of PAX3/7-FOXO1 fusion status in alveolar rhabdomyosarcoma: Systematic review and meta-analysis.

PURPOSE: The objective of this systematic review is to provide an unprecedented summary of the prognostic impact of PAX3/7-FOXO1 fusion status in alveolar rhabdomyosarcoma. METHODS: Studies evaluating PAX3/7-FOXO1 fusion gene or its variants as a prognostic marker were systematically searched and comparative meta-analysis of overall survival was carried out. RESULTS: A total of 7 studies comprising 993 patients with rhabdomyosarcoma were included. Three eligible studies showed no significant difference of survival between fusion positive and negative alveolar rhabdomyosarcoma. Four eligible studies showed that PAX3-FOXO1 fusion variant may indicate a lower survival probability than PAX7-FOXO1, although the effect did not reach a level of statistical significance (pooled hazard ratios, 1.66; 95% CI, 0.95-2.89; p=0.07). CONCLUSIONS: There was no significant difference in the overall survival between patients with the positive and negative fusion gene, but there were indications of PAX3-FOXO1 being an unfavorable prognostic factor.

Does expression of glucose transporter protein-1 relate to prognosis and angiogenesis in osteosarcoma?

BACKGROUND: The survival of patients who present with nonmetastatic extremity osteosarcoma has dramatically improved, but there are some patients who do not respond to chemotherapy. The ability to identify patients with a poorer prognosis might allow us to target different therapy for these patients. Glucose transporter protein-1 (Glut-1), one of the key factors in glucose metabolism, has been reported to be an independent prognostic factor in various tumors. However, little is known about the role of the Glut-1 pathway in osteosarcoma. QUESTIONS/PURPOSES: We asked (1) if Glut-1 expression is a prognostic marker for survival in patients with osteosarcoma, and (2) if there is a relationship between Glut-1 expression and tumor angiogenesis. PATIENTS AND METHODS: Thirty-seven patients with resectable high-grade osteosarcomas treated between 1982 and 2007 were reviewed retrospectively. Patients were excluded if representative biopsy material and followup data were not available. The expression of Glut-1 and the number of CD34-positive microvessels for angiogenic activity were measured immunohistochemically. The median followup was 6 years 6 months (range, 11-211 months). Survival analyses were evaluated using the Kaplan-Meier method and the Cox proportional hazards model. The association between Glut-1 expression and microvessel density was analyzed using Student's t-test and chi-square test. For 12 (32.4%) of 37 patients with osteosarcoma, the expression of Glut-1 was positive, with four patients (10.8%) showing strong expression of Glut-1 protein. RESULTS: The expression of Glut-1 correlated with a shorter disease-free survival period (relative risk, 20.13; 95% CI, 1.77-229.3; p=0.0016). The microvessel density mean value of positive Glut-1 expression (mean±SD, 26.5±19.4) was lower than that of negative expression (mean±SD, 46.4±35.3; Student's t-test, p=0.038). When more than 50 was defined as a high microvessel density, positive expression of Glut-1 was significantly associated with low microvessel density (chi-square test, p=0.049). CONCLUSIONS: These findings indicate that Glut-1 is a potential predictor of survival in patients with osteosarcoma and that glucose metabolism may be negatively associated with angiogenesis. If substantiated in larger numbers of patients, these findings might stimulate the development of novel treatments for patients with a poorer prognosis. LEVEL OF EVIDENCE: Level III, prognostic study. See the Instructions for Authors for a complete description of levels of evidence.

A novel assessment method of serum alkaline phosphatase for the diagnosis of osteosarcoma in children and adolescents.

BACKGROUND: A high serum alkaline phosphatase (ALP) level is valuable for the diagnosis of osteosarcoma in adults, but its use in teenagers is problematic because ALP levels are affected by age, gender, and pubertal stage. Because serum acid phosphatase (ACP) shows the same tetraphasic pattern as serum ALP in children and adolescents, we presumed that serum levels of ALP and ACP would have a strong correlation and that the ratio of ALP to ACP (ALP/ACP) would show little variation and would be useful for the diagnosis of osteosarcoma in teenagers. The purpose of this study was to evaluate the correlation between the serum levels of ALP and ACP and to investigate the validity of ALP/ACP in the differential diagnosis of osteosarcoma in children and adolescents. METHODS: We retrospectively examined 538 patients aged 1-18 years, including 24 with osteosarcomas, 8 with other malignant bone tumors, 56 with benign bone tumors, and with 450 non-tumor lesions (controls). We evaluated the serum levels of ALP and ACP, both obtained by preoperative examination. RESULTS: There were significant correlations between the serum ALP and ACP levels in the controls (r = 0.805 in males and r = 0.860 in females). The ratios of ALP to ACP in the controls showed little variation with age. In ROC curve analysis, to discriminate between the osteosarcomas and the controls, the cutoff levels of serum ALP and ALP/ACP were 956 (U/l) and 50.9 in males and 748 (U/l) and 43.3 in females, respectively. The sensitivity and specificity of serum ALP and ALP/ACP in the differential diagnosis of osteosarcoma were 50.0 and 89.0%, and 60.0 and 94.1%, respectively in males, and 61.5 and 72.7%, and 69.2 and 84.2%, respectively, in females. CONCLUSIONS: The results suggest that ALP/ACP is more useful than the serum ALP level in diagnosing osteosarcoma because it is little affected by the age.

Therapeutic effect and risk factors for complications of excision in 76 patients with schwannoma.

BACKGROUND: Benign schwannoma is the most common tumor of peripheral nerves. However, the clinical course of excision and risk factors associated with postoperative neurological deficits are not well known. We evaluated the incidence of preoperative symptoms, the incidence of postoperative neurological deficits, and the risk factors of neurological deficits. METHODS: We retrospectively reviewed data of 76 patients with schwannomas treated at our institution. We reviewed the clinical characteristics, and postoperative results, and determined the possible risk factors influencing the development of complications. RESULTS: Excision of schwannoma improved the Tinel-like signs in 47 of 51 patients and spontaneous pain in 14 of 15. Eleven of 17 patients with sensory deficits showed complete recovery, but six continued to show deficits with or without improvement. Motor deficits that were observed in four patients persisted in one. New neurological deficits developed in 21 patients and persisted until final follow-up in 8. Tinel-like signs was the risk factor of surgery-related neurological deficits (p = 0.009). CONCLUSIONS: New deficits developed predominantly in patients with preoperative Tinel-like signs. Attention should be given to patients with the factor.

Extraosseous extension caused by epidural hematoma in Gaucher disease mimicking malignant bone tumor.

Gaucher disease is an inherited autosomal-recessive disorder caused by the defective hydrolysis of glucocerebroside. The resultant hepatosplenomegaly, hematological changes, and orthopedic complications are the predominant symptoms. However, extraosseous manifestation of Gaucher disease, mimicking malignant bone tumor, is supposed to be rare. No reports of extraosseous manifestation of Gaucher disease caused by epidural hematoma were identified in the English literature. A 64-year-old man visited a nearby clinic for low back pain and was referred to our tumor clinic on suspicion of malignant bone tumor on sacral MRI. MRI revealed a demarcated solid lesion extending into the surrounding soft tissues on both sides of the sacral roots. During preoperative examination, he suffered from pathologic fracture in right mid-femur. We performed internal fixation with intramedullary nailing, simultaneously harvesting tissue specimens. Histopathological analysis showed aggregates of Gaucher cells in the right femur and hematoma in the sacrum. Epidural hematoma in Gaucher disease, usually attributed to thrombocytopenia, is a rare manifestation of skeletal complication, mimicking malignant processes.

Diversity of angiogenesis among malignant bone tumors.

Several studies have demonstrated that angiogenesis assessed by microvessel density (MVD) correlates with patient prognosis in various types of cancer, whereas data regarding the relevance of angiogenesis and prognosis in malignant bone tumors are scarce and controversial. The aim of this study was to examine MVD in representative malignant bone tumors, such as osteosarcoma, chondrosarcoma and Ewing's sarcoma, in order to clarify the role of angiogenesis in prognosis. A total of 69 patients with malignant bone tumors, including 44 osteosarcomas, 20 chondrosarcomas and 5 Ewing's sarcomas, were reviewed retrospectively and treated at our hospital between 1980 and 2007. Biopsy or pre-chemotherapy surgical specimens were immunohistochemically stained with anti-CD34 antibody. The MVD values of osteosarcomas and Ewing's sarcomas were significantly higher compared to chondrosarcoma. In osteosarcomas with high MVD, American Joint Committee on Cancer stage IIA, good histological response to chemotherapy was significantly correlated with better disease-free survival, while MVD was closely associated with age and chemotherapy response. In chondrosarcomas, the surgical margin (marginal and intralesional), MVD (high), tumor size (≥8) and histological grade (grades 2 and 3) significantly correlated with a shorter disease-free survival, while MVD was closely associated with age and histological grade. These findings showed that osteosarcomas and Ewing's sarcomas were hypervascular, compared to chondrosarcomas. In osteosarcomas, hypervascularity induced good chemotherapy response, leading to better prognosis, while in chondrosarcomas, high MVD was associated with histological grade and predicted poor prognosis.

Correlation between p38 mitogen-activated protein kinase and human telomerase reverse transcriptase in sarcomas.

BACKGROUND: One of the major components of telomerase is the human telomerase reverse transcriptase (hTERT) as the catalytic protein. hTERT mRNA expression are reported to be associated with prognosis and tumor progression in several sarcomas. However, there is no clear understanding of the mechanisms of hTERT in human sarcomas. Recent studies have suggested that signals transmitted through p38 mitogen-activated protein kinase (MAPK) can increase or decrease hTERT transcription in human cells. The purpose of this study was to analyse the correlation between p38 MAPK and hTERT in sarcoma samples. METHODS: We investigated 36 soft tissue malignant fibrous histiocytomas (MFH), 24 liposarcomas (LS) and 9 bone MFH samples for hTERT and p38 MAPK expression. Quantitative detection of hTERT and p38 MAPK was performed by RT-PCR. RESULTS: There was a significant positive correlation between the values of hTERT and p38 MAPK in all samples (r = 0.445, p = 0.0001), soft tissue MFH (r = 0.352, p = 0.0352), LS (r = 0.704, p = 0.0001) and bone MFH samples (r = 0.802, p = 0.0093). Patients who had a higher than average expression of p38 MAPK had a significantly worse prognosis than other patients (p = 0.0036). CONCLUSIONS: p38 MAPK may play a role in up-regulation of hTERT, and therefore, p38 MAPK may be a useful marker in the assessment of hTERT and patients' prognosis in sarcomas.

Antitumor effects of telomerase inhibitor TMPyP4 in osteosarcoma cell lines.

Telomere studies in carcinomas have been extensively reported for prognostic utility and effective methods for targeting telomerase therapy has been described, but efficacy of telomerase inhibitor remained unknown in sarcoma cells. In this study, we investigated the effects of telomerase inhibitor cationic porphyrin TMPyP4 on telomerase activity, telomere length, cell growth, and apoptosis in osteosarcoma cell lines. TMPyP4 significantly inhibited telomerase activity in telomerase positive HOS and Saos-2, but not in MG-63. TMPyP4 significantly induced telomere shortening, and inhibition of the cell growth in HOS and Saos-2 with over 17% apoptosis rates. In terms of MG-63, TMPyP4 did not induce inhibition of both telomerase activity and cell growth, although it induced significant telomere shortening. Telomere length after treatment was 5.60 kb in HOS, 4.00 kb in Saos-2, and 9.89 kb in MG-63. These results may suggest that both telomerase activity loss and sufficient telomere shortening are necessary to inhibit cell growth in telomerase positive osteosarcoma cells. TMPyP4 did not induced telomere shortening but significantly inhibited the growth with 22.6% apoptosis rate in telomerase negative with extremely longer telomere-U2OS, may indicating the antitumor effect of TMPyP4 may be related to DNA damage including telomere dysfunction through G-quadruplex stabilization, independent on telomere length.

Interferon-α/ß receptor as a prognostic marker in osteosarcoma.

BACKGROUND: A large-scale randomized trial of adjuvant interferon-α therapy for patients with osteosarcoma has been initiated as a joint protocol by the European and American Osteosarcoma Study Group. Because the expression of functional interferon-α/ß receptor is necessary for interferon-α agents to interact with osteosarcoma cells, we examined the expression of interferon-α/ß receptor in a series of osteosarcoma specimens. METHODS: Forty patients with high-grade resectable osteosarcoma, from whom surgical specimens had been obtained at the time of biopsy, were included in this retrospective study. Biopsy specimens were immunohistochemically stained with anti-interferon-α/ß receptor antibodies. Survival was estimated with the Kaplan-Meier method. The Cox proportional hazards model was used for multivariate analysis to determine the independent prognostic factors. Furthermore, we used Holm and Benjamini-Hochberg procedures to adjust for multiple comparisons in setting the level of significance. The median follow-up period was five years and two months (range, four to 195 months). RESULTS: The expression of interferon-α/ß receptor was positive in eighteen (45%) of the forty patients with high-grade osteosarcoma. American Joint Committee on Cancer surgical stage IIA, a good histologic response to chemotherapy, and expression of interferon-α/ß receptor correlated significantly with better disease-free survival (p < 0.05). Multivariate analysis showed that interferon-α/ß receptor expression alone retained its power to predict an improved prognosis (p = 0.042). There were no significant variables after corrections for multiple comparisons. CONCLUSIONS: Interferon-α/ß receptor may be a useful marker for assessing tumor prognosis in patients with osteosarcoma and may play an important role in tumor progression. These findings are encouraging and support the ongoing clinical trials of adjuvant interferon-α therapy by the multinational Osteosarcoma Study Group. Our pilot study was based on a small sample size, and larger trials are needed to confirm this finding. LEVEL OF EVIDENCE: Prognostic Level II. See Instructions to Authors for a complete description of levels of evidence.

Oncolytic vesicular stomatitis virus administered by isolated limb perfusion suppresses osteosarcoma growth.

A significant limitation to oncolytic virotherapy in vivo is the lack of a clinically relevant means of delivering the virus. We evaluated the oncolytic activity of vesicular stomatitis virus (VSV) in human osteosarcoma cells and explored isolated limb perfusion (ILP) as a novel oncolytic virus delivery system to extremity sarcoma in immune-competent rats. Human and rat osteosarcoma cells transduced with rVSV-lacZ uniformly expressed ß-gal. VSV was fully capable of replicating its RNA genome in all osteosarcoma cell lines, and efficiently killed them in time- and dose-dependent manners, whereas normal bone marrow stromal cells were refractory to the virus. VSV delivered by ILP inhibited growth of osteosarcoma xenografts more potently than that injected intravenously and intratumorally in the hind limb of immune-competent rats. Histopathological sections of tumor lesions treated by ILP-delivered VSV showed positive for VSV-G protein. There were no VSV-G expressions in perfused leg muscle, nonperfused leg muscle, brain, lung, and liver in VSV-treated rats. Our findings show efficient VSV gene expression and replication in osteosarcoma cells, suggesting that osteosarcoma may be a promising target for oncolytic virotherapy with VSV. Furthermore, we firstly showed that ILP of VSV against extremity sarcoma caused antitumor activity.

Alternative lengthening of telomeres as a prognostic factor in malignant fibrous histiocytomas of bone.

BACKGROUND: Malignant fibrous histiocytoma (MFH) of bone is a rare primary malignant neoplasm. Recent studies indicate a positive correlation between the telomere maintenance mechanism and tumour aggressiveness in sarcomas. This study was undertaken to analyse the clinical significance of telomere factors in primary tumour samples from patients with MFHs of bone. MATERIALS AND METHODS: Telomerase activity was measured in ten bone MFH specimens using a PCR-based TRAP assay. Telomere length was assessed using gel hybridisation. Quantitative detection of human telomerase reverse transcriptase (hTERT) was performed by real-time PCR. RESULTS: Telomerase activity and hTERT expression were detectable in 100% of tumour samples and 50% of tumour samples had evidence of engagement of the alternative lengthening of telomere (ALT) mechanisms. ALT was a significant prognostic risk factor (p = 0.0316). CONCLUSION: This study suggests that the presence of ALT telomere maintenance mechanisms indicates a poor prognosis for bone MFH patients.

Minodronate for the treatment of osteoporosis.

Minodronate, a new nitrogen-containing bisphosphonate, was developed in Japan. It was the first drug to demonstrate significant prevention of vertebral fractures in Japanese patients with osteoporosis in a phase III doubleblind comparative study. As a result of positive data from clinical trials in Japan minodronate was granted a Japanese marketing approval for the treatment of osteoporosis on January 21, 2009. In vitro studies demonstrated that minodronate is one of the most potent inhibitors of bone resorption among currently available bisphosphonates. Preclinical studies demonstrated the inhibitory effect of minodronate on the decrease in the bone mineral density (BMD) in ovariectomized rats, dogs and monkeys. Daily oral minodronate was safe, well tolerated and effective in reducing vertebral fracture risk in postmenopausal women with established osteoporosis. The effects on lumbar and hip BMD and the safety profile of minodronate are comparable to those of alendronate. These data suggest that minodronate is a promising new potent bisphosphonate for the treatment of osteoporosis.

Telomeres and telomerase in sarcomas.

Telomeres of human tumor cells have two types of telomere maintenance mechanisms by telomerase activation and alternative lengthening of telomeres (ALT). Although over 80% of all carcinomas rely on telomerase activity to maintain stable telomere length, many types of sarcoma elongate telomeres consistent with ALT in the absence of telomerase activity. Recently, the presence of telomerase activity and ALT in several sarcomas was examined extensively, and recent studies indicate a positive correlation between the telomere maintenance mechanism and tumor aggressiveness in several sarcoma types. We reviewed both the activation of telomere maintenance in a variety of common bone and soft tissue sarcoma subtypes, and the consequences of telomere maintenance mechanisms with respect to the clinical characteristics.