Pulmonary toxicity of tungsten trioxide nanoparticles in an inhalation study and an intratracheal instillation study.

OBJECTIVES: We conducted inhalation and intratracheal instillation studies in order to examine the effects of tungsten trioxide (WO3 ) nanoparticles on the lung, and evaluated whether or not the nanoparticles would cause persistent lung inflammation. METHODS: In the inhalation study, male 10-week-old Fischer 334 rats were classified into 3 groups. The control, low-dose, and high-dose groups inhaled clean air, 2, and 10 mg/m3 WO3 nanoparticles, respectively, for 6 h each day for 4 weeks. The rats were dissected at 3 days, 1 month, and 3 months after the inhalation, and the bronchoalveolar lavage fluid (BALF) and lung tissue were examined. In the intratracheal instillation study, male 12-week-old Fischer 334 rats were divided into 3 subgroups. The control, low-dose, and high-dose groups were intratracheally instilled 0.4 ml distilled water, 0.2, and 1.0 mg WO3 nanoparticles, respectively, dissolved in 0.4 ml distilled water. The rats were sacrificed at 3 days, 1 week, and 1 month after the intratracheal instillation, and the BALF and lung tissue were analyzed as in the inhalation study. RESULTS: The inhalation and instillation of WO3 nanoparticles caused transient increases in the number and rate of neutrophils, cytokine-induced neutrophil chemoattractant (CINC)-1, and CINC-2 in BALF, but no histopathological changes or upregulation of heme oxygenase (HO)-1 in the lung tissue. CONCLUSION: Our results suggest that WO3 nanoparticles have low toxicity to the lung. According to the results of the inhalation study, we also propose that the no observed adverse effect level (NOAEL) of WO3 nanoparticles is 2 mg/m3 .

Long-term observation of pulmonary toxicity of toner with external additives following a single intratracheal instillation in rats.

OBJECTIVES: Along with technological innovations for improving the efficiency of printing, nanoparticles have been added to the surface of toners, and there is concern about the harmful effects of those components. We investigated, through a long-term observation following intratracheal instillation using rats, whether exposure to a toner with external additives can cause tumorigenesis. METHODS: Female Wistar rats were intratracheally instilled with dispersed toner at low (1 mg/rat) and high (2 mg/rat) doses, and the rats were sacrificed at 24 months after exposure, after which we examined pulmonary inflammation, histopathological changes, and DNA damage in the lung. Rats that had deceased before 24 months were dissected at that time as well, to compare tumor development. RESULTS: Although alveolar macrophages with pigment deposition in the alveoli were observed in the 1 and 2 mg exposure groups, no significant lung inflammation/fibrosis or tumor was observed. Since immunostaining with 8-OHdG or γ-H2AX did not show a remarkable positive reaction, it is thought that toner did not cause severe DNA damage to lung tissue. CONCLUSION: These results suggest that toner with external additives may have low toxicity in the lung.

Dynamic evaluation of the spinal cord in patients with cervical spondylotic myelopathy using a kinematic magnetic resonance imaging technique.

STUDY DESIGN: This prospective study analyzed preoperative and postoperative dynamic changes of the spinal cord in patients with cervical spondylotic myelopathy. OBJECTIVE: (1) To study preoperative kinematic characteristics of the spinal cord in patients with cervical spondylotic myelopathy and compare results with healthy individuals, (2) to understand the dynamic changes of the spinal cord after posterior decompression, and (3) to discover whether the degree of posterior shifting is correlated with surgical outcomes using kinematic magnetic resonance imaging (MRI). SUMMARY OF BACKGROUND DATA: Laminoplasty for cervical myelopathy increases the space occupied by the spinal cord leading to a decompressive effect on the cord. However, no consecutive studies have reported the kinematic characteristics of the cervical spine in patients with cervical spondylotic myelopathy both preoperatively and postoperatively. Additionally, there have been no reports investigating the effects of posterior cord shifting in the neutral and maximum flexion and extension positions on surgical outcomes after cervical laminoplasty. METHODS: Twenty cervical spondylotic myelopathy patients who underwent extensive laminoplasty and 20 healthy individuals were examined. Preoperative and postoperative MRI records were available in all cases. The cervical spines of the subjects were examined in the neutral and maximum flexion and extension positions using an MRI scanner. Sagittal T1-weighted images were obtained at 12 different angles. Images were analyzed for the distance between the dorsal edge of the vertebral column and the center of the cord at each disc level using NIH image software. RESULTS: Average cord distances (L value) in the neutral position and maximum extension position at C4/5 was significantly smaller than those at the other disc levels. The spinal cords of the patients after laminoplasty moved dorsally in the enlarged spinal canal in the neutral position, and in the maximum flexion and extension position. However, the degree of posterior spinal cord shifting was not correlated with surgical outcomes. CONCLUSIONS: Cord distances are relatively smaller at C4/5 and C5/6 levels, resulting in a narrowing of the posterior subarachnoid space with posterior cord compression in patients with cervical spondylotic myelopathy. The outcome of surgery was not correlated with the magnitude of postoperative backward shifting of the spinal cord, although the spinal cord of patients after posterior decompression moved significantly dorsally at any of the flexed, neutral, or extended spinal positions. Thus numerous factors might affect the postoperative outcomes.

Biceps tendinitis caused by an osteochondroma in the bicipital groove: a rare cause of shoulder pain in a baseball player.

Tendinitis of the long head of the biceps brachii muscle is commonly seen in athletes who do repetitive overhead motions. Common causes of biceps tendinitis include impingement syndrome, subluxation of the biceps tendon, and attrition tendinitis, whereas biceps tendinitis secondary to a bone neoplasm is rare. A case of biceps tendinitis caused by an osteochondroma arising in the left humeral bicipital groove in a 25-year-old male baseball player is reported. The tumor was hook-shaped, originated from the inferomedial portion of the humeral lesser tubercle, and surrounded the biceps tendon. Symptoms of increasing pain and inability to throw resulted from direct irritation of the biceps tendon by the tumor. Total excision of the tumor relieved the symptoms within 3 weeks. To our knowledge, there have been no reported cases in the English-language literature of biceps tendinitis caused by an osteochondroma.

Establishment and characterization of a KIT-positive and stem cell factor-producing cell line, KTHOS, derived from human osteosarcoma.

Osteosarcoma is a malignant bone tumor that commonly affects adolescents and young adults. In the present study a human osteosarcoma cell line, KTHOS, was established from a primary osteosarcoma lesion in the distal femur of a 16-year-old girl. After 106 passages, the KTHOS cell line retained the biological characteristics of osteosarcoma. The KTHOS cells had spindle to pleomorphic cytoplasm with round to ovoid nuclei containing multiple prominent nucleoli, as expected based on the mesodermic origin of osteoblasts. The KTHOS cells were immunoreactive for osteocalcin, osteonectin, stem cell factor (SCF), and KIT (CD117). Reverse transcriptase-polymerase chain reaction indicated that the KTHOS cell line expressed mRNA for SCF and KIT. The KTHOS cells produced relatively high amounts of soluble SCF as determined by enzyme-linked immunosorbent assay. The results suggest that cell proliferation of the KTHOS cell line might be involved in autocrine and/or paracrine loops of the SCF/KIT signaling system. The KTHOS cell line is a novel human osteosarcoma cell line that releases SCF and expresses KIT. This cell line can be used for studies to explore the mechanisms for oncogenesis of human osteosarcomas.

Inhibitory effect of STI571 on cell proliferation of human malignant fibrous histiocytoma cell lines.

BACKGROUND: Malignant fibrous histiocytoma (MFH) is one of the most common high-grade sarcomas in bone and soft tissue and, due to its chemo-resistance, the prognosis of the disease is poor. ST1571 is a tyrosine kinase inhibitor that was initially developed as a BCR/ABL inhibitor for chronic myeloid leukemia patients. STI571 also selectively inhibits platelet-derived growth factor receptors (PDGFRs) and c-kit. We examined the expression of PDGFRs and c-kit in human MFH cell lines, and the effect of STI571 on cell proliferation. MATERIALS AND METHODS: Four human MFH cell lines (TNMY1, GBS-1, Nara-F and Nara-H) were used. mRNA expression of the receptor tyrosine kinases (PDGFRs and c-kit) was analyzed using reverse transcription-polymerase chain reaction, and the inhibitory effect of STI571 on cell proliferation was analyzed using the MTS assay technique. RESULTS: PDGFRalpha mRNA was expressed in TNMY1 and GBS-1, and PDGFRbeta and c-kit mRNAs were expressed in TNMY1, GBS-1 and Nara-F. All three of these mRNAs were absent in Nara-H. STI571 inhibited cell proliferation of TNMY1, GBS-1 and Nara-F in a dose- and time-dependent manner, but cell proliferation of Nara-H was not inhibited by ST1571 at concentrations of 10 microM or less. CONCLUSION: STI571 significantly inhibited proliferation of the three human MFH cell lines that expressed mRNAs of target receptor tyrosine kinases. The inhibitory effect of ST1571 on cell proliferation in these three cell lines might be due to decreased tyrosine kinase activity. STI571 might be a potent chemotherapeutic agent for human MFHs.

Expression of transforming growth factor beta isoforms and their receptors in malignant fibrous histiocytoma of soft tissues.

PURPOSE: Transforming growth factor beta (TGF-beta) is a multifunctional growth factor that variably affects proliferation, differentiation, and extracellular matrix formation. Little information is currently available on the TGF-beta expression in malignant fibrous histiocytoma (MFH). The aims of the present study were to investigate the expression of TGF-beta isoforms and their receptors in human MFH specimens. EXPERIMENTAL DESIGN: The expression of TGF isoforms, and TGF-beta receptors (TGF-beta R1 and -beta R2) were immunohistochemically evaluated in 43 paraffin-embedded MFH specimens. Furthermore, the correlation of the TGF-beta and receptor expression with tumor proliferative activity assessed by MIB-1 indices was analyzed. RESULTS: Positive immunoreactivity for TGF-beta1, -beta2, and -beta 3 was identified in tumor cells of 42, 40, and 38 of the 43 MFHs, respectively. In each TGF-beta isoform immunostaining, the specimens were divided into two groups based on the number of positive tumor cells: those with low (<25%) and those with high (>==25%) immunoreactivity. There were no statistically significant differences in the MIB-1 indices between the two groups. Positive immunoreactivity for TGF-beta R1 and -beta R2 was identified in tumor cells of 36 and 24 of the MFHs, respectively. The specimens were divided into two groups based on their receptor expression patterns: those with both TGF-beta R1- and -beta R2-positive immunoreactivity (n = 23), and those with both or either TGF-beta R1- and -beta R2-negative immunoreactivity (n = 20). The MIB-1 indices in the both-TGF-beta R1- and -beta R2-positive group were significantly higher than those in the other group (P = 0.0102). There was no significant difference in pulmonary metastasis ratios between the two groups. CONCLUSIONS: These findings strongly suggest an association of the TGF-beta ligand/receptor system with a significantly higher MIB-1 index in human MFHs. Investigation of the TGF-beta R1 and -beta R2 coexpression might be useful in predicting tumor behavior of MFHs.

Expression of betacellulin, heparin-binding epidermal growth factor and epiregulin in human malignant fibrous histiocytoma.

BACKGROUND: Heparin-binding epidermal growth factor (HB-EGF), betacellulin (BTC) and epiregulin (EPR) are members of the EGF system and involved in the cell growth of various epithelial malignancies. There have been no reports on the HB-EGF, BTC and EPR expression in mesenchymal malignancies of fibrohistiocytic origin including malignant fibrous histiocytoma (MFH). MATERIALS AND METHODS: We investigated the expression of HB-EGF, BTC, EPR and EGF-receptor (EGF-R) in 43 human MFH tissue samples using immunohistochemical techniques. RESULTS: Positive immuno-reactivity for HB-EGF, BTC, EPR and EGF-R was identified in 28 (65%), 7 (16%), 43 (100%) and 36 (84%) out of the 43 MFH cases analyzed, respectively. Coexpression of HB-EGF/BTC, BTC/EPR and HB-EGF/EPR was observed in 6 (14%), 7 (16%) and 28 (65%) of the MFHs, respectively. Coexpression of HB-EGF/EGF-R, BTC/EGF-R and EPR/EGF-R was observed in 25 (58%), 6 (14%) and 36 (84%) of the MFHs, respectively. CONCLUSION: These results revealed that HB-EGF, BTC and EPR are expressed not only by epithelial tumor cells, but also by MFH cells. It is suggested that HB-EGF and EPR might be more important tumor growth regulators of MFH through autocrine or paracrine pathways, when compared with BTC.

Coexpression of hepatocyte growth factor and its receptor c-Met correlates with high MIB-1 proliferative index in malignant fibrous histiocytoma.

Hepatocyte growth factor (HGF) is a multifunctional cytokine that variably affects cell motility, proliferation, and morphogenesis. Little information is currently available on the HGF and its receptor c-Met expression in malignant fibrous histiocytoma (MFH). We immunohistochemically investigated the HGF and c-Met expression in 43 MFH tissue specimens. Furthermore, the correlation of the HGF and c-Met expression with tumor proliferative activity assessed by MIB-1 indices was analyzed. Our results showed that positive cytoplasmic immunoreactivity for HGF and c-Met was identified in tumor cells in 36 (84%) and 20 (47%) of the 43 MFH cases analyzed, respectively. Coexpression of HGF and c-Met was observed in 20 (47%) of the 43 MFHs, and was correlated with high MIB-1 proliferative indices (p = 0.0446). These findings strongly indicate that the HGF/c-Met signaling system plays an important role in promoting cell proliferation of human MFHs via an autocrine loop.

Apophysitis of the ischial tuberosity mimicking a neoplasm on magnetic resonance imaging.

We present multimodality imaging features of an ischial tuberosity apophysitis in a 13-year-old boy who was an active baseball pitcher. Roentgenography of the pelvis and computed tomography showed mild irregularity in the inferior margin of the left ischial tuberosity. T1-weighted MRI showed a wide area with low signal intensity in the left ischial body; T2-weighted fat-suppression images showed areas with markedly high signal intensity in the ischial apophysis and body and the surrounding periosteum; contrast-enhanced T1-weighted fat-suppression MRI showed that the ischial body, surrounding periosteum, and origin of the hamstring muscles strongly enhanced; technetium-99m scintigraphic scans showed increased isotope uptake in the entire ischial body. Histological specimens obtained from the bone showed increased osteoblastic activity, edema, and proliferation of benign spindle cells and small vessels in the bone marrow spaces. In the present case, because MR imaging demonstrated extensive signal abnormalities involving the apophysis, periosteum, and intramedullary portion of bone, a neoplasm could not be excluded, and a biopsy was undertaken.

Amphiregulin and epidermal growth factor receptor expression in human malignant fibrous histiocytoma of soft tissues.

BACKGROUND: Amphiregulin is a member of the epidermal growth factor (EGF) system and a potent mitogen for various epithelial tissues. Little information, however, is currently available on the amphiregulin and EGF receptor (EGF-R) expression in mesenchymal malignancies of a fibrohistiocytic origin including malignant fibrous histiocytoma (MFH). MATERIALS AND METHODS: We investigated the amphiregulin and EGF-R expression in 43 human MFH tissues using immunohistochemical techniques. Furthermore, the correlation of the ligand and the receptor expression with tumor proliferative activity assessed by MIB-1 indices was analyzed. RESULTS: Positive immunoreactivity for amphiregulin and EGF-R was identified in 34 (79%) and 36 (84%) of the 43 MFH cases analyzed, respectively. Coexpression of amphiregulin and EGF-R was observed in 30 (70%) of the 43 MFHs analyzed. There were no significant differences in MIB-1 indices between both the amphiregulin and EGF-R-positive MFHs and the remaining MFHs. CONCLUSION: These results show that amphiregulin is expressed not only by epithelial tumor cells but also by MFH cells. Our data provide evidence indicating the presence of an autocrine mechanism of proliferation control involving the amphiregulin/EGF-R signaling system in human MFHs.

Neurilemmoma in the foot as a cause of heel pain: a report of two cases.

Two cases of deep-seated neurilemmoma that arose from plantar branches of the posterior tibial nerve and caused chronic heel pain are described. At the initial examination, one case was misdiagnosed as tarsal tunnel syndrome and the other was overlooked as plantar fasciitis; both cases were treated for long periods prior to operation. Deep-seated neurilemmomas in the foot can easily be overlooked and misdiagnosed as tarsal tunnel syndrome or plantar fasciitis because of the rarity, absence of palpable mass, and similarity of symptoms to those of other frequently encountered foot disorders. Magnetic resonance imaging provides the best modality for differential diagnosis. In the present cases, surgical excision of the tumors resulted in immediate and complete relief of chronic heel pain. Surgeons should consider neurilemmoma as a cause of persistent chronic heel pain despite the rarity of the disease.

Periosteal osteoblastoma of the distal femur.

Osteoblastomas located on the surface of the cortical bone, so-called periosteal osteoblastomas, are extremely rare. We report on a case of periosteal osteoblastoma arising from the posterior surface of the right distal femur in a 17-year-old man. Roentgenographic, computed tomographic, magnetic resonance imaging, and histologic features of the case are presented. Periosteal osteoblastoma should be radiologically and histologically differentiated from myositis ossificans, avulsive cortical irregularity syndrome, osteoid osteoma, parosteal osteosarcoma, periosteal osteosarcoma, and high-grade surface osteosarcoma. Although periosteal osteoblastoma is rare, this tumor should be included in the differential diagnosis of surface-type bone tumors.

Allograft replacement of common iliac artery mycotic aneurysm caused by Bacteroides fragilis vertebral spondylitis--a case report.

Mycotic aneurysm secondary to vertebral spondylitis is a rare but life-threatening pathology with high mortality and morbidity. The authors describe a successfully treated case of mycotic aneurysm of the common iliac artery complicated with vertebral spondylitis in a 74-year-old man. Under midline laparotomy, complete debridement of the infected tissues, in-situ replacement of the common iliac artery with cryopreserved aortic allograft, and iliac bone autotransplantation and omentopexy to fill the debrided cavity were performed. The postoperative course was uneventful, and he remains well 3 years after his operation without persistent infection or allograft rejection.

Expression of stem cell factor and lack of c-kit expression in malignant fibrous histiocytoma of soft tissues.

BACKGROUND: Little information is available on the expression of stem cell factor (SCF) and its receptor c-kit in soft tissue tumors of a fibrohistiocytic origin, including malignant fibrous histiocytoma (MFH). MATERIALS AND METHODS: We investigated the endogenous expression of SCF and c-kit in 43 MFH tissue samples using immunohistochemical techniques. Furthermore, we examined the correlation of SCF expression in MFHs with proliferative activity assessed by mitotic indices and MIB-1 immunohistochemical staining. RESULTS: Positive immunoreactivity for c-kit was identified in tumor cells of only one MFH case, while the remaining 42 cases were negative. In the one positive case, immunohistochemical staining was focal. Positive immunoreactivity for SCF was identified in 31 out of 43 cases studied (72%, focal; 11, moderate; 6, diffuse; 14). There were no significant differences in the MIB-1 and mitotic indices between the SCF-positive and negative groups. CONCLUSION: Our data indicate that any direct autocrine effects of the SCF/c-kit system on cell growth regulation are precluded in most MFH cases studied, but it is speculated that SCF might indirectly influence tumor growth by promoting local neovascularization.

Immunohistochemical analysis of platelet-derived growth factor and its receptors in soft tissue malignant fibrous histiocytoma.

BACKGROUND: Little information is available regarding the expression of platelet-derived growth factor (PDGF) isoforms and their receptors in soft tissue malignant fibrous histiocytoma (MFH). MATERIALS AND METHODS: We investigated expression of PDGF isoforms and their receptors (PDGF-R alpha and -R beta) in 43 MFH tissue specimens using immunohistochemical techniques. Furthermore, we examined the correlation of PDGF expression in MFHs with proliferative activity assessed by MIB-1 immunohistochemical staining. RESULTS: Positive cytoplasmic immunoreactivity for PDGF-AA, -BB and -AB was identified in tumor cells of 28 (66%), 4 (10%) and 26 (61%), respectively, of the 43 MFHs analyzed. Positive cytoplasmic immunoreactivity for PDGF-R alpha and -R beta was identified in tumor cells of 41 (95%) and 32 (74%), respectively, of the MFHs. Thirty-four (79%) MFHs coexpressed one or more PDGF isoforms and their corresponding receptors. In PDGF-AA immunostaining, MIB-indices in the high immunoreactivity group (> 10% of tumor cells) were significantly higher than those in the low immunoreactivity group (< 10% of tumor cells) (p = 0.031). CONCLUSION: Our data provide evidence to support the presence of an autocrine/paracrine mechanism of proliferation control involving the PDGF ligand/receptor system in human MFHs.

Extraabdominal desmoid tumor in a surgical scar of a patient with Sprengel's deformity.

Patients with an extraabdominal desmoid tumor have multiple minor bone abnormalities. The authors describe a rare case of an extraabdominal desmoid tumor that developed in a scar 2 years after surgery for Sprengel's deformity of the right shoulder in an 8-year-old girl. The association between the tumor and Sprengel's deformity has never been reported previously. Antecedent surgical trauma might play a role in the development of this tumor.

Expression of stem cell factor and c-kit in human malignant fibrous histiocytoma cell line (TNMY1).

BACKGROUND: The expression of c-kit and/or its ligand, stem cell factor (SCF), has been related to tumor proliferation, in several tumor systems. MATERIALS AND METHODS: We analyzed the expression of the SCF/its receptor (c-kit) mRNA and the production of soluble SCF in a human malignant fibrous histiocytoma (MFH) cell line (TNMY1). RESULTS: Immunocytochemical analysis revealed that the TNMY1 cells were positive for both SCF and c-kit. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed that the TNMY1 cell line expressed mRNA for SCF and c-kit. By using an enzyme-linked immunosorbent assay (ELISA), the TNMY1 cells were found to produce relatively high amounts of soluble SCF. However, the addition of soluble SCF to the TNMY1 cells did not alter the in vitro growth ability of the cells. CONCLUSION: Our data showed that the MFH cells produced consistent amounts of SCF but did not demonstrate autocrine growth modulation. Thus, SCF secretion may have a paracrine activity in the growth of MFH cells.